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植物抗病毒道传工程中利用病毒衣壳蛋白基因、弱毒株系的完整基因组、病毒反义RNA序列和RNA随体序列等方法获得作物抗病毒转基因植物,本文列举了这些方法的最新应用实例。 相似文献
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水稻矮缩病毒(Rice dwarf virus,RDV)为双层壳、二十面体的双链RNA病毒,是水稻的重要病原物之一.文中比较了RDV中国福建分离物与日本分离物基因组各片段核苷酸序列的相似性,发现RDV两个不同分离物基因组相应片段的相似性均在92%以上,最高可达96%;分析了相应基因组片段编码的结构蛋白P1、P2、P3、P5、P7、P8及以前一直被认为是非结构蛋白的P9和非结构蛋白Pns4、Pns6、Pns10 、Pns11和Pns12在RDV复制、组装过程中的功能;回顾了RDV粒子三维结构的研究概况,对RDV粒子内、外层衣壳的晶体学结构作了较详细的描述;简要介绍了RDV复制与组装的机制,发现核心蛋白与dsRNA间相互作用成为一个单元是病毒RNA的分拣及包装到病毒粒子核心内的可能机制.同时,指出基于PDR的植物抗病毒基因工程在水稻矮缩病毒防治上的可能性. 相似文献
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利用RNA介导的抗病性获得抗2种病毒的转基因烟草 总被引：11，自引：0，他引：11
RNA介导的病毒抗性(RMVR)是近年发展起来的一种新的植物抗病毒基因工程策略,具有抗病性强、抗性持久、生物安全性高等特点。利用该策略培育多抗病毒植株具有广阔的应用前景和重要的实践意义。本研究将非翻译的马铃薯X病毒的衣壳蛋白(PVX-CP)基因和非翻译的马铃薯Y病毒的衣壳蛋白(PVY-CP)基因组成嵌合基因,构建植物表达载体pROKXY,利用农杆菌介导法转化烟草NC89,获得6株对PVX和PVY的混合侵染表现为免疫的转基因植株。分子分析表明,这种抗性为RNA介导的病毒抗性。这一研究结果为利用RMVR进行植物多抗病毒育种提供了重要数据和资料。 相似文献
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棉铃虫核型多角体病毒分子生物学和基因工程研究进展 总被引：1，自引：1，他引：1
棉铃虫核型多角体病毒(HaSNPV)是棉铃虫专一性病原物,隶属于杆状病毒科核型多角体病毒属.对其分子生物学和基因工程的研究主要包括以下几个方面:测定了HaSNPV G4株和C1株基因组核苷酸全序列,并与其它病毒进行了同源性比较;研究了HaSNPV部分基因的结构、转录、表达及其功能.构建了HaSNPV Bac to Bac杆状病毒表达系统;重组病毒杀虫剂的研究为HaSNPV大面积防治棉铃虫展示了广阔的前景.随着棉铃虫核型多角体病毒分子生物学和基因工程研究的不断深入,重组病毒杀虫剂将在棉铃虫综合防治中发挥更为重要的作用. 相似文献
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水稻黑条矮缩病毒(Rice black-streaked dwarf virus, RBSDV)基因组含有5′帽子,无3′poly(A)尾巴,且不同基因组片段的5′和3′末端序列均十分保守。本研究以RBSDV S3和S10为对象,分析其非编码区对翻译的调控作用。研究发现S3和S10的5′UTR在有无帽子时均可以正向调控报告基因Fluc的翻译,具备核糖体内部进入位点(IRES)活性,且5′末端的基因组保守序列及邻近序列与IRES活性密切相关;而对应的3′UTR则抑制5′UTR对翻译的正调控效应,其分子基础是5′UTR和3′UTR之间的RNA-RNA互作,且该互作也抑制帽子依赖型翻译的效率。这是首次关于有帽子植物RNA病毒中IRES元件的报道,研究结果对了解植物双链RNA病毒基因组非编码区对翻译的调控作用具有重要科学意义。 相似文献
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番茄灼烧病毒属——番茄上新发现的一类植物类小RNA病毒 总被引：1，自引：0，他引：1
番茄灼烧病毒(170mato torrado virus,ToTV)、番茄顶点坏死病毒(Tomato apex necrosis virus,TOANV)和蕃茄枯萎病毒(Tomato marchitez virus,ToMarV)是近几年来在番茄上发现的一类植物类小RNA病毒,分别引起番茄"灼烧病"(torrado disense)和"枯萎病"(marchitez disease).该类病毒粒体形态是一种直径约为28 nm的等轴多面体病毒.其基因组是由两个长度分别约为7 kb(RNA1)和5 kb(RNA2)的ssRNA分子组成.ICTV植物类小RNA病毒研究小组最近把它们归为一个新的植物病毒属,即番茄灼烧病毒属(Torradovirus).本文就这类病毒的生物学、分子生物学特征分别作简单介绍. 相似文献
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RNA沉默是植物中一种保守的抗病毒机制,其以大量病毒来源的小干扰RNA(virus-derived small interfering RNAs,vsiRNAs)的产生为标志,病毒在侵染寄主过程中可通过vsiRNAs靶向寄主转录本以对抗这种防御机制。BYDV-GAV引起的小麦黄矮病导致小麦黄化和矮化症状,对小麦生产构成严重威胁。通过深度测序技术,分析了感染BYDV-GAV的感病小麦品种‘小偃6号’中vsiRNAs的特征,共得到11 384个vsiRNAs,并预测到37 784个寄主靶基因。发现来源于BYDV-GAV基因组的正义和反义链的vsiRNAs的数量分布大致相等,在5’末端具有A和C偏好性,长度主要在21nt~22nt。靶基因的功能分析表明这些靶基因参与了广泛的生物学功能,尤其是在寄主-病原物互作中占的比重最大。选取25个参与寄主-病原互作的抗性相关基因进行定量验证,发现接种BYDV-GAV后有15个明显下调,6个上调,4个微弱下调,表明测序结果和靶基因预测可靠。推测BYDV-GAV可以通过vsiRNAs干扰寄主抗性基因表达和信号转导,从而实现对感病寄主的侵染。研究结果对揭示BYDV-GAV与小麦互作的分子机制具有重要意义。 相似文献
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RNA沉默是植物中一种保守的抗病毒机制,其以大量病毒来源的小干扰RNA(virus-derived small interfering RNAs,vsiRNAs)的产生为标志,病毒在侵染寄主过程中可通过vsiRNAs靶向寄主转录本以对抗这种防御机制。BYDV-GAV引起的小麦黄矮病导致小麦黄化和矮化症状,对小麦生产构成严重威胁。通过深度测序技术,分析了感染BYDV-GAV的感病小麦品种‘小偃6号’中vsiRNAs的特征,共得到11 384个vsiRNAs,并预测到37 784个寄主靶基因。发现来源于BYDV-GAV基因组的正义和反义链的vsiRNAs的数量分布大致相等,在5’末端具有A和C偏好性,长度主要在21nt~22nt。靶基因的功能分析表明这些靶基因参与了广泛的生物学功能,尤其是在寄主-病原物互作中占的比重最大。选取25个参与寄主-病原互作的抗性相关基因进行定量验证,发现接种BYDV-GAV后有15个明显下调,6个上调,4个微弱下调,表明测序结果和靶基因预测可靠。推测BYDV-GAV可以通过vsiRNAs干扰寄主抗性基因表达和信号转导,从而实现对感病寄主的侵染。研究结果对揭示BYDV-GAV与小麦互作的分子机制具有重要意义。 相似文献
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ABSTRACT Ribosome-inactivating proteins (RIPs) are N-glycosidases that remove specific purine residues from the sarcin/ricin (S/R) loop of the large rRNA and arrest protein synthesis at the translocation step. In addition to their enzymatic activity, RIPs have been reputed to be potent antiviral agents against many plant, animal, and human viruses. We recently showed that pokeweed antiviral protein (PAP), an RIP from pokeweed, inhibits translation in cell extracts by binding to the cap structure of eukaryotic mRNA and viral RNAs and depurinating these RNAs at multiple sites downstream of the cap structure. In this study, we examined the activity of three different RIPs against capped and uncapped viral RNAs. PAP, Mirabilis expansa RIP (ME1), and the Saponaria officinalis RIP (saporin) depurinated the capped Tobacco mosaic virus and Brome mosaic virus RNAs, but did not depurinate the uncapped luciferase RNA, indicating that other type I RIPs besides PAP can distinguish between capped and uncapped RNAs. We did not detect depurination of Alfalfa mosaic virus (AMV) RNAs at multiple sites by PAP or ME1. Because AMV RNAs are capped, these results indicate that recognition of the cap structure alone is not sufficient for depurination of the RNA at multiple sites throughout its sequence. Furthermore, PAP did not cause detectable depurination of uncapped RNAs from Tomato bushy stunt virus (TBSV), Satellite panicum mosaic virus (SPMV), and uncapped RNA containing poliovirus internal ribosome entry site (IRES). However, in vitro translation experiments showed that PAP inhibited translation of AMV, TBSV, SPMV RNAs, and poliovirus IRES dependent translation. These results demonstrate that PAP does not depurinate every capped RNA and that PAP can inhibit translation of uncapped viral RNAs in vitro without causing detectable depurination at multiple sites. Thus, the cap structure is not the only determinant for inhibition of translation by PAP. 相似文献
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ABSTRACT A new variety of Nicotiana, N. edwardsonii var. Columbia, was evaluated for its capacity to serve as a new source for virus resistance genes. Columbia was developed from a hybridization between N. glutinosa and N. clevelandii, the same parents used for the formation of the original N. edwardsonii. However, in contrast to the original N. edwardsonii, crosses between Columbia and either of its parents are fertile. Thus, the inheritance of virus resistance genes present in N. glutinosa could be characterized by using Columbia as a bridge plant in crosses with the susceptible parent, N. clevelandii. To determine how virus resistance genes would segregate in interspecific crosses between Columbia and N. clevelandii, we followed the fate of the N gene, a single dominant gene that specifies resistance to Tobacco mosaic virus (TMV). Our genetic evidence indicated that the entire chromosome containing the N gene was introgressed into N. clevelandii to create an addition line, designated N. clevelandii line 19. Although line 19 was homozygous for resistance to TMV, it remained susceptible to Tomato bushy stunt virus (TBSV) and Cauliflower mosaic virus (CaMV) strain W260, indicating that resistance to these viruses must reside on other N. glutinosa chromosomes. We also developed a second addition line, N. clevelandii line 36, which was homozygous for resistance to TBSV. Line 36 was susceptible to TMV and CaMV strain W260, but was resistant to other tombusviruses, including Cucumber necrosis virus, Cymbidium ringspot virus, Lettuce necrotic stunt virus, and Carnation Italian ringspot virus. 相似文献
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The effect of different nutritional amendments on the growth of tomato seedlings subjected to combined infection byFusarium oxysporum f.sp.lycopersici (Fol) and either Tomato Bushy Stunt Virus (TBSV) or Tomato Mosaic Virus (ToMV) was investigated. The growth of healthy or infected tomato seedlings was repressed more when they were maintained under conditions of nitrogen deficiency than in nutrient solutions devoid of potassium or phosphorus. Plants infected with the fungus and either virus had lower fresh weights and smaller leaf areas than those infected by any of these pathogens alone. With the exception of seedlings grown in potassium-deficient solutions, co-infection resulted in no fundamental differences in virus concentration per plant. The concentration of both TBSV and ToMV was higher per unit fresh weight of leaf in co-infected plants maintained in all types of nutrient. It is suggested that the presence of the fungus in the plant tissues may be a more important factor than nutrition in promoting virus replication. 相似文献
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B. D. Harrison 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):1-12
Knowledge of the nucleotide sequences in the genomic nucleic acid of several potato viruses has enabled the open reading frames to be identified. These open reading frames are expressed by a variety of strategies, to produce proteins with functions in virus nucleic acid replication, virus particle production, cell-to-cell transport of virus and virus transmission by vectors. The activity of such proteins depends on their interactions with other viral or non-viral materials.Several other biological properties of plant viruses can also be related to individual viral gene products. For example, in plants co-infected with a specific pair of unrelated viruses, one virus can benefit from an ability to use the gene product of the second virus in replication, cell-to-cell transport or transmission by vectors. Similarly, different host resistance genes are targeted against viral replicase, movement protein or coat protein. Thus it is becoming possible to relate gene-for-gene (or more accurately, viral gene domain-host gene) interactions to events at the molecular level. Genetically engineered resistance to plant viruses likewise can be targeted against individual viral genes, and probably also against viral regulatory sequences. Such transgenic resistance seems likely to be as durable as conventional host resistance but durability should be improved by producing plants with combinations of resistances of different kinds, either conventional or genetically engineered, or both. 相似文献
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两重RT-PCR同步检测菊花B病毒和番茄不孕病毒 总被引：2，自引：0，他引：2
根据已报道的菊花B病毒（CVB）和番茄不孕病毒（TAV）外壳蛋白基因序列合成两条寡核苷酸引物,用RT-PCR的方法对这两种病毒的外壳蛋白基因进行了扩增,得到与预期大小相符的特异扩增条带。将其克隆到pGEM T中,经序列分析表明所克隆的是CVB和TAV的CP基因,与已知的序列相比,CVB的同源性分别为82%、85％,而TAV的同源性为98%。利用两重RT-PCR成功同步检测这两种病毒,从而建立了一种能同时检测两种病毒的快速、简便、灵敏的分子检测方法。 相似文献
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扬州蚕豆病毒病的鉴定及野豌豆潜隐病毒M扬州分离物基因组克隆及其序列分析 总被引：1，自引：1，他引：0
对田间疑似病毒侵染的蚕豆样品,提取总RNA,分离siRNA,建库并通过高通量测序,其结果经velvet拼接组装, 经Blastn与NCBI比对分析,发现该样品感染4种病毒:野豌豆潜隐病毒M (vicia cryptic virus M,VCV-M)、野豌豆潜隐病毒 (vicia cryptic virus, VCV)、紫云英矮宿病毒 (milk vetch dwarf virus, MDV) 和三叶草黄脉病毒 (clover yellow vein virus, ClYVV)。其中,有14条VCV-M相关的contigs。RT-PCR扩增并拼接获得VCV-M-YZ基因组,其长度为3 434 nt, 其5′-UTR和3′-UTR 分别为142 nt和117 nt,各自形成与该属代表病毒南方番茄病毒 (southern tomato virus, STV) 5′-UTR和3′-UTR相似的高级结构,且A+U含量均较高。 VCV-M-YZ与已报道的VCV-M基因组序列一致性在98%以上。系统进化分析表明,VCV-M-YZ与已报道的VCV-M属于一个种,无明显的地理分布差异。本研究结果丰富了VCV-M群体的基因组遗传信息,为VCV-M的监测及深入研究提供了基础。 相似文献
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Minimal Set of Metabolic Pathways Suggested from the Genome of Onion Yellows Phytoplasma 总被引：1，自引：0，他引：1
Kenro OSHIMA Shin-ichi MIYATA Toshimi SAWAYANAGI Shigeyuki KAKIZAWA Hisashi NISHIGAWA Hee-Young JUNG Ken-ichiro FURUKI Masaki YANAZAKI Shiho SUZUKI Wei WEI Tsutomu KUBOYAMA Masashi UGAKI Shigetou NAMBA 《Journal of General Plant Pathology》2002,68(3):225-236
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R. N. Campbell S. T. Sim H. Lecoq 《European journal of plant pathology / European Foundation for Plant Pathology》1995,101(3):273-282
Zoospores of 12 isolatesO. bornovanus from geographically diverse sites and representing the three host specific cucurbit strains were tested as vectors for seven viruses using watermelon bait plants and the in vitro acquisition method. All isolates of the cucumber, melon, and squash strains transmitted melon necrotic spot carmovirus (MNSV) and cucumber necrosis tombusvirus (CNV) but none transmitted petunia asteroid mosaic tombusvirus (PAMV) or tobacco necrosis necrovirus (TNV). The isolates varied as vectors of three other carmoviruses: cucumber leaf spot virus (CLSV); cucumber soil borne virus (CSBV); and squash necrosis virus (SqNV). All cucumber isolates transmitted CLSV and SqNV but not CSBV. Some of the melon isolates transmitted CLSV and SqNV but none transmitted CSBV. Two squash isolates transmitted CSBV and SqNV but not CLSV. Two isolates ofO. brassicae transmitted only TNV and a third did not transmit any of the viruses. The species of bait plant sometimes affected transmission. The most efficient vector strains ofO. bornovanus, as determined by reducing zoospores and virus in the inoculum, were the cucumber strain for CLSV; the cucumber strain for CNV if cucumber was the bait plant or melon strain if watermelon was the bait plant; and the squash strain for SqNV. The plurivorous strain ofO. brassicae was the most efficient vector of TNV.Olpidium bornovanus is the first vector reported for CSBV and is confirmed as a vector of SqNV. It is proposed that all carmoviruses may have fungal vectors.Ligniera sp. did not transmit any of the viruses in one attempt.Abbreviations CLSV
cucumber leaf spot virus
- CNV
cucumber necrosis virus
- CSBV
cucumber soil borne virus
- MNSV
melon necrotic spot virus
- PAMV
petunia asteroid mosaic virus
- SqNV
squash necrosis virus
- TNV
tobacco necrosis virus
- TBSV
tomato bushy stunt virus 相似文献