南繁区稻瘟病菌遗传多样性和群体遗传结构的AFLP分析

doi:10.16819/j.1001-7216.2017.6136 320

摘要/Abstract

摘要：

【目的】为了明确南繁区稻瘟病菌(Magnaporthe oryzae)的遗传分化情况,【方法】采用AFLP分子标记技术对南繁核心区(三亚、乐东和保亭)和非核心区(琼中、屯昌和定安)共60个稻瘟病菌菌株的遗传多样性和群体遗传结构进行了比较分析。【结果】聚类分析表明,几乎所有菌株都聚在同一个谱系里,并且该谱系没有明显的亚群;群体遗传结构分析表明,核心区群体的多态性位点百分率、Shannon信息指数和基因流分别为 87.89%、0.2738和4.2897,高于非核心区群体的 81.37%、0.2703和3.5892;然而,核心区群体的Nei基因多样性指数和基因分化系数分别为 0.1657和0.1044,低于非核心区群体的 0.1662和0.1223。【结论】这些结果表明核心区和非核心区菌株都存在丰富的遗传多样性,不同群体间均存在较多的基因交流,但遗传变异均主要来自群体内;相比之下,核心区菌株的遗传多样性和遗传分化程度较高。

关键词: 南繁区, 稻瘟病菌, 遗传多样性, 群体遗传结构, AFLP

Abstract:

【Objective】 In order to clarify the genetic differentiation of Magnaporthe oryzae in South China Crop Breeding Area in Hainan Province, 【Method】the genetic diversity and population genetic structure of 60 Magnaporthe oryzae isolates collected from the core region (Sanya, Ledong and Baoting) and non-core region (Qiongzhong, Tunchang and Ding’an) of South China Crop Breeding Area were comparatively analyzed using amplified fragment length polymorphism(AFLP) technique. 【Result】The cluster analysis showed that almost all isolates were clustered in one group, and there was no obvious subgroup. The analysis of population genetic structure showed that the percentage of polymorphic loci(PPL), Shannon’s information index(I) and gene flow(N_m) in the core population were 87.89%, 0.2738 and 4.2897, respectively, higher than those of non-core population whose PPL, I and Nm were 81.37%, 0.2703 and 3.5892, respectively. However, Nei’s gene diversity index(H) and genetic differentiation coefficient(G_st) in the core population were 0.1657 and 0.1044, lower than those of non-core population whose H and G_st was 0.1662 and 0.1223. 【Conclusion】 These results showed that there were rich genetic diversity in both isolates of core and non-core regions and wide gene flow existed among these populations, but the genetic variation was mainly within the population. In contrast, the genetic diversity and the degree of genetic differentiation in the isolates of core region were relatively larger.

Key words: South China Crop Breeding Area, Magnaporthe oryzae, genetic diversity, population genetic structure, AFLP

中图分类号:

S435.111.4⁺1

朱名海, 皮磊, 舒灿伟, 周而勋. 南繁区稻瘟病菌遗传多样性和群体遗传结构的AFLP分析[J]. 中国水稻科学, 2017, 31(3): 320-326.

Minghai ZHU, Lei PI, Canwei SHU, Erxun ZHOU. AFLP Analyses of Genetic Diversity and Population Genetic Structure of Magnaporthe oryzae from South China Crop Breeding Area[J]. Chinese Journal OF Rice Science, 2017, 31(3): 320-326.

图/表 9

参考文献 19

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区域 Region	采集地点 Sampling location
南繁核心区Core region
三亚Sanya	育才镇国营立才农场	P1, P2, P3	2015-05-20	籼稻 indica
	吉阳镇大茅村	P4, P5, P6	2014-10-24	籼稻 indica
	吉阳镇罗蓬村	P7, P8	2014-10-24	籼稻 indica
	育才镇雅亮村	P9, P10	2014-10-29	籼稻 indica
乐东Ledong	抱由镇城郊	P11, P12, P13	2014-10-22	籼稻 indica
	利国镇新丰田洋	P14, P15, P16	2014-10-20	籼稻 indica
	千家镇农技实验站	P17, P18, P19, P20	2014-10-20	籼稻 indica
保亭Baoting	加茂镇灶长村	P21, P22, P23, P24, P25, P26, P27, P28, P29, P30	2015-05-15	籼稻 indica
南繁非核心区Non-core region
琼中Qiongzhong	长征镇长征农场	P31, P32, P33, P34, P35, P36, P37, P38, P39, P40	2015-05-18	籼稻 indica
屯昌Tunchang	屯城镇良史村	P41, P42, P43, P44, P45, P46, P47, P48, P49, P50	2015-05-18	籼稻 indica
定安Ding’an	新竹镇新序村	P51, P52, P53, P54, P55, P56, P57, P58, P59, P60	2015-05-17	籼稻 indica

区域 Region	采集地点 Sampling location
南繁核心区Core region
三亚Sanya	育才镇国营立才农场	P1, P2, P3	2015-05-20	籼稻 indica
	吉阳镇大茅村	P4, P5, P6	2014-10-24	籼稻 indica
	吉阳镇罗蓬村	P7, P8	2014-10-24	籼稻 indica
	育才镇雅亮村	P9, P10	2014-10-29	籼稻 indica
乐东Ledong	抱由镇城郊	P11, P12, P13	2014-10-22	籼稻 indica
	利国镇新丰田洋	P14, P15, P16	2014-10-20	籼稻 indica
	千家镇农技实验站	P17, P18, P19, P20	2014-10-20	籼稻 indica
保亭Baoting	加茂镇灶长村	P21, P22, P23, P24, P25, P26, P27, P28, P29, P30	2015-05-15	籼稻 indica
南繁非核心区Non-core region
琼中Qiongzhong	长征镇长征农场	P31, P32, P33, P34, P35, P36, P37, P38, P39, P40	2015-05-18	籼稻 indica
屯昌Tunchang	屯城镇良史村	P41, P42, P43, P44, P45, P46, P47, P48, P49, P50	2015-05-18	籼稻 indica
定安Ding’an	新竹镇新序村	P51, P52, P53, P54, P55, P56, P57, P58, P59, P60	2015-05-17	籼稻 indica

序号 Code	接头名称 Adapter name	引物序列 Primer sequence(5′→3′)
Ea	EcoRⅠ接头 EcoRⅠadapter	CTCGTAGACTGCGTACC CATCTGACGCATGGTTAA	连接 Connection
Em	MseⅠ接头 MseⅠadapter	GACGATGAGTCCTGAG TACTCAGGACTCAT	连接 Connection
E0	EcoRⅠ+0	GACTGCGTACCAATTC	预扩增 Pre-amplification
E1	EcoRⅠ+AAC	GACTGCGTACCAATTCAAC	选择性扩增Selective amplification analysis
E2	EcoRⅠ+ACA	GACTGCGTACCAATTCACA	选择性扩增 Selective amplification analysis
E3	EcoRⅠ+ACG	GACTGCGTACCAATTCACG	选择性扩增 Selective amplification analysis
E4	EcoRⅠ+AGG	GACTGCGTACCAATTCAGG	选择性扩增 Selective amplification analysis
E5	EcoRⅠ+AAG	GACTGCGTACCAATTCAAG	选择性扩增 Selective amplification analysis
E6	EcoRⅠ+ACT	GACTGCGTACCAATTCACT	选择性扩增 Selective amplification analysis
E7	EcoRⅠ+ACC	GACTGCGTACCAATTCACC	选择性扩增 Selective amplification analysis
E8	EcoRⅠ+AGC	GACTGCGTACCAATTCAGC	选择性扩增 Selective amplification analysis
M0	MseⅠ+0	GATGAGTCCTGAGTAA	预扩增预扩增 Pre-amplification
M1	MseⅠ+CAA	GATGAGTCCTGAGTAACAA	选择性扩增 Selective amplification analysis
M2	MseⅠ+CAT	GATGAGTCCTGAGTAACAT	选择性扩增 Selective amplification analysis
M3	MseⅠ+CAC	GATGAGTCCTGAGTAACAC	选择性扩增 Selective amplification analysis
M4	MseⅠ+CAG	GATGAGTCCTGAGTAACAG	选择性扩增 Selective amplification analysis
M5	MseⅠ+ CTA	GATGAGTCCTGAGTAACTA	选择性扩增 Selective amplification analysis
M6	MseⅠ+CTT	GATGAGTCCTGAGTAACTT	选择性扩增 Selective amplification analysis
M7	MseⅠ+CTC	GATGAGTCCTGAGTAACTC	选择性扩增 Selective amplification analysis
M8	MseⅠ+CTG	GATGAGTCCTGAGTAACTG	选择性扩增 Selective amplification analysis

序号 Code	接头名称 Adapter name	引物序列 Primer sequence(5′→3′)
Ea	EcoRⅠ接头 EcoRⅠadapter	CTCGTAGACTGCGTACC CATCTGACGCATGGTTAA	连接 Connection
Em	MseⅠ接头 MseⅠadapter	GACGATGAGTCCTGAG TACTCAGGACTCAT	连接 Connection
E0	EcoRⅠ+0	GACTGCGTACCAATTC	预扩增 Pre-amplification
E1	EcoRⅠ+AAC	GACTGCGTACCAATTCAAC	选择性扩增Selective amplification analysis
E2	EcoRⅠ+ACA	GACTGCGTACCAATTCACA	选择性扩增 Selective amplification analysis
E3	EcoRⅠ+ACG	GACTGCGTACCAATTCACG	选择性扩增 Selective amplification analysis
E4	EcoRⅠ+AGG	GACTGCGTACCAATTCAGG	选择性扩增 Selective amplification analysis
E5	EcoRⅠ+AAG	GACTGCGTACCAATTCAAG	选择性扩增 Selective amplification analysis
E6	EcoRⅠ+ACT	GACTGCGTACCAATTCACT	选择性扩增 Selective amplification analysis
E7	EcoRⅠ+ACC	GACTGCGTACCAATTCACC	选择性扩增 Selective amplification analysis
E8	EcoRⅠ+AGC	GACTGCGTACCAATTCAGC	选择性扩增 Selective amplification analysis
M0	MseⅠ+0	GATGAGTCCTGAGTAA	预扩增预扩增 Pre-amplification
M1	MseⅠ+CAA	GATGAGTCCTGAGTAACAA	选择性扩增 Selective amplification analysis
M2	MseⅠ+CAT	GATGAGTCCTGAGTAACAT	选择性扩增 Selective amplification analysis
M3	MseⅠ+CAC	GATGAGTCCTGAGTAACAC	选择性扩增 Selective amplification analysis
M4	MseⅠ+CAG	GATGAGTCCTGAGTAACAG	选择性扩增 Selective amplification analysis
M5	MseⅠ+ CTA	GATGAGTCCTGAGTAACTA	选择性扩增 Selective amplification analysis
M6	MseⅠ+CTT	GATGAGTCCTGAGTAACTT	选择性扩增 Selective amplification analysis
M7	MseⅠ+CTC	GATGAGTCCTGAGTAACTC	选择性扩增 Selective amplification analysis
M8	MseⅠ+CTG	GATGAGTCCTGAGTAACTG	选择性扩增 Selective amplification analysis

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 band/%
E2/M1	41	40	97.56
E2/M2	51	51	100.00
E3/M4	44	44	100.00
E3/M5	51	51	100.00
E4/M5	38	38	100.00
E4/M6	47	47	100.00
E4/M8	35	35	100.00
E5/M1	49	49	100.00
E5/M4	57	56	98.25
总计Total	413	411	99.52