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1.
《农业科学学报》2017,(8)
Plant-parasitic nematodes are very common on cereal crops and cause economic losses via reduction in grain quality and quantity. During 2014, 83 soil samples were collected from wheat and barley fields in 21 districts of 13 provinces across five regions(Central Anatolia, Marmara, Aegean, Southeast Anatolia, and Black Sea Region) of Turkey. Cyst-forming nematodes were found in 66 samples(80%), and the internal transcribed spacer(ITS) sequencing and species-specific PCR identified the species in 64 samples as Heterodera filipjevi, Heterodera latipons, and Heterodera avenae. The predominant pathogenic cereal cyst nematode was H. filipjevi, which was found in all five regions surveyed. H. avenae was only detected in Southeast Anatolia whereas H. latipons was detected in Southeast Anatolia and Central Anatolia. ITS-rDNA phylogenetic analyses showed that H. avenae isolates from China clustered with H. australis, and Turkish isolates were closely related to European and USA isolates of this species. H. filipjevi from Turkey and China were clustered closely with those from the UK, Germany, Russia, and the USA. The density of many of these populations exceeded or approached the maximum threshold level for economic loss. To our knowledge, this is the first report of H. filipjevi in Diyarbakir, Edirne, and Kutahya provinces, and the first report of H. avenae in Diyarbakir Province. These results exhibit the most rigorous analysis to date on the occurrence and distribution of Heterodera spp. in Turkey's major wheat-producing areas, thus providing a basis for more specific resistance breeding, as well as other management practices. 相似文献
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【目的】明确小麦禾谷类作物孢囊线虫（cereal cyst nematodes,CCN）江苏群体的种类组成及群体间遗传变异情况,为抗病品种的选育、利用以及病害综合防治提供依据。【方法】对江苏省的13个代表性CCN群体进行孢囊、阴门锥和2龄幼虫的形态观察和形态测计并与相似种进行比较;PCR扩增上述群体的rDNA-ITS区并克隆测序,构建基于ITS序列的系统发育进化树。【结果】通过形态观察和形态测计值的比较,所测定的江苏群体均与已报道的禾谷孢囊线虫（Heterodera avenae）中国群体的测计值相接近;系统进化关系分析显示CCN江苏群体、国内其它地区以及国外禾谷孢囊线虫群体均处于同一大的进化分子簇,且国内和国外群体分别处于不同的进化分支。【结论】所测定的13个江苏省CCN代表性群体均为禾谷孢囊线虫,各群体间形态及ITS序列变异较小,江苏省目前尚未发现菲利普孢囊线虫（H. filipjevi）。 相似文献
3.
在传统真菌形态学分类研究的基础上,运用遗传学、分子生物学研究手段,结合生物信息学方法对具有诱导植物增产抗病的蛋白激发子产生菌链格孢(Alternaria spp.)JH505菌株进行了分子系统学研究.克隆了该菌株长度为1 267bp的rDNA序列.结果表明,rDNA包含部分18S rRNA、28S rRNA序列及完整的5.8S rRNA、转录外间隔区(ITS1、ITS2)rRNA序列,与A.tenuissima、A.cinerariae、A.dianthi、A.brassicae、A.zinniae相应的rDNA分别具有100%、98%、97%、97%和96%的相似性.结合菌落形态、孢子特征以及rDNA分析.鉴定该菌株为细极链格孢菌(A.tenuissima). 相似文献
4.
基于SCAR标记的小麦禾谷孢囊线虫快速分子检测技术 总被引：1，自引：2，他引：1
【目的】建立从禾谷孢囊线虫及其近缘种的混合群体中检测禾谷孢囊线虫的快速分子检测方法。【方法】使用随机扩增多态性DNA（RAPD）和特征序列扩增区域（SCAR）标记的方法,运用PCR技术进行特异性检测。【结果】用本研究建立的SCAR标记快速分子检测体系对9种32个线虫种群进行检测,能够直接从混合线虫样品中检测出禾谷孢囊线虫。该检测方法对禾谷孢囊线虫的孢囊、2龄幼虫具有扩增能力,最低检出阈值为1/2000个孢囊,1/80头2龄幼虫。【结论】本研究设计的SCAR标记快速分子检测体系能够从禾谷孢囊线虫及其近缘种的混合种群中快速检测出禾谷孢囊线虫,且检测准确、灵敏度高。 相似文献
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为了解小麦孢囊线虫(Cereal cyst nematode,CCN)在安徽省的分布和主要麦区CCN的种类,以及群体间关系,调查了淮北等麦区小麦根系白雌虫,用"Zig-Zag"法在怀远等地采集土样,用Fenwick&Oostenbrink法分离土样中的孢囊并孵化得2龄幼虫,利用孢囊阴门锥形态及de-man法鉴定CCN种类,扩增ITS特征片段,用MEGA5.05构建系统发育树。结果表明,安徽省52%采样点检出CCN,淮河以北采样点均检出CCN,江淮麦区7%的采样点检出CCN,长江以南麦区采样点未检出CCN。发生在怀远、凤台、灵璧、界首和颍上麦区小麦根际CCN鉴定为禾谷孢囊线虫(Heterodera avenae),淮河以北发生H.avenae麦区占淮北小麦种植面积的41.6%,另外,怀远H.avenae群体和其他群体亲缘关系较远,群体间形态有一定差异,但和亲缘关系差异无一致性。 相似文献
6.
《农业科学学报》2017,(7)
Heterodera avenae(cereal cyst nematode, CCN) infects many cereal crops and causes serious yield losses worldwide. Interaction studies investigating H. avenae and its hosts are still in their infancy. In this study, a barley model plant, the Hordeum vulgare cultivar Golden Promise, was investigated for its potential as a candidate model host to study its interaction with H. avenae. CCN-infective juveniles were attracted by the root tips and gathered around the root elongation zones of Golden Promise on 0.7% water agar plates. The juveniles invaded the roots and developed successfully until maturation at 40 days after inoculation in sterile sand soil. The cryotomy and syncytium measurements indicated that the syncytia enlarged gradually throughout the development of the nematodes and caused the corresponding root regions to swell obviously. Quantitative real-time PCR analysis showed that the down-regulation of defence-related barley genes and up-regulation of development-related barley genes contribute to the understanding of compatible interaction between H. avenae and Golden Promise. Barley stripe mosaic virus(BSMV) virus-induced gene silencing(VIGS) can be used in the roots of Golden Promise. In conclusion, the Hordeum vulgare cultivar Golden Promise is a suitable candidate model host for interaction studies with Heterodera avenae. The studies presented above document the first CCN host that not only has published genome context but also be compatible to BSMV VIGS. 相似文献
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根据NCBI基因库中根结线虫属rDNA序列,设计通用引物M18s/M28s,对采自云南文山、砚山、马关、蒙自等地的4个三七病原根结线虫种群(Meloidogyne hapla)rDNA区段进行聚合酶链式反应(PCR)扩增,并将扩增到的目的片段克隆到pGEM-T载体上。对重组克隆进行测序和序列分析,结果表明:4个种群rDNA同源性为100%,ITS区序列长度为479 bp,其中ITS1序列长度为213 bp,5.8S序列长度为159 bp,ITS2序列长度为107 bp。根据此序列设计-对特异性寡聚核苷酸引物Mhf/Mhr,应用PCR技术,以北方根结线虫(Meloidogynehapla)、南方根结线虫(Meloidogyne incognita)、花生根结线虫(Meloidogyne arenaria)和爪哇根结线虫(Meloido-gyne javanica)全基因组DNA为对照,对三七病原根结线虫全基因组DNA进行特异性扩增。结果表明,该对引物能从供试的北方根结线虫和三七病原根结线虫种群全基因组DNA中扩增到462 bp长度的分子片段,而南方根结线虫、花生根结线虫和爪哇根结线虫则无扩增产物。该引物可用于三七病原根结线虫的检测。 相似文献
8.
二硫氰基甲烷与灭菌唑和吡虫啉的联合使用对小麦孢囊线虫的防治 总被引：1，自引：0，他引：1
在田间自然情况下测试2种主要成分为二硫氰基甲烷的新型种衣剂及其与常规种衣剂灭菌唑和吡虫啉的联合使用对小麦孢囊线虫的防治效果。播种前用种衣剂对小麦种子分别进行包衣处理。分别调查播种前、抽穗期和收获后土壤中的孢囊数量及单孢囊卵量的变化,同时对小麦抽穗期的株高和收获时的千粒重等进行测定。结果表明:测试种衣剂对小麦进行包衣处理后均对该处理小区的孢囊及单孢囊卵量有抑制作用,其中甘农Ⅲ号种衣剂单独使用和甘农Ⅲ号种衣剂+灭菌唑(0.5∶0.5)联合使用处理小区的单孢囊卵量减退率最高,防效分别达到37.59%和32.86%,实际增产率分别达到16.09%和26.40%;甘农Ⅲ号种衣剂+灭菌唑(0.5∶0.5)联合使用和甘农Ⅲ号种衣剂单独使用2个处理,对抽穗期白雌虫的抑制率最高,分别为72.27%和70.29%,对孢囊的防效分别达到51.61%和50.74%;甘农Ⅲ号种衣剂+灭菌唑(0.5∶0.5)联合种衣剂、甘农Ⅲ号种衣剂+吡虫啉(0.5∶0.5)联合种衣剂2个处理的小区增产效果显著高于其他处理,理论增产率分别达到40.35%和44.39%。甘农Ⅲ号种衣剂+灭菌唑(0.5∶0.5)联合种衣剂可有效防控小麦孢囊线虫病害,不仅降低了农药的使用量,同时也显著提高了小麦产量,适于在生产上推广应用。 相似文献
9.
麦类作物对燕麦胞囊线虫抗病性遗传研究进展 总被引：3，自引：0，他引：3
郑经武 《沈阳农业大学学报》2001,32(3):224-227
概述了燕麦胞囊线虫致病型、麦类人地燕麦胞囊线虫抗病必一鉴定方法，介绍了迄今世界上在麦类作物对禾谷囊线虫抗病育种中已选育和生产上正推广的一些主要抗病基因（品种），阐述了中国燕麦胞囊线虫致病型（特别对rDNA中ITS区的PCR－RFLP分析，所获得的一个新的ITS类型－ITS C)以及麦类作物品种抗性的研究现状，这对中国开展针对燕麦胞囊线虫的抗病育种及相关的研究有重要的意义。 相似文献
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调查了湖北省禾谷孢囊线虫(Heterodera avenae)病的分布。在襄阳市重病田采样观察了禾谷孢囊线虫不同龄期的形态,研究了不同龄期线虫的发生动态。结果表明,禾谷孢囊线虫在襄阳、天门、潜江、钟祥、汉川都有发生,以襄阳最多,平均每100 mL土样含有效孢囊12.5个。在小麦播种后20 d就有2龄幼虫侵入根部,播种后143 d根内幼虫数量达到顶峰。入侵的幼虫在根内定居取食为害,在播种后158 d发育成白雌虫而露出根外,在播种后174 d白雌虫数量达最高峰。 相似文献
12.
银杏核糖体DNA内转录间隔序列初步分析 总被引：1，自引：0，他引：1
为研究银杏Ginkgo biloba种内核糖体DNA内转录间隔（ITS）区序列的多样性，以具有代表性的10个银杏嫩叶样本为材料，提取基因组DNA后用特异性引物进行PCR扩增，并直接测定其ITS区序列。结果表明，银杏ITS区（内含5．8S）总长度为1224～1226bp，其中ITS-1长度为821—823bp，5．8S长度为162bp，ITS-2长度为241bp。在全序列范围内，可变位点有28个，占总核苷酸的2．3％，但简约信息位点仅6个，变异量仅有0．5％。银杏的不同样本之间ITS区序列表现出高度一致。 相似文献
13.
Wheat yellow dwarf disease (BYD), caused by different species of barley/cereal yellow dwarf viruses (B/CYDVs), is one of the most serious cereal diseases in China and the Czech Republic. Because genetic diversity of the virus directly influences disease epidemiology, the molecular diversity and population structure of 24 Chinese isolates and 16 the Czech Republic isolates of BYDV-PAV from different regions in two countries were analyzed by sequencing their coat protein (CP) and readthrough protein (RTP) domain (RTD) genes and comparing the sequences with six CP and 16 RTP sequences of BYDV-PAV isolates from the NCBI database based on nucleotide identity position, phylogenetic analysis and nucleotide diversity. Nucleotide identities between the Chinese and the Czech Republic isolates for the CP were 76.6–99.4%, 73.9–89.1% for RTD (ORF5), respectively. The Chinese and the other country isolates showed 74.7–99.2% nucleotide identity for RTP (ORF3+ORF5). Phylogenetic analysis of CP sequences showed that 20 Chinese isolates clustered in the same clade, but the other four Chinese isolates clustered in another clade with the isolates from the Czech Republic and other counties. The population of BYDV-PAV in China had greater nucleotide variability and was more divergent than that in the Czech Republic. Geographical and ecological factors but not hosts might contribute to the population differences in the two countries. 相似文献
14.
患病北极红点鲑的病原分离与鉴定 总被引：1，自引：0，他引：1
对2004年5月四川省某养殖基地养殖的北极红点鲑患病鲑进行了病理学检查和病原分离,从病灶处分离到细菌和真菌。分离到的细菌经API生化鉴定为杀鲑气单胞菌杀鲑亚种(Aeromonas salmonicidasubsp.salmonicida)。用引物AP1和AP2对纯化后的细菌进行PCR扩增,结果扩增出长度为421 bp的DNA片段,对扩增片段进行克隆、测序,用NCBI-BLASTn在GenBank中搜寻相似序列,结果与杀鲑气单胞菌各株A层蛋白部分编码基因有99%以上的序列同源性。用引物EUS-F和EUS-R对分离到的真菌进行PCR扩增,结果扩增出长度为755 bp的DNA片段,对扩增片段进行克隆、测序,用NCBI-BLASTn在GenBank中搜寻相似序列,结果与水霉属各株的DNA片段(包括18S核糖体RNA基因部分序列、内转录间隔区1全序列、5.8S核糖体RNA基因全序列、内转录间隔区2全序列和28S核糖体RNA部分序列)有93%-100%的同源性;而与属于丝囊霉菌属的EUS各株相对应的DNA片段仅有60%的序列同源性。因此判定所分离的真菌为水霉。杀鲑气单胞菌是引起疖疮病和溃疡病等病的病原菌,而水霉广泛存在于水中,当鱼受伤后,水霉孢子容易从患病部位侵入鱼体,加重感染。因此判断该病主要是由杀鲑气单胞菌引起,而水霉则引起继发感染。 相似文献
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对大鲵(Andrias davidianus)2个驯化种群的核DNA进行PCR扩增,获得大鲵核DNA上编码核糖体5.8S rRNA和28S rRNA基因的部分序列和完整的ITS2序列(606 bp)。运用DNA分析软件对大鲵2个驯养种群(重庆水产研究所长寿湖珍稀鱼类繁育中心及广汉珍稀鱼类养殖公司)进行了遗传多样性分析。结果显示,该序列平均T、C、A、G含量分别为15.6%、36.5%、12.5%和35.4%,其中G、C的含量C+G(平均为71.9%)显著高于A、T含量A+T(平均为28.1%)。所测序列中有271个碱基发生颠换,112个碱基发生转换,转换与颠换比值(Si/Sv)为0.41,颠换大于转换。10个体均为单倍型,单倍型多样度(H)均为1.000 0±0.126 0,平均核苷酸差异系数(K)为5.932 1±0.861 4,核苷酸多样性(Pi)为0.631 9±0.013 9。种群遗传分化度(Fst)为0.030 5,中性检验及聚类分析表明2个群体没有分化成单一的群体,2个驯养种群遗传多样性好。 相似文献
17.
Molecular and morphological characterization of stunt nematodes of wheat,maize, and rice in the savannahs of northern Nigeria 
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下载免费PDF全文 Sulaiman ABDULSALAM PENG Huan LIU Shi-ming HUANG Wen-kun KONG Ling-an PENG De-liang 《农业科学学报》2022,21(2):586-595
Stunt nematodes (Tylenchorhynchus spp.) are obligate migratory root ecto-parasitic nematodes found in the fields of many cultivated crops. These nematodes, with phyto-sanitary potential, are frequently ignored or misdiagnosed as pests, and this may pose a threat to food security. The accuracy of its identification based on a morphological approach has been challenged recently, due to the overlapping of the morphological and morphometric characters of the species. Consequently, the objective of this study is to identify and characterize stunt nematodes present in 54 fields cultivated with cereal crops (wheat, maize and rice) in the savannahs of northern Nigeria, using integrative taxonomy and molecular approaches. The molecular and morphological studies identified and confirmed the presence of T. annulatus as the occurring specie in the savannahs of northern Nigeria. The phylogenetic analysis was carried out using the internal transcribed spacer (ITS) and 28S genes of ribosomal DNA further confirmed the presence of T. annulatus. The first molecular characterization and sequences of the ITS and 28S rDNA gene for T. annulatus from Nigeria were provided by this research. Also, according to our literature search, this is the first report on T. annulatus in wheat, maize and rice in the savannahs of northern Nigeria. Further study to test the pathogenicity of the parasitic nematode species found in this survey is recommended for the prioritization and development of efficient management strategies. 相似文献
18.
3个地理居群泥蚶核糖体DNA ITS区的RFLP分析(英文) 总被引：2，自引：0，他引：2
利用PCR-RFLP方法对3个地理居群泥蚶的核糖体DNAITS区进行分析,从分子水平探讨三者的遗传多样性及亲缘关系。结果表明:在选用的6种限制性内切酶中,BstHHI、HaeIII和Sse9I为多态性的限制性内切酶,MspI、RsaI和TaqI为单态性限制性内切酶。对酶切图谱进行数理统计后发现宁波海区泥蚶群体内核苷酸多样性指数π值为0.0017,温州海区泥蚶π值为0.0016,而莆田海区泥蚶π值为0.0011,说明宁波海区泥蚶群体内遗传多样性最大,温州海区群体内遗传多样性次之,莆田海区群体内遗传多样性最小。对所有酶切图谱的复合单倍型进行统计,共发现10种复合单倍型,其中宁波海区泥蚶群体内所含的复合单倍型类型最多为7种,其他2个海区均为6种,进一步证实宁波海区泥蚶群体内遗传多样性最大。另外,通过比较发现每个地理居群都有其特有的复合单倍型,如ABBABA型和BAABAA型复合单倍型是宁波海区泥蚶样本特有的,AAABBA型复合单倍型是莆田海区泥蚶样本特有的,温州海区泥蚶样本则具有AAACAA、BCAABA2种特有的复合单倍型。依据3个地理居群泥蚶间的净遗传距离Pnet值及聚类分析结果可知,莆田与温州海区泥蚶间亲缘关系最远(Pnet=0.0008),宁波与温州海区泥蚶间亲缘关系最近(Pnet=0.0003),宁波与莆田海区泥蚶间亲缘关系介于以上二者之间(Pnet=0.0004)。 相似文献
19.
用SDS-蛋白酶K裂解法(SDS-PK)、TE-蛋白酶K裂解法(TE-PK)、PCR buffer-蛋白酶K裂解法(PCRB-PK)和盐酸胍裂解法(Gu.HCl)提取禾谷孢囊线虫单个孢囊基因组DNA,以提取得到的基因组DNA为模板PCR扩增核糖体DNA(rDNA)片段,以扩增成功率和效率来反映基因组DNA提取的质量.结果表明:在所选的4种方法中,SDS-PK法扩增成功率最高,达85.71%,Gu.HCl法次之,TE-PK法最低.经微波裂解后,4种方法的DNA提取的成功率均有提高,其中微波裂解SDS-PK法为禾谷孢囊线虫单胞囊DNA提取的最优方法,PCR成功率可达96.97%. 相似文献
20.
松材线虫和拟松材线虫的PCR快速检测 总被引：8，自引：0，他引：8
利用PCR技术对松材线虫Bursaphelenchus xylophilus与拟松材线虫B．mucronatus的rDNA的ITS1区和5．8S区核甘酸序列扩增．根据松材线虫与拟松材线虫的ITS1序列区别,分别设计出松材线虫和拟松材线虫的特异性引物,实现了单条活的或FG(φ为4％的甲醛)固定的松材线虫和拟松材线虫的快速检测． 相似文献