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1.
 水稻潜根线虫(Hirschmanniella oryzae)在水稻产区普遍存在,是水稻(Oryza spp.)的主要植物寄生线虫,在世界范围内造成产量损失。本研究通过比较GenBank数据库中潜根属线虫相关序列,以水稻潜根线虫ITS-rDNA序列为靶标,设计了水稻潜根线虫LAMP的一组特异性引物,包括一对外引物(F3:5′-ATCTTGTCCTTTGGCACG-3′,B3:5′-CGGTTGAACAAACAACGT-3′)和一对内引物 (FIP: 5′-CAGCATAGCAACAGAATGAATTCACGGTCGTAAACCTAATACGCG-3′,BIP:5′-TTGTACTACAATGGATTGTTTTCGCCTGATCCATCCACCCATG-3′);用琼脂凝胶电泳法和SYBR Green I染色法筛选合适的引物和扩增条件,建立了优化的水稻潜根线虫LAMP检测条件:扩增温度为57℃,反应时间为45 min,反应体系中dNTPs 和 MgSO4的浓度分别为1.4 mmol·L-1和7 mmol·L-1。本研究建立的检测方法可以检测鉴定水稻潜根线虫不同发育时期虫态(雌虫、雄虫、幼虫)个体,还可以从近缘种和其他植物寄生线虫混合的样品及水稻根组织样品中直接检测出水稻潜根线虫,且检测灵敏度达到1/1 000条成虫DNA。  相似文献   

2.
 为了提高链霉菌菌株JH108-2的发酵滤液对植物寄生线虫的毒力,采用正交试验设计,优化其发酵培养基。使用优化培养基[大豆粉20g、(NH4)2SO4 6g、蛋白胨3g、葡萄糖40g、酵母粉6g,NaCl 2g、K2HPO4 0.6g.CaCO3 2g、蒸馏水1000mL、pH 6.0]所得的发酵滤液,稀释10倍,处理南方根结线虫(Meloidogyne incognita)二龄幼虫,24h后校正死亡率达95.4%,比对照培养基的发酵滤液所产生的校正死亡率高6.9%,差异达到极显著水平(P=0.01)。  相似文献   

3.
香蕉穿孔线虫双重PCR快速检测技术研究   总被引:1,自引:0,他引:1  
 香蕉穿孔线虫(Radopholus similis)是国际上公认的植物检疫性有害生物之-, 是热带地区果树上最重要的线虫。通过设计的属水平上的特异性引物R1、R2(R1/2)和种水平上的特异性引物RS1、RS2(RS1/2)进行双重PCR扩增香蕉穿孔线虫, 可扩增出长度为362和291 bp的2个特异性片段谱带。同时分别对检测体系中反应参数进行优化, 最佳延伸温度为51.4℃, 引物R1/2与RS1/2最佳浓度配比为0.2μmol·L-1/1.2μmol·L-1, 并测试检测引物的灵敏度, 能进行有效检测的最低起始核酸量为100 pg;同时, 也能对单条线虫进行检测以及样品中线虫的检测。专化性测试证明, 2对引物能有效地进行香蕉穿孔线虫的快速分子诊断。  相似文献   

4.
南方、爪哇和花生根结线虫的快速灵敏的PCR鉴定方法   总被引:7,自引:0,他引:7  
 为了研制南方、爪哇和花生根结线虫快速灵敏的检测和鉴定方法,分别分离了4个南方根结线虫和3个爪哇根结线虫特异性的随机扩增多态性DNA (RAPD)片段。在这些RAPD标记DNA序列的基础上,设计了多对SCAR PCR引物,并用源于国内外的南方、爪哇、花生、北方和象耳豆根结线虫群体验证其扩增特异性和灵敏度。最终确定了3对高效扩增的SCAR引物,它们组合使用可以可靠灵敏地鉴定南方、爪哇和花生根结线虫。3对引物的扩增灵敏度达1/3条的二龄幼虫、雄虫或雌虫,这表明本研究研制的PCR鉴定法可用于生产实践中土样和根样中3种根结线虫快速灵敏的鉴定。  相似文献   

5.
 促分裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)在线虫的生长发育等方面具有重要作用,本研究探索利用RNAi技术沉默南方根结线虫mapk-1基因来抑制线虫的生长发育。根据mapk-1基因的cDNA序列设计引物,通过体外转录合成约550bp的dsRNA对南方根结线虫的卵进行RNAi以沉默mapk-1基因。试验结果表明,将南方根结线虫的卵块浸泡在含有2mg/mL dsRNA的M9缓冲溶液中,24h后卵块孵化出的2龄幼虫数量明显多于对照组(无dsRNA),但孵化出的幼虫死亡率高达90%,而对照组的死亡率低于5%,说明mapk-1基因的沉默抑制了卵块的孵化和线虫的生长,同时将孵化的幼虫接种番茄,14d后番茄根部无根结产生,35d后无卵块产生;而浸泡72h后卵块孵化出的2龄幼虫几乎全部死亡,并且孵化的线虫数量明显少于对照。提取导入dsRNA的卵块的RNA进行半定量PCR分析,结果表明mapk-1基因的mRNA被降解。  相似文献   

6.
进口画眉草籽中首次截获剪股颖粒线虫   总被引:1,自引:0,他引:1  
 弯叶画眉草[Eragrosti scurvula(Schrad.) Nees]原产于非洲,常栽培作牧草或布置庭园。关于画眉草上的线虫种类,仅津巴布韦有Pratylenchuszeae Graham (1951)及南非有Afrinawevelli vanden Berg (1985)的记载,而美国则没有报道。  相似文献   

7.
剪股颖粒线虫(Anguina agrostis)是牧草、草坪上的重要线虫,该线虫在进口草籽中常以幼虫形态出现,与其它粒线虫属幼虫形态相似,很难快速鉴定到种.本实验通过对单条幼虫DNA进行提取和PCR扩增,获得其rDNA的ITS区域,测序后与基因库中的剪股颖粒线虫的rDNA的ITS区域序列进行比较,其结果与基因库中编码为AF396343.1的核酸序列吻合,从而鉴定为剪股颖粒线虫.此外,还使用5种限制性内切酶对PCR产物进行了酶切,获得了剪股颖粒线虫的ITS-RFLP指纹图谱.  相似文献   

8.
为快速、准确、稳定的鉴定南方、花生和爪哇根结线虫,利用已报道的南方和爪哇根结线虫的两对特异性引物,结合本研究设计的花生根结线虫特异性引物,通过优化PCR反应体系,建立了3种根结线虫的PCR检测方法。结果表明,该方法能够特异性扩增以上3种根结线虫,特征片段长度分别为399、335和670 bp,灵敏度达到单条2龄幼虫的水平。研究结果将为以上3种根结线虫的快速鉴定提供技术支持。  相似文献   

9.
根结线虫种群的线粒体DNA分析   总被引:7,自引:0,他引:7  
 在同工酶和形态学鉴定的基础上,利用引物#C2F3和#1108对42个根结线虫种群线粒体DNA (mtDNA)中的COⅡLrRNA间区域进行特异性PCR扩增,35个种群的扩增产物约为1.7kb,其中29个是南方根结线虫,6个是爪哇根结线虫;3个花生根结线虫种群的扩增产物约为1.1kb;1个种群的扩增产物约为0.7kb,为根结线虫属在中国的新记录种;3个北方根结线虫种群的扩增产物约为0.5kb。用单条2龄幼虫提取物作模板得到的结果与大量提取DNA作模板的结果相同。为了区分产生相同大小片段的南方根结线虫和爪哇根结线虫,用限制性内切酶HinfⅠ对扩增产物进行酶切,结果表明:所有供试的南方根结线虫都可以被HinfⅠ酶切,且产生约1.3和0.4kb的2个限制性片段;但供试的爪哇根结线虫种群不能被酶切。由此表明,利用mtDNA PCR及酶切实验可以作为快速而准确地鉴定常见根结线虫的方法。  相似文献   

10.
淡紫拟青霉发酵滤液对大豆胞囊线虫趋化性的影响   总被引:5,自引:0,他引:5  
 实验室条件下,研究了大豆胞囊线虫幼虫对大豆根和根浸出液的趋化性以及淡紫拟青霉代谢产物对其趋化性的影响。结果表明,大豆胞囊线虫二龄幼虫(J2)对大豆根存在着一定的趋化性,而淡紫拟青霉发酵滤液对J2存在明显的驱避性。在根浸出液、真菌滤液及其混合液剂处理下,大豆胞囊线虫J2在WA平板上靠近处理液的0~1cm区间中分布率存在极显著差异(P <0.0 1),而加样时间(液剂与线虫同时放置或提前2 4h处理)对其分布率没有显著影响。大豆幼根蘸取滤液后对线虫J2在该区间的分布与不经处理的幼根差异显著(P <0.0 5),证明淡紫拟青霉发酵滤液可以明显地降低线虫与大豆根的亲和力,并强烈抑制线虫对大豆根的侵染。  相似文献   

11.
Two infestations of Anguina tritici in Western Australia were investigated for the presence of Clavibacter tritici. Five strains of C tritici were isolated from each site and compared with strains from Egypt, India, Iran and Iraq by allozyme electrophoresis. The Australian strains were genetically homogeneous but distinct from the non-Australian strains. A. tritici galls were found to be colonized by C. tritici , a feature of C. tritici infection not previously reported. C. tritici strains from Australia adhered to A, tritici but not to Anguina funesta. No corynetoxins were detected in C. tritici-infected galls. The potential of non-toxigenic Clavibacter species, such as C. tritici , for biocontrol of Clavibacter toxicus , the species responsible for poisoning livestock, is discussed.  相似文献   

12.
维氏粒线虫(Anguina wevelli)是危害牧草及草坪草的一类重要植物病原线虫。本文利用MAXENT生态位模型预测了维氏粒线虫在我国的适生区域,并结合该虫在我国的寄主及我国的地理信息,得出了该线虫在我国的潜在分布区域,结果显示,该线虫在我国云南保山南部及临沧东北部地区高度适生,云南除迪庆、怒江、丽江、昭通、西双版纳、文山、红河外的大部分中度适生。预测结果运用ROC曲线(受试者工作特征曲线)验证,测试数据的AUC(areas under curve,曲线下面积)值为0.971,达到了较好的精度。做好进境草籽的检验检疫对于保护上述地区的畜牧业发展有重要意义。  相似文献   

13.
Anguina wevelli is a pathogenic grass parasitic nematode, however it is difficult to identify based simply on morphology. This study developed a loop-mediated isothermal amplification (LAMP) assay to detect A. wevelli. The LAMP method developed could specifically detect A. wevelli in 45 min, and the detection limit was 1/80000 of the total DNA extracted from a single juvenile (J2), an equivalent of 2.5 pg of DNA. This is the first report of the detection of Anguina spp. by using a LAMP-based method.  相似文献   

14.
Nematicidal activity of the essential oils of different plant species belonging to the families Labiatae (Ocimum basilicum, Ocimum sanctum and Mentha piperata), Myrtaceae (Callistemon lanceolatus and Eugenia caryophyllata) and Gramineae (Cymbopogon caesius) and their major monoterpenoidal constituents, linalool, eugenol, menthol, cineole and geraniol was determined against second-stage juveniles of seed-gall nematode (Anguina tritici), citrus nematode (Tylenchulus semipenetrans), root-knot nematode (Meloidogyne javanica) and pigeon-pea cyst-nematode (Heterodera cajani). The essential oil of E. caryophyllata and its major constituent eugenol, as well as linalool and geraniol, exhibited non-specific activity against all the four nematodes tested.  相似文献   

15.
ABSTRACT Dead spot (Ophiosphaerella agrostis) is a damaging disease of young 相似文献   

16.
A fluorescent Pseudomonas sp. (strain LEC 1) isolated from soil suppressed Septoria tritici by 88% and Puccinia recondita by 98% when applied to wheat seedlings 3 h prior to inoculation with the pathogens. Fractionation and analysis of two inhibitory compounds from cultures of strain LEC 1 on thin-layer chromatography plates yielded the phenazine antibiotics 1-hydroxyphenazine (phOH) and chlorora-phin. 1-hydroxyphenazine reduced the growth of Pythium aphanidermatum, Fusarium oxysporum f.sp. vasinfectum, Sclerotinia sclerotiorum and S. rolfsii on a denned medium, with greatest activity against P. aphanidermatum and least against S. rolfsii. Application of 160 mg/l phOH to wheat seedlings reduced S. tritici by 61% and P. recondita by 75%. Lower concentrations of the antibiotic were less effective. Growth of 5. tritici in liquid medium was reduced by phOH at 5–20 mg/l and prevented at 40 mg/l. When phOH was added to the medium 48 h after inoculation (logarithmic growth phase), growth was interrupted but later recovered with phOH concentrations up to 40 mg/l; there was no recovery at 60 mg/l. Chlororaphin inhibited the growth of S. tritici on malt agar medium.  相似文献   

17.
ABSTRACT Two models for predicting Septoria tritici on winter wheat (cv. Riband) were developed using a program based on an iterative search of correlations between disease severity and weather. Data from four consecutive cropping seasons (1993/94 until 1996/97) at nine sites throughout England were used. A qualitative model predicted the presence or absence of Septoria tritici (at a 5% severity threshold within the top three leaf layers) using winter temperature (January/February) and wind speed to about the first node detectable growth stage. For sites above the disease threshold, a quantitative model predicted severity of Septoria tritici using rainfall during stem elongation. A test statistic was derived to test the validity of the iterative search used to obtain both models. This statistic was used in combination with bootstrap analyses in which the search program was rerun using weather data from previous years, therefore uncorrelated with the disease data, to investigate how likely correlations such as the ones found in our models would have been in the absence of genuine relationships.  相似文献   

18.
麦蒲螨[Pyemotes tritici(LaGreze-Fossat et Montagne)]是黑沙蒿主要钻蛀性害虫沙蒿大粒象(Adosomus sp.)和沙蒿尖翅吉丁(Sphenopterasp.)的重要寄生性天敌。为明确麦蒲螨的寄主选择行为,本文分别研究了麦蒲螨对黑沙蒿4种钻蛀性害虫、同一寄主不同发育阶段、受害黑沙蒿根部以及害虫虫粪的选择行为。结果表明:麦蒲螨对4种害虫以及同种害虫不同龄期幼虫有明显的选择现象,其中,对沙蒿尖翅吉丁的趋向性相对最高,沙蒿蛀茎蛾次之,沙蒿大粒象相对较小;相对于寄主的初孵幼虫,麦蒲螨对其老熟幼虫和蛹的趋向性更为强烈;不同种类害虫幼虫对麦蒲螨的寄主定位起到重要作用,而不同种害虫的虫粪对麦蒲螨的寄主选择行为也有一定的作用。相比受害黑沙蒿而言,健康黑沙蒿对麦蒲螨有明显的驱避现象。研究结果为揭示麦蒲螨对黑沙蒿不同钻蛀性害虫的寄主偏好性,利用麦蒲螨进行黑沙蒿主要害虫的生物防治提供了理论依据。  相似文献   

19.
ABSTRACT Most barley cultivars are resistant to stripe rust of wheat that is caused by Puccinia striiformis f. sp. tritici. The barley cv. Steptoe is susceptible to all identified races of P. striiformis f. sp. hordei (PSH), the barley stripe rust pathogen, but is resistant to most P. striiformis f. sp. tritici races. To determine inheritance of the Steptoe resistance to P. striiformis f. sp. tritici, a cross was made between Steptoe and Russell, a barley cultivar susceptible to some P. striiformis f. sp. tritici races and all tested P. striiformis f. sp. hordei races. Seedlings of parents and F(1), BC(1), F(2), and F(3) progeny from the barley cross were tested with P. striiformis f. sp. tritici races PST-41 and PST-45 under controlled greenhouse conditions. Genetic analyses of infection type data showed that Steptoe had one dominant gene and one recessive gene (provisionally designated as RpstS1 and rpstS2, respectively) for resistance to races PST-41 and PST-45. Genomic DNA was extracted from the parents and 150 F(2) plants that were tested for rust reaction and grown for seed of F(3) lines. The infection type data and polymorphic markers identified using the resistance gene analog polymorphism (RGAP) technique were analyzed with the Mapmaker computer program to map the resistance genes. The dominant resistance gene in Steptoe for resistance to P. striiformis f. sp. tritici races was mapped on barley chromosome 4H using a linked microsatellite marker, HVM68. A linkage group for the dominant gene was constructed with 12 RGAP markers and the microsatellite marker. The results show that resistance in barley to the wheat stripe rust pathogen is qualitatively inherited. These genes might provide useful resistance against wheat stripe rust when introgressed into wheat from barley.  相似文献   

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