Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (5): 822-836.doi: 10.3864/j.issn.0578-1752.2019.05.005

• PLANT PROTECTION • Previous Articles     Next Articles

Genetic Diversity Analysis of Curvularia lunata in Summer Maize in Huang-Huai-Hai Region

CHANG JiaYing,LIU ShuSen,MA HongXia,SHI Jie(),GUO Ning,ZHANG HaiJian   

  1. Plant Protection Institute of Hebei Academy of Agricultural and Forestry Sciences/Key Laboratory of IPM on Crops in Northern Region of North China, Ministry of Agriculture/IPM Centre of Hebei Province, Baoding 071000, Hebei
  • Received:2018-10-27 Accepted:2018-12-23 Online:2019-03-01 Published:2019-03-12
  • Contact: Jie SHI E-mail:shij99@163.com

Abstract:

【Objective】The objective of this study is to identify the genetic diversity and relationship of Curvularia lunata , the causal agent of maize Curvularia leaf spot occurred in different regions and years in Huang-Huai-Hai region via molecular techniques, and to provide scientific data for studying the occurrence and prevalence of the disease. 【Method】 The samples collected from five provinces (Henan, Hebei, Shandong, Anhui and Jiangsu) in Huang-Huai-Hai region in 2013, 2016 and 2017 were separated and identified by morphology and molecular biology (ITS and EF-1α sequence analysis). A total of 175 C. lunata strains were obtained. Thirteen highly polymorphic and reproducible ISSR primers were screened out from universal primers developed by Columbia University and were used to amplify the 175 C. lunata strains. Popgen32 software was used to calculate the polymorphism ratio, Shannon’s information index, genetic distance and genetic similarity among populations. UPGMA cluster analysis and principal coordinate analysis based on genetic similarity coefficient were carried out by NTsys 2.10e software, and cluster analysis map and scatter plot were constructed. 【Result】 A total of 105 polymorphic bands were obtained with a polymorphism ratio of 100%. At the group average level, the level of genetic diversity (H) was 0.3867, and Shannon’s information index (I) was 0.5682, indicating that the C. lunata had abundant genetic diversity. The genetic diversity among different geographical populations was different. The genetic diversity of Henan and Anhui populations was the highest, and that of Jiangsu population was low. The flora of the same geographical origin was far away from each other during the year, and the flora from different geographical origins in the same year was relatively close. Cluster analysis showed that the similarity coefficients of all strains were 0.51-0.93. At the similarity coefficient of 0.59 level, 175 strains were divided into 2 groups and 5 subgroups. There were annual differences among the subgroups, gene exchanges among the different geographical populations, and the genetic correlation was high. The results of principal component analysis were consistent with those of cluster analysis, and the strains in the same year were obviously clustered together.【Conclusion】There was a high genetic variation in the pathogen populations causing maize Curvularia leaf spot disease in Huang-Huai-Hai region, and the genetic relationship of the adjacent pathogens was close. The strains from the same area showed a certain genetic distance in different years, while the genetic distance of strains of different geographical origins in the same year was close. The results of the analysis indicated that the main pathogen causing the leaf spot disease of C. lunata was not mainly from local source, they probably came from the southern rice and turf grass or maize production areas of Southeast Asia, but occasionally a few strains that survived on the ground surface could be the initial source of infection in the following year.

Key words: Curvularia lunata, Huang-Huai-Hai region, genetic diversity, ISSR

Table 1

The tested strains of C. lunata "

采集年份
Year
采集地点
Collection site
菌株编号
Strain number
2013 安徽
Anhui
宿州Suzhou 萧县马井镇Majing, Xiaoxian AH1301-1, AH1301-2
灵璧县灵城镇Lingcheng, Lingbi AH1301-6—AH1301-9
宿县黄口镇Huangkou, Suxian AH1301-10
淮北Huaibei 濉溪县孙疃镇Suntuan, Suixi AH1301-3—AH1301-5
亳州Bozhou 蒙城县许疃镇Xutuan, Mengcheng AH1302-1—AH1302-3
蒙城县东土镇Dongtu, Mengcheng AH1302-4
蒙城县板桥集镇Banqiaoji, Mengcheng AH1302-5, AH1302-6
蒙城县王集乡Wangji, Mengcheng AH1302-7—AH1302-9
利辛县巩店镇Gongdian, Lixin AH1302-10, AH1302-11
利辛县汝集镇Ruji, Lixin AH1302-12—AH1302-15
河南
He’nan
驻马店Zhumadian 确山县朱古洞乡Zhugudong, Queshan HN1301-1—HN1301-4
确山县三里河乡Sanlihe, Queshan HN1301-5
泌阳县盘古乡Pangu, Miyang HN1301-6—HN1301-9
泌阳县高店乡Gaodian, Miyang HN1301-10
遂平县车站镇Chezhan, Suiping HN1301-11—HN1301-16
遂平县和兴乡Hexing, Suiping HN1301-17—HN1301-19
周口Zhoukou 商水县姚集乡Yaoji, Shangshui HN1302-1
商水县城关镇Chengguan, Shangshui HN1302-2
沈丘县白集镇Baiji, Shenqiu HN1302-3—HN1302-5
沈丘县北校乡Beixiao, Shenqiu HN1302-6—HN1302-8
平顶山Pingdingshan 叶县龚店乡Gongdian, Yexian HN1303-1, HN1303-2
叶县叶邑镇Yeyi, Yexian HN1303-3, HN1303-4
叶县田庄乡Tianzhuang, Yexian HN1303-5, HN1303-6
许昌Xuchang 许昌县尚积镇Shangji, Xuchang HN1304-1, HN1304-2
许昌县蒋李集镇Jiangliji, Xuchang HN1304-3, HN1304-4
开封Kaifeng 兰考县城关镇Chengguan, Lankao HN1305-1, HN1305-2
商丘Shangqiu 拓城县胡囊镇Hu’nang, Tuocheng HN1306-1, HN1306-2
2016 安徽Anhui 宿州Suzhou 埇桥区Yongqiao AH1601-1
河南
He’nan
商丘Shangqiu 夏邑县曹集乡Caoji, Xiayi HN1606-1
周口Zhoukou 太康县张集镇Zhangji, Taikang HN1602-1, H1N602-2
漯河Luohe 舞阳县孟寨镇Mengzhai, Wuyang HN1607-1, HN1607-2
开封Kaifeng 杞县李庄村Lizhuang, Qixian HN1605-1, HN1605-2
兰考县城关乡Chengguan, Lankao HN1605-3
郑州Zhengzhou 荥阳县高村乡Gaocun, Xingyang HN1608-1, HN1608-2
巩义市河洛镇Heluo, Gongyi HN1608-3, HN1608-4
焦作Jiaozuo 沁阳市崇义镇Chongyi, Qinyang HN1609-1
博爱县清化镇Qinghua, Boai HN1609-2, HN1609-3
修武县郇封镇千仓村Qiancang, Xunfeng, Xiuwu HN1609-4, HN1609-5
武陟县宁郭镇张庄村Zhangzhuang, Ningguo, Wuzhi HN1609-6
采集年份
Year
采集地点
Collection site
菌株编号
Strain number
2017 安徽
Anhui
亳州Bozhou 利辛县垄店镇Gongdian, Lixin AH1702-1
蒙城县王集乡Wangji, Mengcheng AH1702-2
蒙城县板桥集镇Banqiaoji, Mengcheng AH1702-3
宿州Suzhou 灵璧县灵城镇Lingcheng, Lingbi AH1701-1, AH1701-2
埇桥区大店镇Dadian, Yongqiao AH1701-3
埇桥区Yongqiao AH1701-4
淮北Huaibei 濉溪县刘桥镇Liuqiao, Suixi AH1703-2
相山区Xiangshan AH1703-3
濉溪县南平镇Nanping, Suixi AH1703-1
河北
Hebei
石家庄Shijiazhuang 新乐市马头铺镇Matoupu, Xinle HB1701-1
赵县赵州镇Zhaozhou, Zhaoxian HB1701-2
藁城区Gaocheng HB1701-3, HB1701-5, HB1701-6
灵寿县北洼乡Beiwa, Lingshou HB1701-4
保定Baoding 望都韩庄镇Hanzhuang,Wangdu HB1702-1
蠡县南庄镇Nanzhuang, Lixian HB1702-2
雄县双堂乡Shuangtang, Xiongxian HB1702-3
蠡县蠡吴镇Liwu, Lixian HB1702-4
高阳县高阳镇Gaoyang, Gaoyang HB1702-5
满城大册营镇Daceying, Mancheng HB1702-6
易县高村镇Gaocun, Yixian HB1702-7
涞源王安镇Wang’an, Laiyuan HB1702-8
易县尉都乡Weidu, Yixian HB1702-9
涞源县胡家庄乡Hujiazhuang, Laiyuan HB1702-10
涿州市松林店镇Songlindian, Zhuozhou HB1702-11
定兴县定兴镇Dingxing, Dingxing HB1702-12
徐水区Xushui HB1702-13
邯郸Handan 邯郸县丛台区Congtai, Handan HB1703-1
峰峰县新坡镇Xinpo, Fengfeng HB1703-2
大城县大尚屯镇Dashangtun, Dacheng HB1706-3
大城县广安镇Guang’an, Dacheng HB1706-4
衡水Hengshui 武邑县肖桥头镇Xiaoqiaotou, Wuyi HB1704-3, HB1704-4
故城县故城镇Gucheng, Gucheng HB1704-1
深州市Shenzhou HB1704-2
沧州Cangzhou 东光县找王镇Zhaowang, Dongguang HB1705-1
青县新兴镇Xinxing, Qingxian HB1705-2
沧县崔尔庄镇Cuierzhuang, Cangxian HB1705-3
河间市时村乡Shicun, Hejian HB1705-4
任丘市麻家坞镇Majiawu, Renqiu HB1705-5
青县曹寺镇Caosi, Qingxian HB1705-6
献县淮镇Huaizhen, Xianxian HB1705-7
廊坊Langfang 固安县马庄镇Mazhuang, Gu’an HB1706-1
固安县Gu’an HB1706-2
固安县牛驼镇Niutuo, Gu’an HB1706-5
采集年份
Year
采集地点
Collection site
菌株编号
Strain number
唐山Tangshan 玉田县杨家套乡Yangjiatao, Yutian HB1707-1
昌黎县葛条港乡Getiaogang, Changli HB1707-2
玉田县石臼窝镇Shijiuwo, Yutian HB1707-3
滦州市滦州镇Luanzhou, Luanzhou HB1707-6
滦州市Luanzhou HB1707-7
遵化市团瓢庄乡Tuanpiaozhuang, Zunhua HB1707-8
迁安市Qian’an 五重安乡Wuchongan HB1707-4
赵新庄Zhaoxinzhuang HB1707-5
承德Chengde 承德县下板城镇Xiabancheng, Chengde HB1709-1
秦皇岛Qinhuangdao 卢龙县Lulong HB1708-1
抚宁区Funing HB1708-2
河南
He’nan
安阳Anyang 安阳县崔家桥镇Cuijiaqiao, Anyang HN1710-1
鹤壁Hebi 淇县西岗镇Xigang, Qixian HN1711-1
新乡Xinxiang 辉县胡桥乡Huqiao, Huixian HN1712-1
新乡县Xinxiang HN1712-2
漯河Luohe 源汇区Yuanhui HN1707-1
驻马店Zhumadian 西平县二郎镇Erlang, Xiping HN1701-1
周口Zhoukou 西华县田口乡Tiankou, Xihua HN1702-1
平顶山Pingdingshan 宝丰县杨庄镇Yangzhuang, Baofeng HN1703-1
商丘Shangqiu 柘城县Zhecheng HN1706-1
江苏
Jiangsu
徐州Xuzhou 铜山区大彭镇Dapeng, Tongshan JS1701-1, JS1701-2
沛县栖山镇Qishan, Peixian JS1701-3
沛县鹿楼镇Lulou, Peixian JS1701-4
沛县朱寨镇Zhuzhai, Peixian JS1701-5
山东
Shandong
滨州Binzhou 阳信县劳店镇Laodian, Yangxin SD1701-1
德州Dezhou 乐陵县Leling SD1702-1
平原县Pingyuan SD1702-2
德城区Decheng SD1702-3
济宁Jining 微山县Weishan SD1704-1
曲阜市石门山镇Shimenshan, Qufu SD1704-2
汶上县苑庄镇Yuanzhuang, Wenshang SD1704-3
嘉祥县瞳里镇Tongli, Jiaxiang SD1704-4
任城区Rencheng SD1704-5
枣庄Zaozhuang 滕州市西岗镇Xigang,Tengzhou SD1703-1
峄城区峨山镇Eshan, Yicheng SD1703-2
临沂Linyi 兰陵县金陵镇Jinling, Lanling SD1705-1
兰山区汪沟镇Wanggou, Lanshan SD1705-2
莱芜Laiwu 莱芜区杨庄镇Yangzhuang, Laiwu SD1706-1
泰安Taian 东平县彭集镇Pengji, Dongping SD1707-1
菏泽Heze 鄄城县红船镇Hongchuan, Juancheng SD1708-1
聊城Liaocheng 聊城市东昌府区Dongchangfu, Liaocheng SD1709-1
潍坊Weifang 临朐县Linqu SD1710-1

Table 2

The amplification result with used primers"

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 ratio (%)
808 (AG)8C 49 10 10 100
810 (GA)8T 45 7 7 100
823 (TC)8C 50 6 6 100
827 (AC)8G 52 7 7 100
835 (AG)8YC 53 10 10 100
841 (GA)8YC 50 11 11 100
842 (GA)8YG 50 10 10 100
847 (CA)8RC 54 6 6 100
855 (AC)8YT 46 7 7 100
873 (GACA)4 49 7 7 100
880 (GGAGA)3 46 7 7 100
888 BDB(CA)7 48 9 9 100
891 HVH(TG)7 47 8 8 100
总计Total 105 105 100

Fig. 1

Amplification results of genome DNA of some C. lunata strains with four primers"

Table 3

Genetic diversity parameters among geographic populations of C. lunata "

多态性位点
Number of polymorphic loci
多态性比率
Polymorphism
ratio (%)
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 (H) Shannon’s信息指数
Shannon’s information index (I)
安徽AH 103 98.10 1.9810 1.6344 0.3665 0.5428
河南HN 105 100.00 2.0000 1.6309 0.3662 0.5438
河北HB 96 91.43 1.9143 1.5855 0.3415 0.5072
江苏JS 58 55.24 1.5524 1.3904 0.2170 0.3171
山东SD 90 85.71 1.8571 1.5355 0.3109 0.4623
群体水平
Population level
105 100.00 2.0000 1.6761 0.3867 0.5682

Table 4

Genetic identity and genetic distance among geographic populations of C. lunata "

种群Group 安徽AH13 河南HN13 安徽AH16 河南HN16 安徽AH17 河北HB17 河南HN17 江苏JS17 山东SD17
安徽AH13 0.9869 0.6550 0.8571 0.7644 0.8767 0.8621 0.7855 0.8652
河南HN13 0.0132 0.6467 0.8559 0.7682 0.8829 0.8623 0.7854 0.8669
安徽AH16 0.4232 0.4358 0.7510 0.6771 0.6893 0.6751 0.6316 0.6953
河南HN16 0.1542 0.1557 0.2863 0.7542 0.8292 0.7847 0.7424 0.8173
安徽AH17 0.2687 0.2637 0.3899 0.2821 0.8698 0.9014 0.9435 0.8947
河北HB17 0.1316 0.1246 0.3720 0.1873 0.1395 0.8924 0.8809 0.9491
河南HN17 0.1483 0.1481 0.3929 0.2424 0.1038 0.1139 0.8897 0.9221
江苏JS17 0.2415 0.2415 0.4594 0.2978 0.0582 0.1269 0.1168 0.9014
山东SD17 0.1448 0.1429 0.3635 0.2018 0.1112 0.0522 0.0811 0.1038

Fig. 2

Dendrogram obtained by UPGMA clustering for C. lunata "

Fig. 3

Three-dimensional scatter plot of principal coordinate analysis of C. lunata Number of C. lunata strains;Coordinate axis 1, 2, 3 represent the contribution rate of the first three principal components to genetic variation, respectively"

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