作物学报2025，Vol.51Issue(8)：2060-2076,17.DOI:10.3724/SP.J.1006.2025.44197
陆地棉杜松烯合酶基因GhCDN10的特征及其在棉酚合成中功能分析
Characterization of GhCDN10 encoding cadinene synthase and its involvement in gossypol biosynthesis pathway in Gossypium hirsutum
摘要
Abstract
Gossypol is a major terpenoid compound in cotton(Gossypium hirsutum),predominantly stored in pigment glands distributed throughout the plant.Cadinene synthase is a key enzyme involved in the gossypol biosynthetic pathway.Due to its toxicity,large quantities of cottonseed products cannot be fully utilized for human consumption.Therefore,the development of new cultivars featuring"plants with high gossypol content but seeds with none or low levels of gossypol"is essential to maximize the value of the cotton industry.Based on RNA-Seq data from glanded and glandless cotton,we cloned GhCDN10,a member of the cadinene synthase gene family,and conducted DNA sequence analysis,including linkage disequilibrium(LD),SNP effect prediction,haplotype construction,and association analysis.We also performed expression profiling,virus-induced gene silencing(VIGS),and subcellular localization assays.GhCDN10 comprises seven exons,and its flanking intergenic regions(IRs)contain 533 cis-acting elements from 63 classes.Re-sequencing data revealed a SNP frequency of 7.22 SNPs kb-1 in GhCDN10,with anucleotide diversity(π)value of 0.22595 and an LD decay distance of approximately 100 kb.Nineteen SNPs were identified within the gene body,forming nine haplotypes.Although these SNPs were not directly associated with gossypol content,cluster analysis based on them could partially distinguish low-gossypol from normal accessions.The GhCDN10 protein contains two conserved domains(PF01397 and PF03936)and conserved motifs such as DDTYD and DDVAE,characteristic of cadinene syn-thase.Subcellular localization of the GFP fusion protein showed that GhCDN10 is localized to both the plasma membrane and nucleus.Comparative analysis revealed slight variations in exon-intron structure and motif arrangement among GhCDN10 orthologs from 17 species.RNA-Seq data showed significantly higher GhCDN10 expression in glanded cotton compared to glandless cotton.This gene is highly expressed in roots of glanded cotton but shows low or no expression in flowers and fibers.Following VIGS treatment,GhCDN10 expression decreased to 21.3%of the control level.Microscopic analysis and HPLC quan-tification revealed significantly reduced gland numbers and gossypol content in VIGS plants,highlighting the critical role of GhCDN10 in gossypol biosynthesis and pigment gland development.This study provides new insights into the gossypol biosyn-thetic pathway and identifies GhCDN10 as a promising candidate gene for the development of glandless cotton germplasm through genetic engineering.关键词
陆地棉(Gossypium hirsutum)/杜松烯合酶/GhCDN10/棉酚/基因克隆/VIGSKey words
Gossypium hirsutum/Delta-cadinene synthase/GhCDN10/gossypol/cloning/VIGS引用本文复制引用
薛晓菲,戴云静,李熙林,丁艳艳,王翔,雷长英,韩焕勇,贺道华..陆地棉杜松烯合酶基因GhCDN10的特征及其在棉酚合成中功能分析[J].作物学报,2025,51(8):2060-2076,17.基金项目
本研究由国家自然科学基金项目(32301956),西北农林科技大学科研启动费项目(2452023053)和财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-15-25)资助.This study was supported by the National Natural Science Foundation of China(32301956),the PhD Start-up Research Fund of Northwest A&F University(2452023053),and the China Agriculture Research System of MOF and MARA(CARS-15-25). （32301956）
