Scientia Agricultura Sinica ›› 2024, Vol. 57 ›› Issue (13): 2568-2582.doi: 10.3864/j.issn.0578-1752.2024.13.006

• PLANT PROTECTION • Previous Articles     Next Articles

Stem Rust Resistance Genes Identification and Evaluation of 88 Wheat Cultivars (Lines) in Heilongjiang Province

GAO Fu1(), WANG Rui1(), LIU DongJun2, SUN HuiYan1, WANG ZiYe1, SONG WeiFu2, LI TianYa1()   

  1. 1 College of Plant Protection, Shenyang Agricultural University, Shenyang 110866
    2 Institute of Crop Resources, Heilongjiang Academy of Agricultural Sciences, Harbin 150086
  • Received:2024-01-21 Accepted:2024-02-16 Online:2024-07-09 Published:2024-07-09
  • Contact: LI TianYa

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Abstract:

【Objective】 In recent years, the new races of Puccinia graminis f. sp. tritici (Pgt), Ug99, TKTTF, and TTRTF, have emerged and continuously spread. Heilongjiang Province is an area where wheat stem rust occurred frequently. The aim of this study is to analyze the resistance genes contained in wheat cultivars in this region and their resistance to predominant races of Pgt, and to provide a theoretical basis for the rational regional layout of resistance genes and the breeding of disease resistant cultivars in China. 【Method】 Based on the response types of 43 monogenic lines and 88 wheat cultivars (lines) to 7 different virulence races (21C3CTTSC, 21C3CTHQM, 34MKQSM, 34C3MTGQM, 34MKGQM, 34MTGSM, and RTGRM) of Pgt, the stem rust resistance genes contained in these 88 cultivars (lines) were postulated; Further, the presence of Sr31, Sr24, Sr25, Sr26, and Sr38 in the tested cultivars (lines) was detected by using molecular markers closely linked with these five genes. At the same time, the resistance of these cultivars (lines) to two predominant races 21C3CTHQM and 34MKGQM was evaluated at the adult-plant stage during the period of 2021-2022.【Result】 The results of gene postulation showed that 37 wheat cultivars might contain one or more of the wheat stem rust resistance genes Sr9e, Sr11, Sr17, Sr23, Sr30, Sr35, Sr37, Sr39, Sr40, and SrWld. In addition, molecular testing confirmed that 10 wheat cultivars (Kenmai 16, Longchun 204, Longfumai 8171, Longmengmai 2305, Longfumai 7166, Longchun 218, Longmai 95, Longfumai 19-92, Kechun 171286, and Longfumai 194) contained the resistance gene Sr31, but none of wheat cultivars (lines) carried Sr24, Sr25, Sr26, and Sr38. In field trials in 2021 and 2022, 86 (97.7%) and 83 (94.3%) wheat cultivars showed resistance to race 21C3CTHQM and 34MKGQM, respectively. Among them, 62 (70.5%) wheat cultivars showed good resistance to both tested races for two consecutive years, while the other cultivars (lines) showed different degrees of susceptibility to a single race.【Conclusion】 Wheat cultivars (lines) in Heilongjiang Province have good resistance to the tested races at adult-plant stage, and 37 wheat cultivars may carry one or more of the 10 resistance genes (Sr9e, Sr11, Sr17, Sr23, Sr30, Sr35, Sr37, Sr39, Sr40, and SrWld). There are 10 cultivars (lines) containing Sr31, and no wheat cultivars (lines) containing Sr24, Sr25, Sr26 and Sr38 resistance genes were detected.

Key words: wheat stem rust, Puccinia graminis f. sp. tritici (Pgt), molecular marker, resistance gene, cultivar

Table 1

PCR primers used for molecular markers"

抗性基因
Sr gene		 引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 参考文献
Reference
Sr24 Sr24#50 CCCAGCATCGGTGAAAGAA
ATGCGGAGCCTTCACATTTT		 [18]
Sr25 Xwmc221 ACGATAATGCAGCGGGGAAT
GCTGGGATCAAGGGATCAAT		 [19]
Sr26 Sr26#43 AATCGTCCACATTGGCTTCT
CGCAACAAAATCATGCACTA		 [18]
Sr31 SCSS30.2576 GTCCGACAATACGAACGATT
CCGACAATACGAACGCCTTG		 [20]
Iag95 CTCTGTGGATAGTTACTTGATCGA
CCTAGAACATGCATGGCTGTTACA		 [21]
Sr38 VENTRIUP-LN2 AGGGGCTACTGACCAAGGCT
TGCAGCTACAGCAGTATGTACACAAAA		 [22]

Table 2

PCR amplification conditions for Sr genes molecular marker"

抗性基因
Sr gene		 引物名称
Primer name		 PCR扩增条件PCR amplification condition
温度Temperature (℃)/时间Time 循环数Number of cycles
Sr24 Sr24#50 94/3 min 1
94/30 s; 57/30 s; 72/40 s 30
20/1 min 1
Sr25 Xwmc221 94/3 min 1
94/30 s; 60/30 s; 72/40 s 30
20/1 min 1
Sr26 Sr26#43 94/3 min 1
94/30 s; 56/30 s; 72/40 s 30
20/1 min 1
Sr31
 SCSS30.2576 95/5 min 1
95/1 min; 60/1 min; 72/30 s 35
72/10 min 1
Iag95 94/3 min 1
94/30 s; 55/1 min; 72/70 s 30
25/1 min 1
Sr38 VENTRIUP-LN2 94/45 s 1
94/45 s; 65/30 s; 72/1 min 30
72/7 min 1

Table 3

Resistance levels of 88 wheat cultivars (lines) to races 21C3CTHQM, and 34MKGQM of Pgt at adult-plant stage"

品种（系）
Cultivar (line)		 抗性水平Resistance level
34MKGQM 21C3CTHQM
2021 2022 2021 2022
龙麦51 Longmai 51 5R5 5R5 10R10 10R10
龙麦61 Longmai 61 0 0 0 0
龙麦62 Longmai 62 100MR100 100MR100 0 0
龙麦63 Longmai 63 0 0 0 0
龙麦64 Longmai 64 0 0 0 0
龙麦65 Longmai 65 0 0 0 0
龙麦60 Longmai 60 0 0 0 5R5
龙麦67 Longmai 67 0 0 0 0
龙麦68 Longmai 68 0 0 0 0
龙麦69 Longmai 69 50R10 50R10 0 0
龙麦70 Longmai 70 0 0 0 0
克旱5 Kehan 5 50MR30 50MR50 80R80 80R80
克旱7 Kehan 7 0 0 80R80 80R80
垦红14 Kenhong 14 0 5R5 80R80 80R80
龙麦19 Longmai 19 50R50 50R50 0 5R5
龙麦28 Longmai 28 0 0 0 0
龙麦77 Longmai 77 0 0 0 0
龙麦94 Longmai 94 70MS100 80MS80 5MR10 50R50
龙麦7574 Longmai 7547 0 0 0 0
龙麦89 Longmai 89 0 0 0 0
龙麦7765 Longmai 7765 50MR70 50MR70 0 0
龙麦7202 Longmai 7202 0 0 0 0
龙麦87 Longmai 87 0 0 0 0
龙麦92 Longmai 92 50MS30 50MS50 0 0
龙麦96 Longmai 96 0 0 0 0
龙麦97 Longmai 97 0 0 0 0
龙麦93 Longmai 93 0 80NIM80 50R50 50R50
龙麦90 Longmai 90 0 0 0 0
龙麦H336 Longmai H336 0 0 0 0
龙麦85 Longmai 85 0 0 0 0
龙麦81 Longmai 81 0 0 0 0
龙麦7117 Longmai 7117 30MR70 30MR70 0 0
龙麦86 Longmai 86 0 0 0 0
龙麦7507 Longmai 7507 0 0 0 0
龙麦91 Longmai 91 0 0 0 0
龙麦95 Longmai 95 0 0 0 0
龙麦76 Longmai 76 0 0 0 0
2021 2022 2021 2022
龙麦15 Longmai 15 0 0 0 0
龙麦29 Longmai 29 70S10 70S10 0 0
垦大3号 Kenda 3 100MS100 100MS80 30S50 80S80
克旱6号 Kehan 6 0 0 80R80 80R80
龙10-0632 Long 10-0632 0 5R5 0 0
龙麦20 Longmai 20 0 0 0 0
龙春2 Longchun 2 0 0 0 0
东农173547 Dongnong 173547 0 0 0 0
克春140891 Kechun 140891 0 0 0 0
克春150397 Kechun 150397 0 0 50R50 50R50
克春1501185 Kechun 1501185 0 0 0 0
克春151350 Kechun 151350 0 0 0 0
克春161727 Kechun 161727 0 0 80R50 80R50
克春1601038 Kechun 1601038 0 0 0 50R50
克春160478 Kechun 160478 0 0 0 0
克春170250 Kechun 170250 0 0 0 0
克春170375 Kechun 170375 0 0 0 5R5
克春171286 Kechun 171286 0 0 0 0
克春171470 Kechun 171470 0 0 0 0
克春18Z762 Kechun 18Z762 0 0 0 0
龙辐麦194 Longfumai 194 0 0 0 0
龙辐麦159 Longfumai 159 0 0 0 50R10
龙辐麦97 Longfumai 97 0 0 0 0
龙辐麦7166 Longfumai 7166 0 0 0 0
龙辐麦8171 Longfumai 8171 0 0 0 0
龙辐麦8396 Longfumai 8396 0 0 50R50 50R50
龙辐麦196 Longfumai 196 0 0 0 0
龙辐17-232 Longfu 17-232 0 0 100S100 100S80
龙辐麦19-92 Longfumai 19-92 0 0 0 0
龙辐19-285 Longfu 19-285 0 0 0 0
龙麦8511 Longmai 8511 0 0 0 0
龙麦8267 Longmai 8267 0 0 0 0
龙蒙麦7255 Longmengmai 7255 0 0 0 0
龙蒙麦8654 Longmengmai 8654 0 0 0 0
龙蒙麦2305 Longmengmai 2305 0 0 0 0
龙麦4062 Longmai 4062 0 0 0 0
龙麦8156 Longmai 8156 0 0 0 0
龙麦8159 Longmai 8159 0 0 0 0
2021 2022 2021 2022
龙麦2018 Longmai 2018 0 0 0 0
垦麦17 Kenmai 17 50MS80 50MS80 0 0
垦麦16 Kenmai 16 0 0 0 0
龙春175 Longchun 175 0 0 0 0
龙春201 Longchun 201 0 0 0 0
龙春203 Longchun 203 0 0 0 0
龙春204 Longchun 204 0 0 0 0
龙春208 Longchun 208 0 0 0 0
龙春211 Longchun 211 0 0 0 0
龙春212 Longchun 212 0 0 0 0
龙春215 Longchun 215 0 0 0 0
龙春216 Longchun 216 0 0 0 0
龙春218 Longchun 218 0 0 0 0

Fig. 1

The amplification results of some wheat cultivars (lines) in Heilongjiang Province with primers Iag95 and SCSS30.2576"

Table 4

Infection type of seven tested races to monogenic line wheat cultivars (Host A)"

单基因系
Monogenic line		 侵染型Infection type
34MTGSM 34C3MTGQM 34MKQSM 21C3CTTSC 21C3CTHQM RTGRM 34MKGQM
Sr5 4 4 4 0 1 4 4
Sr21 2 0 4 1 0 4 1
Sr9e 2 1 1 2 0 1 ;
Sr7b 4 4 4 4 3 4 4
Sr11 3 3 0; 4 3 4 1
Sr6 4 4 4 4 4 4 4
Sr8a 4 4 4 3 4 4 4
Sr9g 4 4 4 4 4 4 4
Sr36 0 2 4 3 2 2 0
Sr9b 4 3 4 4 4 4 4
Sr30 2 2 1 4 0 1 2
Sr17 2 0 1 4 3 0 0
Sr9a 4 4 4 4 4 4 4
Sr9d 4 4 4 4 4 4 4
Sr10 4 3 4 4 0 1 1
Tmp 2 2 0 0 1 4 2
Sr24 4 4 4 0 0 4 4
Sr31 2 2 0 2 1 2 2
Sr38 2 1 1 1 0 1 1
Sr39 4 4 1 4 0 4 0;
Sr40 2 2 2 2 2 2 2
Sr44 2 2 2 2 2 2 4
Sr9f 4 4 4 4 4 4 4
Sr12 4 4 4 4 4 4 4
Sr13 4 4 2 4 4 3 4
Sr15 4 4 2 4 4 4 4
Sr16 4 4 4 4 4 4 4
Sr18 4 4 0 4 4 4 4
Sr19 4 4 2 4 4 4 4
Sr22 4 4 3 4 4 4 4
Sr23 3 4 2 4 1 4 4
Sr25 2 3 4 4 0 1 4
Sr26 4 3 0 4 2 2 4
Sr27 4 4 2 4 0 4 4
Sr28 4 4 3 4 4 4 4
Sr32 4 4 3 4 4 4 4
Sr33 0 2 1 1 0 2 2
Sr34 4 4 3 4 4 4 4
Sr35 4 4 4 3 0 4 4
Sr37 4 2 4 3 1 4 4
Tt-3 0 1 2 2 2 2 2
Wld 3 2 4 2 1 3 4
dp-2 4 4 4 4 4 4 4

Table 5

Infection type of seven tested races to 88 wheat cultivars (lines) (Host B)"

品种（系）
Cultivar (line)		 侵染型Infection type
34 34C3 21C3
MTGSM MKGQM MKQSM MTGQM CTTSC CTHQM RTGRM
龙麦70 Longmai 70 0; 2 1 1 1 0 0;
龙麦7117 Longmai 7117 0; 1 2 1 1 3 1
龙麦90 Longmai 90 0; 2 1 2 2 0 2
龙麦81 Longmai 81 0; 1 1 0; 2 0 0
龙麦51 Longmai 51 0; 1 1 1 1 0 0;
龙麦97 Longmai 97 3 2 1 2 3 0 1
龙麦68 Longmai 68 2 2 1 1 2 0 1
龙麦15 Longmai 15 3 1 2 1 2 0 0;
龙麦65 Longmai 65 0; 1 0 1 1 0 0;
龙麦93 Longmai 93 0; 0 1 0 0 0 1
龙麦76 Longmai 76 2 3 2 1 3 0 2
龙麦H336 Longmai H336 0 1 0; 0 0 0 0
龙麦7547 Longmai 7547 3 2 1 1 1 0 1
龙麦96 Longmai 96 0; 2 0 2 1 0 1
龙麦60 Longmai 60 0; 2 1 1 1 0 0;
龙麦19 Longmai 19 0; 0; 0 0; 0; 0 0
龙麦63 Longmai 63 0; 0; 1 1 1 0 1
龙麦87 Longmai 87 4 4 4 4 4 2 1
龙麦7202 Longmai 7202 0; 1 1 1 2 0 0;
龙麦29 Longmai 29 1 1 1 1 1 2 1
克旱7 Kehan 7 1 3 4 1 3 0 3
龙麦7507 Longmai 7507 1 2 3 1 1 0; 3
垦大3号 Kenda 3 0; 1 2 2 2 0; 3
龙麦86 Longmai 86 1 3 3 3 3 0; 3
龙麦62 Longmai 62 0; 1 1 1 0 0; 2
龙麦28 Longmai 28 0; 1 0 1 0; 0; 0
龙麦85 Longmai 85 0; 0 1 0 1 0 2
龙麦92 Longmai 92 2 4 4 4 1 0 4
龙麦7765 Longmai 7765 2 4 4 4 3 0 3
龙麦64 Longmai 64 2 3 1 1 1 0 0;
龙麦94 Longmai 94 0; 3 0; 1 1 0; 3
龙10-0632 Long 10-0632 2 3 1 1 2 0; 3
龙春2号 Longchun 2 0; 4 2 3 2 0 3
龙麦89 Longmai 89 0; 0; 0; 0; 0 0 0
龙麦20 Longmai 20 0 0 1 0 4 0 0
克旱5 Kehan 5 4 4 4 4 4 0; 4
克春150397 Kechun 150397 2 4 0; 2 1 0; 0;
克春170375 Kechun 170375 0 4 0 3 2 0 2
龙春201 Longchun 201 1 4 0; 0 2 0; 2
克春160478 Kechun 160478 0 1 0 2 1 0 0;
龙麦61 Longmai 61 3 1 4 3 1 0 1
克旱6 Kehan 6 3 1 4 3 1 0 2
龙春215 Longchun 215 4 1 4 4 1 0; 3
龙麦77 Longmai 77 4 2 4 2 3 0; 2
垦麦17 Kenmai 17 2 3 3 2 3 0 2
龙麦95 Longmai 95 1 3 0 2 2 0 2
龙春208 Longchun 208 0 0; 0 4 0 0 3
龙春203 Longchun 203 3 3 3 1 0; 0 0
龙麦2018 Longmai 2018 0 0 0; 0 0 0 0
龙蒙麦8654 Longmengmai 8654 0; 1 3 1 0; 0 0
垦麦16 Kenmai 16 0 0 0 0 0 2 0
龙辐19-92 Longfu 19-92 0 0 0 0 0 4 0
克春171286 Kechun 171286 0 1 0 2 1 0 0
克春171470 Kechun 171470 0 2 0 2 0 0 0
龙辐17-232 Longfu 17-232 0 0; 0 1 0 0 0
龙麦67 Longmai 67 4 4 4 2 3 0 1
克春18Z762 Kechun 18Z762 0 2 0 0; 0 0 0
东农173547 Dongnong 173547 3 3 4 2 2 0 0;
龙春175 Longchun 175 0 0 0 0 0 0 0
龙麦4062 Longmai 4062 3 3 0 3 3 0 3
龙麦8519 Longmai 8519 0 0 0 0 0 0 0
龙春204 Longchun 204 2 2 1 0; 2 1++ 0
龙麦91 Longmai 91 2 1 4 1 3 3 1
龙春216 Longchun 216 3 2 4 2 4 0 0;
龙辐麦194 Longfumai 194 0; 2 0 2 2 2 0;
龙辐麦196 Longfumai 196 4 2 4 2 3 0 1
龙麦8156 Longmai 8156 0 3 0 1 2 0 0
龙辐19-285 Longfu 19-285 0; 0; 0 1 3 0 0;
龙辐麦8396 Longfumai 8396 1 0 1 2 0; 0 0;
龙春211 Longchun 211 2 4 4 1 2 0 0
龙辐麦8171 Longfumai 8171 2 0; 2 1 1 0 1
克春1501185 Kechun 1501185 4 4 3 4 4 4 4
克春161727 Kechun 161727 4 1 4 3 3 1 2
龙蒙麦7255 Longmengmai 7255 0; 0 0; 0 1 0 0
龙蒙麦2305 Longmengmai 2305 1 2 2 2 1+ 0 0;
龙辐麦7166 Longfumai 7166 2 1 0 0; 2 0 0;
龙麦69 Longmai 69 3 2 3 2 4 0 2
龙麦8511 Longmai 8511 3 1 2 1 2 0 1
龙辐麦159 Longfumai 159 3 1 1 1 2 0 1
克春170250 Kechun 170250 0 0 0 0; 0 0 0
龙春212 Longchun 212 0 2 0 2 4 0 2
龙辐麦97 Longfumai 97 0 2 0 1 4 0 3
龙春218 Longchun 218 0 1 1 0 0 0 0
垦红14 Kenhong 14 3 3 3 0 1 3 2
克春140891 Kechun 140891 0 2 1 2 1 0 0
克春1601038 Kechun 1601038 0 0 1 1- 1 0 0
龙麦8267 Longmai 8627 1 0 2 3 1 0 0
克春151350 Kechun 151350 1 0 1 0 1 1 0
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