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Journal of Integrative Agriculture
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Cytogenetic characterization and molecular marker development of a novel wheat-Thinopyrum ponticum 5E (5D) disomic substitution line with resistance to powdery mildew and stripe rust 
Xiaofang Cheng1, Yi Xiao1, Luhui Wang1, Xiaoying Yang1, Pingchuan Deng1, 2, Jixin Zhao1, 2, Changyou Wang1, 2, Chunhuan Chen1, 2, Tingdong Li1, 2#, Wanquan Ji1, 2#

1 State Key Laboratory of Crop Stress Biology for Arid Areas. College of Agronomy. Northwest A&F University. Yangling 712100. China

2 Shaanxi Research Station of Crop Gene Resources and Germplasm Enhancement, Ministry of Agriculture and Rural Affairs, Yangling 712100. China

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摘要  十倍体长穗偃麦草(2n=10x=70)是普通小麦的野生近缘种之一,因其含有大量抗生物、非生物胁迫的基因,被认为是改良小麦的宝贵遗传资源。本研究以普通小麦7182和十倍体长穗偃麦草杂交的BC1F7衍生系CH97为研究对象。细胞学证据显示,CH97有42条染色体,在减数第一次分裂中期形成21个二价体,随后在减数第一次分裂后期二价体分离并移向两极。通过FISH(荧光原位杂交)、GISH(基因组原位杂交)、mc-GISH(多色基因组原位杂交)和液相芯片综合分析明确了CH97由40条小麦染色体和2条十倍体长穗偃麦草Ee基因组染色体组成,并且缺失了一对小麦的5D染色体,以及1B、6B和7B染色体发生变异。这些结果证实CH97是一个稳定的小麦-十倍体长穗偃麦草5E(5D)二体异代换系。接种试验表明,CH97在整个生育期对小麦白粉病和条锈病表现出高抗,而普通小麦亲本7182呈现高感。与7182相比,CH97的穗粒数、千粒重和粒长均有增加。此外,基于简化基因组测序技术,开发并验证了5E染色体的特异分子标记,开发效率达33.3%,这有助于通过标记辅助选择提高小麦的抗病育种效率。总之,CH97代换系具有的抗病性和改良的农艺性状,为小麦染色体工程育种提供了宝贵的新种质。

Abstract  Thinopyrum ponticum (2n=10x=70), a wild relative of common wheat (Triticum aestivum L.), is considered an invaluable genetic resource for wheat improvement due to its abundance of genes that confer resistance to biotic and abiotic stresses.  This study focused on the CH97 line, derived from the BC1F7 progeny of a cross between wheat cv. 7182 and Th. ponticum.  Cytological evidence showed that CH97 has 42 chromosomes, forming 21 bivalents at meiotic metaphase I, with the bivalents subsequently separating and moving to opposite poles during meiotic anaphase I.  Through a combination of FISH (fluorescence in situ hybridization), GISH (genomic in situ hybridization), mc-GISH (multicolor genomic in situ hybridization), and liquid array analysis, it was determined that CH97 comprises 40 wheat chromosomes and two alien chromosomes from the Ee genome of Th. ponticum, featuring the absence of a pair of 5D chromosomes and variations in 1B, 6B, and 7B chromosomes.  These findings confirm that CH97 is a stable wheat-Th. ponticum 5E (5D) alien disomic substitution line.  Inoculation experiments revealed that CH97 exhibits high resistance to wheat powdery mildew and stripe rust throughout the growth period, in contrast to the highly susceptible common wheat parent 7182.  Compared to 7182, CH97 displayed improvements in spikelets per spike, thousand-kernel weight, and kernel length.  Additionally, utilizing SLAF-seq technology, chromosome 5E-specific molecular markers were developed and validated, achieving a 33.3% success rate, which facilitates marker-assisted selection to enhance disease resistance in wheat.  Overall, the CH97 substitution line, with its resistance to diseases and improved agronomic traits represents valuable new germplasm for wheat chromosome engineering and breeding.
Keywords:  Thinopyrum ponticum        disomic substitution line        powdery mildew        stripe rust SLAF-seq        liquid array  
Online: 10 May 2024  
Fund: This work was funded by the Key R&D project of Yangling Seed Industry Innovation (Ylzy-xm-02) and the Young Elite Scientists Sponsorship Program by CAST (2021QNRC001).
About author:  Xiaofang Cheng, E-mail: chengxf2022@163.com; #Correspondence Wanquan Ji, Tel: 13359215439, E-mail: jiwanquan2008@126.com; Tingdong Li, Tel: 13520437565, E-mail: tingdongli@nwafu.edu.cn.

Cite this article: 

Xiaofang Cheng, Yi Xiao, Luhui Wang, Xiaoying Yang, Pingchuan Deng, Jixin Zhao, Changyou Wang, Chunhuan Chen, Tingdong Li, Wanquan Ji. 2024. Cytogenetic characterization and molecular marker development of a novel wheat-Thinopyrum ponticum 5E (5D) disomic substitution line with resistance to powdery mildew and stripe rust . Journal of Integrative Agriculture, Doi:10.1016/j.jia.2024.04.012

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