摘要
【目的】对宁春4号和河东乌麦杂交、自交F_(6:7)家系进行籽粒蛋白质性状及候选基因分析,以期挖掘与籽粒蛋白质性状相关的基因,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号和河东乌麦)及其杂交、自交的551个F_(6:7)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,通过高、低值株系转录组测序,对差异表达基因(DEGs)进行功能注释及信号通路富集分析,并鉴定籽粒蛋白质性状相关基因。【结果】籽粒蛋白质相关性状在F_(6:7)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%),且三者间呈极显著正相关(P<0.001)。551个F_(6:7)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、胁迫反应和应激反应为主的生物过程协同变化。KEGG信号通路富集结果显示,DEGs显著富集到9条代谢通路(P<0.05),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个与丙氨酸和天冬氨酸及谷氨酸代谢相关,其中有7个DEGs上调表达。实时荧光定量PCR验证结果与转录组测序结果基本一致。【结论】小麦籽粒蛋白质性状属多基因控制的数量性状遗传,F_(6:7)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。
【Objective】Analysis of grain protein traits and candidate genes in F_(6:7) pedigrees of Ningchun No.4 and Hedong black wheat through the cross and self-cross was carried out to discover candidate genes associated with grain protein traits,which would provide theoretical reference for genetic improvement of grain protein traits in wheat.【Method】551 F_(6:7) 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 sedimentation value),transcriptome sequencing,functional annotation of differentially expressed genes(DEGs),signa-ling pathway enrichment analysis were performed for high and low value strains,and genes related to grain protein traits were identified.【Result】The grain protein-related traits in the F_(6:7) 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 values of the traits of both parents.The proportions of pedigrees 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 correlation(P<0.001).The 551 F_(6:7) pedigrees were clustered into four groups(Ⅰ-Ⅳ),including 148,125,171 and 107 materials,respectively.GroupⅣdisplayed 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,4368 DEGs were screened out,in which there were 970 up-regulated genes and 3398 down-regulated genes.GO functional annotation results showed that the differential expression of wheat protein traits mainly occurred in the cellular part,involving cell membrane structure and organelles,but also occurred in cell junctions parts,through regulating molecular functions such as binding and catalysis,it caused synergistic changes in biological processes,mainly cellular processes,metabolic processes,stress responses and stimulation responses.KEGG signaling pathway enrichment results showed that the DEGs were significantly(P<0.05)enriched into nine metabolism pathways,four metabolism pathways were 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 metabolism.In them,there were 7 up-regulated expression genes.The results of real-time fluorescence quantitative PCR verification were basically consistent with the transcriptome sequencing 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 F_(6:7) pedigrees.Sedimentation value had greater potential for improvement.GroupⅣis superior group in grain protein traits and can be used in the breeding practices of varieties with superior protein traits.Four pathways are screened,including histidine metabolism,starch and sucrose metabolism,tyrosine metabolism,and alanine,aspartate and glutamate metabolism based on enrichment functional analysis,are related to the regulation mechanism of wheat grain protein traits.Seven candidate genes are identified possibly related to wheat grain protein traits.
作者
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
CHEN Jia-jing;MAO Xin-yuan;WANG Xin-jie;LI Qing-feng;LIU Cai-xia;ZHANG Xue-ting;ZHAO Bu-quan;KANG Ling;LI Qian-rong;WANG Zhang-jun(Agricultural College,Ningxia University,Yinchuan,Ningxia 750021,China;Institute of Crop Sciences,Ningxia Academy of Agricultural-forestry Sciences,Yinchuan,Ningxia 750001,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2023年第12期3475-3487,共13页
Journal of Southern Agriculture
基金
宁夏自然科学基金项目(2023AAC03060)
宁夏农业育种专项(2023NYYZ02)。
关键词
小麦
家系
粗蛋白含量
湿面筋含量
沉降值
转录组
wheat
pedigree
crude protein content
wet gluten content
sedimentation value
transcriptome
