Discovery of Brassica Yellows Virus and Porcine Reproductive and Respiratory Syndrome Virus in Diaphorina citri and Changes in Virome Due to Infection with 'Ca. L. asiaticus'.

Detection of new viruses or new virus hosts is essential for the protection of economically important agroecosystems and human health. Increasingly, metatranscriptomic data are being used to facilitate this process. Such data were obtained from adult Asian citrus psyllids (ACP) (Diaphorina citri Kuwayama) that fed solely on mandarin (Citrus ×aurantium L.) plants grafted with buds infected with 'Candidatus Liberibacter asiaticus' (CLas), a phloem-limited bacterium associated with the severe Asian variant of huanglongbing (HLB), the most destructive disease of citrus. Brassica yellows virus (BrYV), the causative agent of yellowing or leafroll symptoms in brassicaceous plants, and its associated RNA (named as BrYVaRNA) were detected in ACP. In addition, the porcine reproductive and respiratory syndrome virus (PRRSV), which affects pigs and is economically important to pig production, was also found in ACP. These viruses were not detected in insects feeding on plants grafted with CLas-free buds. Changes in the concentrations of insect-specific viruses within the psyllid were caused by coinfection with CLas. IMPORTANCE The cross transmission of pathogenic viruses between different farming systems or plant communities is a major threat to plants and animals and, potentially, human health. The use of metagenomics is an effective approach to discover viruses and vectors. Here, we collected buds from the CLas-infected and CLas-free mandarin (Citrus ×aurantium L. [Rutaceae: Aurantioideae: Aurantieae]) trees from a commercial orchard and grafted them onto CLas-free mandarin plants under laboratory conditions. Through metatranscriptome sequencing, we first identified the Asian citrus psyllids feeding on plants grafted with CLas-infected buds carried the plant pathogen, brassica yellows virus and its associated RNA, and the swine pathogen, porcine reproductive and respiratory syndrome virus. These discoveries indicate that both viruses can be transmitted by grafting and acquired by ACP from CLas+ mandarin seedlings.

Integrated Pest Management Strategies for Asian Citrus Psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae) and Huanglongbing in Citrus for Sarawak, East Malaysia, Borneo.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, transmits 'Candidatus Liberibacter asiaticus' (CLas), a phloem-limited bacterium associated with the severe Asian form of huanglongbing (HLB), and the most destructive disease of citrus. The pathogen and the psyllid, both of South Asian origin, are now widespread in citrus regions of Asia and the Americas. There is no cure for the disease. Application of synthetic pesticides, in some instances more frequently than fortnightly, to minimise incidence of ACP in citrus orchards, has not prevented inevitable impacts of the disease in regions of Asia where CLas is present. Despite the inevitable spread of the disease, significant progress has been made in Sarawak since the mid-1990s towards effectively implementing integrated pest management (IPM) programs for stemming the impact of the disease and detrimental consequences of over-reliance on synthetic pesticides. Growers are encouraged to plant pathogen-free trees, remove diseased trees, monitor incidence of the psyllid, and to use pesticides judiciously to reduce their detrimental impacts on natural enemies. Knowledge has been enhanced through research on seasonal incidence of the psyllid, use of mineral oils, development of protocols and iodine−starch test kits for detecting infected trees, PCR for confirming the presence of CLas in symptomatic leaves, methods for monitoring incidence the psyllid, and training extension staff and growers. However, major impediments to increasing the average longevity of trees beyond <5 years in poorly managed orchards, based on marcotting (air layering), and >12 years in well-managed orchards, based on pathogen-free trees, still need to be addressed. These include grower knowledge, marcotting, aggressive marketing of synthetic pesticides, high prices of mineral oils, spray application procedures, and better reliance on natural enemies of the psyllid.

Genome Sequence Resource of 'Candidatus Liberibacter asiaticus' from Diaphorina citri Kuwayama (Hemiptera: Liviidae) from La Réunion.

'Candidatus Liberibacter asiaticus' is an insect-transmitted, phloem-restricted α-proteobacterium associated with huanglongbing. Here, we provide the whole genome sequence of 'Ca. L. asiaticus' strain ReuSP1 from its insect vector Diaphorina citri (Hemiptera: Liviidae) collected in La Réunion. The genome is composed of 1,230,064 bp and has a 36.5% G+C content. This study reports the first 'Ca. L. asiaticus' genome sequence from La Réunion, which will add to 'Ca. L. asiaticus' genome resources and help elucidate our understanding of the introduction pathway into La Réunion.

Draft Genome Sequence of a Novel "Candidatus Liberibacter" Species Detected in a Zanthoxylum Species from Bhutan.

The draft genome sequence of a novel "Candidatus Liberibacter" species detected in an unidentified species of Zanthoxylum (Rutaceae) collected in Bhutan is reported. The total length is 1,408,989 bp with 1,169 coding sequences in 96 contigs, a GC content of 37.3%, and 76 to 77% average nucleotide identity with several other "Ca Liberibacter" species.

Characterization of the bacterial communities of psyllids associated with Rutaceae in Bhutan by high throughput sequencing.

BACKGROUND: Several plant-pathogenic bacteria are transmitted by insect vector species that often also act as hosts. In this interface, these bacteria encounter plant endophytic, insect endosymbiotic and other microbes. Here, we used high throughput sequencing to examine the bacterial communities of five different psyllids associated with citrus and related plants of Rutaceae in Bhutan: Diaphorina citri, Diaphorina communis, Cornopsylla rotundiconis, Cacopsylla heterogena and an unidentified Cacopsylla sp. RESULTS: The microbiomes of the psyllids largely comprised their obligate P-endosymbiont 'Candidatus Carsonella ruddii', and one or two S-endosymbionts that are fixed and specific to each lineage. In addition, all contained Wolbachia strains; the Bhutanese accessions of D. citri were dominated by a Wolbachia strain first found in American isolates of D. citri, while D. communis accessions were dominated by the Wolbachia strain, wDi, first detected in D. citri from China. The S-endosymbionts from the five psyllids grouped with those from other psyllid taxa; all D. citri and D. communis individuals contained sequences matching 'Candidatus Profftella armatura' that has previously only been reported from other Diaphorina species, and the remaining psyllid species contained OTUs related to unclassified Enterobacteriaceae. The plant pathogenic 'Candidatus Liberibacter asiaticus' was found in D. citri but not in D. communis. Furthermore, an unidentified 'Candidatus Liberibacter sp.' occurred at low abundance in both Co. rotundiconis and the unidentified Cacopsylla sp. sampled from Zanthoxylum sp.; the status of this new liberibacter as a plant pathogen and its potential plant hosts are currently unknown. The bacterial communities of Co. rotundiconis also contained a range of OTUs with similarities to bacteria previously found in samples taken from various environmental sources. CONCLUSIONS: The bacterial microbiota detected in these Bhutanese psyllids support the trends that have been seen in previous studies: psyllids have microbiomes largely comprising their obligate P-endosymbiont and one or two S-endosymbionts. In addition, the association with plant pathogens has been demonstrated, with the detection of liberibacters in a known host, D. citri, and identification of a putative new species of liberibacter in Co. rotundiconis and Cacopsylla sp.