抗叶锈病小偃麦代换系WTS135的遗传学分析与分子标记开发

doi:10.11983/CBB25014

摘要/Abstract

摘要： 由于人工驯化与现代育种操作, 普通小麦(Triticum aestivum)的遗传多样性日渐狭窄, 更容易受到病虫害威胁。通过远缘杂交将野生近缘种的抗病基因导入小麦, 有助于拓宽小麦的遗传基础, 为培育抗病品种提供新抗源。十倍体长穗偃麦草(Thinopyrum ponticum)是小麦遗传改良中应用最为广泛的近缘物种之一, 对小麦锈病等多种病害表现出良好抗性。利用远缘杂交和染色体工程, 我们创制了1份小麦-长穗偃麦草种质材料WTS135, 对叶锈菌(Puccinia triticina)生理小种THTT表现免疫。系谱分析表明, 其叶锈病抗性来源于长穗偃麦草外源染色体。顺序基因组原位杂交(GISH)-荧光原位杂交分析显示, 1对十倍体长穗偃麦草染色体替换了小麦7D染色体。液相芯片分析表明外源染色体属于第7部分同源群, 其近着丝粒区的信号密度及丰度明显较低, 与GISH分析结果互相佐证, 因此推测WTS135是1个7St(7D)的二体异代换系。通过分子标记检测, WTS135携带的抗病基因与已知的长穗偃麦草第7部分同源群抗叶锈病基因Lr19和Lr29不同, 推测有可能为1个抗叶锈病新基因。借助Specific-locus amplified fragment sequencing技术, 开发了10个长穗偃麦草特异引物, 可用于快速追踪WTS135中的外源染色质。表型调查显示, WTS135的产量与轮回亲本济麦22无显著性差异, 可直接用于小麦的抗病育种。

关键词: 小麦, 长穂偃麦草, 叶锈病, 远缘杂交, 代换系, 原位杂交, 分子标记

Abstract:

INTRODUCTION: The genetic diversity of common wheat (Triticum aestivum) has decreased sharply due to the artificial domestication and modern breeding operations, making it more vulnerable to the threats from pests and diseases. Leaf rust, caused by the fungal pathogen Puccinia triticina Eriks. (Pt), is a devastating disease in wheat. Over 80 leaf rust resistance (Lr) genes have been formally identified, with nearly half originating from wheat wild relatives. However, the rapid evolution of Pt physiological races has rendered many Lr genes ineffective against prevalent Pt races. Consequently, identifying novel sources of resistance in wild relatives remains an urgent priority for sustainable wheat breeding.

RATIONALE: As one of the most widely used relatives in the genetic improvement of wheat, decaploid Thinopyrum ponticum (Podp.) Barkworth and D. R. Dewey shows excellent resistance to multiple diseases including leaf rust. By wild hybridization and chromosome engineering, we created a wheat-Th. ponticum germplasm WTS135. We evaluated its disease resistance with Pt race THTT, developed Th. ponticum specific markers by specific-locus amplified fragment sequencing technology and assessed its agronomic traits by phenotypic investigation. Sequential genomic in situ hybridization (GISH)-fluorescence in situ hybridization analysis (FISH) and liquid chip have been used to discover its chromosome composition.

RESULTS: WTS135 is immune to the Pt race THTT. Pedigree analysis showed that this resistance originated from the exogenous chromosome of Th. ponticum. GISH-FISH analysis revealed that the wheat chromosomes 7D were replaced by the Th. ponticum-derived chromosomes. Liquid chip showed that the alien chromosomes belonged to the homoeologous group 7, and the density and abundance of the signals in the peri-centromeric region along them were significantly lower, which was consistent with the GISH results. Therefore, it is indicated that WTS135 is a 7St (7D) disomic substitution line. After detected by the molecular markers related to known Lr genes on wheat 7D chromosome, it is presumed that WTS135 could probably carry a novel resistance gene that is not identical to genes Lr19 and Lr29. Ten primers specific to Th. ponticum were developed to rapidly trace the exogenous chromatin in WTS135. Phenotypic investigation showed that the yield of WTS135 was not significantly different from that of the recurrent parent Jimai 22, suggesting that this line can be useful for improving disease resistance in wheat.

CONCLUSION: Introducing resistance genes from wild relatives into wheat through wide hybridization can broaden the genetic base of wheat and provide new sources for breeding disease-resistant varieties. We developed a wheat-Th. ponticum 7St (7D) substitution line, which possibly has a novel alien resistance gene and could be used in wheat disease resistance breeding.

Chromosome composition and leaf rust resistance evaluation of WTS135 (A) GISH analysis using Th. ponticum gDNA as a probe and CS gDNAs as a block; (B) mc-FISH analysis using combined probes (bars=20 μm). (C) The liquid chip analysis of WTS135. (D) WTS135 (1), Jimai 22 (2), Xiaoyan 81 (3), Zhongnong 28 (4), Th. Ponticum (5). The white arrows indicate exogenous chromosomes, purple boxes represent chromosome additions or deletions.

Key words: wheat, Thinopyrum ponticum, leaf rust, wide hybridization, substitution line, in situ hybridization, molecular markers

贾盖亚, 张娜, 李宏伟, 李滨, 李振声, 孔照胜, 郑琪. 抗叶锈病小偃麦代换系WTS135的遗传学分析与分子标记开发. 植物学报, 2025, 60(5): 1-0.

Gaiya Jia, Na Zhang, Hongwei Li, Bin Li, Zhensheng Li, Zhaosheng Kong, Qi Zheng. Genetic Analysis and Molecular Marker Development for a Wheat—Thinopyrum ponticum Substitution Line WTS135 with Leaf Rust Resistance . Chinese Bulletin of Botany, 2025, 60(5): 1-0.

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