Scientia Agricultura Sinica ›› 2024, Vol. 57 ›› Issue (1): 18-33.doi: 10.3864/j.issn.0578-1752.2024.01.003

• SPECIAL FOCUS: IDENTIFICATION OF NEW WHEAT DISEASES RESISTANCE GENES AND BREEDING APPLICATION • Previous Articles     Next Articles

Identification and Evaluation of Stripe Rust Resistance in 153 Wheat Collections

ZHOU JingWei1(), YE BoWei1(), ZHANG PengFei1, ZHANG YuQing2, HAO Min1, YIN YuRuo1, YUAN Chan1, LI ZhiKang1, LI ShunDa1, XIA XianChun3, HE ZhongHu3, ZHANG HongJun3(), LAN CaiXia1()   

  1. 1 College of Plant Science & Technology, Huazhong Agricultural University/Hubei Hongshan Laboratory, Wuhan 430070
    2 Institute of Food Crops, Hubei Academy of Agricultural Sciences, Wuhan 430072
    3 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Wheat Improvement Center, Beijing 100081
  • Received:2022-07-29 Accepted:2022-09-15 Online:2024-01-01 Published:2024-01-10

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Abstract:

【Objective】Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), significantly reduced wheat production worldwide. Breeding resistant wheat varieties is currently considered to be one of the most economical and effective ways to control this disease. Understanding the resistance level of Chinese and International Maize and Wheat Improvement Center (CIMMYT) wheat breeding materials and the distribution of known disease resistance genes will greatly helpful for discovering the new resistance resources and improving the utilization efficiency of disease resistance genes. 【Method】In the present study, we phenotyped 153 wheat breeding lines derived from China and CIMMYT at both seedling against prevalent Chinese Pst races CYR33 and CYR34. In 2018-2019, 2019-2020 and 2020-2021, using the Pst races CYR33 and CYR34 to identify the materials at the adult plant stages in Ezhou, Hubei. In addition, we used the gene-based or closely linked molecular markers of known stripe rust resistance genes Yr5, Yr9, Yr10, Yr15, Yr17, Yr18, Yr26, Yr29 and YrSP to genotype the whole set of wheat collections. 【Result】We found 10 lines immune against CYR33 at the seedling stage (IT: 0), including seven Chinese cultivars (Shannong 28, Luomai 163, Shimai 13, Zhongyi 6, Tanmai 98-2, Zhongmai 175, Taishan 21) and three CIMMYT lines (CIM-53, CIM-60 and CIM-71). However, only two cultivars, Tanmai 98-1 and Shannong 102, showed immune to CYR34 at the seedling stage. Based on the three years field tests, we found 64 lines showed highly resistance to stripe rust (final disease severity, FDS≤5%), including seven Chinese cultivars and 57 CIMMYT lines. The molecular marker analysis of known stripe rust resistance genes showed that there were 31, 23, 73, 2, 4, 50 and 2 lines carrying resistance genes Yr9, Yr10, Yr17, Yr18, Yr26, Yr29 and YrSP, respectively. None of any lines had Yr5 and Yr15. Based on the phenotype, only CIM-53 showed immune against two races at both seedling and adult plant stages (IT=0, FDS=0) and it might carry the known stripe rust resistance gene combination of Yr17+Yr29 based on the genotype. 【Conclusion】A total of 153 wheat collections from China and CIMMYT were showed adult plant resistance to the prevalent Pst races. Among these, Chinese wheat varieties mainly carry Yr9, Yr10 and Yr26, while CIMMYT wheat line mainly carry Yr17, Yr18 and Yr29, indicating that near-immunity resistance of CIMMYT wheat lines due to combinations of 1-2 moderate seedling resistance gene and 2-3 adult plant resistance genes resulting in durable resistance. Therefore, it is very urgent to expand the resistance sources and identify new resistance genes for pyramiding more genes biotechnology methods to develop new wheat varieties with durable rusts resistance and good agronomic traits. This plays an important role for controlling stripe rust in China by improving the resistance level of wheat variety overall.

Key words: stripe rust, seedling resistance, adult plant resistance, resistance gene, molecular marker, phenotyping of wheat disease

Table 1

Primer sequences of molecular markers for known stripe rust resistance genes and positive checks"

基因
Gene		 阳性对照
Positive checks		 标记名称
Molecular marker		 引物序列
Primer sequence (5′-3′)		 目标片段
Targeted sequence (bp)		 参考文献
Reference
Yr5 Avocet S*6/Yr5 Yr5-R-gene_allele GCGCCCCTTTTCGAAAAAATA 83 [12]
Yr5-alternate_allele CTAGCATCAAACAAGCTAAATA
Yr5-common ATGTCGAAATATTGCATAACATGG
Yr9 Avocet S*6/Yr9 H20F GTTGGAAGGGAGCTCGAGCTG 1598 [13]
H20R GTTGGGCAGAAAGGTCGACATC
Yr10 Avocet S*6/Yr10 SC200-F CTGCAGAGTGACATCATACA 200 [14]
SC200-R TCGAACTAGTAGATGCTGGC
Yr15 Avocet S*6/Yr15 Y15K1_F2 GGAGATAGAGCACATTACAGAC 993 [15]
uhw301R TTTCGCATCCCACCCTACTG
Yr17 Avocet S*6/Yr17 cslVrgal3-F TTATTACCTTGATGAGAAATACAG 383 [16]
cslVrgal3-R CTGAAATTGGGACTAGCGAAATTA
Yr18 Avocet S*6/Yr18 csLv34-F GTTGGTTAAGACTGGTGATGG 150 [17]
csLv34-R TGCTTGCTATTGCTGAATAGT
Yr26 Avocet S*6/Yr26 We173-F GGGACAAGGGGAGTTGAAGC 650 [18]
We173-R GAGAGTTCCAAGCAGAACAC
Yr29 Pavon 76 csLV46-F CGAGACGTCGTCTTCTCTAAC 520 [19]
csLV46-R GTGTATGTGTTGATTCTCCTCG
YrSP Avocet S*6/YrSP YrSP-R-gene_allele GAGAAAATCAGCAGGTGG 129 [12]
YrSP-alternate_allele GAGAAAATCAGCAGGTGC
YrSP-common AGCGAGTTGAGGACATTGGT

Table 2

Phenotyping of stripe rust resistance for 153 wheat varieties (lines) at both seedling and adult plant stages"

抗病类型 Resistant types 苗期抗性CYR33
Seedling resistance
CYR33		 苗期抗性CYR34
Seedling resistance
CYR34		 成株期抗性 19YREZ
Adult plant resistance 19YREZ		 成株期抗性 20YREZ
Adult plant resistance
20YREZ		 成株期抗性 21YREZ
Adult plant resistance 21YREZ
总数
Total		 国内品种Chinese wheat varieties 国际玉米小麦改良中心品系
CIMMYT wheat lines		 总数
Total		 国内品种Chinese wheat varieties 国际玉米小麦改良中心品系
CIMMYT wheat lines		 总数
Total		 国内品种Chinese wheat varieties 国际玉米小麦改良中心品系
CIMMYT wheat lines		 总数
Total		 国内品种Chinese wheat varieties 国际玉米小麦改良中心品系
CIMMYT wheat lines		 总数
Total		 国内品种
Chinese wheat varieties		 国际玉米小麦改良中心品系
CIMMYT wheat lines
免疫
Immunity		 10 7 3 2 2 0 28 12 16 23 2 21 17 0 17
高抗
High resistance		 31 6 25 40 7 33 54 28 26 26 5 21 41 6 35
中抗
Moderate resistance		 45 14 31 12 4 8 29 18 11 35 11 24 38 28 10
中感
Moderate susceptible		 27 17 10 20 10 10 34 31 3 50 45 5 52 43 9
高感
High susceptible		 36 34 2 68 51 17 8 8 0 18 18 0 4 4 0
缺失
Missing		 4 3 1 11 7 4 0 0 0 1 0 1 1 0 1

Table 3

Correlation coefficient of stripe rust in 153 materials under different field environments for 3 years"

年份 Years 19YREZ 20YREZ
20YREZ 0.79**
21YREZ 0.75** 0.85**

Table 4

Identification of 64 advanced wheat lines with high level of resistance to stripe rust over three years field phenotyping and seedling test (mean of final disease severity (FDSM)≤5%)"

品种（系）
Variety (line)		 苗期Seedling 成株期Adult 抗病基因检测Detection of Yr genes
CYR33 CYR34 19YREZ 20YREZ 21YREZ FDSM Yr5 Yr9 Yr10 Yr15 Yr17 Yr18 Yr26 Yr29 YrSP
偃高1号
Yangao 1		 34 - 0 1R 1R 1 - + - - - - - - -
西昌19
Xichang 19		 34 - 0 0 1R 0 - + - - - - - - -
西科麦6号 Xikemai 6 34 - 0 5R 1R 2 - + - - + - - + -
中麦9号
Zhongmai 9		 34 34 1R 5MS 5MR 4 - - + - - - - - -
13213 34 1 0 0 1R 0 - - - - - - + - -
川育26
Chuanyu 26		 67 56 0 1R 5R 2 - - - - + - - + -
川麦604
Chuanmai 604		 56 1 0 10MS 1R 4 - - - - + - + - -
CIM-1 56 1 1R 0 0 0 - - - - + - - + -
CIM-2 56 7 0 0 1R 0 - - - - + + - + -
CIM-3 34 1 0 5R 1MR 2 - - - - + - - + +
CIM-5 56 67 1R 5MR 5MR 4 - - - - + - - + -
CIM-6 56 56 1R 5MR 1R 2 - - - - + - - + -
CIM-7 56 67 1R 1MS 0 1 - - - - + - - + -
CIM-8 56 1 1R 1R 1MS 1 - - - - + - - NA -
CIM-9 56 1 1R 1R 5R 2 - - - - + - - - -
CIM-10 56 8 1R 5R 1R 2 - - - - + - - + -
CIM-12 56 78 1R 5R 1MR 2 - - - - + - - + -
CIM-13 56 89 1R 1MR 1R 1 - - - - + - - NA -
CIM-14 56 9 1R 10MR 5R 5 - - - - + - - + -
CIM-15 56 8 1R 5MR 5MS 4 - - - - + - - + -
CIM-16 56 78 1R 10RMR 1R 4 - - - - + - - + -
CIM-17 56 1 1R 1R 1R 1 - - - - + - - - -
CIM-20 56 1 1R 1R 0 1 - - - - + - - - -
CIM-22 56 7 1R 5MR 5R 4 - - - - + - - + -
CIM-23 34 12 1R 5R 0 2 - - + - + - - + -
CIM-24 34 2 1R 1R 0 1 - - - - - - - + -
CIM-25 34 2 1R 0 1R 1 - - - - - - - + -
CIM-26 34 - 1R 0 1R 1 - - - - + - - - -
CIM-28 34 78 1R 1R 1R 1 - - - - + - - + -
CIM-29 34 45 1R 1R 0 1 - - + - + - - NA -
CIM-30 34 12 1R 5R 5R 4 - - - - + - - - -
CIM-33 34 56 1R 1R 5R 2 - - - - + - - + -
CIM-34 34 1 1R 1RMR 5R 2 - - - - + - - + -
CIM-35 34 - 1R 0 1R 1 - - - - + - - + -
CIM-36 34 67 1R 0 1MR 1 - + - - + - - + -
CIM-37 34 56 1R 1R 0 1 - - - - + - - + -
CIM-38 34 1 1R 0 1R 1 - - - - + - - + -
CIM-39 34 1 1R 0 0 0 - - - - + - - + -
CIM-40 67 89 0 0 5R 2 - - - - + + - + -
CIM-41 34 67 1RMR 1RMR 5MS 2 - - - - + - - - -
CIM-43 8 2 0 0 10MS 3 - - - - + - - + -
CIM-45 45 2 1MR 1MR 5R 2 - - - - + - - - -
CIM-46 67 6 1R 1R 0 1 - - - - + - - + -
CIM-47 34 78 1R 1R 10MS 4 - - - - + - - - -
CIM-48 56 56 0 0 1R 0 - - - - + - - - -
CIM-49 45 1 1MR 1MR 1R 1 - - - - + - - - -
CIM-50 34 1 0 0 0 0 - - - - + - - + -
CIM-51 34 1 0 0 0 0 - - - - + - - - -
CIM-52 56 1 5MS 5MS 1R 4 - - - - + - - + -
CIM-53 0 1 0 0 0 0 - - - - + - - + -
CIM-54 45 78 0 0 0 0 - - - - + - - + -
CIM-55 45 1 5MS 5MS 1R 4 - - - - + - - + -
CIM-56 6 6 0 0 5R 2 - - - - + - - - -
CIM-57 45 56 0 0 1R 0 - - - - + - - + -
CIM-58 67 12 1MR 1MR 1R 1 - - - - + - - + -
CIM-59 NA 34 0 0 0 0 - - - - + - - + -
CIM-60 0 7 0 0 1R 0 - - - - + - - - -
CIM-61 67 8 1MR 1MR 1R 1 - - - - + - - - -
CIM-63 67 1 1MR 1MR 1R 1 - - - - + - - - -
CIM-64 45 67 0 0 0 0 - - - - + - - + -
CIM-65 56 2 5R 5R 5R 5 - - - - + - - NA -
CIM-68 23 1 1R 1R 1R 1 - - - - + - - + -
CIM-70 45 89 1R 1R 0 1 - - - - + - - - -
CIM-72 78 89 0 0 1R 0 - - - - + - - + -

Fig. 1

Detection results of stripe rust resistance gene molecular markers in 153 varieties (lines) M: DNA ladder marker; +: The positive controls are Avocet S*6/Yr9, Avocet S*6/Yr10, Avocet S*6/Yr15, Avocet S*6/Yr17, Avocet S*6/Yr18, Avocet S*6/Yr26 and Pavon76; -: The negative control is Avocet-YRA; a: Amplification of wheat collection using the molecular marker H20 linked to the stripe rust resistance gene Yr9; b: Amplification of wheat collection using the molecular marker SC200 linked to Yr10; c: Amplification of wheat collection using the functional molecular marker Y15K1_F2 of Yr15; d: Amplification of wheat collection using the closely linked molecular marker csVrga13 of Yr17; e: Amplification of wheat collection using the molecular marker csLV34 closely linked to Yr18; f: Amplification of wheat collection using the molecular marker WE173 closely linked to Yr26; g: Amplification of wheat collection by using the molecular marker csLV46 closely linked to Yr29; No. 1-22, 45-66, 67-88, 89-97 same as Supplementary table 1"

Fig. 2

Amplification of wheat collection using the functional molecular marker Yr5-R-gene_allele (a) and YrSP R-gene_allele (b) for the stripe rust resistance genes Yr5 and YrSP a: Black dot: ddH2O; orange: Positive controls Avocet S*6/Yr5; blue: Negative control Avocet-YrA; green: Test material. b: Black: ddH2O; orange: Negative control Avocet-YrA; blue: Positive controls Avocet S*6/YrSP; green: Test material"

Fig. 3

The gene combinations of known stripe rust resistance gene in 153 wheat collection"

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