海南南繁区水稻纹枯病菌的遗传多样性与致病力分化

doi:10.16819/j.1001-7216.2019.8048

摘要/Abstract

摘要：

【目的】为明确南繁区水稻纹枯病菌(Rhizoctonia solani AG-1 IA)的遗传分化及遗传多样性与致病力的关系,【方法】采用AFLP分子标记技术和离体叶片接种法对南繁核心区(三亚、乐东、陵水)和非核心区(琼中、屯昌和定安)共60株水稻纹枯病菌的遗传多样性、遗传结构和致病力分化进行了测定,并分析了遗传多样性与致病力之间的关系。【结果】聚类分析结果表明,核心区菌株的遗传多样性相对更高;群体遗传结构分析表明,核心区群体的多态性位点百分率(PPL)、Nei基因多样性指数(H)、Shannon信息指数(I)和基因流(N_m)分别为 82.24%、0.1932、0.3062和2.5627,高于非核心区群体的67.49%、0.1535、0.2447和0.9365;而核心区群体的基因分化系数(G_st)为0.1633,低于非核心区群体的0.3481。【结论】核心区菌株的遗传变异程度比非核心区菌株高;核心区不同群体间存在较多的基因交流,而非核心区菌体间的基因交流较少,但遗传变异均主要来自于群体内;总体而言,核心区菌株的遗传分化程度更高。菌株的致病力及其与菌株遗传多样性的相关性分析表明,核心区菌株以中等致病型为主,而非核心区菌株则以中、强致病型为主,但与菌株的AFLP谱系之间的相关均未达显著水平。

关键词: 南繁区, 水稻纹枯病菌, 遗传多样性, 遗传结构, 致病力分化, AFLP

Abstract:

【Objective】In order to clarify the genetic differentiation and the correlation between genetic diversity and pathogenicity of Rhizoctonia solani AG-1 ⅠA in South China Crop Breeding Area, the genetic diversity, population genetic structure, pathogenicity and the correlation between genetic diversity and pathogenicity of 60 R. solani AG-1 IA isolates collected from the core region (Sanya, Ledong and Lingshui) and non-core region (Qiongzhong, Tunchang and Ding’an) of South China Crop Breeding Area were comparatively analyzed 【Method】by using AFLP molecular marker technique and detached leaf inoculation method. 【Result】The cluster analysis results showed that the genetic diversity of R. solani AG-1 ⅠA in the core region was relatively higher. The analysis results of population genetic structure showed that the percentage of polymorphic loci (PPL), Nei’s gene diversity index (H), Shannon’s information index (I) and gene flow (N_m) in the core population were 82.24%, 0.1932, 0.3062 and 2.5627, respectively, higher than those of non-core population with PPL, H, I and N_m being 67.49%, 0.1535, 0.2447 and 0.9365, respectively. However, the genetic differentiation coefficient (G_st) in the core population was 0.1633, lower than that of non-core population whose G_st was 0.3481. 【Conclusion】These results showed that the degree of genetic variation of the isolates from core region was higher than that of the isolates from non-core region. Much gene flow among the core populations and little gene flow among the non-core populations existed, but the genetic variation of both regions was mainly within the population. Overall, the degree of genetic differentiation in the isolates of core region was higher. The results of pathogenicity and correlation between genetic diversity and pathogenicity showed that most isolates from core region belonged to the medium pathotypes and most isolates from non-core region belonged to the medium or strong pathotypes, but there was no correlation between pathogenicity and AFLP lineage.

Key words: South China Crop Breeding Area, Rhizoctonia solani AG-1 ⅠA;, genetic diversity, population genetic structure, pathogenicity differentiation, AFLP

朱名海, 彭丹丹, 舒灿伟, 周而勋. 海南南繁区水稻纹枯病菌的遗传多样性与致病力分化[J]. 中国水稻科学, 2019, 33(2): 176-185.

Minghai ZHU, Dandan PENG, Canwei SHU, Erxun ZHOU. Genetic Diversity and Pathogenicity Differentiation of Rhizoctonia solani AG-1 IA from South China Crop Breeding Area in Hainan Province[J]. Chinese Journal OF Rice Science, 2019, 33(2): 176-185.

图/表 12

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区域 Region	采集地点 Sampling location	采集时间 Sampling date	菌株编号 Isolate No.
核心区Core region	三亚Sanya	2014-05-28	SY1, SY2, SY3, SY4, SY5, SY6, SY7, SY8, SY9, SY10
	乐东Ledong	2014-06-01	LD1, LD2, LD3, LD4, LD5, LD6, LD7, LD8, LD9, LD10
	陵水Lingshui	2014-03-15, 2014-10-27	LS1, LS2, LS3, LS4, LS5, LS6, LS7, LS8, LS8, LS10
非核心区Non-core region	琼中Qiongzhong	2014-06-02	QZ1, QZ2, QZ3, QZ4, QZ5, QZ6, QZ7, QZ8, QZ9, QZ10
	屯昌Tunchang	2014-06-03	TC1, TC2, TC3, TC4, TC5, TC6, TC7, TC8, TC9, TC10
	定安Ding’an	2014-06-04	DA1, DA2, DA3, DA4, DA5, DA6, DA7, DA8, DA9, DA10

区域 Region	采集地点 Sampling location	采集时间 Sampling date	菌株编号 Isolate No.
核心区Core region	三亚Sanya	2014-05-28	SY1, SY2, SY3, SY4, SY5, SY6, SY7, SY8, SY9, SY10
	乐东Ledong	2014-06-01	LD1, LD2, LD3, LD4, LD5, LD6, LD7, LD8, LD9, LD10
	陵水Lingshui	2014-03-15, 2014-10-27	LS1, LS2, LS3, LS4, LS5, LS6, LS7, LS8, LS8, LS10
非核心区Non-core region	琼中Qiongzhong	2014-06-02	QZ1, QZ2, QZ3, QZ4, QZ5, QZ6, QZ7, QZ8, QZ9, QZ10
	屯昌Tunchang	2014-06-03	TC1, TC2, TC3, TC4, TC5, TC6, TC7, TC8, TC9, TC10
	定安Ding’an	2014-06-04	DA1, DA2, DA3, DA4, DA5, DA6, DA7, DA8, DA9, DA10

序号 Code	接头或引物 Adapter or primer	序列 Primer sequence(5′→3′)	引物用途 Purpose
Ea	EcoRⅠ接头 EcoRⅠadapter	CTCGTAGACTGCGTACC CATCTGACGCATGGTTAA	连接 Ligation
Em	MseⅠ接头 MseⅠadapter	GACGATGAGTCCTGAG TACTCAGGACTCAT	连接 Ligation
E0	EcoRⅠ+0	GACTGCGTACCAATTC	预扩增 Pre-amplification
E1	EcoRⅠ+AA	GACTGCGTACCAATTCAA	选择性扩增 Selective amplification
E2	EcoRⅠ+AT	GACTGCGTACCAATTCAT	选择性扩增 Selective amplification
E3	EcoRⅠ+AC	GACTGCGTACCAATTCAC	选择性扩增 Selective amplification
E4	EcoRⅠ+AG	GACTGCGTACCAATTCAG	选择性扩增 Selective amplification
E5	EcoRⅠ+TA	GACTGCGTACCAATTCTA	选择性扩增 Selective amplification
E6	EcoRⅠ+TT	GACTGCGTACCAATTCTT	选择性扩增 Selective amplification
E7	EcoRⅠ+TC	GACTGCGTACCAATTCTC	选择性扩增 Selective amplification
E8	EcoRⅠ+TG	GACTGCGTACCAATTCTG	选择性扩增 Selective amplification
M0	MseⅠ+0	GATGAGTCCTGAGTAA	预扩增 Pre-amplification
M1	MseⅠ+CAA	GATGAGTCCTGAGTAACAA	选择性扩增 Selective amplification
M2	MseⅠ+CAT	GATGAGTCCTGAGTAACAT	选择性扩增 Selective amplification
M3	MseⅠ+CAC	GATGAGTCCTGAGTAACAC	选择性扩增 Selective amplification
M4	MseⅠ+CAG	GATGAGTCCTGAGTAACAG	选择性扩增 Selective amplification
M5	MseⅠ+ CTA	GATGAGTCCTGAGTAACTA	选择性扩增 Selective amplification
M6	MseⅠ+CTT	GATGAGTCCTGAGTAACTT	选择性扩增 Selective amplification
M7	MseⅠ+CTC	GATGAGTCCTGAGTAACTC	选择性扩增 Selective amplification
M8	MseⅠ+CTG	GATGAGTCCTGAGTAACTG	选择性扩增 Selective amplification

序号 Code	接头或引物 Adapter or primer	序列 Primer sequence(5′→3′)	引物用途 Purpose
Ea	EcoRⅠ接头 EcoRⅠadapter	CTCGTAGACTGCGTACC CATCTGACGCATGGTTAA	连接 Ligation
Em	MseⅠ接头 MseⅠadapter	GACGATGAGTCCTGAG TACTCAGGACTCAT	连接 Ligation
E0	EcoRⅠ+0	GACTGCGTACCAATTC	预扩增 Pre-amplification
E1	EcoRⅠ+AA	GACTGCGTACCAATTCAA	选择性扩增 Selective amplification
E2	EcoRⅠ+AT	GACTGCGTACCAATTCAT	选择性扩增 Selective amplification
E3	EcoRⅠ+AC	GACTGCGTACCAATTCAC	选择性扩增 Selective amplification
E4	EcoRⅠ+AG	GACTGCGTACCAATTCAG	选择性扩增 Selective amplification
E5	EcoRⅠ+TA	GACTGCGTACCAATTCTA	选择性扩增 Selective amplification
E6	EcoRⅠ+TT	GACTGCGTACCAATTCTT	选择性扩增 Selective amplification
E7	EcoRⅠ+TC	GACTGCGTACCAATTCTC	选择性扩增 Selective amplification
E8	EcoRⅠ+TG	GACTGCGTACCAATTCTG	选择性扩增 Selective amplification
M0	MseⅠ+0	GATGAGTCCTGAGTAA	预扩增 Pre-amplification
M1	MseⅠ+CAA	GATGAGTCCTGAGTAACAA	选择性扩增 Selective amplification
M2	MseⅠ+CAT	GATGAGTCCTGAGTAACAT	选择性扩增 Selective amplification
M3	MseⅠ+CAC	GATGAGTCCTGAGTAACAC	选择性扩增 Selective amplification
M4	MseⅠ+CAG	GATGAGTCCTGAGTAACAG	选择性扩增 Selective amplification
M5	MseⅠ+ CTA	GATGAGTCCTGAGTAACTA	选择性扩增 Selective amplification
M6	MseⅠ+CTT	GATGAGTCCTGAGTAACTT	选择性扩增 Selective amplification
M7	MseⅠ+CTC	GATGAGTCCTGAGTAACTC	选择性扩增 Selective amplification
M8	MseⅠ+CTG	GATGAGTCCTGAGTAACTG	选择性扩增 Selective amplification

引物组合 Primer combination	扩增条带数 Amplification band number	多态性条带数 Polymorphic band number	多态性条带百分率 Percentage of polymorphic bands/%
E1/M2	63	60	95.24
E2/M3	39	34	87.18
E2/M7	44	33	75.00
E3/M2	57	48	84.21
E3/M4	38	37	97.37
E6/M3	52	49	94.23
E6/M5	38	37	97.37
E6/M8	35	33	94.29
合计Total	366	331	90.44