Ma Qi-Feng, Wu Chun-Hui, Wu Man, Pei Wen-Feng, Li Xing-Li, Wang Wen-Kui, Zhang Jinfa, Yu Ji-Wen, Yu Shu-Xun
State Key Laboratory of Cotton Biology, Institute of Cotton Research of CAAS, Anyang, Henan 455000, China.
Department of Plant and Environmental Sciences, New Mexico State University, Las Cruces, NM 88003, USA.
Sci Rep. 2016 Apr 14;6:24485. doi: 10.1038/srep24485.
To investigate the molecular mechanisms of fiber initiation in cotton (Gossypium spp.), an integrated approach combining transcriptome, iTRAQ-based proteome and genetic mapping was taken to compare the ovules of the Xuzhou 142 wild type (WT) with its fuzzless-lintless (fl) mutant at -3 and 0 day post-anthesis. A total of 1,953 mRNAs, 187 proteins, and 131 phosphoproteins were differentially expressed (DE) between WT and fl, and the levels of transcripts and their encoded proteins and phosphoproteins were highly congruent. A functional analysis suggested that the abundance of proteins were mainly involved in amino sugar, nucleotide sugar and fatty acid metabolism, one carbon pool for folate metabolism and flavonoid biosynthesis. qRT-PCR, Western blotting, and enzymatic assays were performed to confirm the regulation of these transcripts and proteins. A molecular mapping located the lintless gene li3 in the fl mutant on chromosome 26 for the first time. A further in-silico physical mapping of DE genes with sequence variations between fl and WT identified one and four candidate genes in the li3 and n2 regions, respectively. Taken together, the transcript abundance, phosphorylation status of proteins at the fiber initiation stage and candidate genes have provided insights into regulatory processes underlying cotton fiber initiation.
为了研究棉花(棉属物种)纤维起始的分子机制,采用了一种整合的方法,将转录组、基于iTRAQ的蛋白质组和遗传图谱相结合,以比较徐州142野生型(WT)及其无绒无絮(fl)突变体在开花后-3天和0天的胚珠。WT和fl之间共有1953个mRNA、187个蛋白质和131个磷酸化蛋白质差异表达(DE),转录本及其编码的蛋白质和磷酸化蛋白质的水平高度一致。功能分析表明,蛋白质丰度主要参与氨基糖、核苷酸糖和脂肪酸代谢、叶酸代谢的一碳池以及类黄酮生物合成。进行了qRT-PCR、蛋白质免疫印迹和酶活性测定,以证实这些转录本和蛋白质的调控。分子图谱首次将fl突变体中的无绒基因li3定位在第26号染色体上。对fl和WT之间具有序列变异的DE基因进行进一步的电子物理图谱分析,分别在li3和n2区域鉴定出1个和4个候选基因。综上所述,纤维起始阶段的转录本丰度、蛋白质磷酸化状态和候选基因为棉花纤维起始的调控过程提供了见解。