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中国首次报道禾谷根结线虫侵染香蕉

First Report of Meloidogyne graminicola Infecting Banana in China.

作者信息

Zhou X, Liu G K, Xiao S, Zhang S S

机构信息

Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, 350002, Fuzhou, Fujian, P. R. China.

出版信息

Plant Dis. 2015 Mar;99(3):420. doi: 10.1094/PDIS-08-14-0810-PDN.

Abstract

Bananas (Musa spp.) are one of world's most popular fruits, and China is the third largest banana-producing country in the world. Root-knot nematodes, Meloidogyne spp., are common pests of banana worldwide, but damage to this crop caused by M. graminicola has not been reported up to now. During a survey of root-knot nematode species infecting banana in Fujian Province, China, swollen, galled primary and secondary root samples of Musa nana cv. Tianbao (AAA) were collected from two commercial fields in Nanjing County in May 2013. The affected plants did not exhibit obvious above-ground symptoms. Seriously infected roots were malformed and dehiscent, with the tissue discolored and rotting. Examination of symptomatic roots revealed one to several females of Meloidogyne sp. within each gall, with egg masses that were often completely embedded within the gall without protruding through the root surface, and with second-stage juveniles (J2) hatched inside the galls. Population densities of this nematode ranged from 452 to 2,056 eggs and J2 per 5 g of fresh roots. Males were rarely observed. Morphological measurements of 25 females and 20 J2 matched the original description of M. graminicola (1). The perineal patterns of females were dorsoventrally ovoid, with low to moderately high and round dorsal arches and lacking obvious lateral lines; striae were smooth and some were broken by a few obvious irregular, zig-zag striae in the dorsal part of the pattern; phasmids were close together (13.1 to 19.7 μm). The J2 had long tapered tails (63.4 to 75.5 μm), with long narrow hyalines (13.1 to 19.9 μm) and marked clavate termini. DNA was extracted from one mature female. The ITS1-5.8S-ITS2 rDNA region was amplified with V5367/26S (TTGATTACGTCCCTGCCCTTT/TTTCACTCGCCGTTACTAAGG) (2) and the COII and IRNA mtDNA genes were amplified with C2F3/MRH106 (GGTCAATG TTCAGAAATTTGTGG/AATTTCTAAAGACTTTTCTTAGT) (3) and then sequenced. The sequences were subjected to a database search using BLAST to verify the identity. Sequences from the ITS region were 788 bp (GenBank Accession Nos. KM111531 and KM236560) and were 96.8 to 99.1% identical to the known sequences of M. graminicola in Genbank. Sequences from the mtDNA were 666 bp (KM111533 and KM236559) and showed 99.1 to 99.4% homology with the known sequences of M. graminicola (KJ139963 and HG529223). In glasshouse tests, banana plantlets (M. nana cv. Tianbao) about 20 cm high were transplanted in 25-cm-diameter pots and inoculated with 5,000 J2 of each collected population of M. graminicola replicated six times; a noninoculated control was included. After 15 weeks, all inoculated plants were stunted and chlorotic. Galling symptoms on roots were similar to those in the field, and dissection of galled root tissue revealed that different life stages of the nematode were present, with population densities ranging from 1,238 to 6,562 eggs and J2 per 5 g of fresh roots. The noninoculated control plants grew well and had no galling symptoms on the roots. These results confirmed the nematodes' pathogenicity on banana. On the basis of these results, the root-knot nematodes isolated from banana in Nanjing County were confirmed as M. graminicola. To our knowledge, this is the first report of a natural infection of banana with M. graminicola. References: (1) A.M. Golden and W. Birchfield. Plant Dis. Rep. 52:423, 1968. (2) T. C. Vrain et al. Fund. Appl. Nematol. 15:565, 1992. (3) J. Xu et al. Eur. J. Plant Pathol. 110:309, 2004.

摘要

香蕉(芭蕉属)是世界上最受欢迎的水果之一,中国是世界第三大香蕉生产国。根结线虫是全球香蕉的常见害虫,但迄今为止,尚未有关于禾本科根结线虫对该作物造成危害的报道。在对中国福建省感染香蕉的根结线虫种类进行调查期间,2013年5月从南靖县的两个商业种植园采集了天宝蕉(AAA)的初级和次级根肿胀、形成虫瘿的样本。受影响的植株地上部分未表现出明显症状。严重感染的根畸形且开裂,组织变色并腐烂。对有症状的根进行检查发现,每个虫瘿内有一至数条禾本科根结线虫雌虫,卵块通常完全嵌入虫瘿内,不突出于根表面,且在虫瘿内孵化出第二代幼虫(J2)。该线虫的种群密度为每5克鲜根中有452至2056个卵和J2。很少观察到雄虫。对25条雌虫和20条J2的形态测量结果与禾本科根结线虫的原始描述相符(1)。雌虫的会阴花纹背腹呈卵形,背弓低至中等高度且圆润,无明显侧线;条纹平滑,部分在花纹背部被一些明显的不规则之字形条纹打断;尾感器靠得很近(13.1至19.7微米)。J2的尾部细长(63.4至75.5微米),有长而窄的透明区(13.1至19.9微米),末端明显呈棒状。从一条成熟雌虫中提取DNA。用V5367/26S(TTGATTACGTCCCTGCCCTTT/TTTCACTCGCCGTTACTAAGG)(2)扩增ITS1-5.8S-ITS2 rDNA区域,用C2F3/MRH106(GGTCAATG TTCAGAAATTTGTGG/AATTTCTAAAGACTTTTCTTAGT)(3)扩增COII和IRNA线粒体DNA基因,然后进行测序。将序列使用BLAST在数据库中搜索以验证其身份。ITS区域的序列为788 bp(GenBank登录号KM111531和KM236560),与Genbank中已知的禾本科根结线虫序列的同源性为96.8%至99.1%。线粒体DNA的序列为666 bp(KM111533和KM236559),与已知的禾本科根结线虫序列(KJ139963和HG529223)的同源性为99.1%至99.4%。在温室试验中,将约20厘米高的香蕉苗(天宝蕉)移栽到直径25厘米的花盆中,用每个采集的禾本科根结线虫种群的5000条J2进行接种,重复6次;设置未接种的对照。15周后,所有接种的植株生长受阻且黄化。根上的虫瘿症状与田间相似,对有虫瘿的根组织进行解剖发现存在线虫的不同生活阶段,种群密度为每5克鲜根中有1238至6562个卵和J2。未接种的对照植株生长良好,根上没有虫瘿症状。这些结果证实了该线虫对香蕉的致病性。基于这些结果,从南靖县香蕉中分离出的根结线虫被确认为禾本科根结线虫。据我们所知,这是香蕉被禾本科根结线虫自然感染的首次报道。参考文献:(1)A.M. Golden和W. Birchfield。植物病害报告52:423,1968。(2)T.C. Vrain等人。基础与应用线虫学15:565,1992。(3)J. Xu等人。欧洲植物病理学杂志110:309,2004。

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