Chromosome-level genome assembly of the Asian citrus psyllid,Diaphorina citri

Diaphorina citri is a global citrus pest.As a vector insect,it can transmit the causative agents of citrus huanglongbing,causing irreversible losses to the citrus industry.The acquisition of genomic information can provide a molecular genetic basis for effective control of D.citri.Here,the DNBSEQ™,Oxford Nanopore Technologies,and Hi-C technologies are applied to generate a high-quality chromosome-level genome of D.citri.The genome size of D.citri was 523.78 Mb with a scaffold N50 of 47.05 Mb distributed on 13 chromosomes.A total of 250.64 Mb(47.85%)repeat sequences and 24048 protein-coding genes were predicted.Genome resequencing of female and male individuals indicated that the sex chromosome system of D.citri is XO.Phylogenetic analysis demonstrated that D.citri and Pachypsylla venusta,which separated from their most recent common ancestor about 336.62 million years ago,were the most closely related.Additionally,we identified genes potentially involved in detoxification metabolism,pathogen transmission,and honeydew secretion for further investigation.The high-quality genome provides an important reference for developing effective management strategies of D.citri.

Transcriptomic analysis reveals the defense mechanisms of citrus infested with Diaphorina citri

Huanglongbing(HLB) is a devastating disease that has led to an acute crisis for growers of citrus, one of the world's most important fruit crops. The phloem-feeding Asian citrus psyllid(ACP), Diaphorina citri, is the main pest at the new shoot stage and is the only natural vector of HLB pathogenic bacteria. Little is known about how plants perceive and defend themselves from this destructive pest. Here, we characterized changes in the expression of various genes in citrus plants that were continuously infested by D. citri for different durations(12, 24, and 48 h). A total of 5 219 differentially expressed genes(DEGs) and 643 common DEGs were identified across all time points. Several pathways related to defense were activated, such as peroxisome, alpha-linolenic acid metabolism, and phenylpropanoid and terpenoid biosynthesis, and some pathways related to growth and signal transduction were suppressed in response to D. citri infestation. The expression of genes including kinases(CML44, CIPK6, and XTH6), phytohormones(SAMT, LOX6, and NPR3), transcription factors(bHLH162, WRKY70, and WRKY40), and secondary metabolite synthesis-related genes(PAL, 4CL2, UGT74B1 and CYP82G1) was significantly altered in response to D. citri infestation. The findings of this study greatly enhance our understanding of the mechanisms underlying the defense response of citrus plants to D. citri infestation at the molecular level. Functional characterization of the candidate defense-related genes identified in this study will aid the molecular breeding of insect-resistant citrus varieties.

Repellent effect of guava leaf volatiles on settlement of adults of citrus psylla, Diaphorina citri Kuwayama, on citrus

The Asiatic citrus psyllid （Diaphorina cirri Kuwayama [Hemiptera： Sternorrhyncha： Psyllidae] is a vector ofhuanglongbing （citrus greening）, a devastating disease of citrus caused by phloem-limited bacteria. Growing guava （Psidium guajava） as an intercrop appears to be a successful means of reducing psyllid numbers within citrus orchards; however, the mechanism by which such a reduction is achieved is unknown. To determine the repellent effect of guava leaf and factors attributed to this activity, responses of adult psyllids to guava leaf and its odor were evaluated in cage tests and Y-tube olfactometer test. The results showed that guava leaf possessed a repellent effect against the adult citrus psyllids. Fewer psyllids were found on citrus leaves in the presence of guava foliage than in its absence. Young and old guava leaf showed equal repellent activity. By covering the guava shoots with net cloth, it was revealed that the repellent effect of guava leaf against adult psyllids on citrus was attributed to the volatile compounds, rather than physical factors. The olfactometer response of adult psyllids to guava leaf odor was dosage-dependent. Between guava odor and control, only 35.00%, 25.00% and 16.25% of the psyllids moved toward guava odor when presented with 5.0, 10.0 and 15.0 g of guava shoots, respectively. The olfactometer experiments also showed that both male and female psyllids responded similarly to the guava leaf odor.

Botanicals, selective insecticides, and predators to control Diaphorina citri （Hemiptera： Liviidae） in citrus orchards

The Asian citrus psyllid （ACP） Diaphorina citri Kuwayama vectors pathogens that cause huanglongbing （HLB） or citrus greening devastating and economically im- portant disease present in most citrus growing regions. Young citrus shoots are required for psyllid reproduction and development. During winter citrus trees produce little or no new growth. Overwintering adults reproduce in spring on newly emerging shoots also attractive to other pests and beneficial insects. Botanicals and relatively selective insecti- cides could help to conserve beneficial insects and reduce pest resistance to insecticides. Sprays of Azadirachtin （Neem）, Tropane （Datura）, Spirotetramat, Spinetoram, and broad- spectrum Imidacloprid were evaluated to control ACP in spring and summer on 10-year-old ＂Kinow＂ Citrus reticulata Blanco trees producing new growth. Psyllid populations were high averaging 5-9 nymphs or adults per sample before treatment application. Nymphs or adults were significantly reduced to 0.5-1.5 per sample in all treatments for 3 weeks, aver- age 61%-83% reduction. No significant reduction in ladybeetles Adalia bipunctata, Ane- glei scardoni, Cheilomenes sexmaculata, and Coccinella septempunctata was observed. Syrphids, spiders and green lacewings were reduced in treated trees except with Tropane. Studies are warranted to assess impact of these predators on ACP and interaction with insecticides. Observed reduction in ACP populations may not be enough considering its reproductive potential and role in the spread of HLB. Follow-up sprays may be required to achieve additional suppression using rotations of different insecticides.

Selection of Bacillus thuringiensis strains in citrus and their pathogenicity to Diaphorina citri (Hemiptera: Liviidae) nymphs

Bacillus thuringiensis(Bt)toxins are effective in controlling insect pests either through the spraying of products or when expressed in transgenic crops.The discovery of endophytic Bt strains opened new perspectives for studies aimed at the control of sap‐sucking insects,such as the Asian citrus psyllid Diaphorina citri Kuwayama(Hemiptera:Liviidae),a vector of“Candidatus Liberibacter spp.,”associated with citrus huanglongbing(HLB).In this study,translocation of endophytic Bt strains in citrus seedlings inoculated with Bt suspension delivered by soil‐drench,and their systemic pathogenicity to D.citri nymphs were investigated.The pathogenicity of three wild‐type Bt strains against D.citri third‐instar nymphs was demonstrated.Among the 10 recombinant strains tested(each of them harboring a single cry or cyt gene),3 can be highlighted,causing 42%–77%and 66%–90%nymphal mortality at 2 and 5 d after inoculation,respectively.The isolation of Bt cells from young citrus shoots and dead nymphs,and PCR performed with specific primers,confirmed the involvement of the Bt strains in the psyllid mortality.This is the first report showing the translocation of Bt strains from citrus seedling roots to shoots and their potential to control D.citri nymphs that fed on these soil‐drench inoculated seedlings.The Bt strains that caused the highest mortality rates have the potential to be used as bioinsecticides to control D.citri and the identified genes can be used for the production of transgenic Bt citrus.

Huanglongbing:An overview of a complex pathosystem ravaging the world's citrus

Citrus huanglongbing（HLB） has become a major disease and limiting factor of production in citrus areas that have become infected. The destruction to the affected citrus industries has resulted in a tremendous increase to support research that in return has resulted in significant information on both applied and basic knowledge concerning this important disease to the global citrus industry. Recent research indicates the relationship between citrus and the causal agent of HLB is shaped by multiple elements, in which host defense responses may also play an important role. This review is intended to provide an overview of the importance of HLB to a wider audience of plant biologists. Recent advances on host-pathogen interactions, population genetics and vectoring of the causal agent are discussed.