Sequence diversity within the three agents of groundnut rosette disease

ABSTRACT Sequence diversity was examined in the coat protein (CP) gene of Groundnut rosette assistor virus (GRAV), the overlapping open reading frames (ORFs) 3 and 4 of Groundnut rosette virus (GRV), and the satellite RNA (sat-RNA) of GRV obtained from field isolates from Malawi and Nigeria. These three agents cause groundnut rosette disease, a major disease of groundnut in sub-Saharan Africa (SSA). Sequence analysis showed that the GRAV CP gene was highly conserved (97 to 99%) independent of its geographic source. The nucleotide sequence of the overlapping ORFs 3 and 4 of GRV was highly conserved (98 to 100%) from isolates within a geographic region but less conserved (88 to 89%) between isolates from the two distinct geographic regions. Phylogenetic analysis of the overlapping ORFs 3 and 4 show that the GRV isolates cluster according to the geographic region from which they were isolated, indicating that Malawian GRV isolates are distinct from Nigerian GRV isolates. Similarity within the sat-RNA sequences analyzed ranged from 88 to 99%. Phylogenetic analysis also showed clustering within the sat-RNA isolates according to country of origin, as well as within isolates from two distinct regions of Malawi. Because the GRAV CP sequence is highly conserved, independent of the geographic source of the GRAV isolates, the GRAV CP sequence represents the most likely candidate to use for pathogen-derived resistance in groundnut and may provide effective protection against groundnut rosette disease throughout SSA.     

Transmission biology of Raspberry latent virus, the first aphid-borne reovirus

Raspberry latent virus (RpLV) is a newly characterized reovirus found in commercial raspberry fields in the Pacific Northwest (PNW). Thus far, all members of the plant reoviruses are transmitted in a replicative, persistent manner by several species of leafhoppers or planthoppers. After several failed attempts to transmit RpLV using leafhoppers, the large raspberry aphid, commonly found in the PNW, was tested as a vector of the virus. The virus was transmitted to new, healthy raspberry plants when inoculated with groups of at least 50 viruliferous aphids, suggesting that aphids are vectors of RpLV, albeit inefficient ones. Using absolute and relative quantification methods, it was shown that the virus titer in aphids continued to increase after the acquisition period even when aphids were serially transferred onto fresh, healthy plants on a daily basis. Transmission experiments determined that RpLV has a 6-day latent period in the aphid before it becomes transmissible; however, it was not transmitted transovarially to the next generation. To our knowledge, this is the first report of a plant reovirus transmitted by an aphid. Phylogenetic analyses showed that RpLV is related most closely to but distinct from Rice ragged stunt virus (RRSV), the type member of the genus Oryzavirus. Moreover, the conserved nucleotide termini of the genomic segments of RpLV did not match those of RRSV or other plant reoviruses, allowing us to suggest that RpLV is probably the type member of a new genus in the Reoviridae comprising aphid-transmitted reoviruses.     

蚜虫传播非持久性病毒的取食行为调控机制

蚜虫能够传播上百种植物病毒,是最重要的农业介体昆虫之一。蚜虫在刺探和取食植物过程中,唾液组分会连同附着在口针中的病毒粒子一同被分泌进入植物内,在调节植物诱导抗性、病毒侵染扩散、介体昆虫行为等过程中均有重要作用。本文围绕蚜虫传播病毒和获取病毒2个关键过程,总结分析了蚜虫独特的刺吸取食行为与传毒效率和获毒效率之间的联系;针对取食活动中关键的唾液蛋白在调控植物免疫抗性、帮助病毒侵染过程中的功能,阐述了蚜虫高效传播非持久病毒的分子基础;针对蚜虫的获毒过程,综述了病毒侵染植物间接调控蚜虫趋向和行为的作用方式。这些研究的开展将为解释蚜虫和病毒协同侵染的分子机制以及有效开展基于蚜虫取食行为调控的病虫害防控新技术提供思路。     

Aphid stylet activities during potyvirus acquisition from plants and anin vitro system that correlate with subsequent transmission

The behavioural events associated with acquisition of tobacco etch potyvirus by starvedMyzus persicae during single, electrically-recorded penetrations of plants or a Parafilm membrane were compared. Twenty nine percent of aphids acquired virus from plants and subsequently transmitted to test plants. Stylet puncture of the plasmalemma, indicated by a potential drop (pd) to the intracellular signal voltage level, occurred during 84% of penetrations, and virus transmission was always associated with this behavioural event during acquisition. Periods of intracellular stylet tip location, known as pd phase II, ranged from 3.6–12.2s, and always comprised three consecutive sub-phases, designated II1, II2 and II3. Ninety six percent of pds included distinct pulses during phase II3. A waveform which closely resembled these pulses was produced by 59% of aphids that probed a virus suspension through a Parafilm membrane; nineteen percent of the aphids subsequently transmitted membrane-acquired virus and transmission was significantly associated with the occurrence of the phase II3-like pulses during acquisition. The duration of occurrence of recorded phase II3 pulses, either on plants or thein vitro system, did not influence the virus transmission efficiency of aphids. The association of virus uptake from aqueous suspension with a particular behavioural activity is discussed as evidence for the ingestion-egestion hypothesis for nonpersistent transmission. Starved aphids acquiring virus from infected leaf tissue or thein vitro system had significantly higher transmission efficiencies than non-starved aphids. Starved and non-starved insects were electrically-recorded penetrating the artificial membrane, and again there was a clear difference in transmission efficiency (starved aphids, 26%; non-starved aphids, 2%). The higher transmission efficiency of starved insects could not be explained by behavioural differences, and the results lend support to the hypothesis that non-behavioural factors determine the enhancement of potyvirus transmission by preacquisition starvation.Abbreviations BMV Beet mosaic virus - EMF Electromotive force - HAT Highly aphid-transmissible - HC Helper component - pd Potential drop - PVY Potato virus Y - TEV Tobacco etch virus     

The effect of potato leafroll virus on the biology of Myzus persicae

Since potato leafroll virus multiplies in the green peach aphid,Myzus persicae, the effect of the virus on the biology of its vector was investigated. Observations were made regarding the longevity and the reproduction rate of viruliferous and non-viruliferous aphids on leafroll-diseased and healthy plants ofPhysalis floridana. The same matters were investigated for both viruliferous and non-viruliferous aphids on seedlings of Chinese cabbage (Brassica pekinensis). It was shown that on leafroll-diseased plants ofP. floridana the aphids produced more progeny than on healthy ones, although the average number of progeny produced per day in both cases was almost the same. On healthy Chinese cabbage seedlings there was no difference in average length of the larval and adult stages, number of progeny per aphid, and number of progeny per day, between viruliferous and non-viruliferous aphids. Evidence was obtained that the virus does not influence the development of its vector. Measurements of oxygen consumption of both viruliferous and non-viruliferous aphids point in the same direction.     

Transmission of barley yellow dwarf virus by cereal aphids collected from different habitats on cereal farms

Cereal aphids were collected from cereal crops, from Poa annua within cereal fields, from Lolium perenne pastures and from wild grasses in hedge bottoms and around farm buildings. The frequency of barley yellow dwarf virus (BYDV) transmission was assessed by aphid transmission tests. There were differences in transmission rates between aphid species, between host species and between years. The transmission rates of Rhopalosiphum padi from the different host species were broadly similar whereas for Sitobion avenae, P. annua within cereal fields was significantly better than the other host species. Wild grasses other than P. annua were relatively poor sources of virus. A large percentage of aphids frequently transmitted more than one strain, suggesting that host plants are often infected with more than one BYDV strain.     

Stability of the aphid transmission phenotype in cucumber mosaic virus

The stability of the aphid transmission phenotype in seven field isolates of cucumber mosaic virus (CMV) was studied, using aphids Aphis gossypii and Myzus persicae . Field isolates, obtained from four vegetable crops, were propagated in squash and Nicotiana glutinosa , and passaged by either aphid transmission or mechanical transfer. All seven isolates were transmissible by both aphids and this aphid transmission phenotype was stable after 20–24 mechanical passages. Upon further mechanical passaging, one of the seven isolates, CMV-2 A1-MT 60x, lost its transmissibility by Myzus persicae but was still transmissible by Aphis gossypii , although at a reduced rate. Isolates maintained by both aphid transmission and mechanical transfer were transmitted more efficiently by Aphis gossypii than by Myzus persicae . A comparison of the RNA profiles showed no major differences among the CMV isolates before and after mechanical passage.     

Divergent effects of PVY-infected potato plant on aphids

Host plant selection by aphids can be positively or negatively affected when plants are infected by phytoviruses. Potato plants infected by Potato virus Y (PVY), a non-persistent virus, are reported to affect settling behaviour and growth parameters of Myzus persicae Sulzer and Macrosiphum euphorbiae Thomas. Using the Electrical penetration graph system (EPG), we demonstrated that PVY-infection of potato plants influences the feeding behaviour of these two aphid species. Myzus persicae exhibited increased phloem sap ingestion and reduced non-probing duration. Macrosiphum euphorbiae showed delayed stylet insertion, reduced activity in the phloem vessels and an enhanced non-probing duration. In addition, we showed that these two species exhibited different transmission rates. The opposite effects of PVY-infected potato plant on these two aphids are discussed in terms of PVY spreading in the field.     

The genus <Emphasis Type="Italic">Luteovirus</Emphasis> from infection to disease

Luteoviruses are economically important plant viruses. Specifically, barley yellow dwarf virus is epiphytotic to almost all small-grain cereal growing areas. The disease cycle is complex. This luteovirus has evolved several intelligent mechanisms to communicate with both plant and phloem-feeding insect-vector aphid. Environmental cues influence disease severity, aphid infestation and viral load. Within an aphid, virus circulates persistently in a non-propagative manner and is transmitted selectively to the host plants. Selection of viruses within aphids has a role in virus isolate prevalence over a specific area. In the host-plant system, the virus has to release its single sense-strand RNA genome (approx. 5.6 to 6 kb), translate and subsequently replicate its genome using its own replicase and host machinery. This review summarizes our current understanding of disease epidemiology and reviews the current literature encompassing viral infectivity, economic impact and control measures.     

Evidence of no horizontal or vertical transmission of tomato severe rugose virus by Bemisia tabaci MEAM1

Bemisia tabaci is important in agriculture worldwide, mainly because it is a vector of numerous plant viruses, probably the most important of which are members of the genus Begomovirus. Dozens of begomoviruses have been reported to infect tomato plants in Brazil, although tomato severe rugose virus (ToSRV) predominates in tomato crops. ToSRV, found so far only in Brazil, is efficiently transmitted by B. tabaci MEAM1. However, no studies have assessed the occurrence of vertical and horizontal transmission of the virus in the insect, which may have epidemiological consequences affecting disease management. This study evaluated the possibility of transmission of ToSRV between whiteflies during copulation and transovarial transmission from viruliferous females of B. tabaci MEAM1 to their progeny. Transmission of ToSRV did not occur during mating between males and females of B. tabaci MEAM1. Aviruliferous males and females confined with viruliferous insects of the opposite sex were also unable to transmit the virus to tomato plants. ToSRV was detected, by PCR, in the ovaries of viruliferous females of B. tabaci MEAM1 but not in eggs, nymphs, or adults of the progeny of viruliferous females. Adult progeny of viruliferous females also did not transmit ToSRV to tomato plants. Together, the results indicate that vertical and horizontal transmission of ToSRV by B. tabaci MEAM1 is unlikely. Sustainable management of the tomato golden mosaic disease caused by ToSRV should continue to focus on using resistant varieties, managing sources of inoculum around tomato fields, and rational chemical control of the vector.     

一种蚜虫持久性传病的长豇豆黄化型病毒

 1983年6月,在南京郊区的长豇豆上采到1株表现植株矮缩症状的C-7病毒分离物。接种试验证明,它不能摩擦接种传病,但可以由豆蚜(Aphis craccivora)、棉蚜(A.gossypii)和桃蚜(Myzus persicae)以持久性方式传病。寄主范围测定的结果表明:分离物可以侵染长豇豆、豇豆、蚕豆、大豆、菜豆、豌豆、赤豆、利马豆、苜蓿、红三叶、地三叶、绛三叶、葫芦巴,紫云英和苕子等15种豆科植物和曼陀罗1种茄科植物。这些植物大都出现植株矮化,叶片扭曲,卷缩或僵缩,不能开花结实等症状。豆蚜的传病性状中,获毒饲育的最短传病时间为3小时,接毒饲育最短传病时间为10分钟,循回期是24小时左右。但是,传病率最高的获毒饲育时间是2~3天,接毒饲育时间在1天以上。接种1头蚜虫就具有传病能力,5头蚜虫能达到100%的传病率。蚜虫可以终身传毒,蜕皮不影响其传毒力,但传毒有间歇性。根据它的基本性状,病毒C-7分离物是一种豆科植物的黄化型病毒,可能是属于大麦黄矮病毒组(Luteovirus Group)的成员。     

台湾75毛豆病毒病防治技术研究

研究表明,台湾75毛豆病毒病主要由蚜虫传毒而致,其发生与蚜虫的成虫迁飞时间与发生量关系极为密切.若成虫高峰正遇毛豆1、2叶展平期,毛豆上落蚜量大,发生就重.所以早播的重于迟播的,露地栽培重于套播的.防治上应以栽培避蚜为主,药剂防治为辅,才可取得理想效果.     

刺槐—花生矮化病毒的初侵染源

 1988~1991年调查河北、河南、山东和北京地区刺槐花叶病树率为4~87.5%,一般30%左右。蚜传试验说明:豆蚜(Aphis craccivora)能将花生矮化病毒(PSV)从刺槐传播到花生。在田间经常观察到PSV由刺槐花叶病树林向花生传播,形成病害发生梯度。1990~1991两年河南开封田间系统观察,5月中旬至6月上、中旬,蚜虫在刺槐上形成繁殖高峰并产生有翅蚜虫向花生地迁飞,随后花生地内出现PSV病株。1990和1991两年河北滦县、迁安病区调查,未发现PSV种传苗,开封PSV感染病株种子,PSV种传率0.025%。PSV引起花生病害一般年份发病很轻。与种传相比,刺槐作为病害初侵染源,对PSV在花生上流行起更为重要的作用。     

Use of insect antifeedants against aphid vectors of plant virus disease

The probing and feeding behaviour of aphids can result in uptake of viruses from infected plants and subsequent transmission to healthy plants. It is possible to interfere with virus acquisition and transmission by influencing aphid host-selection and feeding behaviour with antifeedant chemicals. Published work is reviewed and new work is presented in this paper. The two most successful classes of antifeedants against aphids are (a) compounds derived from the aphid alarm pheromone and (b) plant-derived antifeedants such as the sesquiterpene (-)-polygodial. Results with these and other compounds are discussed in terms of antifeedant activity against Myzus persicae, their effects on resistant aphids and the evidence for dec?ease in virus spread by aphids in laboratory and field.     

The recombinant isolate of cucurbit aphid-borne yellows virus from Brazil is a polerovirus transmitted by whiteflies

The severe yellowing disease (amarelão) on melon plants is a serious problem in Brazil, although the causative agent remained unknown for a long time. Recently, recombinant isolates of cucurbit aphid-borne yellows virus (CABYV) were reported as the possible causative agents of this disease on melon plants. Although aphids are known to be the vectors of the common type of CABYV isolates, almost no aphid colony was observed in the major melon fields in Brazil with high incidence of the severe yellowing disease. In contrast, whiteflies are often abundant. Based on this observation, the hypothesis of the transmission of recombinant CABYV by whiteflies was evaluated. After thorough transmission experiments, we found that this recombinant CABYV isolate was transmitted by the whitefly Bemisia tabaci MEAM1, but not by Aphis gossipii. Furthermore, the host response by whitefly-based inoculation in cucurbits and other indicator plants showed differences in host range when compared to the common type of CABYV. Due to its transmissibility by the whitefly and the distant relationship of the P3/P5 protein to CABYV, the name “cucurbit whitefly-borne yellows virus” is proposed for this recombinant CABYV. This is the second report of polerovirus transmission by the whitefly B. tabaci, following the report of pepper whitefly-borne vein yellows virus.     

Effect of temperature, vector life stage, and plant access period on transmission of banana bunchy top virus to banana

The study of the transmission biology of insect-borne plant viruses is important to develop disease control practices. We characterized the transmission of a nanovirus, Banana bunchy top virus (BBTV), by its aphid vector Pentalonia nigronervosa Coquerel (Hemiptera, Aphididae) with respect to temperature, vector life stage, and plant access time. Adult aphids transmitted BBTV more efficiently than third instar nymphs at all temperatures tested. Adult aphids transmitted the virus more efficiently at 25 and 30 degrees C than at 20 degrees C, but temperature had no impact on transmission efficiency by nymphs. By decoupling the relationship between temperature and aphid BBTV acquisition or inoculation, we determined that temperature affected inoculation events more strongly than acquisition. Longer plant access periods increased viral acquisition and inoculation efficiencies in a range of 60 min to 24 h. Both BBTV acquisition and inoculation efficiencies peaked after 18 h of plant access period. We also show that BBTV transmission by P. nigronervosa requires a latent period. Our results demonstrate that vector transmission of BBTV is affected by temperature, vector life stage, and plant access period.     

Effects of Susceptibility of Test Plants on Modes of Cucumovirus Transmission by Myzus persicae

In sequential transmission tests of Peanut stunt virus S (PSV-S) and Cucumber mosaic virus V (CMV-V) using Myzus persicae, these viruses behave as a semipersistent virus in Phaseolus vulgaris, but as a nonpersistent virus in Nicotiana tabacum, regardless of the species of plant used as the virus source. In addition, viruliferous aphids retained virus infectivity and transmitted it to P. vulgaris, even after they lost infectivity to N. tabacum. Apparently, the mode of transmission by the aphids differs depending on the plant species used for the assay. After mechanical inoculation with purified PSV-S or CMV-V, P. vulgaris appeared more susceptible to PSV-S than N. tabacum. However, the susceptibility to CMV-V appeared similar in both assay plants. Received 22 May 2000/ Accepted in revised form 14 September 2000     

Flonicamid, a novel insecticide with a rapid inhibitory effect on aphid feeding

Flonicamid (IKI220; N-cyanomethyl-4-trifluoromethylnicotinamide), a pyridinecarboxamide compound, is a novel systemic insecticide with selective activity against hemipterous pests, such as aphids and whiteflies, and thysanopterous pests. The purpose of this study is to clarify the biological properties of flonicamid against aphids. Flonicamid is very active against aphids, regardless of differences in species, stages and morphs. This compound inhibited the feeding behaviour of aphids within 0.5 h of treatment without noticeable poisoning symptoms such as convulsion, and this antifeeding activity was not recoverable until death. The nymphs born from adults exposed to flonicamid for 3 h showed high mortality. The effect of flonicamid on the feeding activity of an individual aphid was studied using electronic monitoring of insect feeding behaviour (EMIF). Although the treated aphid attached the head of its proboscis to the leaf surface, salivation and sap feeding were strongly inhibited. These results suggest that the main insecticidal mechanism of flonicamid is starvation based on the inhibition of stylet penetration to plant tissues.     

Transmission efficiency of Papaya ringspot virus by three aphid species

The transmission efficiency of Papaya ringspot virus (PRSV) by three aphid vectors (i.e., Aphis gossypii, A. craccivora, and Myzus persicae) was studied. Efficiency was measured by single-aphid inoculation, group inoculation (using five aphids), duration of virus retention, and the number of plants following a single acquisition access period (AAP) to which the aphids could successfully transmit the virus. Single-aphid inoculation studies indicated that M. persicae (56%) and A. gossypii (53%) were significantly more efficient in transmitting PRSV than A. craccivora (38%). Further, in the former two species, the time required for initiation of the first probe on the inoculation test plant was significantly shorter compared to A. craccivora. PRSV transmission efficiency was 100% in all three species when a group of five aphids were used per plant. There was a perceptible decline in transmission efficiency as the sequestration period increased, although M. persicae successfully transmitted PRSV after 30 min of sequestration. A simple leaf-disk assay technique was employed for evaluating the transmission efficiency of three species of aphids. The results of leaf-disk assays also indicated that A. gossypii (48%) and M. persicae (56%) were more efficient PRSV vectors than A. craccivora. Using leaf-disk assays, the ability of individual aphids to inoculate PRSV serially to a number of plants was studied. Following a single AAP on an infected leaf, M. persicae was more efficient than the other two species with 52.5% transmission after the first inoculation access period (IAP). However, its inoculation efficiency significantly decreased with the second and subsequent IAPs. A. gossypii was able to transmit PRSV sequentially up to four successive leaf disks, but with significantly declining efficiency. Since A. gossypii is reported to be the numerically dominant vector in south India in addition to being a more efficient vector capable of inoculating PRSV to multiple plants, it should be the target vector for control strategies.     

一种由麦二叉蚜和麦长管蚜传播的小麦黄矮病毒株系的鉴定

 通过对采自我国内蒙古丰镇县13个春小麦病株标样进行4种蚜虫连续传毒比较,不同温度饲毒和接种试验,薄膜饲毒,2种蚜虫成若蚜传毒比较和血清学酶标试验等,明确我国内蒙古丰镇县小麦黄矮病毒株系为由麦二叉蚜和麦长管蚜有效传播的麦二叉蚜麦长管蚜株系(GAV)。对于这个株系,禾缢管蚜与玉米蚜是不能传播的。这种株系在我国冬麦区春麦区都有分布。