新疆西瓜花叶病毒2号外壳蛋白的研究

 本文以马铃薯Y病毒组（potyvirus group）确定成员西瓜花叶病毒2号（WMV-2）为研究对象，对新疆和澳大利亚WMV-2的外壳蛋白制备、快速蛋白液相色谱（FPLC）肽图谱进行了研究和比较。通过分析两者外壳蛋白同源性的大小，证明新疆和澳大利亚WMV-2是同种马铃薯Y病毒的不同株系。从而说明外壳蛋白的FPLC肽图谱可成功地用于马铃薯Y病毒组的鉴定和分类。     

Production of monoclonal antibodies against synthetic peptides of the N-terminal region of Potato virus Y coat protein and their use in PVY strain differentiation

Antibodies were prepared against two synthetic peptides, P19 and P11, derived from the coat protein N-terminal region of two pepper isolates of Potato virus Y from Tunisia (PVY-P21 and PVY-P2, respectively). The peptides were selected by comparing the predicted amino acid sequences of three pepper and four potato PVY isolates on the basis of their polymorphism and hydrophilicity. Sera with high titres were only obtained against P19. Three MAbs, raised in response to P19, reacted with the homologous virus (PVY-P21) in TAS-ELISA. When tested against a broad range of PVY isolates and related viruses, MAb 3C5 proved to be PVY species specific, whereas MAbs 8A4 and 1D6 reacted specifically with standard isolates of PVY^O, PVY^C and PVY^N-W strains, but not with other PVY isolates. Consequently, epitope(s) recognized by 8A4 and 1D6 MAbs may be specific to a PVY group comprising all serologically PVY^non–N isolates. Surprisingly, and unlike isolate PVY-P21, many Tunisian field pepper isolates did not carry this epitope(s), thus revealing serological heterogeneity within the PVY pepper group. As PVY is one of the most economically important plant pathogens in a range of crops, including pepper, these MAbs will provide a useful tool for practical diagnosis and strain identification of PVY.     

Weed Flora of Andhra Pradesh

Abstract

A large scale white grub control campaign was launched by the Marathwada Agricultural University, Parbhani. Pit trapping indicated that there was an initial beetle population of about 50 million in an area of 876 ha. About 9,000,000 beetles were collected mechanically and destroyed. One spraying of 12,000 babul and neem trees with 0.4% DDT killed 88% of the beetles. The percentage reduction in the plant population of rice, chillies, jowar and sugarcane, due to white grub attack was 1.83, 6.34, 3.59 and 3.02 in the treated area as against 65.54, 56.06, 49.68 and 8.94 in the untreated area. In cotton there was no reduction in plant population in the treated area. There was an increase in yield in paddy, chilli, cotton and jowar to the extent of 164.5, 102.2, 27.8 and 12.7% respectively in the treated area of Ratoli. There was an 80% reduction in the white grub larval population.     

Resistance to Turnip mosaic virus in Brassica rapa and B. napus and the analysis of genetic inheritance in selected lines

Forty-two Brassica rapa and Brassica napus lines were tested for resistance to Turnip mosaic virus (TuMV) isolates representing the three major pathotypes in Europe. Of these lines, 11 were susceptible to all pathotypes; nine were resistant to one pathotype; eight were resistant to two pathotypes; and 14 were resistant to all three pathotypes. Of the lines tested, 23 were either able to, or had the potential to, discriminate between two different pathotype-3 isolates. Genetic models for inheritance of resistance were proposed for four B. rapa lines: Jong Bai No. 2 had dominant resistance to pathotype 1 conferred by a single allele; PI418957C and Jin G 55 had recessive resistance to pathotype 4 where a single allele was required; PI418957C also had recessive resistance to pathotype 3 where a model with one of two epistatic, unlinked loci was proposed. Jong Bai No. 1 also had recessive resistance to pathotype 3, apparently conferred by alleles at three loci, where any two of the three loci were epistatic and required for resistance.     

Control of virus diseases in intensively cultivated vanilla plots of French Polynesia

The results of long term virus surveys in intensively cultivated vanilla plots in the Society Islands (French Polynesia), between 1999 and 2007 are reported here. The data confirmed a potential for high incidence of aphid borne viruses in particular Cucumber mosaic virus (CMV) and Watermelon mosaic virus (WMV) as well as the non vectored Cymbidium mosaic virus (CymMV). CMV had a particularly high prevalence (over 30% of the plots) and could severely damage up to 50% of the vines before blossom. Severe outbreaks of CMV were correlated to the presence of the weed Commelina diffusa as a reservoir of virus and aphid vectors. The application in 2003 of simple prophylactic measures resulted in a sharp reduction of virus incidence in new plantations, compared to levels of virus diseases recorded in the previous decade. Indeed, at the beginning of fruiting, the incidence of aphid borne viruses did not exceeded 1.6% of vines for the 29 shade houses which adopted the virus prophylaxis. The three remaining shade houses were severely (20–50%) infected by WMV or CymMV because of the planting of virus-infected cuttings. These results demonstrate the benefit of implementing prophylaxis at the scale of an archipelago, and widen the possibilities of developing intensive cultivation of vanilla in other areas confronted with similar virus constraints.     

云南鬼针草上发现鬼针草斑驳病毒

 【目的】鉴定侵染鬼针草引起花叶或斑驳的病原。【方法】利用电镜技术观测病原粒子;间接ELISA方法鉴定病原与马铃薯Y病毒属病毒的血清学关系;分子生物学技术克隆了病原3′-端序列;BLAST,Clustal_X,BioEdit和MEGA 4.0等生物信息学软件用于所得序列的序列分析。【结果】在电镜下可观察到长约650-700 nm×13 nm左右的线状病毒粒子和风轮状内含体;利用Potyvirus PathoScren试剂盒检测呈阳性;用马铃薯Y病毒科病毒简并引物可扩增获得一条约1 800 bp的片段;序列分析表明该序列与鬼针草斑驳病毒相应序列高度相似,外壳蛋白的氨基酸同源性及3′-UTR核苷酸同源性均达96%以上。【结论】侵染云南鬼针草引起花叶或斑驳症状的病原为鬼针草斑驳病毒,这是该病毒侵染中国鬼针草属植物的首次报道。     

Identification and characterization of a potyvirus causing chlorotic spots on <Emphasis Type="Italic">Phalaenopsis</Emphasis> orchids

A putative virus-induced disease showing chlorotic spots on leaves of Phalaenopsis orchids was observed in central Taiwan. A virus culture, phalaenopsis isolate 7-2, was isolated from a diseased Phalaenopsis orchid and established in Chenopodium quinoa and Nicotiana benthamiana. The virus reacted with the monoclonal antibody (POTY) against the potyvirus group. Potyvirus-like long flexuous filament particles around 12–15 × 750–800 nm were observed in the crude sap and purified virus preparations, and pinwheel inclusion bodies were observed in the infected cells. The conserved region of the viral RNA was amplified using the degenerate primers for the potyviruses and sequence analysis of the virus isolate 7-2 showed 56.6–63.1% nucleotide and 44.8–65.1% amino acid identities with those of Bean yellow mosaic virus (BYMV), Beet mosaic virus (BtMV), Turnip mosaic virus (TuMV) and Bean common mosaic virus (BCMV). The coat protein (CP) gene of isolate 7-2 was amplified, sequenced and found to have 280 amino acids. A homology search in GenBank indicated that the virus is a potyvirus but no highly homologous sequence was found. The virus was designated as Phalaenopsis chlorotic spot virus (PhCSV) in early 2006. Subsequently, a potyvirus, named Basella rugose mosaic virus isolated from malabar spinach was reported in December 2006. It was found to share 96.8% amino acid identity with the CP of PhCSV. Back-inoculation with the isolated virus was conducted to confirm that PhCSV is the causal agent of chlorotic spot disease of Phalaenopsis orchids in Taiwan. This is the first report of a potyvirus causing a disease on Phalaenopsis orchids.     

Occurrence of Telosma mosaic virus causing passion fruit severe mosaic disease in Thailand and immunostrip test for rapid virus detection

The cause of leaf and fruit severe mosaic disease in purple passion fruit (Passiflora edulis) in Chiang Mai province, Thailand, was identified and virus diagnostic kit was developed. Symptomatic plants collected from diseased mother plants in the nursery at the Royal Pangda research station (RPRS) was used for virus isolation through single lesion transfer in Chenopodium amaranticolor, followed by virus propagation for purification in Nicotiana benthamiana, and back inoculation to purple passion fruit. Electron microscopy of leaves with typical symptoms revealed potyvirus-like flexuous rod particles, ca. 750 nm long and pinwheel inclusion bodies in the cytoplasm of infected cells. The deduced amino acid sequence of the complete coat protein revealed that the isolated virus was a strain of Telosma mosaic virus with which it shared 84% identity. A rabbit polyclonal antiserum was produced against purified virions, and used to develop a gold-labeled immunostrip for rapid virus diagnosis. The test strip could detect the virus in diseased passion fruit sap up to a dilution of 1: 640, and positive test line could be read within 3–5 min. Application of strip test for virus assay in RPRS nursery's mother plants help screen clean stocks. This strip test kit supports RPRS program of producing virus-free planting materials for farmers.     

辣椒脉斑驳病毒文昌分离物基因组测序及分析

辣椒脉斑驳病毒（Chilli veinal mottle virus,ChiVMV）近年来严重危害海南黄灯笼辣椒的产量,开展针对该病毒的研究以求达到对其有效防控迫在眉睫。本研究分7段克隆了ChiVMV文昌分离物（ChiVMV Wenchang isolate,ChiVMV-WC）的基因组。分段克隆的测序结果通过拼接,共获得ChiVMV-WC基因组9717个核苷酸（nt）序列（GenBank登录号：GQ981316）。在核苷酸序列的第164位～第9270位存在一个大的开放阅读框（ORF）,编码一个多聚蛋白（分子量为350.44kD）。近年来被证实的马铃薯Y病毒科的一个新的ORF（pretty interesting potyviridae ORF,PIPO）存在于ChiVMV-WC基因组核苷酸序列的第2892位～第3110位,编码一个由72个氨基酸聚合而成的多肽（分子量为8.26kD）。ChiVMV-WC与ChiVMV其它分离物的基因组核苷酸序列比较发现该病毒存在着较高的变异率。本研究通过与同属内病毒的同源性分析以及系统进化树分析,确定了ChiVMV-WC在生物进化史上的地位。