Insect Science

Candidatus Liberibacter asiaticus (CLas) is a phloem-limited bacterium that is associated with the Huanglongbing (HLB) disease of citrus and transmitted by the psyllid, Diaphorina citri. There are no curative methods to control HLB and the prevention of new infections is essential for HLB management. Therefore, the objective of our study was to determine the effects of systemic insecticides, such as the neonicotinoids imidacloprid, thiamethoxam, and a mixture of thiamethoxam and chlorantraniliprole (diamide) on the probing behavior of CLas-infected D. citri and their effect on CLas transmission. The electrical penetration graph (EPG-DC) technique was used to monitor the stylet penetration activities of CLas-infected D. citri on sweet orange [Citrus sinensis (L.) Osbeck] ‘Valencia’ treated with systemic insecticides. Systemic insecticides disrupted the probing behavior of CLas-infected D. citri, in a way that affected CLas transmission efficiency, particularly by negatively affecting the stylet activities related to the phloem phase. All insecticides reduced (by 57–73%) the proportion of psyllids that exhibited sustainable phloem ingestion (waveform E2 > 10 min), with significant differences observed on plants treated with thiamethoxam and thiamethoxam + chlorantraniliprole. The transmission rate of CLas with high inoculum pressure (five CLas-infected D. citri per plant and a seven-day inoculation access period) to untreated control plants was 93%. In contrast, CLas transmission was reduced to 38.8% when test plants were protected by systemic insecticides. Our results indicated that all insecticides tested presented a potential to reduce CLas inoculation by an average of 59%; therefore, these insecticides can be used to reduce the spread of HLB.

The tomato potato psyllid (TPP), Bactericera cockerelli, is a psyllid native to North America that has recently invaded New Zealand and Australia. The potential for economic losses accompanying invasions of TPP and its associated bacterial plant pathogen Candidatus Liberibacter solanacearum (CLso), has caused much concern. Here, we employed ecological niche models to predict environments suitable for TPP/CLso on a global scale and then evaluated the extent to which global potato cultivation is at risk. In addition, at a finer scale the risk to the Australian potato acreage was evaluated. A total of 86 MaxEnt models were built using various combinations of settings and climatic predictors, and the best model based on model evaluation metrics was selected. Climatically suitable habitats were identified in Eurasia, Africa, South America, and Australasia. Intersecting the predicted suitability map with land use data showed that 79.06% of the global potato cultivation acreage, 96.14% of the potato production acreage in South America and Eurasia, and all the Australian potato cropping areas are at risk. The information generated by this study increases knowledge of the ecology of TPP/CLso and can be used by government agencies to make decisions about preventing the spread of TPP and CLso across the globe.

AbstractHuanglongbing, a highly destructive disease of citrus species, is associated with a fastidious, gram-negative, phloem-limited bacteria (Candidatus Liberibacter spp.). In Florida, the causative agent of Huanglongbing (HLB) is C. Liberibacter asiaticus (CLas) and it is transmitted by the insect vector, Asian citrus psyllid (Diaphorina citri Kuwayama). Previous investigations have revealed systemic infection of CLas with an erratic and uneven distribution of pathogen in tree phloem. However, previous investigations did not consider the potential impact of plant vegetative growth on presence/absence of CLas in planta. Our objectives were to determine: 1) the effect of vegetative growth of Citrus sinensis (L.) Osbeck cv Valencia on detection of CLas in mature leaves, and 2) the impact of CLas inoculation frequency on progression of CLas titer in citrus leaves through the first year of infection. Temporal dynamics of CLas detection were associated with vegetative flush growth. Surprisingly, there was no difference in CLas titer detected between plants exposed to infected vectors for a one-time 7 d inoculation access period, as compared with plants exposed to continuously breeding CLas-infected insects over the course of an entire year of plant infection. Our results suggest that the CLas bacterium is transported through phloem during annual movement of carbon compounds needed for vegetative plant growth, including transportation from roots to mature leaves. These results highlight the importance of vegetative growth on temporal dynamics of CLas in citrus, and suggest a critical role of the sink-source interaction on presence/absence of CLas in leaves.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an important transmission vector of the citrus greening disease Candidatus Liberibacter asiaticus (CLas). The D. citri midgut exhibits an important tissue barrier against CLas infection. However, the molecular mechanism of the midgut response to CLas infection has not been comprehensively elucidated. In this study, we identified 778 differentially expressed genes (DEGs) in the midgut upon CLas infection, by comparative transcriptome analyses, including 499 upregulated DEGs and 279 downregulated DEGs. Functional annotation analysis showed that these DEGs were associated with ubiquitination, the immune response, the ribosome, endocytosis, the cytoskeleton and insecticide resistance. KEGG enrichment analysis revealed that most of the DEGs were primarily involved in endocytosis and the ribosome. A total of fourteen DEG functions were further validated by reverse transcription quantitative PCR (RT-qPCR). This study will contribute to our understanding of the molecular interaction between CLas and D. citri.

Abstract The potato/tomato psyllid Bactericera cockerelli (Šulc) transmits ‘Candidatus Liberibacter solanacearum’ (Lso) (also known as ‘Candidatus Liberibacter psyllaurous’), the bacterium associated with zebra chip disease (ZC) in potato. When disease incidence is high, ZC causes large economic losses through reductions in potato yield and tuber quality. No commercial potato variety has been found totally resistant to the pathogen. We evaluated host acceptance behaviors using no-choice assays on three breeding clones derived from Solanum chacoense Bitter with putative tolerance to Lso and/or ZC as part of an effort to determine whether the disease tolerance observed in those breeding clones was related to effects on psyllid settling behavior. We also counted the number of eggs laid and nymphs hatched on the different genotypes to observe any differences in reproduction. The potato variety ‘Russet Burbank’ was used as a susceptible control. Probing frequency and female walking duration were greater on Russet Burbank than the other genotypes. Oviposition did not differ among genotypes. However, female psyllids on two of the Lso-tolerant genotypes displayed reduced fertility 18–24 d after confinement with a male, relative to females on Russet Burbank. These results suggest that although the germplasms display minor abiotic activity on psyllid fertility, tolerance to Lso may be more strongly linked with plant tolerance to the pathogen rather than effects on host acceptance behaviors.

Abstract“Candidatus Liberibacter solanacearum” (Lso) are phloem‐restricted and unculturable Gram‐negative bacteria. Presently five haplotypes have been identified worldwide; but only haplotypes A and B are associated with the vector Bactericera cockerelli (Šulc.) in the Americas. Previous studies showed that Lso‐infection reduces B. cockerelli reproductive output and that Lso haplotype B is more pathogenic than Lso haplotype A. To understand the interaction of Lso haplotype B and B. cockerelli, the fitness of Lso‐free and Lso B‐infected insects, and the expression of vitellogenin (BcVg1‐like), a gene involved directly in the insect reproduction were analyzed. Statistical differences in the number of eggs oviposited, and the total number of progeny nymphs and adults were found among crosses of insects with or without Lso. Significant differences in sex proportions were found between Lso B‐infected and Lso‐free crosses: a higher proportion of F1 adult females were obtained from Lso B‐infected mothers. A significant reduction of BcVg1‐like was observed in crosses performed with Lso B‐infected females compared to the Lso‐free insects. In female cohorts of different age, a significant reduction of BcVg1‐like expression was measured in 7‐d‐old Lso B‐infected females (virgin and mated) compared with 7‐d‐old Lso‐free females (virgin and mated), respectively. The reduction of BcVg1‐like transcript was associated with a lower number of developing oocytes observed in female's reproductive systems. Overall, this study represents the first step to understand the interaction of Lso B with B. cockerelli, highlighting the effect of Lso B infection on egg production, BcVg1‐like expression, and oocyte development.