花生茎线虫（Ditylenchus arachis n.sp.）新种鉴定、生物学及快速检测技术

发布时间：2018-03-07 15:55

  本文选题：花生茎线虫　切入点：形态特征　出处：《福建农林大学》2016年博士论文　论文类型：学位论文

【摘要】：花生茎线虫病发现于山东、河北多个花生产区,为我国首次报道的花生病害。本文对花生茎线虫病的病原线虫种类进行了系统鉴定,研究了病原线虫的相关生物学和快速检测技术,取得了如下主要研究结果：1.危害我国花生的茎线虫为一新种,命名为花生茎线虫(Ditylenchus arachis n. sp.)。新种的鉴别特性：侧唇片明显突出,口针纤细,长8.4-10μm,侧区有6条侧线,交合伞包至尾端的68%-86%,尾长圆锥形；基于rDNA-ITS、28SD2D3及18S区的系统发育分析证实Ditylenchus arachis n. sp的新种地位；致病性接种试验表明D. arachis n. sp可侵染花生(Arachis hypogaea)但不危害甘薯和马铃薯,与腐烂茎线虫(Ditylenchus destructor)不同。2.描述了花生茎线虫病的症状。通过田间症状观察和接种试验,发病田花生植株高矮参差不齐,缺株断垄,发病中心呈块状分布。受茎线虫侵染的花生植株呈现矮化、黄化,甚至整株枯死等症状；植株地下部表现出根系生长不良、结荚少、固氮菌减少、果荚表面出现黑褐色腐烂斑、种皮褐变皱缩等症状。从受害花生组织如果荚、种粒、胚栓、根茎及根际土壤可以分离出茎线虫虫体。3.创建了花生茎线虫的人工培养技术,用长柄链格孢(Alternaria longipes)和胡萝卜愈伤组织在25-30℃恒温黑暗培养30-60d后可获得较大的线虫量。花生茎线虫的不同地理种群无繁殖力差异。4.初步查明了花生茎线虫的越冬和传播途径。病原线虫可侵染鳢肠、凹头苋、反枝苋、地锦、牛筋草、马唐和狗尾草等花生地杂草；花生茎线虫侵染花生果荚和果粒,以幼虫为主要越冬虫态。花生茎线虫在田间杂草寄主及病花生种荚上越冬,成为病害的初侵染源,病花生种荚是病害远距离传播的主要途径。5.构建了花生茎线虫的快速检测技术体系。设计了一对花生茎线虫特异性引物DF1(正向5'-GGGAAGGCGAAGCTAAGCTA-3')和DR1(反向：5'-AACTTAGAGGCCAACGAAGCC-3'),在ITS1区扩增出171bp的特异性片段。该引物的可靠性高、特异性强,可实现花生茎线虫的准确、灵敏及快速的检测。
[Abstract]:Peanut stem nematode was found in several flower producing areas of Shandong and Hebei, which is the first reported peanut disease in China. The species of peanut stem nematode were systematically identified in this paper. The related biology and rapid detection techniques of pathogenic nematodes were studied. The main results were as follows: 1. Stem nematode was a new species, named Ditylenchus arachis n. sp. sp. sp. the new species was named Ditylenchus arachis n. sp. sp. the distinguishing characteristics of the new species were as follows: the lateral lip was prominent. The oral needle was slender, 8.4-10 渭 m in length, there were 6 lateral lines in the lateral region, 68 -86 from the umbrella to the end of the tail, and the tail was conical. Phylogenetic analysis based on the rDNA-ITSN 28SD2D3 and 18s regions confirmed the new status of Ditylenchus arachis nsp. The pathogenicity inoculation test showed that D. arachis n.sp could infect Arachis hypogaea but not sweet potato and sweet potato, which was different from Ditylenchus destructor. The symptom of peanut stem nematode disease was described. The peanut plants in the diseased field are varied in height and height, lack of plants and ridges, and the center of the disease is distributed in blocks. The peanut plants infected by stem nematodes show symptoms such as dwarfism, yellowing, and even the whole plant withered. The underground parts of the plants show poor root growth and few pods. Nitrogen fixing bacteria decrease, fruit pod surface appears dark brown rotting spot, seed coat browning to shrink and other symptoms. From the affected peanut tissue if pods, seeds, embryo suppository, The stem nematodes were isolated from rhizosphere and rhizosphere soil. The artificial culture technique of peanut stem nematodes was established. Alternaria longipesae) and carrot callus were cultured at 25 鈩，

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