Donald P A, Holguin C M, Agudelo P A
ARS USDA Crop Genetics Research Unit, Jackson, TN 38301.
School of Agricultural, Forest, and Environmental Sciences, Clemson, SC 29634.
Plant Dis. 2013 Oct;97(10):1389. doi: 10.1094/PDIS-09-12-0881-PDN.
The lance nematode Hoplolaimus magnistylus Robbins 1982 (3) was found for the first time in Tennessee in a cotton (Gossypium hirsutum L.) field (35°19.550' N, 89°24.535' W) in Fayette County in May 2011. In June of the same year, the species was also found in soil samples collected from a corn (Zea mays L.) field (36°15.736' N, 88°51.121' W) and a soybean (Glycine max L.) field (36°15.616' N, 88°51.118' W) in Weakley County, TN. Nematodes were extracted from the soil with a semi-automatic elutriator and further processed by sugar flotation-centrifugation. Population densities were between 30 and 50 individuals per 100 cm of soil in areas with noticeable stunting. Helicotylenchus sp. and Pratylenchus sp. were also present at less than 10 individuals per 100 cm of soil. Ten soybean seeds, cv. Hutcheson, were planted and inoculated with 50 H. magnistylus per 100 cm in steam-sterilized soil, and were maintained in a greenhouse. Forty-five days later, soybean plants exhibited at least one of the following symptoms: stunting and chlorosis, reduced root growth, and localized root lesions. Individual nematodes were handpicked and identified under a compound light microscope as H. magnistylus based on morphological and morphometric characteristics. The main diagnostic character for this species is the size of the stylet. In the populations collected, females had stylets ranging from 49 to 58 μm (mean 56 μm). Males and females were observed with head distinctly set off and massive cephalic framework, stylet long and robust and stylet knobs pointed anteriorly. The lateral field was areolated and had four incisures most of the body length, the excretory pore was prominent and located 190 μm (175 to 200 μm) from anterior end, hemizonid was large and located one or two annules posterior to the excretory pore, phasmids were large, conspicuous, and variable in position, and vulva was prominent and near midbody. This species is most similar to the more commonly reported H. galeatus, but differs from it in the longer stylet. Total DNA was extracted from single adults from each soil sample and the species-specific primers Hoc-1f (5'-AACCTGCTGCTGGATCATTA-3') and HM-3r (5'-AGACTGGACGGCCAAAGTT-3') designed by Bae et al. (1) were used to confirm the identification by amplification of a distinct 340-bp amplicon that differentiates this species from H. columbus, H. galeatus, H. concaudajuvencus, and H. stephanus. H. magnistylus was first described from soil samples taken from a corn and soybean field in Marianna, AR (3), and has been reported in association with soybean and corn in Louisiana and Mississippi (4). Robbins (4) reported that H. magnistylus was not a serious pest of irrigated cotton in Arkansas, but there are no other pathogenicity studies published for soybean, corn, or non-irrigated cotton. Other lance species, mainly H. galeatus and H. columbus, have been reported to cause serious damage to cotton in the Carolinas and Georgia (3). Previously, H. galeatus was reported in Tennessee by Bernard (2). Lance nematodes feed as migratory endo- and ectoparasites and injure the growing points of roots, causing stunting of plants. Because so little is known about the pathogenicity of this nematode, it becomes relevant to add our records of its known distribution in field crops in the United States. To our knowledge, this is the first report of H. magnistylus in Tennessee. References: (1) C. H. Bae et al. Nematology 11:471, 2009. (2) E. C. Bernard. University of Tennessee Bulletin 594, 1980. (3) R. T. Robbins. J. Nematol. 14:500, 1982. (4) R. T. Robbins. J. Nematol. 30(4S):590, 1998.
2011年5月,在田纳西州费耶特县的一块棉田(北纬35°19.550',西经89°24.535')中首次发现了长矛线虫Magnistylus magnistylus Robbins 1982(3)。同年6月,在田纳西州韦克利县一块玉米田(北纬36°15.736',西经88°51.121')和一块大豆田(北纬36°15.616',西经88°51.118')采集的土壤样本中也发现了该物种。使用半自动淘洗器从土壤中提取线虫,并通过糖浮选离心法进一步处理。在有明显发育不良的区域,每100厘米土壤中的线虫种群密度为30至50条。螺旋线虫属(Helicotylenchus sp.)和短体线虫属(Pratylenchus sp.)也有出现,每100厘米土壤中的数量少于10条。种植了10粒哈奇森品种的大豆种子,并在蒸汽灭菌的土壤中每100厘米接种50条Magnistylus magnistylus线虫,然后置于温室中。45天后,大豆植株出现了以下至少一种症状:发育不良和黄化、根系生长减少以及局部根系病变。通过在复式光学显微镜下根据形态和形态测量特征手工挑选并鉴定单个线虫为Magnistylus magnistylus。该物种的主要诊断特征是口针的大小。在采集的种群中,雌虫口针长度为49至58微米(平均56微米)。观察到雄虫和雌虫头部明显分开,头部框架厚实,口针长且粗壮,口针瘤向前突出。侧区具网纹,在身体大部分长度上有四条侧线,排泄孔明显,位于前端190微米(175至200微米)处,半环孔大,位于排泄孔后方一或两个环纹处,尾感器大,明显,位置可变,阴门突出且靠近虫体中部。该物种与更常见的加氏长矛线虫(H. galeatus)最为相似,但口针更长与之不同。从每个土壤样本中的单个成虫提取总DNA,并使用Bae等人(1)设计的物种特异性引物Hoc - 1f(5'-AACCTGCTGCTGGATCATTA-3')和HM - 3r(5'-AGACTGGACGGCCAAAGTT-3'),通过扩增一个340碱基对的独特扩增子来确认鉴定结果,该扩增子可将该物种与哥伦布长矛线虫(H. columbus)、加氏长矛线虫(H. galeatus)、尾短长矛线虫(H. concaudajuvencus)和斯蒂芬长矛线虫(H. stephanus)区分开来。Magnistylus magnistylus最初是从阿肯色州玛丽安娜的一块玉米田和大豆田采集的土壤样本中描述的(3),并且在路易斯安那州和密西西比州的大豆和玉米田中也有报道(4)。Robbins(4)报道Magnistylus magnistylus在阿肯色州不是灌溉棉花的严重害虫,但没有关于大豆、玉米或非灌溉棉花的其他致病性研究发表。其他长矛线虫物种,主要是加氏长矛线虫(H. galeatus)和哥伦布长矛线虫(H. columbus),已报道在卡罗来纳州和佐治亚州对棉花造成严重损害(3)。此前,Bernard(2)在田纳西州报道过加氏长矛线虫(H. galeatus)。长矛线虫以迁移性内寄生和外寄生方式取食,损伤根尖生长点,导致植物发育不良。由于对这种线虫的致病性了解甚少,增加我们在美国大田作物中其已知分布的记录就显得很有必要。据我们所知,这是Magnistylus magnistylus在田纳西州的首次报道。参考文献:(1)C. H. Bae等人,《线虫学》11:471, 2009。(2)E. C. Bernard,田纳西大学通报594, 1980。(3)R. T. Robbins,《线虫学杂志》14:500, 1982。(4)R. T. Robbins,《线虫学杂志》30(4S):590, 1998。