Project description:Pea (Pisum. sativum L.) is a traditional and important edible legume that can be sorted into grain pea and vegetable pea according to their harvested maturely or not. Vegetable pea by eating the fresh seed is becoming more and more popular in recent years. These two type peas display huge variations of the taste and nutrition, but how seed development and nutrition accumulation of grain pea and vegetable pea and their differences at the molecular level remains poorly understood. To understand the genes and gene networks regulate seed development in grain pea and vegetable pea, high throughput RNA-Seq and bioinformatics analysis were used to compare the transcriptomes of vegetable pea and grain pea developing seed. RNA-Seq generated 18.7 G raw data, which was then de novo assembled into 77,273 unigenes with a mean length of 930 bp. Functional annotation of the unigenes was carried out using the nr, Swiss-Prot, COG, GO and KEGG databases. There were 459 and 801 genes showing differentially expressed between vegetable pea and grain pea at early and late seed maturation phases, respectively. Sugar and starch metabolism related genes were dramatically activated during pea seed development. The up-regulated of starch biosynthesis genes could explain the increment of starch content in grain pea then vegetable pea; while up-regulation of sugar metabolism related genes in vegetable pea then grain pea should participate in sugar accumulation and associated with the increase in sweetness of vegetable pea then grain pea. Furthermore, transcription factors were implicated in the seed development regulation in grain pea and vegetable pea. Thus, our results constitute a foundation in support of future efforts for understanding the underlying mechanism that control pea seed development and also serve as a valuable resource for improved pea breeding.
Project description:12plex_pea_2013_02 - 12plex_pea_2013_02_f - What is the effect of a moderate water stress on seed filling (reserve accumulation) and nitrogen remobilisation in pea (Pisum sativum) - Pea plants (genotype Cameor) were subjected to a moderate water stress at the beggining of the seed filling period (12 Days After Pollination) of the second flowering node for a period of 8 days. Samples were collected from Well Watered (WW) plants at the beginning of the stress imposition (point A, T=0), and from Water-Stressed (WS) and WW control plants at the end of the drought period (point B, T=+8). Samples named SEED consisted of seeds from the pod of the second flowering node (seed-WW-A, seed-WW-B and Seed-WS-B). Samples named LEAF consisted of the leaves and stem sections from the two vegetative nodes below the first flowering node (leaf-WW-A, Leaf-WW-B and Leaf-WS-B). Each sample consited of a pool of 3 individual plants and 4 repetitions per condition were carried out.
Project description:12plex_pea_2013_02 - 12plex_pea_2013_02_g - What is the effect of a moderate water stress on seed filling (reserve accumulation) and nitrogen remobilisation in pea (Pisum sativum) - Pea plants (genotype Cameor) were subjected to a moderate water stress at the beggining of the seed filling period (12 Days After Pollination) of the second flowering node for a period of 8 days. Samples were collected from Well Watered (WW) plants at the beginning of the stress imposition (point A, T=0), and from Water-Stressed (WS) and WW control plants at the end of the drought period (point B, T=+8). Samples named SEED consisted of seeds from the pod of the second flowering node (seed-WW-A, seed-WW-B and Seed-WS-B). Samples named LEAF consisted of the leaves and stem sections from the two vegetative nodes below the first flowering node (leaf-WW-A, Leaf-WW-B and Leaf-WS-B). Each sample consited of a pool of 3 individual plants and 4 repetitions per condition were carried out.
Project description:During seed growth, sugar and nitrogen compounds confer regulatory control on storage activities. Thus, seed storage production could be regulated by the supply of nutrients. In order to improve nitrogen flux into the embryo, transgenic pea lines were created where ADP-glucose pyrophosphorylase (AGP) from Pisum sativum has been repressed by RNAi approach in the seeds under control of the seed-specific LeB4 promotor (Bäumlein et al. Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within legumin box is essential for tissue-specific expression of a legumin gene. Plant J 1992 2: 233-239). The plastidial enzyme AGP catalyzes the reaction of glucose-1-phosphate and ATP to pyrophosphate and ADP-glucose, which is the substrate for starch synthase. The AGP activity and transcript levels were strongly decreased in three independent transgenic lines. Repression of AGP results in a wrinkled seed phenotype obviously due to transient accumulation of free sugars during maturation. Mature seeds have reduced starch content whereas the protein concentration is higher due to increased fractions of albumins and globulins. Repression of AGP interferes with storage protein metabolism and alters fluxes of nitrogen during seed growth. The influence of decreased AGP on altered gene expression in developing cotyledons was analysed using a 6k-Oligo-microarray. Ps6kOLI1 microarray hybridization were performed using three independent biological replicates of four developmental stages (20, 25, 30 and 35 DAP) from seeds of the transgenic line iAGP-3.
Project description:In order to change assimilate partitioning during seed maturation a bacterial phosphoenolpyruvate carboxylase (PEPC) has been expressed in V. narbonensis under control of the LegB4 promoter. Transgenic bean seeds accumulate app. 20% more protein and have higher seed weight. Physiological and biochemical analyses indicate a shift of metabolic fluxes from sugars/starch to organic acids/free amino acids suggesting increased anaplerotic carbon flow. This indicates that the availability of carbon acceptors limits seed protein synthesis. Gene expression was analysed in growing embryos using macro-arrays containing 5,548 seed-specific pea genes. Radioactive labeled cDNA probes were prepared from RNA isolated from embryo of developing seeds of wild type and transgenic PEPC plants of V. narbonensis (15, 17, 19, 21, 25, and 30 DAP) and hybridized to cDNA macroarrays.