Rice Research Institute, Yunnan Agricultural University, Kunming 650201, China.
College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, China.
Int J Mol Sci. 2023 Jun 26;24(13):10640. doi: 10.3390/ijms241310640.
Pollen development and its fertility are obligatory conditions for the reproductive success of flowing plants. () is known to be preferentially expressed and may play critical role in developing pollen. A 31-bp InDel was identified as a unique variation and was shown to be responsible for the expression of downstream gene in our previous study. In this study, to analyze the changes of gene expression triggered by 31-bp InDel during pollen development, two vectors () were constructed and then stably introduced into rice. Histochemical and quantitative real-time PCR (qRT-PCR) analysis of transgenic plants showed that 31-bp deletion drastically reduced the expressions of downstream genes, including both and in rice panicle at booting stage, especially that of . The transcriptome profile of two types of panicles at booting stage revealed a total of 1028 differentially expressed genes (DEGs) between 31-bp In and 31-bp Del transgenic plants. Further analyses showed that 397 of these genes were significantly enriched for the 'metabolic process' and 'binding'. Among them, nineteen genes had a strong relationship with starch and sucrose metabolism and were identified as candidate genes potentially associated with the starch accumulation in rice pollen, which that was also verified via qRT-PCR. In summary, 31-bp InDel plays a crucial role not only in the regulation of downstream genes but in the expression of sucrose-starch metabolizing genes in multiple biological pathways, and provides a different regulation mechanism for sucrose metabolism in pollen.
花粉的发育和育性是流动植物生殖成功的必要条件。已知()优先表达,可能在花粉发育中发挥关键作用。我们之前的研究中发现了一个 31bp 的 InDel,它是一个独特的变异,被证明负责下游基因的表达。在这项研究中,为了分析 31bp InDel 在花粉发育过程中触发的基因表达变化,构建了两个载体(),然后稳定地导入水稻。对转基因植物的组织化学和定量实时 PCR(qRT-PCR)分析表明,31bp 缺失极大地降低了下游基因的表达,包括水稻穗期启动阶段的()和(),特别是()的表达。两种穗期的转录组图谱分析显示,31-bp In 和 31-bp Del 转基因植物之间共有 1028 个差异表达基因(DEGs)。进一步分析表明,其中 397 个基因在“代谢过程”和“结合”中显著富集。其中,有 19 个基因与淀粉和蔗糖代谢有很强的关系,被鉴定为与水稻花粉中淀粉积累相关的候选基因,这也通过 qRT-PCR 得到了验证。总之,31-bp InDel 不仅在下游基因的调控中起着关键作用,而且在多个生物途径中蔗糖-淀粉代谢基因的表达中也起着关键作用,为花粉中的蔗糖代谢提供了一种不同的调控机制。