Complete Genome Sequence of a Chinese Isolate of Melon Necrotic Spot Virus and its Phylogenetic Relationship

The full‐length nucleotide sequence and genomic organization of a melon necrotic spot virus isolate from Haimen, China (MNSV‐HM), were determined. The MNSV‐HM genome consists of a positive sense single‐stranded RNA, 4267 nt in length, with at least five open reading frames (ORFs) encoding p29, p89, p42, and two small 7‐kDa proteins (p7A and p7B). p89 shares a common start codon with p29 and continues through the amber stop codon of p29 to produce an 89‐kDa protein. The p7A ORF terminates in an amber codon whose read‐through could generate a 14‐kDa protein. Phylogenetic analyses based on the p42 amino acid sequence and complete genomic sequence placed MNSV‐HM and Spanish isolates of the virus in a major cluster, indicating a close genetic relationship. In conclusion, we report the full‐length sequence of MNSV‐HM and its translation strategy. The obtained genomic organization and phylogenetic trees indicate that MNSV‐HM belongs to the MNSV genus Carmovirus. To the best of our knowledge, this is the first demonstration of the complete nucleotide sequence of an MNSV isolate from China.     

Occurrence of Melon Necrotic Spot Virus in Crete (Greece)

Since 1982 melon plants cv. Galia grown in plastic houses showed severe leaf and stem necrosis. A virus isolated from affected plants infected only Cucurbitaceae. Systemic infections were only developed in melon and Luffa acutangula, while a severe hypocotyl necrosis was observed in watermelon. Purified virus sedimented in sucrose gradients as a single component and reacted only with antisera to melon necrotic spot virus (MNSV). Seed transmission (22.5%) of the virus was observed m melon plants grown from seed of naturally infected melons, but the virus could not be detected in triturated seeds. The virus could be isolated from leachate of contaminated soil and melon plants became infected when grown in contaminated soil or when watered with suspensions of virus. These properties suggest that the virus is an isolate of MNSV.     

福州地区对虾白斑病病毒的超微结构

在福州地区分离到一种高致病性的白斑病病毒 ,该病毒仅存在于细胞质中 ,完整的病毒粒子有囊膜 ,一端略圆 ,一端稍尖 ,直径约为 80～ 10 0nm ,囊膜与核衣壳之间的间隙约为 2 0～ 2 5nm。该病毒在细胞内不形成包含体 ,但有些可形成封入体。负染观察到的病毒核衣壳呈直杆状 ,但长度和直径相差较大 ,最长的病毒超过 6 0 0nm。这些特征与其它已报道的白斑病病毒有所不同 ,因此暂将它称为“对虾白斑病病毒福州分离株”。     

福州地区对虾白斑病病毒的超微结构

在福州地区分离到一种高致病性的白斑病病毒,该病毒仅存在于细胞质中,完整的病毒粒子有囊膜,一端略圆,一端稍尖,直径约为80～100 nm,囊膜与核衣壳之间的间隙约为20～25 nm.该病毒在细胞内不形成包含体,但有些可形成封入体.负染观察到的病毒核衣壳呈直杆状,但长度和直径相差较大,最长的病毒超过600 nm.这些特征与其它已报道的白斑病病毒有所不同,因此暂将它称为"对虾白斑病病毒福州分离株".     

Characterization of an Isolate of Tobacco Necrosis Virus From Zucchini

Tobacco necrosis virus (TNV) was isolated from glasshouse zucchini plants in Liguria, northern Italy, showing yellow spots on young leaves and necrotic symptoms on older leaves, petioles and stems. No other viruses were present in these plants. Experimentally, the virus systemically infected zucchini seedlings grown in naturally infected soil but only local infections were induced when leaves and roots were mechanically inoculated. The virus was identified as serotype D, devoid of satellite particles. This appears to be the first report of TNV in zucchini. Three ELISA procedures for field detection of TNV using polyclonal antibodies for serotype D were compared. Antibodies were conjugated with I) alkaline phosphatase (AP-AB);2) N-Hydroxy-succinimid obiotin (BNHS-Ab); 3) N-biotinyl-6-aminocaproic hydrazide (BACH-Ab). The procedure using BNHS-Ab detected the lowest concentration of the homologous virus, followed by BACH-Ab and AP-Ab. Only with biotinylated antibodies. TNV serotype A was also detected.     

Natural Occurrence of Helenium Virus S in Impatiens holstii

Samples of commercially grown Impatiens holstii from several sources in Minnesota contained a carlavirus which serologically was closely related, if not identical with Helenium virus S described earlier in Germany. The two viruses have similar experimental host ranges and caused similar cytopathogenic effects. After mechanical transmission the Impatiens isolate produced less severe symptoms in Impatiens than the Helenium isolate. Only the latter isolate was transmitted by Myzus persicae in the non-persistent manner.     

韭菜病毒分离物初步鉴定

韭菜病毒分离物初步鉴定房德纯,王振东,佟成富,陆庆轩（沈阳农业大学植保系,沈阳１１０１６１）（辽宁省风沙地改良利用研究所,阜新１２３０００）（沈阳市园林科学研究院,沈阳１１００１５）关键词韭菜病毒病,毒原鉴定,韭菜萎缩病毒１９９３年在辽宁省阜新市韭菜...     

A Hamster-Derived West Nile Virus Isolate Induces Persistent Renal Infection in Mice

Background

West Nile virus (WNV) can persist long term in the brain and kidney tissues of humans, non-human primates, and hamsters. In this study, mice were infected with WNV strain H8912, previously cultured from the urine of a persistently infected hamster, to determine its pathogenesis in a murine host.

Methodology/Principal Findings

We found that WNV H8912 was highly attenuated for neuroinvasiveness in mice. Following a systemic infection, viral RNA could be detected quickly in blood and spleen and much later in kidneys. WNV H8912 induced constitutive IL-10 production, upregulation of IFN-β and IL-1β expression, and a specific IgM response on day 10 post-infection. WNV H8912 persisted preferentially in kidneys with mild renal inflammation, and less frequently in spleen for up to 2.5 months post infection. This was concurrent with detectable serum WNV-specific IgM and IgG production. There were also significantly fewer WNV- specific T cells and lower inflammatory responses in kidneys than in spleen. Previous studies have shown that systemic wild-type WNV NY99 infection induced virus persistence preferentially in spleen than in mouse kidneys. Here, we noted that splenocytes of WNV H8912-infected mice produced significantly less IL-10 than those of WNV NY99-infected mice. Finally, WNV H8912 was also attenuated in neurovirulence. Following intracranial inoculation, WNV persisted in the brain at a low frequency, concurrent with neither inflammatory responses nor neuronal damage in the brain.

Conclusions

WNV H8912 is highly attenuated in both neuroinvasiveness and neurovirulence in mice. It induces a low and delayed anti-viral response in mice and preferentially persists in the kidneys.     

Properties of the Iranian Isolate of Bermudagrass Etched-line Virus

The Iranian isolate of bermudagrass etched-line virus (BELV-I), purified by low-pH treatment of infected bermudagrass sap followed by several cycles of differential centrifugation and sucrose density-gradient centrifugation, formed two components in density-gradient columns. The top component consisted of empty protein shells. It had a major structural protein of c. 22 kDa and a minor of c. 25 kDa. The weight of the nucleic acid present only in bottom component particles, was calculated to be 1.82 × 10⁶ Da. Only Aconurella prolixa (Leth.) was able to transmit the virus under experimental conditions or contained the virus in natural populations. In ELISA tests the virus titer in the vector increased rapidly between days 14 and 29 after acquisition, results indicating a propagative relationship. BELV-1 was serologically closely related to the Moroccan isolate of BELV, and related to the American but not to the Costa Rican isolate of maize rayado fino virus (MRFV). Several graminaceous species were found to be experimental or natural hosts of the virus.     

西瓜花叶病毒中国分离株全基因组核苷酸序列测定

西瓜花叶病毒(Watermelon mosaic virus,WMV)是马铃薯Y病毒属(Potyvirus)成员,主要危害西瓜和甜瓜,引起花叶病。在田间,该病害主要由蚜虫以非持久性方式传播。西瓜和甜瓜花叶病在国内陕西、山东、云南、辽宁、山西、新疆、河南和黑龙江等地广泛发生[1-6]。从20世纪80年代中期开始发生,逐渐上升为普遍发生的主要病害。我国大部分地区因西瓜和甜瓜病毒病造成的损失为30%~50%,甚至会绝产,西瓜花叶病毒已经成为制约西瓜和甜瓜高产稳产最主要的因素之一[7]。到目前为止,多数工作集中在对西瓜和甜瓜病毒病的鉴定,在分子生物学上仅限于对CP基因…     

热带不同生境稀有放线菌分离

通过物理和化学方法对海南清澜港红树林、海南东寨港红树林、湛江红树林、深圳红树林、吊罗山原始森林、儋州橡胶林和海口火山口等7处热带不同生境土壤样品预处理,共分离到114株放线菌。用显微镜形态观察和菌落形态观察对分离到的放线菌初步归类,从中选取13株菌进行16SrDNA序列分析,并构建系统发育树进行类群。由初步的归类结果对热带不同生境稀有放线菌的类群分布进行分析比较,水生环境和非水生环境以及不同的非水生环境之间,发现稀有放线菌类群分布有着明显差异。     

A New Virulent Isolate of Clover Yellow Vein Virus on Phaseolus vulgaris in Bulgaria

Potyviruses are a common threat for snap bean production in Bulgaria. During virus surveys of bean plots in the south central region, we identified an isolate of Clover yellow vein virus (ClYVV), designated ClYVV 11B, by indirect ELISA and RT‐PCR causing severe mosaic symptoms and systemic necrosis. Indirect and direct ELISA using ClYVV antisera differentiated the ClYVV isolate from Bean yellow mosaic virus (BYMV), but serological analysis could not distinguish the Bulgarian isolate ClYVV 11B from an Italian ClYVV isolate used as a reference (ClYVV 505/7). RT‐PCR analyses with specific primers revealed that both isolates were ClYVV. Sequence analysis of an 800 bp fragment corresponding to the coat protein coding region showed 94% identity at the nucleotide level between the two isolates. Phylogenetic analyses of aligned nucleotide sequences available in the database confirmed the existence of two groups of isolates, but ClYVV 11B and ClYVV505/7 belonged to the same group. We compared the virulence of both isolates on a set of differential cultivars and 19 bean breeding lines resistant to Bean common mosaic virus (BCMV) and Bean common mosaic necrosis virus (BCMNV): Bulgarian isolate ClYVV 11B was able to infect systemically all tested bean differential cultivars and breeding lines including those with genotypes Ibc3 and Ibc2²; Italian isolate ClYVV 505/7 was not able to infect systemically some differentials with genotypes bc‐ubc1, bc‐ubc2², bc‐ubc2bc3, Ibc1², Ibc2², Ibc3. The role of bc3 gene as a source of resistance to potyviruses is discussed.     

Purification and Characterization of a Jordanian Isolate of Watermelon Mosaic Virus

The Jordanian isolate of watermelon mosaic virus-2 (WMV-2Jo) was purified from infected Cucurbita pepo cv. Top Capi by extraction in 0.05 M sodium citrate buffer containing 0.01 M sodium diethyl dithiocarbamate and 0.01 M cysteine hydrochloride (0.01 M D + C), clarification with chloroform and n-butanol, sedimentation by ultracentrifugation, and rate-zonal centrifugation in 10–40% sucrose gradient. The purified virus had an ultraviolet absorption spectrum typical of a nucleoprotein with a low nucleic acid content. Homologous antiserum had a titre of 1: 256, as determined by the ring interface test. Electron microscopy of negatively stained purified virus revealed flexuous particles with a normal length of 750 nm. Cytoplasmic spindle-shaped inclusions were readily detectable in infected epidermal cells under the light microscope. Thin sections of infected tissue revealed the presence of laminated aggregates, pinwheel and scroll inclusions. The virus reacted with antisera prepared to the Florida strain of WMV-2 and zucchini yellow mosaic virus in a sodium dodecyl sulfate (SDS) agar gel diffusion test. Using the Derrick-decoration combined technique of immune electron microscopy, the virus reacted strongly with the homologous antiserum and zucchini yellow mosaic virus antiserum, but less with antiserum prepared to the Florida strain of WMV-2.     

莴苣花叶病毒浙江余杭分离物基因组全序列及其结构分析

测定了莴苣花叶病毒(LMV)浙江余杭分离物基因组全序列.此病毒分离物基因组由10*!080个核苷酸组成,具典型的马铃薯Y病毒科成员基因组结构,与已报道的欧洲、美国和巴西LMV核苷酸同源性为96.7%～98.8%,氨基酸同源性97.8%～99.0%.根据外壳蛋白氨基酸序列和5′端非编码区核苷酸序列分析比较及分子进化树分析,可将全球LMV分为西欧-加利福尼亚、希腊和也门3个类群.LMV有可能起源于加利福尼亚州向西欧,进而向希腊和也门扩展,浙江余杭LMV有可能来源于加利福尼亚州和西欧.