Crops ›› 2019, Vol. 35 ›› Issue (3): 172-177.doi: 10.16035/j.issn.1001-7283.2019.03.029

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Molecular Detection and Analysis of Rice Blast Resistance Genes in Main Rice Varieties in Heilongjiang Province

Zhou Yili1,2,Zhang Yaling1,2,Zhao Hongsen1,2,Jin Xuehui1,2   

  1. 1 College of Agronomy, Heilongjiang Bayi Agricultural University, Daqing 163319, Heilongjiang, China
    2 Heilongjiang Plant Resistance Research Center, Daqing 163319, Heilongjiang, China
  • Received:2019-01-21 Revised:2019-03-31 Online:2019-06-15 Published:2019-06-12
  • Contact: Xuehui Jin

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Abstract:

Understanding the resistance genotype of the variety itself is great significant for the rational layout of rice varieties. In order to clarify the distribution of rice blast resistance genes Pi-ta, Pi-b, Pik-m, Pi9, Pii and Pi-d3 in rice varieties in Heilongjiang Province, 34 main varieties were selected, and the functional markers of these 6 anti-blast genes were used to detect the molecular markers. Results showed that the distribution frequency of Pi9 was the highest, followed by Pi-ta and Pik-m, the frequency of Pi-b and Pii resistance genes was lower, and the distribution frequency of Pi-d3 was the lowest. Among the 34 rice materials tested, some varieties contained up to five resistance genes and some contained at least one resistance gene. The species with two resistance genes accounted for the largest proportion. Longjing 40 and Longjing 42 did not contain the gene to be detected.

Key words: Rice, Resistance gene, Molecular detection, Distribution, Rice blast

Table 1

Primer sequence and amplified fragment size for PCR"

目的基因Target gene 引物名称Primer name 引物序列(5′→3′) Primer sequence (5′→3′) 预期片段大小Expected size (bp) 酶Enzyme
Pi-ta Pi-ta F
Pi-ta R		 AGCAGGTTATAAGCTAGGCC
CTACCAACAAGTTCATCAAA		 1042 -
Pi-ta Npi-ta F
Npi-ta R		 AGCAGGTTATAAGCTAGCTAT
CTACCAACAAGTTCATCAAA		 1042 -
Pi-b Pi-b F
Pi-b R		 GAACAATGCCCAAACTTGAG
GGGTCCACATGTCAGTGAGC		 365 -
Pi-b Npi-b F
Npi-b R		 TCGGTGCCTCGGTAGTCAGT
GGGAAGCGGATCCTAGGTCT		 803 -
Pik-m Pik-m1 F
Pik-m1 R		 TGAGCTCAAGGCAAGAGTTGAGGA
TGTTCCAGCAACTCGATGAG		 174（抗）/213 -
Pik-m Pik-m2 F
Pik-m2 R		 CAGTAGCTGTGTCTCAGAACTATG
AAGGTACCTCTTTTCGGCCAG		 290（抗）/332 -
Pi9 Pi9 F
Pi9 R		 GCTGTGCTCCAAATGAGGAT
GCGATCTCACATCCTTTGCT		 291（抗）/397 -
Pii Pii F
Pii R		 TCCAATGCTTCTGAAAGGTAGC
TGGAAACATGAACCCATATCCT		 355 PvuⅡ
Pi-d3 Pi-d3 F
Pi-d3 R		 TACTACTCATGGAAGCTAGTTCTC
ACGTCACAAATCATTCGCTC		 658 BamHⅠ

Fig.1

The results of Pi-ta, Pi-b gene amplification M, DL2 000; Lane 1-K1 (Pi-ta) in A, B; Lane 1-BL1 (Pi-b) in C, D; Lane 2-LTH; Lane 3-Lane 36 are 34 rice varieties selected"

Fig.2

The results of Pik-m, Pi9 gene amplification M, DL2 000; Lane 1-T suyuake (Pik-m) in A, B; lane 1-Oryza minuta (Pi9) in C; Lane 2-LTH; Lane 3-Lane 36 are 34 rice varieties selected"

Fig.3

The results of enzyme digestion of Pii and Pi-d3 M, DL2 000; Lane 1-Fujisaki 5 (Pii) PCR product in A, Lane 1-Digu (Pi-d3) PCR product in B; Lane 2-Fujisaki 5 (Pii) digestion product in A, Lane 2-Valley (Pi-d3) digestion product in B; Lane 3-LTH PCR product; Lane 4-LTH digestion product; Lane 5 to Lane 38 are 34 rice varieties selected"

Table 2

Results of molecular detection of resistant genes for 34 rice varieties"

编号
Numbering		 品种Variety 抗性基因Resistance gene
Pi-ta Pi-b Pik-m Pi9 Pii Pi-d3
1 龙粳21 Longjing 21 - - + + - -
2 龙粳26 Longjing 26 - - + - - -
3 龙粳29 Longjing 29 + - - + + -
4 龙粳31 Longjing 31 + - + - - -
5 龙粳40 Longjing 40 - - - - - -
6 龙粳42 Longjing 42 - - - - - -
7 龙粳43 Longjing 43 - + - + + -
8 龙粳45 Longjing 45 + - + + - -
9 龙粳46 Longjing 46 - - + + - -
10 龙粳50 Longjing 50 + - - + - -
11 龙粳52 Longjing 52 - - + + + -
12 龙粳53 Longjing 53 + - + + - -
13 松粳3 Songjing 3 + - - + + -
14 松粳15 Songjing 15 - + + + - -
15 松粳20 Songjing 20 + + + + + -
16 绥粳4 Suijing 4 + - - + - -
17 中龙粳1 Zhonglongjing 1 - + + + - -
18 龙洋1号 Longyang 1 + - + - - -
19 龙洋10 Longyang 10 - + - + - -
20 龙洋16 Longyang 16 + + - - + +
21 北稻3 Beidao 3 + - + + + -
22 北稻4 Beidao 4 + + + + - -
23 垦稻12 Kendao 12 - - + - - -
24 垦稻23 Kendao 23 - + - - - -
25 垦稻26 Kendao 26 + - - + - -
26 垦稻30 Kendao 30 + - - - - -
27 垦稻31 Kendao 31 + - + + - -
28 垦稻32 Kendao 32 + - + - - -
编号
Numbering		 品种Variety 抗性基因Resistance gene
Pi-ta Pi-b Pik-m Pi9 Pii Pi-d3
29 龙庆稻3号Longqingdao 3 + - - + - -
30 稻花香2号Daohuaxiang 2 + - - + + -
31 空育131 Kongyu 131 - - + + - -
32 稼禾1号Jiahe 1 + - - + - -
33 三江16 Sanjiang 16 - - - + - -
34 莎莎泥Shashani 16 - - + + - -
LTH - - - - - -
检出频率Detection frequency (%) 55.9 23.5 52.9 70.6 23.5 2.9
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