Welcome to Acta Agronomica Sinica，

Acta Agronomica Sinica

   

Mapping and identification of a novel sharp eyespot resistance locus Qse.hnau-5AS and its candidate genes in wheat

GAO Meng-Juan,ZHAO He-Ying,CHEN Jia-Hui,CHEN Xiao-Qian,NIU Meng-Kang,QIAN Qi-Run,CUI Lu-Fei,XING Jiang-Min,YIN Qing-Miao,GUO Wen,ZHANG Ning SUN Cong-Wei,YANG Xia,PEI Dan,JIA Ao-Lin,CHEN Feng,YU Xiao-Dong*,REN Yan*   

  1. College of Agronomy, Henan Agricultural University / State Key Laboratory of High-Efficiency Production of Wheat-Maize Double Cropping, Zhengzhou 450046, Henan, China
  • Received:2025-01-17 Revised:2025-04-27 Accepted:2025-04-27 Published:2025-05-13
  • Supported by:
    This study was supported by the National Key Research and Development Program (2022YFD1201504), the Henan Major Science and Technology Project (201100110100), the Science and Technology R & D Plan Joint Fund of Henan Province (242103810020), and the China Postdoctoral Science Foundation (2023M741068).

PDF

7

Abstract:

Sharp eyespot, caused by Rhizoctonia cerealis, is a destructive soil-borne disease that poses a serious threat to wheat production in China, significantly affecting yield stability and productivity. Breeding and deploying resistant varieties is one of the most economical, effective, and environmentally sustainable strategies for disease control. Identifying resistance genes is fundamental to the development of superior resistant varieties. In this study, 349 wheat varieties (or lines) from the Huang-Huai region of China were collected and evaluated for sharp eyespot resistance in an artificial climate chamber at the Wheat Molecular Breeding Innovation Center, Henan Agricultural University. Genotyping was performed using the wheat 660K SNP array. A genome-wide association study (GWAS) was conducted using a mixed linear model (MLM) approach, integrating phenotypic data to identify loci associated with resistance. A novel quantitative trait locus (QTL), designated Qse.hnau-5AS, was identified on the short arm of chromosome 5A. GWAS results revealed 15 significant SNPs clustered within a 960.6 kb genomic region. Haplotype analysis confirmed that this locus significantly enhances resistance to sharp eyespot. Within the Qse.hnau-5AS region, 13 high-confidence annotated genes were identified. Based on expression profiling and response to R. cerealis infection, two candidate genes were proposed: one encoding a Hedgehog-interacting-like protein (TaHIPL) and the other encoding a plasma membrane ATPase (TaHA). Functional validation using virus-induced gene silencing (VIGS) showed that silencing of TaHIPL and TaHA resulted in significant downregulation of gene expression (confirmed by qRT-PCR) and a marked increase in disease index (DI) compared to control plants. These findings indicate that TaHIPL and TaHA positively regulate resistance to sharp eyespot in wheat. This study provides valuable genetic resources for understanding the molecular mechanisms underlying sharp eyespot resistance and for advancing resistance breeding in wheat.

Key words: wheat, sharp eyespot, seedling-stage resistance, GWAS, VIGS

[1] Wu X J, Wang J C, Wu D, Jiang W, Gao Z F, Li D S, Wu R L, Gao D R, Zhang Y. Identification of new resistance loci against wheat sharp eyespot through genome-wide association study. Front Plant Sci, 2022, 13: 1056935.

[2] Chen J, Li G H, Du Z Y, Quan W, Zhang H Y, Che M Z, Wang Z, Zhang Z J. Mapping of QTL conferring resistance to sharp eyespot (Rhizoctonia cerealis) in bread wheat at the adult plant growth stage. Theor Appl Genet, 2013, 126: 2865–2878.

[3] 2024年小麦春季重大病虫害防控技术方案. [2024.12.8]. https://www.moa.gov.cn/gk/nszd_1/nszd_2/202403/t20240308_6450964.htm.

Technical plan for the prevention and control of major diseases and pests of wheat in spring 2024. [2024.12.8]. https://www.moa.gov.cn/gk/nszd_1/nszd_2/202403/t20240308_6450964.htm (in Chinese).

[4] Liu C Y, Guo W, Zhang Q F, Fu B S, Yang Z J, Sukumaran S, Cai J, Liu Y, Zhai W L, Wu X Y, et al. Genetic dissection of adult plant resistance to sharp eyespot using an updated genetic map of Niavt14 × Xuzhou25 winter wheat recombinant inbred line population. Plant Dis, 2021, 105: 997–1005.

[5] Guo Y, Du Z Y, Chen J, Zhang Z J. QTL mapping of wheat plant architectural characteristics and their genetic relationship with seven QTLs conferring resistance to sheath blight. PLoS One, 2017, 12: e0174939.

[6] Wu X J, Cheng K, Zhao R H, Zang S J, Bie T D, Jiang Z N, Wu R L, Gao D R, Zhang B Q. Quantitative trait loci responsible for sharp eyespot resistance in common wheat CI12633. Sci Rep, 2017, 7: 11799.

[7] Jiang Y J, Zhu F F, Cai S B, Wu J Z, Zhang Q F. Quantitative trait loci for resistance to Sharp Eyespot (Rhizoctonia cerealis) in recombinant inbred wheat lines from the cross Niavt 14 × Xuzhou 25. Czech J Genet Plant Breed, 2016, 52: 139–144.

[8] Su J, Zhao J J, Zhao S Q, Li M Y, Pang S Y, Kang Z S, Zhen W C, Chen S S, Chen F, Wang X D. Genetics of resistance to common root rot (spot blotch), Fusarium crown rot, and sharp eyespot in wheat. Front Genet, 2021, 12: 699342.

[9] Ma J H, Ye M M, Liu Q Q, Yuan M, Zhang D J, Li C X, Zeng Q D, Wu J H, Han D J, Jiang L N. Genome-wide association study for grain zinc concentration in bread wheat (Triticum aestivum L.). Front Plant Sci, 2023, 14: 1169858.

[10] Yao F J, Guan F N, Duan L Y, Long L, Tang H, Jiang Y F, Li H, Jiang Q T, Wang J R, Qi P F, et al. Genome-wide association analysis of stable stripe rust resistance loci in a Chinese wheat Landrace panel using the 660K SNP array. Front Plant Sci, 2021, 12: 783830.

[11] Liu S Y, Wang C Y, Gou J Y, Dong Y, Tian W F, Fu L P, Xiao Y G, Luo X M, He Z H, Xia X C, et al. Genome-wide association study of ferulic acid content using 90K and 660K SNP chips in wheat. J Cereal Sci, 2022, 106: 103498.

[12] Wu J H, Wang X T, Chen N, Yu R, Yu S Z, Wang Q L, Huang S, Wang H Y, Singh R P, Bhavani S, et al. Association analysis identifies new loci for resistance to Chinese Yr26-virulent races of the stripe rust pathogen in a diverse panel of wheat germplasm. Plant Dis, 2020, 104: 1751–1762.

[13] 余蓬勃. 小麦纹枯病苗期抗性鉴定方法的改良及全基因组关联分析. 河南农业大学硕士学位论文, 河南郑州, 2019.
Yu P B. Improvement of Identification Method for Resistance to Wheat Sharp Eyespot and Genome-wide Association Analysis. MS Thesis of Henan Agricultural University, Zhengzhou, Henan, China, 2019 (in Chinese with English abstract).

[14] Fernandez-Pozo N, Rosli H G, Martin G B, Mueller L A. The SGN VIGS tool: user-friendly software to design virus-induced gene silencing (VIGS) constructs for functional genomics. Mol Plant, 2015, 8: 486–488.

[15] 于寒松, 彭帅, 谢远红, 胡耀辉. 一种RNA提取试剂盒: TRIZOL的使用方法初探. 食品科学, 2005, 26(11): 39–42.
Yu H S, Peng S, Xie Y H, Hu Y H. Study on improvement of RNA isolating reagent kit: TRIZOL. Food Sci, 2005, 26(11): 39–42 (in Chinese with English abstract).

[16] 董恩妮, 梁青, 李利, 王林杰, 仲涛, 王永, 张红平. 实时荧光定量PCR内参基因的选择. 中国畜牧杂志, 2013, 49(11): 92–96.
Dong E N, Liang Q, Li L, Wang L J, Zhong T, Wang Y, Zhang H P. The selection of reference gene in real-time quantitative reverse transcription PCR. Chin J Anim Sci, 2013, 49(11): 92–96 (in Chinese).

[17] 韩天龙, 王敏, 李志明. QRT-PCR检测目的基因mRNA转录水平的应用. 安徽农业科学, 2011, 39: 18432–18434.
Han T L, Wang M, Li Z M. Application of QRT-PCR in the detection of mRNA transcription level. J Anhui Agric Sci, 2011, 39: 18432–18434 (in Chinese with English abstract).

[18] 吴凯朝, 黄诚梅, 李杨瑞, 杨丽涛, 吴建明. Trizol试剂法快速高效提取3种作物不同组织总RNA. 南方农业学报, 2012, 43: 1934–1939.
Wu K C, Huang C M, Li Y R, Yang L T, Wu J M. Fast and effective total RNA extraction from different tissues in 3 crops through the Trizol reagent method. J South Agric, 2012, 43: 1934–1939 (in Chinese with English abstract).

[19] McBeath J H, McBeath J. Environmental Change and Food Security in China. Duxford: Woodhead Publishing, 2010. pp 119–120.

[20] Ren Y, Yu P B, Wang Y, Hou W X, Yang X, Fan J L, Wu X H, Lv X L, Zhang N, Zhao L, et al. Development of a rapid approach for detecting sharp eyespot resistance in seedling-stage wheat and its application in Chinese wheat cultivars. Plant Dis, 2020, 104: 1662–1667.

[21] 任丽娟, 蔡士宾, 汤颋, 吴纪中, 周淼平, 颜伟, 马鸿翔, 陆维忠. 小麦纹枯病抗性QTL的SSR标记研究. 扬州大学学报(农业与生命科学版), 2004, 25(4): 16–19.
Ren L J, Cai S B, Tang T, Wu J Z, Zhou M P, Yan W, Ma H X, Lu W Z. SSR markers linked resistance QTLs to sharp eyespot (Rhizoctonia cerealis) in wheat. J Yangzhou Univ (Agric Life Sci Edn), 2004, 25(4): 16–19 (in Chinese with English abstract).

[22] 刘颖, 张巧凤, 付必胜, 蔡士宾, 蒋彦婕, 张志良, 邓渊钰, 吴纪中, 戴廷波. 小麦纹枯病抗源的遗传多样性及抗性基因位点SSR标记分析. 作物学报, 2015, 41: 1671–1681.

Liu Y, Zhang Q F, Fu B S, Cai S B, Jiang Y J, Zhang Z L, Deng Y Y, Wu J Z, Dai T B. Genetic diversity of wheat germplasm resistant to sharp eyespot and geno-typing of resistance loci using SSR markers. Acta Agron Sin, 2015, 41: 1671–1681 (in Chinese with English abstract).

[23] Sun C W, Dong Z D, Zhao L, Ren Y, Zhang N, Chen F. The Wheat 660K SNP array demonstrates great potential for marker-assisted selection in polyploid wheat. Plant Biotechnol J, 2020, 18: 1354–1360.

[24] Yang X, Pan Y B, Singh P K, He X Y, Ren Y, Zhao L, Zhang N, Cheng S H, Chen F. Investigation and genome-wide association study for Fusarium crown rot resistance in Chinese common wheat. BMC Plant Biol, 2019, 19: 153.

[25] Liu J, Cobertera D C, Zemetra R S, Mundt C C. Identification of quantitative trait loci for resistance to wheat sharp eyespot in the Einstein × tubbs recombinant inbred line population. Plant Dis, 2023, 107: 820–825.

[26] 李於亭, 熊宏春, 郭会君, 赵林姝, 谢永盾, 古佳玉, 李慧园, 赵世荣, 丁玉萍, 方正武, 等.小麦抽穗期微效QTL精细定位与候选基因分析. 第二十届中国作物学会学术年会. 2023. 
Li Y T, Xiong H C, Guo H J, Zhao L S, Xie Y D, Gu J Y, Li H Y, Zhao S R, Ding Y P, Fang Z W, et al. Fine mapping of micro-effect QTLs at heading stage and analysis of candidate genes in wheat. The 20th Annual meeting of the Crop Science Society of China. 2023 (in Chinese).

[27] 禹海龙, 吴文雪, 裴星旭, 刘晓宇, 邓跟望, 李西臣, 甄士聪, 望俊森, 赵永涛, 许海霞, 等. 小麦茎秆性状的转录组测序及全基因组关联分析. 作物学报, 2024, 50: 2187–2206.
Yu H L, Wu W X, Pei X X, Liu X Y, Deng G W, Li X C, Zhen S C, Wang J S, Zhao Y T, Xu H X, et al. Transcriptome sequencing and genome-wide association study of wheat stem traits. Acta Agron Sin, 2024, 50: 2187–2206 (in Chinese with English abstract).

[28] Li M T, Guo P, Nan N, Ma A, Liu W X, Wang T J, Yun D J, Xu Z Y. Plasma membrane-localized H+-ATPase OsAHA3 functions in saline-alkaline stress tolerance in rice. Plant Cell Rep, 2023, 43: 9.

[29] Kim C Y, Park J Y, Choi G, Kim S, Vo K T X, Jeon J S, Kang S, Lee Y H. A rice gene encoding glycosyl hydrolase plays contrasting roles in immunity depending on the type of pathogens. Mol Plant Pathol, 2022, 23: 400–416.

[1] MENG Xiang-Yu, DIAO Deng-Chao, LIU Ya-Rui, LI Yun-Li, SUN Yu-Chen, WU Wei, ZHAO Wen, WANG Yu, WU Jian-Hui, LI Chun-Lian, ZENG Qing-Dong, HAN De-Jun, ZHENG Wei-Jun. Genetic analysis of high yield and yield stability characteristics of new wheat variety Xinong 877 [J]. Acta Agronomica Sinica, 2025, 51(5): 1261-1276.
[2] WANG Qing, WANG Yi-Xiu, LI Yue-Nan, LYU Yong-Hui, ZHANG Hai-Bo, LIU Na, CHENG Hong-Yan. Differences in transcriptomic responses to cadmium stress in high/low-Cd- accumulation wheat [J]. Acta Agronomica Sinica, 2025, 51(5): 1230-1247.
[3] WANG Jia-Jie, WANG Zheng-Nan, BATOOL Maria, WANG Wang-Nian, WEN Jing, REN Chang-Zhong, HE Feng, WU You-You, XU Zheng-Hua, WANG Jing, KUAI Jie, WANG Bo, ZHOU Guang-Sheng, FU Ting-Dong. Comparison of physiological characteristics of salt and alkali tolerance between rapeseed and wheat [J]. Acta Agronomica Sinica, 2025, 51(5): 1215-1229.
[4] WANG Dong, WANG Sen, SHANG Li, FENG Hao-Wei, ZHANG Yong-Qiao, CUI Jia-Ming, LI Shuang, ZHANG Jia-Cong, CHE Huan. Effect of supplementary irrigation on winter wheat yield and water use efficiency in semi humid areas of the Loess Plateau [J]. Acta Agronomica Sinica, 2025, 51(5): 1312-1325.
[5] CHENG Hong-Na, QIN Dan-Dan, XU Fu-Chao, XU Qing, PENG Yan-Chun, SUN Long-Qing, XU Le, GUO Ying, YANG Xin-Quan, XU De-Ze, DONG Jing. Comparative analysis of metabolomics of colored hulless barley and colored wheat grains [J]. Acta Agronomica Sinica, 2025, 51(4): 932-942.
[6] WANG Lin, CHEN Xiao-Yu, ZHANG Wen-Meng-Long, WANG Si-Qi, CHENG Bing-Yun, CHENG Jing-Qiu, PAN Rui, ZHANG Wen-Ying. Molecular characteristics and functional analysis of HvMYB2 in response to drought stress in barley [J]. Acta Agronomica Sinica, 2025, 51(4): 873-887.
[7] LI Hui-Min, XING Zhi-Peng, ZHANG Hai-Peng, WEI Hai-Yan, ZHANG Hong-Cheng, LI Guang-Yan. Application of chemical regulators and other cultivation measures in lodging resistance and high-yield cultivation of wheat [J]. Acta Agronomica Sinica, 2025, 51(4): 847-862.
[8] LI Pei-Hua, LI Jie, MENG Xiang-Yu, SUN Yu-Chen, FENG Yong-Jia, LI Yun-Li, DIAO Deng-Chao, ZHAO Wen, WU Wei, HAN De-Jun, ZHANG Song-Wu, ZHENG Wei-Jun. Evaluation of stress tolerance and physiological response of cold-type wheat under heat stress [J]. Acta Agronomica Sinica, 2025, 51(4): 1118-1130.
[9] LI Qiao, YE Yang-Chun, CHANG Xu-Hong, WANG De-Mei, WANG Yan-Jie, YANG Yu-Shuang, MA Rui-Qi, ZHAO Guang-Cai, CAI Rui-Guo, ZHANG Min, LIU Xi-Wei. Effects of high temperature and drought stresses on photosynthetic characteristics and yield of winter wheat after anthesis [J]. Acta Agronomica Sinica, 2025, 51(4): 1077-1090.
[10] WANG Jiao, BAI Hai-Xia, HAN Yu-Yan, LIANG Hui, FENG Ya-Nan, ZHANG Dong-Sheng, LI Ping, ZONG Yu-Zheng, SHI Xin-Rui, HAO Xing-Yu. Effects of elevated CO2 concentration, increased temperature and their interaction on the carbon and nitrogen metabolism in Liangxing 99 winter wheat leaves [J]. Acta Agronomica Sinica, 2025, 51(4): 1061-1076.
[11] ZHANG Heng, FENG Ya-Lan, TIAN Wen-Zhong, GUO Bin-Bin, ZHANG Jun, MA Chao. Identification of TaSnRK gene family and expression analysis under localized root zone drought in wheat [J]. Acta Agronomica Sinica, 2025, 51(3): 632-649.
[12] ZHAN Zong-Bing, JIN Qi-Feng, LIU Di, LYU Ying-Chun, GUO Ying, ZHANG Xue-Ting, HU Meng-Xia, WANG Shang, YANG Fang-Ping. Molecular characterization and evaluation of important traits of landrace wheat Laomangmai in Gansu province, China [J]. Acta Agronomica Sinica, 2025, 51(3): 609-620.
[13] XU Jian-Xia, DING Yan-Qing, CAO Ning, CHENG Bin, GAO Xu, LI Wen-Zhen, ZHANG Li-Yi. Genome-wide association analysis and prediction of candidate genes for plant height and internode number in Chinese sorghum [J]. Acta Agronomica Sinica, 2025, 51(3): 568-585.
[14] YANG Fang-Ping, GUO Ying, TIAN Yuan-Yuan, XU Yu-Feng, WANG Lan-Lan, BAI Bin, ZHAN Zong-Bing, ZHANG Xue-Ting, XU Yin-Ping, LIU Jin-Dong. Effect of vernalization and photoperiod genes and evaluation of cold tolerance for wheat landraces from Gansu province, China [J]. Acta Agronomica Sinica, 2025, 51(2): 370-382.
[15] LIANG Miao, LI Pan, ZHAO Lian-Hao, FAN Zhi-Long, HU Fa-Long, FAN Hong, HE Wei, CHAI Qiang, YIN Wen. Effect of soil conditioner and slow-release nitrogen fertilizer on dry matter accumulation and yield of wheat [J]. Acta Agronomica Sinica, 2025, 51(2): 470-484.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd