卵穗山羊草染色体核型构建和基于SLAF-seq特异性分子标记开发

doi:10.1016/j.jia.2023.09.014

摘要/Abstract

摘要：

病原菌的不断进化对小麦的抗病能力构成威胁，危及粮食安全。因此，培育抗病小麦品种是规避这一问题最经济有效的途径。卵穗山羊草SY159是小麦的近缘种，包含有许多优异的基因资源。在这项研究中，我们利用寡核苷酸探针Oligo-pTa535和Oligo-pSc119.2，参考了一整套小麦-卵穗山羊草（TA2899）附加系的荧光原位杂交（FISH）结果，构建了SY159染色体核型。利用SLAF-seq技术，开发了400对特异标记对SY159染色体进行筛选，检测效率达81.5%。同时，以一整套小麦-卵穗山羊草（TA2899）附加系为参考，对特异性标记进行同源群归属的分类，并以源于SY159的7M^g附加系进行验证，证实了利用该技术开发卵穗山羊草SY159不同染色体特异标记的适用性。这为加快卵穗山羊草在小麦遗传育种中的应用奠定了坚实的基础。

Abstract:

The constant evolution of pathogens poses a threat to wheat resistance against diseases, endangering food security. Developing resistant wheat varieties is the most practical approach for circumventing this problem. As a close relative of wheat, Aegilops geniculata, particularly accession SY159, has evolved numerous beneficial traits that could be applied to improve wheat. In this study, we established the karyotype of SY159 by fluorescence in situ hybridization (FISH) using the oligonucleotide probes Oligo-pTa535 and Oligo-pSc119.2 and a complete set of wheat–Ae. geniculata accession TA2899 addition lines as a reference. Using specific-locus amplified fragment sequencing (SLAF-seq) technology, 400 specific markers were established for detecting the SY159 chromosomes with efficiencies reaching 81.5%. The SY159-specific markers were used to classify the different homologous groups of SY159 against the wheat–Ae. geniculata addition lines. We used these specific markers on the 7M^g chromosome after classification, and successfully confirmed their suitability for studying the different chromosomes of SY159. This study provides a foundation for accelerating the application of SY159 in genetic breeding programs designed to improve wheat.

Key words: Aegilops geniculata Roth , chromosome karyotype analysis , FISH , SLAF-seq , specific molecular marker

Yongfu Wang, Jianzhong Fan, Hong Zhang, Pingchuan Deng, Tingdong Li, Chunhuan Chen, Wanquan Ji, Yajuan Wang. 卵穗山羊草染色体核型构建和基于SLAF-seq特异性分子标记开发[J]. Journal of Integrative Agriculture, 2024, 23(12): 3953-3965.

Yongfu Wang, Jianzhong Fan, Hong Zhang, Pingchuan Deng, Tingdong Li, Chunhuan Chen, Wanquan Ji, Yajuan Wang. Karyotype establishment and development of specific molecular markers of Aegilops geniculata Roth based on SLAF-seq [J]. Journal of Integrative Agriculture, 2024, 23(12): 3953-3965.

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