海南三亚和黄淮海地区玉米小斑病菌致病性及遗传多样性分析

doi:10.3864/j.issn.0578-1752.2020.06.007

Abstract

Abstract:

【Objective】The objective of this study is to clarify the pathogenicity differences, genetic diversity and relationship among the Bipolaria maydis strains from southern maize breeding base of Sanya, Hainan and Huang-Huai-Hai region, and to provide a theoretical basis for screening resistance of maize varieties under natural disease conditions in the breeding process. It also provides a reference for the popularization and utilization of maize varieties in Huang-Huai-Hai region.【Method】The pathogens were isolated from diseased leaves collected from Sanya, Hainan and Huang-Huai-Hai region, and a total of 61 strains were obtained by identification of morphology and molecular biology. The pathogenicity of 61 strains was identified by an artificial spray inoculation method, and 16 primers with clear, repetitive and polymorphic bands were used for ISSR-PCR. Popgen32 software was used to calculate the genetic distance and genetic similarity among the populations, and NTsys2.10e software was used for cluster analysis and constructing a cluster map of the pathogens in Sanya, Hainan and Huang-Huai-Hai region.【Result】The results of pathogenicity test showed that there was no weak pathogenic strain and 57.1% strong pathogenic strains isolated from Sanya, Hainan, while that of Huang-Huai-Hai region accounted for 47.5% and 17.5%, respectively. The results of genetic diversity analysis showed that the Nei’s gene diversity index (H) was 0.2820 and Shannon’s information index (I) was 0.4197 on the average level of population, indicating that there was a certain genetic variation in the pathogen. The genetic similarity coefficient was 0.9028-0.9618 and the genetic distance was 0.0390-0.1023 among different geographical populations, which indicated that the overall genetic similarity coefficient was relatively high and the genetic distance was close, but there were still different degrees of genetic differentiation among different geographical populations. Among which, the genetic similarity coefficient of strains in Henan and Hebei was the highest (GS=0.9618), and the genetic relationship was the closest; the genetic similarity coefficient of strains in Henan and Sanya was the lowest (GS=0.9028), and the genetic relationship was relatively far. The cluster analysis showed that the strains in Sanya and Huang-Huai-Hai region were obviously divided into two groups when the similarity coefficient was 0.722.【Conclusion】There are some differences in B. maydis pathogenicity between the main maize producing area of Huang-Huai-Hai region and the southern maize breeding base of Sanya, Hainan. The isolation frequency of the strong pathogenic strains in Sanya was higher than that in Huang-Huai-Hai region. The strain populations in Huang-Huai-Hai region and Sanya region have rich genetic diversity, and genetic diversity has a certain relationship with geographical origin. The genetic relationship of the strains in the adjacent regions was relatively close. However, the genetic similarity coefficient of strains between Sanya region and Huang-Huai-Hai region is relatively high, the genetic relationship is close, and also there are some gene exchanges between the two populations. Therefore, selection of resistant maize varieties or resistant parent materials under natural conditions in southern maize breeding base of Sanya has a certain effect on the yearly increase of resistance to southern corn leaf blight in Huang-Huai-Hai region, which is helpful to reduce the occurrence of disease in Huang-Huai-Hai region.

Key words: Bipolaria maydis, southern corn leaf blight, southern maize breeding base, Sanya,Hainan, Huang-Huai-Hai region, pathogenicity, genetic diversity

JiaYing CHANG,ShuSen LIU,Jie SHI,Ning GUO,HaiJian ZHANG,HongXia MA,ChunFeng YANG. Pathogenicity and Genetic Diversity of Bipolaria maydis in Sanya, Hainan and Huang-Huai-Hai Region[J].Scientia Agricultura Sinica, 2020, 53(6): 1154-1165.

Figures/Tables 5

Table 1

Pathogenicity of B. maydis in Huang-Huai-Hai region and Sanya, Hainan"

菌株编号 Strain number	采样地点 Collection site	平均发病面积Average disease area (%)	病斑大小 Lesion size (mm)	致病力 Pathogenicity	病斑描述 Lesion description
菌株编号 Strain number	采样地点 Collection site	平均发病面积Average disease area (%)	病斑大小 Lesion size (mm)	致病力 Pathogenicity	病斑颜色 Lesion color	形状 Shape	褐色边缘 Brown edge
HB1802-1	河北省保定市满城区 Mancheng, Baoding, Hebei	7.50	(2-8) × (1-2)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	-
HB1802-2	河北省保定市满城区 Mancheng, Baoding, Hebei	16.75	(1-7) × (1-1.5)	中 Moderate	深褐色 Dark brown	椭圆形 Oval	+
HB1803-1	河北省邯郸市邯郸县 Handan, Handan, Hebei	7.50	(2-7) × (1-2)	弱 Weak	黄褐色 Yellow brown	长方形Rectangle	-
HB1803-2	河北省邯郸市肥乡县 Feixiang, Handan, Hebei	27.75	(2-8) × (1-2)	中 Moderate	浅黄色 Light yellow	长方形Rectangle	+
HB1803-3	河北省邯郸市肥乡县 Feixiang, Handan, Hebei	5.00	(3-8) × (1-1.5)	弱 Weak	黄褐色 Yellow brown	长方形Rectangle	+
HB1804-1	河北省廊坊市香河县 Xianghe, Langfang, Hebei	1.75	(2-9) × (1-3)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	-
HB1804-2	河北省廊坊市固安县 Gu^’an, Langfang, Hebei	76.75	(4-7) × (1-2)	强 Strong	浅黄色 Light yellow	长方形Rectangle	-
HB1804-3	河北省廊坊市固安县 Gu^’an, Langfang, Hebei	4.25	(5-8) × (1-2)	弱 Weak	黄褐色 Yellow brown	长方形Rectangle	-
HB1804-4	河北省廊坊市香河县 Xianghe, Langfang, Hebei	8.75	(1-3) × (1-1)	弱 Weak	黄褐色 Yellow brown	长方形Rectangle	+
HB1804-5	河北省廊坊市固安县 Gu^’an, Langfang, Hebei	7.75	(4-6) × (1-1.5)	弱 Weak	浅黄色 Light yellow	椭圆形 Oval	-
HB1804-6	河北省廊坊市固安县 Gu^’an, Langfang, Hebei	10.25	(5-7) × (1-2)	中 Moderate	黄褐色 Yellow brown	长方形Rectangle	+
HB1807-1	河北省唐山市滦南县 Luannan, Tangshan, Hebei	1.25	(3-9) × (1.5-2)	弱 Weak	浅黄色 Light yellow	椭圆形 Oval	-
HB1807-2	河北省唐山市乐亭县 Laoting, Tangshan, Hebei	8.25	(2-9) × (1-2)	弱 Weak	黄褐色 Yellow brown	长方形Rectangle	+
HB1807-3	河北省唐山市滦南县 Luannan, Tangshan, Hebei	20.00	(3-8) × (1-2)	中 Moderate	深褐色 Dark brown	椭圆形 Oval	+
HB1807-4	河北省唐山市开发区 Development Zone, Tangshan, Hebei	9.50	(3-8) × (1-2)	弱 Weak	深褐色 Dark brown	长方形Rectangle	+
HB1807-5	河北省唐山市开发区 Development Zone, Tangshan, Hebei	85.50	(3-5) × (1-2)	强 Strong	浅黄色 Light yellow	长方形Rectangle	-
HB1807-6	河北省唐山市乐亭县 Laoting, Tangshan, Hebei	81.25	(1-5)×(1-2)	强 Strong	黄褐色 Yellow brown	椭圆形 Oval	+
HB1807-7	河北省唐山市乐亭县 Laoting, Tangshan, Hebei	72.00	(3-7) × (1-2)	强 Strong	黄褐色 Yellow brown	椭圆形 Oval	+
HB1807-8	河北省唐山市迁安 Qian^’an, Tangshan, Hebei	13.25	(2-6) × (1-2)	中 Moderate	浅黄色 Light yellow	椭圆形 Oval	-
HB1807-9	河北省唐山市迁安 Qian^’an, Tangshan, Hebei	24.50	(1-4) × (1-2)	中 Moderate	黄褐色 Yellow brown	椭圆形 Oval	+
HB1807-10	河北省唐山市玉田县 Yutian, Tangshan, Hebei	9.50	(2-8) × (1-1.5)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	-
HB1807-11	河北省唐山市开发区 Development Zone, Tangshan, Hebei	58.00	(2-6) × (1-2)	强 Strong	浅黄色 Light yellow	椭圆形 Oval	+
HB1807-12	河北省唐山市滦南县 Luannan, Tangshan, Hebei	2.75	(4-8) × (1-2)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	-
HB1808-1	河北省衡水市深州市 Shenzhou, Hengshui, Hebei	27.50	(3-9) × (1-1.5)	中 Moderate	深褐色 Dark brown	长方形Rectangle	+
HB1808-2	河北省衡水市深州市 Shenzhou, Hengshui, Hebei	13.50	(2-9) × (1-2)	中 Moderate	深褐色 Dark brown	长方形Rectangle	+
HB1808-3	河北省衡水市深州市 Shenzhou, Hengshui, Hebei	17.50	(3-8) × (1-2)	中 Moderate	浅黄色 Light yellow	长方形Rectangle	-
HB1808-4	河北省衡水市深州市 Shenzhou, Hengshui, Hebei	9.50	(2-5) × (1-2)	弱 Weak	浅黄色 Light yellow	椭圆形 Oval	-
HN1802-1	河南省鹤壁市浚县 Xunxian, Hebi, Henan	12.50	(4-9) × (1-2)	中 Moderate	深褐色 Dark brown	长方形Rectangle	-
HN1802-2	河南省鹤壁市浚县 Xunxian, Hebi, Henan	3.25	(1-8) × (1-1.5)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	-
HN1802-3	河南省鹤壁市浚县 Xunxian, Hebi, Henan	12.50	(4-6) × (1-1.5)	中 Moderate	深褐色 Dark brown	椭圆形 Oval	-
HN1802-4	河南省鹤壁市浚县 Xunxian, Hebi, Henan	9.50	(4-7) × (1-1.5)	弱 Weak	深褐色 Dark brown	长方形Rectangle	-
HN1803-1	河南省安阳市滑县 Huaxian, Anyang, Henan	13.25	(1-7) × (1-1.5)	中 Moderate	深褐色 Dark brown	长方形Rectangle	+
HN1804-1	河南省新乡市辉县 Huixian, Xinxiang, Henan	15.00	(5-8) × (1-1.5)	中 Moderate	深褐色 Dark brown	长方形Rectangle	+
HN1807-1	河南省漯河市召陵区 Zhaoling, Luohe, Henan	77.50	(1-8) × (1-2)	强 Strong	黄褐色 Yellow brown	长方形Rectangle	+
HN1807-2	河南省漯河市源汇区 Yuanhui, Luohe, Henan	3.25	(3-7) × (1-2)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	-
HN1807-3	河南省漯河市临颍县 Linying, Luohe, Henan	4.00	(3-8) × (1-2)	弱 Weak	深褐色 Dark brown	长方形Rectangle	+
HN1810-1	河南省周口市西华县 Xihua, Zhoukou, Henan	13.50	(4-6) × (1-1.5)	中 Moderate	深褐色 Dark brown	椭圆形 Oval	+
SD1811-1	山东青岛市平度市 Pingdu, Qingdao, Shandong	6.50	(3-8) × (1-1.5)	弱 Weak	浅黄色 Light yellow	长方形Rectangle	+
SD1811-2	山东省青岛市平度市 Pingdu, Qingdao, Shandong	3.50	(2-8) × (1-1.5)	弱 Weak	黄褐色 Yellow brown	长方形Rectangle	+
SD1812-1	山东省东营市利津县 Lijin, Dongying, Shandong	36.25	(2-9) × (1-2)	强 Strong	深褐色 Dark brown	椭圆形 Oval	-
SY-1	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	20.00	(3-7) × (1-1.5)	中 Moderate	深褐色 Dark brown	长方形Rectangle	+
SY-2	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	18.75	(1-8) × (1-2)	中 Moderate	黄褐色 Yellow brown	长方形Rectangle	+
SY-3	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	21.25	(1-9) × (1-2)	中 Moderate	黄褐色 Yellow brown	椭圆形 Oval	-
SY-4	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	18.25	(1-7) × (0.5-2)	中 Moderate	浅黄色 Light yellow	长方形Rectangle	-
SY-5	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	70.25	(2-8) × (1-1.5)	强 Strong	黄褐色 Yellow brown	长方形Rectangle	-
SY-6	海南省三亚市吉阳区 Jiyang, Sanya, Hainan	57.00	(2-8) × (1-1.5)	强 Strong	深褐色 Dark brown	长方形Rectangle	-
SY-7	海南省三亚市吉阳区 Jiyang, Sanya, Hainan	52.00	(2-8) × (1-2)	强 Strong	深褐色 Dark brown	长方形Rectangle	+
SY-8	海南省三亚市吉阳区 Jiyang, Sanya, Hainan	25.00	(4-7) × (1-2)	中 Moderate	黄褐色 Yellow brown	长方形Rectangle	-
SY-9	海南省三亚市吉阳区 Jiyang, Sanya, Hainan	10.25	(1-8) × (1-2)	中 Moderate	黄褐色 Yellow brown	椭圆形 Oval	-
SY-10	海南省三亚市天涯区 Tianya, Sanya, Hainan	33.75	(1-7) × (0.5-2)	强 Strong	黄褐色 Yellow brown	长方形Rectangle	-
SY-11	海南省三亚市天涯区 Tianya, Sanya, Hainan	52.50	(1-9)×(1-1.5)	强 Strong	浅黄色 Light yellow	长方形Rectangle	-
SY-12	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	54.50	(2-8) × (1-1.5)	强 Strong	浅黄色 Light yellow	长方形Rectangle	-
SY-13	海南省三亚市海棠区 Haitang, Sanya, Hainan	24.50	(2-9) × (1-1.5)	中 Moderate	浅黄色 Light yellow	椭圆形 Oval	-
SY-14	海南省三亚市海棠区 Haitang, Sanya, Hainan	67.50	(5-8) × (1-2)	强 Strong	深褐色 Dark brown	长方形Rectangle	+
SY-15	海南省三亚市海棠区 Haitang, Sanya, Hainan	23.25	(5-8) × (1-1.5)	中 Moderate	黄褐色Yellow brown	椭圆形 Oval	+
SY-16	海南省三亚市海棠区 Haitang, Sanya, Hainan	86.25	(1-7) × (0.5-1.5)	强 Strong	浅黄色 Light yellow	椭圆形 Oval	-
SY-17	海南省三亚市海棠区 Haitang, Sanya, Hainan	92.50	(1-8) × (1-2)	强 Strong	浅黄色 Light yellow	长方形Rectangle	-
SY-18	海南省乐东黎族自治县九所镇 Jiusuo, Ledong Li Autonomous County, Hainan	64.00	(4-8) × (1-2)	强 Strong	浅黄色 Light yellow	长方形Rectangle	-
SY-19	海南省三亚市陵水县 Lingshui, Sanya, Hainan	12.25	(3-8) × (1-1.5)	中 Moderate	浅黄色 Light yellow	长方形Rectangle	-
SY-20	海南省三亚市陵水县 Lingshui, Sanya, Hainan	38.75	(2-8) × (1-1.5)	强 Strong	黄褐色 Yellow brown	长方形Rectangle	+
SY-21	海南省三亚市陵水县 Lingshui, Sanya, Hainan	38.25	(3-8) × (1-1.5)	强 Strong	深褐色 Dark brown	椭圆形Oval	+

Table 1

Table 2

Table 3

Table 4

Fig. 1

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引物 Primer	序列 Sequence (5′-3′)	退火温度 Tm (℃)	扩增条带数 Number of amplified bands	多态性条带 Number of polymorphic bands	多态性比率 Polymorphism percent (%)
807	(AG)₈T	46	10	8	80.00
808	(AG)₈C	46	17	14	82.35
812	(GA)₈A	45	7	7	100.00
825	(AC)₈T	50	8	5	62.50
834	(AG)₈YT	48	14	14	100.00
835	(AG)₈YC	50	12	11	91.67
841	(GA)₈YC	48	16	14	87.50
856	(AC)₈YA	50	8	7	87.50
857	(AC)₈YG	51	12	8	66.67
880	(GGAGA)₃	47	12	11	91.67
884	HBHA(GA)₆G	46	10	8	80.00
885	BHB(GA)₇	46	11	8	72.73
887	DVD(TC)₇	45	12	6	50.00
888	BDB(CA)₇	49	9	4	44.44
889	DBD(AC)₇	49	9	8	88.89
891	HVH(TG)₇	50	10	7	70.00
总计 Total			177	140	79.10

	多态性位点 Number of polymorphic loci	多态性比率 Polymorphism percent (%)	观察等位基因数 Observed number of alleles (Na)	有效等位基因 Effective number of alleles (Ne)	基因多样性指数Nei’s gene diversity index (H)	Shannon’s信息指数 Shannon’s information index (I)
河北HB	127	71.75	1.7175	1.4930	0.2783	0.4072
河南HN	100	56.50	1.5650	1.3822	0.2137	0.3134
山东SD	75	42.37	1.4237	1.3060	0.1717	0.2505
三亚SY	121	68.36	1.6836	1.4186	0.2437	0.3637
群体水平 Population level	140	79.10	1.7910	1.4904	0.2820	0.4197

种群 Group	河北HB	山东SD	河南HN	三亚SY
河北HB	****	0.9455	0.9618	0.9404
山东SD	0.0560	****	0.9351	0.9116
河南HN	0.0390	0.0671	****	0.9028
三亚SY	0.0614	0.0925	0.1023	****

Pathogenicity and Genetic Diversity of Bipolaria maydis in Sanya, Hainan and Huang-Huai-Hai Region

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