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1.
粗茎秦艽Gentiana crassicaulis Duthie ex Burk.是一种重要的中药材, 近年来根结线虫病发生严重?为明确粗茎秦艽根结线虫病病原种类, 对其病原线虫进行种类鉴定, 以期为该病的防治提供理论依据, 采用雌成虫?2龄幼虫?会阴花纹形态特征及形态测量值进行形态学鉴定, 结合分子鉴定方法(比对rDNA-ITS和mtDNA-COⅠ序列?构建系统进化树, 序列特异性扩增区段比对)对病原线虫进行种类鉴定?结果表明, 该病原线虫雌成虫和2龄幼虫形态特征及测量值与北方根结线虫Meloidogyne hapla极为相似; 该病原线虫rDNA-ITS序列和mtDNA-COⅠ序列均与NCBI数据库中登录的北方根结线虫相似性达100%; 采用北方根结线虫特异性引物, 该病原线虫能扩增出1 500 bp的特异性条带, 与北方根结线虫特异性条带大小相同?上述结果表明, 云南省丽江市鲁甸乡粗茎秦艽根结线虫病的病原为北方根结线虫, 为首次发现北方根结线虫危害粗茎秦艽, 研究结果为进一步制定有效防治策略提供了理论依据?  相似文献   

2.
为确定四川省蓬溪县九叶青花椒种植区发病植株花椒根结线虫的种类, 进而为九叶青花椒根结线虫病防治提供依据, 本文根据根结线虫雌虫与2龄幼虫的形态特征及雌成虫的会阴花纹、雌虫酯酶同工酶图谱, 并结合特异性扩增及rDNA-ITS扩增, 根据ITS序列构建系统发育树, 对该地区九叶青花椒根结线虫病的病原进行了种类鉴定。结果表明该病原为南方根结线虫Meloidogyne incognita (Kofold & White) Chitwood。这是我国首次在九叶青花椒上发现南方根结线虫。  相似文献   

3.
四川省茂县花椒根结线虫病病原鉴定   总被引:4,自引:0,他引:4  
为明确四川省茂县花椒根结线虫的种类,通过根结线虫雌虫和2龄幼虫的形态特征及雌成虫的会阴花纹、雌虫酯酶同工酶及分子生物学检测(rDNA-ITS区和rRNA-IGS区),对花椒根结线虫进行了种类鉴定。三种方法的结果均表明在茂县地区花椒上发现的根结线虫为北方根结线虫(Meloidogynehapla),这是我国首次报道在花椒上发现北方根结线虫病,四川省是北方根结线虫新的分布区。  相似文献   

4.
为明确甘肃省党参根结线虫病的发生、分布及病原种类,在甘肃省党参主产区调查、统计田间根结线虫病发生情况,并通过形态学和分子生物学方法进行种类鉴定。结果表明,甘肃省6个党参主产区均有根结线虫病发生,病田率、病株率分别为40.7%、13.2%,病情指数为10.4。依据根结线虫各虫态的形态测量值和雌虫会阴花纹等特征,将危害党参的根结线虫初步鉴定为北方根结线虫Meloidogyne hapla Chitwood, 1949。rDNA-ITS区段和28S rDNA D2D3区段序列比对和系统发育树显示,6个根结线虫群体无核苷酸差异,与多个北方根结线虫聚为一支且有较高的置信度。利用特异性引物Mh-F/Mh-R扩增得到北方根结线虫的特异性片段,大小为462 bp。故综合形态学和分子生物学特征,将危害甘肃省党参的根结线虫鉴定为北方根结线虫。  相似文献   

5.
对河南省焦作市田间辖区种植的怀地黄根结线虫病发生情况进行了调查与研究,并采用作物根结线虫特异性(sequence characterized amplified region SCAR)标记引物对其病原根结线虫PCR扩增检测,结合雌虫会阴花纹的比较形态学和rDNA-ITS区序列测序分析的方法对怀地黄病原根结线虫进行种类鉴定。结果表明,我国焦作地区怀地黄的根结线虫为南方根结线虫[Meloidogyne incognita (KofoldWhite) Chitwood]。本研究首次明确了河南省焦作地区怀地黄病原根结线虫为南方根结线虫。  相似文献   

6.
通过形态学与分子生物学相结合的方法对引起甘肃省陇西县黄芩根结线虫病的病原种类进行了鉴定,并采用室内人工接种的方法测定其对黄芩的致病性。结果表明,从黄芩根系分离的根结线虫其形态特征及测量值与北方根结线虫Meloidogyne hapla基本一致。该线虫种群rDNA-ITS和28S rDNA D2/D3序列与NCBI数据库中的北方根结线虫序列相似性分别为99.60%和99.74% (rDNA-ITS: JX024147; 28S rDNA D2/D3: MW288147)。贝叶斯系统发育树显示,该线虫群体与其他北方根结线虫群体聚为一支,置信度在95%以上。利用北方根结线虫特异性引物Mh-F/Mh-R扩增并测序得到457 bp的特异性片段。因此,基于形态学结合rDNA-ITS、28S rDNA D2/D3序列和特异性片段长度,将黄芩根结线虫病病原鉴定为北方根结线虫M.hapla。接种该线虫后的黄芩生长缓慢,叶片黄化,40 d后须根上有明显的根结。经分离鉴定,致病群体为北方根结线虫。综上所述,甘肃省陇西县黄芩根结线虫病病原为北方根结线虫M.hapla,该线虫对黄芩具有较强的致病性,其繁殖系数为1.997。  相似文献   

7.
甘肃省保护地蔬菜根结线虫种类鉴定及其rDNA-ITS序列分析   总被引:2,自引:0,他引:2  
为明确甘肃省保护地蔬菜根结线虫的种类,采用rDNA-ITS-PCR方法结合其2龄幼虫、雌虫会阴花纹等形态学特征对采自甘肃省9个市的主要蔬菜生产基地保护地蔬菜根结线虫的11个种群样本进行了种类鉴定,并对其ITS区域进行序列测定及分析比对.结果表明:11个根结线虫种群的ITS区域序列与GenBank数据库已知南方根结线虫(Meloidogyne incognita)ITS区域序列的相似性达到99%,其ITS片段大小介于690~695 bp之间,表明甘肃省保护地中蔬菜根结线虫为南方根结线虫.  相似文献   

8.
宁夏葡萄苗圃根结线虫病发病症状、为害程度及种类   总被引:2,自引:0,他引:2  
为有效防治葡萄苗圃根结线虫病,采用拔根百分法和直接解剖法调查宁夏永宁县玉泉营镇葡萄苗圃葡萄苗木根结线虫病的发病症状、为害程度,并通过形态学和分子生物学对其种类进行鉴定。结果表明:15个葡萄品种的病害症状之间有一定差异。除抗砧3号外,其余14个葡萄品种均受根结线虫侵染,且不同葡萄品种受根结线虫为害的严重度差异明显,其中西拉、红芭拉蒂、爱神玫瑰、阳光玫瑰、马瑟兰、贝达和黑比诺上根结线虫病较严重,病情指数介于51.67~78.33之间,大青的病情指数最低,为21.67。不同葡萄品种的每根段雌成虫数量之间、每根段根结数量之间差异明显,其中葡萄品种贝达的每根段雌成虫数量和每根段根结数量最多,分别为12.47条和4.27个,葡萄品种黑比诺的每根段雌成虫数量和每根段根结数量较少,分别为3.60条和1.73个;每根段雌成虫数量随着每根段根结数量的增加而增加。根据根结线虫2龄幼虫、雌成虫和会阴花纹的形态学特征及分子生物学鉴定结果确定为害葡萄苗圃的根结线虫主要为南方根结线虫Meloidogyne incognita和花生根结线虫M. arenaria。除抗砧3号外,其它14个葡萄品种均不同程度的感染了南方根结线虫和花生根结线虫,且所有葡萄品种感染南方根结线虫的比例均较高。  相似文献   

9.
对湖南省蔬菜主产区的根结线虫病发生与危害程度进行了系统调查,结果显示蔬菜根结线虫病发生普遍,大棚蔬菜发病率高达80%,露地蔬菜发病率也在10%左右。对采自湖南省内10种蔬菜作物上的23个根结线虫种群进行了种类鉴定。采用2龄幼虫测量值、会阴花纹形态特征、PCR法及序列测定相结合的方法进行了鉴定,结果表明南方根结线虫是湖南省蔬菜根结线虫病的优势病原种群,占87%。  相似文献   

10.
对调入北京的草莓种苗根结线虫病的为害症状及病原进行了观察和鉴定,其症状为草莓种苗根部有明显而密集的根结,直径1~3 mm。通过根结长度百分数分级法明确根结指数为55.42;通过雌虫显微形态观察以及ITS序列测定,初步明确病原物为北方根结线虫(Meloidogyne hapla)。  相似文献   

11.
This study describes the development of species-specific pairs of PCR primers for the root-knot nematodes Meloidogyne chitwoodi, M. fallax and M. hapla that amplify species-specific RAPD fragments. After sequencing the fragments, longer primers were designed to complement the terminal sequences of the polymorphic DNA fragments. The resulting pairs of primers were used to generate the sequence-characterized amplified regions (SCARs). Using the developed pairs of SCAR primers, SCAR fragments of M. chitwoodi, M. fallax or M. hapla were easily amplified from DNA extracts from juveniles, egg masses, females of the particular nematode species investigated, either present alone, in a mixture with other nematode species or in infested plant material. A specially designed multiplex assay using three pairs of SCAR primers enabled the identification of multiple species in a mixture in a single PCR step. Single juveniles were easily identified by applying this multiplex assay followed by a subsequent multiplex PCR using three pairs of nested primers. The SCAR-PCR-based assays described have potential to be optimized for routine practical diagnostic tests. The usefulness of converting RAPD markers into SCAR markers is discussed.  相似文献   

12.
Organic amendments and green manure are potential alternatives to the harmful chemical control means currently used against plant-parasitic nematodes. In this work, Chrysanthemum coronarium was applied to the soil as a green manure to control the root-knot nematodes Meloidogyne incognita and M. javanica. Chrysanthemum coronarium significantly reduced nematode infection of tomato roots and improved plant-top fresh weight, both in the greenhouse and in microplots. Other green manures, derived from Anthemis pseudocotula, wild chickpea (Cicer pinnatifidum), Geranium spp. and wheat, were not as effective as C. coronarium. Chrysanthemum coronarium, retained its nematicidal activity even when applied as a dried material. Only mature C. coronarium plants, in their flowering stage, exhibited nematode control activity, but the green plant parts were more effective than the flowers. An aqueous extract of C. coronarium exhibited in vitro, nematostatic activity towards M. incognita and M. javanica second-stage juveniles and inhibited their hatching from eggs and egg-masses; its nematostatic activity was expressed also against other phytonematode species such as Heterodera avenae and Pratylenchus mediterraneus, but did not affect the beneficial entomopathogenic nematode Steinernema feltiae.  相似文献   

13.
南方根结线虫中国分离群体种内变异分析   总被引:1,自引:0,他引:1  
为调查我国不同地区和不同寄主上的根结线虫Meloidogyne spp.种类分布以及群体变异情况,基于酯酶和苹果酸脱氢酶同工酶图谱及SCAR分子标记技术对2017—2019年从6省19种植物根部组织分离到的40个根结线虫群体进行鉴定,针对南方根结线虫M.incognita群体分别通过寄主鉴别法进行生理小种鉴别,利用携带Mi抗性基因的番茄进行毒力测试,对2龄幼虫的口针长度和体长进行测量,并对核糖体ITS和线粒体Nad5基因序列进行比较分析。结果显示:根结线虫分离群体经鉴定包括38个南方根结线虫群体和2个象耳豆根结线虫M.erterolobii群体;38个南方根结线虫群体中有35个群体被鉴别为1号生理小种,其余3个群体被鉴别为2号生理小种;发现1个南方根结线虫群体CN19可在携带Mi抗性基因的番茄上侵染繁殖,为毒性群体,其余群体无法进行侵染和繁殖,为无毒群体。南方根结线虫群体2龄幼虫的口针长度和体长均差异较大,而不同寄主来源分离群体的ITS和Nad5基因序列也存在一定变异。基于ITS和Nad5基因序列构建的系统发育树将所有根结线虫群体归为南方根结线虫和象耳豆根结线虫组成的2个独立分支,但不能确定南方根结线虫不同群体的分子进化与其寄主来源和地理分布之间的相关性。  相似文献   

14.
Electron microscopic studies of the interaction of second-stage juveniles ofMeloidogyne hapla andArthrobotrys oligospora (CBS 289.82) strongly suggest that hyphae attachment to nematodes was mediated by a 0.1m thick layer matrix between the fungus and prey after contact with the nematode cuticle. The amorphous electron opaque matrix was irregularly distributed over the fungal surface, in some cases covering only the side attached to the nematode. Serial sections of adhesive hyphae showed that the extracellular matrix spread from the exact site of capture. A matrix with this thickness was never observed in the absence of nematodes.  相似文献   

15.
1,2-Dehydropyrrolizidine alkaloids (PAs), known to be nematotoxic in vitro, represent a class of secondary plant metabolites from hundreds of plant species worldwide. Pot experiments with the commercially available PA-containing plants Ageratum houstonianum, Borago officinalis, Senecio bicolor, and Symphytum officinalis demonstrate that Meloidogyne hapla is not per se repelled by these plants as all species were infested with nematodes. However, the development of M. hapla juveniles was completely suppressed on A. houstonianum and S. bicolor. Soil in which A. houstonianum and S. bicolor were cultivated and incorporated contained 200–400 times less nematodes than soil treated with Lycopersicon esculentum. Depending on their qualitative composition of PAs at least some of these plants thus appear to be valuable tools for integrated root-knot nematode management.  相似文献   

16.
Forty-six Meloidogyne populations from 14 provinces of China were characterised in terms of malate dehydrogenase, esterase phenotypes and HinfI restriction profiles of amplification products from the mitochondrial DNA (mtDNA) region between the COII and lrRNA genes. Isozyme phenotyping revealed that 29 of the populations were M. incognita, six were M. javanica, six were M. arenaria, three were M. hapla and two were M. enterolobii. HinfI restriction patterns of the COII-lrRNA region correlated with nematode isozyme phenotypes and enabled reliable differentiation and identification of the five root-knot nematodes occurring in China. The size and sequence of the mtDNA amplification product were determined for the first time for M. enteroloii, a potentially economically important crop pathogen. Sequence comparison showed that the sequence of the intergenic region between the COII and lrRNA genes for M. enterolobii was identical to that reported for M. mayaguensis. Together with published observations on morphology, host range and esterase phenotype of the two nominal species, the mtDNA sequence evidence suggests that M. mayaguensis could be conspecific with M. enterolobii.  相似文献   

17.
为探明长枝木霉Trichoderma longibrachiatum菌株TL16防治南方根结线虫Meloidogyne incognita的作用机理,采用原生质体转化法获得绿色荧光蛋白(green fluorescent protein,GFP)标记菌株GFP-TL16,通过测定菌株TL16和GFP-TL16对南方根结线虫卵和2龄幼虫(2nd-stage juvenile,J2)的寄生与致死作用,其发酵液对卵孵化的抑制作用和对J2的致死作用,以及菌株GFP-TL16在黄瓜根系的定殖情况和菌株TL16对番茄根结形成的抑制作用来综合分析其作用机理。结果显示:菌株TL16菌丝对南方根结线虫卵无寄生作用,处理19 d后卵降解率为26.33%,致死作用较低;菌株TL16分生孢子悬浮液处理南方根结线虫J2后72 h的致死率为1.65%,且无寄生作用。菌株TL16发酵液处理南方根结线虫J2后48 h的校正死亡率为10.71%,处理卵15 d后对卵孵化的相对抑制率为77.11%。菌株GFP-TL16可定殖于黄瓜根系中,经菌株TL16处理后接种南方根结线虫J2,番茄根结减退率为55.88%。表明长枝木霉菌株TL16可通过抑制根结线虫卵孵化和诱导番茄产生抗病性来防治根结线虫病。  相似文献   

18.
Agrobacterium tumefaciens (AT) is the causal agent of crown gall, a major problem in the family Rosaceae and particularly for Prunus spp. Crown gall symptoms result from the bacterial infection of the cells damaged mechanically at the collar or by root parasitic nematodes. Myrobalan plum (P. cerasifera) is susceptible to AT and is not a host for the root-knot nematode (RKN), M. hapla. Some clones of this plum carry single Ma resistance genes that control M. arenaria, M. incognita and M. javanica. The four above mentioned RKN and Myrobalan progenies segregating for Ma were used in experiments aimed at obtaining a better knowledge of the interaction between AT and RKN in relation to the RKN resistance genes. Prunus rooted cuttings, naturally infected with the bacterium were repotted, grown and inoculated individually with RKN. In a first experiment, Prunus plants were (i) either inoculated with 10,000 juveniles (J2s) of M. arenaria to provide a short inoculum pressure (SIP) or (ii) inoculated by association with one M. arenaria-galled tomato root system that produced a high and durable inoculum pressure of the same nematode species. Four months after RKN inoculation, plants were rated for nematode and bacterial root galling symptoms. RKN and AT galls were more numerous and more homogenous under DIP than under SIP. Nevertheless, for both inoculum regimes, AT galls were present in the RKN-susceptible clones (= carrying none of the Ma genes) and absent in the RKN-resistant clones. Subsequent experiments, conducted under DIP with M. arenaria, M. incognita, M. javanica and M. hapla, also showed, for the three first species, the presence of AT galls only in RKN-susceptible clones whereas Prunus plants inoculated with M. hapla and nematode-free controls were free of AT galls. Consequently RKN act as a wound agent in the AT infection process of Myrobalan plum only when the plant develops a compatible reaction (i.e. when it lacks the Ma resistance genes). Considering that J2s do penetrate the roots of resistant plants, the absence of crown gall symptoms on this material even under durable inoculum pressure strengthens the hypothesis that this nematode stage has a very weak effect on plant cells during the infection process. This is the first evidence of the protective effect of a RKN resistance gene against the expression of root crown gall consecutive to RKN infection. The protective effect of Ma and presumably of other RKN resistance genes against AT is a strong argument for their introgression into Prunus and other Rosaceae or bacterium-susceptible crops.  相似文献   

19.
ABSTRACT High infection rates of wild olive (Olea europaea sp. sylvestris) feeder roots and soil infestation by a new root-knot nematode were found in sandy soil at Vejer de la Frontera (Cádiz), southern Spain. Morphometric traits and analyses of the nematode esterase electrophoretic pattern as well as of the internal transcribed spacer 1 (ITS1)-5.8S gene and D2-D3 fragment of the 28S gene of rDNA showed that specimens differed clearly from known root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected wild olive plants and in olive planting stocks (cvs. Arbequina and Picual) artificially inoculated with the nematode. However, the nematode did not reproduce in artificially inoculated chickpea, pea, and tomato. Because of the ability of this new nematode to infect wild and cultivated olives only, we suggest the common name, "Mediterranean olive root-knot nematode." The species is herein described and illustrated, and named as Meloidogyne baetica n. sp. The new root-knot nematode can be distinguished from other Meloidogyne spp. by (i) the perineal pattern, which is almost similar to that of M. artiellia, characterized by distinct inner striae forming two distinct longitudinal bands, extending throughout the perineum to just below the vulva; (ii) female excretory pore anterior to the level of stylet knobs, excretory pore distance from anterior end/length of stylet ratio extremely small (0.5 to 0.8); and (iii) second-stage juveniles with elongate-conoid tail. Phylogenetic trees derived from maximum parsimony analyses showed that M. baetica is closely related to M. artiellia, the cereal and legume root-knot nematode.  相似文献   

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