南方农业学报2023，Vol.54Issue(12) ：3475-3487.DOI:10.3969/j.issn.2095-1191.2023.12.003

宁春4号×河东乌麦F6:7家系籽粒蛋白质性状及候选基因分析

Grain protein traits and candidate genes analysis in F6:7 pedigrees derived from Ningchun No.4×Hedong black wheat

陈佳静 毛馨缘 王新杰 李清峰 刘彩霞 张雪婷 赵补全 亢玲 李前荣 王掌军
南方农业学报2023，Vol.54Issue(12) ：3475-3487.DOI:10.3969/j.issn.2095-1191.2023.12.003
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  • NSTL
  • 万方数据

宁春4号×河东乌麦F6:7家系籽粒蛋白质性状及候选基因分析

Grain protein traits and candidate genes analysis in F6:7 pedigrees derived from Ningchun No.4×Hedong black wheat

陈佳静 1毛馨缘 1王新杰 1李清峰 1刘彩霞 1张雪婷 1赵补全 1亢玲 2李前荣 2王掌军1
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作者信息

  • 1. 宁夏大学农学院,宁夏银川 750021
  • 2. 宁夏农林科学院农作物研究所,宁夏银川 750001
  • 折叠

摘要

[目的]对宁春4号和河东乌麦杂交、自交F6:7家系籽粒蛋白质性状及候选基因进行分析,以期发掘与籽粒蛋白质性状相关的候选基因,为小麦籽粒蛋白质性状遗传改良提供理论参考.[方法]以蛋白质性状差异较大的2个亲本(宁春4号与河东乌麦)及其杂交、自交的551个F6:7家系为材料,对小麦籽粒蛋白质性状(粗蛋白含量、湿面筋含量和沉降值)进行方差分析、相关分析及聚类分析,并对高、低值株系进行转录组测序、差异表达基因(DEGs)功能注释、信号通路富集分析及与籽粒蛋白质性状相关基因鉴定.[结果]籽粒蛋白质相关性状在F6:7家系中均出现明显分离,分布均呈连续正态分布.粗蛋白含量、湿面筋含量和沉降值的平均值分别为15.64％、33.31％和39.06 mL,介于双亲该性状值之间,其中,超中亲比例为29.04％～77.31％,超高亲比例为7.08％～39.56％;变异系数排序为沉降值(10.99％)>湿面筋含量(6.07％)>粗蛋白含量(5.70％),且三者之间呈极显著(P<0.001)正相关.551个F6:7家系被聚为四大类群(Ⅰ～Ⅳ),分别包括148、125、171和107个家系,其中,类群Ⅳ的平均粗蛋白含量(16.87％)、湿面筋含量(35.96％)和沉降值(45.02 mL)均最高,为优异类群.根据高、低值株系的转录组测序数据,从中筛选出4368个DEGs,其中,970个为上调表达,3398个为下调表达.GO功能注释结果显示,小麦籽粒蛋白质性状差异表达主要发生在细胞区域,涉及细胞膜结构和细胞器等组分,同时也发生在细胞连接等部位,通过调节结合、催化等分子功能,引起细胞过程、代谢过程、防御反应和应激反应为主的系列生物过程的协同变化.KEGG信号通路富集结果显示,DEGs显著(P<0.05)富集到9条代谢通路,其中筛选出4条与小麦籽粒蛋白质性状密切相关的代谢通路及其DEGs,10个DEGs与组氨酸代谢、21个与淀粉和蔗糖代谢、8个与酪氨酸代谢、8个丙氨酸和天冬氨酸及谷氨酸代谢相关,其中有7个上调表达.实时荧光定量PCR验证结果与RNA-Seq结果基本一致.[结论]小麦籽粒蛋白质性状属多基因控制的数量性状遗传,F6:7家系出现许多超亲类型,其中沉降值具有较大改良潜力.类群Ⅳ为籽粒蛋白质性状优异类群,可用于蛋白质性状优异品种的育种实践.基于富集功能分析筛选出组氨酸代谢、淀粉和蔗糖代谢、酪氨酸代谢、丙氨酸和天冬氨酸及谷氨酸代谢4条通路与小麦籽粒蛋白质性状调控机制有关,并鉴定出7个与籽粒蛋白质性状相关的候选基因.

Abstract

[Objective]Analysis of grain protein traits and candidate genes in F6:7 pedigrees of Ningchun No.4 and He-dong black wheat through the cross and self-cross was carried out to discover candidate genes associated with grain pro-tein traits,which will provide theoretical reference for genetic improvement of grain protein traits in wheat.[Method]551 F6:7 pedigrees deriving from the cross and self-cross between two parents(Ningchun No.4 and Hedong black wheat)which showed significant differences in grain protein traits,were used as materials.Variance analysis,correlation analysis and cluster analysis were performed to analyze the grain protein traits(crude protein content,wet gluten content and sedi-mentation value)and transcriptome sequencing on high and low value strains,functional annotation,signal pathway enrichment analysis,and identification of genes related to grain protein traits were carried out on differentially expressed genes(DEGs).[Result]The grain protein-related traits in the F6:7 pedigrees were obviously separated,and the distribution was continuous normal distribution.The average values of crude protein content,wet gluten content and sedimentation value were 15.64％,33.31％and 39.06 mL,respectively,between the high and low parent strains.The proportions of pedi-grees exceeding the mid-parent value ranged from 29.04％to 77.31％,and the proportions of pedigrees exceeding the high-parent value ranged from 7.08％to 39.56％.The order of variation coefficient was sedimentation value(10.99％)>wet gluten content(6.07％)>crude protein content(5.70％),and the three traits showed extremely significant positive correla-tion(P<0.001).The 551 F6:7 pedigrees were clustered into four groups(Ⅰ～Ⅳ),including 148,125,171 and 107 materi-als,respectively.Group IV displayed the highest average crude protein content(16.87％),wet gluten content(35.96％)and sedimentation value(45.02 mL),making it the superior group.According to the transcriptome sequencing data of high and low value strains screened out 4368 DEGs,in which were 970 upregulated genes and 3398 downregulated genes.GO functional annotation results showed that the differential expression of wheat protein traits mainly occurring in the cellular part,involving membrane structures,organelles,and cell connections,the coordinated variation of serial biological processes occurred by regulating binding and catalytic molecular functions including cellular process,metabolic process,defense response and response to stimulus pathways.KEGG signaling pathway enrichment results showed that the DEGs were significantly(P<0.05)enriched into nine metabolism pathways,four metabolism pathways screened and their enriched DEGs were closely related to wheat grain protein traits,10 DEGs were related to histidine metabolism,21 related to starch and sucrose metabolism,8 related to tyrosine metabolism,8 related to alanine,aspartate and glutamate metabo-lism.In them,there were 7 upregulated expression genes.The results of real-time quantitative PCR verification were basi-cally consistent with the RNA-Seq results.[Conclusion]The grain protein traits belong to quantitative trait inheritance controlled by multiple genes in wheat,and many super-parents types were appeared in F6:7 pedigrees.The variation coeffi-cient of sedimentation value had greater potential for improvement.Group IV was superior group in grain protein traits and could be used in the breeding practices of varieties with superior protein traits.Four pathways were screened,includ-ing histidine metabolism,starch and sucrose metabolism,tyrosine metabolism,and alanine,aspartate and glutamate metabolism based on enrichment functional analysis,were related to the regulation mechanism of wheat grain protein traits.7 candidate genes were identified possibly related to wheat grain protein traits.

关键词

小麦/家系/粗蛋白含量/湿面筋含量/沉降值/转录组

Key words

wheat/pedigree/crude protein content/wet gluten content/sedimentation value/RNA-Seq

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基金项目

国家自然科学基金项目(32160452)

宁夏自然科学基金项目(2023AAC03060)

出版年

2023
南方农业学报
广西壮族自治区农业科学院

南方农业学报

CSTPCDCSCD北大核心
影响因子：0.83
ISSN：2095-1191
参考文献量8
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