陆地棉果枝夹角关键基因GhPDF1的功能鉴定及育种演化分析

doi:10.3864/j.issn.0578-1752.2026.09.002

摘要/Abstract

摘要：

【目的】果枝夹角是决定棉花株型的关键性状。GhPDF1在果枝夹角极端材料中的表达呈现显著差异，开展GhPDF1的生物学功能鉴定，系统解析其优异等位变异，为棉花株型改良及机采棉品种选育提供重要基因资源和理论依据。【方法】通过同源比对，获取PDF1在陆地棉等25个物种中的序列信息，并进行系统进化分析；采用双酶切法构建GhPDF1的病毒诱导的基因沉默（VIGS）载体，并转化陆地棉，明确GhPDF1在调控果枝夹角中的作用；利用418份陆地棉种质资源的重测序数据，挖掘GhPDF1内部的单核苷酸多态性（SNP）位点，并用Sanger测序验证关键SNP位点；分析GhPDF1内部优异等位变异在我国不同育种时期品种中的演变情况。【结果】构建陆地棉PDF1与不同物种同源基因的系统发育树，发现它与锦葵科亲缘关系最近，距离禾本科较远；通过分析该基因在果枝夹角极端材料的表达量，发现其在大夹角材料中低表达，在小夹角材料中高表达；组织表达分析表明，GhPDF1在雌蕊、花瓣和茎等组织中呈现高丰度表达。VIGS沉默GhPDF1结果显示，相较空载体对照植株，沉默植株果枝夹角连接处的细胞数量显著减少约170个、果枝夹角显著增大8.2°，表明该基因是调控陆地棉果枝夹角的关键基因；通过对等位变异GhPDF1^GG和GhPDF1^CC的果枝夹角表型分析，发现GhPDF1^GG的果枝夹角极显著小于GhPDF1^CC，且随着育种时期的不断推进，该优异等位变异的频率从92%上升到98%。【结论】在陆地棉中沉默GhPDF1，可以促进果枝连接处细胞数量减少，导致果枝夹角显著增大，其基因内部优异等位变异GhPDF1^GG的频率随着育种年份的变化呈现逐步上升演变趋势。

关键词: 陆地棉, 果枝夹角, GhPDF1, 病毒诱导的基因沉默, 等位变异

Abstract:

【Objective】The fruit branch angle is a key determinant of cotton plant architecture. Previous studies revealed differential expression of GhPDF1 in materials with extreme fruit branch angles. This study aimed to functionally characterize GhPDF1 and systematically dissect its superior allelic variations, thereby providing genetic resources and a theoretical basis for improving plant architecture and breeding machine-harvestable cotton varieties.【Method】The sequences of PDF1 homologs from 25 species, including upland cotton (Gossypium hirsutum), were obtained through homology alignment for phylogenetic analysis. A virus-induced gene silencing (VIGS) vector targeting GhPDF1 was constructed using double digestion and transformed into upland cotton to investigate its role in regulating the fruit branch angle. Single nucleotide polymorphisms (SNPs) within GhPDF1 were identified using resequencing data from 418 upland cotton accessions, with key SNPs validated by Sanger sequencing. The distribution of superior allelic variations of GhPDF1 was analyzed across Chinese cotton varieties from different breeding eras.【Result】A phylogenetic tree of the PDF1 gene from Gossypium hirsutum and its homologous genes from different species revealed that it is most closely related to Malvaceae, while being distantly related to Poaceae. Expression analysis showed that GhPDF1 was lowly expressed in large‑angle materials but highly expressed in small‑angle materials. Tissue‑specific expression profiling revealed high transcript levels in pistils, petals, and stems. VIGS‑mediated silencing of GhPDF1 significantly reduced the cell number at the fruit branch junction by approximately 170 and increased the fruit branch angle by 8.2° compared with empty‑vector controls, demonstrating its crucial role in regulating fruit branch angle. Phenotypic comparison between two allelic variants, GhPDF1^GG and GhPDF1^CC, indicated a significantly smaller branch angle in GhPDF1GG carriers. Moreover, the frequency of this superior allele (GhPDF1^GG) increased from 92% to 98% over successive breeding periods.【Conclusion】Silencing GhPDF1 in upland cotton reduces cell proliferation at the branch junction, leading to a significant increase in the fruit branch angle. The superior allelic variant GhPDF1^GG has been progressively enriched during modern cotton breeding in China.

Key words: Upland cotton, fruit branch angle, GhPDF1, virus-induced gene silencing, allelic variation

张文姣, 魏佳智, 周亚荣, 杨浩然, 郭荣欣, 马俊峰, 杨嘉辉, 王彩香, 宿俊吉. 陆地棉果枝夹角关键基因GhPDF1的功能鉴定及育种演化分析[J]. 中国农业科学, 2026, 59(9): 1836-1847.

ZHANG WenJiao, WEI JiaZhi, ZHOU YaRong, YANG HaoRan, GUO RongXin, MA JunFeng, YANG JiaHui, WANG CaiXiang, SU JunJi. Functional Identification and Breeding Evolution Analysis of the Key Gene GhPDF1 for Fruit Branch Angle in Upland Cotton[J]. Scientia Agricultura Sinica, 2026, 59(9): 1836-1847.

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引物名称 Primer name	引物序列 Primer sequence (5′-3′)	产物长度 Product length (bp)	用途 Purpose
GhPDF1-F	GTGGCAGTCCACCAACTACT	400	克隆 Cloning
GhPDF1-R	GCAAGCTTGAAGAGACGAGC	400	克隆 Cloning
MQGhPDF1-F	gactagtGTGGCAGTCCACCAACTACT	415	沉默 VIGS
MQGhPDF1-R	aggcgcgccGCAAGCTTGAAGAGACGAGC	415	沉默 VIGS
qGhPDF1-F	TGTGGGTTTTGGTTGCTGC	122	实时荧光定量PCR qRT-PCR
qGhPDF1-R	TGAAGGGGGAGTTCCAGCAT	122	实时荧光定量PCR qRT-PCR
SNPGhPDF1-F	CCGCCCATGTTCCATGAGT	319	SNP验证 SNP validation
SNPGhPDF1-R	GCAGCAACCAAAACCCACAT	319	SNP验证 SNP validation