Molecular Mapping of Adult Plant Resistance Gene to Powdery Mildew in Tetraploid Wheat

doi:10.7668/hbnxb.20195107

Abstract

Abstract:

Abstract: Tetraploid wheat is the ancestor specie of common wheat and an important food crop.Aiming to provide new resistance sources for wheat variety breeding,the resistance tetraploid wheat germplasm was explored and their resistance genes were identified.TDI-1 is a cultivated emmer wheat that has been immune to powdery mildew in the field for many years.To determine the resistance genes carried by TDI-1,and provide a theoretical basis for genetic improvement of wheat resistance,a durum wheat TDU-1 that was susceptible to powdery mildew was used to hybridize with TDI-1,and their F₁ plants,F₂ population,and F_2:3 lines were obtained.Genetic analysis of resistance was conducted on parents TDI-1,TDU-1,and their hybrid offspring that were inoculated with powdery mildew isolate E09.Then,bulked segregant analysis method combined with molecular markers was used to map the resistance gene.The results showed that TDI-1 was susceptible to E09 during the seedling stage but immune during the adult stage.F₁ plants derived from the cross of TDI-1 and TDU-1 were immune to E09 during the adult stage.The resistance of adult F₂individuals was separated,and the ratio of resistant and susceptible plants was 3:1($χ_{3:1}^{2}$=0.11,P=0.74);the ratio of the number of homozygous resistant,separated resistant,and homozygous susceptible F_2:3 lines was 1:2:1($χ_{1:2:1}^{2}$=0.47,P=0.79),indicating that the resistance to powdery mildew in the adult stage of TDI-1 was controlled by one dominant gene,temporarily named PmTDI-1.Subsequently,a set of molecular markers was used to amplify the parents and their F₂ population,and then four markers on chromosome 2A,including Xwmc407,NRM-2AS29,NRM-2AS45 and NRM-2AS84, confirmed to be linked to PmTDI-1. PmTDI-1 was between the flanking markers NRM-2AS45 and NRM-2AS84,with genetics distances of 1.8 cM and 4.6 cM,respectively.Therefore,the adult stage powdery mildew resistance gene PmTDI-1 was preliminarily localized on chromosome 2A.This study identified a novel dominant adult-plant-resistance powdery mildew gene PmTDI-1 from tetraploid wheat TDI-1.

Key words: Tetraploid wheat, Powdery mildew, Resistance gene, Bulked segregant analysis(BSA), Gene mapping

YAN Guiyun, GU Chunxia, WANG Min, TAN Dan, LIU Xiaoyu, LU Chengda, ZUO Jingjing. Molecular Mapping of Adult Plant Resistance Gene to Powdery Mildew in Tetraploid Wheat[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 187-193. doi: 10.7668/hbnxb.20195107.

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Figures/Tables 8

Tab.1 Molecular markers linked to PmTDI-1

引物名称 Primer name	染色体位置/Mb Chromosome location	正向引物(5'—3') Forward sequence (5'—3')	反向引物(5'—3') Reverse sequence(5'—3')	退火温度/℃ Tm
Xwmc407	2A/7.81	GGTAATTCTAGGCTGACATATGCTC	CATATTTCCAAATCCCCAACTC	61
NRM-2AS29	2A/16.90	GGTCTGCGAGCGTTTACT	GCTTTCTTCCTTTCTGCC	58
NRM-2AS45	2A/10.60	GCTAATAACCCGCACTAAA	GCAAACTACCTTACAGAGCA	58
NRM-2AS84	2A/15.36	TGTTGGAGAGAAGGAAAGG	ACGGGAGATGAGGTGTAGT	58

Fig.1 The powdery mildew infection types of TDI-1 and TDU-1 A.Infection types of Bgt isolate E09 on seedlings of TDI-1 and TDU-1;B,C.Infection type of Bgt isolate E09 on TDI-1 and TDU-1 adult plant,respectively.

Tab.2 Grading analysis of TDI-1 and TDU-1 plants to Bgt isolate E09 at seedling stage

亲本 Parents	株数 Number of plants				感病株数Susceptible plants
亲本 Parents	株数 Number of plants	0;	1	2	3	4
TDI-1	10	0	0	0	0	10
TDU-1	10	0	0	0	0	10

Fig.2 Resistance reaction to powdery mildew of adult plants of TDI-1 × TDU-1 F₂ population

Fig.3 Resistance reaction to powdery mildew of adult plants of TDI-1×TDU-1 F_2:3 lines

Tab.3 Chi square analysis of disease resistance in TDI-1×TDU-1 genetic populations

亲本/群体 Parents/ population	株/系数 Number of plants/lines	抗病株/系 Resistant plants/lines	杂合抗病株/系 Heterozygous plants/lines	感病株/系 Susceptible plants/lines	理论比值 Theoretical ratio	χ²	P
TDI-1	10	10	0	0
TDU-1	10	0	0	10
F₂	187	138	0	49	3:1	0.11	0.74
F_2:3	187	51	91	45	1:2:1	0.47	0.79

Fig.4 Genetic map and marker electrophoretic bands of PmTDI-1 M.Ladder Marker;PR.Resistant parent TDI-1;PS.Susceptible parent TDU-1;BR.Resistant bulk;BS.Susceptible bulk;R1—R5.Homozygous resistant individuals;H1—H5.Heterozygous resistant individuals;S1—S5.Homozygous susceptible individuals.

Fig.5 High confidence annotated genes in the candidate region of PmTDI-1

References 37

[1]	何中虎, 庄巧生, 程顺和, 于振文, 赵振东, 刘旭. 中国小麦产业发展与科技进步[J]. 农学学报, 2018, 8(1):107—114.doi:10.11923/j.issn.2095-4050.cjas2018-1-107.
	He Z H, Zhuang Q S, Cheng S H, Yu Z W, Zhao Z D, Liu X. Wheat production and technology improvement in China[J]. Journal of Agriculture, 2018, 8(1):107—114. doi: 10.11923/j.issn.2095-4050.cjas2018-1-107
[2]	Griffey C A. Effectiveness of adult-plant resistance in reducing grain yield loss to powdery mildew in winter wheat[J]. Plant Disease, 1993, 77(6):618.doi:10.1094/pd-77-0618.
[3]	Li Y H, Wei Z Z, Sela H N, Govta L, Klymiuk V, Roychowdhury R, Chawla H S, Ens J, Wiebe K, Bocharova V, Ben-David R, Pawar P B, Zhang Y Q, Jaiwar S, Molnár I, Doležel J, Coaker G, Pozniak C J, Fahima T. Dissection of a rapidly evolving wheat resistance gene cluster by long-read genome sequencing accelerated the cloning of Pm69[J]. Plant Communications, 2024, 5(1):100646.doi:10.1016/j.xplc.2023.100646.
[4]	Huang X Q, Hsam S L K, Zeller F J. Identification of powdery mildew resistance genes in common wheat(Triticum aestivum L.):IX.cultivars,land races and breeding lines grown in China[J]. Plant Breeding, 1997, 116(3):233—238.doi:10.1111/j.1439-0523.1997.tb00988.x.
[5]	He H G, Zhu S Y, Jiang Z N, Ji Y Y, Wang F, Zhao R H, Bie T D. Comparative mapping of powdery mildew resistance gene Pm21 and functional characterization of resistance-related genes in wheat[J]. Theoretical and Applied Genetics, 2016, 129(4):819—829.doi:10.1007/s00122-016-2668-4.
[6]	Matsuoka Y. Evolution of polyploid triticum wheats under cultivation:the role of domestication,natural hybridization and allopolyploid speciation in their diversification[J]. Plant & Cell Physiology, 2011, 52(5):750—764.doi:10.1093/pcp/pcr018.
[7]	He H G, Liu R K, Ma P T, Du H N, Zhang H H, Wu Q H, Yang L J, Gong S J, Liu T L, Huo N X, Gu Y Q, Zhu S Y. Characterization of Pm68,a new powdery mildew resistance gene on chromosome 2BS of Greek durum wheat TRI 1796[J]. Theoretical and Applied Genetics, 2021, 134(1):53—62.doi:10.1007/s00122-020-03681-2.
[8]	Reader S M, Miller T E. The introduction into bread wheat of a major gene for resistance to powdery mildew from wild emmer wheat[J]. Euphytica, 1991, 53(1):57—60.doi:10.1007/BF00032033.
[9]	Rong J K, Millet E, Manisterski J, Feldman M. A new powdery mildew resistance gene:introgression from wild emmer into common wheat and RFLP-based mapping[J]. Euphytica, 2000, 115(2):121—126.doi:10.1023/A:1003950431049.
[10]	Liu Z Y, Sun Q X, Ni Z F, Nevo E, Yang T. Molecular characterization of a novel powdery mildew resistance gene Pm30 in wheat originating from wild emmer[J]. Euphytica, 2002, 123(1):21—29.doi:10.1023/A:1014471113511.
[11]	Blanco A, Gadaleta A, Cenci A, Carluccio A V, Abdelbacki A M M, Simeone R. Molecular mapping of the novel powdery mildew resistance gene Pm36 introgressed from Triticum turgidum var.dicoccoides in durum wheat[J]. Theoretical and Applied Genetics, 2008, 117(1):135—142.doi:10.1007/s00122-008-0760-0.
[12]	Li G Q, Fang T L, Zhang H T, Xie C J, Li H J, Yang T, Nevo E, Fahima T, Sun Q X, Liu Z Y. Molecular identification of a new powdery mildew resistance gene Pm41 on chromosome 3BL derived from wild emmer(Triticum turgidum var.dicoccoides)[J]. Theoretical and Applied Genetics, 2009, 119(3):531—539.doi:10.1007/s00122-009-1061-y.
[13]	Hua W, Liu Z J, Zhu J, Xie C J, Yang T, Zhou Y L, Duan X Y, Sun Q X, Liu Z Y. Identification and genetic mapping of Pm42,a new recessive wheat powdery mildew resistance gene derived from wild emmer(Triticum turgidum var.dicoccoides)[J]. Theoretical and Applied Genetics, 2009, 119(2):223—230.doi:10.1007/s00122-009-1031-4.
[14]	Zhang D Y, Zhu K Y, Dong L L, Liang Y, Li G Q, Fang T L, Guo G H, Wu Q H, Xie J Z, Chen Y X, Lu P, Li M M, Zhang H Z, Wang Z Z, Zhang Y, Sun Q X, Liu Z Y. Wheat powdery mildew resistance gene Pm64 derived from wild emmer(Triticum turgidum var.dicoccoides)is tightly linked in repulsion with stripe rust resistance gene Yr5[J]. The Crop Journal, 2019, 7(6):761—770.doi:10.1016/j.cj.2019.03.003.
[15]	乔麟轶. 小麦材料CH7034中抗白粉病和抗条锈病QTL定位[D]. 太原:山西大学, 2018.doi:10.7666/d.D01594906.
	Qiao L Y. Mapping of resistance loci to powdery mildew and stripe rust in wheat cultivar CH7034[D]. Taiyuan:Shanxi University, 2018.
[16]	盛宝钦, 段霞谕. 对记载小麦成株白粉病 “0-9级法” 的改进[J]. 北京农业科学, 1991(1):38—39.
	Sheng B Q, Duan X Y. Improvement of "0-9 grade method" for recording wheat powdery mildew[J]. Beijing Agricultural Sciences, 1991(1):38—39.
[17]	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.doi:10.1007/s00122-004-1740-7. pmid: 15490101
[18]	Qiao L Y, Zhang X J, Li X, Zhang L, Zheng J, Chang Z J. Development of NBS-related microsatellite(NRM)markers in hexaploid wheat[J]. Euphytica, 2017, 213(11):256.doi:10.1007/s10681-017-2039-5.
[19]	乔麟轶, 常建忠, 郭慧娟, 高建刚, 郑军, 畅志坚. 小麦全基因组NBS类R基因分析及2AL染色体NBS-SSR特异标记开发[J]. 作物学报, 2016, 42(6):795—802.doi:10.3724/SP.J.1006.2016.00795.
	Qiao L Y, Chang J Z, Guo H J, Gao J G, Zheng J, Chang Z J. Genome-wide analysis of TaNBS resistance genes and development of chromosome 2AL-specific NBS-SSR markers in wheat[J]. Acta Agronomica Sinica, 2016, 42(6):795—802.
[20]	Wang B, Meng T, Xiao B, Yu T Y, Yue T Y, Jin Y L, Ma P T. Fighting wheat powdery mildew:from genes to fields[J]. Theoretical and Applied Genetics, 2023, 136(9):196.doi:10.1007/s00122-023-04445-4. pmid: 37606731
[21]	李晓华, 郭慧娟, 畅志坚, 张晓军, 李欣, 乔麟轶, 高伟, 詹海仙. 小麦白粉病成株抗性研究现状[J]. 山西农业科学, 2017, 45(4):653—658,673.doi:10.3969/j.issn.1002-2481.2017.04.40.
	Li X H, Guo H J, Chang Z J, Zhang X J, Li X, Qiao L Y, Gao W, Zhan H X. Research status on adult-plant resistance to wheat powdery mildew[J]. Journal of Shanxi Agricultural Sciences, 2017, 45(4):653—658,673.
[22]	Wang Z L, Li L H, He Z H, Duan X Y, Zhou Y L, Chen X M, Lillemo M, Singh R P, Wang H, Xia X C. Seedling and adult plant resistance to powdery mildew in Chinese bread wheat cultivars and lines[J]. Plant Disease, 2005, 89(5):457—463.doi:10.1094/PD-89-0457. pmid: 30795421
[23]	刘志勇, 张怀志, 白斌, 等. 中国小麦抗条锈病基因育种利用现状与策略[J]. 中国农业科学, 2024, 57(1):34—51.doi:10.3864/j.issn.0578-1752.2024.01.004.
	Liu Z Y, Zhang H Z, Bai B, et al. Current status and strategies for utilization of stripe rust resistance genes in wheat breeding program of China[J]. Scientia Agricultura Sinica, 2024, 57(1):34—51. doi: 10.3864/j.issn.0578-1752.2024.01.004
[24]	乔麟轶, 李锐, 郝宇琼, 乔玲, 李欣, 张晓军, 畅志坚, 郑兴卫. 基因枪介导的冬小麦临汾5064遗传转化体系构建[J]. 山西农业科学, 2023, 51(12):1347—1352.doi:10.3969/j.issn.1002-2481.2023.12.01.
	Qiao L Y, Li R, Hao Y Q, Qiao L, Li X, Zhang X J, Chang Z J, Zheng X W. Construction of a genetic transformation system mediated by biolistic particle for winter wheat Linfen 5064[J]. Journal of Shanxi Agricultural Sciences, 2023, 51(12):1347—1352.
[25]	Sánchez-Martín J, Widrig V, Herren G, Wicker T, Zbinden H, Gronnier J, Spörri L, Praz C R, Heuberger M, Kolodziej M C, Isaksson J, Steuernagel B, Karafiátová M, Doležel J, Zipfel C, Keller B. Wheat Pm4 resistance to powdery mildew is controlled by alternative splice variants encoding chimeric proteins[J]. Nature Plants, 2021, 7(3):327—341.doi:10.1038/s41477-021-00869-2. pmid: 33707738
[26]	Mohler V, Bauer C, Schweizer G, Kempf H, Hartl L. Pm50:a new powdery mildew resistance gene in common wheat derived from cultivated emmer[J]. Journal of Applied Genetics, 2013, 54(3):259—263.doi:10.1007/s13353-013-0158-9.
[27]	Zhu Z D, Kong X Y, Zhou R, Jia J Z. Identification and microsatellite markers of a resistance gene to powdery mildew in common wheat introgressed from Triticum durum[J]. Acta Botanica Sinica, 2004, 46(7):867—872.doi:10.1016/j.molcel.2004.06.032.
[28]	Zhu Z D, Zhou R H, Kong X Y, Dong Y C, Jia J Z. Microsatellite markers linked to 2 powdery mildew resistance genes introgressed from Triticum carthlicum accession PS5 into common wheat[J]. Genome, 2005, 48(4):585—590.doi:10.1139/g05-016.
[29]	Yao D Y, Ijaz W, Liu Y, Hu J H, Peng W T, Zhang B W, Wen X L, Wang J, Qiu D, Li H J, Xiao S H, Sun G Z. Identification of a Pm4 allele as a powdery mildew resistance gene in wheat line Xiaomaomai[J]. International Journal of Molecular Sciences, 2022, 23(3):1194.doi:10.3390/ijms23031194.
[30]	Li G Q, Cowger C, Wang X W, Carver B F, Xu X Y. Characterization of Pm65,a new powdery mildew resistance gene on chromosome 2AL of a facultative wheat cultivar[J]. Theoretical and Applied Genetics, 2019, 132(9):2625—2632.doi:10.1007/s00122-019-03377-2.
[31]	Niu J S, Jia H Y, Yin J, Wang B Q, Ma Z Q, Shen T M. Development of an STS marker linked to powdery mildew resistance genes PmLK906 and Pm4a by gene chip hybridization[J]. Agricultural Sciences in China, 2010, 9(3): 331—336.doi:10.1016/s1671-2927(09)60101-2.
[32]	胡铁柱, 李洪杰, 刘子记, 解超杰, 周益林, 段霞瑜, 贾旭, 尤明山, 杨作民, 孙其信, 刘志勇. 普通小麦品种豫麦66抗白粉病基因的鉴定与分子标记[J]. 作物学报, 2008, 34(4):545—550.doi:10.3321/j.issn:0496-3490.2008.04.002.
	Hu T Z, Li H J, Liu Z J, Xie C J, Zhou Y L, Duan X Y, Jia X, You M S, Yang Z M, Sun Q X, Liu Z Y. Identification and molecular mapping of the powdery mildew resistance gene in wheat cultivar Yumai 66[J]. Acta Agronomica Sinica, 2008, 34(4):545—550.
[33]	Yi Y, Li R F, Xu H X, Wu X Q, Li S P, Zhang J, Yin Y. Identification of SRAP and RGA markers linked to powdery mildew(Blumeria graminis)resistance gene PmZB90 in common wheat[J]. Australian Journal of Crop Science, 2013, 7(3):454—459.
[34]	Xu W G, Li C X, Hu L, Wang H W, Dong H B, Zhang J Z, Zan X C. Identification and molecular mapping of PmHNK54:a novel powdery mildew resistance gene in common wheat[J]. Plant Breeding, 2011, 130(6):603—607.doi:10.1111/j.1439-0523.2011.01882.x.
[35]	Fu B S, Chen Y, Li N, Ma H Q, Kong Z X, Zhang L X, Jia H Y, Ma Z Q. PmX:a recessive powdery mildew resistance gene at the Pm4 locus identified in wheat Landrace Xiaohongpi[J]. Theoretical and Applied Genetics, 2013, 126(4):913—921.doi:10.1007/s00122-012-2025-1.
[36]	Olivera P D, Bulbula W D, Badebo A, Bockelman H E, Edae E A, Jin Y. Field resistance to wheat stem rust in durum wheat accessions deposited at the USDA national small grains collection[J]. Crop Science, 2021, 61(4):2565—2578.doi:10.1002/csc2.20466. pmid: 34413535
[37]	乔麟轶, 畅志坚, 张晓军, 刘静, 朱艳, 詹海仙, 郭慧娟, 李欣. 乌拉尔图小麦全基因组NBS-SSR位点分析及标记开发[J]. 山西农业大学学报(自然科学版), 2017, 37(3):153—157.doi:10.13842/j.cnki.issn1671-8151.2017.03.001.
	Qiao L Y, Chang Z J, Zhang X J, Liu J, Zhu Y, Zhan H X, Guo H J, Li X. Genome-wide analysis and marker development of NBS-SSR in Triticum urartu[J]. Journal of Shanxi Agricultural University(Natural Science Edition), 2017, 37(3):153—157.

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