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百合脱毒组培技术研究 总被引：5，自引：0，他引：5
百合为百合科百合属(Lilium)多年生球根植物,是国际上主要鲜切花之一。百合在生产中一般采用无性繁殖,而百合病毒可通过种球传播。因此,病毒一旦感染百合,即在种球内不断积累,最终导致种质严重退化,给百合种植业造成巨大的经济损失。迄今已发现引起百合种球退化的病毒及其类似病原多达12种,其中危害最严重的有百合潜隐病毒(Lily symptomless virus,LSV),郁金香碎色病毒(Tulip breaking virus,TBV)和黄瓜花叶病毒(Cucumber mosaic virus,CMV)。随着组织培养技术的日臻成熟,通过组培脱毒为解决百合种球病毒性退化问题提供有效途径。国内外对百合组培脱毒进行的系列研究表明,获得脱毒苗的方 相似文献
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2004年4月27日，深圳检疫局盐田分局从荷兰进口的百合种球中抽取样品送动植中心进行病毒检测。经隔离种植后，发现有一株百合叶片上出现斑驳、条纹、扭曲等症状，疑是百合斑驳病毒侵染。随后，采用百合无症病毒RT-PCR检测方法，特异性扩增得到一条大约500bp左右的特异性条带，这与期望结果一致。为了保证检测结果准确无误，又将RT-PCR扩增产物进行测序，测序结果与GenBank上已登录的百合斑驳病毒多聚蛋白基因保守区序列一致，也与预先设计的基因片断完全一致。从而表明，从荷兰进口的百合种球中确实携带有百合斑驳病毒（Uly mottle virus）。 相似文献
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20 0 2年 7月 ,浙江某花卉公司从荷兰引进百合种球 1 3万个及介质土 ,所附的植物检疫审批单明确规定禁止携带草莓滑刃线虫Ａｐｈｅｌｅｎｃｈｏｉｄｅｓｆｒａｇａｒｉａｅ及穿刺根腐线虫Ｐｒａｔｙｌｅｎｃｈｕｓｐｅｎｅｔｒａｎｓ等危险性病虫。经检疫 ,发现百合种球及介质土携带大量植物寄生线虫 ,经进一步形态观察和测量 ,鉴定为我国植物检疫危险性线虫———草莓滑刃线虫Ａｐｈｅｌｅｎｃｈｏｉｄｅｓｆｒａｇａｒｉａｅ及穿刺根腐线虫Ｐｒａｔｙｌｅｎｃｈｕｓｐｅｎｅｔｒａｎｓ。这是宁波口岸首次截获上述两种线虫。现将草莓滑刃线… 相似文献
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进口百合种球中截获穿刺短体线虫 总被引：2，自引：1，他引：2
20 0 0年 7月至 9月 ,深圳某花卉公司经盐田口岸进口 8批产自荷兰的百合种球ＬｉｌｉｕｍＬ .,共计 1 70多万个种球 ,40多个品种。经检疫 ,发现其中有 6批 ,6个品种 ,8批次带有植物寄生线虫 ,后经进一步测量 ,鉴定为我国进境植物检疫三类危险性线虫———穿刺短体线虫Ｐｒａｔｙｌｅｎｃｈｕｓｐｅｎｅｔｒａｎｓ。这是深圳口岸首次从进口花卉中截获穿刺短体线虫。现将鉴定结果报道如下。1 材料与方法百合种球用改良贝尔曼漏斗法分离线虫 ,线虫经热的 4%甲醛杀死、固定 ,采用ＤｅＧｒｉｓｓｅ方法脱水 ,移到纯甘油中制成永久玻片 ,在Ｎｉｋ… 相似文献
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龙牙百合软腐病病菌——米根霉的分离鉴定及杀菌剂室内毒力测定初报 总被引：2，自引：0，他引：2
对低温储藏的部分腐烂的龙牙百合种球,进行了病原菌鉴定及致病性研究,并用菌丝生长速率法对10种杀菌剂进行了测定。经形态学和分子生物学鉴定,结果表明,导致龙牙百合种球腐烂的病原菌为米根霉(Rhizopus oryzae)。室内毒力结果显示,10种杀菌剂对米根霉菌丝的抑制效果有很大的差异。其中,1%申嗪霉素悬浮剂、30%己唑醇悬浮剂的抑菌效果较好,相应的EC_(50)分别为6.963、12.765 mg/L;抑菌效果最差是250 g/L嘧菌酯悬浮剂,EC_(50)为158 489.319 mg/L;30%己唑醇悬浮剂抑菌率最高,为100%。 相似文献
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以18S rRNA为内参照的多重RT-PCR检测3种百合病毒 总被引：5，自引：0，他引：5
本研究对多重PCR体系各成分和循环参数进行了摸索和优化,建立了以18SrRNA为内参照的同时检测3种百合病毒的多重RT-PCR体系,所检测的3种病毒是黄瓜花叶病毒(CMV)、百合斑驳病毒(LMoV)和百合无症病毒(LSV)。它们为侵染我国百合的主要病毒。在RT反应体系中加入3种病毒和18S rRNA的特异性反向引物的混合物,使反应体系中各反向引物终浓度均为0.5μmol/L,反转录酶(AMV)反转录合成各病毒和18SrRNA的互补第一链cDNAs。多重RT-PCR条件实验显示:将标准RT-PCR体系中的Taq HS DNA聚合酶量改为0.100U/μL,Mg2+浓度改为4.0mmol/L,各引物浓度选择0.2μmol/L,那么25个循环反应以上就能在一个反应管中以18SrRNA为内参照同时检测百合中的3种常见病毒。用该体系检测了10个百合样品,同时与32P同位素标记膜杂交检测作对照,结果显示,该检测体系检测灵敏度更高。 相似文献
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Wen-Chi Hu Chin-Hsing Huang Shu-Chuan Lee Chun-I Wu Ya-Chun Chang 《European journal of plant pathology / European Foundation for Plant Pathology》2010,126(1):43-52
Dasheen mosaic virus (DsMV), Turnip mosaic virus (TuMV), Konjac mosaic virus (KoMV) and Zantedeschia mild mosaic virus (ZaMMV) are important potyviruses previously identified in calla lily plants in Taiwan. In order to save time and cost of virus detection, a multiplex RT-PCR assay was developed for these calla potyviruses. Specific primers for each virus were designed based on the sequences of 3′ terminal region of respective viruses. To prevent false negative results, a primer pair specific to plant mitochondrial nad5 mRNA was used to produce a 185-bp fragment as an internal control of RT-PCR. The specificities of primers were confirmed by means of simplex and multiplex PCR assays. Optimal primer concentration ratio was identified by multiplex PCR assay. Total RNAs purified from virus-infected plants were used directly or mixed in different combinations, and then tested by multiplex RT-PCR. The result indicated that the expected RT-PCR products could be specifically amplified and identified on the basis of their molecular sizes. The detection sensitivity of multiplex RT-PCR was 25–625 times higher than that of indirect-ELISA (I-ELISA) depending on the virus. When applied to field surveys, multiplex RT-PCR could detect more single as well as mixed infection samples than I-ELISA. Accordingly, our multiplex RT-PCR assay provides a simple, rapid and reliable method for multiple potyvirus detection in calla lily. 相似文献
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百合黄瓜花叶病毒及其检测 总被引：3，自引：0，他引：3
应用酶联免疫吸附法(ELISA)检测了浙江丽水和杭州的东方百合、亚洲百合等栽培品种上黄瓜花叶病毒(CMV),64个田间样品中有26个带毒,检出率为40.6%;dsRNA检测结果与已知CMV-FQS株系的电泳条带相同,但百合组织中CMVdsRNA含量较低;电镜观察,受CMV侵染的样品中大多含有线状病毒,复合侵染率为35.9%;寄主反应测定显示从百合植株上获得的CMV株系均不能通过汁液摩擦接种侵染昆诺藜、苋色藜、普通烟、心叶烟等6科10种指示植物。 相似文献
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通过带病组织汁液和带病组织超薄切片负染电镜技术对侵染观赏百合的病毒粒子和内含体进行观察,证明在表现不同症状的5个品种中存在球状、短线状、长线状、短杆状4种病毒粒子和晶体状、风轮状、束状、卷筒状、正六边形5种内含体,通过ELISA检测及病毒粒子形态、大小及内含体形态分析,确定球状病毒粒子(直径29.5 nm)为黄瓜花叶病毒,短线状病毒粒子(593.29～639.97 nm×12.15～14.12 nm)为百合无症病毒,长线状病毒粒子(757.58～846.25 nm×12.12～12.75 nm)为马铃薯Y病毒,短杆状病毒粒子(500～807.69 nm×153.85～192.31 nm)为烟草脆裂病毒。 相似文献
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The denomination Tomato yellow leaf curl virus (TYLCV) comprises several viruses that cause severe damage to tomato crops in warm and temperate regions worldwide. TYLCV viruses are widespread in the Mediterranean Basin, in which two species have been reported: Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV, previously TYLCV-Is). The availability of methods convenient for the diagnosis of these viruses is essential. We have investigated several alternatives for reliable detection and differentiation of TYLCSV and TYLCV. Triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) proved to be very useful for large-scale diagnosis in field situations, but lacked discriminating capacity and sensitivity in the stages of infection in which low virus titre is present. The DNA-based methods are suited to laboratory operations and plant disease clinics, where accuracy of detection and discrimination of viruses is required. Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) was the most reliable method to discriminate between TYLCSV and TYLCV, but is not suited to high sample turnover. For large-scale testing, tissue print hybridization assay provides a reliable and sensitive alternative to PCR. 相似文献
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The multiplex RT-PCR approach was developed for simultaneous detections of Arabis mosaic virus(ArMV),Strawberry latent ringspot virus(SLRSV)and Lily symptomless virus(LSV)from the imported lily bulbs.The results indicated that good specificity and sensitivity for simultaneous detection were obtained.The ultimate of RNA detection with three viruses mixture was 305 pg.The ultimate of RNA detection with ArMV,SLRSV and LSV were 156.0,13.4 pg and 1.12 ng respectively.This approach has potential to be used in quarantine of imports and exports. 相似文献
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Nicola J. Spence Peter R. Mills Dez J. Barbara 《European journal of plant pathology / European Foundation for Plant Pathology》2000,106(9):843-847
Alstroemeria samples collected in the UK were tested for a range of viruses using ELISA. Alstroemeria mosaic virus (AlMV), alstroemeria carlavirus (AlCV), lily symptomless virus (LSV), cucumber mosaic virus (CMV) and tobacco rattle virus (TRV) were detected either singly or in combination in 67.5% of 203 samples. AlCV and LSV isolates from Alstroemeria and lily were studied and characterised serologically using existing antisera, and by PCR, using primers to an 11kDa open reading frame (ORF) unique to carlaviruses and to the coat protein gene of LSV. Sequences of isolates of AlCV and LSV from the coat protein gene were 94–99% similar and were 99% similar in the 11kDa ORF, supporting the view that these are strains of the same virus. 相似文献