表达黄瓜花叶病毒卫星RNA的转基因烟草耐烟草花叶病毒

表达黄瓜花叶病毒(CMV)的卫星RNA的转基因烟草可以抗CMV的侵染.为了决定这些植物对其它病毒,如烟草花叶病毒(CMV)的安全性,我们对这些植物接种CMV.结果发现 卫星RNA可减弱TMV引起的症状,井降低病情指数,然而却对TMV在植物中的积累没有明显影响.这一发现有助于对卫星RNA抗病机制的理解,有助于含卫星RNA的生防制剂及表达病毒卫星RNA的转基因植物的应用.本文在国际上首次报道卫星RNA可减弱非相关病毒引起的症状.1 材料和方法1.1 病毒及植物烟草花叶病毒(TMV),烟草G140种子及枯班三生烟种子均为中国科学院微生物研究所病毒室保存;含黄瓜花叶病毒卫星RNA-R1基因的烟草G140(烟Sat-G140)为中国科学院微生物研究所病毒室培育,所用材料为第三代种子.     

表达黄瓜花叶病毒卫星RNA的转基因烟草耐烟草花叶病毒

表达黄瓜花叶病毒(CMV)的卫星RNA的转基因烟草可以抗CMV的侵染.为了决定这些植物对其它病毒,如烟草花叶病毒(CMV)的安全性,我们对这些植物接种CMV.结果发现 卫星RNA可减弱TMV引起的症状,井降低病情指数,然而却对TMV在植物中的积累没有明显影响.这一发现有助于对卫星RNA抗病机制的理解,有助于含卫星RNA的生防制剂及表达病毒卫星RNA的转基因植物的应用.本文在国际上首次报道卫星RNA可减弱非相关病毒引起的症状.1 材料和方法1.1 病毒及植物烟草花叶病毒(TMV),烟草G140种子及枯班三生烟种子均为中国科学院微生物研究所病毒室保存;含黄瓜花叶病毒卫星RNA-R1基因的烟草G140(烟Sat-G140)为中国科学院微生物研究所病毒室培育,所用材料为第三代种子.     

从烟草上分离的黄瓜花叶病毒

黄瓜花叶病毒(CMV)危害烟草已成为烟区严重病害之一,如山东烟区CMV引起的烟草病毒病发病率达50%以上。CMV在福建烟区的发病率也达15～30%。上述调查系根据烟草田间症状表现以及温室内不同鉴别寄主的反应和抗性测定得到。为了进一步     

从烟草上分离的黄瓜花叶病毒

黄瓜花叶病毒(CMV)危害烟草已成为烟区严重病害之一,如山东烟区CMV 引起的烟草病毒病发病率达50%以上。GMV 在福建烟区的发病率也达15～30%。上述调查系根据烟草田间症状表现以及温室内不同鉴别寄主的反应和抗性测定得到。为了进一步研究与肯定CMV 引起的烟草病毒病,有必要由病株分离病毒和进行电子显微镜观察鉴定。     

Fuzzy相关综合评判模型在辣椒品种抗病毒病鉴定中的应用

本文以辣椒苗期对烟草花叶病毒(TMV)和黄瓜花叶病毒(CMV)的抗性作评判因子,应用Fuzzy相关综合评判模型,对品种田间抗病毒性表现进行评判。结果表明:与田间抗病毒性表现符合率为92.31%,说明该方法具有较高的可靠性,可作为其他作物抗病性鉴定的借鉴。     

Fuzzy相关综合评判模型在辣椒品种抗病毒病鉴定中的应用

本文以辣椒苗期对烟草花叶病毒(TMV)和黄瓜花叶病毒(CMV)的抗性作评判因子,应用Fuzzy相关综合评判模型,对品种田间抗病毒性表现进行评判。结果表明:与田间抗病毒性表现符合率为92.31%,说明该方法具有较高的可靠性,可作为其他作物抗病性鉴定的借鉴。     

保存CMV毒源的试管烟苗方法

黄瓜花叶病毒（简称CMV）有极其广泛的天然寄主，能侵染75科近700种植物，迄今还缺少有效的防治方法，有植物界的“流感’与“癌症”之称。烟草受害尤重，一股流行年份烟草产值下降25～40％，严重的达50％以上乃至绝收。为此众多生物技术工作者分别从不同角度开展了CMV的研究。     

延边地区烟草病毒种类及病毒病的防治

对田间烟草进行病毒病种类的调查和鉴定结果表明,延边地区感染烟草的病毒主要是烟草花叶病毒(TMV)和黄瓜花叶病毒(cMV),而以两种病毒的混合侵染为主,达80-1 00％。弱毒疫苗N14．和CMV卫星RNA生防制剂S5,混合使用,或CMV卫星RNA生防制剂S512单独使用防治烟草病毒病,与不使用的对照比较,防治效果明显,达70.2—77.1％,上等烟比率达22.3—30.9％,而对照只有9．2-1 9.2％,亩收入比对照增加21．7—8 0.8％。     

浙江油菜花叶病毒类型研究

浙江各地引起油菜花叶病的病毒类型有芜菁花叶病毒(TpMV)、黄瓜花叶病毒(CMV)、菸花叶病毒(TMV)及芜菁花叶病毒和黄瓜花叶病毒的复合病(TpMV+cMV)。芜菁花叶病毒有TpMV98。和TpMV98 两个毒株,主要区别： TpMV98不侵染心叶菸,TpMV58 侵染心叶菸引起的系统性枯斑和系统性环斑能随环境的影响相互转变。黄瓜花叶病毒有CMV69和CMV72 两个毒株,主要区别：CMV69,回接不上本地油菜,CMV72 能回接上本地油菜。菸花叶病毒毒株(TMV45)侵染多种十字花科植物,在胜利油菜上引起系统花叶,在苋桥红萝卜上是无病状带毒,在农特400号菸上引起局部枯斑。TPMV98和CMV69 或CMV72 形成复合病,可以汁液接种分离,再混合再分离。根据采集样品按类型和毒株来看,浙江大田油菜花叶病,绝大多数是芜菁花叶病毒类型中的TpMV98毒株,复合病和黄瓜花叶病毒分布也相当广。辣根(Cochlearia armoracia L.)上所采的四个样品均属芜菩花叶病毒中侵染心叶菸的TpMV58毒株,辣根的宿根可以保存病毒,可能是这一毒株的越夏寄主之一。     

重庆烟草地区ELISA检测

为调查重庆主要烟草地区的病毒病种类,采用没脸免疫吸附法(ELISA)对采自重庆涪陵、彭水、万州、黔江、武隆、酉阳的258份的烟草样品进行了病毒病种类的检测.检测结果表明,其中274份样品呈阳性。主要受到7种病毒病的侵染,主要包括:烟草花叶病毒(Tobacco mosaic virus,TMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、马铃薯Y病毒(Potato virus Y,PVY)、马铃薯X病毒(Potato virus X,PVX)、芜菁花叶病毒(Turnip mosaic virus,TuMV)、蚕豆萎蔫病毒2号Broad bean wilt virus 2,BBWV-2)、烟草蚀纹病毒(Tobacco etch virus,TEV)。其中TMV的检出率最高,达50%,已成为重庆地区烟草优势病原。     

Co-electroporation of rice protoplasts with RNAs of cucumber mosaic and tobacco mosaic viruses

Cucumber mosaic virus (CMV) RNA was used to study electroporation conditions suitable for protoplasts from rice suspension cultures. Rice protoplasts required a stronger and shorter electric pulse than tobacco protoplasts for introduction of viral RNA. Under optimized conditions, CMV infection was established in 65 % of electroporated protoplasts. In contrast, electroporation with tobacco mosaic virus (TMV) RNA did not result in infection of rice protoplasts. However, when TMV RNA was electroporated into rice protoplasts together with CMV RNA, TMV production was demonstrated in 15 % of protoplasts. Differential staining with fluorescent antibodies against the two viruses showed that the protoplasts producing TMV were without exception also infected by CMV. The results show that CMV replicates in rice protoplasts by itself, whereas TMV does so only with the aid of CMV.Abbreviations CMV cucumber mosaiv virus - PBS phosphate buffered saline - TMV tobacco mosaic virus.     

Salicylic acid-induced resistance to Cucumber mosaic virus in squash and Arabidopsis thaliana: contrasting mechanisms of induction and antiviral action

Salicylic acid (SA)-induced resistance to Cucumber mosaic virus (CMV) in tobacco (Nicotiana tabacum) results from inhibition of systemic virus movement and is induced via a signal transduction pathway that also can be triggered by antimycin A, an inducer of the mitochondrial enzyme alternative oxidase (AOX). In Arabidopsis thaliana, inhibition of CMV systemic movement also is induced by SA and antimycin A. These results indicate that the mechanisms underlying induced resistance to CMV in tobacco and A. thaliana are very similar. In contrast to the situation in tobacco and A. thaliana, in squash (Cucurbita pepo), SA-induced resistance to CMV results from inhibited virus accumulation in directly inoculated tissue, most likely through inhibition of cell-to-cell movement. Furthermore, neither of the AOX inducers antimycin A or KCN induced resistance to CMV in squash. Additionally, AOX inhibitors that compromise SA-induced resistance to CMV in tobacco did not inhibit SA-induced resistance to the virus in squash. The results show that different host species may use significantly different approaches to resist infection by the same virus. These findings also imply that caution is required when attempting to apply findings on plant-virus interactions from model systems to a wider range of host species.     

不同CMV分离物侵染寄主的超微结构变化

应用电镜观察了黄瓜花叶病毒ＣＭＶ不同分离物侵染寄主的细胞超微结构变化。来自一患红（Ｓａｌｖｉａｓｐｌｅｎｄｅｎｓ）的不含卫星ＲＮＡ分离物Ｍ－２２侵染心叶烟,病毒粒子散布于细胞质,在液泡中形成大片病毒粒子结果,液泡膜边缘产生小泡结构,完整的病毒粒子穿过胞间连丝在细胞间运转,胞间连丝中央部分有扩张现象。     

应用RNAi技术培育抗2种病毒病的转基因烟草

分别提取烟草普通花叶病(TMV)和烟草黄瓜花叶病（CMV）的病毒RNA。经反转录和外壳蛋白阅读框PCR扩增,获得TMV和CMV外壳蛋白基因cDNA, 分别进行两种病毒已知株系cDNA序列比对获得各自的保守序列,设计干涉序列,将干涉片段扩增产物连接到pMD18-T的相邻酶切位点,制备融合序列,并将其正向和反向序列插入pUCCRNAi载体,再转化到pCAMBIA2300-35S-OCS表达载体中。利用农杆菌LBA4404侵染烟草K326,获得3份含有TMV和CMV外壳蛋白基因干涉序列的转化材料,经分子鉴定证实干涉序列已导入烟草,并采用荧光定量PCR技术对其mRNA表达差异进行分析。抗病性调查表明转化烟株对TMV和CMV抗性都显著增强。     

安徽烟草病毒类型及优势种初步研究

1978年,烟草病毒病在安徽烟区流行,导致凤阳县烟叶总产损失92.3％,引起了普遍震惊。1981—1984年作者对来自16个县、市552个病毒材料,经生物测定、血清反应、电镜观察,初步分离出CMV、TMV、PVY和PVX四种病毒。它们分别约占检测总数的82.79％、4.53％、2.54％和0.36％,其中CMV与长期视为优势种的TMV比值为18.3,除此,尚有约占检测总数9.8％的CMV和TMV、CMV和PVY复合侵染,以及不明类型的毒株。通过对田间烟草以及其他植物花叶病株的实际检测,进一步表明:CMV在烟区分布范围极广、出现频次最多,已形成了复杂的循环侵染系统,成为近期内烟草病毒病持续流行危害的首要毒原。     

Interaction of cucumber mosaic virus and potato virus y with tobacco mosaic virus

A study was performed on the interaction of cucumber mosaic virus (CMV) of potato virus Y (PVY) with tobacco mosaic virus (TMV). Interference was evaluated using tobacco plantsNicotiana tabacum cv. Java responding to CMV and PVY with a systemic infection and to TMV with local necrotic lesions. The decrease in TMV — induced lesion number gave evidence of a decrease in susceptibility caused by the previous infection with CMV or PVY, the decrease of lesion enlargement demonstrated a decreased TMV reproduction in the plants previously infected with CMV or PVY. The interference concerned was incomplete, as evaluated from reproduction of the challenging TMV and from the decrease in susceptibility of the host to TMV brought about by the first infection with CMV or PVY.     

Virus Movement Protein Gene Mediated Resistance Against Cucumber Mosaic Virus Infection

Tobacco (Nicotiana tabactum L. ) "NC89" plants were transformed with deletion mutant of cucumber mosaic virus (CMV) movement protein (MP) gene and full-length CMV MP gene, respectively. The transformed plants were analyzed with polymerase chain reaction (PCR), PCR-Southem, Southern and Western blots. R0 generation of the transgenic plants were inoculated with CMV. Five out of 10 lines of tobacco plants (BMPK) transformed with CMV MP deletion mutant gene showed high resistance to CMV infection and remained symptomless for up to 50 days post-inoculation. In contrast, tobacco plants (BMPR) transformed with full-length CMV MP gene did not show resistance to CMV infection. However, most of the infected full-length CMV MP gene transgenic plants recovered by showing none or very mild mosaic symptoms in 40 days post-inoculation. The results of R1 generation of the BMPK transgenic plants tested under field conditions showed that all 5 lines of transgenic plants could delay the virus disease development.     

Cucumber mosaic virus movement protein affects sugar metabolism and transport in tobacco and melon plants

In addition to its influence on plasmodesmal function, tobacco mosaic virus movement protein (TMV‐MP) causes an alteration in carbon metabolism in source leaves and in resource partitioning among the various plant organs. The present study was aimed at characterizing the influence of cucumber mosaic virus (CMV)‐MP on carbohydrate metabolism and transport in both tobacco and melon plants. Transgenic tobacco plants expressing the CMV‐MP had reduced levels of soluble sugars and starch in their source leaves and a significantly reduced root‐to‐shoot ratio in comparison with control plants. A novel virus‐vector system was employed to express the CMV‐coat protein (CP), the CMV‐MP or the TMV‐MP in melon plants. This set of experiments indicated that the viral MPs cause a significant elevation in the proportion of sucrose in the phloem sap collected from petioles of source leaves, whereas this sugar was at very low levels or even absent from the sap of control melon plants. The mode by which the CMV‐MP exerts its effect on phloem‐sap sugar composition is discussed in terms of possible alterations in the mechanism of phloem loading.