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Yuhai Cui - One of the best experts on this subject based on the ideXlab platform.
Mutagenesis of seed Storage Protein genes in Soybean using CRISPR/Cas9BMC research notes, 2019Co-Authors: Vi Nguyen, Jun Liu, Chen Chen, Yuhai CuiAbstract:
Soybean seeds are an important source of vegetable Proteins for both food and industry worldwide. Conglycinins (7S) and glycinins (11S), which are two major families of Storage Proteins encoded by a small family of genes, account for about 70% of total soy seed Protein. Mutant alleles of these genes are often necessary in certain breeding programs, as the relative abundance of these Protein subunits affect amino acid composition and soy food properties. In this study, we set out to test the efficiency of the CRISPR/Cas9 system in editing soybean Storage Protein genes using Agrobacterium rhizogenes-mediated hairy root transformation system. We designed and tested sgRNAs to target nine different major Storage Protein genes and detected DNA mutations in three Storage Protein genes in soybean hairy roots, at a ratio ranging from 3.8 to 43.7%. Our work provides a useful resource for future soybean breeders to engineer/develop varieties with mutations in seed Storage Proteins.
genetic mapping and validation of the loci controlling 7s α and 11s a type Storage Protein subunits in soybean glycine max l merrTheoretical and Applied Genetics, 2018Co-Authors: Jeffrey D Boehm, Vi Nguyen, Rebecca M Tashiro, Dale Anderson, Chun Shi, Lorna Woodrow, Yuhai CuiAbstract:
Four soybean Storage Protein subunit QTLs were mapped using bulked segregant analysis and an F2 population, which were validated with an F5 RIL population. The Storage Protein globulins β-conglycinin (7S subunit) and glycinin (11S subunits) can affect the quantity and quality of Proteins found in soybean seeds and account for more than 70% of the total soybean Protein. Manipulating the Storage Protein subunits to enhance soymeal nutrition and for desirable tofu manufacturing characteristics are two end-use quality goals in soybean breeding programs. To aid in developing soybean cultivars with desired seed composition, an F2 mapping population (n = 448) and an F5 RIL population (n = 180) were developed by crossing high Protein cultivar ‘Harovinton’ with the breeding line SQ97-0263_3-1a, which lacks the 7S α′, 11S A1, 11S A2, 11S A3 and 11S A4 subunits. The Storage Protein composition of each individual in the F2 and F5 populations were profiled using SDS-PAGE. Based on the presence/absence of the subunits, genomic DNA bulks were formed among the F2 plants to identify genomic regions controlling the 7S α′ and 11S Protein subunits. By utilizing polymorphic SNPs between the bulks characterized with Illumina SoySNP50K iSelect BeadChips at targeted genomic regions, KASP assays were designed and used to map QTLs causing the loss of the subunits. Soybean Storage Protein QTLs were identified on Chromosome 3 (11S A1), Chromosome 10 (7S α′ and 11S A4), and Chromosome 13 (11S A3), which were also validated in the F5 RIL population. The results of this research could allow for the deployment of marker-assisted selection for desired Storage Protein subunits by screening breeding populations using the SNPs linked with the subunits of interest.
Ken Pendarvis - One of the best experts on this subject based on the ideXlab platform.
Proteome rebalancing in transgenic Camelina occurs within the enlarged proteome induced by β-carotene accumulation and Storage Protein suppressionTransgenic Research, 2017Co-Authors: Monica A Schmidt, Ken PendarvisAbstract:
Oilseed crops are global commodities for their oil and Protein seed content. We have engineered the oilseed Camelina sativa to exhibit increased Protein content with a slight decrease in oil content. The introduction of a phytoene synthase gene with an RNAi cassette directed to suppress the Storage Protein 2S albumin resulted in seeds with an 11–24 % elevation in overall Protein. The phytoene synthase cassette alone produced enhanced β-carotene content of an average 275 ± 6.10 μg/g dry seed and an overall altered seed composition of 11 % less Protein and comparable nontransgenic amounts of both oil and carbohydrates. Stacking an RNAi to suppress the major 2S Storage Protein resulted in seeds that contain elevated Protein and slight decrease in oil and carbohydrate amounts showing that Camelina rebalances its proteome within an enlarged Protein content genotype. In both β-carotene enhanced seeds with/without RNAi2S suppression, the seed size was noticeably enlarged compared to nontransgenic counterpart seeds. Metabolic analysis of maturing seeds indicate that the enhanced β-carotene trait had the larger effect than the RNAi2S suppression on the seed metabolome. The use of a GRAS (generally regarded as safe) β-carotene as a visual marker in a floral dip transformation system, such as Camelina , might eliminate the need for costly regulatory and controversial antibiotic resistance markers. β-carotene enhanced RNAi2S suppressed Camelina seeds could be further developed as a rapid heterologous Protein production platform in a nonfood crop leveraging its enlarged Protein content and visual marker.
Richard Thompson - One of the best experts on this subject based on the ideXlab platform.
Transcriptional regulation of Storage Protein synthesis during dicotyledon seed fillingPlant and Cell Physiology, 2008Co-Authors: Jérôme Verdier, Richard ThompsonAbstract:
Seeds represent a major source of nutrients for human and animal livestock diets. The nutritive value of seeds is largely due to Storage products which accumulate during a key phase of seed development, seed filling. In recent years, our understanding of the mechanisms regulating seed filling has advanced significantly due to the diversity of experimental approaches used. This review summarizes recent findings related to transcription factors that regulate seed Storage Protein accumulation. A framework for the regulation of Storage Protein synthesis is established which incorporates the events before, during and after seed Storage Protein synthesis. The transcriptional control of Storage Protein synthesis is accompanied by physiological and environmental controls, notably through the action of plant hormones and other intermediary metabolites. Finally, recent post-genomics analyses on different model plants have established the existence of a conserved seed filling process involving the master regulators (LEC1, LEC2, ABI3 and FUS3) but also revealed certain differences in fine regulation between plant families.
In situ expression of two Storage Protein genes in relation to histo-differentiation at mid-embryogenesis in Medicago truncatula and Pisum sativum seedsJournal of Experimental Botany, 2005Co-Authors: Mona Abirached-darmency, M.r. Abdel-gawwad, Geneviève Conejero, Jean-luc Verdeil, Richard ThompsonAbstract:
The seed consists of several layers of specialized cell-types that divide and differentiate following a highly regulated programme in time and space. A cytological approach was undertaken in order to study the histo-differentiation at mid-embryogenesis in Medicago truncatula as a model legume, and in Pisum sativum using serial sections of embedded immature seed. Little published information is available about seed development in Medicago species. The observations from this study revealed a number of distinctive features of Medicago seed development and differentiation. Transfer cells, involved in nutrient transfer to the embryo, were clearly identified in the thin-walled parenchyma of the innermost integument. Histological Schiff–naphthol enabled carbohydrate accumulation to be followed in the different seed compartments, and revealed the Storage Protein bodies. Non-radioactive mRNA in situ hybridization, was carried out using mRNA probes from two highly expressed genes encoding the major vicilin and legumin A Storage Protein types. The timing of mRNA expression was related to that of the corresponding Proteins already identified.
Derek J Bewley - One of the best experts on this subject based on the ideXlab platform.
the role of abscisic acid in germination Storage Protein synthesis and desiccation tolerance in alfalfa medicago sativa l seeds as shown by inhibition of its synthesis by fluridone during developmentJournal of Experimental Botany, 1995Co-Authors: Nanfei Xu, Derek J BewleyAbstract:
Abscisic acid and osmoticum maintain maturation and Protein synthesis of developing alfalfa embryos, individually and in combination. The in situ environment of developing alfalfa zygotic embryos is rich in ABA and low in osmotic potential. When ABA synthesis was inhibited by treating the pods with fluridone at an early stage of development, the seeds which subsequently developed contained low amounts of ABA, but had a similar osmotic potential as untreated control seeds. The reduced ABA in seeds from fluridone-treated pods did not change the morphology except the colour of seeds, nor did it induce viviparous germination or affect Storage Protein synthesis. However, two non-Storage Proteins which were synthesized in control seeds during early to mid-development were absent from fluridone-treated seeds. Control seeds containing these two Proteins were desiccation-tolerant, whereas the fluridone-treated seeds which lacked them were desiccation-intolerant, at least until the deposition of Storage Proteins was nearly complete. Culture of isolated embryos on nutrient medium induced germination and curtailed Storage Protein synthesis in the embryos. Addition of either ABA or osmoticum to the nutrient medium prevented germination and maintained Storage Protein synthesis. When fluridone was added along with osmoticum, germination occurred, but Storage Protein synthesis was maintained
contrasting Storage Protein synthesis and messenger rna accumulation during development of zygotic and somatic embryos of alfalfa medicago sativa lPlant Physiology, 1992Co-Authors: Joan E Krochko, Saroj K Pramanik, Derek J BewleyAbstract:
During development on hormone-free media, somatic embryos pass through distinct morphological stages that superficially resemble those of zygotic embryo development (globular, heart, torpedo, cotyledonary stages). Despite these similarities, they differ from zygotic embryos in the extent of cotyledonary development and the patterns of synthesis and quantitative expression of seed-specific Storage Proteins (7S, 11S, and 2S Proteins). Alfin (7S) is the first Storage Protein synthesized in developing zygotic embryos (stage IV). The 11S (medicagin) and 2S (Low Molecular Weight, LMW) Storage Proteins are not detectable until the following stage of development (stage V), although all three are present before the completion of embryo enlargement. Likewise, the 7S Storage Protein is the first to be synthesized in developing somatic embryos (day 5). Medicagin is evident by day 7 and the LMW Protein by day 10. In contrast to zygotic embryos, alfin remains the predominant Storage Protein in somatic embryos throughout development. Not only are the relative amounts of medicagin and the LMW Protein reduced in somatic embryos but the LMW Protein is accumulated much later than the other Proteins. Quantification of the Storage Protein mRNAs (7S, 11S, and 2S) by northern blot analysis confirms that there are substantial differences in the patterns of message accumulation in zygotic and somatic embryos of alfalfa (Medicago sativa). In zygotic embryos, the 7S, 11S, and 2S Storage Protein mRNAs are abundant during maturation and, in particular, during the stages of maximum Protein synthesis (alfin, stages VI and VII; medicagin, stage VII; LMW, stage VII). In somatic embryos, the predominance of the 7S Storage Protein is correlated with increased accumulation of its mRNA, whereas the limited synthesis of the 11S Storage Protein is associated with much lower steady-state levels of its message. The mRNA for the LMW Protein is present already by 3 days after transfer to hormone-free media, yet that Protein is not evident on stained gels until day 10. Thus, both transcriptional and posttranscriptional events appear to be important in determining the Protein complement of these seed tissues. On the basis of Storage Protein and mRNA accumulation, mature (14 days) somatic embryos most closely resemble stage VI zygotic embryos. The results of the developmental comparison also suggest that the patterns of synthesis of the individual Storage Proteins (7S, 11S, or 2S) are regulated independently of each other during embryogenesis in alfalfa.
Iain E. P. Taylor - One of the best experts on this subject based on the ideXlab platform.
Storage Protein gene expression in zygotic and somatic embryos of interior sprucePhysiologia Plantarum, 1993Co-Authors: Barry S. Flinn, Dane R. Roberts, Craig H. Newton, David R. Cyr, Fiona B. Webster, Iain E. P. TaylorAbstract:
Storage Protein gene expression was compared between zygotic and somatic embryos of interior spruce (Picea glauca/engelmanii complex). Somatic embryos, grown on medium containing 40 μM or 10 μM abscisic acid (ABA), and zygotic embryos accumulated IIS legumin, 7S vicilin and 2S albumin Storage Proteins. Zygotic embryos displayed a rapid, transient period of Storage Protein accumulation, while somatic embryos differentiated on 40 μM ABA displayed a more prolonged, gradual accumulation, with some accumulation still evident after 9 weeks of maturation. Somatic embryos on 10 μM ABA accumulated Storage Proteins initially, but these were rapidly degraded as the embryos germinated precociously. Legumin, albumin and vicilin transcripts were detectable in torpedo stage zygotic and somatic embryos, and increased during embryo development. Transcript levels in zygotic embryos increased during cotyledon development, but following maximum dry weight accumulation and moisture loss, transcripts declined rapidly to low levels. In contrast, somatic embryos on 40 μM ABA had high transcript levels for a prolonged period. These levels were still present after 9 weeks of maturation. A decline in Storage Protein transcripts similar to zygotic embryos was apparent following a mild drying treatment. These results suggest that a decline in Storage Protein transcripts is stimulated by embryo drying during the later stages of conifer embryogenesis. Low levels of Storage Protein transcripts also appeared in somatic embryos on 10 μM ABA, but declined during precocious germination. Osmotic stress induced Storage Protein and Storage Protein transcript accumulation. This could be partially inhibited by inclusion of the ABA biosynthetic inhibitor, fluridone. However, endogenous ABA levels did not differ significantly between embryos cultured in the presence or absence of fluridone.