四川省丁香假单胞猕猴桃致病变种的遗传多样性分析

doi:10.11924/j.issn.1000-6850.casb18110082

摘要/Abstract

摘要：

为明确四川省猕猴桃溃疡病菌丁香假单胞菌猕猴桃致病变种(Pseudomonas syringae pv. actinidiae,Psa)的遗传多样性及群组划分与地理来源的相关性。以四川省不同地理来源的40个Psa菌株为研究试材,用rep-PCR技术进行分子标记,NTSYS软件分析UPGMA聚类。3对rep-PCR检测引物(REP、ERIC和BOX)共获得42个位点,其中38个为多态位点,占90.5%。UPGMA聚类结果显示,以相似系数为0.70阀值时,40个Psa菌株被分为2个类群(Ⅰ、Ⅱ),其中85.0%的菌株属于第Ⅱ类群;在相似性系数为0.80时,第Ⅰ类又可分为2个亚类(Ⅰ-1、Ⅰ-2),第Ⅱ类则分为5个亚类(Ⅱ-1、Ⅱ-2、Ⅱ-3、Ⅱ-4、Ⅱ-5),菌株无明显的采集地聚类。以上结果表明,四川省各地区的猕猴桃溃疡病菌菌株具有较高的遗传多样性,但其遗传变异与菌株地理来源无明显相关性。

关键词: 猕猴桃, 丁香假单胞菌猕猴桃致病变种, rep-PCR, 遗传多样性, 遗传变异

Abstract:

To determine the genetic diversity of kiwifruit ulcer pathogen Pseudomonas syringae pv. actinidiae (Psa) in Sichuan and the correlation between genetic group division and geographical origin, 40 Psa strains from different regions of Sichuan were used as the research materials, rep-PCR was used for molecular marker, and UPGMA clustering was conducted with NTSYS software. A total of 42 loci were obtained by 3 primer combinations (REP, ERIC and BOX), including 38 polymorphic loci, the rate of polymorphism was 90.5%. The results of UPMGA cluster analysis indicated that 40 Psa strains were divided into two groups (Ⅰ, Ⅱ) in the genetic similarity coefficient of 0.70, and 85.0% of the strains belonged to the Ⅱ group. The group Ⅰ contained two sub-groups (I-1, I-2) and the group Ⅱ consisted of five sub-groups (Ⅱ-1, Ⅱ-2, Ⅱ-3, Ⅱ-4, Ⅱ-5) in the genetic similarity coefficient of 0.80, but all strains had no obvious clustering of collecting locations. This study showed a rich polymorphism of Psa strains from different regions of Sichuan, but the genetic diversity had no significant correlation with geographical location.

Key words: kiwifruit, Pseudomonas syringae pv. actinidiae, rep-PCR, genetic diversity, genetic variation

中图分类号:

S432.1

杨潇湘, 伍文宪, 黄小琴, 张蕾, 杜磊, 刘勇. 四川省丁香假单胞猕猴桃致病变种的遗传多样性分析[J]. 中国农学通报, 2020, 36(9): 132-137.

Xiaoxiang Yang, Wenxian Wu, Xiaoqin Huang, Lei Zhang, Lei Du, Yong Liu. Genetic Diversity of Pseudomonas syringae pv. actinidiae strains in Sichuan[J]. Chinese Agricultural Science Bulletin, 2020, 36(9): 132-137.

图/表 6

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编号	名称	采集地	编号	名称	采集地	编号	名称	采集地
1	CX1	苍溪	15	QL3	邛崃	29	PZ2	彭州
2	CX2	苍溪	16	QL4	邛崃	30	PZ3	彭州
3	CX3	苍溪	17	PJ1	蒲江	31	PZ4	彭州
4	CX4	苍溪	18	PJ2	蒲江	32	PZ5	彭州
5	CX5	苍溪	19	PJ3	蒲江	33	PZ6	彭州
6	JY1	江油	20	PJ4	蒲江	34	PZ8	彭州
7	JY2	江油	21	PJ5	蒲江	35	PZ8	彭州
8	MN1	冕宁	22	PJ6	蒲江	36	SF1	什邡
9	MN2	冕宁	23	DJY1	都江堰	37	SF2	什邡
10	MS1	名山	24	DJY2	都江堰	38	SF3	什邡
11	MS2	名山	25	DJY3	都江堰	39	SF4	什邡
12	MS3	名山	26	DJY4	都江堰	40	SF5	什邡
13	QL1	邛崃	27	DJY5	都江堰
14	QL2	邛崃	28	PZ1	彭州

编号	名称	采集地	编号	名称	采集地	编号	名称	采集地
1	CX1	苍溪	15	QL3	邛崃	29	PZ2	彭州
2	CX2	苍溪	16	QL4	邛崃	30	PZ3	彭州
3	CX3	苍溪	17	PJ1	蒲江	31	PZ4	彭州
4	CX4	苍溪	18	PJ2	蒲江	32	PZ5	彭州
5	CX5	苍溪	19	PJ3	蒲江	33	PZ6	彭州
6	JY1	江油	20	PJ4	蒲江	34	PZ8	彭州
7	JY2	江油	21	PJ5	蒲江	35	PZ8	彭州
8	MN1	冕宁	22	PJ6	蒲江	36	SF1	什邡
9	MN2	冕宁	23	DJY1	都江堰	37	SF2	什邡
10	MS1	名山	24	DJY2	都江堰	38	SF3	什邡
11	MS2	名山	25	DJY3	都江堰	39	SF4	什邡
12	MS3	名山	26	DJY4	都江堰	40	SF5	什邡
13	QL1	邛崃	27	DJY5	都江堰
14	QL2	邛崃	28	PZ1	彭州

引物名称	引物序列(5'--3')
REP1R-I	IIIICGICGICATCIGGC
REP2-I	ICGICTTATCIGGCCTAC
ERIC1R	ATGTAAGCTCCTGGGGATTCAC
ERIC2	AAGTAAGTGACTGGGGTGAGCG
BOXA1R	CTACGGCAAGGCGACGCTGACG
PsaF1	TTTTGCTTTGCACACCCGATTTT
PsaR2	CACGCACCCTTCAATCAGGATG

引物名称	引物序列(5'--3')
REP1R-I	IIIICGICGICATCIGGC
REP2-I	ICGICTTATCIGGCCTAC
ERIC1R	ATGTAAGCTCCTGGGGATTCAC
ERIC2	AAGTAAGTGACTGGGGTGAGCG
BOXA1R	CTACGGCAAGGCGACGCTGACG
PsaF1	TTTTGCTTTGCACACCCGATTTT
PsaR2	CACGCACCCTTCAATCAGGATG

引物	位点总数	多态性位点数	多态性比率/%
REP1R-I/REP2-I	10	9	90.0
ERIC1/ERIC2	16	15	93.8
BOXA1R	16	14	87.5
总数	42	38	90.5
平均数	14	12.7	90.5