Distribution and Evaluation of 6 Blast Resistance Genes in Major Cultivated Rice Varieties in Zhejiang
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摘要:
目的 检测浙江省40份水稻栽培品种和6份稻瘟病菌生理小种鉴别品种中6个抗性基因分布情况，同时鉴定品种苗瘟抗性水平，探讨携带抗性基因和抗性水平相关性。 方法 利用功能性分子标记检测品种中Pi1、Pi9/Piz、Pi2/Pizt、Pikh、Pikm、Pita，6个稻瘟病抗性基因的分布情况。用2015-2017年田间采集到的141个稻瘟菌菌株鉴定46个水稻品种苗瘟抗性水平。 结果 6个抗性基因在浙江省栽培品种中分布频率不同，其中Pita基因分布最广，占47.83%；其次是Pikh，占41.30%；抗性基因Pi2/Pizt和Pikm在栽培品种中分布频率相等，均为34.78%；抗性基因Pi9/Piz在栽培品种中分布较少，仅有21.74%；抗性基因Pi1仅在稻瘟病菌生理小种鉴别品种TTP中检测到。抗性水平鉴定结果显示22个品种的抗性频率在60%以上，两个品种的抗性频率达到80%以上。 结论 浙江40份水稻栽培品种和6份稻瘟病菌生理小种鉴别品种携带抗性基因和其抗性水平部分相关。 Abstract:Objective The distribution of 6 resistance genes in 40 cultivated rice varieties in Zhejiang (CRZ) and 6 physiological race identification (PRI) cultivars of Magnaporthe grisea as well as the blast resistance of the rice seedlings were studied. Method The distribution of 6 resistance genes, i.e., Pi1, Pi9/Piz, Pi2/Pizt, Pikh, Pikm and Pita, in the rice varieties was obtained using functional markers. The resistance to M. grisea of all 46 rice varieties was determined based on the 141 strains collected in the field from 2015 to 2017. Result Among the resistance genes, Pita had the highest distribution frequency at 47.83% followed by Pikh at 41.30%; Pi2/Pizt and Pikm, same at 34.78%; Pi9/Piz, low at 21.74%; and Pi1 found only in TTP, one of PRI cultivars. In all, there were 22 rice varieties that showed a resistance rate beyond 60%, and 2 varieties more than 80%. Conclusion The resistance genes carried by the 40 CRZ and the 6 PRI cultivars partially correlated with the resistance level exhibited by the cultivars. -
Key words:
- rice /
- rice leaf blast /
- molecular marker /
- resistance gene
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图 1 6个稻瘟病基因抗感性分子标记检测
注：M为DL2000，A为Pi1基因抗性分子标记检测，CK为IRBL1-CL; B为Pi9/Piz基因抗感分子标记检测，CK为IRBL9-W; C为Pi2/Pizt基因抗性分子标记检测，CK为IRBLz5-CA; D为Pikh基因抗感分子标记检测，CK为IRBLkh-K3; E为Pikm基因抗感分子标记检测，CK为IRBLkm-Ts; F1为 Pita基因抗性分子标记检测，F2为 Pita基因感性分子标记检测，CK为IRBLta2-Pi.第二泳道为阴性对照丽江新团黑谷.泳道1~12为部分浙江省主栽品种材料，依次是Ⅱ优650、Y两优2号、宁81、Y两优1号、Y两优689、春优84、丰两优香1号、嘉58、EX119、浙粳86、内五优8015、钱优0508。
Figure 1. Detection of 6 rice blast resistance genes by specific molecular markers
Note:M: DL2000;A: Identification of Pi1 with resistance molecular markers, CK: IRBL1-CL; B: Identification of Pi9/Piz with resistance and susceptible molecular markers, CK: IRBL9-W; C: Identification of Pi2/Pizt with resistance and susceptible molecular markers, CK: IRBLz5-CA; D: Identification of Pikh with resistance and susceptible molecular markers, CK: IRBLkh-K3; E: Identification of Pikm with resistance molecular markers, CK: IRBLkm-Ts; F1: Identification of Pita with resistance molecular markers; F2: Identification of Pita with susceptibility molecular markers, CK: IRBLta2-Pi. Second line is negative control, LTH. Line 1-12: some main varieties in Zhejiang, i.e., Ⅱ you 650, Y Liangyou 2, Ning 81, Y Liangyou 1, Y Liangyou 689, Chunyou 84, Feng Liangyou 1, Jia 58, EX119, Zhegeng 86, Neiwuyou 8015, Qianyou 0508.
图 2 6个抗稻瘟病基因在浙江主栽水稻品种和稻瘟菌生理小种鉴别品种中的分布
Figure 2. Distribution of 6 rice resistant genes in CRZ and PRI cultivars
表 1 用于PCR扩增的分子标记
Table 1. Functional markers used for resistant gene amplification
基因
Gene引物名称
Primer name引物序列(5′-3′)
Primer sequence(5′-3′)片段大小
Frangment size/bp抗感性
Resistance and susceptibilityPil M-Pi1 F:GAACAATGCCCAAACTTGAGA
R:GGGTCCACATGTCAGTGAGC460 抗病 Pi9/Piz M-Pi9/Piz F:GCTGTGCTCCAAATGAGGAT
R:GCGATCTCACATCCTTTGCT291/397 抗病/感病 Pi2/Pizt M-Pi2/Pizt F:CAGCGATGGTATGAGCACAA
R:CGTTCCTATACTGCCACATCG450/282 抗病/感病 Pikh FM143 F:CCCAACATTGGTAGTAGTGC
R:TCCTTCATACGCAACAATCT258/401 抗病/感病 Pikm Pik-1 F:TCGAGTTGCTGGAACAAGGG
R:ACTGCGTTCCCAATGCATGA575 抗病 Pita Pita-1 F:AGCAGGTTATAAGCTAGGCC
R:CTACCAACAAGTTCATCAAA1042 抗病 Pita-2 F:AGCAGGTTATAAGCTAGGCTAT
R:CTACCAACAAGTTCATCAAA1042 感病 
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表 2 浙江省水稻栽培品种、稻瘟病菌生理小种鉴别品种和丽江新团黑谷单基因系的抗性频率
Table 2. Resistance frequencies of CRZ, PRI and LTH monogenic lines
编号
No.品种名称
Variety
name抗性频率
Resistance
frequency/%1 Ⅱ优650 48.94 2 Y两优2号 59.57 3 宁81 43.26 4 Y两优1号 75.89 5 Y两优689 65.25 6 春优84 76.60 7 丰两优香1号1 30.50 8 嘉58 67.38 9 EX119 21.28 10 浙粳86 79.43 11 内五优8015 65.96 12 钱优0508 67.38 13 钱优1号 48.23 14 钱优930 53.19 15 绍糯9714 58.87 16 深两优5814 71.63 17 秀水12 58.16 18 秀水321 81.56 19 秀优378 63.83 20 浙粳99 79.43 21 甬优12 74.47 22 甬优15 54.61 23 甬优17 59.57 24 甬优538 73.05 25 甬优8 44.68 26 粤优938 38.30 27 浙优12 81.56 28 浙糯优1号 53.90 29 中浙优1号 52.48 30 中浙优8 73.0 31 浙粳70 74.47 32 甲农糯 7.80 33 秀水519 70.21 34 宁84 64.54 35 秀水134 74.47 36 嘉禾218 27.66 37 宁88 79.43 38 甬优1540 78.01 39 甬优9号 56.03 40 春优927 65.96 41 合江18 26.95 42 四丰43 27.66 43 东农363 50.35 44 关东51 42.55 45 TTP 54.61 46 珍龙13 46.10 47 IRBL1-CL(Pi1) 34.75 48 IRBL9-W(Pi9) 60.99 49 IRBLZ5-CA(Pi2) 58.87 50 IRBLkh-k3(Pikh) 41.13 51 IRBLkm-Ts(Pikm) 43.97 52 IRBLta-K1(Pita) 17.73 53 IRBLz-Fu(Piz) 31.91 54 IRBLzt-T(Pizt) 43.26 注:编号1~40为浙江省栽培品种，41~46为稻瘟病菌生理小种鉴别品种，47~54为丽江新团黑谷单基因系。
Note:No. 1-40 are CRZ cultivars; No. 41-46, PRI cultivars; No. 47-54, LTH monogenic lines.
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表 3 Pi1、Pi9/Piz、Pi2/Pizt、Pikh、Pikm和Pita 6个抗性基因在浙江省栽培品种和稻瘟病菌生理小种鉴别品种中的分布及基因型
Table 3. Distribution and genotypes of Pi1, Pi9/Piz, Pi2/Pizt, Pikh, Pikm and Pita in CRZ and PRI cultivars
品种
Variety基因Gene Pi1 Pi9/Piz Pi2/Pizt Pikh Pikm Pita Ⅱ优650 Ⅱ you 650 SS SS SS SS SS SS Y两优2号 Y liangyou 2 SS SS SS SS SS RS 宁81 Ning 81 SS RR SS RR R_ SS Y两优1号 Y liangyou 1 SS SS SS SS SS RS Y两优689 Y liangyou 689 SS SS SS RS SS RS 春优84 Chunyou 84 SS SS RS RS R_ RS 丰两优香1号 Fengyouxiang 1 SS SS SS RS SS SS 嘉58 Jia 58 SS SS RR SS R_ SS EX119 SS SS SS RR SS SS 浙粳86 Zhegeng 86 SS SS RR SS R_ RR 内五优8015 Neiwuyou 8015 SS SS SS RS SS SS 钱优0508 Qianyou 0508 SS SS SS RS R_ SS 钱优1号 Qianyou 1 SS SS SS RS SS SS 钱优930 Qianyou 930 SS SS SS RR R_ SS 绍糯9714 Shaonuo 9714 SS RR SS RR SS RR 深两优5814 Shenyou 5814 SS SS SS SS SS RS 秀水12 Xiushui 12 SS RR SS SS R_ RR 秀水321 Xiushui 321 SS SS RR SS R_ RR 秀优378 Xiuyou 378 SS SS RR SS R_ SS 浙粳99 Zhegeng 99 SS SS RR SS R_ RR 甬优12 Yongyou 12 SS SS RR SS SS RR 甬优15 Yongyou 15 SS SS RS SS R_ SS 甬优17 Yongyou 17 SS SS RR SS R_ SS 甬优538 Yongyou 538 SS SS RR RS SS RR 甬优8 Yongyou 8 SS SS SS SS SS RR 粤优938 Yueyou 938 SS SS SS SS SS RR 浙优12 Zheyou 12 SS SS RR SS SS RR 浙糯优1号 Zhenuoyou 1 SS RR SS RR SS RR 中浙优1号 Zhongzheyou 1 SS SS SS SS SS SS 中浙优8 Zhongzheyou 8 SS SS SS SS SS SS 浙粳70 Zhegeng 70 SS SS RR RR R_ RR 甲农糯 Jianongnuo SS SS SS SS SS SS 宁84 Ning 84 SS SS RR SS R_ RR 秀水519 Xiushui 519 SS RR SS SS R_ RR 秀水134 Xiushui 134 SS RR SS SS R_ SS 嘉禾218 Jiahe 218 SS RR SS SS SS SS 宁88 Ning 88 SS SS RR RS SS RR 甬优1540 Yongyou 1540 SS SS RR RR SS RS 甬优9号 Yongyou 9 SS RS SS RS SS RS 春优927 Chunyou 927 SS SS RS SS SS RS 合江18 Hejiang 18 SS RR SS SS SS SS 四丰43 Sifeng 43 SS SS SS SS SS SS 东农363 Dongnong 363 SS RR SS RR R_ SS 关东51 Guandong 51 SS SS SS SS R_ SS TTP R_ SS * RR SS RR 珍龙13 Zhenglong 13 SS SS SS SS SS SS 注: RR:纯合抗病基因型; SS:纯合感病基因型; RS:杂合基因型；*未得到扩增。
Note:RR: resistant homozygous genotype. SS: susceptible homozygous genotype. RS: heterozygous genotype. *: no amplification.
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