共查询到17条相似文献，搜索用时 140 毫秒
1.
烟蚜传播黄瓜绿斑驳花叶病毒的可能性及方式 总被引：1，自引：0，他引：1
为确定烟蚜体内携带的是否为黄瓜绿斑驳花叶病毒（CGMMV）,采用RT-PCR和序列测定方法对其进行鉴定。本研究通过采集湖南宁乡、郴州、浏阳三个地区的田间烟蚜,提取RNA后反转录成cDNA,并以cDNA作为模板,通过设计引物进行PCR扩增,摇菌后测序。测序结果显示,在烟蚜体内扩增出与黄瓜绿斑驳花叶病毒相似度为99%的RNA病毒。对包括该分离物在内的22个基因核苷酸序列进行同源性比较和系统进化树构建,并结合地域来源进行分析,结果表明烟蚜体内的黄瓜绿斑驳花叶病毒(CGMMV)来源于其携带的CGMMV。通过分析推测,烟蚜可传播黄瓜绿斑驳花叶病毒,且传毒方式为持久性。 相似文献
2.
【研究目的】建立快速、准确、简便的检测大豆种子上菜豆荚斑驳病毒的免疫捕获一步RT-PCR方法（一步IC-RT-PCR）。【方法】采用抗原捕获和一步RT-PCR相结合的方法,并通过反应条件优化,确定一步IC-RT-PCR的反应程序,同时对该方法的特异性及实际检测效果进行验证。【结果】成功建立了一步IC-RT-PCR检测菜豆荚斑驳病毒方法,能够从感染菜豆荚斑驳病毒的大豆种子上扩增出预期大小的特异性片段,而对其他病毒及健康大豆无特异性反应;应用该方法对不同产地大豆种子样品进行检测,结果从来自美国的大豆样品中检出菜豆荚斑驳病毒,检出率为50%。【结论】一步IC-RT-PCR方法可以快速、准确地检测大豆种子上的菜豆荚斑驳病毒,适用于口岸检验检疫。 相似文献
3.
4.
明确黄瓜绿斑驳花叶病毒在寿光地区黄瓜上的分布及危害，为寿光高品质黄瓜的生产提供参考依据。2017—2018年对寿光不同区域、品种、时间和生育期的黄瓜进行黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)系统跟踪调查，分析山东寿光黄瓜上CGMMV的分布及危害。所调查的10个乡镇均不同程度受CGMMV影响，其中稻田镇发病最为严重，发病率达49.64%；光皮黄瓜受害较带刺黄瓜严重；全年病毒高发期集中在5—8月，6月发病最为严重，发病率高达63.33%；黄瓜上发病最为严重的生育期是成熟期。分析原因除品种抗性差异外，温、湿度及个人管理模式等因素有利于CGMMV的发生和流行，因此，合理降温和加强管理可减轻CGMMV的危害。 相似文献
5.
本研究以黄瓜绿斑驳花叶病毒侵染的黄瓜叶片为材料,应用双抗体夹心酶联免疫吸附测定法（DAS-ELISA）、反转录-聚合酶链反应（RT-PCR）、核酸斑点杂交（NASH）技术,进行了检测灵敏度比较研究。三种检测方法灵敏度实验结果表明：RT-PCR检测灵敏度高于地高辛标记的核酸斑点杂交技术,以血清学为基础的DAS-ELISA检测灵敏度最低。通过灵敏度对比分析得出,核酸斑点杂交技术可以替代RT-PCR应用于基层的检疫检测。 相似文献
6.
7.
8.
为揭示黄瓜绿斑驳花叶病毒的分子流行学特征,通过RT-PCR和cDNA克隆技术获得并分析了从北京甜瓜和山东西瓜上分离的2个黄瓜绿斑驳花叶病毒(CGMMV)分离物CGMMV-Beijing和CGMMV-Shandong的全序列.结果表明CGMMV-Beijing和CGMMV-Shandong的基因组分别由6423和6427个核苷酸组成,包括5'及3'端非编码区和4个开放性的阅读框,分别编码129kDa和186kDa复制相关蛋白、29kDa的移动蛋白及17.4kDa的外壳蛋白.寄主范围测定显示在供试的6科11种植物中CGMMV-Beijing和CGMMV-Shandong均可侵染葫芦科的葫芦、南瓜和西瓜以及茄科的本氏烟和藜科的苋色藜,不侵染供试的其它植物.CGMMV-Beijing和CGMMV-Shandong基因组的核苷酸同源性为99.5%,186kDa蛋白的核酸和氨基酸同源性分别为99.7%和99.8%,移动蛋白的核酸和氨基酸同源性分别为99.3%和98.1%,外壳蛋白的核苷酸同源性为100%.多序列比对分析结果显示,CGMMV的各分离物之间的分子差异并不明显,核苷酸同源性均在98%以上.将CGMMV-Beijing和CGMMV-Shandong与侵染葫芦科植物的烟草花叶病毒属其它成员比较,结果显示其亲缘关系较远,基因组的核苷酸同源性介于56.7%～59.6%,外壳蛋白的氨基酸同源性在44.7%～54.0%之间.综合分析显示,CGMMV分离物之间的差异并不明显,可能具有共同的侵染来源. 相似文献
9.
利用RT-PCR对黄瓜病毒病毒原种类进行检测 总被引：1，自引：1，他引：0
根据黄瓜花叶病毒(CMV)的复制酶序列和西瓜花叶病毒2号(WMV-2)、烟草花叶病毒(TMV)、黄瓜绿斑花叶病毒(CGMMV)的外壳蛋白基因序列以及南瓜花叶病毒(SQMV)的运动蛋白序列设计、合成引物,以感病组织和健康组织总RNA为模板,进行cDNA合成和PCR扩增。对2002～2003年采集的98份黄瓜病毒病样本进行了检测,结果表明,天津地区黄瓜主要毒原种类有CMV,WMV-2和TMV,感染率分别为样本数的96．94％,77．55％和38．67％,CMV和WMV-2 2种病毒的复合感染率为76．53％,3种病毒的复合感染率达到14．67％。 相似文献
10.
山东葫芦科蔬菜上病毒病种类检测及黄瓜花叶病毒分离物的亚组鉴定 总被引：1，自引：0，他引：1
为了检测和鉴定山东地区葫芦科蔬菜上病毒病种类,从山东诸地11个蔬菜种植区采集了867个葫芦科蔬菜疑似感病样品。利用黄瓜花叶病毒、黄瓜绿斑驳花叶病毒、西瓜花叶病毒、烟草花叶病毒、小西葫芦黄花叶病毒、南瓜花叶病毒、甜瓜黄斑病毒、瓜类褪绿黄化病毒、李属坏死环斑病毒及甜瓜坏死斑点病毒等特异性引物对疑似感病样品分别进行RT-PCR检测,各病毒检出率分别为34.8%,10.4%,20.0%,41.7%,27.0%,7.8%,2.6%,1.7%,0,0.9%。表明除PNRSV外,其他9种病毒在山东各地均有发生,且CMV和TMV发病率较高,2种或2种以上病毒复合侵染也很普遍。同时为明确山东地区CMV分离物的株系分化状况,选取11个地区有代表性的CMV阳性样品,测定其外壳蛋白(Coat protein,CP)和RNA3核苷酸序列并进行序列比对和进化分析,结果显示侵染山东地区葫芦科蔬菜的CMV分离物均属于IB亚组,与韩国分离物As(AF013291)相似性最高,未发现其他株系。 相似文献
11.
Detection of SO_2 Residual in FLOS CHRYSANTHEMI by Accelerated Solvent Extraction/Ion Chromatography
《Medicinal Plant》2015,(Z2)
[Objective] To discuss the detection method for SO2 residual in FLOS CHRYSANTHEMI. [Methods] Accelerated Solvent Extraction / Ion Chromatography was used to detect the SO2 residual in FLOS CHRYSANTHEMI. Alkaline solution was used as the extracting solution of grinded sample. After extraction by accelerated solvent extraction apparatus and purification by C18 solid phase extraction,the sample was isolated by IonP ac AS9- HC ion chromatographic column with carbonate as the leacheate. Detection was carried out by electrical conductivity detector. [Results] SO2 had good linear relationship within the range of 0. 5- 50 mg / L( r2= 0. 999 7). The quantitative lower limit( LOQ) and and detection limit( LOD) were 0. 17 and 0. 05 mg/L,respectively. With blank sample as the matrix,the average recovery rates at high,middle and low dosages were 82. 9%- 92. 6% with RSD( n = 6) less than 5. 0%. [Conclusions]Compared with the traditional method,this method had the advantages of high extraction rate and automation degree,and was suitable for the large-scale detection of SO2 residual in FLOS CHRYSANTHEMI. 相似文献
12.
[Objective] The aim of this study is to establish an method for cotton metabolites identification by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) analysis, and to investigate the adduct types, dominant adducts and appropriate ESI ion modes of cotton secondary metabolites under the determined UPLC and ESI conditions. [Method] UPLC-ESI-MS was employed to analyze 18 cotton metabolite standards, Online XCMS software was used to extract the nontargeted mass spectrum data, and MATLAB software was used to prepare calculation programs for the identification method of cotton metabolite standards. [Result] A high-throughput identification method of cotton secondary metabolite, named POSid and NEGid separately for the positive and negative ESI modes, was established based on the calculated accurate molecular weight. In the determined UPLC condition and positive and negative ESI mode, 14 cotton metabolite standards were correctly identified. There found 6 adducts including [M+H]+, [M+Na]+, [M+NH4]+, [2M+NH4]+, [2M+Na]+ and [2M+H]+ in positive ion mode, and 8 adducts including [M-H]-, [2M-H]-, [M+Cl]-, [M+FA-H]-, [3M-H]-, [M+Na-2H]-, [M-H2O-H]- and [M+TFA-H]- in negative ion mode, while 1 to 6 adducts were observed in the mass spectrum of a single standard, and each metabolite standard had a dominant adduct of a preference for ESI mode. Melibose was suitable for ESI positive ion mode detection, gossypol was suitable for both ion modes detection; and 12 compounds were suitable for negative ion mode detection due to mass spectrum signals of their dominant adduct stronger in negative ion mode than positive ion mode. [Conclusion] Based on the accurate molecular weight, the established identifying method can identify the 18 cotton metabolite standards with their nontarget mass spectrometry data. Under the determined UPLC and ESI condition, the dominant adducts of cotton secondary metabolites have the preference of ESI mode. These results provide technical and theoretical data support for further study of cotton metabolomics. 相似文献
13.
旨在建立一种基于X射线荧光光谱法快速检测原料公干鱼(原料鱼)中镉的方法。应用X射线荧光光谱法快速检测原料鱼初始样及干燥样中的镉含量,对快速检测与标准检测结果的相关性进行了分析;考察了对原料鱼进行快速干燥处理的可行性;采用原料鱼的标准测定值对X射线荧光光谱仪进行校正后,考察了快速检测方法的精密度、检测限、准确度等性能参数。结果表明:X射线荧光光谱法能准确测定原料鱼干燥样中的镉,检测限达0.088 mg/kg,线性范围为0.088~1.286 mg/kg,相对标准偏差低于10%。该快速检测方法测定单个样品只需40 min左右,可满足现场对原料鱼中镉的快速测定要求,为防控原料鱼食品中镉超标风险提供了一种高效的筛查手段。 相似文献
14.
Cucumber green mottle mosaic virus (CGMMV) is a severe threat for cucumber production worldwide. At present, there are no cultivars available in the market which show an effective resistance or tolerance to CGMMV infection, only wild Cucumis species were reported as resistant. Germplasm accessions of Cucumis sativus, as well as C. anguria and C. metuliferus, were mechanically infected with the European and Asian strains of CGMMV and screened for resistance, by scoring symptom severity, and conventional RT-PCR. The viral loads of both CGMMV strains were determined in a selected number of genotypes using quantitative RT-PCR. Severe symptoms were found following inoculation in C. metuliferus and in 44 C. sativus accessions, including C. sativus var. hardwickii. Ten C. sativus accessions, including C. sativus var. sikkimensis, showed intermediate symptoms and only 2 C. sativus accessions showed mild symptoms. C. anguria was resistant to both strains of CGMMV because no symptoms were expressed and the virus was not detected in systemic leaves. High amounts of virus were found in plants showing severe symptoms, whereas low viral amounts found in those with mild symptoms. In addition, the viral amounts detected in plants which showed intermediate symptoms at 23 and 33 dpi, were significantly higher in plants inoculated with the Asian CGMMV strain than those with the European strain. This difference was statistically significant. Also, the amounts of virus detected over time in plants did not change significantly. Finally, the two newly identified partially resistant C. sativus accessions may well be candidates for breeding programs and reduce the losses produced by CGMMV with resistant commercial cultivars. 相似文献
15.
利用多重PCR技术同时检测6种棉花转化体的方法研究 总被引：1，自引：1，他引：0
【目的】建立1种转基因棉花多重聚合酶链式反应(Polymerase chain reaction,PCR)检测方法。【方法】试验选择6种转基因棉花作为多重PCR检测对象,通过引物终浓度配比和引物退火温度的两要素全组合优化多重PCR反应体系,并分析方法灵敏度。【结果】多重PCR较优的MON1445、GHB614、MON15985、MON88913、LLCOTTON25、MON531引物终浓度为0.25、0.30、0.25、0.16、0.30、0.20μmol·L-1,较优引物退火温度为56℃,方法灵敏度为66个拷贝棉花基因组。利用盲样对上述体系的验证结果显示,所有盲样扩增条带与其含有的转基因转化体完全一致。【结论】本研究建立的体系可用于6种棉花转化体的快速检测。 相似文献
16.
【目的】荧光原位杂交技术可以实现DNA序列直观准确的染色体定位,是基因组深入研究的重要技术之一。染色体特异探针的获得是该技术应用的关键。本研究旨在建立棉花寡核苷酸荧光原位杂交技术。【方法】利用已经公布棉花基因组序列数据,采用生物信息学方法获得染色体特异的寡核苷酸库,随后用乳化聚合酶链式反应方法标记成荧光探针,在棉花有丝分裂中期染色体上进行原位杂交。【结果】建立了一套棉花寡核苷酸荧光原位杂交技术体系。【结论】该体系可用于棉花单染色体识别鉴定。 相似文献
17.