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《中国植保导刊》2021,(7)
为建立苹果茎沟病毒(ASGV,Apple stem grooving virus)的有效脱除方法,以3个梨品种的离体植株为材料,比较了4种脱毒方法对ASGV的效果。结果显示,3个梨品种的离体植株在茎尖培养、茎尖病毒醚处理、茎尖变温热处理和茎尖病毒醚加变温热处理后,其平均脱毒率分别为50.0%、70.8%、69.6%和98.2%。茎尖病毒醚处理对植株生长的影响与单独茎尖培养基本一致,茎尖变温热处理和茎尖病毒醚加变温热处理对植株长势和增殖的影响与单独茎尖培养存在差异,但对植株总体生长状况和存活率的影响不大。结果表明,采用茎尖病毒醚加变温热处理脱除ASGV的效果最佳,可用于梨无病毒苗木的生产。 相似文献
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《中国植保导刊》2019,(9)
为探索建立一种简便高效的苹果砧木脱毒方法,提高脱毒苗成活率和脱毒率,本研究以常用苹果砧木系NY2和QD-V-2组培苗为材料,利用变温热处理结合茎尖培养的方法对苹果砧木所携带的ASGV、ACLSV、ASPV病毒进行脱除。结果表明,苹果砧木系NY2组培苗经变温热处理平均存活率为55.17%,经恢复培养的茎尖平均存活率为11.07%;QD-V-2组培苗经变温热处理平均存活率为57.96%,经恢复培养的茎尖平均存活率为15.98%。最终获得脱除ACLSV和ASGV的NY2脱毒苗4株,获得脱除ASPV和ASGV的QD-V-2脱毒苗4株。该方法操作简便,可应用于苹果无毒苗木的培育工作中。 相似文献
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Calavan等(1972)报道采用40/30℃ 44/30℃6 2星期,脱除北京柠檬碎叶病毒。Roistacher等(1972)采用50℃湿热处理北京柠檬3～22h,得到无碎叶病毒,并于1976、1977年报道,茎尖嫁接不能脱除柑桔碎叶病毒(CTLV)。宫川(1980)提出40/30℃120日脱除椪柑、蕉柑等碎叶病毒。Koizumi(1984)采用40/30℃9日 35/30℃13～20日 茎尖 相似文献
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类病毒是已知最小的植物病原物,可侵染蔬菜、果树以及花卉等多种农作物,并对农业生产造成严重影响。目前,对感染类病毒的植株进行脱毒处理是一种有效的防控措施,本文针对果树、蔬菜和花卉上几种常见的类病毒,综述了茎尖培养、热处理结合茎尖培养、低温处理、超低温处理、化学处理及不含叶原基的顶端分生组织再生等植物类病毒的脱除技术,分析了不同方法的应用效果及所适合脱除的类病毒种类,同时展望了类病毒病害在未来植物病害防控中的重要地位,并提出综合考虑各种因素、制定合理方案、选用适宜方法的类病毒脱除建议。 相似文献
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单独采用茎尖嫁接法和单独采用热处理法均难脱除温州蜜柑萎缩病毒(SDV),但采用白天40℃光照,夜间30℃黑暗(各12小时)热处量7~43天,结合茎尖嫁接法获得的72株茎尖苗全部脱除SDV。所以采用上述方法热处理7天,结合茎尖嫁接是脱除SDV的简便可靠的方法。 相似文献
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研究了植物诱抗剂3-丙酮基-3-羟基羟吲哚(3-acetonyl-3-hydroxyoxindole, AHO)联合茎尖培养从试管苗中脱除马铃薯S病毒(PVS)?马铃薯Y病毒(PVY)的方法和效率?取PVS侵染的‘定薯3号’和‘定薯4号’以及PVY侵染的‘靖薯3号’和‘靖薯4号’的壮芽, 茎尖剥离后培养至4~5个叶片, 用100 mg/L植物诱抗剂 AHO水剂喷施试管苗, 每隔2 d喷施一次, 共3次, 末次喷施2 d后取茎尖剥离培养, 获得再生试管苗?用电子显微镜负染色?ELISA?荧光定量RT-PCR检测再生试管苗的带病毒情况?结果显示, AHO对4个品种的马铃薯试管苗生长无影响, 用AHO处理后再茎尖剥离培养, 脱毒率均高于未处理的对照; 检测结果还显示AHO处理的马铃薯再生苗的带毒量也低于未处理的对照, 且随处理次数增加带毒量下降?研究结果表明, 利用植物诱抗剂AHO联合茎尖剥离培养方法可以提高脱除PVS?PVY的效率, 获得无病毒核心苗? 相似文献
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苹果潜隐病毒(Latent virus)研究——Ⅱ.苹果品种和矮生砧木潜隐病毒鉴定 总被引：9，自引：0，他引：9
在1980-1985年,对我国主要苹果产区的潜隐病毒种类进行调查鉴定,基本明确渤海湾果区、黄河故道果区和西北高原果区主栽苹果品种普遍潜带褪绿叶斑病毒、茎痘病毒和茎沟病毒,未检出其他病毒。营养系矮生砧木也普遍潜带这三种病毒。解决潜隐病毒为害的主要途径是培育无病毒母本树,栽培无病毒苗木。这方面的研究工作正在进行中。 相似文献
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我国是第一大苹果生产国。苹果茎沟病毒(apple stem grooving virus, ASGV)在我国各苹果产区分布广泛,侵染果树导致树势变弱,生长量减少,果实品质下降,严重威胁我国苹果产业的可持续发展。病毒检测是果树病毒病防控的重要前提。本研究根据苹果茎沟病毒外壳蛋白基因保守序列,设计了引物和探针,建立了微滴数字PCR (droplet digital PCR, ddPCR)检测方法。所建立的检测方法能特异性检出苹果茎沟病毒,检测重复性好,灵敏度比qRT-PCR法高10倍,可适用于田间样品及组培苗检测。该方法的建立为苹果茎沟病毒的检测提供了重要的技术手段。 相似文献
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我国北方部分苹果主产区病毒病的发生与检测 总被引：3，自引：0，他引：3
苹果病毒病在我国广泛发生,已成为限制我国苹果优质高产的关键因素。20世纪80年代曾对其做过详细的调查和研究,但近年来由于各地农业产业结构调整等因素,苹果病毒病的发生特点有了不同程度的变化。为了解目前我国苹果病毒病的发生情况,在我国北方苹果主产区山东、陕西、山西、辽宁、北京和黑龙江6个省市的部分地区采集苹果样品共计267份,经RT-PCR检测和扩增产物的克隆与测序分析表明在上述地区采集的样品中,苹果褪绿叶斑病毒(ACLSV)、苹果茎沟病毒(ASGV)、苹果茎痘病毒(ASPV)、苹果锈果类病毒(ASSVd)的发生率分别为66.7%~100.0%、38.1%~94.1%、4.8%~85.7%和4.8%~48.6%;苹果凹果类病毒(ADFVd)仅在山东的两个果园零星发生;6个省市样品中病毒复合侵染率分别为67.1%、92.1%、75.0%、88.2%、94.1%和76.2%。 相似文献
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苹果病毒病在世界各苹果产区广泛发生,危害严重,目前没有有效的防治药剂。苹果病毒具有潜隐危害、主要以嫁接方式传播的特点。危险性高的类病毒侵染导致苹果果实几乎失去经济价值。苹果病毒的检测是防止其传播和危害,繁育无病毒苗木和苹果树无毒化管理的一项关键技术。由于对病毒在苹果树体中的组织分布和浓度认知不到位,苹果病毒检测中存在样品采集和检测方法的误区。本文根据课题组十年来开展苹果病毒检测和研究工作的数据,简要概述了我国苹果病毒的发生危害特点,苹果病毒检测方法,详细给出了规范的样品采集、处理、总RNA提取、RT-PCR参数、检测引物、对照设置及检测结果判别的技术方法。 相似文献
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Hiroki Noda Noriko Yamagishi Hajime Yaegashi Fei Xing Jipeng Xie Shifang Li Tao Zhou Tsutae Ito Nobuyuki Yoshikawa 《Journal of General Plant Pathology》2017,83(2):83-90
The causal agent of apple mosaic disease has been previously thought to be solely caused by apple mosaic virus (ApMV). In this study, we report that a novel ilarvirus is also associated with apple mosaic disease. Next-generation sequencing analysis of an apple tree showing mosaic symptoms revealed that the tree was infected with three apple latent viruses (apple stem pitting virus, apple stem grooving virus, and apple chlorotic leaf spot virus) and a novel ilarvirus (given the name apple necrotic mosaic virus (ApNMV)) that is closely related to Prunus necrotic ringspot virus (PNRSV) and ApMV. The genome of ApNMV consists of RNA1 (3378 nt), RNA2 (2767 nt), and RNA3 (1956 nt). A phylogenetic analysis based on the coat protein amino acid sequences indicated that the novel virus belongs to the same subgroup 3 of the genus Ilarvirus as PNRSV and ApMV. The presence of mosaic leaves, which tend to be unevenly distributed in diseased apple trees, was correlated with the internal distribution of ApNMV. RT-PCR detection of mosaic-diseased apple trees in Japan indicated that ApNMV was detected in apple trees introduced from China, whereas ApMV was detected from cultivated apple trees in domestic orchards. Consistent with these findings, a survey of mosaic-diseased apple trees in major apple-producing provinces in China revealed that the majority of apple trees showing mosaic symptoms in China are infected with ApNMV. 相似文献
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Elimination of Candidatus phytoplasma phoenicium from two infected Lebanese varieties of almond by using different tissue culture techniques is reported. Except for the oxytetracycline therapy which totally inhibited the development of explants, stem cutting cultures associated with thermotherapy, shoot tip cultures associated or not with thermotherapy, and shoot tip micrografting were all suitable, either for shoot regeneration or for elimination of phytoplasma from the two varieties. However, stem cutting culture coupled with thermotherapy seemed to be the most effective for regeneration of phytoplasma-free plantlets. 相似文献
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L. Chen M.-R. Wang J.-W. Li C.-H. Feng Z.-H. Cui L. Zhao Q.-C. Wang 《Plant pathology》2019,68(5):997-1006
Production and maintenance of virus-free planting materials is pivotal for the control of viral diseases. The present study attempted to test exogenous application of melatonin for eradication of apple stem grooving virus (ASGV) from virus-infected in vitro shoots of apple cultivar Gala. Exogenous application of 15 μm melatonin to the shoot proliferation medium significantly increased the number of shoots and shoot length. The level of endogenous indole-3-acetic acid (IAA) was the highest in the shoots proliferating on the shoot proliferation medium containing 15 μm melatonin. Shoot regrowth levels were significantly higher in shoot tips of the virus-infected shoots cultured for 4 weeks on this medium than the control. In addition, culture of shoot tips of the virus-infected in vitro shoots proliferated for 4 weeks on this medium resulted in 95% of shoots being virus-free, while no virus-free shoots were obtained in shoot tips of the virus-infected shoots cultured without melatonin. Analyses by microtissue direct RT-PCR and RT-qPCR showed that ASGV concentration decreased in shoot tips of the virus-infected shoots proliferating on the medium containing 15 μm melatonin for 4 weeks. Virus localization showed that exogenous application of melatonin enlarged the virus-free area in the virus-infected shoot tips. These data provide explanations as to why exogenous application of melatonin can efficiently eradicate ASGV. Exogenous application of melatonin provides an alternative means for plant virus eradication and has the potential to produce virus-free plants. 相似文献
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Kazuya Nakamura Noriko Yamagishi Masamichi Isogai Sadao Komori Tsutae Ito Nobuyuki Yoshikawa 《Journal of General Plant Pathology》2011,77(1):48-53
To examine whether Apple latent spherical virus (ALSV) has spread among apple trees in an orchard, we surveyed 21 apple trees surrounding two ALSV-infected trees for virus
infection using a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). None of the 21 trees were infected,
indicating that ALSV has not spread from the infected trees to the neighboring apple trees since it was first detected in
1984. We analyzed seed embryos and seedlings derived from infected trees and detected ALSV in 10 of 223 seed embryos (4.5%)
and 10 of 227 seedlings (4.4%). From these results, we conclude that ALSV is seed-transmitted at a rate of ca. 4.5% in apple.
We also analyzed seed embryos and seedlings from uninfected apple trees that were hand-pollinated with pollen from infected
trees. We detected ALSV in only 1 of 260 seed embryos and in none of the 227 apple seedlings. This result indicated that the
seed transmission rate via infected pollen is only 0–0.38%. In situ hybridization analysis of ALSV-infected apple flower buds
showed that ALSV was present inside almost all pollen grains and in all ovary and ovule tissues, including the embryo sac
and inner integument. 相似文献
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Martin Verbeek Paul van Dijk Peter M. A. van Well 《European journal of plant pathology / European Foundation for Plant Pathology》1995,101(3):231-239
Mechanical inoculation tests and ELISA with sap from garlic plants used for sanitation by meristem-tip culture revealed four viruses, viz. garlic common latent virus (GCLV) (carlavirus), the garlic strains of leek yellow stripe virus (LYSV-G), onion yellow dwarf virus (OYDV-G) (aphid-borne potyviruses), and onion mite-borne latent virus (OMbLV-G) (taxonomically unassigned virus). The same tests performed on explants grownin vitro showed elimination efficiencies of 100% for LYSV-G, 92% for OYDV-G, 62% for GCLV, and less then 54% for OMbLV-G.Meristem tips excised from garlic cloves and bulbils, 0.15–1.0 mm in size, were tested for regeneration and efficiency of virus elimination after transfer to Murashige and Skoog medium. Successful regeneration into plantlets was obtained with 71% of the meristems from cloves and 72% of those from bulbils, but virus elimination was easiest from cloves: 38% of all explants from cloves and 25% of those from bulbils were virus-free. The efficiency of elimination increased with increasing weight of the cloves, irrespective of the virus. Small tip size seemed to favour virus elimination, but sizes smaller than 0.4 mm led to increasing failure of regeneration.Micropropagation was most successful when cytokinins were omitted from the medium and the garlic shoot was split. Multiplication factors of 3–6 were obtained. 相似文献