Horticulture

Although it may not be excluded, it seems unlikely that A. tortilis once grew on the site proper. This tree penetrates the coastal plain via the main wadi branches but is not found today in the near vicinity of Berenike. Both its salt non-tolerant status and its deep, penetrating root may prevent successful establishment on this limestone outcrop in a salt-marsh area. More to the south, however, a large population of A. tortilis is present in the coastal plain area. Such a population of trees would certainly have been more accessible than the more scattered specimens in the middle and upstream part of the wadis. Another possibility that might explain its presence at Berenike is that we are dealing with plant remains that may have partly entered the trash deposits by natural agents. The use of large, spiny branches for fencing off kitchen gardens to protect them against grazing animals, as has been observed in Marsa Alam, 140 km north of Berenike, probably offers the most plausible explanation. Such branches will have disintegrated in the course of time and will have supplemented the soil with their diagnostic parts.

A similar use has been described by Wood for dead branches of nabq ,vertical grow which are used to make thorn bush hedges in Yemen.One seed from a dump area belongs to the baobab. With its unusually thick trunk and slender branches, giving the impression that these above ground parts are out of proportion, this tree has a characteristic appearance. The baobab has its natural habitat in the dry woodland savanna south of the Sahara, where it is frequently associated with tamarind , a tree that has also been attested for Berenike by its seeds. Two baobab trees are also recorded from two different localities in north Yemen . Whether these are relics of a larger native population or have to be considered as introductions is not clear. The current distribution of the baobab in India is confined to the northern part, including the northwest coast along the Gulf of Cambay, where it grows in and around the old ports of Janjira, Chaul, and Surat . According to Burton-Page, the tree has to be considered as an early fifteenth century introduction. This rules out the possibility of an import from the Indian continent. On the other hand, the presence of a baobab seed in Berenike clearly demonstrates that it was traded in much earlier times and that an earlier introduction into India cannot be ruled out. If the seed unearthed from Berenike originates from the East African savanna, the most probable area of origin is northeast Sudan, from where it could have been offered for trade in Ptolemais Thêrôn, a port located at lat 18°40′ N and which was only accessible by small vessels .

As the baobab is almost not recorded from the Horn of Africa, a second possibility is that it originates from east tropical Africa, from where it could have been exported from Rhapta , according to the Periplus Maris Erythraei the only port of trade on the East African coast south of Opone. Because a return voyage from Berenike to Rhapta would take about a year and a half, its seems more likely that trade items from this area were subjected to distributive trade and offered for sale in the so-called “far side” ports in Somalia. This might also have been the case for the baobab. On the other hand, the uncertain status of baobab in north Yemen does not exclude the possibility that the baobab was imported from a far less distant place. The baobab is a highly valuable tree because all parts can be used in a variety of ways. The hard, indehiscent fruits of the baobab measure 15 to 20 cm in length and contain a great number of seeds imbedded in pulp. The seeds can be eaten fresh or roasted and contain up to 15 percent oil. The fruit pulp contains tartaric acid, which is also edible and can be mixed with water or milk. Additionally, both seeds and fruits have medicinal properties . Today, whole fruits, mostly sprayed with fancy colors, are offered for sale in fl orist shops and are used for making bouquets of dried flowers.Bulb remains of garlic have been evidenced from the Second Intermediate period until the Islamic period. In Berenike, a reasonable number of bulb bases and bulb scales from garlic were found in different trash dumps, giving the impression that garlic was cultivated locally and consumed in reasonably large quantities. Both garlic and onion are well represented in the archaeobotanical record of Egypt.

Within the spectrum of vegetables and herbs, both species belong to a small group of species that have a good chance of ending up in an archaeobotanical record. Edible parts from vegetables and herbs such as lettuce , thyme , and mint have a very small chance of being discovered, even in Egypt where preservation conditions are excellent, due to the extreme arid climate. If they are found at all, it is their seeds that are unearthed. This, in turn, is exceptional because these plants are normally harvested before seed setting. In fact, the few seeds that have been found can be considered as those meant for sowing or as seeds that originate from plants that have bolted, which makes more sense. Other vegetables and herbs, such as saffron, the orange-red stigmas of the crocus , are rare because they concern plant parts that cannot be propagated. When the edible parts of plants are also used for propagation, they are adapted to living in the soil, which favors a good preservation. This is the case with bulbs of garlic and onion and with the seeds and fruits of fennel , coriander , and dill . Almond is one of the fruits that has been imported to Berenike from the Mediterranean area. Although no large quantities were found, fragments are present in many samples and suggest that this fruit was available on a regular scale. In addition to the records from the Greco-Roman and Byzantine periods, almond fruits as well as walking-stick handles made from the almond tree are recorded from several archaeological contexts dated to the Eighteenth Dynasty . It is a point of discussion whether the almond was cultivated in Egypt during this period. Germer , for example, is of the opinion that the tree was once cultivated in Egypt because the small pot found in Tutankhamen’s tomb not only contained fragments of the endocarp , but also some whole fruits. Apricot has its origin in eastern Asia and was introduced in the Near East around the first century BC. From there it was introduced into the Mediterranean Basin where it became a well-established horticultural element . Archaeobotanical records of this perishable fruit are extremely rare. In fact, the only finds from within the Roman Empire are recorded from Egypt. One of them originated from the sacred animal complexes near Saqqara. The building of these cemeteries was started in the Nineteenth Dynasty and they remained in use in the Ptolemaic period. As is suggested by Germer , the kernel unearthed must have come from the Ptolemaic period. A second kernel is recorded by Thanheiser from Kellis in the Dakhla Oasis and is dated to the Roman period . In Berenike, two fragments of apricot were found in trash layers in trench 10. The most likely origin is the Mediterranean area,rolling grow table although it may also have been cultivated in northerly-located Fayum. The presence of a perishable fruit in such a remote corner of the Roman Empire clearly refl ects the luxury status of the food supply, though it may not have been available on a regular scale.In Egypt, mangrove vegetation consists predominantly of the Avicennia marina species and is restricted to the Red Sea coast, especially south of lat 25° N. This kind of vegetation is for the most part confined to the tropics and is indicative of muddy tidal waters. In the delta of Wadi Gimal, some 100 km north of Berenike, this plant is partly covered by sand hillocks, as a result of the silting of the shoreline zone. Apparently, A. marina can withstand the absence of nutritious mud to some extent. Along the Egyptian Red Sea this plant has mostly a shrubby habitus. It is possible that, in antiquity, the mangrove vegetation was more widespread. According to Strabo , mangrove vegetation was present along the whole coast of the Red Sea. The comparison of the mangrove tree with the olive tree and the laurel is, however, misinterpreted by Strabo and Pliny . Both writers are talking about olives when they describe the mangrove vegetation.

This exploitation of A. marina has had a long tradition. In the past, leaves were used as camel fodder during the summer, and shepherds ate the soft, green fruits . This kind of exploitation would certainly not have endangered this plant species. According to Mandaville the leaves are merely second choice because their salty taste. Additionally, Drar mentions of the use of the mangrove as fuel, resulting in the destruction of the mangrove vegetation, especially between Quseir and Wadi Gimal. Also Schweinfurth had already mentioned the use of A. marina as firewood, pointing to the high quality of the charcoal because it remains hot for a considerable period of time. Additionally, Schweinfurth reports the suitability of the tough and branching stems for constructing houses and fences. Today, this kind of exploitation is probably nonexistent. Ababda nomads still use branches of the twisted acacia in combination with mats or driftwood for making their shelters, although more and more concrete houses are becoming the norm. In the Gebel Elba area among Bisharin nomads, it was observed that wooden houses are built from a variety of woody species that grow in that particular environment, including arched branches of Cocculus pendulus. Furthermore, A. marina can also be used for tanning purposes. Exploitation of the tree for this purpose has resulted in the destruction of several mangrove forests . The presence of both leaves and wood of A. marina indicates that whole branches were harvested. Huge quantities of charcoal and some leaves of A. marina, intermixed with a minor fraction of Suaeda sp, were found in trench BE96-11, located in an industrial area southwest of the central town of Berenike . Most probably, this early Ptolemaic dump can be related to the cutting down of mangrove vegetation in the near vicinity of Berenike. Strabo, who visited Egypt just after the annexation of Egypt by the Romans, mentions that there were some convenient landing places for ships in Berenike . This indicates that at that time the bay was not silted up and, therefore, would have been too deep for the mangrove vegetation. Leaves used as fodder would have been a welcome by-product in the cutting down of the mangrove vegetation. It is also possible that mangrove wood was exported to Arabia, where it is still used for various kinds of construction, including houses, and as firewood. The mangrove was once the main cargo of specially constructed dhows that exported it from the East African coast . The only other archaeobotanical record of A. marina originates from Abu Sha’ar, a Roman settlement along the Red Sea coast 20 km north of Hurghada . Large quantities of leaves have been found here.Although some of its English names suggest membership of the palm family, the sugar date or desert date in fact belongs to the Balanitaceae, of which it is the only representative in Egypt. The shape of sugar dates is quite irregular and resembles that of the real date. But this resemblance does not hold for the anatomy of the fruit or the morphology of the seed. The inner layers of the fruit are woody and resistant to decay. The outer soft part of the fruit is not present anymore in sub-fossil specimens. The fruit pulp has a high sugar content of about 45 percent, while the relatively small seed is quite soft and yields 40 to 60 percent balanos oil. Balanos oil was used for the production of all kinds of perfumes. Like alcohol, fats and oils are capable of absorbing and retaining odors.

Only isolated specimens in inaccessible spots of rocky affluents succeed in flowering and fruit setting in such years. Other grasses remain small in size, even in favorable years, and it seems as if these grasses are present at the fringe of their distribution area. A good example is the presence of dwarf specimens installations, only one could be assigned to this variety. Several specimens were found between stones in a small affluent, which are relatively large in comparison with the variety typicum. The variety ehrenbergii Henr. is characterized by its scabrous lemmas, which are covered by thick hyaline-curved hairs. This variety is not mentioned for Egypt. According to Henrard , its distribution area stretches from Ethiopia and Eritrea to Arabia and Iran. In the more southerly Gebel Elba area, which is characterized by the presence of the Sudanese flora, in addition to the variety typicum, only two other varieties have been recorded, viz. pumila and aethiopica . Thus the presence of the variety ehrenbergii in Shenshef clearly points to a connection with more-southerly-located areas. One may wonder if the presence of this plant in the vicinity of Berenike can be related to the former trade contacts with Arabia, Ethiopia, and Eritrea.

The investigated localities in the Eastern Desert have many species in common, especially when they are connected by the same wadi system. In Khesm Umm Kabu and Qariya Mustafa ‘Amr Gama,plastic seedling pots which are located about 115 km apart, mutual species are rare, namely, Tamarix aphylla, Salvadora persica, and Trichodesma africanum. The number of unique species for a particular site is relatively small. Examples are Ifl oga spicata and Rumex simpliciflorus near Khesm Umm Kabu in the upstream part of Wadi Gimal. Obviously, both species were also found in the delta of this wadi and this shows that the dispersal within a specific wadi is not problematic.The high percentage of species that is present in more than one locality seems to indicate that seed dispersal, which is mainly dependent on wind and animals, is quite effective. Additionally, strong water currents after heavy rainfall transport a lot of sediment including the existing seed bank. Dispersal by the agency of water is, however, always restricted to the specific catchment area of a wadi system. Moreover, seeds are transported in either the direction of the Nile Valley or the Red Sea coast. The effectiveness of this kind of dispersal can be observed, for example, in drift litter along the Red Sea coast. Fruits of Zilla spinosa, Balanites aegyptiaca, and Neurada procumbens, for example, were frequently found in such concentrated organic debris.

Due to wind and animals, seeds succeed in reaching new wadi systems. Probably, the fl at coastal plain plays an important role in this respect as seeds that arrive on this coastal strip can easily be transported in a northerly or southerly direction, depending on the prevailing wind direction. Wadi branches in the coastal plain proper constitute no serious obstacles in this respect. I once witnessed a huge Zilla spinosa of almost 1.5 m in diameter rolling over the coastal plain. The wind was strong enough to lift this plant over a steep slope of a wadi branch of about 3 m high. The mountain slopes, however, constitute insurmountable barriers, resulting in large concentrations of tumbleweeds on specific spots . Dispersal of seeds by means of tumbleweeds, which lose their seeds during rolling, is an effective method in arid and semi-arid environments. Due to the scarcity of water, no closed vegetation is developed, and most plants can reach their optimal shape. This enables plants such as Zilla spinosa to develop into a spherical shape. The root of this species is twisted alternately to the left and right and weakens at the ground level after seeds are ripe. As a result, the plant is easily uprooted and becomes a tumbleweed. Not only spherical-shaped plants behave as tumbleweeds. For example, the procumbent Aizoon canariense also is uprooted and becomes a tumbleweed, even with part of the root still attached. The small seeds can easily escape from the cracks in the capsules. Finally, the whole plant becomes disintegrated. A special case in this respect seems to be Aristida funiculata, which might be described as “tumbleseeds.” Unlike the other two Egyptian species of this genus, namely, A. adscensionis and A. mutabilis, both column and awns of A. funiculata are long and stiff.

The total length of a spikelet is almost 8 cm. It has been observed several times that seeds form a cluster in such a way that the awns are interwoven and the seeds are projected on the outside of the clump. Another advantage of this species in this respect is that the articulation point is just below the base of the column. Due to this position, the shape of the clump of seeds is not affected when seeds break off during tumbling. Two such seed tumbles were found entangled in Convolvulus hystrix, showing that only part of the seeds were lost during their probably short trip. Dispersal by animals is possible by external carriage and by secretion via the guts. Spiny fruits, such as those of Arnebia hispidissima, or sticky ones, such as those of Boerhavia repens, are primarily transported via the outside of animals. The seeds of Acacia tortilis may serve as an example of the second mode of dispersal. Both unripe and ripe fruits of A. tortilis are partly found beneath the foliage, partly covered by sand. Such seeds need a secondary dispersal agent to find a safe site to germinate. More effective is the dispersal by camels internally. Camels have no problems with the long, stiff spines and are able to browse from the branches . Whereas twigs, leaves, and fruits are digested, the extremely hard seeds are excreted with the dung. Frequently, seedlings can be observed that germinate from inside camel dung . Owing to the wind, dung particles are quickly covered with sand, which in turn improves the growing conditions for the young tree. The number of seedlings that develop in a dung particle may vary considerably and will depend on the number of fruits excreted in a dung particle, as well as the number of mature, unaffected seeds in those fruits. Failed fertilization, abortion of seeds, and the attack of insects all contribute in reducing the number of viable seeds in a fruit . A concentration of 17 seedlings of A. tortilis proved to have originated from camel dung containing 18 seeds and covered by 10 cm of sand. Only when such camel dung is dropped at the edge of a wadi can one of the seedlings become a full-grown tree because strong water currents periodically destroy the vegetation in the central part of a wadi. For the same reason, these trees have a better chance of surviving along convex banks. One of the few shrubs that has frequently been observed in the central part of the wadi and is capable of surviving flooding is Chrozophora tinctoria, though its appearance is little more than that of a heavily damaged dwarf specimen.A variety of trees and shrubs were found in the vicinity of the Roman installations: Lycium shawii, Tamarix nilotica, T. aphylla, Suaeda monoica, Acacia tortilis, Salvadora persica, Balanites aegyptiaca, Calotropis procera, and Leptadenia pyrotechnica. The most common tree proved to be A. tortilis, found along the edges of many wadis, both in the mountains and in the coastal plain. Also around Berenike, this tree is frequently present in the small wadi branches. More substantial populations of acacias have been observed in the coastal plain south of Berenike. Despite its predominance, Acacia tortilis may be absent locally, such as in Wadi Gimal near Khesm Umm Kabu. Here, Balanites aegyptiaca and Calotropis procera are the predominating trees. Ababda nomads confirmed that Wadi Gimal, in particular, was characterized by B. aegyptiaca. In wadi branches dissecting the coastal plain,container size for raspberries this tree is a rather strange phenomenon. This is not the case with C. procera, which can frequently be observed in the coastal plain, especially south of Berenike. Tamarix nilotica and Suaeda monoica are halophytes and occur in the near vicinity of Berenike. On the higher levels of the salt marsh north of Berenike, several large hillocks of T. nilotica are currently present.

When several rows of such hillocks are present, the population functions as a natural barrier against penetrating windblown sand. In the salt marsh south of Berenike, T. nilotica is intermixed with S. monoica. Here, T. nilotica does not form hillocks and is present on the moreelevated levels, whereas S. monoica is concentrated in the lower levels of the salt marsh, together with the halophytic Halopeplis perfoliata. On both sides of the Red Sea coast, S. monoica gradually replaces the halophyte Nitraria retusa in a southerly direction. Halfway between Wadi Gimal and Ras Banas there is a transitional zone, whereas south of Ras Banas only populations of S. monoica are present . The Suaeda population south of Berenike dropped most of its leaves after the heavy showers in November 1996. The distribution of the salt-requiring S. monoica is confined to locations with a specific chloride content and the water table . Laboratory experiments with artificial solutions containing only traces of sodium resulted in the yellowing of leaves of S. monoica, as well as curling and wilting, whereas field observations revealed that seasonal fl uctuations have apparent effects on the distribution of this species . Other salt-tolerant species present in the salt marsh around Berenike are Cyperus conglomeratus, Zygophyllum album, and Z. simplex and an herbaceous Suaeda sp. Zygophyllum album has a wide ecological range with respect to the salt content of the soil, and Z. simplex is even less demanding as it also grows in the mountains, far away from the influence of the sea. The ecological preference of Z. coccineum, which is also rather common in the surroundings of Berenike, is less clear. The species has a large tolerance with respect to salinity and it is said to be indicative of limestone . In fact, both environmental conditions are present around Berenike, and each condition could explain its presence in itself. With respect to the salt content, possibly two ecotypes exist: a salt-tolerant and a nonsalt-tolerant one. It could be that the specimens growing in the vicinity of Berenike concern the salt-tolerant ecotype. Obviously, these specimens are full sized, whereas those found in the vicinity of the Roman settlements in the Red Sea mountains were less frequent and also of a reduced size. The population of Z. coccineum near Hitan Rayan might be of the nonsalttolerant type as this inland area does not witness the maritime influence of the Red Sea. Furthermore, the area is of Precambrian crystalline rocks and no limestone is present in Hitan Rayan or any of the surrounding hills . Salt-tolerant plant species are present near the shoreline on the supratidal flat , which consists of sandy, silty, and clay-like sediments and on a broad strip of the coastal plain adjacent to the sabkha. The latter is characterized by a salty horizon near the surface, which is quite difficult to penetrate. Mangrove vegetation is absent in the near vicinity of Berenike; its nearest population is located several kilometers north of Ras Banas. In Egypt this kind of vegetation mainly consists of Avicennia marina. South of lat 23° N it is also mixed with Rhizophora mucronata . Mangrove is present in calm, shallow water and on a fl at, muddy soil, permanently flooded or during high tides. Both protected lagoons and bays are suitable habitats. The northeast boundary of the present distribution is marked by some scattered specimens about 20 km north of Hurghada and becomes quite dense south of lat 25° N. At some localities this species also grows on sandy hills along the coast.Living in a desert environment essentially leads to its exploitation. Due to the limited carrying capacity of the environment, which is basically determined by the availability of water, people mostly live in relatively small groups that have adopted a nomadic life style. As a consequence, the material culture is strongly related to the bare essentials of life. The current arid climate of Egypt was established during the early second part of the Holocene and has had an intrinsic influence on the desert culture.

The series editors would like to thank the following organizations and individuals for enabling the eight years of archaeological work at Berenike on which the annual reports and final compilations such as the present volume are based: National Geographic Society, Netherlands Foundation of Scientifi c Research , Gratama Foundation, donors of the Berenike Foundation, Mallinson Architects of London, the University of Delaware, the American Philosophical Society, the Dorot Foundation, the Samuel H. Kress Foundation, Lotus Hotel in Cairo, Utopa Foundation, Organon Egypt, Eastmar Travel in Cairo, Philips Egypt, 3-Com Computer, Dionysus Systems, and in alphabetical order the following private donors: Carmine Balascio, Millie Cassidy, Bruce Gould, James Harrell, Charles Herndon, Susan Hodge, Dorothy Johnson II, Norma Kershaw, Carol Maltenfort, W. Weissman and W. Whelan.For a long time Berenike has been on the list of desiderata of many archaeologists. It was not until 1818, however, that G. B. Belzoni,raspberries in pots a weightlifter engaged in hunting antiquities in Egypt, identified Berenike south of the large Ras Banas peninsula.

Several visitors became interested in the site, and the expeditions that followed were mainly focused on excavating the temple dedicated to the Greek-Egyptian god Serapis, situated on the highest ground at Berenike . These visits, however, were short-lived due to the extreme desert conditions and related logistic problems. Food and water were not locally available and could be only delivered over large distances. In addition to the logistic problems, security reasons were also a serious hindrance to the realization of a large-scale excavation. The strategic location of the Greek-Roman harbor of Berenike has its modern counterpart in the presence of a large military base on Ras Banas and scattered bunkers on the limestone outcrop on which Berenike was founded. Furthermore, both Egypt and Sudan have a claim to the Hailab area, a triangular piece of land bordered along the Red Sea by Shelateen and Hailab. For these reasons, the Eastern Desert south of Marsa Alam was until recently only accessible with special permission. A couple of years after the Gulf War , it became possible to start the Berenike project, which included excavations at Berenike and Shenshef and the survey of the southern part of the Egyptian Eastern Desert. The excavations and surveys were directed by Prof. Dr. S. E. Sidebotham from Delaware University and Dr. W. Z. Wendrich from Leiden University.

After a pilot excavation season in 1994, large-scale excavations began and continued until December 2003, when the threat of an invasion of Iraq was at hand. Including the pilot study, a total of eight excavations seasons were conducted. The aim of the research project was to study the organization of trade and the subsistence economy at Berenike. Berenike was, together with Myos Hormos, an important harbor along the Red Sea coast. It was founded approximately 275 BC for the import of all kinds of luxury commodities from Africa south of the Sahara, Arabia, and India. Only at the beginning of the seventh century AD before the Arabs arrived in Egypt, was Berenike abandoned by the Romans. Several written historical sources deal with this trade, and these documents have been used for a long time to reconstruct this foreign trade. It remained uncertain, however, how reliable this picture was. Written sources are certainly not complete in their enumerations, and the interpretation of certain exotic commodities is still under discussion, as can be judged by the different translations of the Greek and Latin texts. The available information on the ancient trade contrasts sharply with that dealing with the subsistence economy of Berenike. Although located along the Red Sea, Berenike faced a desert climate that must have drawn heavily on its food supply. Much progress was therefore to be expected from the Berenike project, which had the cooperation of all kinds of specialists. This book synthesizes the results of the archaeobotanical research carried out over the years at Berenike and Shenshef and includes information that has been published in previous interim reports. The archaeobotanical research is based on the study of plant remains that have been preserved in both sites.

In Egypt, plant remains are predominately preserved by desiccation owing to the arid climate. Additionally, plant remains may become charred as a result of deliberate burning, such as in the offering of food items, or by accidental fire. The desiccated plant remains, in particular, are still in an excellent condition and facilitate, in most cases, identification to the level of species. In this way, a detailed list of plant species could be produced from Berenike and Shenshef, providing a solid basis for a reconstruction of the international trade of plant products and the food economy. The sub-fossil plant remains from Berenike originate from 794 samples and those from Shenshef from 86 samples. The samples from Berenike represent 34 different trenches, which are partly related with buildings and partly with trash deposits. Those from Shenshef originate from 10 different middens. Basically, each soil unit from a particular trench has been dry-sieved over a sieve with a mesh size of 4.0 mm. All sieve residues have been sorted out and have yielded many so-called “hand-picked” plant remains. A disadvantage of this procedure is that it is highly biased in favor of plants that produce large fragments. Cereals, for example, will only be secured in this way by the long rachis fragments in such sieve residues, whereas most of the grain kernels, threshing remains, and accompanying seeds of weed plants will get lost. To obtain a representative record of plant remains from Berenike and Shenshef, botanical samples were collected that have been processed by using sieves with a standardized mesh size of 5.0 mm, 2.0 mm, 1.0 mm, and 0.5 mm. The botanical samples from trash deposits, in particular, proved to be rich in well-preserved plant remains. Samples that were secured from the inside of buildings, on the other hand, were in most cases soil samples with only a low concentration of plant remains. Because these sediments were, for the greater part, considered as wind-blown secondary fillings of buildings, the plant records from these samples could not be used for an interpretation of the use of these buildings. For the reconstruction of agricultural practices, it is desirable to have at one’s disposal a considerable number of samples that presents one particular crop and associated weed plants. Unfortunately, almost all samples did contain at least two crops, such as wheat, barley, or lentil. This makes it frustrating to reconstruct the specific agricultural practices and also hampers the tracing of the origin of the crops. Most crops have acquired a broad tolerance to environmental factors as a result of selection processes, which is, for example, expressed by the many land races that have evolved. In most cases it is not possible to identify the area of origin of a cultivated plant found outside its production area, such as in Berenike, by studying the morphological features of its remains. Weed plants, on the other hand, are partly indicative of specific environmental conditions. Such plants can be used to trace the source of a particular crop plant indeed,blueberries in containers growing though its association with a particular crop should be evidenced by the archaeobotanical record. Samples with only a few plant species, rather than those with a high diversity, are suitable in this respect because one can usually unequivocally match these crops with their associated weed plants. Because the samples from Berenike and Shenshef did not meet this condition and also because of the large number of samples, it has been decided to categorize all plant records in several tables. Although Berenike was inhabited throughout the Ptolemaic and Roman periods, the archaeobotanical results are confined to the Roman period. It appeared that most of the Ptolemaic remains are located eastward of the Roman settlement. The few Ptolemaic contexts that have been unearthed had suffered seriously from salt seepage and did not yield identifiable plant remains. It was realized at an early stage in the project that the interpretation of the archaeobotanical records would benefit from the study of the current natural vegetation around the Roman installations and from observations of the local Ababda nomads who live in the southern part of the Eastern Desert.

For that reason, inventories have been made of the desert vegetation around the Roman installations during the various excavation seasons. Additionally, a separate visit was organized to the Gebel Elba area, located some 260 km south of Berenike, which was one of the source areas of some of the trade products. The ethnoarchaeobotanical research of the Ababda nomads was focused on the basic necessities of desert life with special emphasis on their material culture, their exploitation of the natural environment, and the possibilities of local food production. The reconstruction of the former natural vegetation is based on the archaeobotanical records, evidence from written sources, and the current vegetation. The interpretation of the cultivated plants found at Berenike and Shenshef summarizes their possible use, including the possibility of local food production, the differences in habitation periods, and the identification of their possible source of supply. The identification of the possible area of supply proved to be rather problematic because of the botanical richness of most of the samples. Determining the supply area is based on the natural distribution of the plant species, its archaeobotanical record from Egypt and the surrounding areas, and on the evidence from written historical sources, if available. A special topic related to the long-distance transport and the hot climate in Egypt is the preservation of food and the luxury status of food items. Based on the different methods of food preservation known from historical sources and those practiced today, the possible ways in which the various plant products might have been treated are discussed. Finally, the archaeobotanical record of exotic plant species from ancient Egypt is discussed, with special emphasis on those evidenced from Myos Hormos, the other important harbor along the Red Sea.Knowledge of Indian trade comes from various historical sources. Two of these sources, the Alexandrian Tariff and the Periplus Maris Erythraei, are of particular interest since they include a considerable list of botanical commodities that might have been traded through Berenike . Both sources provide first hand information. It is assumed that the author of the Periplus was an Egyptian Greek who had traveled at least along the African coast south to Rhapta and to the west coast of India. Contrary to other periploi that have survived, this Periplus is exceptional in its emphasis on trade items, in addition to information on the trade route proper. The Alexandrian Tariff was issued between AD 176 and 180 by Marcus Aurelius and enumerates 54 items subject to import duty at Alexandria on their way to Rome. It includes 20 different plant products, only half of which are also mentioned in the Periplus: costus, cassia, aloe, lykion, myrrh, malabathron, black and long pepper, nard, and aromatics. The Periplus, dated between AD 40 and 70, describes in great detail the trade routes from Myos Hormos, now identified as Quseir al-Qadim, and Berenike to India, including many harbors along the African, Arabian, and Indian coasts. The Erythraean Sea formerly included the Red Sea, the Gulf of Aden, and the Indian Ocean. The Periplus enumerates a vast number of import and export commodities, including 34 products of botanical origin, of which 18 are reported as import items from Berenike. The trade items mentioned in both documents represent a wide variety of plant parts: root; wood; bark; plant secretions such as resins, gums, and oils; leaves; flowers; seeds; and fruits, as well as whole plants . Plant secretions and seeds or fruits are the best-represented categories. A few trade items are either unspecified or not documented in such a way that a reliable identification can be made . The arrangement according to plant part is in some cases arbitrary. Aloe, for example, may be either a resin or fragrant wood, and both rhizomes and leaves were traded asnard. In some cases, trade items are reduced to a common trade name, such as “cassia” for “cassia turiana” and “xylocassia.” The scientific plant names are mainly based on Warmington , Miller , and Casson and have been updated where necessary.

Data were analyzed using a 3-way ANOVA with density of seed , application of B. bassiana , and date as factors . The commercial blueberry test site selected was located north of Bakersfield in Delano, CA. The trial began in August of 2008 and was conducted post blueberry harvest. The V. corymbosum varieties contained within the test area were, ‘Santa Fe’, ‘Jewel’, and ‘Star’. The most susceptible variety of blueberry to citrus thrips damage grown at the test site was the ‘Star’ variety and ‘Star’ was used consistently for evaluation of thrips numbers for all aspects of the trial . Our cooperator was interested in alternatives to traditional pesticides as the farm regularly was dealing with extremely high citrus thrips populations. For example, in 2008 the grower sprayed 5-10 times per field , rotating with traditional chemicals to reduce thrips impact on the subsequent year’s fruit set. Irrigation in all fields took place via drip irrigation with one water delivery emitter per line at each plant base , but additionally, one portion of the blueberry field was equipped with 360° overhead sprinklers. This irrigation setup provided the ideal situation to test B. bassiana under two watering regimes. The commercially available GHA strain is formulated to be mixed with water and for application via chemigation or as a foliar spray.

The label states that no surfactant is needed to keep the spores in suspension. However,plastic plants pots agitation alone in the 1,892.7 L holding tank was not sufficient to keep the material from precipitating, therefore 312.3 ml of Silwet L-77 was added to the tank mix. Mycotrol O® was applied directly to the soil surface with a gas-powered sprayer with a hand spray gun equipped with an adjustable flow meter. The dimensions of the plots were used to calculate the amount of material needed for both B. bassiana formulations . Plants in the test field were spaced every 0.92 m down each row, 3.35 m between each row, and each row was about 165 meters in length. Our studies were conducted inan 18-row section of a 4.04 ha field. The overhead sprinklers were spaced every 7 meters in the row and were located every other row for 12 rows. We chose to investigate the effectiveness of the B. bassiana colonized millet seed versus a Mycotrol O® soil application under two watering regimes, drip-line alone versus drip-line with overhead sprinkler, because B. bassiana conidia are highly subject to desiccation. Comparing the soil drench in both irrigation types with the colonized millet elucidated the effectiveness of the treatments when compared to the control. The blocks were laid out in a 3 x 2 factorial design, with each block consisting of most of five rows of blueberries , each being 27.4 m long . The berm used to grow blueberries at the commercial farm was 1.21 meters wide and each plot was 27.4 meters long. The spacing between adjacent rows was 3.35 m, while the spacing between the plants down a row was approximately 0.92 m with 30 plants per treatment plot . These dimensions result in 0.157 ha treated with raw spores but because the top of the berm was where thrips activity was evident and would be sampled, only 36% of the soil surface area was treated.

The Mycotrol O® label states that the maximum field rate is 6.9 L/ha mixed in 935.3 L/ha water. We therefore chose to apply the entire 6.9 L of Mycotrol O® in 378.5 L of water per ha directly to the berm with no application between the rows, which resulted in 100% of the per ha rate of product being applied to 36% of the area and allowed the maximum amount of active ingredient to be applied to the area that would have almost all thrips activity . Our field trial was intended to determine the extent to which B. bassiana might fit into a program projected to both control citrus thrips effectively and provide rotation among available chemistries so as to reduce thrips resistance evolution. Thus, we felt it was important to operate under the best possible conditions for thrips infection by Mycotrol O® , regardless of financial considerations, i.e. application of product at the maximum label rate in the area where thrips were most likely to be active. The amount of millet seed used in the field trail was calculated based on the area of the berm to be treated and likewise with the Mycotrol O® treatment, only 36% of the total field area was treated. The amount of seed used was one colonized seed/ 2 cm2 over an area of 576 m2 ; the fact that 0.45 kg of seed was needed per 840 cm2 resulted in the application of 3.40 kg of colonized millet seed for the 8 treated plots . Every other plant within the middle ten plants of the middle row of each plot were sampled with pupation emergence cages . These cages were placed tight against the base of each set of canes on the east side . With 5 cages per block and 4 replicate blocks per treatment, a total of 20 cages sampled thrips pupation per treatment over two sample periods, i.e. for two consecutive 3-day periods after the Mycotrol O® soil drench. The treatments were: no B. bassiana with and without overhead sprinkler; colonized millet seed with and without overhead sprinkler; and a soil drench of Mycotrol O® with and without overhead sprinkler . In total, data were collected from 240 emergence cages over the duration of the trial .

The colonized millet seed was set to imbibe water and allowed to sporulate for three days before application and was applied using a hand fertilizer applicator . Four days post application of the millet seed, the soil drench of Mycotrol O® was applied and pupation emergence cages were placed in the field and left out for 3 days . After three days, the sticky cards from each emergence cage were collected and replaced with new cards and the traps were switched to the next plant on the east side. These traps were left in the field to sample thrips for another 3 days . Because the traps were placed out every other plant, this ensured that all of the middle ten plants were sampled over the two, 3-day sampling periods . For two weeks before through two weeks after the applications of B. bassiana , counts were taken of thrips levels on plants twice per week. Beat samples were taken by beating random canes of flush foliage such that the thrips would fall onto a 12 x 12 cm black acrylic beat tray. The numbers of thrips on the beat trays were counted quickly in the field. The counts were taken twice per week from each of the 10 central “data plants” from the ‘Star’ variety of each of the 24 test plots. The new green flush growth was measured on three dates to record the amount of growth since the beginning of the fungal treatment applications to determine if there were differences based on the treatments and amount of water applied to the different plots. Measurements were made of the average cm of new shoot growth over the 6-week trial period. Due to the complex nature of the experimental design, i.e. treatments nested in a 5-way ANOVA , beat count data were analyzed using PROC MIXED and means were separated using Tukey’s test . Figure 3-2 shows the location of late second instar citrus thrips at death in the greenhouse study as well as those that located pupal refuges on the plant. Based on where they dropped off the plant,blueberry pot data indicated that more than 92% of the thrips would have pupated off the plant, likely in the soil near the base of the plant. Numbers did not vary significantly by location over the seven sample dates of this study; therefore data were pooled . A key result was that the proportion of second instar thrips crawling down the base of the plant was higher than the proportion dropping off the plant at distances measured past the base the plant . The four emergence cages placed under the field blueberry plants in each cardinal direction provided a means of sampling late second instar thrips moving towards the soil to pupae versus adults emerging out of the soil following pupation . Total numbers of thrips collected were pooled for the four traps in each direction at each respective location to determine which cardinal direction showed the most activity, and therefore was the most appropriate location to sample for citrus thrips in the field trial. Emergence cage data were summarized in two ways; the number of thrips moving off the plant to pupate in the leaf litter and the number of thrips emerging from the leaf . Data from the nested ANOVA generated p-values for direction , as well as distance grouping from the base of the plant . The cage closest to the base of the plant had significantly higher numbers of thrips emerging from the soil . Numbers of thrips trapped from the eastern cardinal direction were significantly higher for both mean numbers of thrips moving to and from the soil , indicating that for the field trial, emergence cages should be placed directly next to the base of the plant on the eastern side to sample the location that would have the most thrips activity.

Of the proportion of thrips not finding pupal refuge on the plants with colonized seed , 100% infection was seen with each of the different quantities of seed, i.e. each of 0.5, 1, or 2 seeds/cm2 was a sufficient density to infect and kill all late second instar thrips in the greenhouse study. No thrips were infected in the control treatment. There were insufficient data to conduct a 3-way ANOVA because all recovered thrips were infected with the fungus. Because all three densities tested were effective, we chose to utilize the most economical density in the field trial, i.e. 0.5 seeds/cm2 . In the split-plot design model, the whole plot factor was water and the split-plot factor was fungus treatment in a type three analysis of variance . Water, time and treatment were the main effects in the full model. Thrips levels measured on pupation traps at 3 days after treatment were lowest with colonized millet seed, intermediate with Mycotrol O ® , and highest in the untreated control . Additionally, there was fewer thrips counted in the colonized millet seed treatment than in Mycotrol O® treated plots . However, at time two , thrips levels with Mycotrol O® were no longer significantly reduced in relation to the control . While thrips levels measured using pupation traps were significantly less than observed in the control in all plots , thrips levels on plants measured using beat samples did not show a significant decrease , although comparing data with no overhead water that with overhead water, there appeared to be fewer numbers of thrips in the overhead sprinkler plots . The measurements from the new green flush growth in the overhead sprinkler treatments showed that those plants had longer growth than those without overhead sprinklers , but thrips numbers were not significantly lower on those plants. At none of the times when foliar beat counts were taken were there significant differences in thrips numbers across any of the three treatments . The ultimate goal of this work was to determine if the GHA strain of Beauveria bassiana could be used effectively as an alternative to traditional insecticides in commercial blueberries in California. Laboratory and greenhouse trials with Beauveria bassiana have shown variable success in controlling thrips and several other insect species , whereas field trials have shown limited overall success, but very few field trials included Thysanoptera . This is mainly due to the fact that climatic conditions in the laboratory and greenhouse situations are stable and often more humid than the ambient field environment in arid areas like most of California.

The trial was initiated in late fall 2000 in a drip-irrigated vineyard near Greenfield, Calif., and continued through the 2005 harvest. The vineyard was established in 1996 with Vitis vinifera L. cv. Chardonnay on Teleki 5C rootstock. Vine spacing was 8 feet between rows and 6 feet within rows. Annual rainfall normally ranges from 4 to 10 inches. Soil is elder loam with gravelly substratum. The vineyard was drip-irrigated from April to October.Row weed control treatments were: cultivation, post-emergence weed control only and pre-emergence herbicide , followed by post-emergence herbicide applications . Cultivations and herbicide applications were timed according to grower practices and label rates. Cultivations were carried out every 4 to 6 weeks during the growing season using a Radius Weeder cultivator . The cultivator used a metal knife that ran 2 to 6 inches below the soil surface cutting weeds off in the vine row; it had a sensor that caused it to swing around vines. Pre-emergence herbicides were applied in winter with a standard weed sprayer, and post emergence herbicides were applied in spring through fall as needed with a Patchen Weedseeker light-activated sprayer . An early and late-maturing cereal were chosen for the cover-crop treatments; legumes were not considered due to aggravated gopher and weed problems.

Cover-crop treatments in the middles were: no cover crop ,plant pot with drainage earlier maturing ‘Merced’ rye and later maturing ‘Trios 102’ triticale . Cover crops were planted with a vineyard seed drill in a 32-inch-wide strip in the middle of 8-foot-wide rows just before the start of the rainy season in November 2000 to 2004 . They were mowed in spring to protect vines from frost, and both cover-crop species senesced by summer. Prior to planting cover crops each November, row middles were disked to incorporate the previous year’s cover crop and stubble and prepare a seedbed. Weed control and cover-crop treatments were arranged in a 3 x 3 split block design with three replicate blocks covering a total of 23 vineyard rows . Each block contained six vine rows and six adjacent middles. Weed control treatments were applied along the entire length of each vine row ; cover-crop treatments were established along one-third of each middle and were continuous across the main plot treatments in each block. Each replicate main plot-by-subplot treatment combination included 100 vines. Soil compaction. Soil compaction was measured in the vine row in November or December 2003, 2004 and 2005 with a Field Scout Soil SC-900 compaction meter . Ten sites in each plot were sampled to a depth of 15 inches.

Soil moisture. Soil water storage was evaluated from volumetric soil moisture measurements taken in-row and adjacent middles to a depth of 3.5 feet at 1-foot intervals using a neutron probe. The neutron probe readings were calibrated with volumetric moisture measured from undisturbed soil cores collected at the site. Rainfall and runoff. A tipping bucket rain gauge with an 8-inch-diameter collector was used to monitor daily and cumulative rainfall at the field site. Runoff was collected at the lower end of the plots into sumps measuring 16 inches in diameter by 5 feet deep. Each sump was equipped with a device constructed from a marine bilge pump, a float switch and flow meter, to automatically record the runoff volume from the plots during storm events. During the second and third years the sampling devices were modified to collect water samples for sediment and nutrient analysis. Vine mineral nutrition. One-hundred whole leaves opposite a fruit cluster were collected from each plot at flowering in May 2003, 2004 and 2005. Petioles were separated from leaf blades, and tissue was immediately dried at 140°F for 48 hours and then sent to the ANR Analytical Laboratory for nutrient analyses. Petiole and leaf-blade tissue samples were analyzed for nitrate , ammonium , nitrogen , phosphorus , potassium , sulfur , calcium , magnesium , boron , zinc , manganese , iron and copper . Soil mineral nutrition. Composited samples from 10 soil cores taken to a depth of 1 foot were collected from the vine rows and middles at flowering as described above. Samples were air dried and sent to the ANR Analytical Laboratory for analyses. Soil samples were analyzed for pH, organic matter, cation exchange capacity , nitrate, Olsen-phosphorus, potassium, calcium, magnesium, sodium , chloride , boron and zinc. Soil microbial biomass. 

Due to the limited capacity of the laboratory, microbial biomass assays were conducted on selected treatments. Ten soil cores were collected to a depth of 1 foot and then composite samples were made from each replicate of the pre-emergence and cultivation weed-control treatments and the adjacent middles of the ‘Merced’ rye and bare treatments. Samples were collected about four times each year from November 2001 to November 2005 for a total of 14 sets of samples. Soil samples were immediately placed on ice and taken to the laboratory for soil microbial biomass carbon analysis according Vance et al. . Mycorrhizae. Roots were collected, stained and examined as previously reported on April 16, 2003, May 3, 2004, and June 2, 2005. Grape yield, fruit quality and vine growth. Fruit weight and cluster number were determined by individually harvesting 20 vines per subplot. Prior to harvest a 200-berry sample was collected from each subplot for berry weight and fruit composition. Berries were macerated in a blender and the filtered juice analyzed for soluble solids as Brix using a hand-held, temperature compensating refractometer. Juice pH was measured by pH meter and titratable acidity by titration with a 0.133 normal sodium hydroxide to an 8.20 pH endpoint. At dormancy, shoot number and pruning weights were measured from the same 20 vines. Statistical analysis. Analyses of variance were used to test the effects of cover crop, weed control and year on the vine, soil and microbial parameters, according to a split-block ANOVA model in SAS . Cover crop, weed control, year and their interactions were treated as fixed effects. The main and interactive effects of block were treated as random effects. Year was treated as May Jun Jul 26 24 22 20 18 16 B 20 18 16 14 A Bare ground Merced rye Trios 102 Fig. 3. Average soil moisture at 6 to 42 inches due to cover-crop treatments during the 2004 growing season for middles and rows ; date-by-cover-crop interaction. a repeated measure. When necessary, data were log-transformed to meet the assumption of normality for ANOVA, although untransformed or reverse transformed means are presented. Changes in soil moisture among treatments during the winter and the irrigation seasons were determined from significant treatment-date interactions. Compaction evaluation We conducted evaluations with a penetrometer each fall to determine the impact of weed-control treatments on soil compaction. Soil compaction was not significantly different at any depth in 2003 . However, in 2004 and 2005 soil compaction began to increase in the cultivation treatment compared to the other two weed-control treatments. In 2004,growing blueberries in pots soil compaction at the 4- to 7-inch depth was significantly greater in the cultivation treatment compared to the standard treatment , but not more so than in the post-emergence treatment . In 2005, the cultivation treatment had significantly greater soil compaction at the 4- to 7-inch depth than both the post emergence and standard weed-control treatments . At the 8- to 11-inch depth, soil compaction was significantly greater than the standard treatment , but not greater than in the post-emergence treatment . The blade of the cultivator passes through the soil at 2 to 6 inches deep, which may explain why greater soil compaction was measured there. Cultivations often also occurred when the soil was still moist following an irrigation, which may have contributed to the development of compacted layers over time.Moisture. 

Average, volumetric soil moisture levels at the 6- to 42-inch depth increased after the first rain events of the season, such as in winter 2002-2003 . Soil moisture declined most rapidly with ‘Merced’ rye in the middles during periods without rainfall each year , presumably due to its greater early-season growth and greater potential evapotranspiration, compared to the ‘Trios 102’ triticale. Soil moisture levels were similar between the bare and ‘Trios 102’ triticale treatments until May for all years. During the irrigation season, average soil moisture levels at the 6- to 42-inch depths were higher in rows than middles. Soil moisture in the rows and middles steadily declined during the irrigation season for all treatments during all years . Moisture levels declined most in middles with ‘Trios 102’ triticale cover during each irrigation season, presumably due to the later growth of this cover crop . In addition, the row soil-moisture levels also declined the most adjacent to ‘Trios 102’ triticale for the 2003 and 2004 irrigation seasons , but not during the 2005 irrigation season . Runoff. Total precipitation at the field trial was 7.4 inches during the 2002-2003 winter, 7.6 inches during the 2003-2004 winter and 9.9 inches during the 2004-2005 winter. A majority of the runoff was collected during December and January for the 2002-2003 and 2004-2005 winters, and February for the 2003-2004 winter. Cumulative runoff collected from individual plots during the three winters ranged from 0.02% to 3% of seasonal rainfall. Runoff was usually collected during rain events greater than 1 inch per day. Runoff was highest during the second and third years of the trial. During three consecutive winters, runoff was significantly lower in the cover crop treatments . ‘Trios 102’ triticale and ‘Merced’ rye had significantly less runoff than the bare treatment . Suspended sediment and turbidity were also significantly lower in runoff collected from the cover-crop treatments than in bare middles during winter 2004, but nutrient levels were similar among all treatments .Vines. Weed control and cover treatments did not have any significant effect on the nutritional status of the grape vines as measured by nutrient levels of the leaf petiole tissues, as determined by ANOVA. Although the nutrient levels by year were significantly different, the interactions of weed control-by-cover and weed control-bycover-by-year were not significant . Weed control and cover treatment also had no significant effect on blade nutrient content with the exception of boron and phosphate content. Vines adjacent to cover crops had significantly lower boron and phosphate levels in the leaf blade tissue than vines adjacent to bare row middles. As with the petioles, there was an absence of significance between the interaction of weed control-by-cover and weed control-by-cover-by-year for all nutrients analyzed .Soil cores indicated that most of the vine roots at this site were located under the vine row and few of the roots extended out to the row middles. This root distribution probably occurred because irrigation water was applied under the vines, and low rainfall at the site does not facilitate root growth into row middles. Thus, the lower nutrient levels in vines near cover crops may have been accentuated by irrigation effects that reduced vine root exploration of the soil to a narrow band under the vines. Since cover-crop roots probably grew into this zone there may have been competition between vines and cover crops for some nutrients. Soil. Cultivated rows had significantly lower levels of nitrate-nitrogen . Although the nutrient levels by year were significantly different, there was an absence of significance between the interaction of weed control-by-cover and weed control-by-cover-by-year . The differences observed in nitrate-nitrogen in the cultivation treatment may be due to the impact of loosening soil on water movement and leaching. Weed control treatments had occasional impacts on soil mineral nutrition in the middles, but results were inconsistent from year to year . Cover-crop treatments had no effect on soil nutrients in the rows . The most significant impacts of the vineyard floor treatments were of the cover-crop treatments on soil parameters in the middles. Soil organic matter in cover-cropped middles was higher than in bare middles each year . Cover crops affected key soil nutrients in the middles; for instance, cover crops greatly reduced nitrate-nitrogen , and to a lesser extent, extractable phosphorus , which may be beneficial in reducing loss of these nutrients in runoff during winter storms, but which also may have reduced the phosphorus content in the vines. In addition, cover crops in the middles also significantly reduced soil boron , extractable sodium and pH , and increased chloride and zinc when compared to bare soil.Soil microbial biomass. 

This conclusion is evidenced by how the explicit self-attachment to blendedness accounts for a weaker implicit Self + Harmony association, implicit harmony’s independence from any of the explicit acculturation and outcomes measures, and how bicultural individuals showed a significantly implicit stronger self-attachment to harmony relative to self + blendedness association. This line of reasoning is consistent with the argument that implicit evaluations reflect automatic affective reactions to relevant stimuli and that the harmony vs. conflict dimension is affective driven and captures the internal struggle felt within the bicultural, irrespective of the degree of overlap or similarity perceived between the two cultures. On the other hand, the blendedness vs. distance dimension of BII appears to capture elements of the explicit self-concept of the bicultural experience. This conclusion is supported by implicit blendedness’s independence from the explicit BII measure, and the correlations with explicit U.S. Identification and separation strategy,gallon nursery pot which are similar findings from Benet-Martinez and Haritatos . This line of reasoning is consistent with the fact that the blendedness vs. distance dimension is more perceptual driven and captures the performance-related elements of the acculturation experience, regardless of the degree of tension or strain felt between the two cultures and group loyalties.

The results from Study 2 do not reflect the notion that the blendedness vs. distance dimension of BII is uniquely understood in terms of the explicit self-concept, while the harmony vs. conflict dimension of BII is exclusively comprehended in light of the implicit self-concept. Both BII dimensions can exist across both levels of awareness as the current research suggests and be evaluated according to the implicit and explicit self concepts. What matters is the extent to which implicit and explicit evaluations operate independently or interactively as suggested by the dual-systems model . Research on dual-systems model posits that associative processes usually serve as a basis for explicit evaluative judgments . If the propositional implication of an implicit evaluation is in line with other relevant propositions, then it will most likely be considered as a valid basis for an evaluative judgment. However, explicit evaluative judgments exist independent of implicit evaluations, when these evaluations are discarded as a suitable source for an evaluative judgment. Thus, bicultural individuals varying in degree of BII can show assimilation or contrast effects between their implicit and explicit self-concepts based on how their implicit and explicit evaluations co-exist. This research also goes beyond the elements of biculturals who are considered high or low BII. Recall that the intersection of cultural harmony and blendedness make biculturals high BII while the merging of cultural distance and conflict make biculturalslow BII.

What about the bicultural individuals who vary independently between these two dimensions of BII. Since the two BII dimensions are orthogonal constructs, it is quite possible for a bicultural to perceive their dual cultural orientations as blended; however, feel that they are conflicted as well. For the first time the current research provides a glimpse into the possible underlying mental processes that govern how biculturals vary independently between the two BII dimensions. Results from Study 2 clearly give credence to the fact that the IAT is an optimal tool for measuring individual difference that are developed from cultural influences experienced in our daily environments. That is, the implicit evaluations the IAT measures are presumed to reflect variations in daily cultural experiences. This is corroborated by the variations in cultural experiences that biculturals encounter on a daily basis. Research has found that biculturals low in BII tend to have more negative experiences, which contribute to negative cultural associations . On the other hand, biculturals with higher levels of BII may have more positive experiences, which contribute to more positive cultural associations. Thus, any incoming cultural information that is inconsistent with the biculturals positive or negative experience may indeed produce assimilation or contrast effects between their implicit and explicit self concepts. Although Study 2 did not explicitly examine this phenomenon per se, it makes reasonable sense given that our self-concepts are “cognitive generalizations of the self, derived from past experiences, that organize and guide the processing of self-related information contained in the individual’s social experiences” .

The findings across both studies are consistent with Devos and with the image rising from modern research on acculturation and bicultural identity that individuals often find themselves immersed into multicultural surroundings and define themselves along numerous cultural boundaries and incorporate into their self concept knowledge about a variety of cultures. Going beyond the present literature, an important innovation of the present research is to provide evidence for acculturation strategies and the integration of bicultural identities into the self-concept through assessments of thoughts that cannot be consciously controlled. That is, the results of both two studies clearly demonstrate that at least under certain circumstances, cultural knowledge and/or experience can implicitly be incorporated into the self-concept. These findings are consistent with the Cultural psychology perspective which defines culture from within the individual as a socio-cognitive variable . In other words, culture and the individual are seen as interdependent rather than as independent entities . It further supports the notion that psychological acculturation can be thought of as two types of associative networks of cultural information both of which can influence an individual’s self-concept, which are shaped through repeated experiences and interactions. Finally, individuals contain, within themselves, not only various cultural meaning systems but also dual self-concepts which may be contradictory to one another. The present findings also go beyond the common assumption that psychological acculturation and bicultural identity could only be examined through conscious representations of the self-concept that are generated through an introspective reasoning process of propositions. The findings across both studies clearly showed that both psychological acculturation and bicultural identity integration can also be assessed through implicit measures such as the IAT that go beyond the restrictions of relying exclusively on explicit questionnaire methodology. Given that multicultural individuals are able to incorporate numerous cultural identities into their self-concept that go beyond explicit measures, the present research was able to contribute to the examination of the IAT-culture relationship. Unlike previous studies that failed to untangle person from culture , the present studies were able to truly conceptualize and measure culture as a socio-cognitive variable that exists within the individual rather than as independent entities. For instance, Study 1 was able to directly pair words that represented the self-concept with icons that represented various cultures together and Study 2 was able to directly pair self-concept terms with words that represented the negotiation of bicultural identity together . By using these types of IAT methodologies, the results across both studies showed support for an IAT-culture relationship. Showing support for an IATculture relationship helps corroborate the argument that cultural experience maybe what is manifested in implicit evaluations. The fact that cultural experiences helps to cultivate associative links between the self-concept and culture makes sense. Cultural experiences occur on a daily basis and help to shape the self-concept through re-occurring mundane influences that we as individuals pay no real conscious attention too.

Cultural experiences are multifaceted and are usually comprised of social categories and contexts such as national, state, city, social class, spoken language, school, occupation, friends, gender, family, ethnicity, age,greenhouse ABS snap clamp and neighborhood to name a few. The main premise of cultural experiences underlying implicit evaluations is that the experiences must occur, associations must form, and those associations must be made available . How we as social beings derive our cultural experiences has a lot to say about the ways in which we form our identities. With the influence of cross-cultural contact on the rise in the twenty-first century, new types of ethnic/cultural identities are starting to form and take shape for many individuals. Globalization based-acculturation is increasingly becoming a worldwide norm in that many individuals are now considered to be multicultural and find themselves defining their self-concepts along multiple ethnic and cultural boundaries. The argument is that although globalization based-acculturation is nothing new, it is more prevalent today than ever before and the rate at which it is expanding and influencing individuals is alarming. Today, globalization-based acculturation is driven by factors such as speed of travel and communication and have allowed for new technological marvels to penetrate every aspect of our daily lives. Computers, internet, texting, cell phones, instant messaging, and email now allow us to directly communicate and interact with individuals in other cultures that are half way around the world. As these technological devises are used everyday, they become part of our everyday cultural experiences that help shape our self-concept. Most of these new cultural encounters occur instantaneously and without conscious thought, thus allowing for many new associations to be formed and be made available in memory at an accelerated rate. Unlike twenty years ago, these new types of cultural experiences allow for individuals to expose themselves to a plethora of cultures and define their self-concepts along multiple cultural boundaries at a rate that is beyond any individual’s conscious control. Globalization-based acculturation allows for many cultural influences to act upon the individual that go beyond deliberate conscious awareness at an accelerated rate that defines the self-concept at two distinct levels of mental processing . That is why it is important to have social cognitive techniques such as the IAT capture implicit evaluations of the self-concept that are derived from cultural experiences. Limitations of study Given the important contribution of the present two studies to the literature, it is important to consider the limitations of the research. First, the cultural icons that were selected for Study 1 were not exclusively linked to each culture. Unlike the current study, Devos chose his cultural stimuli by having participants rate the extent to which each cultural icon was strongly linked to one culture and weakly linked to another culture . Each cultural icon was selected using this selection process across Devos’s two studies in order to reduce any ambiguities in linking the self-concept to the desired culture of interest. This approach was not used for the current study. By not exclusively linking the cultural icons to each culture may be a limitation in the current study because there was a potential for participants to ambiguously confuse the icons which each other. A second limitation of the current research was that the geometric icons that were used in Study 2 as a control condition were not pre-tested as being culturally neutral. Since the experimental manipulation did not work for Study 2, this was not much of a concern. Given the exploratory nature of these studies, future studies are needed to replicate these results. Future research should examine these questions in non-Latino cultural groups, who are likely to have different cultural norms, migration histories, and patterns of economic, political, and social relations in the US. Lastly, because these studies focused on the extent to which ethnic minority individuals implicitly incorporate cross-cultural knowledge and/or experiences into their self-concept, it did not include a monocultural sample. Still, future work interested in the role that cultural exposure and membership may have on the content and dynamics of the self-concept may benefit from comparisons between multicultural individuals and monoculturals . We hope that the questions and findings raised in this research resonate not only among researchers interested in implicit social cognition but also with the larger community of social, personality, and cultural researchers. As multiculturalism and globalization-based acculturation become more prevalent in the 21st century, it is important that we understand how mental processes underlie multicultural identities. Certainly, the present findings further emphasize the complexity of implicit social cognition, culture, and the self-concept, specifically showing that multicultural identities is a highly complex and layered process. In fact, as eloquently said by Devos “Research on implicit multicultural identities might provide insights into subtle, yet crucial mechanisms by which cultural knowledge is incorporated into the self-concept. It should lead to a better understanding of how cultural values, beliefs, customs, norms, and experiences shape thought, feeling, and behaviors without reflective consciousness or deliberate decisions” . Numerous studies have documented the association of early adversity with poor cardiovascular health , but knowledge of the underlying mechanisms driving this relationship is incomplete. Abnormal patterns of cardiovascular acute stress response have been operationalized as above- or below-average short-term change and slowed recovery in blood pressure or heart rate when confronting a stressor. Such responses may signal poor physiological calibration to challenges and serve as harbingers of subsequent disease development .

Concepts are also assumed to be activated by external stimuli. The presence of schemas does not mean that they influence cognition in a continuous fashion in all cases. Whether a schema is influential or not depends on the principles of availability, accessibility, salience, and applicability . Availability refers to storing associative information in memory. In order for a schema to potentially influence thought and behavior, it first has to be stored in memory . For instance, a person who plays baseball on a regular basis has stored in memory the rules of the game and the particular behaviors associated with baseball such as swing, hit the ball, and run versus finding themselves in a novel situation such as playing cricket for which they have no available information to govern their thoughts and behaviors. Accessibility refers to how easily a concept comes to mind. When a concept is more accessible it can be quickly activated and used in a particular situation. For example,drainage pot if someone is in a pool then the concept of swimming is readily accessible relative to the concept of walking. Saliency refers to the extent to which particular features of a concept stand out in the associative network relative to other features.

For instance, a woman in a room with 10 men indicates that the women’s features stand out relative to the men. Applicability refers to the particular fit between the concepts in the associative network and incoming information from the environment. For instance, the person playing baseball knows when an opposing teammate throws a ball at he/she that they are supposed to hit the ball with a bat . Individuals have different types of schemas. Of particular interests are self schemas . According to Markus self-schemas refer to “cognitive generalizations of the self, derived from past experiences, that organize and guide the processing of self-related information contained in the individual’s social experiences” In particular, the self-concept is the association between the concept self with one or more attributes . In other words, the structure of the self is a network of associations . Recent work in social cognition has identified the self-concept as a central unit in the structure of social knowledge . There are three major implications of the self-concept. First, information about the self-concept is processed faster and more efficiently, especially consistent information. Second, one retrieves and remembers information that is relevant to one’s self-concept. Third, one will tend to resist information in the environment that is inconsistent with one’s self-concept.

In essence, the social cognition framework provides an alternative approach to investigating acculturation, namely how psychological acculturation operates at an implicit level. To get a better comprehension of how psychological acculturation canoperate implicitly it is important to understand how culture is defined in relation to the self-concept. “Culture” has been traditionally operationalized as a contextual variable in cross cultural psychology. That is to say, it is often defined at the macro level as being outside of and apart from the individual . However, over the last two decades a new perspective, called “cultural psychology,” stemmed from the conceptual and methodological limitations of linking the concept of culture to the individual, a trend that dominated the traditional cross-cultural psychology perspective. Cultural psychology views culture from within rather than apart from the individual and examines how it influences actions, feelings, and thoughts. In other words, culture and the individual are seen as interdependent rather than as independent entities . Both of these approaches to culture have their strengths and weaknesses, but what is apparent about the cultural psychology studies that have been conducted over the past decade that goes beyond the cross-cultural perspective is a firm declaration that “culture is a key determinant of what is means to be a person” .

According to Hong and colleagues culture, as defined within the individual, is based upon two premises. First, culture is internalized as a loose network of specific systems of meaning which are activated by the context. Second, individuals contain, within themselves, multiple cultural meaning systems some of which may be contradictory to others. Based upon these two premises, culture can exist within the individual as a network of specific knowledge structure domains and a single individual can have various cultural meaning systems . Clearly, this type of research suggests that culture should not only be conceptualized as a situational/contextual variable which exists outside of the individual, but also at a psychological level. That is, culture can also be seen as a sociocognitive variable that exists within the individual. Capitalizing on this recent literature, psychological acculturation can be thought of as two types of associative networks of cultural information both of which can influence an individual’s self-concept, which are shaped through repeated experiences and interactions. The bidimensional model of psychological acculturation shows that culture is a multidimensional phenomenon rather than a singular construct recorded invariantly across minds. In considering culture as an integrated and highly general knowledge structure that entails relying on it continuously perhaps makes it too easy to reject the substantial influence of culture on individuals. Cultural perceptions vary across individuals and reflect the fact that individuals have unique, personal experiences of their cultural context . To better understand this phenomena perhaps it is best to use Rohner’s metaphor that compares culture to a game and people as the players . In particular, players have the choice of picking from various strategies and options, and perhaps at times even violating the rules if it serves their individual needs and purposes. That is, the degree to which individuals follow the rules varies from person to person, depending on their personal moods, preferences, and specific social context. This will often result in a great deal of within-culture heterogeneity and individual differences in the extent to which people endorse, internalize, and utilize particular rules that serve their own interests . Since culture varies across individuals within a given context, the way in which cultural information is processed from our environment is not entirely dependent on an integrated domain-general knowledge structure that entails relying on it continuously. In other words, cultural information which is cognitively processed from our environment employs bits and pieces from our ‘cultural toolbox’ to influence thoughts and behaviors.

To corroborate this fact, Hong and colleagues showed that cultural information is processed using only a small subset of an individual’s schematic knowledge structure. This subset comes to the forefront and guides the interpretation of a given environmental stimulus within bicultural individuals. The manner in which cultural information is processed is very important to consider. Recent research in social cognition has revealed a variety of dual-systems models that distinguish between how information is processed implicitly,drainage planter pot impulsively or associatively versus explicitly, reflective, or propositionally . These models suggest the extent to which implicit and explicit experiences operate independently or interactively. One important note about dual-system models is the term implicit. Implicit is often used to mean a lack of awareness or unconsciousness and includes self-regulatory processes meant to inhibit an unwarranted response . Thus, the term “implicit” refers to processes that occur outside of conscious awareness and without conscious control . According to Strack and Deitsch’s dual-systems model, thinking, and behavior are functions of two different systems of information processing, namely the reflective and the impulsive systems. In particular, the reflective system suggests that behavior is the result of propositional reasoning. For example, thinking about one’s life may lead to the conclusion “I am joyful”. Using this type of reasoning makes information accessible in the form of propositions. Propositions consist of concepts that are linked by a relation . These propositions are usually produced through introspection. The impulsive system, on the other hand, processes information by the spread of activation between concepts that are associatively linked . Associative links are activated spontaneously and are only indirectly accessible by introspection. A common assumption of research on psychological acculturation is that it involves conscious representations of the self-concept that are generated through an introspective reasoning process of propositions. Individuals are viewed as playing a consciously active role in ascribing meaning, implementing choices, pursuing goals, or initiating actions. This assumption often leads to the belief that psychological acculturation could only be examined through the process of proposition reasoning. However, research on the dual-systems model would suggest otherwise. The differences between the evaluation of implicit and explicit self-concepts should be understood in terms of their underlying mental processes . The implicit self-concept reflects automatic responses which result from the particular associations that are triggered automatically when a person encounters a relevant stimulus. The explicit self-concept, on the other hand, is best considered as evaluative judgments about the self-concept which stem from the processes of propositional reasoning.

Based on the common assumption and practice that psychological acculturation involves examining the explicit self-concept through the process of proposition reasoning, it has often been examined via questionnaires which ask respondents to describe their acculturation experiences. Although examining acculturation using a questionnaire methodology is widely accepted and a valid way to assess the topic, there are two limitations which need be addressed. First, questionnaire answers only refer to representations of the explicit self-concept that are accessible through introspection. Not all knowledge, beliefs, or attitudes of an individual are necessarily privy to introspection . Therefore, self-reports do not provide an accurate assessment of people’s thoughts and behaviors. Second, questionnaire answers are vulnerable to self-presentational biases such as social desirability, impression management, or demand characteristics. Even though self presentations biases have not been systematically evaluated in the acculturation literature, Rudmin warns us that self-report of acculturation experiences may be affected by normative demands and response bias. Due to the limitations of self-report measures and research on dual-systems model, there is a need to have access to procedures that are not restricted to these limits of explicit questionnaire measures and which are appropriate for the assessment of implicit representations of the self-concept. Over the past decade, progress has been made with the development of these implicit measures, particularly the Implicit Associate Test . The IAT’s main purpose is to measure the relative strength of automatic associations between mental representations of concepts. A basic assumption of the IAT is that if two concepts are highly associated, the sorting task will be easier when the two associated concepts share the same response key than when they share different response keys. The IAT is usually administered in a block of several trials. A trial is a one-word presentation, whereas a block is a series of trials where the category decisions are the same throughout a block. Most commonly, one IAT session consists of seven blocks, however, the number of blocks that determine one IAT session may vary depending upon the purpose of the research. An example of how the IAT works, using seven blocks, is as follows. In the first block, participants are instructed to match an item with the appropriate concept as quickly as possible. In a second block, participants are asked to distinguish between pleasant and unpleasant attributes or traits such as love and death. Thereafter, the two concepts are then paired with the pleasant or unpleasant attributes or traits . For the remaining blocks, the positions of the words are counterbalanced . The degree of association between two concepts is measured by the differences in response time to compatible and incompatible blocks. In recent years there has been much criticism in regards to what the IAT actually measures. Critics contend that the IAT measures associations that reside in the cultural environment, rather than in the person . However, proponents argue that it measures individual differences . To support this argument, Nosek and Hansen showed that the IAT has little to no relationship with cultural knowledge after accounting for common variations in explicit measures. Critics argue that a problem with the Nosek and Hansen methodology was that extrapersonal associations were used as evaluative components to measure the IAT-cultural knowledge relationship, when indeed it should be measured using a more personalized procedural approach . For example, the extrapersonal attribute-pair of pleasant/unpleasant words were associated with concept-pairs such as Black American-White American , Peanuts-Shellfish , John Kerry-George Bush , and Candy BarApple . What should have been used was a more personalized procedural approach that included personalized attribute-pairs such as I Like/I Dislike. To test this proposition, Nosek and Hansen conducted a second set of experiments to see if the IAT has any relationship with cultural knowledge after accounting for personalized changes in IAT procedures.

Another disadvantage of growing table grapes in subtropical areas is that high temperatures during ripening can inhibit anthocyanin biosynthesis in the berries from V. labrusca and hybrid cultivars . This results in inadequate fruit color, and thereby a decrease in market acceptance and the potential economic value of the commodity . The seedless table grape Selection 21, a new hybrid of V. vinifera × V. labrusca recently developed by the Grape Genetic Breeding Program of Embrapa Grape and Wine, Brazil, obtained from the cross of [Arkansas 1976 × ] × “BRS Linda,” is a clear example of a cultivar that lacks red color development when grown in subtropical regions. The plant growth regulator ethephon, an ethylene-releasing agent, has long been known to improve berry color when applied at véraison . More recently, the application of –cis-abscisic acid has also been shown to stimulate anthocyanin accumulation and thereby improve berry color . S-ABA appears to be more effective than ethephon in enhancing grape color and it has other potential benefits compared to ethephon,large pot with drainage including a shorter postharvest interval, and an exemption from tolerance in most countries.

The introduction of S-ABA as an active ingredient in a commercial plant growth regulator prompted many studies on V. vinifera cultivars under temperate climate conditions. Such studies have shown that the efficacy of S-ABA varies with the cultivar , the S-ABA concentration , the time of application and the environmental conditions . Abscisic acid is an important regulator of ripening and anthocyanin biosynthesis in grape berries . Studies have shown that exogenous application of S-ABA can significantly increase the activity of a wide range of genes involved in anthocyanin biosynthesis . Most of these studies tested the effects of a single application of S-ABA before or during véraison. However, studies of the effects of S-ABA several applications at different concentrations and timings following véraison are still needed to optimize the use of this plant growth regulator in table grape cultivation . In grapes, the anthocyanin biosynthesis pathway involves multiple steps that are controlled by MYB transcription factors, such as VvMYBA1 and VvMYBA2 . In red varieties, the VvMYBA1 gene is only expressed after véraison. Both VvMYBA1 and VvMYBA2 regulate anthocyanin biosynthesis during ripening by strictly controlling the expression of the canonical UDP-glucose:flavonoid 3-Oglucosyltransferase gene . Determining how long grape berries are competent to induce the expression of anthocyanin biosynthetic genes may help determine the optimal time, number, and frequency of S-ABA applications. Currently, little is known about the potential benefits of multiple applications, which may be desirable if a single application results in an insufficient response.

The aim of the present study was to determine the effects of S-ABA applications at different concentrations and times on the quality and biochemical properties of berries from the seedless grape Selection 21 hybrid during three growing seasons in the region of Paraná, Brazil. We evaluated a variety of parameters including: grape color development, berry phenolic profiles, and gene expression of transcriptional regulators and biosynthetic enzymes of the anthocyanin pathway after treatments with S-ABA. The results of this report indicate that two S-ABA applications during and after véraison extend the competency of grape berries to respond to ABA and induce the accumulation of anthocyanins, resulting in higher grape berry coloration. The study was conducted during three consecutive seasons in a commercial vineyard located in Marialva, state of Paraná , Brazil , using 4-year-old vines of hybrid seedless grape Selection 21 grafted onto IAC 766 Campinas root stock. According to the Köppen classification, the climate of the region is Cfa , with an average temperature below 18◦C in the coldest month and above 22◦C in the hottest month and an average annual rainfall of 1,596 mm. The region’s soil is classified as dystroferric red latosol . The vines were trained using a bilateral overhead trellis system, where vines were spaced at 2.5 m × 2.5 m , and each vine had 6.25 m2 total canopy area. Cane pruning was performed during the 2013, 2014, and 2015 seasons and was followed by application of 3% hydrogenated cyanamide to the two apical buds to induce and standardize sprouting. The number of canes per vine was evenly adjusted to 40 and the number of shoots per vine was also established to 40 .

Considering that a grape bunch of the Selection 21 weighs on average 460 g, the load per vine is 18.40 kg, which represents an estimated yield of 29.44 tons/ha. Furthermore, to avoid drifting, a non-treated vine was left as side border between two treated vines, which almost duplicated the experimental area. In each plot, all grape bunches were treated , and the bunch samples were collected from random positions at each side of the canopy to account for intracanopy variability. Control plants were not subjected to any treatment, instead, they were sprayedwith water at the same time and following the same procedures as the S-ABA treatments. The effects of applying S-ABA isomer at different concentrations and times were evaluated in terms of berry quality traits. ProTone R, the commercial growth regulator used in this study, has an active ingredient concentration of 100 g/L S-ABA. As shown in Figure 1, the initial treatments tested in the 2013 and 2014 seasons corresponded to: control or water spray, 200 mg/L S-ABA application at 7 days after véraison , 400 mg/L S-ABA application at 7 DAV, 200 mg/L S-ABA application at 7 DAV plus an additional application at 21 DAV, and 400 mg/L S-ABA at 7 DAV plus an additional application at 21 DAV. In the 2015 season, only the control and treatments of 400 mg/L S-ABA with one or two applications were performed and analyzed. Berry samples from the 2015 season were collected from each treatment at four different times: 7 DAV , 14 DAV, 28 DAV, and 35 DAV for further targeted analyses . For all seasons, a randomized complete block experimental design was used, with five treatments and three to four replicates, and with each plot consisting of one vine . Véraison was determined by measuring soluble solid content and firmness of grape berries randomly sampled in the experimental vineyard. At véraison, the mean grape SSC concentration was 9◦Bx, and 20% of the berries in more than 50% of the grape clusters presented softening . The berries presented a mean of 11◦Bx at 7 DAV, the time of the first S-ABA application, and a mean of 13◦Bx at 21 DAV, the time of the second S-ABA application. Ten grape clusters representative of each plot were marked prior to treatment application. For treatment applications, grape clusters were sprayed in the morning using a knapsack sprayer at a pressure of 568.93 psi with JA1 hollow cone nozzle tips at a volume of 800 L/ha to provide complete and uniform coverage. In addition, 0.3 mL/L of Break-Thru R a non-ionic surfactant was added to all treatments. During the trials, the standard regional cultivation practices with regard to nutrition, weed control,drainage collection pot and pest and disease management were used. Clusters of each plot were manually harvested when SSC stabilized . The clusters were cleaned, and damaged berries were discarded. Color coverage of the bunches was determined using 10 grape clusters per plot by visually rating the clusters on a scale of 1–5 using the following scale: 0–20%, 21–40%, 41–60%, 61–80%, and 81–100% coverage . The same grape clusters used for evaluating color coverage measurements were used for berry sampling. For physicochemical analyses, two berries were collected from the upper, middle, and lower portion of each grape bunch, yielding a total of 70 berries per plot. Total anthocyanins and color index of red grapes were determined in berry samples from all seasons. The following variables were analyzed only for the 2013 and 2014 seasons: color coverage, total polyphenols, and berry firmness. All physiological analyzes were performed in the Laboratory of Fruit Analysis of the Agricultural Research Center, Londrina State University, Brazil.

The total anthocyanin concentration of the berries was determined using 30 berries per plot, which were frozen and stored at −20◦C. The berry skins were removed using tweezers, taking care to remove only the skin, without pulp. The skins were washed once with water, followed by washing in deionized water and drying with absorbent paper. A 5-g skin sample was then placed in a polystyrene tube containing 50 mL of acidified methanol and stored in the dark for 48 h at room temperature. The tubes were then removed from the dark and stirred manually for 5 s. Absorbance was determined using a Genesys 10S spectrophotometer at 520 nm with the solvent as blank. In the 2015 season, three grape clusters of uniform size and at the same phenological stage were identified in each plot, and two berries were collected randomly from each bunch at each sampling time. The berry skins were removed, frozen, kept at −80◦C, and transferred to the Department of Viticulture and Enology of the University of California, Davis, CA, United States, for further analyses. The skins were then placed in liquid nitrogen and ground using a TissueLyser II . RNA was extracted using 0.5 g of ground tissue based on the protocol described by Blanco-Ulate et al. . RNA concentration and purity were determined using a NanoDrop 2000c Spectrophotometer , and RNA integrity was checked by electrophoresis on 1.5% agarose gel. Reverse transcription was performed using 1 µg of RNA and M-MLV Reverse Transcriptase according to the manufacturer’s instructions. qRT-PCRs were performed using the SYBRRGreen PCR Master Mix kit . The PCR program consisted of 70◦C for 10 min, 36 cycles at 42◦C for 2 s, and 37◦C for 50 min. VvActin was used as the reference gene and processed in parallel with the genes of interest. Gene sequences used for primer design were obtained from the GenBank of the National Center for Biotechnology Information using Primer-BLAST software . The relative levels of target gene expression were calculated using the formula 2 . The linearized values correspond to the relative gene expression within a given sample and are comparable across genes. Four biological replicates of S-ABA treated and control grape berries were used to obtain the relative gene expression data. Application of abscisic acid increased the total anthocyanin concentration in berry skins of the seedless grapeSelection 21 during the 2013 and 2014 seasons, regardless of the S-ABA concentration and time of application . However, berries that received 400 mg/L of S-ABA at 7 and 21 DAV had significantly higher, almost two to three times more, anthocyanin concentrations than other treatments. According to the CIRG, berries from control treatments had a green to a yellow color in both seasons . In 2013, berries treated with one or two applications of 200 mg/L S-ABA or one application of 400 mg/L S-ABA at 7 DAV, and those in the 2014 season that were treated with one application of 200 mg/L S-ABA developed a pink color . Remarkably, berries of the 2013 season treated with two applications of 400 mg/L S-ABA and berries of the 2014 season treated with one or two 400 mg/L S-ABA applications, developed a stronger red color . For both the 2013 and 2014 seasons, color coverage was lowest in control grapes and highest in grapes treated with two applications of 400 mg/L S-ABA. Increase in total polyphenols was evident in grapes subjected to two 400 mg/L S-ABA applications during the 2013 and 2014 seasons. These berries also presented the lowest mean berry firmness . Importantly, the increased softening due to S-ABA application did not result in higher frequency of cracked berries in any of the studied seasons. Qualitative assessment of berry cracking was performed visually. Further analyses of the effect of 400 mg/L S-ABA treatments on CIRG, total and individual anthocyanins concentrations, and gene expression of transcription factors and biosynthetic enzymes were performed with grape berries collected from the 2015 trial. Measurements of CIRG confirmed previous results obtained during the 2013 and 2014 seasons, at the time of harvest , grapes treated with two S-ABA applications had the highest CIRG values . Grape bunches from the control treatment presented pink berries , whereas those treated with one or two applications of S-ABA had red berries . As determined in the previous seasons, berries treated with 400 mg/L S-ABA also presented higher total anthocyanin content than the control at 14 and 28 DAV .

While traditional biparental populations have been effective for QTL discovery, pedigree-based analysis in multi-parental populations using FlexQTL™ has been increasingly applied, as pedigree breeding and maintenance of clones across generations are common in strawberry. Pedigree-based analysis in complex family structures has allowed the simultaneous detection of multiple QTL alleles and the quantification of their phenotypic effects across diverse genetic backgrounds, as demonstrated for the FaRPc2 locus. The use of DNA tests in breeding has been greatly enhanced by RosBREED efforts in marker development and validation. Assays for SNP detection such as kompetitive allele-specific polymerase chain reaction and high-resolution melting have become the tests of choice for breeding applications due to an abundance of SNP information from array genotyping, accuracy and ease of scoring, and resilience to crude strawberry DNA extracts. The Strawberry DNA Testing Handbook was recently developed to assist breeders in identifying published DNA tests and understanding how to apply them. This community resource is available at the Genome Database for Rosaceae and will be continually updated as existing tests are improved and new tests are published .

While locus-specific DNA tests are highly useful in parent and seedling selection for traits with simple genetics,drainage collection pot genome-wide prediction has become the strategy of choice for improving genetically complex traits in crop species. The goal is predictive, and the utility of this strategy has been demonstrated in strawberry for parent selection for yield and quality traits where it was shown that: markers are more effective than pedigrees for estimating breeding values, even when phenotypic information is present; phenotyping effort can be reduced by using trials of advanced selections as training populations; and individuals with high predicted performance can be used as parents one year early in the breeding cycle. The future of strawberry breeding research is rife with opportunities in the genomics era. In particular, candidate-gene approaches will be dramatically enhanced by the “Camarosa” reference and other octoploid genomic resources, given the ability to pinpoint sequence variations among sub-genomes and thus distinguish among homoeologous alleles. Genes involved in fruit volatile compound biosynthesis are particularly attractive targets given the importance of aroma to flavor and sweetness perception. Gene identification will, in turn, fuel the development of new DNA tests and enhance existing tests.

Questions for the future include the following: What level of genetic gains can be achieved simply by developing markers in causal genes, eliminating problems of recombination between marker and gene? Will functional characterization of genes lead to gene edited strawberries in the commercial realm? A tool that may help to uncover gene/trait associations in strawberry and help identify “missing heritability” is expression QTL . In essence, eQTL are segregating genomic regions influencing differential gene expression. With RNAseq alone, it is often difficult to discern whether changes in transcript accumulation are due to genetics, environment or stochastic effects. A recent eQTL analysis identified a subset of strawberry fruit genes whose differential expression is determined by genotype, the extent of that genetic influence, and markers that can be used for selection of desired gene-expression ranges. Thus, eQTL analyses may reveal marker/trait associations in cases where strawberry phenotypes are influenced by transcript abundance. In other cases, eQTL controlling transcripts of undetermined function can support candidate-gene discovery and trait-based gene cloning. In a recent example, simple cross-referencing of trait-QTL and eQTL markers identified a causal aroma biosynthesis gene in melon. For complex traits controlled by many loci, the area of genome-wide prediction presents a number of practically important research questions for the future. Will the newest SNP array, with its whole-genome coverage and wealth of subgenome-specific markers, help increase prediction accuracies? How large should training populations be to achieve maximum predictive power, and how many breeding cycles can be included?

Given that the vast majority of octoploid strawberry cultivars are asexually propagated, can non-additive effects be modeled to enhance predictions of clonal performance? When will low-density genotyping be affordable enough to select for complex traits in seedling populations, as opposed to selection only among parents? These questions are important and yet are very practical and applied in nature. They have been answered in other crop species, and we expect that they will soon be answered in strawberry as well.Strawberries are perennial rosette plants that form a determinate inflorescence from the apical meristem of the crown. Their axillary meristems can differentiate into either branch crowns, that are able to bear additional inflorescences, or runners. Because of these alternative fates of axillary meristems, there is a strong trade-off between flowering and runnering. Seasonal flowering strawberries produce flower initials under day lengths below a critical limit , whereas perpetual flowering strawberries produce new inflorescences continuously once induced to flower. In the diploid woodland strawberry F. vesca, the dominant progenitor of the octoploid cultivated strawberry, two classical mutants affecting flowering and runnering are known. Recessive mutations in the SF Locus and Runnering locus cause PF and runnerless phenotypes, respectively. The F. vesca homolog of TERMINAL FLOWER1 was found as a candidate gene for SFL independently by two groups, and Koskela et al. demonstrated the function of FvTFL1 as a major floral repressor that causes the seasonal flowering habit. The R locus was also recently mapped, and a mutation in a gene encoding gibberellin biosynthetic enzyme GA20-oxidase was found. This gene is highly expressed in axillary buds, and the mutated enzyme is not able to convert GA12 to GA20, a precursor of bio-active GA1. Studies in cultivated strawberry indicate at least partial conservation of the genetic pathway in woodland strawberry. Based on available data in woodland strawberry, a genetic model can be proposed . In SF genotypes, FvTFL1 integrates environmental signals to control flowering, and flower induction only occurs after the down regulation of this gene by cool temperatures below 13 °C or by short days at temperatures of 13–20 °C, whereas higher temperatures prevent flower induction by activating FvTFL.

Genes involved in the temperature regulation of FvTFL1 await elucidation, but the photoperiodic pathway is quite well understood. Under long days, the woodland strawberry homolog of CONSTANS activates FLOWERING LOCUS T1 in leaves, which leads to the upregulation of SUPPRESSOR OF THE OVEREXPRESSION OF CONSTANS1 in the shoot apex. In PF woodland strawberries that are lacking a functional FvTFL1, this FvCOFvFT1-FvSOC1 pathway promotes flowering, whereas in SF genotypes upregulation of FvTFL1 by FvSOC1 reverses the outcome of the pathway. Actual flower induction is poorly understood, but the role of FvFT3, APETALA1 , and FRUITFULL genes that are activated in the shoot apex after the down regulation of FvTFL1 by short days or cool temperature should be further explored. Another important challenge is to understand the flowering process in the context of the yearly growth cycle and to identify allelic variation that can be used for breeding new cultivars better adapted to diverse climates. Open questions include, for example, how is flower initiation and differentiation regulated? How is floral development connected to dormancy? PF cultivars are commercially quite important, but the genetic control of the trait clearly differs from PF in woodland strawberry. A major locus controlling PF was identified,10 liter pot named perpetual flowering and runnering because the PF allele also reduced runnering. Several additional studies in different crossing populations have confirmed PFRU and narrowed the chromosome region. The causal gene is not known, but several candidate genes have been suggested. Interestingly, a QTL controlling flowering time in woodland strawberry was mapped to the same region of chromosome four. These data suggest the presence of either two important flowering genes in this region or different alleles of the same gene that control PF and flowering time. Identification of the PF gene or genes in cultivated strawberry is obviously a research question of high importance both scientifically and commercially. Better understanding the trade-off between flowering and runnering is also an important area of inquiry, because it might assist plant breeders and growers in controlling the balance between vegetative and sexual reproduction. Several lines of evidence suggest that GA controls the fate of axillary meristems in strawberries. Guttridge and Thompson found that runnerless woodland strawberry mutants began to form runners after GA treatment, and similar reversion was observed in a recent mutant screen that led to the identification of suppressor of runnerless, a gene that encodes a DELLA growth repressor of the GA signaling pathway. Inhibitors of GA biosynthesis, in contrast, enhance crown branching and yield in cultivated strawberry. Furthermore, FvSOC1 was found to control runner formation and regulate the expression of several GA biosynthetic genes, including the recently identified FvGA20ox4, which likely encodes a rate limiting enzyme of the GA biosynthetic pathway in axillary buds.

These data suggest a model in which FvSOC1 activates FvGA20ox4 and possibly other GA biosynthetic genes in axillary buds, leading to high bioactive GA1 levels, degradation of SLR proteins, and runner formation .In contrast to most fruits, the fleshy tissue of Fragaria is a modified stem tip called the receptacle. The true fruits of Fragaria are dried ovaries called achenes, each of which contains a single seed. The receptacle together with attached achenes are what is typically refered to as the “fruit”. Although connected by fibrovascular strands, molecular analysis using microarrays and RNA-seq show that the achenes and receptacle exhibit asynchronous transcriptional programs that reflect differences in timing of maturation of the two tissues. Fruit set requires a sufficient percentage of fertilized achenes due to their production of auxin. Early studies showed that strawberry is non-climacteric, not appearing to respond to exogenous ethylene. However, the role of ethylene in strawberry maturation is reassessed later in this section in light of more recent data. The control of fruit set and development by plant hormones that interact and synchronize signals between the developing seed and surrounding tissues is graphically described in McAtee et al., and a comprehensive discussion of hormonal regulation of fruit ripening in non-climacteric as compared to climacteric fruit can be found in Cherian et al.. Although a complete picture of how hormonal regulation and crosstalk underlie the molecular mechanisms of development and ripening has not yet emerged in either diploid or octoploid strawberry, considerable progress has been achieved due to gene expression analyses using microarrays, RNAseq, agroinfiltration for transient gene silencing, stable transformation with reporter genes or altered expression, and virus-induced gene silencing. The high-quality diploid and octoploid strawberry genomes and associated resources now available will greatly accelerate discovery. There is still much to learn from the diploid model, and fruit development in the octoploid is likely to involve a more complex interplay of homoeologous genes. In addition, due to potential for interactions among products of homoeologs in the octoploid, careful holistic analysis of octoploid fruit development in achenes and during development remains to be accomplished by combining highly sensitive and accurate transcriptomic, proteomic, and metabolomic methods. That auxin and GAs are primary hormonal players during early development of both the receptacle and achenes is well supported by hormonal analyses of F. × ananassa and transcriptome data from F. vesca. F. vesca transcriptome data from fertilization to the large green fruit stage , were analyzed for evidence of biosynthesis and activity of most of the major hormones, as well as for IAA transport. Genes in the IAA biosynthesis pathway are actively expressed by the endosperm, and perhaps integuments, of the newly fertilized ovary, closely followed by expression of GA biosynthesis genes. This transcriptome foundation now needs to be expanded with detailed hormone metabolic and transport studies, perhaps more easily accomplished by feeding studies with the correct reference compounds and mass spectrometric analyses using larger octoploid fruit. As illustrated in Fig. 3, there are many steps in the process of fruit set and early development that still require direct study. We know that IAA levels in intact F. × ananassa fruit rise rapidly following fertilization and peak at the small green fruit stage, thereafter decreasing to low levels at the white stage and to very low, but homeostatically regulated, levels in red ripe fruit. Interestingly, separate analysis of white fruits and achenes showed that almost all the IAA measured in the intact fruit is in the achenes, with barely detectable levels in the receptacle. It has been suggested that low receptacle IAA levels are required for ABA biosynthesis to start and ripening to commence. GA1, likely the only bio-active GA in fruit, also increased after fertilization, reaching a peak in the large green stage.

Center researcher Jan Perez worked with members of the campus’s Food Systems Working Group, including UCSC Dining Services, Community Agroecology Network, and Students for Organic Solutions to develop a web-based survey designed to find out what the UCSC community thinks about food system issues. Survey results will help the groups find potential support for their work, tailor education efforts, and determine campus attitudes toward the future of sustainably produced food at UCSC. The UCSC Office of Budget and Planning implemented the survey. Asked to identify food issues and other current issues that were important to them, survey respondents ranked protecting the environment, food access for low-income people, improving food safety, improving job conditions of workers in the food system, and reducing the use of pesticides in the food system highest. The food issues that were the least important to people were limiting genetic engineering, and developing local food systems. In fact, 8% of the respondents were “unsure” about the importance of local food systems—the most people to pick that category. Food safety and nutrition were the primary interests people have in their food,container vertical farming followed by topics that encompass the impact of food production on others and the environment.

The topics garnering the least interest were the distance food travels, and the influence of large corporations. Other questions addressed interest in various “eco-labels” that represent qualities such as organic, humane treatment of animals, water quality, locally produced, and Fair Trade; whether respondents were willing to pay more for food produced with social justice criteria ; and how often people purchased Fair Trade, organic, or locally produced food. Detailed results of the survey will be presented in the next issue of The Cultivar and in a future Center Research Brief.A recent grant from PG&E’s Non-point Source Pollution Projects Fund will expand the Center’s efforts to analyze the impact of alfalfa trap crops in controlling strawberry crop damage by the western tarnished plant bug . The study also entails water quality monitoring to determine whether the use of trap crops can decrease the amount of pesticide sprays needed to control lygus populations when compared with more conventional management techniques. Center researchers Sean L. Swezey, Janet Bryer, and Diego Nieto have established the efficacy of using strips of alfalfa planted in strawberry fields to “trap” lygus, which are attracted to the alfalfa as a source of food and breeding sites . On organic farms, the strips of alfalfa are then vacuumed with a tractor-mounted “bug vac” to physically remove lygus. The new grant will expand the study of alfalfa trap crops to strawberry operations using conventional sprays to control lygus.

The study’s goal is to see whether the trap crop plantings reduce the abundance of lygus bugs and berry damage in the associated strawberry crop; reduce the number and concentration of insecticide applications made for lygus bug control; and reduce the costs of controlling lygus bugs in conventional and IPM operations.In addition, the researchers will provide growers with weekly lygus population level and crop damage estimates from the treatment plots, to use in conjunction with estimates and recommendations made by pest control advisors. This 2-year study will also include analysis of insecticide levels in runoff from strawberry farms to see whether a combination of trap cropping and potentially fewer sprays based on weekly monitoring of lygus population levels can reduce pesticide pollution originating from strawberry operations.A study on the impact of natural enemies on cabbage aphids , a significant pest of broccoli in Monterey County, appears in the February 2006 issue of Environmental Entomology . Center researcher Diego Nieto, the paper’s primary author, conducted the study on 4 organic farms in Monterey County during the summers of 2002 and 2003. He examined such factors as how the arrival date of the aphids and where on the plant they became established affected whether the broccoli head was eventually harvested . He also examined the impact of syrphid larvae , a natural enemy of aphids, on aphid abundance. Co-authors on the paper include Center director Carol Shennan, William Settle, Rachel O’Malley, Shannon Bros, and Jeffrey Honda.

Nieto’s work found aphids were most abundant on the outer leaves of most broccoli plants; these colonies did not affect infestation at harvest. This suggests that spray policies in conventionally managed systems should be adjusted to recognize aphid location as an important contributor to broccoli harvest, rather than using a zero-tolerance threshold for the presence of aphids once the broccoli head begins to form. He also found that aphid populations that became established early in the season were more likely to be suppressed by syrphid larvae and other natural enemies; hence, techniques to enhance the early establishment of natural enemy populations, such as insectary plantings, could help control aphid infestations. Social issues researcher Phil Howard’s work on Central Coast consumer preferences and concerns appears in the January–March 2006 issue of California Agriculture . Howard discusses results of focus group discussions and written surveys on consumer concerns about how their food is produced, processed, transported, and sold, and how they would prefer to get this information. The study indicates that food safety and nutrition generate the most interest, with ethical concerns and social justice issues also of interest. The majority of survey respondents indicated that food labels are their preferred source for information on these topics. Blueberries offer a potentially lucrative niche crop for organic growers that can be adapted to wholesale, retail, direct marketing, and U-pick operations. However, growing blueberries on California’s Central Coast presents challenges: most soils aren’t naturally acidic enough for blueberries to thrive, and winters don’t get cold enough to provide the chill hours that many varieties need to set a good fruit crop. To find out how blueberries can be managed in an organic Central Coast system, the Center initiated a trial of 15 high bush blueberry varieties in cooperation with Aziz Baameur, Small Farm Program Advisor for Santa Clara County’s UC Cooperative Extension office, and Mark Bolda, UCCE’s Central Coast Strawberry and Caneberry Advisor. Planted out in January 2004, the blueberries were stripped of fruit for two seasons in order to let the plants develop. This spring they’re being allowed to bear for the first time, providing the research team with an initial look at which varieties are responding best to Central Coast conditions. “It’s pretty clear that some of the more popular commercial varieties, including Jubilee, Duke, Millenia, and Ozark Blue are so far the most productive,” says Center farm manager Jim Leap, who oversees the trial. South moon is also producing well, and all of the plants are thriving even with the soil pH at 5.6—slightly more basic than is considered ideal. Leap notes that as the fruit ripens, bird predation has become an issue. He plans to install bird netting to protect the plants by early summer. The research team will also initiate a trial of a foliar iron spray this summer to see how the plants respond. “One reason you need to keep the soil’s pH low [ideally at a pH of 5] is so the plants take up iron efficiently,” explains Leap. “Testing the foliar spray will give us a good idea of the effectiveness of our current pH management strategy.”After many years of important contributions, three Center staff members are moving on to new jobs. Thomas Wittman, assistant operations manager,hydroponic vertical garden retired from the University this winter and is pursuing his own business, “Gophers Limited,” which helps gardeners and farmers control vertebrate pest damage using organic techniques. Thomas will also continue to edit a GMO news service that keeps list serve members up to date on the latest developments in the controversy over genetically modified organisms , as well as continue to serve on the Ecological Farming Association’s Board of Directors.

Marc Los Huertos, who has managed the Center’s water quality monitoring work since 1999, has accepted a tenure track faculty position at California State University Monterey Bay, where he’ll be part of the Division of Science and Environmental Policy. Marc’s teaching will focus on watershed science; he’s particularly interested in the role of nutrients in surface waters. He’ll continue to manage several water quality assessment grants for the Center over the next two years. Phil Howard, a member of the Center’s social issues research group, has accepted a position as assistant professor in the Department of Community, Agriculture, Recreation and Resource Studies at Michigan State University, where he’ll be teaching in a new graduate concentration in Community Food and Agricultural Systems. His research will involve the community in addressing questions related to the sustainability and democratization of food and agricultural systems; some topics this work will focus on include consumer interests, health impacts, and corporate consolidation. We wish Thomas, Marc and Phil all the best in their new pursuits.The Center and Global Exchange invite you to join us on a 10-day research delegation November 25 – December 4, 2006 to Havana to learn about how Cuba has become a leader in organic and urban agriculture. When the Soviet Union collapsed, the island nation of Cuba found itself cut off from the chemical fertilizers, pesticides and herbicides on which its farms had long depended. To feed its people, the Cuban government undertook an unprecedented conversion to organic and local food production. Today, much of the country’s fertilizer comes from some 175 vermicompost centers that produce more than 90,000 tons of compost annually. Thousands of urban gardens across the country help feed the island’s population, and an estimated one-third of Havana’s arable land is under cultivation. As other countries pursue conversions to more local and organic agricultural systems, many are looking toward Cuba as a model. During the 10-day delegation you will get the chance to talk with Cubans you would otherwise never meet—agronomists, government officials, community leaders, and ordinary Havana residents who are growing their own food. This is a unique opportunity to see an internationally recognized example of how to grow closer connections to our food. There was a mix of pride and apprehension as the Portuguese farmer opened his diverse enlarged home garden to a busload of foreigners. Our group was full of questions as we walked among the chickens, the regionally famous black pigs, and the early winter remains of greens, cabbage, squash, and senescing beans. Urbanites from the nearby town of Odemira recognized this small valley of Old World farmers as a place to build new relationships, or “Reciproco”—more akin to the original Japanese version of Community Supported Agriculture known as “Teikei,” where a number of farmers pool harvests to fill the weekly crate of produce for consumers. These nascent partnerships, where commitment to one another was still forming, mirrored the budding relationships of the members of the second International Symposium on Community Supported Agriculture. CSA projects are collaborations between the local community and the farmers. The community members support the farm directly for an entire season and in return receive a share in the weekly harvest. This partnership increases community involvement in food production and in the health of the local economy and environment.Our group included another Portuguese farmer who had learned about CSA for the first time when we met in Southern France almost two years ago. Now he was six months into creating a CSA project. The tour included a stop at a Lisbon storefront where Carlos’s crates of vegetables are distributed weekly. It was meaningful to see those crates—we took lots of pictures of them because they were so much like our CSA crates at home. No supermarket shelf. A simple box of mixed vegetables produced and enjoyed by people getting to know each other. The box of food, with its heads of lettuce, leeks, lemons, and rutabagas, was the common thread amongst the diverse cultures of the symposium’s attendees. What surfaced as a unique aspect of the Reciproco system, was the farmers’ reluctance to ask for a commitment from members in advance. Rather than pay for a season’s worth of food “up front,” as many CSA projects require, Reciprico members pay weekly when they pick up their crate. The farmers expressed trepidation over not being able to provide for their members, and the members might have been afraid of not getting what they paid for. However, for the attendees at the symposium, supporting the transition to a more conscious relationship between urban and rural was the bottom line.