Crops ›› 2021, Vol. 37 ›› Issue (4): 59-66.doi: 10.16035/j.issn.1001-7283.2021.04.009

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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 E-mail:gqing1198@163.com;Jxh2686@163.com

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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

Fig.1

Sketch map of sampling sites of tested strains"

Table 1

Primer information for specific molecular markers"

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

Table 2

Resistance frequency and resistance evaluation of the tested rice varieties"

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

Fig.2

The amplification results of rice blast resistance genes in some rice varieties M-DL 2000 DNA marker. 1-Disease resistance control materials for each gene; 2- Sensitive control material (LTH); 3-Longjing 21; 4-Longjing 26; 5-Longjing 29; 6-Longjing 31; 7-Longjing 40; 8-Longjing 42; 9-Longjing 43; 10-Kendao 12; 11-Kongyu 131; 12-Suijing 4"

Table 3

The results of molecular marker detection of resistance genes in tested rice varieties"

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

Fig.3

The distribution of resistance genes in different types of japonica rice varieties"

Table 4

Number of target genes and its ratio of japonica rice varieties"

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

Table 5

Rice blast resistance statistics of different resistant genotypes"

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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