OsPRR95 corresponds to Arabidopsis PRR, AtPRR5 or AtPRR9.A report has revealed that triple mutant prr 9–11 prr 7–10 prr 5–10 Arabidopsis exhibit better salt, drought and cold tolerance than wild type, and thus suggested that PRR5, PRR7 and PRR9 are involved in the diurnal cold stress-initiating stress response by mediating the cyclic expression of stress response genes, including DREB1/CBF.Additionally, Mesembryanthemum crystallinumCSP1, which is a class of pseudo-response regulator-like proteins, co-localizes with calcium-dependent protein kinasein the nucleus of NaCl-stressed ice plants, suggesting that it may be regulated by McCDPK1 through reversible phosphorylation.According to the MSU7 database, LOC_Os05g38710, the novel CaM1 target, is annotated as lipin, and the mRNA sequence of LOC_Os05g38710 is annotated as phosphatidate phosphatase.A report has demonstrated that the N- and C-terminal regions of mammalian lipin protein share sequence similarity to yeast PAH1.Phosphatidate phosphatase is the enzyme that converts phosphatidic acid to diacylglycerol and Pi.In Phaseolus vulgaris cotyledons, phosphatidate phosphatase is stimulated by Ca2+ or CaM with Ca2+, and a possible role of Ca2+-second-messenger in membrane-lipid degradation initiation has been suggested.Therefore,hydroponic nft system its identification as a CaM-interacting protein herein suggests that Ca2+/CaM stimulates phosphatidate phosphatase via direct binding.
By protein functional association analysis of each of these CIPs, the GO terms enriched in each set of resulting associated proteins that matched those from OsCam1–1 affected salt-responsive DEGs are presented in Fig.10.Matched GO terms revealed interacting protein candidates that potentially regulate various cellular processes represented by each enriched GO term of the OsCam1–1 affected salt-responsive DEGs.Cadmium , one of the most toxic heavy metals for both plants and humans, accumulates in the human body through the food chain and causes serious health problems.In recent years, the accumulation of Cd in rice grains has become an important agricultural problem in Japan because the Cd content of rice grains sometimes exceeds the limit proposed by the Codex Alimentarius Commission.In addition, Cd intake from rice accounts for about one-half of the intake from food in Japan according to the National Institute of Health Sciences.Therefore, new technologies for reducing the Cd content in rice grains are urgently required.Although the mechanism underlying the uptake and translocation of Cd in plants is not completely understood, some irontransporters, such as OsIRT1 and OsIRT2, are reported to uptake Cd as well as Fe.AtNramp3 and AtNramp4 from Arabidopsis, which belong to the Nramp metal transporter family, function as Fe and Cd transporters.Rice has seven Nramp genes , and OsNramp1 has been reported to function as an Fe transporter.In this study, we investigated the possibility that OsNramp1 also transports Cd.
Full-length OsNramp1 was amplified by RT-PCR using total RNA prepared from hydroponically grown rice shoots.The subcellular localization of OsNramp1 was determined by monitoring the expression of an OsNramp1::GFP fusion protein in onion epidermal cells transformed by DNA particle bombardment.GFP, contained in the vector pH7FWG2, was fused to the 3’-terminus of OsNramp1 using the Gateway system.To test the growth of OsNramp1-expressing yeast, full-length OsNramp1 cDNA was inserted into the expression vector pYH23.The construct was then introduced into yeast strain ycf1 using the lithium acetate method.Ycf1 lacks the YCF1 transporter, which functions in the compartmentation of Cd into vacuoles.Yeast cells transformed with empty pYH23 were used as a control.The transformed yeast cells were grown in synthetic defined medium and spotted onto SD agar containing CdCl2.To measure the metal content of the OsNramp1-overexpressing rice, plants were grown in Cd-contaminated soil for 6 months in a greenhouse.Harvested leaf blades were dried at 70˚C for 1 week.Sample digestion and measurement of the metal content were performed as described previously , except that the digestion time and temperature were changed to 2 h at 230˚C.Manned space exploration missions deploy technologies and products that mitigate crew-safety concerns and that assist with mission accomplishment.These technologies are continuously evaluated for relevance and cost, a term that accounts for launch mass, drawn power, volumetric size, useful product life, astronaut utility, etc.This evaluation is important, because space missions are inherently expensive; every unit mass of payload that is launched into space necessitates the launch of an additional 99 units of mass.Hence, there is an interest in novel technologies that simultaneously decrease cost, reduce risk and increase the probability of mission success.Typically, the cost of these new technologies is reduced through in situ resource utilization which consists of harnessing materials located at a mission’s destination.This paper investigates how current biological techniques and future synthetic biology progress can meet several of the above-mentioned needs.The work reviews existing biological processes to demonstrate that they already constitute a competitive yet non-traditional technology that is capable of processing volatiles and waste resources readily available on two representative space missions in a way that reduces the launch mass of propellant, food and raw material for three-dimensional printing, and also overcomes the decreased product shelf-life of a common therapeutic.
The paper employs these reviewed processes in designs for natural and artificially enhanced biological manufacturing strategies that can be leveraged to saThisfy space input availability and output-desirability constraints.The work then analyses methodological feasibility, technique versatility and the costs and yields of feed stocks and constituents, and compares possible future ‘space synthetic biology’ advances to other new aerospace technologies.Although a novel technique, synthetic biology has already been tapped for its potential to eliminate plastic waste, enrich food, monitor pollution and chemicals and be an ISRU tool.This paper furthers these forays and widens the scope of the technology by indicating its capacity for extensive product applicability in space despite the severe input limitations imposed by the space environment.Of these candidate missions, the fourth warrants an analysis of applicable bio-production techniques for two reasons: a lengthy total time spent on board a spacecraft that is comparable to the long residence time of the asteroid-investigation mission, and an extremely lengthy stay on Mars that makes it vital to explore all technologies that could reduce risk, decrease launch mass and manufacture products with a short shelf-life.Given its possible precursor status for the Martian mission, the lunar mission will also be examined for bio-manufacturing benefits.Representative values of the masses of crew-produced wastes, which serve as potential resources for biology based designs, are listed in table 1 for the previously stated Martian- and lunar-manned space exploration missions.Further inputs for these two missions can be drawn from the Mars atmosphere or the permanently shadowed craters on the south pole of the Moon, respectively,and the Martian soil or lunar regolith, respectively.
The tables suggest that carbon dioxide and nitrogen are somewhat plentiful resources for biology applications over the course of a Mars voyage and stay.However, these resources are significantly scarcer on a Moon mission.Yet, if large enough excavators and bioreactors are deployed , there should be enough of these resources extracted to test the viability of biological techniques prior to a Mars voyage.Hydrogen and oxygen may also be available for bio-manufacturing on both missions as a result of the electrolysis of polar water, but it is expected that water availability will be reduced given its priority to support crew life.If required, hydrogen can be transported to Mars and also stored until use, but this process is considered somewhat difficult and problematic.On the Moon however, hydrogen is already present.Oxygen may also be harvested from the Martian soil or the lunar regolith with post-excavation processing.Hence, bio-production applications for Mars and the Moon need to take as inputs: carbon dioxide, nitrogen , hydrogen and oxygen,nft channel ordered here by their availability.This resource set of elements and simple compounds can conceivably support biological systems because its constituent elements form a subset of the main elements required for life, namely carbon, hydrogen, nitrogen, oxygen, phosphorus and sulfur.The latter two elements are not readily available on both Martian and lunar missions, although sulfur is present on the Moon alone.Soil-based metal resources from Mars and regolith-derived metal resources from the Moon are not considered suitable for biology application in this paper.A cost-based ordering of the items required for a Mars or Moon mission cannot be compiled, because cargo manifests for these future missions are still in flux.Moreover, at this early stage of mission planning, the form of the cost metric itself and the relative weightings it contains is somewhat unclear.This is attributable to changing technology readiness-levels and disparate evaluation criteria.Nevertheless, four targets quantifiably stand out for biological production.Fuel, for instance, is currently projected to be ‘about two-thirds of the mass on an Earth-to-Mars-to Earth mission cost-effective [extraterrestrial]-produced propellant could decrease the mass that must be lifted from Earth by a factor of two to three’.Food is another necessary target, as evidenced by crew meals constituting the bulk of a recent supply mission to the International Space Station.Bio-polymers are a third target, because plastics are included in the list of feed stock materials that can be used for three dimensional printing.The three-dimensional printing of structures to manufacture a spacecraft in space can decrease roughly 30% of the craft’s launch mass by reducing the supporting structural material that is required, and additive manufacturing can also reduce the launch mass cost associated with storing a multitude of spacecraft spare parts.Because the 30% number presumes a launch of necessary printer media, the extraterrestrial production of raw material for three dimensional printing, e.g.bio-polymers, can achieve even greater mass reductions.Further savings can also be realized by deploying additive manufacturing for other purposes, such as the construction of habitats, rocket engine parts, sample containers, spacecraft electronic platforms, etc.Lastly, the accelerated expiration of pharmaceuticals induced by space radiation necessitates the on-demand synthetic manufacture of such pharmaceuticals on long-duration missions.In this paper, a versatile drug to treat infection and pain symptoms, e.g.aspirin, acetaminophen, etc., is targeted for biosynthesis.It is envisioned that this drug will be manufactured when desired by astronauts using bacteria that are activated from a frozen state.The bacteria will not themselves ‘expire’ from space radiation because of storage in a small, lead-lined container while inactive; bacterial spores and rock colonizing eukaryotes can survive with little protection in space for between 1.5 and 6 years.
Quality control of astronaut-activated bacteria can be performed through portable gene sequencers that are in development, and that are already being contemplated for use in space exploration.The choice of four targets outlined in this section is further justified by their inclusion in the list of needs presented in §1 for which NASA seeks promising new technologies.Sections 3–6 confirm the feasibility and benefits of producing each of these desirable endpoints with contemporary biological techniques.Hence, the design problem that is tackled in this paper: design biology processes to go from the inputs listed in the left column of figure 2 to the outputs listed in the right column of figure 2, using the fewest number of intermediates,organisms and steps, with the greatest possible commonality of such intermediates, organisms and steps, and with the goal of substantially reducing launch mass and increasing product shelf-life.The availability of only a few input elements and simple compounds coupled with the predefined desirability of various output products constrain space biology designs.The current state of the technology requires design options to include, as a first step, those organisms that already use the same resources on the Earth.Thereafter, the outputs of these organisms can serve as inputs to other organisms.The yields of such modular designs can be analysed and then improved upon with bioengineering and genetic modification techniques.As synthetic biology matures over the coming decades, it may be possible to build designer organisms from scratch that directly manufacture the desired products efficiently.Because carbon dioxide and nitrogen compounds are the dominant available resources, organisms that harness these resources and the yields and efficiencies at which they do so are of prime importance.Further, the outputs of these organisms will be useful as either the desirable targets of figure 2 or as feed stock intermediates to obtain these targets.Thus, we summarize in electronic supplementary material, table S1 the mechanisms of action and the outputs produced by organisms that take in carbon dioxide, as detailed by.Electronic supplementary material, table S2 provides a similar summary of organisms that use and produce various nitrogen compounds; these organisms also play a role in the microbial nitrogen cycle.A greedy design approach involves employing the lowest-energy carbon dioxide fixation process from electronic supplementary material, table S1, which exists in methanogens and acetogens.Conveniently, the responsible pathway in these organisms, the Wood –Ljungdahl pathway, requires the input of hydrogen and the presence of anoxic conditions.