Preliminary Mapping of Leaf Rust Resistance Genes in Wheat Line Tian 05156‐1‐1‐2‐1‐2‐1

doi:10.15933/j.cnki.1004-3268.2026.08.003

Abstract

Abstract: According to the previous identification result of resistance to leaf rust at seedling stage，the wheat line Tian 05156‐1‐1‐2‐1‐2‐1 currently exhibited resistance to most physiological races of leaf rust in China.To identify the leaf rust resistance genes carried by this line，Tian 05156‐1‐1‐2‐1‐2‐1 was crossed with the susceptible wheat variety Zhengzhou 5389 to obtain F1 and F2 generations.The leaf rust resistance of the parents and their hybrids was evaluated using THSN，five resistant and five susceptible individual plants were sent to a company for 16K SNP chip analysis，single nucleotide polymorphism（SNP）loci that exhibited polymorphism between the resistant and susceptible small populations were identified.Based on the chromosomes where the differential SNPs were located，KASP markers were designed，KASP markers and known SSR markers were used to detect the parents and F2 populations，and the genotypes of the population were obtained. The leaf rust resistance genes in Tian 05156‐1‐1‐2‐1‐2‐1 were mapped by combining phenotype and genotype data. Candidate genes within the mapping region were identified based on the Chinese Spring reference genome（IWGSC RefSeq V2.1） sequence.The results showed that Tian 05156‐1‐1‐2‐1‐2‐1 exhibited high resistance ［infection type（IT）=1］，while Zhengzhou 5389 demonstrated high susceptibility（IT=4）at seedling stage. All 10 F1 plants displayed resistance（IT≤2）. Among the 200 F₂ individual plants，146 plants were resistant（IT≤2），and 53 plants were susceptible（IT≥3）.Chi‐square test revealed that the number of resistant and susceptible single plants conformed to 3∶1 segregation ratio（χ^²_3∶1=0.284，p>0.05）. It was inferred that Tian 05156‐1‐1‐2‐1‐2‐1 might contain a dominant major gene conferring resistance to leaf rust，and it was designated as LrGW. By using a 16K SNP liquid‐phase chip to detect resistant and susceptible bulks，281 SNPs located on chromosome 1B showed no recombination events in the bulks，indicating that LrGW was likely located on chromosome 1B. Thirty‐one KASP markers were developed based on Chinese Spring reference genome and different SNP loci，and five markers showed good polymorphisms between resistant and susceptible parents，including Zbh1B27，Zbh1B28，Zbh1B30，Zbh1B43，and Zbh1B44. In addition，one known SSR Xbarc8 showed good polymorphism between two parents.Xbarc8 and five KASP markers were used to test the F₂ population，and LrGW was mapped to chromosome 1B between Zbh1B28 and Zbh1B30，with the genetic distances of 0.4 cM and 1.1 cM，respectively，corresponding to the physical interval of 285.7—306.2 Mb in the Chinese Spring reference genome.Analysis of annotated genes within the candidate regions revealed that four kinase genes，including TraesCS1B02G164400，TraesCS1B02G167400，TraesCS1B02G167500，and TraesCS1B02G167700，were potential candidates for LrGW.

Key words: Wheat, Leaf rust, Leaf rust resistance gene, KASP marker, SSR marker, Gene mapping

摘要： 经前期苗期抗叶锈病鉴定，小麦品系天05156-1-1-2-1-2-1对我国多数叶锈菌生理小种表现为抗病。为了确定这个品系所携带的抗病基因，以天05156-1-1-2-1-2-1和感病小麦品种郑州5389杂交获得的F₁和F₂群体为试验材料，用叶锈菌生理小种THSN对双亲及其F₁和F₂进行抗叶锈病鉴定，选择5抗5感单株送公司进行16K SNP芯片分析，挖掘在抗感小群体间表现多态性的SNP（单核苷酸多态性）位点；根据差异SNP所在染色体设计KASP标记，并利用KASP标记和已知SSR标记对亲本及F₂群体进行检测，获得群体基因型；结合表型及基因型对天05156-1-1-2-1-2-1中的抗叶锈病基因进行初步定位；根据中国春参考基因组（IWGSC RefSeq V2.1）序列分析定位区段内的候选基因。结果表明，天05156-1-1-2-1-2-1在苗期表现高抗（侵染型IT=1），郑州5389表现高感（IT=4），10株F₁都表现为抗病（IT≤2），200个F₂单株中鉴定出146个抗病单株（IT≤2）、53个感病单株（IT≥3）。经卡方检测抗感单株数符合3∶1分离比例（χ²_3∶1=0.284，p>0.05），推测天05156-1-1-2-1-2-1中含有1个显性主效抗叶锈病基因，并将其命名为LrGW。通过16K SNP液相芯片检测抗感小群体获得位于1B染色体上的281个SNP位点，其于抗感小群体中表现为0交换，推断该基因可能位于1B染色体上。基于中国春参考基因组并结合差异SNP位点开发了31个KASP标记，5个标记在亲本间表现良好多态性，分别为Zbh1B27、Zbh1B28、Zbh1B30、Zbh1B43、Zbh1B44；此外，已知SSR标记Xbarc8在双亲中也顺利分型。利用Xbarc8和5个KASP标记检测F₂群体，构建遗传连锁图谱，将LrGW基因定位于1B染色体Zbh1B28与Zbh1B30之间，遗传距离分别为0.4 cM和1.1 cM，对应中国春参考基因组物理区间285.7—306.2Mb。对候选区段内的注释基因进行分析，4 个与抗病相关的激酶基因可能是LrGW的候选基因，分别为TraesCS1B02G164400、TraesCS1B02G167400、TraesCS1B02G167500、TraesCS1B02G167700。

关键词: 小麦, 叶锈病, 抗叶锈病基因, KASP标记, SSR标记, 基因定位

CLC Number:

S512

Wang Xinrui, Bai Bin, Wang Jinshuo, Gao Pu, Wang Xinhai, Liu Yiran, Wang Hanbing, Li Zaifeng, Dong Haijiao, Zhang Peipei. Preliminary Mapping of Leaf Rust Resistance Genes in Wheat Line Tian 05156‐1‐1‐2‐1‐2‐1[J]. Journal of Henan Agricultural Sciences, 2026, 55(8): 22-31.

王新锐, 白斌, 王金硕, 高璞, 王新海, 刘祎然, 王涵冰, 李在峰, 董海焦, 张培培. 小麦品系天05156-1-1-2-1-2-1抗叶锈病基因的初步定位[J]. 河南农业科学, 2026, 55(8): 22-31.

Figures/Tables 8

References

［1］海飞，李天杰，郑伟，等. 吡唑醚菌酯与氟环唑复配对小麦叶锈病的防治效果及对小麦的安全性［J］. 农药学学报，2023，25（1）：97‐103.
Hai F，Li T J，Zheng W，et al. Control efficacy of the combination of pyraclostrobin and epoxiconazole against wheat leaf rust and its safety evaluation on wheat plants［J］. Chinese Journal of Pesticide Science，2023，25（1）：97‐103.

［2］Chen X M. Review article：high‐temperature adult‐plant resistance，key for sustainable control of stripe rust［J］.American Journal of Plant Sciences，2013，4（3）：608‐627.

［3］Lin F，Chen X M. Genetics and molecular mapping of genes for race‐specific all‐stage resistance and non‐race‐specific high‐temperature adult‐plant resistance to stripe rust in spring wheat cultivar Alpowa［J］. Theoretical and Applied Genetics，2007，114（7）：1277‐1287.

［4］姚占军，徐世昌，吴立人. 小麦抗病性遗传的研究进展［J］. 辽宁农业科学，2002（6）：30‐33.
Yao Z J，Xu S C，Wu L R. Heredity research progress on disease resistance in wheat［J］. Liaoning Agricultural Sciences，2002（6）：30‐33.

［5］Sharma D，Avni R，Gutierrez‐Gonzalez J，et al. A single NLR gene confers resistance to leaf and stripe rust in wheat［J］. Nature Communications，2024，15：9925.

［6］Zhang P P，Gebrewahid T W，Zhou Y，et al. Seedling and adult plant resistance to leaf rust in 46 Chinese bread wheat landraces and 39 wheat lines with known Lr genes［J］. Journal of Integrative Agriculture，2019，18（5）：1014‐1023.

［7］Wu H，Kang Z H，Li X，et al. Identification of wheat leaf rust resistance genes in Chinese wheat cultivars and the improved germplasms［J］. Plant Disease，2020，104（10）：2669‐2680.

［8］Brevis J C，Chicaiza O，Khan I A，et al. Agronomic and quality evaluation of common wheat near‐isogenic lines carrying the leaf rust resistance gene Lr47［J］. Crop Science，2008，48（4）：1441‐1451.

［9］Tautz D，Trick M，Dover G A. Cryptic simplicity in DNA is a major source of genetic variation［J］. Nature，1986，322（6080）：652‐656.

［10］陈芳，常松鹤，王聪雅，等. 小麦隐性抗白粉病基因pmCH1853的遗传分析与分子定位［J］. 植物保护学报，2025，52（4）：822‐831.
Chen F，Chang S H，Wang C Y，et al. Genetic analysis and molecular mapping of a recessive powdery mildew resistance gene pmCH1853 in wheat［J］. Journal of Plant Protection，2025，52（4）：822‐831.

［11］王俊美，康振生，刘红彦. 小麦农家种红蚰麦抗白粉病遗传分析及SSR分子标记［J］. 植物病理学报，2009，39（3）：285‐289.
Wang J M，Kang Z S，Liu H Y. Genetic analysis and location of a powdery mildew resistance gene in wheat land‐race Hongyoumai by SSR marker［J］. Acta Phytopathologica Sinica，2009，39（3）：285‐289.

［12］李宝，张慧，王同华，等. 高油酸油菜品种‘高沣9号’的选育［J］. 湖南农业科学，2025（10）：1‐6.
Li B，Zhang H，Wang T H，et al. Breeding of the high‐oleic acid variety‘Gaofeng 9’of Brassica napus［J］.Hunan Agricultural Sciences，2025（10）：1‐6.

［13］董亚超. 小麦抗条锈病基因定位与分子标记开发［D］. 乌鲁木齐：新疆农业大学，2024.
Dong Y C. Stripe rust resistance gene mapping and molecular marker development in wheat［D］. Urumqi：Xinjiang Agricultural University，2024.

［14］赵微. 小麦抗秆锈病基因的分子定位及KASP标记开发［D］. 泰安：山东农业大学，2025.
Zhao W. Characterization of stem rust resistance gene and development of KASP markers in wheat［D］.Taian：Shandong Agricultural University，2025.

［15］周悦，李在峰，李星，等. 小麦品系天95HF2抗叶锈基因定位［J］. 作物学报，2010，36（8）：1265‐1269.
Zhou Y，Li Z F，Li X，et al. Molecular mapping of leaf rust resistance genes in wheat line Tian 95HF2［J］.Acta Agronomica Sinica，2010，36（8）：1265‐1269.

［16］王雪晴. 99份小麦材料抗叶锈性鉴定及Ceruga-18中抗叶锈病基因定位［D］. 保定：河北农业大学，2022.
Wang X Q. Identification of leaf rust resistance of 99 wheat materials and mapping of leaf rust resistance genes in Ceruga‐18［D］.Baoding：Hebei Agricultural University，2022.

［17］Roelfs A P，Singh R P，Saari E E. Rust diseases of wheat：concepts and methods of disease management［R］. Mexico：CIMMYT，1992.

［18］张晓祥，王玲，寿路路. 一种快速提取小麦基因组DNA 的改良CTAB方法［J］. 中国农学通报，2012，28（36）：46‐49.
Zhang X X，Wang L，Shou L L. A rapid modified CTAB method of extracting genomic DNA from wheat leaf［J］. Chinese Agricultural Science Bulletin，2012，28（36）：46‐49.

［19］Gao P，Liu P P，Dong R，et al. Identification and molecular marker detection of leaf rust resistance genes in wheat lines from China：development of an efficient software for gene postulation［J］.Phytopathology，2025，115（4）：422‐430.

［20］Somers D J，Isaac P，Edwards K. A high‐density microsatellite consensus map for bread wheat（Triticum aestivum L.）［J］. Theoretical and Applied Genetics，2004，109（6）：1105‐1114.

［21］Kolmer J A，Long D L，Hughes M E. Physiologic specialization of Puccinia triticina on wheat in the United States in 2007［J］. Plant Disease，2009，93（5）：538‐544.

［22］Che M Z，Hiebert C W，McCartney C A，et al. Mapping and DNA marker development for Lr33 from the leaf rust resistant line KU168‐2［J］. Euphytica，2019，215 （2）：29.

［23］Dyck P L. The transfer of leaf rust resistance from Triticum turgidum ssp. dicoccoides to hexaploid wheat［J］.Canadian Journal of Plant Science，1994，74（4）：671‐673.

［24］Singh R P，Mujeeb‐Kazi A，Huerta‐Espino J. Lr46：a gene conferring slow‐rusting resistance to leaf rust in wheat［J］. Phytopathology，1998，88（9）：890‐894.

［25］Helguera M，Vanzetti L，Soria M，et al. PCR markers for Triticum speltoides leaf rust resistance gene Lr51 and their use to develop isogenic hard red spring wheat lines［J］. Crop Science，2005，45（2）：728‐734.

［26］Pietrusińska A，Tyrka M. Linkage of Lr55 wheat leaf rust resistance gene with microsatellite and DArT‐based markers［J］. Physiological and Molecular Plant Pathology，2021，115：101674.

［27］Singh D，Mohler V，Park R F. Discovery，characterisation and mapping of wheat leaf rust resistance gene Lr71［J］.Euphytica，2013，190（1）：131‐136.

［28］Singla J，Lüthi L，Wicker T，et al. Characterization of Lr75：a partial，broad‐spectrum leaf rust resistance gene in wheat［J］. Theoretical and Applied Genetics，2017，130（1）：1‐12.

［29］杨文香，贾璇，闫红飞，等. 小麦抗叶锈基因Lr44的AFLP分子标记［J］. 河北农业大学学报，2005，28（2）：67‐70.
Yang W X，Jia X，Yan H F，et al. Identification of AFLP markers linked to Lr44 resistant to wheat leaf rust［J］.Journal of Agricultural University of Hebei，2005，28（2）：67‐70.

［30］傅向东，刘倩，李振声，等. 小麦基因组研究现状与展望［J］. 中国科学院院刊，2018，33（9）：909‐914.
Fu X D，Liu Q，Li Z S，et al. Research achievement and prospect development on wheat genome［J］.Bulletin of Chinese Academy of Sciences，2018，33（9）：909‐914.

［31］陈中旭. 小麦BSA基因定位方法研究及工具（外显子芯片与WheatGmap）开发［D］. 雅安：四川农业大学，2022.
Chen Z X. Study of wheat BSA gene mapping and tools（exome panel and wheat gmap） development［D］.Yaan：Sichuan Agricultural University，2022.

［32］Krasileva K V，Vasquez‐Gross H A，Howell T，et al.Uncovering hidden variation in polyploid wheat［J］.Proceedings of the National Academy of Sciences of the United States of America，2017，114（6）：913‐921.

［33］刘志杰，王新海，高璞，等. 小麦成株期抗叶锈病基因Lr12精细定位［J］. 华北农学报， 2024，39（5）：179‐185.
Liu Z J，Wang X H，Gao P，et al. Fine mapping of the adult‐plant leaf rust resistance gene Lr12 in wheat［J］.Acta Agriculturae Boreali‐Sinica，2024，39（5）：179‐185.

［34］贺钟锐，徐浩，闫伟宁，等. 小麦抗叶锈基因Lr3ka的定位［J］. 麦类作物学报，2025，45（3）：322‐328.
He Z R，Xu H，Yan W N，et al. Mapping of the leaf rust resistance gene Lr3ka in wheat［J］. Journal of Triticeae Crops，2025，45（3）：322‐328.

［35］徐浩. 意大利小麦品种Lampo中抗叶锈基因LrLa的定位［D］. 保定：河北农业大学，2025.
Xu H. Mapping of the leaf rust resistance gene LrLa in the Italian wheat variety Lampo［D］. Baoding：Hebei Agricultural University，2025.
［36］闫伟宁. 79份小麦品种抗叶锈病种质资源筛选及抗叶锈病基因的发掘［D］. 保定：河北农业大学，2024.
Yan W N. Screening of leaf rust resistance germplasm resources and discovery of leaf rust resistance genes in 79 wheat varieties［D］. Baoding：Hebei Agricultural University，2024.
［37］董海滨，王永霞，方宇辉，等. 黄淮南部小麦重要性状功能基因的KASP标记检测与分析［J］. 江苏农业科学，2025，53（17）：34‐42.

Dong H B，Wang Y X，Fang Y H，et al. KASP marker detection and analysis for functional genes of important wheat traits in southern Huanghuai region［J］. Jiangsu Agricultural Sciences，2025，53（17）:34‐42.
［38］徐晓伟，冯晶，王凤涛，等. 小麦地方品种蚕老麦成株抗条锈病QTL定位［J］. 作物学报，2025，51（11）：2933‐2943.
Xu X W，Feng J，Wang F T，et al. QTL mapping of adult plant resistance to stripe rust in the Chinese wheat landrace Canlaomai ［J］. Acta Agronomica Sinica，2025，51（11）：2933‐2943.
［39］彭律文，雷雨，段继华，等. 基于CsAL1早生关联SNP的KASP分子标记开发与验证［J］. 茶叶通讯，2025，52（4）：446‐452.
Peng L，Lei Y，Duan J H，et al. Development and validation of KASP markers based on the early‐sprouting associated SNP in CsAL1［J］. Journal of Tea Communication，2025，52（4）：446‐452.
［40］Bai B K，Kaur B，Kaur S，et al. Unveiling leaf rust resistance introgressed from non‐progenitor wild species Aegilops kotschyi into hexaploid wheat［J］.Scientific Reports，2025，15：44063.
［41］Kumar S，Bhardwaj S C，Gangwar O P，et al. Lr80：a new and widely effective source of leaf rust resistance of wheat for enhancing diversity of resistance among modern cultivars［J］. Theoretical and Applied Genetics，2021，134（3）：849‐858.

[1]	Zhang Jun, Wang Haijiang, Zhang Lu, Song Peng, Guo Binbin, Zhou Shuang, Ma Chao. Cloning，Bioinformatics and Expression Analysis of TaHsc70‑2 Genes in Wheat [J]. Journal of Henan Agricultural Sciences, 2026, 55(8): 32-41.
[2]	Liu Yuhang, Shi Yuhan, Yang Yiting, Li Xueting, Jiang Mengxin, Li Jiayi, Li Junhua, Sang Shifei, Feng Liuchun, Wang Li. Comprehensive Evaluation of Drought Tolerance and Physiological Regulatory Mechanism of Wheat Seedlings [J]. Journal of Henan Agricultural Sciences, 2026, 55(7): 1-10.
[3]	ZHOU Jinghang, WANG Zenghao, ZHANG Lei, ZHANG Xia, LIU Jiajia, HU Tiezhu, ZHANG Dazhong. Effects of Addition Amounts，Treatment Durations and Pretreatment Durations of Different Chalcogens on Cadmium Absorption and Transport of Wheat Seedlings under Cadmium Stress [J]. Journal of Henan Agricultural Sciences, 2026, 55(6): 1-10.
[4]	WANG Junmei, LIU Lulu, LIU Kai, LI Yahong, FENG Chaohong, SHI Ruijie, HAN Zihang, DU Xijun, XU Fei. Molecular Response of Wheat Serine/Threonine Kinase to Blumeria graminis f.sp.tritici and Plant Hormones [J]. Journal of Henan Agricultural Sciences, 2026, 55(6): 81-91.
[5]	LIANG Lu, YIN Kun, ZHANG Weijie, ZHUANG Qiuli, XU Bohan, TIAN Kaiyue, ZHANG Dongjie, ZHANG Donghui, HUANG Yubo. Impact of Vermicompost Application Rate for Winter Wheat on Dry Matter Accumulation，Transportation，and Yield of Subsequent Maize in Lime Concretion Black Soil [J]. Journal of Henan Agricultural Sciences, 2026, 55(5): 43-52.
[6]	MA Xuhui, ZHANG Fuyan, ZHAO Wan, WANG Jiahuan, CHENG Zhongjie, ZHANG Zhongzhou, LI Liulong, DING Jiacun, ZHAO Chi, WANG Hanfeng, CHEN Xiaojie. Analysis of High and Stable Yield Performance and Genetic Characteristics of New Wheat Cultivar Yutong 112 [J]. Journal of Henan Agricultural Sciences, 2026, 55(4): 23-32.
[7]	ZHANG Yunhong, YANG Yonghui, ZHANG Sensen, ZHANG Zeli, ZHANG Shuiqing, GAO Cuimi, HE Fang, HAN Weifeng, ZHANG Huan, PAN Xiaoying, WANG Bohan. Effects of Different Soil Conditioners on the Quality of Acidified Sandy Ginger Black Soil，and the Growth and Nutrient Absorption of Wheat [J]. Journal of Henan Agricultural Sciences, 2026, 55(4): 79-89.
[8]	TAO Hua, DING Jingli, REN Zhijie, HE Ailing, LIU Gaoyuan, DU Jun. Effects of Nitrogen Reduction Combined with Different Types of Fertilizers on Yield and Nitrogen Use of Winter Wheat [J]. Journal of Henan Agricultural Sciences, 2026, 55(2): 1-11.
[9]	ZHANG Suyu, LIU Hongjie, REN Dechao, YANG Mingda, GE Jun, ZHAO Jingling, ZHU Peipei, ZHENG Dongfang. Effects of Exogenous Regulators on Superior and Inferior Grain Filling and Yield of Wheat [J]. Journal of Henan Agricultural Sciences, 2025, 54(9): 23-33.
[10]	WANG Haodong, LU Yifang, YANG Shan, LIU Fengying, WANG Gang. Effect of the gacS Gene in Pseudomonas chlororaphis on the Biocontrol Efficacy against Wheat Take‑all Disease [J]. Journal of Henan Agricultural Sciences, 2025, 54(9): 112-118.
[11]	MA Shangying, XIA Tingting, HAN Pengbin, ZHANG Mengjiao, MAO Yingjie, WANG Zhiqiang, XIN Zeyu, LIN Tongbao, LIAN Yanhao, REN Yongzhe. Effects of Application Rate and Method of Nitrogen on Winter Wheat Nitrogen Uptake and Utilization，Yield and Soil Nitrate Nitrogen Residue under Wide‑Narrow Row Planting [J]. Journal of Henan Agricultural Sciences, 2025, 54(8): 38-50.
[12]	LIU Xinglin, LIU Yuan, YANG Fan, LIU Buchun, HAN Rui. Research on County‑Level Yield Simulation of Winter Wheat in Henan Province Based on Machine Learning Algorithms [J]. Journal of Henan Agricultural Sciences, 2025, 54(8): 167-180.
[13]	ZANG Hecang, ZHOU Meng, WANG Yahui, PENG Yilong, ZHAO Qing, ZHANG Jie, LI Guoqiang. Study on Wheat Spike Automatic Detection Method Based on Improved YOLOv8n [J]. Journal of Henan Agricultural Sciences, 2025, 54(7): 162-169.
[14]	WANG Haiyang, JIN Haiyang, SONG Hang, PAN Xiuyan, YAN Yaqian, YANG Xiwen, ZENG Zhaohai, ZANG Huadong, ZHENG Nian, LI Xiangdong, HE Dexian. Effects of Different Preceding Crops on Dry Matter Accumulation and Transportation，Nitrogen Absorption and Utilization and Yield of Winter Wheat [J]. Journal of Henan Agricultural Sciences, 2025, 54(6): 1-10.
[15]	LI Xiaode, BAI Xintong, WU Enzhao, LÜ Chunlei, CHEN Shulin, YIN Guihong, WANG Daowen, ZHANG Kunpu. Composition Analysis of Vernalization and Photoperiod Genes in Varieties（lines）Derived from Core Wheat Parent Zhou 8425B [J]. Journal of Henan Agricultural Sciences, 2025, 54(5): 23-30.