利用SRAP分子标记分析谷瘟病菌遗传多样性

doi:10.7668/hbnxb.20194152

摘要/Abstract

摘要：

了解谷瘟病菌的变异和群体结构,为今后谷瘟病的防控及抗病品种培育提供理论基础。利用20对SRAP引物对采自9个地区的90株谷瘟病菌菌株进行了PCR扩增,利用NTSYSpc-2.11F软件进行数据分析,通过UPGMA方法进行聚类分析,使用Popgene 32软件计算各群体间的遗传多样性指数。结果表明,筛选出了8对引物用于谷瘟病菌的遗传多态性分析,这8对引物共扩增出条带1 728条,其中多态性条带1 492条,多态性比率为86.34%。对90株病原菌进行聚类分析,其相似性系数为0.77~0.85,遗传相似系数为0.802时,所有菌株被分为27个遗传宗谱(L1~L27),其中L1为绝对优势组群,共包含来自山东、河北、山西、河南和辽宁5个省份的29株谷瘟病菌,占总菌株的32.22%。经Popgene 32软件计算分析,9个地区谷瘟病菌的Nei's遗传多样性指数(H)为0.141 4~0.288 1,Shannon's信息指数(I)为0.196 0~0.441 6,河北夏谷区Nei's遗传多样性指数和Shannon's信息指数最高,遗传多样性最丰富,而海南群体遗传多样性最低。对不同地区谷瘟病菌群体比较,吉林群体与海南群体的遗传亲缘关系最远,而河北夏谷区群体与河北春谷区群体的亲缘关系最近。可见,不同地区谷瘟病菌的遗传多样性丰富,各菌株间存在遗传分化,但菌株间的遗传分化与地理来源无明显相关性。

关键词: 谷子, 谷瘟病菌, SRAP, 相似性系数, 遗传多样性

Abstract:

Understanding the variation and population structure of the foxtail millet blast provides a theoretical basis for future cultivation of resistant varieties and control of the disease.Twenty pairs of SRAP primers were used for PCR amplification of strains of P.oryzae collected from 9 regions.NTSYSpc-2.11F software was used for data analysis,UPGMA method was used for cluster analysis,and Popgene 32 software was used to calculate the genetic diversity index among populations.The results showed that 8 pairs of primers were selected with good polymorphism.A total of 1 728 discernible bands were amplified by 8 pairs of primers,1 492 of which showed polymorphism,and polymorphism rate was 86.34%.The cluster analysis of 90 strains of P.oryzae showed that the similarity coefficient was between 0.77-0.85.All strains were divided into 27 genetic lineages (L1-L27) when the genetic similarity coefficient was 0.802.Among them,L1 was the absolute dominant group,containing 29 strains of P.oryzae,which from Shandong,Hebei,Shanxi,Henan and Liaoning Province,accounting for 32.22% of the total strains.According to the calculation and analysis of Popgene 32 software,Nei's genetic diversity index (H) and Shannon's information index (I) of foxtail millet blast in 9 regions ranged from 0.141 4 to 0.288 1,and 0.196 0 to 0.441 6,respectively.Nei's genetic diversity index and Shannon's information index in the summer sowing region of Hebei Province were the highest,with the richest genetic diversity,while Hainan population had the lowest genetic diversity.Compared with different regions of the populations,the genetic relationship between the Jilin population and the Hainan population was the farthest,while the genetic relationship between the Hebei summer sowing millet region population and the Hebei spring sowing millet region population was the closest.It can be seen that the genetic diversity of P.oryzae in different regions was rich,and there was genetic differentiation among strains,but there was no significant correlation between genetic differentiation and geographical origin.

Key words: Foxtail millet, Pyricularia oryzae, SRAP, Similarity coefficient, Genetic diversity

刘佳, 张梦雅, 任世龙, 王永芳, 马继芳, 全建章, 刘磊, 董志平, 白辉, 李志勇. 利用SRAP分子标记分析谷瘟病菌遗传多样性[J]. 华北农学报, 2023, 38(5): 179-187. doi: 10.7668/hbnxb.20194152.

LIU Jia, ZHANG Mengya, REN Shilong, WANG Yongfang, MA Jifang, QUAN Jianzhang, LIU Lei, DONG Zhiping, BAI Hui, LI Zhiyong. Genetic Diversity of Pyricularia oryzae Based on SRAP Molecular Markers[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(5): 179-187. doi: 10.7668/hbnxb.20194152.

http://www.hbnxb.net/CN/Y2023/V38/I5/179

图/表 8

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编号 Number	菌株名称 Isolate code	来源 Origin	编号 Number	菌株名称 Isolate code	来源 Origin
1~13	CD1~CD13	河北春谷,承德	60	SD6	山东,济南
14~20	HB1~HB7	河北夏谷,石家庄	61~62	SD7~SD8	山东,临沂
21~28	HB8~HB15	河北夏谷,邯郸	63~66	SX1~SX4	山西,长治
29~32	HB16~HB19	河北夏谷,保定	67~68	SX5~SX6	山西,晋中
33~34	HB20~HB21	河北夏谷,沧州	69~71	SX7~SX9	山西,大同
35~43	HB22~HB30	河北夏谷,邢台	72~73	LN1~LN2	辽宁,朝阳
44~45	HB31~HB32	河北夏谷,衡水	74~79	LN3~LN8	辽宁,北票
46~48	HN1~HN3	河南,原阳	80~81	JL1~JL2	吉林
49	HN4	河南,郑州	82~84	HLJ1~HLJ3	黑龙江,哈尔滨
50	HN5	河南,新乡	85~86	HLJ4~HLJ5	黑龙江,齐齐哈尔
51~54	HN6~HN9	河南,安阳	87~90	Hn1~Hn4	海南,三亚
55~59	SD1~SD5	山东,潍坊

编号 Number	菌株名称 Isolate code	来源 Origin	编号 Number	菌株名称 Isolate code	来源 Origin
1~13	CD1~CD13	河北春谷,承德	60	SD6	山东,济南
14~20	HB1~HB7	河北夏谷,石家庄	61~62	SD7~SD8	山东,临沂
21~28	HB8~HB15	河北夏谷,邯郸	63~66	SX1~SX4	山西,长治
29~32	HB16~HB19	河北夏谷,保定	67~68	SX5~SX6	山西,晋中
33~34	HB20~HB21	河北夏谷,沧州	69~71	SX7~SX9	山西,大同
35~43	HB22~HB30	河北夏谷,邢台	72~73	LN1~LN2	辽宁,朝阳
44~45	HB31~HB32	河北夏谷,衡水	74~79	LN3~LN8	辽宁,北票
46~48	HN1~HN3	河南,原阳	80~81	JL1~JL2	吉林
49	HN4	河南,郑州	82~84	HLJ1~HLJ3	黑龙江,哈尔滨
50	HN5	河南,新乡	85~86	HLJ4~HLJ5	黑龙江,齐齐哈尔
51~54	HN6~HN9	河南,安阳	87~90	Hn1~Hn4	海南,三亚
55~59	SD1~SD5	山东,潍坊

上游引物名称 Forward primer	引物序列(5'-3') Primer sequence(5'-3')	下游引物名称 Reverse primer	引物序列(5'-3') Primer sequence(5'-3')
Me4	TGAGTCCAAACCGGACC	Em1	GACTGCGTACGAATTAAT
Me5	TGAGTCCAAACCGGAAG	Em2	GACTGCGTACGAATTTGC
Me6	TGAGTCCAAACCGGTAA	Em4	GACTGCGTACGAATTTGA
Me7	TGAGTCCAAACCGGACA	Em7	GACTGCGTACGAATTCAA
Me8	TGAGTCCAAACCGGTGT

上游引物名称 Forward primer	引物序列(5'-3') Primer sequence(5'-3')	下游引物名称 Reverse primer	引物序列(5'-3') Primer sequence(5'-3')
Me4	TGAGTCCAAACCGGACC	Em1	GACTGCGTACGAATTAAT
Me5	TGAGTCCAAACCGGAAG	Em2	GACTGCGTACGAATTTGC
Me6	TGAGTCCAAACCGGTAA	Em4	GACTGCGTACGAATTTGA
Me7	TGAGTCCAAACCGGACA	Em7	GACTGCGTACGAATTCAA
Me8	TGAGTCCAAACCGGTGT

引物组合 Primer combination	总条带数/条 Total of band	多态性条带数目/条 Polymorphic band	多态性比率/% Percentage of polymorphic band
Me4/Em7	216	172	79.63
Me5/Em2	216	180	83.33
Me6/Em2	216	170	78.70
Me6/Em4	216	180	83.33
Me6/Em7	216	207	95.83
Me7/Em1	216	204	94.44
Me8/Em2	216	188	87.04
Me8/Em7	216	191	88.43
总计Total	1 728	1 492	86.34

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