Transcriptome analysis identifies differentially expressed genes in the progenies of a cross between two low phytic acid soybean mutants

Phytic acid (PA) is a major antinutrient that cannot be digested by monogastric animals, but it can decrease the bioavailability of micronutrients (e.g., Zn and Fe). Lowering the PA content of crop seeds will lead to enhanced nutritional traits. Low-PA mutant crop lines carrying more than one mutated gene (lpa) have lower PA contents than mutants with a single lpa mutant gene. However, little is known about the link between PA pathway intermediates and downstream regulatory activities following the mutation of these genes in soybean. Consequently, we performed a comparative transcriptome analysis using an advanced generation recombinant inbred line with low PA levels [2mlpa (mips1/ipk1)] and a sibling line with homozygous non-mutant alleles and normal PA contents [2MWT (MIPS1/IPK1)]. An RNA sequencing analysis of five seed developmental stages revealed 7945 differentially expressed genes (DEGs) between the 2mlpa and 2MWT seeds. Moreover, 3316 DEGs were associated with 128 metabolic and signal transduction pathways and 4980 DEGs were annotated with 345 Gene Ontology terms related to biological processes. Genes associated with PA metabolism, photosynthesis, starch and sucrose metabolism, and defense mechanisms were among the DEGs in 2mlpa. Of these genes, 36 contributed to PA metabolism, including 22 genes possibly mediating the low-PA phenotype of 2mlpa. The expression of most of the genes associated with photosynthesis (81 of 117) was down-regulated in 2mlpa at the late seed developmental stage. In contrast, the expression of three genes involved in sucrose metabolism was up-regulated at the late seed developmental stage, which might explain the high sucrose content of 2mlpa soybeans. Furthermore, 604 genes related to defense mechanisms were differentially expressed between 2mlpa and 2MWT. In this study, we detected a low PA content as well as changes to multiple metabolites in the 2mlpa mutant. These results may help elucidate the regulation of metabolic events in 2mlpa. Many genes involved in PA metabolism may contribute to the substantial decrease in the PA content and the moderate accumulation of InsP3–InsP5 in the 2mlpa mutant. The other regulated genes related to photosynthesis, starch and sucrose metabolism, and defense mechanisms may provide additional insights into the nutritional and agronomic performance of 2mlpa seeds.

(209) Development of Germinability and Desiccation Tolerance in Lettuce Seeds

Germinability and desiccation tolerance are important attributes that seeds acquire during their development. The timing in the expression of these characteristics is important to understand how environmental conditions affecting the mother plant influence seed quality. Lettuce plants (cv. Tango) were cultivated in the greenhouse. Seed germination, under light and darkness, was evaluated in fresh and dry seeds at 3, 5, 7, 9, 11, 13, 15, and 17 days after flowering (DAF). Desiccation was performed ≈1 h after harvest by placing the seeds at 25 °C and ≈53% RH. The seed moisture level after desiccation decreased from ∼14% for 3 DAF seed to ∼7% for 7 DAF seed, and then remained constant until the last sampling. Seeds achieved maximum dry weight (physiological maturity) at ∼13 DAF. Germination of fresh seeds increased from 0% at 3DAF to ∼80% at 5 DAF, reaching 100% at 7 DAF. Dry seeds did not germinate when they were 3 or 5 DAF. Seeds at 7 DAF had ∼10% germination and at 9 DAF ∼100%. When germinated in the dark, an increase in germination from 0% in fresh seeds at 3 DAF to 50% germination at 5 DAF was observed. However, seeds at 9 DAF had dark germination values that decreased to 0% and increased again to ∼70% germination at 13 DAF. Dry seeds had no dark germination until 7 DAF, with variable and low germination (below 20%) until 11 DAF, then germination reached a maximum of ∼55% at 13 DAF and decreased to below 10% at 17 DAF. According to these results, lettuce seed germinability and desiccation tolerance were reached sooner than physiological maturity. In the dark, germination of fresh seeds presented a curve with two peaks suggesting that, depending on the seed developmental stage, two different physiological mechanisms restrict dark germination.