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林业科学 ›› 2016, Vol. 52 ›› Issue (11): 165-169.doi: 10.11707/j.1001-7488.20161120

• 研究简报 • 上一篇    下一篇

越南黄檀锈病病原菌的鉴定

王姣, 周国英, 苏圣淞, 何苑皞, 董文统, 张茜, 刘君昂   

  1. 森林有害生物防控湖南省重点实验室 经济林培育与保护省部共建教育部重点实验室 中南林业科技大学 长沙 410004
  • 收稿日期:2016-01-08 修回日期:2016-05-16 出版日期:2016-11-25 发布日期:2016-12-16
  • 通讯作者: 刘君昂
  • 基金资助:

    林业公益性行业科研专项(201304402)。

Identification of Pathogens of the Dalbergia tonkinensis Rust Disease

Wang Jiao, Zhou Guoying, Su Shengsong, He Yuanhao, Dong Wentong, Zhang Qian, Liu Jun'ang   

  1. Hunan Provincial Key Laboratory for Control of Forest Diseases and Pests Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees of Ministry of Education Central South University of Forestry and Technology Changsha 410004
  • Received:2016-01-08 Revised:2016-05-16 Online:2016-11-25 Published:2016-12-16

摘要:

[目的] 描述在海南省澄迈县新发现的越南黄檀锈病的症状及其孢子形态特征,并提取DNA进行ITS序列扩增测序,从而鉴定该病的病原。[方法] 实地调查该病害的发生和为害情况,观察并记录病害症状。采集该病新鲜夏孢子,制备浓度为1×105个·mL-1的夏孢子悬浮液,对越南黄檀健康叶片进行涂抹接种,采用单孢子堆分离法获得1株越南黄檀锈病纯培养物HNCM1。利用孢子形态观察结合ITS序列分析的方法对该病原菌进行鉴定。[结果] 越南黄檀锈病为害寄主叶片与枝条,以当年生嫩枝叶为主,每年暴发2次,4月上旬—6月上旬与9月中旬—11月上旬为2个盛发期,重病株发病率可达75%以上。夏孢子堆初期为黄色或橙黄色,粉状,一般长在叶背面圆形或椭圆形小突起上,直径约0.2~2.5 mm,严重时连接成片引起嫩叶卷曲或畸形,后期在叶片上形成锈褐色枯死斑,导致老叶枯死或早落;夏孢子圆形、椭圆形,表面有小刺,萌发孔不明显,大小为(13~21)μm×(10~16)μm,壁厚约1 μm。冬孢子堆苍白色、胶质状,天气转凉时偶见,呈圆形,小于夏孢子堆;冬孢子椭圆形、棍棒形或近纺锤形,大小为(19~28)μm×(13~16)μm,无色,光滑单细胞,具柄,长约7~10 μm,壁厚约1 μm。该病原ITS序列扩增产物长约750 bp,产物经双向测序拼接后得到640 bp,进行Blast比对,结果表明,病原菌ITS序列与GenBank 中已有的序列同源性较低,最高仅为90%。[结论] 引起海南省澄迈县越南黄檀锈病的病原菌为紫檀无眠单胞锈菌,夏孢子ITS序列在GenBank 中尚无同源性较高序列,获取HNCM1菌株的GenBank 登录号为 KU301795。

关键词: 锈病, 越南黄檀, 紫檀无眠单胞锈菌, 海南

Abstract:

[Objective] A newly found rust disease widely infests leaves of Dalbergia tonkinensis (Papilionaceae) which is a high economic tree species for furniture in China, growing in Chengmai County, Hainan Province, but its pathogen is unknown. This study described the symptoms of the rust disease on D. tonkinensis and its spore morphology, then extracted the pathogen DNA, and amplified it by ITS, with which the pathogen was identified. [Method] The period of occurrence and the rate of disease incidence of D. tonkinensis were investigated on the spot, and symptoms of the disease were observed and recorded. In order to acquire the isolation of pathogen and purification, pathogenicity test was conducted by artificially inoculating healthy leaves of D. tonkinensis with urediniospore suspension. We obtained a strain of D. tonkinensis rust pure culture HNCM1 by single spore separation. The pathogen was identified with the method of the spore morphology observation combined with ITS sequence analysis.[Result] The rust of D. tonkinensis occurred on the leaves and twigs, mainly on young tender leaves, and outbroke 2 times a year, in early April to early June and mid September to early November, respectively. The disease incidence rate of D. tonkinensis was able to reach more than 75%. Uredinia hypophyllous was yellow, and powdery,at early stage, generally bore on the round or oval small projection of the abaxial leaf surface, 0.1-0.5 mm in diameter, scattered or gregarious, which might cause leaf roll or deformation, even death of whole leaf. Urediniospores were globoid or obovoid, (13-21) μm×(10-16) μm, yellow, echinulate, germ pores obscure. Telia were similar to uredini, pale white, colloid, found occasionally when it became cold. Teliospores were oblong-obovoid, oblong-ellipsoid or clavoid, (19-28) μm× (13-16)μm, single cell, wall ca. 1 μm thick, colourless, smooth, pedicels 7-10 μm long. ITS sequence of the pathogen was around 750 bp nucleotide long, and the product was then 640 bp after bidirectional sequencing and splicing, which only shared at most 90% homology with the recorded sequences in Gen Bank. [Conclusion] Based on morphological and molecular identification, the pathogen causing rust disease of D. tonkinensis is Maravalia pterocarpi in Chengmai County, Hainan Province, a fungal species which belongs to pucciniaceae maravalia of basidiomycota. The GenBank accession number of HNCM1 is KU301795. This is the first report of Maravalia pterocarpi paratisizing on D. tonkinensis and causing rust disease in China, as well as the first time to analyze Maravalia pterocarpi from the angle of the molecular biology.

Key words: rust disease, Dalbergia tonkinensis, Maravalia pterocarpi, Hainan

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