Kou Cheng, Peng ChaoJun, Dong HaiBin, Hu Lin, Xu WeiGang
College of Agronomy, Northwest A&F University, Xianyang, China.
Institute of Crop Molecular Breeding, Henan Academy of Agricultural Sciences, Zhengzhou, Henan, China.
Front Plant Sci. 2023 Feb 2;14:1118777. doi: 10.3389/fpls.2023.1118777. eCollection 2023.
Pre-harvest Sprouting (PHS) seriously affects wheat quality and yield. However, to date there have been limited reports. It is of great urgency to breed resistance varieties quantitative trait nucleotides (QTNs) or genes for PHS resistance in white-grained wheat.
629 Chinese wheat varieties, including 373 local wheat varieties from 70 years ago and 256 improved wheat varieties were phenotyped for spike sprouting (SS) in two environments and genotyped by wheat 660K microarray. These phenotypes were used to associate with 314,548 SNP markers for identifying QTNs for PHS resistance using several multi-locus genome-wide association study (GWAS) methods. Their candidate genes were verified by RNA-seq, and the validated candidate genes were further exploited in wheat breeding.
As a result, variation coefficients of 50% and 47% for PHS in 629 wheat varieties, respectively, in 2020-2021 and 2021-2022 indicated large phenotypic variation, in particular, 38 white grain varieties appeared at least medium resistance, such as Baipimai, Fengchan 3, and Jimai 20. In GWAS, 22 significant QTNs, with the sizes of 0.06% ~ 38.11%, for PHS resistance were stably identified by multiple multi-locus methods in two environments, e.g., AX-95124645 (chr3D:571.35Mb), with the sizes of 36.390% and 45.850% in 2020-2021 and 2021-2022, respectively, was detected by several multi-locus methods in two environments. As compared with previous studies, the AX-95124645 was used to develop Kompetitive Allele-Specific PCR marker QSS.TAF9-3D (chr3D:569.17Mb~573.55Mb) for the first time, especially, it is available in white-grain wheat varieties. Around this locus, nine genes were significantly differentially expressed, and two of them (TraesCS3D01G466100 and TraesCS3D01G468500) were found by GO annotation to be related to PHS resistance and determined as candidate genes.
The QTN and two new candidate genes related to PHS resistance were identified in this study. The QTN can be used to effectively identify the PHS resistance materials, especially, all the white-grained varieties with QSS.TAF9-3D-TT haplotype are resistant to spike sprouting. Thus, this study provides candidate genes, materials, and methodological basis for breeding wheat PHS resistance in the future.
收获前发芽(PHS)严重影响小麦品质和产量。然而,迄今为止相关报道有限。培育白粒小麦中抗收获前发芽的数量性状核苷酸(QTNs)或基因迫在眉睫。
629个中国小麦品种,包括70年前的373个地方小麦品种和256个改良小麦品种,在两种环境下对穗发芽(SS)进行表型鉴定,并通过小麦660K芯片进行基因分型。利用这些表型与314,548个单核苷酸多态性(SNP)标记进行关联分析,采用多种多位点全基因组关联研究(GWAS)方法鉴定抗收获前发芽的QTNs。通过RNA测序验证其候选基因,并在小麦育种中进一步利用经过验证的候选基因。
结果表明,629个小麦品种在2020 - 2021年和2021 - 2022年收获前发芽的变异系数分别为50%和47%,表明表型变异较大,特别是有38个白粒品种表现出至少中等抗性,如白麦、丰产3号和济麦20。在GWAS分析中,通过多种多位点方法在两种环境下稳定鉴定出22个与抗收获前发芽相关的显著QTNs,其贡献率为0.06%至38.11%,例如AX - 95124645(chr3D:571.35Mb),在2020 - 2021年和2021 - 2022年的贡献率分别为36.390%和45.850%,在两种环境下均被多种多位点方法检测到。与以往研究相比,首次利用AX - 95124645开发了竞争性等位基因特异性PCR标记QSS.TAF9 - 3D(chr3D:569.17Mb~573.55Mb),特别是该标记在白粒小麦品种中可用。在该位点周围,有9个基因显著差异表达,通过基因本体(GO)注释发现其中两个基因(TraesCS3D01G466100和TraesCS3D01G468500)与抗收获前发芽相关,并确定为候选基因。
本研究鉴定了与抗收获前发芽相关的QTN和两个新的候选基因。该QTN可有效鉴定抗收获前发芽材料,特别是所有具有QSS.TAF9 - 3D - TT单倍型的白粒品种均抗穗发芽。因此,本研究为未来小麦抗收获前发芽育种提供了候选基因、材料和方法学基础。