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作物学报 ›› 2022, Vol. 48 ›› Issue (3): 770-777.doi: 10.3724/SP.J.1006.2022.14035

• 研究简报 • 上一篇    

中国黄麻炭疽病病原菌的分离鉴定及系统发育分析

郭艳春1,2,3**(), 姚嘉瑜1,2,3**(), 张镕斌1,2, 陈思远1,2,3, 何青垚1,2,3, 陶爱芬1,2, 方平平1,2, 祁建民1,2, 张列梅1, 张立武1,2,3,*()   

  1. 1福建农林大学农学院/作物遗传育种与综合利用教育部重点实验室/福建省作物设计育种重点实验室, 福建福州 350002
    2福建农林大学农业农村部东南黄红麻实验观测站/福建省麻类种质资源共享平台/福建省南方经济作物遗传育种与多用途开发国际科技合作基地, 福建福州 350002
    3福建农林大学海峡联合研究院基因组与生物技术中心, 福建福州 350002
  • 收稿日期:2021-02-26 接受日期:2021-07-12 出版日期:2022-03-12 网络出版日期:2021-08-05
  • 通讯作者: 张立武
  • 作者简介:郭艳春, E-mail: 565358452@qq.com
    姚嘉瑜, E-mail: 3533551417@qq.com第一联系人:**同等贡献
  • 基金资助:
    国家自然科学基金项目(31771369);国家现代农业产业技术体系建设专项(CARS-19-E06)

Identification and phylogenetic analysis of the pathogen of jute anthracnose in China

GUO Yan-Chun1,2,3**(), YAO Jia-Yu1,2,3**(), ZHANG Rong-Bin1,2, CHEN Si-Yuan1,2,3, HE Qing-Yao1,2,3, TAO Ai-Fen1,2, FANG Ping-Ping1,2, QI Jian-Min1,2, ZHANG Lie-Mei1, ZHANG Li-Wu1,2,3,*()   

  1. 1Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops/Fujian Key Laboratory for Crop Breeding by Design/College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
    2Experiment Station of Ministry of Agriculture and Rural Affairs for Jute and Kenaf in Southeast China/Public Platform of Fujian for Germplasm Resources of Bast Fiber Crops/Fujian International Science and Technology Cooperation Base for Genetics, Breeding and Multiple Utilization Development of Southern Economic Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
    3Center for Genomics and Biotechnology, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China.
  • Received:2021-02-26 Accepted:2021-07-12 Published:2022-03-12 Published online:2021-08-05
  • Contact: ZHANG Li-Wu
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    National Natural Science Foundation of China(31771369);China Agriculture Research System(CARS-19-E06)

摘要:

炭疽病是严重影响黄麻纤维产量和品质的主要真菌性病害, 明确炭疽病病原菌种类并测定其致病力有助于黄麻抗病育种和优异抗病基因资源挖掘。本研究从福建省福州市、福建省漳州市、河南省信阳市、安徽省六安市、浙江省杭州市、广西省南宁市和湖南省长沙市等7个黄麻生产区采集黄麻炭疽病病原菌样本, 分离纯化出92个菌株; rDNA-ITS区域的序列分析表明, 其中11个为典型炭疽病病原菌。LSU区域的系统进化树和形态学特征鉴定显示, ZZ4、GX19等10个菌株为胶胞炭疽菌(Colletotrichum gloeosporioides), 菌株CS3为黑线炭疽菌(Colletotrichum dematium)。人工接种的致病力测定表明, 不同炭疽菌菌株致病力存在显著差异, 其中胶胞炭疽菌菌株GX19致病力最强, 表现为优势菌株。这些结果为黄麻炭疽病抗性基因位点挖掘和有效防治奠定了基础。

关键词: 黄麻, 炭疽菌, 致病力测定, 系统发育

Abstract:

Anthracnose is a major fungal disease that seriously affects the yield and quality in jute fiber. Identifying the types of anthracnose pathogens and determining their pathogenicity will help jute disease-resistant breeding and develop elite disease-resistant gene resources. The 92 strains were isolated and purified from samples of diseased jute, which were collected from 7 jute production areas including Fuzhou and Zhangzhou in Fujian Province, Xinyang City in Henan Province, Liu’an City in Anhui Province, Hangzhou City in Zhejiang Province, Nanning City in Guangxi Province, and Changsha City in Hunan Province. The sequence analysis of the rDNA-ITS region showed that 11 out of the 92 strains were typical anthracnose pathogens. Further identification of the phylogenetic tree of the LSU region and morphological characteristics revealed that 10 strains, including ZZ4 and GX19, were Colletotrichum gloeosporioides, while the strain of CS3 was Colletotrichum dematium. The pathogenicity test of artificial inoculation indicated that there was a significant difference in pathogenicity between Colletotrichum gloeosporioides and Colletotrichum dematium. These results laid a basis for identification of resistance gene loci and effective controlling of anthracnose in jute.

Key words: jute, Colletotrichum, pathogenicity determination, phylogenetic tree

表1

本研究rDNA-ITS和LSU的PCR扩增引物序列"

扩增区域名称
Amplified region name
引物名称
Primer name
引物序列
Primer sequence (5°-3°)
退火温度
Annealing temperature (℃)
rDNA-ITS ITS1 TCCGTAGGTGAACCTGCGG 55
rDNA-ITS ITS4 TCCTCCGCTTATTGATATGC
LSU NL1 GCATATCAATAAGCGGAGGAAAAG 57
LSU NL4 GGTCCGTGTTTCAAGACGG

图1

黄麻不同组织田间发病图片 a: 茎秆染病症状; b: 蒴果染病症状; c: 叶片染病症状。"

图2

黄麻92个病原菌菌株rDNA-ITS序列系统发育树"

图3

黄麻11个炭疽病病原菌的LSU序列系统发育树"

图4

黄麻11个典型炭疽病菌株的菌落、分生孢子和附着孢形态 a1、b1、c1、d1、e1、f1、g1、h1、i1、j1、k1分别是ZZ4、ZZ5、ZZ6、XY13、GX9、GX10、GX17、GX19、CS5、CS13、CS3炭疽菌的菌落正面; a2、b2、c2、d2、e2、f2、g2、h2、i2、j2、k2分别是ZZ4、ZZ5、ZZ6、XY13、GX9、GX10、GX17、GX19、CS5、CS13、CS3炭疽菌的菌落反面; a3、b3、c3、d3、e3、f3、g3、h3、i3、j3、k3分别是ZZ4、ZZ5、ZZ6、XY13、GX9、GX10、GX17、GX19、CS5、CS13、CS3炭疽菌的分生孢子; a4、b4、c4、d4、e4、f4、g4、h4、i4、j4、k4分别是ZZ4、ZZ5、ZZ6、XY13、GX9、GX10、GX17、GX19、CS5、CS13、CS3炭疽菌的附着孢子。"

表2

福黄麻3号和福农5号对11个炭疽菌菌株的抗性表现"

菌株名称
Strain
福黄麻3号病斑平均大小
Average size of Fuhuangma 3 lesions
菌株名称
Strain
福农5号病斑平均大小
Average size of Funong 5 lesions
福州地区Fuzhou region
GX19 11.70 a GX19 16.40 a
XY13 10.30 ab CS13 13.60 ab
CS13 9.80 ab CS5 13.40 abc
CS5 9.70 ab ZZ6 12.80 abc
CS3 9.50 ab ZZ5 11.90 abcd
GX17 9.10 ab GX17 11.70 abcde
GX9 9.00 ab XY13 11.60 abcde
ZZ5 9.00 ab GX10 10.90 bcde
GX10 7.90 ab CS3 8.40 cde
ZZ6 7.80 ab GX9 7.20 de
ZZ4 5.20 b ZZ4 6.80 e
三明地区Sanming region
GX19 13.00 a GX19 13.40 a
GX9 11.10 ab GX9 12.90 a
ZZ5 10.60 bc ZZ5 12.40 a
CS3 10.10 bcd ZZ4 11.50 ab
XY13 9.30 bcde XY13 11.40 ab
GX17 9.20 bcde CS13 10.80 ab
ZZ4 8.70 cde GX17 10.70 ab
ZZ6 8.60 cde ZZ6 10.70 ab
CS13 8.10 def CS3 9.10 bc
CS5 7.30 ef CS5 7.00 c
GX10 6.40 f GX10 6.90 c
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