Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of developing wild type pea seeds and those expressing anti-ABA antibodies

ABSTRACT: ABA regulates in plants a wide range of developmental events, mediates responses to environmental stress and is necessary to proceed through seed maturation and to acquire desiccation tolerance and dormancy. Immuno-modulation is a suitable means to study ABA functions during seed maturation. Anti-ABA single chain antibody was expressed in pea seed driving LeB4-promoter (Saalbach et al., High-level expression of a single chain Fv fragment (scFv) antibody in transgenic pea seeds J. Plant Physiol. 2001 158: 529-533), which produced only a weak phenotype with slightly decreased seed weight, globulin/albumin and total nitrogen content (aABA line 16 cultivar Erbi). In another approach with a stronger, improved USP-promoter used to express the anti-ABA antibody in pea seeds a different phenotype emerged (aABA line 7, cultivar Eifel). In this line individual seed weight increased by 20 to 30% together with higher globulin and albumin content. To dissect the aABA phenotype at the molecular level, a search for genes with differential expression patterns in transgenic plant versus wild type seeds has been performed using 6k-oligo microarray analysis. cDNA probes were prepared from RNA isolated from embryo of developing seeds of wild type (12, 18, and 22 DAP) and transgenic aABA plants (12, 18, and 22 DAP), which correspond to the transition phase of seed development, and 6k-oligo microarray.

ORGANISM(S): Pisum sativum

SUBMITTER: Ruslana Radchuk

PROVIDER: E-MEXP-1555 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Seed development is a complex process controlled by many factors, including genetics, plant growth regulators, and the environment. Understanding the different compositions and abundances of proteins in various types of seeds at different developmental stages is fundamental for improving seed quality and enhancing nutritional and food security. In this study, we applied label-free quantitative proteomics to analyze round and wrinkled pea seeds at five different growth stages: 4, 7, 12, 15 days after anthesis (DAA), and at maturity. Wrinkled peas had lower starch content (29.5%) compared to round peas (~47-55%). Proteomic analysis identified 3,659 protein groups, of which approximately 21-24% proteins were shared across growth stages. Relatively more proteins were identified during early seed development compared to later stages, and the protein profiles were also drastically different between early and late stages. Statistical analysis identified 735 significantly different proteins between wrinkled and round seeds, regardless of the growth stage. These detected proteins were characterized into different functional classes including metabolic enzymes, proteins involved in protein biosynthesis and homeostasis, carbohydrate metabolism, vesicle trafficking, cell division, and cell wall organization. Cell division-related proteins were more abundant in the early stages of seed development, whereas storage proteins and proteins implicated in lipid metabolism were more abundant at the later stages. Wrinkled seeds had a significantly lower abundance of starch-branching enzyme than did round seeds. This enzyme is involved in the biosynthesis of the amylopectin component of starch. Seed storage proteins such as legumin and a form of albumin (PA2) accumulated more in round pea genotypes, whereas vicilin was more abundant in the wrinkled pea genotype. In summary, this study can enhance our understanding of pea seed development, highlighting key differences in protein profiles between round and wrinkled pea seeds.

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Transcription profiling of developing wild type pea seeds and those expressing anti-ABA antibodies

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