截获泰国两种滑刃属线虫的记述
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作者单位:

1.昆明海关技术中心 昆明 650228;2.思茅海关 普洱 665099;3.昆明长水机场海关 昆明 650211

作者简介:

李旻,女,高级农艺师;研究方向:植物检疫;E-mail:beetfish@126.com。

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基金项目:

中国海关总署应急科研项目(No. 2018IK055),昆明海关科研项目(No. 2023KM005);


Morphological and Molecular Characteristics of Two Intercepted Aphelenchoides Species From Thailand
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Affiliation:

1.Technology Center of Kunming Customs, Kunming 650228; 2.Simao Customs, Puer 665099; 3.Kunming Changshui Airport Customs, Kunming 650211, China

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    摘要:

    本文描述了从泰国进境的束花凤梨(Tillandsia fasciculata)植株和万代兰(Vanda subconcolor)根系中检疫截获的两种滑刃属(Aphelenchoides)线虫。为明确其分类地位,本研究从形态学和分子生物学两个方面对其进行了系统鉴定。形态观察结果表明,泰国束花凤梨滑刃线虫种群的形态特征与原始描述的假古氏滑刃线虫(A. pseudogoodeyi)美国佛罗里达草莓(Fragaria × ananassa)种群高度相似,仅体长与虫体最前端至食道-肠交接处长度的比值(b值)存在差异,其余测计值均重叠。泰国万代兰滑刃线虫种群的形态特征与原始描述的苜蓿滑刃线虫(A. medicagus)美国苜蓿(Medicago sativa)种群高度相似,仅雌虫体长与尾长的比值(c值)存在差异,其余测计值均相互重叠。基于近全长的18S rDNA和28S rDNA D2-D3区序列构建了贝叶斯系统发育树。分析显示,在18S rDNA贝叶斯系统发育树中,泰国束花凤梨滑刃线虫种群与其他已报道的假古氏滑刃线虫几个种群聚在很高置信度的独立分支上(后验概率值为1.00),而在28S rDNA贝叶斯系统发育树中,所有假古氏滑刃线虫种群聚在较高置信度的独立分支上(后验概率值为0.98)。泰国万代兰滑刃线虫种群与苜蓿滑刃线虫种群在18S rDNA和28S rDNA D2-D3区贝叶斯系统发育树中,均聚为高置信度的独立分支(后验概率值均为1.00)。综合形态学和分子生物学证据,确定泰国束花凤梨滑刃线虫种群为假古氏滑刃线虫,而万代兰滑刃线虫种群为苜蓿滑刃线虫。这两种线虫均为泰国地理分布新记录种,同时束花凤梨和万代兰也分别是这两种线虫的新寄主记录种。本研究丰富了我们对滑刃属线虫生态、地理以及寄主范围等方面的认识,同时也为针对从泰国进境植物制定有效的植物保护和植物检疫措施提供了科学依据。

    Abstract:

    [Objectives] Two species of foliar nematodes were extracted from the leaves of Tillandsia fasciculata and the root rhizosphere of Vanda subconcolor, both of which were imported from Thailand, using a modified Baermann funnel technique. The taxonomic status of the two intercepted Aphelenchoides populations was determined through comprehensive morphological and molecular analyses. [Methods] For morphological analyses, nematode specimens were extracted, fixed, dehydrated and permanently mounted in anhydrous glycerin on glass slides. Specimens were photographed, measured and illustrated using a Leica DMi8 microscope. Morphometric measurements were carried out and analyzed in accordance with the de Man formula. For molecular analyses, genomic DNA was extracted from individual fresh nematodes. Two sets of universal primer pairs were employed for PCR amplification targeting both the 18S ribosomal DNA (rDNA) and the D2-D3 expansion segment of 28S rDNA. The resulting PCR products were subjected to separation, purification and sequencing. The newly obtained rDNA sequences were edited using DANMAN software package, compared against the NCBI database via BLAST, and subsequently deposited in the NCBI GenBank database. Phylogenetic analyses were conducted using a suite of bioinformatics tools:PhyloSuite v1.2.3 for extracting related species sequences and identifying the best-fit evolutionary model, MAFFT v7.313 for sequence alignment, MrBayes 3.2.7 for constructing Bayesian inference (BI) phylogenetic trees, and iTOL v6 for tree visualization. [Results] The population densities recovered from T. fasciculata and V. subconcolor were approximately 10 ind per 100 g leaves or roots, respectively. Both populations exhibited morphological similarities to A. besseyi, an economically significant plant-parasitic nematodecharacterized by a star-shaped tail tip and classified within the Aphelenchoides Group 3 as defined by Shahina (1996). Comparative morphological and morphometric analyses revealed that the nematode population from T. fasciculata closely resembled the Fragaria × ananassa-associated populationof A. pseudogoodeyi from Florida, USA, exhibiting overlapping ranges for all morphometric data except the b ratio. Similarly, the nematode population from V. subconcolor demonstrated morphological characteristics consistent with the Medicago sativa-associated population of A. medicagus from the United States, with overlapping measurements for all parameters except the c ratios in females. Phylogenetic analyses based on 18S rDNA and the 28S D2-D3 rDNA sequences further demonstrated the distinct evolutionary relationships among the two Aphelenchoides populations. In the 18S BI tree, the population from T. fasciculata formed a well-supported independent subclade (posterior probability, PP = 1.00) with A. pseudogoodeyi, while in the 28S BI tree, it formed a moderately supported independent subclade (PP = 0.98). Concurrently, the population associated with V. subconcolor clustered robustly with A. medicagusin in both the 18S and 28S BI trees, resulting in well supported independent subclades (PP = 1.00). [Conclusion] Based on an integrated analysis of morphological and molecular evidence, the nematode population derived from T. fasciculata was identified as A. pseudogoodeyi, whereas the population from V.subconcolor was identified as A. medicagus. Both of these nematode species represent new geographical records for Thailand, while T. fasciculata and V.subconcolor also serve as new host record species for the two nematodes, respectively. This study significantly enhances our understanding of the ecological characteristics, geographical distribution, and host range of Aphelenchoides nematodes. Furthermore, it offers a scientific foundation for formulating effective plant protection strategies and phytosanitary measures, aimed at mitigating risks associated with importation of plants from Thailand.

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引用本文

李旻,周绍芳,周剑,雷屈文,杨艳,张晔,杜宇.2025.截获泰国两种滑刃属线虫的记述.动物学杂志,60(3):399-413.

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  • 收稿日期:2024-07-08
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  • 在线发布日期: 2025-06-23
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