Crops ›› 2018, Vol. 34 ›› Issue (3): 168-173.doi: 10.16035/j.issn.1001-7283.2018.03.026

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Genetic Diversity of Phytophthora infestans in Northeast China

Tian Huiyao,Jiang Jizhi,Li Chengbin,Shen Fen,Hou Ning   

  1. College of Life Sciences, Hebei University, Baoding 071002, Hebei, China
  • Received:2017-12-25 Revised:2018-05-08 Online:2018-06-20 Published:2018-06-20

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Abstract:

In order to know the genetic differences at DNA level among Phytophthora infestans isolates collected from Northeast China in 2016, the polymorphisms of 117 isolates were detected by two loci, Pi4B and Pi4G of SSR primers, and nominated based on SSR molecular marker. The results showed that 10 allele genes and 14 SSR genotypes were identified, among which allele genes 161 and 206 accounted for significant advantages. A total of 14 kinds of SSR genotypes were A-03, A-10, B-01, D-03, E-01, F-01, F-02, F-06, G-02, G-03, G-07, K-01, L-01, and N-01, respectively. A-10 and N-01 were novel genotypes that had not existed in this regions, and F-06 was the dominant genotype, accounting for 29.0% in the measure isolates. At the genetic coefficient of 0.85, 14 kinds of SSR genotypes varieties were divided into 10 groups by UPGMA cluster analysis. There was no correlation between population genetic diversity of P. infestans and geographical origin. Also, through population genetic diversity analysis by using POPGENE1.32 software, genetic difference was small among all tested P. infestans populations from Heilongjiang, Jilin, and Liaoning. The minimum genetic distance was only 0.0213 between Heilongjiang Province and Jilin Province. These findings indicated that SSR genotypes of P. infestans in Northeast China had many groups, and its genetic diversity was abundant.

Key words: Phytophthora infestans, DNA fingerprinting, SSR genotype, Genetic diversity

Table 1

Sample number of P.infestans isolated and identified from Heilongjiang, Jilin and Liaoning in 2016"

采集地点
Sample site		 黑龙江Heilongjiang 吉林Jilin 辽宁Liaoning 合计
Total
双城区(HS) 青冈县(HQ) 梨树(JL) 敦化(JD) 新民(LX) 昌图(LC)
病叶数Number of diseased leaf 5 5 5 5 5 5 30
菌株数Number of strain 18 22 16 21 21 19 117

Fig.1

Agarose gel electrophoresis of genomic DNA of P.infestans strains"

Table 2

Distribution and frequency of SSR genotype of P. infestans in Heilongjiang, Jilin and Liaoning"

基因型
Genotype		 采样地点Sampling site 合计
Total		 百分比(%)
Percentage
黑龙江
Heilongjiang		 吉林
Jilin		 辽宁
Liaoning
A-03 1 2 2 5 4.3
A-10 2 2 1 5 4.3
B-01 1 2 4 7 5.9
D-03 3 3 4 10 8.6
E-01 2 1 - 3 2.3
F-01 6 8 6 20 17.3
F-02 2 3 4 9 7.6
F-06 13 11 10 34 29.0
G-02 2 - - 2 1.8
G-03 4 3 1 8 6.9
G-07 1 1 2 4 3.6
K-01 2 - 1 3 2.3
L-01 1 1 3 5 4.3
N-01 - - 2 2 1.8
合计 40 37 40 117 100.0

Table 3

SSR new genotypes of P.infestans strains based on primers Pi4B and Pi4G"

基因型Genotype Pi4G Pi4B
161bp 163bp 165bp 177bp 179bp 206bp 214bp 218bp 220bp
*A-10 √ — — — √ — — √ —
*N-01 — √ — — √ √ — √ —

Fig.2

Molecular phylogenetic tree of P.infestans according to SSR genotypes by UPGMA method"

Table 4

Strains of different SSR types"

SSR类型SSR genotype 菌株名称Strain name
A-03 HS-01、LX-19、JL-23
A-10 HS-05、LX-23
D-03 HS-10、HS-17、JL-19、JD-21
A-01 DB-606
F-01 HQ-02、LC-03、JL-02、JD-09
F-02 HQ-09、LC-21、JD-13、JD-07
G-02 HQ-10
E-01 JL-05
L-01 LX-06
B-01 HS-07、LC-05
G-07 JL-19
F-06 HQ-05、LC-18
N-01 LX-04
G-03 HQ-14、JD-02
K-01 LX-08

Table 5

Alleles frequencies and diversity of P. infestans population in two microsatellite loci"

基因座
Locus		 等位
基因
Allele		 等位基因频率Allele frequency Shannon指数
Shannon index
黑龙江
Heilongjiang		 吉林
Jilin		 辽宁
Liaoning
Pi4G 159a 0.0729 0.0000 0.0000 0.7812
161c 0.5132 0.5926 0.4054
163c 0.3353 0,3008 0.3714
165a 0.0000 0.0000 0.1224
177b 0.0717 0.0455 0.0000
179c 0.0706 0.0911 0.1531
Pi4B 206c 0.4282 0.4987 0.4216 0.9325
214c 0.1345 0.0456 0.0413
218c 0.3011 0.3212 0.2913
220b 0.0938 0.0000 0.1479
平均数Average 0.8569

Table 6

Genetic distance and genetic similarity of P. infestans collected from Heilongjiang, Jilin and Liaoning"

采样地点Sampling site 黑龙江Heilongjiang 吉林Jilin 辽宁Liaoning
黑龙江Heilongjiang 0.9811 0.9165
吉林Jilin 0.0213 0.9439
辽宁Liaoning 0.0847 0.0533
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