Castillo P, Gutiérrez-Gutiérrez C, Palomares-Rius J E, Cantalapiedra Navarrete C, Landa B B
Institute of Sustainable Agriculture (IAS), CSIC, P.O. Box 4084, 14080 Córdoba, Spain.
Plant Dis. 2009 Dec;93(12):1353. doi: 10.1094/PDIS-93-12-1353B.
Some commercial vineyards producing the 'Condado de Huelva' wine denomination of origin in Almonte, Bonares, and Rociana (Huelva Province), southern Spain, showed general decline in sandy soils in 2009. Disease surveys revealed severe infections of grapevine rootstock Richter 110 feeder roots and heavy soil infestations by a root-knot nematode (Meloidogyne sp.). Infected plants showed a general decline as the only visible aboveground symptom, but when roots were inspected, moderate to small galls on secondary feeder roots were detected. The Meloidogyne sp. population was extracted and quantified from soil and root samples as previously described (1) and identified by the female perineal pattern, esterase (Est) and malate dehydrogenase (Mdh) phenotypes, and sequencing and maximum parsimony (MP) analysis of the ribosomal DNA region D2-D3 of 28S (2,4). Morphology of the perineal patterns and measurements of the second-stage juveniles (J2s) matched those of the original description of Meloidogyne hispanica (3). Enzyme analysis revealed two slow and a medium Est bands, a strong band, and two additional weaker bands coincident with the S2-M1 and N3 Mdh M. hispanica phenotypes (2,4). D2-D3 sequences of all three populations sampled were 100% homologous (GenBank Accession No. GQ375158). Phylogenetic analyses with MP of those sequences placed the Meloidogyne sp. in a clade (100% support) that included all M. hispanica sequences available from the GenBank database (4). M. hispanica was first found in Seville Province, southern Spain, infecting rootstocks of Prunus spp. Its distribution has been confirmed worldwide on different agricultural crops. Thus, M. hispanica has been reported to be infecting grapevines in South Africa and Australia (4); however, to our knowledge, this is the first report of M. hispanica infecting grapevines in Europe. Our data suggest that M. hispanica may pose a threat for vineyard production in southern Spain since M. hispanica was found in 52.63 and 47.36% of soil and root samples, respectively, from 19 fields in 'Condado de Huelva', with nematode population densities ranging from 2.4 to 129.6 eggs and J2s per 100 cm of soil and 1 to 1,797 eggs and J2s per gram of fresh roots. Furthermore, genes that confer resistance to other common root-knot nematodes reported on grapevine in Europe may not protect against M. hispanica. References: (1) K. R. Barker. Nematode extraction and bioassays. Page 19 in: An Advanced Treatise on Meloidogyne. Vol. II, Methodology. K. R. Barker et al., eds. North Carolina State University Graphics, Raleigh, 1985. (2) P. R. Esbenshade and A. C. Triantaphyllou. J. Nematol. 22:10, 1990. (3) H. Hirschmann. J. Nematol. 18:520, 1986. (4) B. B. Landa et al. Plant Dis. 92:1104, 2008.
西班牙南部韦尔瓦省的阿尔蒙特、博纳雷斯和罗西亚纳等地，一些生产“韦尔瓦伯爵区”法定产区葡萄酒的商业葡萄园，在2009年出现了沙质土壤普遍退化的情况。病害调查发现葡萄砧木里氏110的须根受到严重感染，且根结线虫（南方根结线虫属）对土壤造成严重侵害。受感染的植株仅表现出地上部分普遍衰退这一明显症状，但检查根系时，在二级须根上发现了中度至小型的虫瘿。按照之前描述的方法(1)，从土壤和根系样本中提取并定量分析南方根结线虫属种群，通过雌虫会阴花纹、酯酶（Est）和苹果酸脱氢酶（Mdh）表型，以及对28S核糖体DNA区域D2 - D3进行测序和最大简约法（MP）分析来鉴定(2,4)。会阴花纹的形态以及二龄幼虫（J2）的测量结果与西班牙根结线虫的原始描述相符(3)。酶分析显示有两条慢速和一条中速的Est带、一条强带，以及另外两条较弱的带，与西班牙根结线虫的S2 - M1和N3 Mdh表型一致(2,4)。所采集的三个种群的D2 - D3序列100%同源（GenBank登录号：GQ375158）。对这些序列进行MP系统发育分析，将南方根结线虫属置于一个分支中（支持率100%），该分支包含从GenBank数据库中获取的所有西班牙根结线虫序列(4)。西班牙根结线虫最初在西班牙南部的塞维利亚省被发现，感染李属植物的砧木。其分布已在全球范围内的不同农作物上得到证实。因此，据报道西班牙根结线虫在南非和澳大利亚感染葡萄(4)；然而，据我们所知，这是西班牙根结线虫在欧洲感染葡萄的首次报道。我们的数据表明，西班牙根结线虫可能对西班牙南部的葡萄园生产构成威胁，因为在“韦尔瓦伯爵区”19个田地的土壤和根系样本中，分别有52.63%和47.36%发现了西班牙根结线虫，线虫种群密度范围为每100立方厘米土壤中有2.4至129.6个卵和J2，每克鲜根中有1至1797个卵和J2。此外，欧洲报道的赋予葡萄对其他常见根结线虫抗性的基因可能无法抵御西班牙根结线虫。参考文献：(1) K. R. Barker。线虫提取与生物测定。载于：《根结线虫高级论述》第二卷，方法学。K. R. Barker等人编著。北卡罗来纳州立大学印刷厂，罗利，1985年。(2) P. R. Esbenshade和A. C. Triantaphyllou。《线虫学杂志》22:10，1990年。(3) H. Hirschmann。《线虫学杂志》18:520，1986年。(4) B. B. Landa等人。《植物病害》92:1104，2008年。
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