Litao Guo

Genome sequencing of the sweetpotato whitefly Bemisia tabaci MED/Q

Abstract The sweetpotato whitefly Bemisia tabaci is a highly destructive agricultural and ornamental crop pest. It damages host plants through both phloem feeding and vectoring plant pathogens. Introductions of B. tabaci are difficult to quarantine and eradicate because of its high reproductive rates, broad host plant range, and insecticide resistance. A total of 791 Gb of raw DNA sequence from whole genome shotgun sequencing, and 13 BAC pooling libraries were generated by Illumina sequencing using different combinations of mate-pair and pair-end libraries. Assembly gave a final genome with a scaffold N50 of 437 kb, and a total length of 658 Mb. Annotation of repetitive elements and coding regions resulted in 265.0 Mb TEs (40.3%) and 20 786 protein-coding genes with putative gene family expansions, respectively. Phylogenetic analysis based on orthologs across 14 arthropod taxa suggested that MED/Q is clustered into a hemipteran clade containing A. pisum and is a sister lineage to a clade containing both R. prolixus and N. lugens. Genome completeness, as estimated using the CEGMA and Benchmarking Universal Single-Copy Orthologs pipelines, reached 96% and 79%. These MED/Q genomic resources lay a foundation for future ‘pan-genomic’ comparisons of invasive vs. noninvasive, invasive vs. invasive, and native vs. exotic Bemisia, which, in return, will open up new avenues of investigation into whitefly biology, evolution, and management.

Differential effects of an exotic plant virus on its two closely related vectors

Concurrent spread of Tomato yellow leaf curl virus (TYLCV) with invasion of Bemisia tabaci Q rather than B in China suggests a more mutualistic relationship between TYLCV and Q than B. To assess this hypothesis, we quantified the impacts of TYLCV on the performance and competitiveness of B and Q in the laboratory. The results showed that relative to their non-infected counterparts feeding on cotton (a non-host for TYLCV), infected B exhibited significant reductions in life-history traits, whereas infected Q only showed marginal reductions. While Q performed better on TYLCV-infected tomato plants than on uninfected ones, the reverse was observed in B. Q displacement by B took one more generation on TYLCV-infected tomato plants than on healthy ones. These results demonstrate that TYLCV was indirectly mutualistic to Q but directly and indirectly parasitic to B.

Sensitivity of Bemisia tabaci (Hemiptera: Aleyrodidae) to several new insecticides in China: effects of insecticide type and whitefly species, strain, and stage.

ABSTRACT. Whitefly biotypes B and Q are the two most damaging members of the Bemisia tabaci (Hemiptera: Aleyrodidae) species complex. Control of B. tabaci (and especially of Q) has been impaired by resistance to commonly used insecticides. To find new insecticides for B. tabaci management in China, we investigated the sensitivity of eggs, larvae, and adults of laboratory strains of B and Q (named Lab-B and Lab-Q) and field strains of Q to several insecticides. For eggs, larvae, and adults of B. tabaci and for six insecticides (cyantraniliprole, chlorantraniliprole, pyriproxyfen, buprofezin, acetamiprid, and thiamethoxam), LC50 values were higher for Lab-Q than for Lab-B; avermectin LC50 values, however, were low for adults of both Lab-Q and Lab-B. Based on the laboratory results, insecticides were selected to test against eggs, larvae, and adults of four field strains of B. tabaci Q. Although the field strains differed in their sensitivity to the insecticides, the eggs and larvae of all strains were highly sensitive to cyantraniliprole, and the adults of all strains were highly sensitive to avermectin. The eggs, larvae, and adults of B. tabaci Q were generally more resistant than those of B. tabaci B to the tested insecticides. B. tabaci Q eggs and larvae were sensitive to cyantraniliprole and pyriproxyfen, whereas B. tabaci Q adults were sensitive to avermectin. Field trials should be conducted with cyantraniliprole, pyriproxyfen, and avermectin for control of B. tabaci Q and B in China.

Glutathione S-transferases are involved in thiamethoxam resistance in the field whitefly Bemisia tabaci Q (Hemiptera: Aleyrodidae)

The whitefly, Bemisia tabaci, has developed a high level of resistance to thiamethoxam, a second generation neonicotinoid insecticide that has been widely used to control this pest. In this study, we assessed the level of cross-resistance, the activities of detoxifying enzymes, and the expression profiles of 23 glutathione S-transferase (GST) genes in a thiamethoxam-resistant ant and -susceptible strain of Bemisia tabaci Q. The thiamethoxam-resistant strain showed a moderate level of cross-resistance to another nicotinoid insecticide imidacloprid, a low level of cross-resistance to acetamiprid and nitenpyram, and no significant cross-resistance to abamectin and bifenthrin. Among detoxifying enzymes, only GSTs had significantly higher activity in the resistant strain than in the susceptible strain. Seven of 23 GST genes were over-expressed in the resistant strain relative to the susceptible strain. Using the technology of RNA interference to knockdown a GST gene (GST14), the results showed that silencing GST14 increased the mortality of whiteflies to thiamethoxam in Bemisia tabaci.

Detoxification enzymes of Bemisia tabaci B and Q: biochemical characteristics and gene expression profiles.

BACKGROUND The sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is one of the most invasive and destructive pests of field crops worldwide. The sibling species B and Q are the two most damaging members of the B. tabaci species complex. That Q is more resistant than B to many insecticides has been well documented. Over the last decade, Q has gradually displaced B and has become the dominant form of B. tabaci in field agricultural systems in most parts of China. To help understand the differences in insecticide resistance, the activities and gene expression profiles of detoxification enzymes in B. tabaci B and Q were investigated. RESULTS The activity of P450 towards 7-ethoxycoumarin was significantly higher (1.46-fold higher) in Q than in B. The expression of 43 of 65 P450 genes was higher (>1-fold) in Q than in B, and expression for eight P450 genes was more than 50-fold greater in Q than in B. The increased expression of selected P450 genes in Q relative to B was confirmed with two other B strains and two other Q strains. On the other hand, carboxylesterase (CarE) activity was significantly lower (0.71-fold lower) in Q than in B; the Km value of CarE was significantly lower in B than in Q, but the opposite was true for the Vmax value of CarE. Glutathione S-transferase activity and values of Km and Vmax did not differ between B and Q. CONCLUSION Enhanced metabolic detoxification of insecticides by P450s may be an important reason why B. tabaci Q is more resistant than B. tabaci B to insecticides.

Transcriptomic dissection of sexual differences in Bemisia tabaci , an invasive agricultural pest worldwide

Sex difference involving chromosomes and gene expression has been extensively documented. In this study, the gender difference in the sweetpotato whitefly Bemisia tabaci was investigated using Illumina-based transcriptomic analysis. Gender-based RNAseq data produced 27 Gb reads, and subsequent de novo assembly generated 93,948 transcripts with a N50 of 1,853 bp. A total of 1,351 differentially expressed genes were identified between male and female B. tabaci, and majority of them were female-biased. Pathway and GO enrichment experiments exhibited a gender-specific expression, including enriched translation in females, and enhanced structural constituent of cuticle in male whiteflies. In addition, a putative transformer2 gene (tra2) was cloned, and the structural feature and expression profile of tra2 were investigated. Sexually dimorphic transcriptome is an uncharted territory for the agricultural insect pests. Molecular understanding of sex determination in B. tabaci, an emerging invasive insect pest worldwide, will provide potential molecular target(s) for genetic pest control alternatives.

Differing effects of cabbage and pepper on B and Q putative species of Bemisia tabaci

B and Q putative species of Bemisia tabaci are two of the most invasive pests in the world. In China, Q is displacing B, but the underlying mechanism has been poorly explored. The difference in tolerance to host plants between B and Q may shed some light on the displacement. In the present study, two types of resistance (antixenosis or antibiosis) of cabbage and pepper against B and Q putative species were characterized based on demographic parameters (host selection, fecundity, etc.) and feeding behavior [as determined by electrical penetration graph (EPG)]. Female longevity, fecundity, and nymph survival were substantially higher on cabbage than on pepper for B, but were similar on the two hosts for Q. B nymphs suffered high mortality on pepper, suggesting a high level of antibiosis against B in pepper. Both B and Q preferred to settle and oviposit on cabbage rather than on pepper, indicating a high level of antixenosis against B and Q in pepper. With regard to feeding behavior, EPG data indicated that both B and Q whiteflies fed more efficiently on cabbage than on pepper. Overall, both B and Q preferred cabbage to pepper, and both performed better on cabbage than on pepper, but the negative effects of pepper were greater on B than on Q. Our results demonstrate that Q is more tolerant of a low suitability host than B, which may partially explain why Q has displaced B in China.

Transcriptome analysis of host-associated differentiation in Bemisia tabaci (Hemiptera: Aleyrodidae)

Host-associated differentiation is one of the driving forces behind the diversification of phytophagous insects. In this study, host induced transcriptomic differences were investigated in the sweetpotato whitefly Bemisia tabaci, an invasive agricultural pest worldwide. Comparative transcriptomic analyses using coding sequence (CDS), 5′ and 3′ untranslated regions (UTR) showed that sequence divergences between the original host plant, cabbage, and the derived hosts, including cotton, cucumber and tomato, were 0.11–0.14%, 0.19–0.26%, and 0.15–0.21%, respectively. In comparison to the derived hosts, 418 female and 303 male transcripts, respectively, were up-regulated in the original cabbage strain. Among them, 17 transcripts were consistently up-regulated in both female and male whiteflies originated from the cabbage host. Specifically, two ESTs annotated as Cathepsin B or Cathepsin B-like genes were significantly up-regulated in the original cabbage strain, representing a transcriptomic response to the dietary challenges imposed by the host shifting. Results from our transcriptome analysis, in conjunction with previous reports documenting the minor changes in their reproductive capacity, insecticide susceptibility, symbiotic composition and feeding behavior, suggest that the impact of host-associated differentiation in whiteflies is limited. Furthermore, it is unlikely the major factor contributing to their rapid range expansion/invasiveness.

RNA interference-mediated knockdown of the hydroxyacid-oxoacid transhydrogenase gene decreases thiamethoxam resistance in adults of the whitefly Bemisia tabaci

Bemisia tabaci has developed a high level of resistance to thiamethoxam, a second generation neonicotinoid insecticide that has been widely used to control this pest. In this study, we investigated whether hydroxyacid-oxoacid transhydrogenase (HOT) is involved in resistance to the neonicotinoid insecticide thiamethoxam in the whitefly. We cloned the full-length gene that encodes HOT in B. tabaci. Its cDNA contains a 1428-bp open reading frame encoding 475 amino acid residues. Then we evaluated the mRNA expression level of HOT in different developmental stages, and found HOT expression was significantly greater in thiamethoxam resistance adults than in thiamethoxam susceptible adults. Subsequently, seven field populations of B. tabaci adults were sampled, the expression of mRNA level of HOT significant positive correlated with thiamethoxam resistance level. At last, we used a modified gene silencing system to knock-down HOT expression in B. tabaci adults. The results showed that the HOT mRNA levels decreased by 57% and thiamethoxam resistance decreased significantly after 2 days of feeding on a diet containing HOT dsRNA. The results indicated that down-regulation of HOT expression decreases thiamethoxam resistance in B. tabaci adults.

Molecular cloning of the sex-related gene PSI in Bemisia tabaci and its alternative splicing properties

The P-element somatic inhibitor (PSI) is gene known to regulate the transcription of doublesex (dsx) when transformer (tra) is absent in Bombyx mori. In this study, we identified and characterized a PSI homolog in Bemisia tabaci (BtPSI). BtPSI cDNA had a total length of 5700 bp and contained a predicted open reading frame (ORF) of 2208 nucleotides encoding for 735 amino acids. Multiple sequence alignments of the common regions of PSI proteins from B. tabaci and five other insect species revealed a high degree of sequence conservation. BtPSI is expressed in all stages of B. tabaci development, and expression did not significantly differ between female and male adult. A total of 92 BtPSI isoforms (78 in female and 22 in male) were identified, and a marker indicating the female-specific form was found. These results increase the understanding of genes that may determine sex in B. tabaci and provide a foundation for research on the sex determination mechanism in this insect.