北京引致大丽菊花叶症的三种病毒

 大丽菊(Dahlia pinnata Cav.)花叶病已成为花卉生产出口的严重障碍。作者于1986-1987年在表现花叶的北京大丽菊上分离到两种形态大小不同的病毒分离物,电镜检测到一直径较大的球形病毒,分别定为BDM-1、BDM-2和BDM-3。经寄主反应、体外抗性、粒体形态、血清学等鉴定,病毒分离物BDM-1是黄瓜花叶病毒CMV;BDM-2是烟草花叶病毒TMV;BDM-3是大丽菊花叶病毒DaMV。从寄主反应、粒体大小、衣壳蛋白氨基酸组份及沉降特性综合分析看,BDM-1可能是CMV的另一株系CMV-DP。     

苜蓿盲蝽在豫东棉区的寄主选择及其转移规律

采用扫网、目测、盘拍和室内饲养方法,调查研究了河南东部棉区苜蓿盲蝽的野生寄主和栽培寄主植物种类,已发现161种,其中野生寄主植物137种,分属32个科,栽培寄主20种。研究了苜蓿盲蝽在这些寄主植物间的转移规律,表明该棉区以田菁、草木樨、野苜蓿为主的豆科植物及小白酒草为主的菊科等植物和小麦、棉花一起,构成了有利于苜蓿盲蝽发生的生态环境。抽穗期的小麦为越冬代成虫向棉田过渡的主要桥梁寄主,棉花仅仅是第1至第3代苜蓿盲蝽的侨居寄主,田菁、草木樨、小白酒草等豆科和菊科杂草丛生的地方是其第4代、第5代生存和越冬的主要场所。     

红颈常室茧蜂对绿盲蝽及其为害寄主植物的选择行为

为明确红颈常室茧蜂Peristenus spretus Chen et van Achterberg定位寄主绿盲蝽Apolygus lucorum(Meyer-Dür)时利用的挥发性信息物质来源,利用Y型昆虫嗅觉仪测定了红颈常室茧蜂雌雄成虫对绿盲蝽3龄若虫及其为害寄主植物棉株以及雌性红颈常室茧蜂成虫、绿盲蝽成虫对苗期和花期的棉花、蓖麻的选择行为。结果表明,与干净空气(空白对照)相比,雌性红颈常室茧蜂对绿盲蝽3龄若虫、健康棉株的选择行为无显著差异,绿盲蝽3龄若虫为害的棉株对雌性红颈常室茧蜂具有显著的吸引作用,但在为害后去虫和未去虫处理间无显著差异;雄性红颈常室茧蜂对各处理均无明显的选择行为。与健康或为害后去虫的苗期棉花相比,雌性绿盲蝽成虫显著趋向花期蓖麻;而雌性红颈常室茧蜂只趋向于为害后的花期蓖麻。雌性红颈常室茧蜂对为害后去虫的花期棉花和花期蓖麻的选择行为显著高于相应的苗期植株。这表明雌性红颈常室茧蜂主要利用绿盲蝽若虫为害后寄主植物释放的挥发性信息物质来定位寄主绿盲蝽,且其对花期寄主植物的选择行为与绿盲蝽成虫具有一致性。     

我国部分地区常见农作物上黄瓜花叶病毒分离物核酸多样性分析

为明确我国黄瓜花叶病毒株系分化及系统进化基本情况,从湖南、新疆、青海和海南4省区采集1 367个样品对其进行酶联免疫和RT-PCR检测,并对分离获得的15个黄瓜花叶病毒(Cucumber mosaic virus,CMV)纯化分离物CP、MP、2b核苷酸序列进行相似性和进化树分析及生物学性状比较。结果表明,辣椒、龙葵和黄瓜的CMV阳性检出率较高,分别为54.13%、29.19%和18.46%。进化树分析显示CMV-Q5与CMV亚组II的亲缘性较高;CMV-N7为新发现的重组株系,其CP、2b基因属于CMV亚组IB,MP基因却属于CMV亚组II;其余13个分离物均属于CMV亚组IB。CMV-N7和CMV-Q5在系统寄主心叶烟和枯斑寄主苋色藜上引发的症状相似,但比对照株系CMV-P3613(IB)的发病时间要晚1~2 d,系统花叶较温和,枯斑较小。表明在以上4省区常见农作物上广泛流行的CMV存在分子变异。     

烟草病毒病的防治方法

烟草病毒病害是烟草的主要病害.发病普遍严重,在我国部分烟区造成严重的损失,威胁着烟叶生产的可持续发展,已经成为烟叶优质、高效生产的限制因素.烟草病毒病的种类很多,已报道的有20多种,病原的个体很小,其基本形态为粒体,大部分植物病毒的粒体为球状、杆状和线状,少数为弹状、杆菌状和双联体状等.烟草植物病毒没有主动侵染寄主的能力,自然状态下主要靠介体和非介体传播.病毒的介体生物主要有蚜虫、叶蝉和飞虱,土壤中的线虫和真菌以及杂草等.病毒的非介体传播途径主要是通过机械、有性和无性繁殖材料、嫁接等方式.     

西瓜花叶病毒二号和黄瓜花叶病毒的新介体——伞形蚜的研究

伞形蚜Aphis umbrella(Borner)在我国尚未见报道。1980年在新疆五家渠一种锦葵科多年生植物上观察到一种蚜虫,传毒试验证明它可以传播西瓜花叶病毒二号(WMV—2)和黄瓜花叶病毒(CMV),从此,被列为瓜类病毒的主要介体之一。1985—1986年作者对该虫的生活周期、寄主、传毒习性及天敌进行了初步调查研究,简报如下。     

不同寄主植物对苜蓿盲蝽种群增长的影响

在自然变温条件下研究了4种寄主植物对苜蓿盲蝽生长、发育和繁殖的影响。结果表明:取食不同寄主植物的苜蓿盲蝽,其发育历期、存活率、若虫平均体重、成虫的繁殖力、寿命及各生命参数均有明显差异。供试植物中,苜蓿为嗜食寄主,棉花次之,莱豆和芝麻为非嗜食寄主。此研究为棉田合理的间套方式提供了依据。     

黄瓜花叶病毒外壳蛋白基因的克隆、原核表达及抗血清制备

黄瓜花叶病毒（Cucumber mosaic virus，CMV）是雀麦花叶病毒科Bromoviridae黄瓜花叶病毒属Cucumovirus的典型种。该病毒寄主范围广泛，能侵染85科365属1000多种植物。CMV除能单独侵染寄主植物外，还经常与TMV、PVY等复合侵染，给病害的田间调查和防治造成了困难。作者从山东青州的发病烟草植株上分离到了一个CMV青州分离物（命名为CMV-QZ），利用RT-PCR的方法克隆到了其外壳蛋白（coat protein，CP）基因，利用原核表达的CMV CP制备了抗血清，以期为CMV的准确快速检测提供依据。     

植物检疫性病毒传毒介体的传毒特征及检疫防控策略

本文在介绍植物病毒介体传毒一般性规律的基础上,总结了我国32种植物检疫性病毒有关的介体生物种类及其传毒特征,提出了介体内植物病毒的检测策略,可为口岸植物检疫性病毒的检测和检疫监管提供参考。     

四川省红叶石楠炭疽病病原菌鉴定及其潜在侵染源测定

为明确引起四川省红叶石楠炭疽病的病原菌及其潜在侵染源,采集疑似感染炭疽病的典型病叶进行分离获得纯化病原菌菌株,从中随机选取菌株HYSN3制成分生孢子悬浮液和菌饼,以无伤、刺伤、剪伤3种方式进行接种,筛选出效果最好的接种方式进行致病性测定,结合形态学特征与多基因序列分析将病原菌鉴定到种,并采用筛选出的接种方式将分离自其它19种寄主的23株炭疽菌接种到红叶石楠上,明确其潜在侵染源。结果表明,从红叶石楠病叶中共纯化得到14株菌株,基于形态特征和显微初步鉴定结果,从中选择8株代表菌株进行进一步鉴定。3种接种方式中,以刺伤后接种菌株HYSN3菌饼的效果最好,可用于致病性测定。基于形态学特征、致病性测定和多基因序列分析结果,将病原菌鉴定为胶孢炭疽菌Colletotrichum gloeosporioides(5株)、喀斯特炭疽菌C.karstii(1株)和暹罗炭疽菌C. siamense(2株),表明四川省红叶石楠炭疽病是由多种病原菌复合侵染引起的。来自其它寄主的23株炭疽菌菌株都能侵染红叶石楠,但致病力强弱不同,附近受炭疽菌侵染的植物都有可能成为红叶石楠炭疽病的潜在侵染源,园林植物养护过程中需予以一定的重视。     

Incidence of viruses in <Emphasis Type="Italic">Alstroemeria</Emphasis> plants cultivated in Japan and characterization of <Emphasis Type="Italic">Broad bean wilt virus-2, Cucumber mosaic virus</Emphasis> and <Emphasis Type="Italic">Youcai mosaic virus</Emphasis>

Alstroemeria plants were surveyed for viruses in Japan from 2002 to 2004. Seventy-two Alstroemeria plants were collected from Aichi, Nagano, and Hokkaido prefectures and 54.2% were infected with some species of virus. The predominant virus was Alstroemeria mosaic virus, followed by Tomato spotted wilt virus, Youcai mosaic virus (YoMV), Cucumber mosaic virus (CMV), Alstroemeria virus X and Broad bean wilt virus-2 (BBWV-2). On the basis of nucleotide sequence of the coat protein genes, all four CMV isolates belong to subgroup IA. CMV isolates induced mosaic and/or necrosis on Alstroemeria. YoMV and BBWV-2 were newly identified by traits such as host range, particle morphology, and nucleotide sequence as viruses infecting Alstroemeria. A BBWV-2 isolate also induced mosaic symptoms on Alstroemeria seedlings.     

Genetic analyses of Pseudomonas syringae isolates from Belgian fruit orchards reveal genetic variability and isolate-host relationships within the pathovar syringae, and help identify both races of the pathovar morsprunorum

A collection of Pseudomonas syringae and viridiflava isolates was established between 1993 and 2002 from diseased organs sampled from 36 pear, plum and cherry orchards in Belgium. Among the 356 isolates investigated in this study, phytotoxin, siderophore and classical microbiology tests, as well as the genetical methods REP-, ERIC- and BOX- (collectively, rep-) and IS50-PCR, enabled identification to be made of 280 isolates as P. syringae pv. syringae (Pss), 41 isolates as P. syringae pv. morsprunorum (Psm) race 1, 12 isolates as Psm race 2, three isolates as P. viridiflava and 20 isolates as unclassified P. syringae. The rep-PCR methods, particularly BOX-PCR, proved to be useful for identifying the Psm race 1 and Psm race 2 isolates. The latter race was frequent on sour cherry in Belgium. Combined genetic results confirmed homogeneities in the pvs avii, and morsprunorum race 1 and race 2 and high diversity in the pv. syringae. In the pv. syringae, homogeneous genetic groups consistently found on the same hosts (pear, cherry or plum) were observed. Pathogenicity on lilac was sometimes variable among Pss isolates from the same genetic group; also, some Psm race 2 and unclassified P. syringae isolates were pathogenic to lilac. In the BOX analyses, four patterns included 100% of the toxic lipodepsipeptide (TLP)-producing Pss isolates pathogenic to lilac. Many TLP-producing Pss isolates non-pathogenic to lilac and the TLP-non-producing Pss isolates were classified differently. Pseudomonas syringae isolates that differed from known fruit pathogens were observed in pear, sour cherry and plum orchards in Belgium.     

Genetic diversity and pathogenicity of Monilinia polystroma – the new pathogen of cherries

Brown rot caused by fungi belonging to the genus Monilinia is one of the major limiting factors of sour and sweet cherry production. Up to now, three species, M. fructigena, M. laxa and M. fructicola, have been identified as causal agents of brown rot on cherries worldwide. From 2010 to 2013, during the monitoring of cherry orchards in different areas of Poland, a fourth species, M. polystroma, was isolated from brown rot symptoms on sour and sweet cherry fruits. To the best of the authors’ knowledge, this is the first time M. polystroma has been reported as the causal agent of brown rot on cherries. The genetic diversity of M. polystroma isolates from cherries and other hosts was analysed using PCR MP, ISSR and RAPD techniques and showed its clear distinctness from other Monilinia spp. tested. The cluster analysis of fingerprinting data revealed a high similarity of M. polystroma isolates from Poland and their close relationship with the reference strain from Japan, indicating that this species is a recently introduced pathogen. The highest genetic distance between the examined isolates and the highest number of different genotypes was observed in an ISSR assay. Detailed genetic diversity characteristics revealed that M. polystroma isolates from cherries did not create a distinct group but were intermingled with M. polystroma isolates from other hosts. The results of the pathogenicity test conducted on different fruit species indicated a lack of host specificity for M. polystroma isolates.     

Biological and molecular characterization of two closely related carlaviruses affecting brassica plants

Carlaviruses are plant-infecting viruses with flexuous filamentous particles of approximately 650 nm in length and a positive single-stranded hexacistronic RNA molecule as the genome. In this study, we analysed 14 samples of brassicas plants, reportedly affected by carlaviruses, that were collected in distant and edaphoclimatic distinct regions in Brazil. The genomes of four viral isolates detected in leaf kale (Brassica oleracea var. acephala) plants displayed the typical genomic organization of carlaviruses, subdivided into two contrasting identity profiles. Isolates T25, T107, and T110 showed a higher sequence identity among them (c.93%) than with T90 (c.67%), which had the highest nucleotide sequence identity (85.2%) with the only documented genomic fragment, approximately 1 kb of the RNA 3′-end, of cole latent virus (CoLV). Identity values among the other three isolates and CoLV were consistently lower than 78.6%. Nucleotide and amino acid sequence identity values of the replicase cistron of the isolates T25 and T90 are below the threshold for species demarcation in the genus Carlavirus. Upon mechanical transmission, these two isolates induced different symptoms in brassicas and solanaceous plants. Overall, data indicated that these viruses belong to two related but distinct species of carlaviruses. While T90 was recognized as an isolate of CoLV, the remaining isolates were considered members of a new tentative carlavirus named Cole mild mosaic virus (CoMMV). Current work provides convincing support for the taxonomic status of the species Cole latent virus, enlarges the known diversity of carlaviruses, and makes available new molecular tools to improve surveys of brassica-infecting viruses.     

Molecular characterization of Pseudomonas syringae isolates from fruit trees and raspberry in Serbia

Infection of fruit trees by Pseudomonas syringae is a potentially serious problem that may limit the establishment and sustained productivity of pome and stone fruit orchards in Serbia. To estimate possible diversity of Pseudomonas syringae fruit trees strains, we collected a set of strains in several areas of Serbia. The samples were taken from infected orchards with raspberry, plum, cherry, sour cherry, peach, pear and apple trees. Genetic diversity of P. syringae strains isolated from fruit trees was determined by using SpeI macrorestriction analysis of genomic DNAs by pulsed-field gel electrophoresis (PFGE) and REP-PCR. Molecular analysis showed that most of isolates had unique profiles, with the exception of isolates from plum and cherry that displayed profiles identical to each other and similar to P. syringae pv. morsprunorum. The study presented here clearly demonstrates the discriminative power of molecular techniques in enabling a detailed analysis of the genetic variations between strains of P. syringae from different pome and stone fruit hosts in Serbia.     

The role of weeds in the epidemiology of pospiviroids

Many weeds that are closely associated with horticultural activities are known as natural reservoirs of plant viruses. However, whether these weeds can also serve as hosts of pospiviroids is not well known. Pospiviroids are naked, non‐coding RNA pathogens that cause severe economic damage in many solanaceous crops. In this study, we examined the overall risk of pospiviroid spreading from weeds to economically important crops, by combining the results from previous inoculation studies with new results coming from a survey, a contact experiment and an inoculation experiment. A survey of commercial ornamental glasshouses revealed that ornamental plants mainly belonging to the Solanaceae harbour pospiviroids, in contrast to weed species sampled in the same places. No new weed hosts could be identified after testing weeds that grew in contact with Tomato apical stunt viroid (TASVd)‐infected plants of tomato (Solanum lycopersicum) and jasmine nightshade (Solanum jasminoides) in an experimental glasshouse. Finally, in mechanical inoculation experiments with TASVd, none of the six tested weed species were determined to be a host at 6 weeks after inoculation. Commonly occurring weed species therefore do not appear to play a significant role as reservoir hosts for pospiviroids. This does not rule out other potential weed hosts that have not yet been tested. Inoculation studies should include rigorous experimental protocols with a sufficient number of replicated as well as adequate positive controls. The information gained through this study may prove useful in future risk assessments for the pospiviroid group.     

Insights into the genetic diversity and evolution of Little cherry virus 1

Little cherry virus 1 (LChV‐1), a member of the recently proposed genus Velarivirus, is a sweet cherry pathogen that has been recently reported to infect other Prunus species and is associated with various plant disorders. In this work the incidence of the virus on its putative hosts and possible mechanisms driving its evolution were investigated. Due to problems encountered with LChV‐1 detection, a new nested RT‐PCR assay was developed and applied. The virus was found to be prevalent in cherry plantations in Greece and only occasionally detected in other Prunus species. Sequences corresponding to the partial RNA‐dependent RNA polymerase (RdRp), heat‐shock protein homologue (HSP70h) and coat protein (CP) genes were determined from Greek LChV‐1 isolates originating from different hosts; these were analysed, along with published homologous genomic regions from other isolates. Phylogenetic analysis of the three genes revealed the segregation of four evolutionary distinct groups showing no host or geography‐based clustering. Mean genetic distances among the four groups were high with the CP region showing the highest divergence, although intragroup variability levels were low. Nevertheless, estimations of the mean ratio of nonsynonymous substitutions per synonymous site to synonymous substitutions per synonymous site (dN/dS) for the partial RdRp, HSP70h and CP indicated that these genomic regions are under negative selection pressure. Interestingly, a recombination event was identified at the 3′ end of RdRp on a Greek virus isolate, thus highlighting the role of this mechanism in the evolutionary history of LChV‐1.     

Evidence for an expanded host range ofFusarium oxysporum f.sp.raphani

The pathogenicity of four isolates ofFusarium oxysporum obtained from infected cultivated rocket (Eruca vesicaria) and wild (sand) rocket (Diplotaxis tenuifolia) was tested on the following cruciferous hosts: stock, radish, wild and cultivated rockets, and various species in the cabbage tribe: cabbage (Brassica oleracea var.sabauda), cauliflower (Brassica oleracea var.botrytis), Brussels sprouts (Brassica oleracea var.gemmifera), broccoli (Brassica oleracea var.italica), turnip (Brassica rapa var.rapa). The results indicated that isolates ofF. oxysporum from cultivated and wild rocket belong to theforma specialis raphani. The isolates from rocket were pathogenic on cabbage, Brussels sprouts, broccoli, turnip, radish and stock; isolates ofF. oxysporum conglutinans from cabbage and radish, and the isolate ofF. oxysporum f.sp.raphani from rape obtained from the ATCC collection, were pathogenic on both cultivated and wild rocket.     

Identification and Partial Characterisation of <Emphasis Type="Italic">Lettuce big-vein associated virus</Emphasis> and <Emphasis Type="Italic">Mirafiori lettuce big-vein virus</Emphasis> in Common Weeds Found Amongst Spanish Lettuce Crops and their Role in Lettuce Big-vein Disease Transmission

The potential role of 10 frequently occurring weed species found amongst Spanish lettuce crops as host plants for the two viruses associated with the lettuce big-vein disease, Lettuce big-vein associated virus (LBVaV) and Mirafiori lettuce big-vein virus (MLBVV), was studied. The results showed that both viruses can infect naturally growing Sonchus oleraceus (common sowthistle) plants, the unique susceptible species detected among the analysed weeds. The sequences of the coat protein (CP) genes of the LBVaV and MLBVV isolates recovered from S. oleraceus plants were determined. Phylogenetic studies revealed a very close relationship between the CP sequences from these weed isolates and those from Spanish lettuce. Moreover, we showed that S. oleraceus can act as a source of lettuce infection by means of Olpidium brassicae, the vector fungus of both viruses.     

Characterization of the first two viruses described from wild populations of hammer orchids (Drakaea spp.) in Australia

Sequences representing the genomes of two distinct virus isolates infecting wild plants of two members of the genus Drakaea (hammer orchids) in Western Australia are described. The virus isolated from Drakaea livida has a bipartite genome of 4490 nt (RNA1) and 2905 nt (RNA2) that shares closest sequence and structural similarity to members of the genus Pecluvirus, family Virgaviridae, described from legumes in the Indian subcontinent and West Africa. However, it differs from pecluviruses by lacking a P39 protein on RNA2 and having a cysteine‐rich protein gene located 3′ of the triple gene block protein genes. It is the first peclu‐like virus to be described from Australia. The name Drakaea virus A is proposed (DVA; proposed member of the family Virgaviridae, genus unassigned). The second virus isolate was identified from Drakaea elastica, a species classed as endangered under conservation legislation. The genome sequence of this virus shares closest identity with isolates of Donkey orchid symptomless virus (DOSV; proposed member of the order Tymovirales, family and genus unassigned), a species described previously from wild Caladenia and Diuris orchids in the same region. These viruses are the first to be isolated from wild Drakaea populations and are proposed to have an ancient association with their orchid hosts.