Acyrthosiphon Kondoi Research Articles

Discovery of insecticide resistance in field-collected populations of the aphid pest, Acyrthosiphon kondoi Shinji.

The bluegreen aphid (Acyrthosiphon kondoi) is a worldwide pest of alfalfa, pulses, and other legume crops. An overreliance on insecticides to control A. kondoi has potentially placed populations under selection pressure favouring resistant phenotypes, but to date, there have been no documented cases of insecticide resistance. Recently, Australian growers began reporting that conventional insecticides were failingto adequately control A. kondoi populations, prompting this laboratory-based investigation into whether these populations have evolved resistance. We discovered four A. kondoi populations with moderate resistance (10-40-fold) to three different insecticide groups: organophosphates, carbamates and pyrethroids. However, A. kondoi populations showed no resistance to the butenolide, flupyradifurone. We were unable to identify general metabolic mechanisms using synergist assays (cytochromes P450, glutathione S-transferases, or esterases), indicating that further detailed molecular investigations to characterise the putative resistance mechanism are needed. Insecticide-resistant A. kondoi present an emerging challenge to Australian agriculture. Growers require new tools and updated strategies, including access to newer chemistries, to alleviate their reliance on the few insecticides currently registered against A. kondoi. The implications of insecticide resistant A. kondoi for future management, the potential mechanisms of resistance, and future research priorities are discussed. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Ethylene Is Not Essential for R-Gene Mediated Resistance but Negatively Regulates Moderate Resistance to Some Aphids in Medicago truncatula.

Ethylene is important for plant responses to environmental factors. However, little is known about its role in aphid resistance. Several types of genetic resistance against multiple aphid species, including both moderate and strong resistance mediated by R genes, have been identified in Medicago truncatula. To investigate the potential role of ethylene, a M. truncatula ethylene- insensitive mutant, sickle, was analysed. The sickle mutant occurs in the accession A17 that has moderate resistance to Acyrthosiphon kondoi, A. pisum and Therioaphis trifolii. The sickle mutant resulted in increased antibiosis-mediated resistance against A. kondoi and T. trifolii but had no effect on A. pisum. When sickle was introduced into a genetic background carrying resistance genes, AKR (A. kondoi resistance), APR (A. pisum resistance) and TTR (T. trifolii resistance), it had no effect on the strong aphid resistance mediated by these genes, suggesting that ethylene signaling is not essential for their function. Interestingly, for the moderate aphid resistant accession, the sickle mutant delayed leaf senescence following aphid infestation and reduced the plant biomass losses caused by both A. kondoi and T. trifolii. These results suggest manipulation of the ethylene signaling pathway could provide aphid resistance and enhance plant tolerance against aphid feeding.

An RNAi supplemented diet as a reverse genetics tool to control bluegreen aphid, a major pest of legumes

Aphids are important agricultural pests causing major yield losses worldwide. Since aphids can rapidly develop resistance to chemical insecticides there is an urgent need to find alternative aphid pest management strategies. Despite the economic importance of bluegreen aphid (Acyrthosiphon kondoi), very few genetic resources are available to expand our current understanding and help find viable control solutions. An artificial diet is a desirable non-invasive tool to enable the functional characterisation of genes in bluegreen aphid and discover candidate target genes for future use in RNA interference (RNAi) mediated crop protection against aphids. To date no artificial diet has been developed for bluegreen aphid, so we set out to develop a suitable diet by testing and optimising existing diets. Here, we describe an artificial diet for rearing bluegreen aphid and also provide a proof of concept for the supplementation of the diet with RNAi molecules targeting the salivary gland transcript C002 and gap gene hunchback, resulting in bluegreen aphid mortality which has not yet been documented in this species. Managing this pest, for example via RNAi delivery through artificial feeding will be a major improvement to test bluegreen aphid candidate target genes for future pest control and gain significant insights into bluegreen aphid gene function.

Obligate bacterial endosymbionts limit thermal tolerance of insect host species

The thermal tolerance of an organism limits its ecological and geographic ranges and is potentially affected by dependence on temperature-sensitive symbiotic partners. Aphid species vary widely in heat sensitivity, but almost all aphids are dependent on the nutrient-provisioning intracellular bacterium Buchnera, which has evolved with aphids for 100 million years and which has a reduced genome potentially limiting heat tolerance. We addressed whether heat sensitivity of Buchnera underlies variation in thermal tolerance among 5 aphid species. We measured how heat exposure of juvenile aphids affects later survival, maturation time, and fecundity. At one extreme, heat exposure of Aphis gossypii enhanced fecundity and had no effect on the Buchnera titer. In contrast, heat suppressed Buchnera populations in Aphis fabae, which suffered elevated mortality, delayed development and reduced fecundity. Likewise, in Acyrthosiphon kondoi and Acyrthosiphon pisum, heat caused rapid declines in Buchnera numbers, as well as reduced survivorship, development rate, and fecundity. Fecundity following heat exposure is severely decreased by a Buchnera mutation that suppresses the transcriptional response of a gene encoding a small heat shock protein. Similarly, absence of this Buchnera heat shock gene may explain the heat sensitivity of Ap. fabae Fluorescent in situ hybridization revealed heat-induced deformation and shrinkage of bacteriocytes in heat-sensitive species but not in heat-tolerant species. Sensitive and tolerant species also differed in numbers and transcriptional responses of heat shock genes. These results show that shifts in Buchnera heat sensitivity contribute to host variation in heat tolerance.

Gene pyramids and the balancing act of keeping pests at bay.

This article comments on: Kamphuis LG, Klingler JP, Jacques S, Gao L-l, Edwards OR, Singh KB. 2019. Additive and epistatic interactions between AKR and AIN loci conferring bluegreen aphid resistance and hypersensitivity in Medicago truncatula. Journal of Experimental Botany 70, 4887-4902.

Additive and epistatic interactions between AKR and AIN loci conferring bluegreen aphid resistance and hypersensitivity in Medicago truncatula.

Aphids, including the bluegreen aphid (BGA; Acyrthosiphon kondoi), are important pests in agriculture. Two BGA resistance genes have been identified in the model legume Medicago truncatula, namely AKR (Acyrthosiphon kondoi resistance) and AIN (Acyrthosiphon induced necrosis). In this study, progeny derived from a cross between a resistant accession named Jester and a highly susceptible accession named A20 were used to study the interaction between the AKR and AIN loci with respect to BGA performance and plant response to BGA infestation. These studies demonstrated that AKR and AIN have additive effects on the BGA resistance phenotype. However, AKR exerts dominant suppression epistasis on AIN-controlled macroscopic necrotic lesions. Nevertheless, both AKR and AIN condition production of H2O2 at the BGA feeding site. Electrical penetration graph analysis demonstrated that AKR prevents phloem sap ingestion, irrespective of the presence of AIN. Similarly, the jasmonic acid defense signaling pathway is recruited by AKR, irrespective of AIN. This research identifies an enhancement of aphid resistance through gene stacking, and insights into the interaction of distinct resistance genes against insect pests.

Insecticidal Activity of Diverse Extracts of the Citrullus colocynthis (L.) Against the Blue Green Aphid Acyrthosiphon Kondoi Shinji

The efficiency of three different extracts from stem and leaf of Citrullus colocynthis against the blue green aphid Acyrthosiphon kondoi Shinji was studied. Results showed that the ethanolic extracts from the two parts of C.colocynthis caused higher mortalities of aphids after 48 hours (96 % and 93 % for the stem and leaf, respectively). Toxicological parameters demonstrated that ethanolic extract from the stem of C.colocynthis was more toxic to aphids than leaf extract. This study suggests that detailed chemistry on the stem of C. colocynthis is required before applying it in the integrated pest management of aphids.

Establishing alighting preferences and species transmission differences for Pea seed-borne mosaic virus aphid vectors

Pea seed-borne mosaic virus (PSbMV) infection causes a serious disease of field pea (Pisum sativum) crops worldwide. The PSbMV transmission efficiencies of five aphid species previously found landing in south-west Australian pea crops in which PSbMV was spreading were studied. With plants of susceptible pea cv. Kaspa, the transmission efficiencies of Aphis craccivora, Myzus persicae, Acyrthosiphon kondoi and Rhopalosiphum padi were 27%, 26%, 6% and 3%, respectively. Lipaphis erysimi did not transmit PSbMV in these experiments. The transmission efficiencies found for M. persicae and A. craccivora resembled earlier findings, but PSbMV vector transmission efficiency data were unavailable for A. kondoi, R. padi and L. erysimi. With plants of partially PSbMV resistant pea cv. PBA Twilight, transmission efficiencies of M. persicae, A. craccivora and R. padi were 16%, 12% and 1%, respectively, reflecting putative partial resistance to aphid inoculation. To examine aphid alighting preferences over time, free-choice assays were conducted with two aphid species representing efficient (M. persicae) and inefficient (R. padi) vector species. For this, alatae were set free on multiple occasions (10–15 repetitions each) amongst PSbMV-infected and mock-inoculated pea or faba bean (Vicia faba) plants. Following release, non-viruliferous R. padi alatae exhibited a general preference for PSbMV-infected pea and faba bean plants after 30min–4h, but preferred mock-inoculated plants after 24h. In contrast, non-viruliferous M. persicae alatae alighted on mock-inoculated pea plants preferentially for up to 48h following their release. With faba bean, M. persicae preferred infected plants at the front of assay cages, but mock-inoculated ones their backs, apparently due to increased levels of natural light there. When preliminary analyses were performed to detect PSbMV-induced changes in the volatile organic compound profiles of pea and faba bean plants, higher numbers of volatiles representing a range of compound groups (such as aldehydes, ketones and esters) were found in the headspaces of PSbMV-infected than of mock-inoculated pea or faba bean plants. This indicates PSbMV induces physiological changes in these hosts which manifest as altered volatile emissions. These alterations could be responsible for the differences in alighting preferences. Information from this study enhances understanding of virus-vector relationships in the PSbMV-pea and faba bean pathosystems.

Pea seed-borne mosaic virus Pathosystem Drivers under Mediterranean-Type Climatic Conditions: Deductions from 23 Epidemic Scenarios.

Drivers of Pea seed-borne mosaic virus (PSbMV) epidemics in rainfed field pea crops were examined under autumn to spring growing conditions in a Mediterranean-type environment. To collect aphid occurrence and PSbMV epidemic data under a diverse range of conditions, 23 field pea data collection blocks were set up over a 6-year period (2010 to 2015) at five locations in the southwest Australian grain-growing region. PSbMV infection levels in seed sown (0.1 to 13%), time of sowing (22 May to 22 June), and cultivar (Kaspa or PBA Twilight) varied with location and year. Throughout each growing season, rainfall data were collected, leaf and seed samples were tested to monitor PSbMV incidence in the crop and transmission from harvested seed, and sticky traps were used to monitor flying aphid numbers. Winged migrant Acyrthosiphon kondoi, Lipaphis erysimi, Myzus persicae, and Rhopalosiphum padi were identified in green tile traps in 2014 and 2015. However, no aphid colonization of field pea plants ever occurred in the blocks. The deductions made from collection block data illustrated how the magnitude of PSbMV spread prior to flowering is determined by two primary epidemic drivers: (i) PSbMV infection incidence in the seed sown, which defines the magnitude of virus inoculum source for within-crop spread by aphids, and (ii) presowing rainfall that promotes background vegetation growth which, in turn, drives early-season aphid populations and the time of first arrival of their winged migrants to field pea crops. Likely secondary epidemic drivers included wind-mediated PSbMV plant-to-plant contact transmission and time of sowing. PSbMV incidence at flowering time strongly influenced transmission rate from harvested seed to seedlings. The data collected are well suited for development and validation of a forecasting model that informs a Decision Support System for PSbMV control in field pea crops.

Genetic Mapping of a Major Resistance Gene to Pea Aphid (Acyrthosipon pisum) in the Model Legume Medicago truncatula.

Resistance to the Australian pea aphid (PA; Acyrthosiphon pisum) biotype in cultivar Jester of the model legume Medicago truncatula is mediated by a single dominant gene and is phloem-mediated. The genetic map position for this resistance gene, APR (Acyrthosiphon pisum resistance), is provided and shows that APR maps 39 centiMorgans (cM) distal of the A. kondoi resistance (AKR) locus, which mediates resistance to a closely related species of the same genus bluegreen aphid (A. kondoi). The APR region on chromosome 3 is dense in classical nucleotide binding site leucine-rich repeats (NLRs) and overlaps with the region harbouring the RAP1 gene which confers resistance to a European PA biotype in the accession Jemalong A17. Further screening of a core collection of M. truncatula accessions identified seven lines with strong resistance to PA. Allelism experiments showed that the single dominant resistance to PA in M. truncatula accessions SA10481 and SA1516 are allelic to SA10733, the donor of the APR locus in cultivar Jester. While it remains unclear whether there are multiple PA resistance genes in an R-gene cluster or the resistance loci identified in the other M. truncatula accessions are allelic to APR, the introgression of APR into current M. truncatula cultivars will provide more durable resistance to PA.

An insecticide baseline study of Australian broadacre aphids

Brevicoryne brassicae (Linnaeus), Lipaphis pseudobrassicae (Davis), Acyrthosiphon kondoi (Shinji), Aphis craccivora (Koch) and Rhopalosiphum padi (Linnaeus) are among the most important aphid pests in Australian broadacre systems. In this study a leaf-dip method was used to assay pirimicarb, dimethoate, α-cypermethrin and imidacloprid against field populations collected from Victoria, New South Wales, South Australia, Western Australia and Queensland. This research established toxicity baseline data that will be important for future monitoring of insecticide responses in broadacre crops. It also provided an opportunity to identify any chemical tolerance that may be evolving in these pests. Acyrthosiphon craccivora populations showed differences in their responses to dimethoate, pirimicarb and imidacloprid (but not to α-cypermethrin), indicating possible shifts in field sensitivity to these three chemicals. Rhopalosiphum padi had the lowest sensitivity to all insecticides tested, with two populations (collected from South Australia and Queensland) showing less than 100% mortality when tested at the field rate of α-cypermethrin. There were few differences in insecticide responses between populations of the other three species. Continued screening of A. craccivora and R. padi populations is needed to fully assess the current status of tolerance among field populations and to strengthen resistance management tactics.

Diversity for resistance to a moderately virulent bluegreen aphid (Acyrthosiphon kondoi Shinji) population in Trifolium species

The resistance of a diverse range of Trifolium species (clovers) to a highly virulent bluegreen aphid (BGA, Acyrthosiphon kondoi Shinji) population (Urrbrae 2011) collected in South Australia was assessed in greenhouse and field experiments, with the aims of determining the potential impact of this insect pest on biomass and identifying resistant genotypes for future plant-breeding activities. Resistance to BGA was found in populations of clovers that show some level of outcrossing—white clover (T. repens L.), rose clover (T. hirtum All.), crimson clover (T. incarnatum L.) and red clover (T. pratense L.)—and in one entry of the inbreeding subspecies of subterranean clover, T. subterraneum L. subsp. subterraneum (Katzn. and Morley). Resistance was not found in T. s. brachycalycinum (Katzn. and Morley) or T. s. yanninicum (Katzn. and Morley). In a greenhouse experiment, damage from BGA resulted in forage yield penalties of 72–100% when aphids were inoculated at 14 days after sowing and 13−74% when inoculated at 42 days after sowing, showing that in optimum conditions BGA can be a serious pest of clovers. Observations of severe damage caused by BGA in two regenerating field trials in southern New South Wales confirmed that field damage could occur in seasons favourable to aphid growth and reproduction. The severe damage that BGA can cause to clovers, and the sources of resistance we found, suggest that breeding for BGA resistance in clovers is warranted and feasible.

Insecticide Efficacy and Application Timing for Blue Alfalfa Aphid Control, 2014

The objective of the study was to evaluate the efficacy of the insecticidal compounds via ground spray applications at various times for control of BAA on alfalfa grown for hay production under low desert growing conditions. A field study was conducted during the spring of 2014 at the UC Desert Research and Extension Center. A stand of alfalfa, VAR. ‘CUF-101,’ was used for the experiment. The experimental design was an RCB using four replicates with 10 insecticidal treatments and an untreated check. Plots measured 13.33 ft by 50 ft and insecticide treatments …

Effects of aphid feeding and associated virus injury on grain crops in Australia

We review yield effects caused by aphid feeding and associated virus injury to cereal, oilseed and pulse crops, and estimate the potential economic loss caused by aphids in Australia. Potential yield reduction due to aphids was determined through a survey of quantitative data from experiments that assessed aphids' effect on grain yield. In cereals, four aphids caused damage; on barley, feeding injury caused by Rhopalosiphum padi + Rhopalosiphum maidis was most damaging in terms of yield reduction (25.5%) with an economic loss of $19/ha. Barley yellow dwarf virus transmitted by R. padi + Sitobion miscanthi was more damaging than direct feeding, causing a yield reduction of 39% and economic loss of $21/ha for wheat. On canola, beet western yellow virus transmitted by Myzus persicae caused the highest yield reduction of 34% and economic loss of $115/ha, although this was measured through artificial inoculations. Feeding injury was high in Brevicoryne brassicae which caused an average yield reduction of 34% and associated economic loss of $88.5/ha, while Lipaphis erysimi and M. persicae had negligible economic effects but more data are needed. On pulses, the most economically damaging (unidentified) aphids feeding on lupins caused a yield reduction of 43% and economic loss of $24/ha. The aphids M. persicae + Aphis craccivora + Acyrthosiphon kondoi reduced lupin yields by 13% and economic returns by $7.40/ha. On field peas, a 14% reduction in yield was caused by transmitted viruses such as pea seed-borne mosaic virus which caused economic losses of $20.50/ha. In total, feeding and virus injuries resulted in potential economic costs of $241 and $482 million/year, respectively. Although this review provides estimates of potential yield and economic losses due to aphids, few data were available for some crops, aphid species or regions (e.g. oats). Nevertheless, economic costs associated with aphids appear substantial.

Preference of Aphids (Hemiptera: Aphididae) for Lucerne, Maize, Soybean and Wheat and their Potential as Prospective Border Crops forPotato virus yManagement in seed Potatoes

Aphid landing rates, species composition and abundance on lucerne (Medicago sativa), maize (Zea mays), soybean (Glycine max), wheat (Triticum aestivum) and potato (Solanum tuberosum) were determined in a small-scale field trial to identify potential crops as prospective border plants for seed potatoes to reduce the spread of Potato virus (PVY) in South Africa. Aphids were sampled using green bucket traps to estimate aphid landing rates, whereas leaf counts and sweep-netting were used to identify colonizing species. Of the 34 species or species groups collected with green bucket traps, 18 were previously known vectors of PVY. The most abundant vector species or species groups were Acyrthosiphon pisum, Macrosiphum euphorbiae, Metopolophium dirhodum, Myzus persicae, Rhopalosiphum padi and Sitobion spp. The only other species that occurred in high numbers, but whose vector status is unknown, were Acyrthosiphon kondoi and Tetraneura fusiformis. Landing patterns suggest that A. kondoi, A. pisum and M. dirhodum may be able to discriminate between plant species and select their preferred host plant, if available within a habitat patch, in the pre-alighting phase, whereas other species such as R. padi seemed less selective. Results on aphid landing together with colonization of the crops evaluated suggest that maize and wheat show the highest potential as possible crop border plants in regions where aphids colonizing Poaceae, and lucerne in regions where aphids colonizing Fabaceae are abundant. Soybean appears to be less suitable because cultivars with a high trichome density, which reduces colonization by aphids, are preferentially planted in South Africa.

Genetic improvement of subterranean clover (Trifolium subterraneum L.). 2. Breeding for disease and pest resistance

Subterranean clover (Trifolium subterraneum L.) is the most widely sown pasture legume in southern Australia and resistance to important diseases and pests has been a major plant-breeding objective. Kabatiella caulivora, the cause of clover scorch, is the most important foliar fungal pathogen, and several cultivars have been developed with resistance to both known races. Screening of advanced breeding lines has been conducted to prevent release of cultivars with high susceptibility to other important fungal foliar disease pathogens, including rust (Uromyces trifolii-repentis), powdery mildew (Oidium sp.), cercospora (Cercospora zebrina) and common leaf spot (Pseudopeziza trifolii). Several oomycete and fungal species cause root rots of subterranean clover, including Phytophthora clandestina, Pythium irregulare, Aphanomyces trifolii, Fusarium avenaceum and Rhizoctonia solani. Most breeding efforts have been devoted to resistance to P. clandestina, but the existence of different races has confounded selection. The most economically important virus diseases in subterranean clover pastures are Subterranean clover mottle virus and Bean yellow mosaic virus, while Subterranean clover stunt virus, Subterranean clover red leaf virus (local synonym for Soybean dwarf virus), Cucumber mosaic virus, Alfalfa mosaic virus, Clover yellow vein virus, Beet western yellows virus and Bean leaf roll virus also cause losses. Genotypic differences for resistance have been found to several of these fungal, oomycete and viral pathogens, highlighting the potential to develop cultivars with improved resistance. The most important pests of subterranean clover are redlegged earth mite (RLEM) (Halotydeus destructor), blue oat mite (Penthaleus major), blue-green aphid (Acyrthosiphon kondoi) and lucerne flea (Sminthurus viridis). New cultivars have been bred with increased RLEM cotyledon resistance, but limited selection has been conducted for resistance to other pests. Screening for disease and pest resistance has largely ceased, but recent molecular biology advances in subterranean clover provide a new platform for development of future cultivars with multiple resistances to important diseases and pests. However, this can only be realised if skills in pasture plant pathology, entomology, pre-breeding and plant breeding are maintained and adequately resourced. In particular, supporting phenotypic disease and pest resistance studies and understanding their significance is critical to enable molecular technology investments achieve practical outcomes and deliver subterranean clover cultivars with sufficient pathogen and pest resistance to ensure productive pastures across southern Australia.

Biology and trophic interactions of lucerne aphids

Abstract Lucerne or alfalfa Medicago sativa is the most important temperate forage legume worldwide. Only one or two varieties of lucerne were grown in the U.S.A. and Australia (the two leading exporters of lucerne) before the late 1950s and late 1970s, respectively. These dates coincided with the arrival of aphid species, which devastated lucerne stands and prompted the development of aphid resistant cultivars. Lucerne-feeding aphids, including bluegreen aphids Acyrthosiphon kondoi, pea aphids Acyrthosiphon pisum, spotted alfalfa aphids Therioaphis trifolii maculata and cowpea aphids Aphis craccivora, however, still present significant risks for the lucerne industry worldwide and account for 25% of global production losses. Moreover, increased production costs, negative environmental effects and emerging aphid resistance to insecticide applications have led to a narrowing of management options against lucerne aphids. Understanding lucerne aphid biology and trophic ecology will be needed to develop future management practices, including biological control. We review and synthesize research on the four lucerne aphid species, focussing on cultivar resistance and their interactions with other organisms, including predators, parasitoids, entomopathogens and bacterial symbionts. The effects of global climate change are considered, with a particular emphasis on the potential for compromised aphid resistance in lucerne cultivars under future climates. We conclude by identifying future research questions and perspectives for the sustainable management of lucerne aphids. These include the characterization of plant secondary metabolites associated with natural enemy recruitment, an understanding of the role of endosymbionts in cultivar resistance and a better comprehension of multi-trophic interactions of lucerne aphids, both with other herbivores and higher trophic groups.

Identification of distinct quantitative trait loci associated with defence against the closely related aphids Acyrthosiphon pisum and A. kondoi in Medicago truncatula

Aphids are a major family of plant insect pests. Medicago truncatula and Acyrthosiphon pisum (pea aphid, PA) are model species with a suite of resources available to help dissect the mechanism underlying plant–aphid interactions. A previous study focused on monogenic and relatively strong resistance in M. truncatula to PA and other aphid species. In this study a moderate resistance to PA was characterized in detail in the M. truncatula line A17 and compared with the highly susceptible line A20 and the more resistant line Jester. The results show that PA resistance in A17 involves both antibiosis and tolerance, and that resistance is phloem based. Quantitative trait locus (QTL) analysis using a recombinant inbred line (RIL) population (n=114) from a cross between A17 and A20 revealed that one locus, which co-segregated with AIN (Acyrthosiphon-induced necrosis) on chromosome 3, is responsible for the reduction of aphid biomass (indicator of antibiosis) for both PA and bluegreen aphid (BGA, A. kondoi), albeit to a lesser degree for PA than BGA. Interestingly, two independent loci on chromosomes 5 and 3 were identified for the plant biomass reduction (indicator of plant tolerance) by PA and BGA, respectively, demonstrating that the plant’s tolerance response to these two closely related aphid species is distinct. Together with previously identified major resistant (R) genes, the QTLs identified in this study are powerful tools to understand fully the spectrum of plant defence against sap-sucking insects and provide opportunities for breeders to generate effective and sustainable strategies for aphid control.

First Report of Alfalfa mosaic virus Infecting Basil (Ocimum basilicum) in California.

Basil (Ocimum basilicum L.) plants collected from three fields in Imperial County, CA in May, 2011 were found to be exhibiting yellowing, chlorotic sectors and spots on leaves, resulting in unmarketable plants. Dodder (Cuscuta spp.) was present in one of the fields, but was not visibly associated with symptomatic plants. Total nucleic acid was extracted from four symptomatic and three asymptomatic basil plants, as well as from the dodder plant with the RNeasy Plant Mini Kit (Qiagen, Valencia, CA). Nucleic acid extracts were tested by reverse transcription (RT)-PCR for the presence of Alfalfa mosaic virus (AMV) using primers designed to amplify a 350-nt region of the AMV coat protein gene (3). RT-PCR produced bands of the expected size in extracts from all symptomatic plants and the dodder sample. No amplification was obtained from symptomless plants. A 350-nt band amplified from one plant was gel-extracted, sequenced (TACGen, Richmond, CA), and confirmed to be AMV by comparison to sequences available in GenBank (Accession No. K02703). Although serological tests on an initial basil sample were negative for AMV by ELISA using antiserum produced against AMV by R. Larsen, USDA-ARS, Prosser, WA (unpublished), AMV was confirmed by ELISA and RT-PCR in symptomatic Nicotiana benthamiana, N. clevelandii, and Malva parviflora plants following mechanical transmission from basil source plants. The fields with AMV infections were located at opposite ends of the production region from one another, indicating widespread dispersal of AMV in the region. All AMV positive plants were adjacent to alfalfa. Two additional basil plantings in shade houses open to the outside environment did not have AMV symptomatic plants and were also confirmed negative by RT-PCR, but these plantings were at the extreme north end of Imperial Valley agriculture and well away from any alfalfa fields. At the time the basil plantations were sampled for AMV, no aphids were found in any plantations, but during the several weeks prior to finding the AMV-positive plants, cowpea aphid, Aphis craccivora Koch; pea aphid, Acyrthosiphon pisum Harris; blue alfalfa aphid, Acyrthosiphon kondoi Shinji; and spotted alfalfa aphid, Therioaphis maculata Buckton were colonizing Imperial Valley alfalfa fields, producing winged adults. AMV is transmitted by at least 14 aphid species (1), and most aphid populations increase during the late spring in this important desert agricultural region. The acquisition of AMV by dodder suggests the parasitic plant may serve as a vector of AMV within basil fields, although further study will be necessary for clarification. Significant acreage of basil is grown in the Imperial Valley. This acreage is surrounded by extensive and increasing alfalfa production totaling 55,442 ha (137,000 acres) in Imperial County and representing a 21% increase in acreage over 2009 for the same region (2). To our knowledge, this is the first report of basil infected by AMV in California. The proximity of basil production to such a large alfalfa production region warrants the need for enhanced efforts at aphid management in basil production to reduce vector populations and reduce transmission to basil crops.

Bean leafroll virus is widespread in subterranean clover (Trifolium subterraneum L.) seed crops and can be persistently transmitted by bluegreen aphid (Acyrthosiphon kondoi Shinji)

In the mid 2000s subterranean clover (Trifolium subterraneum) seed producers in South Australia reported symptoms of a red-leaf disease in fields with reduced seed yields. The red-leaf symptoms resembled those caused by several clover-infecting viruses. A set of molecular diagnostic tools were developed for the following viruses which are known to infect subterranean clover: Alfalfa mosaic virus; Bean leafroll virus (BLRV); Beet western yellows virus; Bean yellow mosaic virus; Cucumber mosaic virus; Pea seed-borne mosaic virus; Soybean dwarf virus and Subterranean clover stunt virus. Surveys of subterranean clover seed production fields in 2008 in the south-east of South Australia and western Victoria identified Bean leafroll virus, Alfalfa mosaic virus and Cucumber mosaic virus as present, with BLRV the most widespread. Surveys of pasture seed production fields and pasture evaluation trials in 2009 confirmed that BLRV was widespread. This result will allow seed producers to determine whether control measures directed against BLRV will overcome their seed losses. Bluegreen aphid (Acyrthosiphon kondoi) was implicated as a potential vector of BLRV because it was observed to be colonising lucerne plants adjacent to subterranean clover seed production paddocks with BLRV, and in a glasshouse trial it transmitted BLRV from an infected lucerne plant to subterranean clover in a persistent manner.