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Deciphering the dynamic expression network of fiber elongation and the functional role of the GhTUB5 gene for fiber length in cotton based on an introgression population of upland cotton
Journal of Advanced Research ( IF 11.4 ) Pub Date : 2024-08-04 , DOI: 10.1016/j.jare.2024.08.004
Jianjiang Ma 1 , Liupeng Yang 2 , Yuanyue Dang 3 , Kashif Shahzad 2 , Jikun Song 2 , Bing Jia 2 , Li Wang 1 , Juanjuan Feng 2 , Nuohan Wang 4 , Wenfeng Pei 1 , Man Wu 1 , Xuexian Zhang 2 , Jinfa Zhang 5 , Jianyong Wu 1 , Jiwen Yu 6
Affiliation  

Introduction

Interspecific introgression between Gossypium hirsutum and G. barbadense allows breeding cotton with outstanding fiber length (FL). However, the dynamic gene regulatory network of FL-related genes has not been characterized, and the functional mechanism through which the hub gene GhTUB5 mediates fiber elongation has yet to be determined.

Methods

Coexpression analyses of 277 developing fiber transcriptomes integrated with QTL mapping using 250 introgression lines of different FL phenotypes were conducted to identify genes related to fiber elongation. The function of GhTUB5 was determined by ectopic expression of two TUB5 alleles in Arabidopsis and knockout of GhTUB5 in upland cotton. Yeast two-hybrid, split-luciferase and pull-down assays were conducted to screen for interacting proteins, and upstream genes were identified by yeast one-hybrid, dual-LUC and electrophoretic mobility shift assays.

Results

The 32,612, 30,837 and 30,277 genes expressed at 5, 10 and 15 days postanthesis (dpa) were grouped into 19 distinct coexpression modules, and 988 genes in the MEblack module were enriched in the cell wall process and exhibited significant associations with FL. A total of 20 FL-QTLs were identified, each explaining 3.34–16.04 % of the phenotypic variance in the FL. Furthermore, several FL-QTLs contained 15 genes that were differentially expressed in the MEblack module including the tubulin beta gene (TUB5). Compared with the wild type, the overexpression of GhTUB5 and GbTUB5 in Arabidopsis suppressed root cell length but promoted cellulose synthesis. Knockout of GhTUB5 resulted in longer fiber lines. Protein-based experiments revealed that GhTUB5 interacts with GhZFP6. Additionally, GhTUB5 was directly activated by GhHD-ZIP7, a homeobox-leucine zipper transcription factor, and its paralogous gene was previously reported to mediate fiber elongation.

Conclusion

This study opens a new avenue to dissect functional mechanism of cotton fiber elongation. Our findings provide some molecular details on how GhTUB5 mediates the FL phenotype in cotton.


中文翻译:


基于陆地棉渗入群体破译棉花纤维伸长的动态表达网络和 GhTUB5 基因对棉花纤维长度的功能作用


 介绍


Gossypium hirsutumG. barbadense 之间的种间渗入允许培育具有出色纤维长度 (FL) 的棉花。然而,FL 相关基因的动态基因调控网络尚未表征,枢纽基因 GhTUB5 介导纤维伸长的功能机制尚未确定。

 方法


使用 250 个不同 FL 表型的渗入系对 277 个发育中的纤维转录组进行共表达分析,并与 QTL 定位相结合,以鉴定与纤维伸长相关的基因。通过在拟南芥中异位表达两个 TUB5 等位基因和敲除陆地棉中敲除 GhTUB5 来确定 GhTUB5 的功能。进行酵母双杂交、分裂荧光素酶和沉降测定以筛选相互作用的蛋白质,并通过酵母单杂交、双 LUC 和电泳迁移率变化测定鉴定上游基因。

 结果


将开花后 5 、 10 和 15 天 (dpa) 表达的 32,612 、 30,837 和 30,277 个基因分为 19 个不同的共表达模块,MEblack 模块中的 988 个基因在细胞壁过程中富集,并与 FL 表现出显著关联。共鉴定了 20 个 FL-QTL,每个 QTL 解释了 FL 中 3.34-16.04% 的表型变异。此外,几个 FL-QTL 包含 15 个在 MEblack 模块中差异表达的基因,包括微管蛋白 β 基因 (TUB5)。与野生型相比,拟南芥GhTUB5GbTUB5 的过表达抑制了根细胞长度,但促进了纤维素合成。敲除 GhTUB5 导致纤维线更长。基于蛋白质的实验表明,GhTUB5 与 GhZFP6 相互作用。此外,GhTUB5 被同源盒-亮氨酸拉链转录因子 GhHD-ZIP7 直接激活,其旁系同源基因先前报道介导纤维伸长。

 结论


本研究为剖析棉花纤维伸长的功能机制开辟了一条新途径。我们的研究结果提供了有关 GhTUB5 如何介导棉花 FL 表型的一些分子细节。
更新日期:2024-08-04
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