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作物杂志,2021, 第4期: 59–66 doi: 10.16035/j.issn.1001-7283.2021.04.009

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

黑龙江省粳稻品种稻瘟病主效抗性基因鉴定与抗性评价

高清1(), 张亚玲1, 周弋力1, 于连鹏1, 聂强2, 靳学慧1()   

  1. 1黑龙江八一农垦大学/黑龙江省植物抗性研究中心,163319,黑龙江大庆
    2北大荒农垦集团有限公司建三江分公司农业发展部,156300,黑龙江佳木斯
  • 收稿日期:2020-08-31 修回日期:2020-11-19 出版日期:2021-08-15 发布日期:2021-08-13
  • 通讯作者: 靳学慧
  • 作者简介:高清,主要从事植物病理学研究,E-mail：gqing1198@163.com
  • 基金资助:
    黑龙江省自然科学基金(QC2011C046);黑龙江省农垦总局科技攻关计划(HNK125A-08-06);黑龙江省农垦总局科技攻关计划(HNKXIV-01-04-02);黑龙江省农垦总局科技攻关计划(HKKYZD190205);黑龙江省教育厅项目(12521376);黑龙江八一农垦大学学成、引进人才科研启动计划资助项目(XDB201605);黑龙江八一农垦大学学成、引进人才科研启动计划资助项目(XDB201802);黑龙江八一农垦大学研究生创新科研项目(YJSCX2019-Y18)

Identification of Major Resistance Genes and Resistance Evaluation to Rice Blast in Japonica Rice Varieties in Heilongjiang Province

Gao Qing1(), Zhang Yaling1, Zhou Yili1, Yu Lianpeng1, Nie Qiang2, Jin Xuehui1()   

  1. 1Heilongjiang Bayi Agricultural University/Heilongjiang Plant Resistance Research Center, Daqing 163319, Heilongjiang, China
    2Agricultural Development Department, Jiansanjiang Branch of Beidahuang Agricultural Reclamation Corporation, Jiamusi 156300, Heilongjiang, China
  • Received:2020-08-31 Revised:2020-11-19 Online:2021-08-15 Published:2021-08-13
  • Contact: Jin Xuehui

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摘要：

为明确黑龙江省粳稻品种中稻瘟病抗性基因的类型、评价品种及抗瘟基因利用价值,利用8个已克隆主效抗性基因Pita、Pia、Piz-t、Pib、Pikm、Pi9、Pii和Pid3的特异性分子标记,结合402个黑龙江省各稻区的菌株接种供试品种的抗性表型,对20个黑龙江省粳稻品种的抗性基因型及抗病性进行分析。结果显示,Pia检出率最高,20个粳稻品种均检测到该基因,其次是Pita和Piz-t,检出率为80%;Pia、Pita、Piz-t、Pikm和Pi9在不同生态型品种育种中得到了较为广泛的应用,Pib、Pii和Pid3在不同生态类型品种间分布存在差异,仅在第1积温带的品种中发挥较好作用;唯一携带Pid3且具有Pita+Pia+Piz-t+Pib+Pii+Pid3基因型的龙洋16抗性表现最好,抗性频率高达93%;携带Pita+Pia+Piz-t基因组合的品种均表现出较好的抗病性,对黑龙江省粳稻品种的抗瘟贡献较大。揭示了黑龙江省20个粳稻品种的稻瘟病主效抗性基因类型及其对稻瘟病抗性的贡献,为寒地水稻种质资源的抗病性筛选和广谱抗病基因的利用提供重要依据。

关键词: 稻瘟病, 抗性基因, 抗性评价

Abstract:

In order to clarify the patterns of rice blast resistance genes in the japonica rice varieties in Heilongjiang province and evaluate the utilization value of varieties and the blast resistance genes, this study used specific molecular markers of eight cloned major resistance genes of Pita, Pia, Piz-t, Pib, Pikm, Pi9, Pii, and Pid3, combined with the resistance phenotypes identification of tested varieties inoculated with 402 strains from various rice regions in Heilongjiang, the genotypes and disease resistance of 20 japonica rice varieties in Heilongjiang were analyzed. The results showed that the detection frequency of Pia was the highest, the gene was detected in 20 japonica rice varieties, followed by Pita and Piz-t with detection frequency of 80%; Pia, Pita, Piz-t, Pikm and Pi9 were widely used in the breeding of different ecotype varieties, Pib, Pii, and Pid3 had different distributions among different ecotype varieties, and they only played a good role in varieties suitable for planting in the first accumulated temperature zone; Longyang 16, the only variety carrying Pid3 and having Pita+Pia+Piz-t+Pib+Pii+Pid3 genotype showed the best resistance, and the resistance frequency was 93%. The varieties carrying Pita+Pia+Piz-t gene combinations showed the good disease resistance and made a significant contribution to the blast resistance of japonica rice varieties in Heilongjiang. This study revealed the major blast resistance genetypes of 20 japonica rice varieties in Heilongjiang and their contribution to rice blast resistance, which provided an important basis for disease resistance screening of rice germplasm resources in cold regions and utilization of broad-spectrum resistance genes.

Key words: Rice blast, Resistance gene, Resistance evaluation

图1

参试菌株采集地点示意图

表1

特异性分子标记的引物信息

抗性基因
Resistance gene		 引物
Primer		 引物序列
Primer sequence (5'-3')		 片段大小
Fragment size (bp)		 酶
Enzyme		 参考文献
Reference
Pita YL155-F AGCAGGTTATAAGCTAGGCC 1042(R) [7-9]
YL87-R CTACCAACAAGTTCATCAAA
YL183-F AGCAGGTTATAAGCTAGCTAT 1042(S)
YL87-R CTACCAACAAGTTCATCAAA
Pia Pia-F GTCACGACGACCATCCAGTCAGTAG 2196(R) [10]
Pia-R AGGTAACAGGGAGTGTAGGAAATCA
Piz-t Piz-t-F1 CTTCTATGCAGTCGTGAGATTGGTT 206(R) [10]
Piz-t-R1 ATGCTACGATGGAAATCAACACAAT
Piz-t-F2 TAAAAGATTTCTTGATTTCACTCGG 3154(R)
Piz-t-R2 ACCTGATGGCTAATACTGAACAAGA
Piz-t-F3 GGGCGATGATTTCAACTCACTTTTC 136(R)
Piz-t-R3 GATCATCATGTCCCTCCGAGTCAGA
Pib Pibdom-F GAACAATGCCCAAACTTGAGA 365(R) [11-13]
Pibdom-R GGGTCCACATGTCAGTGAGC
Lys145-F TCGGTGCCTCGGTAGTCAGT 803(S)
Lys145-R GGGAAGCGGATCCTAGGTCT
Pikm Ckm1-F TGAGCTCAAGGCAAGAGTTGAGGA 174 (R)/213 (S) [14-15]
Ckm1-R TGTTCCAGCAACTCGATGAG
Ckm2-F CAGTAGCTGTGTCTCAGAACTATG 290 (R)/332 (S)
Ckm2-R AAGGTACCTCTTTTCGGCCAG
Pi9 Pi9-F GCTGTGCTCCAAATGAGGAT 291 (R)/397(S) [16]
Pi9-R GCGATCTCACATCCTTTGCT
Pii Pii-4SNP-F TCCAATGCTTCTGAAAGGTAGC 353/240+113 PvuⅡ [17]
Pii-4SNP-R TGGAAACATGAACCCATATCC
Pid3 Pid3-F TACTACTCATGGAAGCTAGTTCTC 658 BamHⅠ [18]
Pid3-R ACGTCACAAATCATTCGCTC

表2

供试水稻品种的抗性频率及抗性评价

抗性评价（占比）
Resistance evaluation (percentage)		 品种
Variety		 抗性频率
Resistance
frequency (%)
R (35%) 龙洋16 93.0
龙洋1号 91.0
龙粳40 90.9
北稻4号 89.0
垦稻23 86.3
龙粳42 85.6
北稻3号 81.6
MR (60%) 松粳20 79.6
龙粳43 78.6
绥粳4号 76.6
龙粳31 74.9
龙庆稻3号 74.2
松粳3号 69.9
垦稻12 68.9
稻花香2号 66.2
龙粳21 64.9
龙粳29 63.5
龙粳26 61.5
松粳15 60.9
S (5%) 空育131 36.3

图2

部分水稻品种中稻瘟病抗性基因的扩增产物 M-DL 2000。1-各基因的抗病对照;2-感病对照（LTH）;3-龙粳21;4-龙粳26;5-龙粳29;6-龙粳31;7-龙粳40;8-龙粳42;9-龙粳43;10-垦稻12;11-空育131;12-绥粳4号

表3

供试水稻品种抗性基因分子标记检测结果

编号Number 品种
Variety		 抗性基因Resistance gene 基因型
Genotype
Pita Pia Piz-t Pib Pikm Pi9 Pii Pid3
1 龙洋1号 + + + - + - - - Pita+Pia+Piz-t+Pikm
2 龙洋16 + + + + - - + + Pita+Pia+Piz-t+Pib+Pii+Pid3
3 松粳3号 + + + - - + + - Pita+Pia+Piz-t+Pi9+Pii
4 松粳15 - + + + + + - - Pia+Piz-t+Pib+Pikm+Pi9
5 松粳20 + + + + + + + - Pita+Pia+Piz-t+Pib+Pikm+Pi9+Pii
6 稻花香2号 + + - - - + + - Pita+Pia+Pi9+Pii
7 龙粳21 - + + - + + - - Pia+Piz-t+Pikm+Pi9
8 龙粳42 + + + - - - - - Pita+Pia+Piz-t
9 北稻3号 + + + - + + + - Pita+Pia+Piz-t+Pikm+Pi9+Pii
10 北稻4号 + + + + + + - - Pita+Pia+Piz-t+Pib+Pikm+Pi9
11 绥粳4号 + + - - - + - - Pita+Pia+Pi9
12 垦稻12 + + + - + - - - Pita+Pia+Piz-t+Pikm
13 垦稻23 + + + + - - - - Pita+Pia+Piz-t+Pib
14 龙粳26 - + + - + - - - Pia+Piz-t+Pikm
15 龙粳29 + + - - - + + - Pita+Pia+Pi9+Pii
16 龙粳31 + + + - + - - - Pita+Pia+Piz-t+Pikm
17 龙粳40 + + + - - - - - Pita+Pia+Piz-t
18 龙粳43 + + + + - + + - Pita+Pia+Piz-t+Pib+Pi9+Pii
19 龙庆稻3号 + + + - - + - - Pita+Pia+Piz-t+Pi9
20 空育131 - + - - + + - - Pia+Pikm+Pi9
检出率Detection frequency (%) 80 100 80 30 50 60 35 5

图3

抗性基因在不同生态型粳稻品种中的分布

表4

供试粳稻品种所携带目标基因数及所占比率

目标基因数
Number of
target genes		 基因型数
Number of genotypes		 品种数量
Number of varieties		 比率
Ratio
(%)
3 4 5 25.0
4 5 8 40.0
5 2 2 10.0
6 4 4 20.0
7 1 1 5.0

表5

不同抗性基因类型的稻瘟病抗性表现

基因型
Genotype		 抗性表现
Resistance
reaction		 品种数量
Number of
varieties
Pita+Pia+Piz-t+Pib+Pii+Pid3 R 1
Pita+Pia+Piz-t+Pikm R 1
Pita+Pia+Piz-t R 2
Pita+Pia+Piz-t+Pib+Pikm+Pi9 R 1
Pita+Pia+Piz-t+Pib R 1
Pita+Pia+Piz-t+Pikm+Pi9+Pii R 1
Pita+Pia+Piz-t+Pib+Pikm+Pi9+Pii MR 1
Pita+Pia+Piz-t+Pib+Pi9+Pii MR 1
Pita+Pia+Piz-t+Pikm MR 2
Pita+Pia+Piz-t+Pi9 MR 1
Pita+Pia+Piz-t+Pi9+Pii MR 1
Pita+Pia+Pi9+Pii MR 2
Pita+Pia+Pi9 MR 1
Pia+Piz-t+Pikm+Pi9 MR 1
Pia+Piz-t+Pikm MR 1
Pia+Piz-t+Pib+Pikm+Pi9 MR 1
Pia+Pikm+Pi9 S 1
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