Bulked Segregant RNA-Seq Analysis of Pollinated Pistils Reveals Genes Influencing Spikelet Fertility in Rice

Prezygotic isolation is important for successful fertilization in rice, significantly affecting yield. This study focused on F_(5:6) generation plants derived from inter-subspecific crosses(Nipponbare × KDML105) with low(LS) and high seed-setting rates(HS), in which normal pollen fertility was observed. However, LS plants showed a reduced number of pollen grains adhering to the stigma and fewer pollen tubes reaching the ovules at 4-5 h post-pollination, compared with HS plants. Bulked segregant RNA-Seq analysis of pollinated pistils from the HS and LS groups revealed 249 and 473 differentially expressed genes(DEGs), respectively. Kyoto Encyclopedia of Genes and Genomes analysis of the HS and LS-specific DEGs indicated enrichment in metabolic pathways, pentose and glucuronate interconversions, and flavonoid biosynthesis. Several of these DEGs exhibited co-expression with pollen development genes and formed extensive clusters of co-expression networks. Compared with LS pistils, enzyme genes controlling pectin degradation, such as OsPME35 and OsPLL9, showed similar expression patterns, with higher levels in HS pistils pre-pollination. Os02g0467600, similar to cinnamate 4-hydroxylase gene(CYP73), involved in flavonoid biosynthesis, displayed higher expression in HS pistils post-pollination. Our findings suggest that OsPME35, OsPLL9, and Os02g0467600 contribute to prezygotic isolation by potentially modifying the stigma cell wall(OsPME35 and OsPLL9) and controlling later processes such as pollen-stigma adhesion(Os02g0467600) genes. Furthermore, several DEGs specific to HS and LS were co-localized with QTLs and functional genes associated with spikelet fertility. These findings provide valuable insights for further research on rice spikelet fertility, ultimately contributing to the development of high-yielding rice varieties.

Development of new aromatic rice lines with high eating and cooking qualities

Rice is the staple food for about half of the world’s population.Preferred by consumers,aromatic rice is a special type of rice with great commercial value.Cooking and eating qualities and aroma are the major grain qualities favored by most consumers.Currently,most of the available aromatic varieties have low yields and some undesirable agronomic traits.Thus,there is an urgent need to develop better aromatic rice varieties.This work aims to identify rice germplasm lines that have good grain quality and to develop new varieties with desirable traits.Thirty-six out of 188 germplasm lines were found to have betaine aldehyde dehydrogenase 2 (badh2) controlling the aroma and were analyzed for their 2-acetyl-1-pyrroline (2AP) contents.Then,17 of those lines were found to have alleles for low amylose content and low gelatinization temperature,controlled by waxy and starch synthase Ⅱa (SSⅡa),respectively,suggesting that they are aromatic rice lines with high cooking and eating qualities.A total of 158 F_(7)recombinant inbred lines (RILs) generated from five crosses of the selected germplasm lines were planted for phenotypic and yield observations,resulting in 27 F_(8)RILs selected for yield evaluation and genotyping.Finally,four out of the seven F_(9)aromatic RILs showed high yield,high 2AP production,and low amylose content,in agreement with their genotypes.The other three F9RILs were aromatic rice lines with high amylose content and high yield.Because consumer preferences for grain quality vary depending on regions and ethnic groups,the high-yielding aromatic RILs generated from this study can be used to increase the yield of Thai rice and to raise market value and farm profits.