Comparative molecular cytogenetic analysis and molecular marker development of wheat–Leymus mollis and wheat-Psathyrostachys huashanica 7Ns alien chromosome lines

doi:10.1016/j.jia.2026.01.028

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Comparative molecular cytogenetic analysis and molecular marker development of wheat–Leymus mollis and wheat-Psathyrostachys huashanica 7Ns alien chromosome lines

Xin Du^1,², Yanlong Jin³, Xiaomin Xu², Yanzhen Wang⁴, Tingting Li⁵, Jixin Zhao², Changyou Wang², Tingdong Li², Chunhuan Chen², Xinlun Liu², Yingxiu Wan¹^#, Pingchuan Deng²^#, Wanquan Ji²^#

¹Institute of Crop Research, Anhui Academy of Agricultural Science, Hefei 230000, China

²State Key Laboratory of Crop Stress Biology in AridAreas, College of Agronomy, Northwest A&F University, Yangling 712100, China

³College of Agriculture, Shihezi University, Shihezi 832000, China

⁴Center for Agricultural Genetic Resources Research, Shanxi Agricultural University, Taiyuan 030031, China

⁵College of Bioengineering, Yangling Vocational Technical College, Yangling 712100, China

Highlights

· The 7Ns chromosomes from Leymus mollis and Psathyrostachys huashanica show distinct structures and confer different spectra of stripe rust resistance.

· A set of 36 molecular markers was developed to precisely track the 7Ns chromosomes in wheat.

· The 7Ns chromosomes from both species provide effective resistance to Fusarium head blight with minimal negative agronomic impacts.

Abstract
References

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摘要

滨麦（Leymus mollis (Trin.) Pilger，NsNsXmXm，2n=28）是小麦族中的一个四倍体多年生物种，因其具有多种优良性状而成为小麦改良的重要三级基因库。本研究对三个小麦外源染色体材料的7Ns染色体进行了系统的细胞遗传学与分子比较，包括小麦滨麦7Ns（7D）二体代换系M10、小麦滨麦7Ns二体附加系M8以及小麦华山新麦草（Psathyrostachys huashanica）7Ns二体附加系H1。结果显示，滨麦与华山新麦草的7Ns染色体在FISH信号分布和臂比上存在明显差异。利用小麦华山新麦草45K液相芯片（GenoBaits®WheatplusPh）进行分析，发现H1中7Ns染色体长臂存在一大段末端缺失，而M10和M8中的7Ns染色体结构完整。三个7Ns材料均表现出对赤霉病的抗性，但对条锈病的反应有所不同：M10和M8在苗期和成株期均对6个主流小种具有广谱抗性，而H1仅对CYR23、CYR31和CYR32免疫。农艺性状评价表明，各材料在株高、穗型、穗长、籽粒大小和粒色等方面存在显著差异，而每穗小穗数、千粒重和芒性等性状无显著变化。所有材料的千粒重均较亲本7182有所提高。基于转录组数据，从7Ns染色体中鉴定出668条染色体特异性unigene，并开发验证了36个染色体特异性分子标记。其中，7Ns染色体共有标记为24个，滨麦7Ns染色体特异标记9个，华山新麦草7Ns染色体特异标记3个。本研究为理解7Ns染色体的分化提供了新见解，并为加速小麦遗传改良提供了实用的分子工具。

Abstract

Leymus mollis (Trin.) Pilger (NsNsXmXm, 2n=28) is a tetraploid perennial species within the Triticeae tribe and constitutes a valuable tertiary gene pool for wheat improvement because of its diverse beneficial traits. In this study, a comprehensive cytogenetic and molecular comparison was performed on the 7Ns chromosomes of three wheat-alien lines: wheat-L. mollis 7Ns (7D) disomic substitution line (M10), the wheat-L. mollis 7Ns disomic addition line (M8), and wheat-Psathyrostachys huashanica 7Ns disomic addition line (H1). Distinct differences in FISH signal distribution and arm ratios were observed between the 7Ns chromosomes of L. mollis and P. huashanica. Analysis using the wheat-P. huashanica 45K liquid array (GenoBaits^®WheatplusPh) revealed a large terminal deletion on the long arm of 7Ns in H1, while the 7Ns chromosomes in M10 and M8 remained structurally intact. All three 7Ns lines exhibited resistance to Fusarium head blight (FHB). However, their responses to stripe rust differed. M10 and M8 displayed broad-spectrum resistance to six predominant races at both seedling and adult stages, whereas H1 was immune only to CYR23, CYR31, and CYR32. The evaluation of agronomic traits identified significant differences among the lines in plant height, spike type, spike length, grain size, and grain color, whereas no significant variation was detected in spikelets per spike, thousand-kernel weight, or awn characteristics. All lines showed an increased thousand-kernel weight compared with the parental line 7182. Based on transcriptome data, 668 chromosome-specific unigenes were identified from 7Ns chromosomes, and 36 chromosome-specific markers were developed and validated. Among these, 24 were common to all 7Ns chromosomes, 9 were specific to L. mollis 7Ns chromosomes, and 3 were specific to P. huashanica 7Ns chromosomes. This work provides new insights into the divergence of 7Ns chromosomes and delivers practical molecular tools to accelerate wheat improvement.

Keywords: Leymus mollis Psathyrostachys huashanica stripe rust and Fusarium head blight 7Ns-specific markers

Online: 20 January 2026

Fund:

This research was supported by National Key R&D Program of China (2023YFD1200404), the Talent Program of Anhui Academy of Agricultural Sciences, China (XXBS-202312), the First Level Youth Talent Program of Anhui Academy of Agricultural Sciences, China (2022), the joint research of improved wheat variety of Anhui, China (2021-), and the China Agriculture Research System of MOF and MARA (CARS-03-78).

About author: #Correspondence Wanquan Ji, E-mail: jiwanquan2008@126.com; Pingchuan Deng, E-mail: dengpingchuan@nwafu.edu.cn; Yingxiu Wan, E-mail: wanyingxiu@163.com

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Xin Du, Yanlong Jin, Xiaomin Xu, Yanzhen Wang, Tingting Li, Jixin Zhao, Changyou Wang, Tingdong Li, Chunhuan Chen, Xinlun Liu, Yingxiu Wan, Pingchuan Deng, Wanquan Ji. 2026. Comparative molecular cytogenetic analysis and molecular marker development of wheat–Leymus mollis and wheat-Psathyrostachys huashanica 7Ns alien chromosome lines. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.01.028

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