The S RNA segment determines symptom differences on Tetragonia expansa between two Watermelon silver mottle virus isolates

The WS-Y isolate of Watermelon silver mottle virus (WSMoV) causes severe necrosis in Tetragonia expansa. To determine the RNA segment that induces symptoms, genome reassorants between WS-Y and an isolate causing mild mottle, WS-O, were generated. The origin of each segment in the reassortants was identified by RT-PCR and subsequent restriction enzyme analysis of the amplified fragments. Thirty genome reassortants were isolated from co-infected T. expansa plants. The reassortants with the S RNA segment of WS-Y caused severe necrosis, while those with the S RNA segment of WS-O caused a mild mottle; hence, the S RNA determined symptom expression. The incidence of reassortants was disproportional among genotypes. The most frequent genome reassortant possesses the L RNA of WS-Y, the M RNA of WS-O and the S RNA of WS-Y. A similar ratio of genotypes was found in isolates of local lesions on Chenopodium quinoa. These results strongly suggested that competition occurred independently between the individual RNA segments in a co-infected T. expansa plant, not between isolates.     

新疆番茄病毒病检测及南方番茄病毒全基因组序列测定与传播特性分析

为了解新疆番茄上病毒病的发生情况,利用一步法RT-PCR技术检测了南北疆番茄上南方番茄病毒(Southern tomato virus,STV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、番茄花叶病毒(Tomato mosaic virus,ToMV)以及马铃薯Y病毒(Potato virus Y,PVY)的感染情况,并利用分段克隆的方法进行全基因组测序,通过RT-PCR方法检测健康植株与携带病毒植株杂交育种的F1代植株带毒率以分析STV的种子传播特性。结果显示,新疆番茄上CMV、STV、ToMV和PVY在北疆的检出率分别为52%、37%、27%和14%;在南疆的检出率分别为79%、60%、69%和0;且以STV、CMV及ToMV的复合侵染为主。从我国加工番茄上首次获得了长3 437 nt的STV SHZ-1核苷酸序列,序列比对分析发现其与已报道STV只有1~9个核苷酸的变异,且序列变异与地域无相关性。分析杂交F1代加工番茄植株上STV的传播特性,发现其除可由种子传播外,也可通过花粉传播。表明STV是侵染新疆番茄的主要病毒之一,且该病毒可通过种子或杂交育种途径进行传播。     

Effects of foliar sprayed calcium sources onTomato mosaic virus (ToMV) infection in tomato plants grown in greenhouses

Spray solutions containing 0.3% Ca which were prepared from four different calcium sources were foliar-sprayed on greenhouse-grown tomato plants, infected with theTomato mosaic virus Tobamovirus (ToMV) or not. ToMV-infected and uninfected control groups were sprayed with distilled water. Growth and macronutrient (N, P, K, Ca and Mg) composition of tomato plants as well as virus concentration and its relative infectivity were investigated in treated and untreated plants. The Ca sprays were applied three times: on the same day as inoculation, and 15 and 30 days after inoculation. Virus concentration in tomato plants generally decreased with foliar-sprayed Ca. Virus concentration (DAS-ELISA absorbance) was reduced by foliar-sprayed Ca, but plants remained infected. At the same time, tissue Ca concentrations increased significantly with foliar-applied Ca, with the exception of CaNO₃·4H₂O+0.05M Na-EDTA. ToMV reduced the fresh and dry weights and Ca concentrations of tomato plants, but significantly raised P concentration in the tissue. Neither virus inoculation nor foliar Ca applications affected N and Mg concentrations in tomato plants. The foliar-applied Ca from all the sources gave K concentrations similar to those of control plants. http://www.phytoparasitica.org posting Jan. 26, 2007.     

黄瓜绿斑驳花叶病毒基因组全序列分析及病害的田间调查

为明确近年来在浙江省葫芦科作物上发生的黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)基因组特征及其发生分布情况,从浙江省及上海地区的甜瓜、西瓜和瓠瓜上采集疑似样品进行RT-PCR鉴定,通过分段扩增测序的方法拼接获得基因组全序列并进行系统进化分析,利用特异性引物扩增获得CGMMV外壳蛋白(coat protein,CP)基因序列,制备CGMMV CP抗血清进行Western-blot和Dot-ELISA检测。结果显示,来自甜瓜、西瓜和瓠瓜的3个CGMMV分离物基因组全序列均具有烟草花叶病毒属典型基因组结构特征,全部由6 423 nt构成;3个全序列间的核苷酸同源性高达99.11%~99.67%,编码的CP氨基酸同源性为100%。系统进化分析发现,CGMMV不同分离物形成2个进化相关群体,3个浙江的CGMMV分离物均位于第I组内,与已报道的中国CGMMV分离物和韩国CGMMV分离物亲缘性较高。Western-blot检测表明CGMMV CP抗血清可以与感病植株中的病毒发生特异性反应,可用于CGMMV鉴定;Dot-ELISA检测发现CGMMV在浙江省和上海市的葫芦科作物上普遍存在。     

世界性害虫番茄潜麦蛾入侵我国的风险分析

为评估原产于南美洲的世界性毁灭害虫——番茄潜麦蛾Tuta absoluta(Meyrick)入侵我国的可能性和风险,应用有害生物风险分析方法对其进行风险评估,即对番茄潜麦蛾的不同入侵阶段风险进行定性分析,并应用多指标综合评价方法,从其国内外的发生现状、潜在的经济危害性、受害作物的经济重要性、入侵我国的可能性以及风险管理的难度等方面进行入侵风险的半定量分析。结果表明,番茄潜麦蛾虽然目前在我国还没有分布,但在我国大部分番茄产区都可以存活和定殖,是一种具有高度潜在威胁的有害生物;其入侵我国的风险评估值R为2.64,属于极高风险性农业有害生物。故此建议将其增补列入《中华人民共和国进境植物检疫性有害生物名录》,在新疆、云南、广西等边境省区开展预防性监测,并加强对该种害虫的检验检疫力度,严防其传入我国。     

Rapid Identification of Clavibacter michiganensis Subspecies Sepedonicus based on the Stable Low Molecular Weight RNA (LMW RNA) Profiles

Clavibacter michiganensis subsp. sepedonicus causes potato ring rot disease. The identification process for this bacterium is complex and long. This work demonstrates that the stable low-molecular-weight (LMW) RNA profiles allow their rapid identification. Staircase electrophoresis in polyacrylamide gels was used to analyze the LMW RNA profiles of 54 strains of C. michiganensis subsp. sepedonicus from different geographic origins. The profiles of several strains of other subspecies of C. michiganensis and other pathogens of potatoes were also analyzed. All the strains of C. michiganensis subsp. sepedonicus had the same LMW RNA profile. They had a band in class 2 of tRNA that was absent in the other subspecies of the species C. michiganensis. Also, the LMW RNA of C. michiganensis subsp. sepedonicus was different with respect to the LMW RNA profiles of other pathogens of potato. The results indicate the possible utilization of LMW RNA profiles in identification of the bacteria causing potato ring rot disease.     

Common reed (Phragmites communis) is a natural host of important cereal viruses in the Trakya region of Turkey

Common reed (Phragmites communis Trin.), a perennial grass, is a widespread weed in the Trakya region of Turkey. Reed leaf samples were collected in 2004 and 2005, and tested for the presence of theMaize dwarf mosaic virus (MDMV),Sugarcane mosaic virus (SCMV),Barley yellow dwarf virus-PAV (BYDV-PAV),Cereal yellow dwarf virus-RPV (CYDV-RPV) andWheat dwarf virus (WDV) by DAS-ELISA, PTA-ELISA and Western blot analysis. MDMV was identified in five out of sixP. communis samples that exhibited characteristic virus-like symptoms in 2004. The remaining sample was co-infected with MDMV and BYDV-PAV. Transmission electron microscopy confirmed the presence of flexuous rod-shaped virus particles in four samples that reacted positively for MDMV in ELISA. In 2005, ELISA revealed that nine out of 234 samples that were collected in two different locations were infected with MDMV, nine with SCMV, and three with BYDV-PAV. No sample contained CYDV-RPV, JGMV and WDV. Our results confirm that the common reed is a host of BYDV-PAV and indicate, for the first time, that it is also a natural host of MDMV and SCMV.P. communis most likely acts as a reservoir of these three viruses in the Trakya region in Turkey. http//www.phytoparasitica.org posting Sept. 13, 2006.     

Characterization of a pothos (Scindapsus aureus) virus with unusual properties

A virus for which the name of pothos latent virus (PoLV) is proposed, was isolated by inoculation of sap from symptomless plants ofScindapsus aureus. PoLV had isometric particlesc. 30 nm in diameter, a monopartite genome consisting of a non polyadenylated, single-stranded RNA moleculec. 4,300 nucleotides in length, constitutingc. 17% of the particle weight, and a single type of coat protein subunit with aM _r ofc. 40,000 Daltons. The biological properties (host range reactions) of PoLV resembled those ofTombusviridae for it infected most of the artificial hosts locally, inducing symptoms recalling those elicited by several species of the above family. Like tombus- and carmoviruses, PoLV had two subgenomic RNAs which, however, differed in size from those of both genera. The dsRNA pattern was also distinctly different. Cytopathological features recalled those of tombusviruses except for the lack of multivesicular inclusion bodies. PoLV was serologically related to, but distinct from twoCarmovirus (i.e., galinsoga mosaic and Ahlum waterborne viruses) and threeTombusvirus species (i.e. eggplant mottled crinkle, Sikte waterborne and Lato river viruses). Thus, PoLV had properties somewhat intermediate between those ofTombusvirus andCarmovirus genera but bridged the two taxa through the serological relationship with some of their species. The taxonomic position of PoLV is still undetermined. It must await the results of molecular investigations now underway.     

白菜型冬油菜NCED3基因、启动子的克隆及其表达分析

从2个不同抗寒性的冬油菜品种中克隆了NCED3基因及其启动子序列,并分析其在叶片和根中的表达,研究NCED3基因在冬油菜中的作用机理。结果表明,陇油6号的NCED3基因开放阅读框(ORF)长度为1 794 bp,编码597个氨基酸,分子量65.74 kD,理论等电点为5.81;天油2号的ORF与其长度相同,分子量为65.78 kD,理论等电点为5.94。2个蛋白都是亲水蛋白,具有疏水峰。根据预测,启动子具有生物过程中常见的顺式作用元件如CAAT-box等、分生组织表达CAT-box相关的顺式作用元件、-30 TATA box附近的核心启动子元件等,还有ABRE、TGA-element、CGTCA-motif、TGACG-element等激素响应元件,此外,还鉴定出低温响应元件（LTR）。2个品种启动子序列的相似度为99.38%,只有1个不同的顺式作用元件,即circadian,推测其与昼夜节律相关。低温、PEG及ABA处理后NCED3在叶片和根中的表达均高于对照,增加范围为0.07~6.48,且均于8 h达到峰值。与根系的表达特性相比,胁迫处理后叶片的基因表达均在2 h显著升高,天油2号的升高幅度（1.54~6.00）均大于陇油6号（0.04~1.95）。     

Association of the western flower thrips (Frankliniella occidentalis) with cultivated sunflower (Helianthus annuus) in Israel

In 1992 and 1993, sunflower (Helianthus annuus L.) crops in Israel were heavily damaged by the western flower thrips (WFT),Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). Adult females appear in the heads of sunflower just as the latter begin to blossom; the population decreases towards completion of anthesis. The damage to kernels is greatest at the periphery of the heads, where flowers appear first, and lowest at the center. Fields sown early in the season (in March) are usually the ones infested most by WFT and suffering the greatest damage. Fields sown one month later are less infested, since predatory bugs of the genusOrius are very active at that time.     

梨小食心虫谷胱甘肽S-转移酶基因序列分析及其与吡虫啉的相关性

为明确梨小食心虫Grapholita molesta谷胱甘肽S-转移酶（glutathione S-transferase,GST）对吡虫啉的代谢作用,基于不同亚致死浓度吡虫啉处理的梨小食心虫转录组数据库进行筛选,并通过生物信息学分析、系统发育树构建、保守位点序列比对、分子对接和表达谱分析来确定梨小食心虫代谢吡虫啉过程中的关键GST。结果显示,共筛选到梨小食心虫的17个GST基因,其中GmGSTS1、GmGSTD2、GmGSTE5和GmGSTE4基因编码的蛋白与苹果蠹蛾Cydia pomonella的GST蛋白有较高的相似性,相似度达69.12%～89.66%。梨小食心虫GmGSTD2蛋白G位点和H位点的保守氨基酸与褐飞虱Nilaparvata lugens及烟粉虱Bemisia tabaci代谢吡虫啉相关GST氨基酸序列完全一致。吡虫啉与梨小食心虫GmGSTD2蛋白G位点的ARG-66和H位点的TYR-112可以形成稳定的氢键;与GmGSTE3蛋白G位点的LEU-45和H位点的LYS-123可以形成稳定的氢键。在吡虫啉处理后,梨小食心虫体内GmGSTD2和GmGSTE3基因的表达量显...     

Characterization of isolates of binucleate Rhizoctonia spp. associated with strawberry black root rot complex using fatty acid methyl ester (FAME) profiles

Black root rot is an important disease of strawberry caused by a complex of fungi that includes species of Rhizoctonia. In this study, a modified MIDI method (Microbial Identification System) was investigated for its utility to differentiate isolates of the three different anastomosis groups (AGs) of binucleate Rhizoctonia spp., associated with strawberry black root rot complex representing AG-A, AG-G, and AG-I. A total of 11 fatty acids were detected, and the FAME profiles for isolates of the three different AGs of Rhizoctonia spp. varied quantitatively and qualitatively. Moreover, the modified MIDI method will be a useful discriminatory tool for fungal identification and classification of the AGs of binucleate Rhizoctonia spp. associated with strawberry black root rot complex.     

Shoot meristem tissue of tobacco inoculated with Cucumber mosaic virus is infected with the virus and subsequently recovers from infection by RNA silencing

Distribution of Cucumber mosaic virus (CMV) in shoot meristem tissue of CMV-inoculated tobacco was successively analyzed with immunohistochemical microscopy and in situ hybridization. CMV signals were detected in the tissue at 7 days postinoculation (dpi), but then they decreased and disappeared after 14dpi. Detailed observation confirmed CMV invasion of shoot apical meristem at 6–8dpi. Short interfering RNA corresponding to CMV RNAs was first detected at 7dpi and was detected up to 24dpi. These results suggest that the shoot meristem tissue is infected with CMV but subsequently recovers from the infection by RNA silencing.     

甜菜夜蛾中肠碱性磷酸酶alp2基因的克隆、表达及功能分析

为探究甜菜夜蛾Spodoptera exigua中肠碱性磷酸酶(alkaline phosphatase protein 2,ALP2)是否为Cry1Ac杀虫蛋白的受体,采用同源克隆和RACE技术克隆了编码alp2基因的完整c DNA序列,利用荧光定量PCR比较了甜菜夜蛾幼虫中肠不同龄期ALP2表达量的差异,利用Ligand blot方法检测了中肠ALP2与Cry1Ac杀虫蛋白的结合。结果表明,alp2基因序列全长1 629 bp(Gen Bank序列号为KP420013),编码542个氨基酸,预测在氨基酸序列N端包含1个由21个氨基酸组成的信号肽,在C端存在1个GPI修饰的锚定位点,且在整个氨基酸序列中存在多个糖基化修饰位点。在整个甜菜夜蛾幼虫期均有ALP2表达,但不同龄期的表达量差异显著,1龄幼虫期表达量最低,4龄幼虫期最高。Ligand blot方法检测结果表明原核表达的ALP2片段与活化的Cry1Ac杀虫蛋白可以结合。研究表明,甜菜夜蛾中肠的ALP2可能是Cry1Ac的受体之一。     

Further identification of races of Cladosporium fulvum (Fulvia fulva) on tomato originating from the Netherlands France and Poland

Races ofCladosporium fulvum, which can overcome the resistance of the genes Cf₂, Cf₄, Cf₅, Cf₈, Cf₉ and Cf₁₁ have appeared in the Netherlands, France and Poland. Known isolates from the Netherlands and France and three new isolates from Poland have been investigated for the presence of virulence genes using a set of genotypes carrying resistance genes Cf₂ to Cf₁₁.Several Dutch isolates of races, earlier designated as 2.4, 2.4.5 and 2.4.5.9, were found to break down the resistance gene Cf₁₁. These races must therefore be designated as 2.4.11, 2.4.5.11 and 2.4.5.9.11 respectively. In the new Polish isolates virulence genes, overcoming the resistance genes Cf₂, Cf₄, Cf₈, Cf₉ and Cf₁₁ were found. Since all races able to grow on genotypes with Cf₄, could also grow on genotypes carrying Cf₈, it was impossible to discriminate between the genes Cf₄ and Cf₈. These Polish isolates were designated as races 4.11, 2.4.11 and 2.4.9.11. The consequences of the occurrence of these races for tomato breeding are discussed.Samenvatting Fysio's vanCladosporium fulvum, die de resistentie-genen Cf₂, Cf₄, Cf₅, Cf₈, Cf₉ en Cf₁₁ kunnen doorbreken, zijn in Nederland, Frankrijk en Polen opgetreden. Met behulp van een groep genotypen, die de resistentie genen Cf₂ tot en met Cf₁₁ dragen, zijn Nederlandse, Franse en enkele nieuwe Poolse isolaten onderzocht op de aanwezigheid van virulentiegenen. Enkele Nederlandse isolaten, eerder aangeduid met 2.4, 2.4.5. en 2.4.5.9, bleken het resistentie-gen Cf₁₁ te kunnen doorbreken. Deze moeten daarom aangeduid worden als respectievelijk 2.4.11, 2.4.5.11 en 2.4.5.9.11. In de nieuwe Poolse isolaten werd virulentie gevonden voor Cf₂, Cf₄, Cf₈, Cf₉ en Cf₁₁. Alle fysio's die op genotypen met Cf₄ konden groeien, groeiden ook op genotypen met Cf₈. Daarom kon geen ondersheid gemaakt worden tussen Cf₄ en Cf₈. De Poolse isolaten behoren tot de fysio's 4.11, 2.4.11 en 2.4.9.11. De gevolgen van het voorkomen van deze fysio's voor de tomateveredeling worden besproken.