Skip to main content
Download PDF

Comparative proteomic analysis provides new insight into differential transmission of two begomoviruses by a whitefly

Virology Journal volume 16, Article number: 32 (2019) Cite this article

Abstract

Background

Viruses in the genus Begomovirus (Family Geminiviridae) include many important economic plant viruses transmitted by whiteflies of the Bemisia tabaci species complex. In general, different begomoviruses may be acquired and transmitted by the same whitefly species with different efficiencies. For example, the species Mediterranean (MED) in this whitefly species complex transmits tomato yellow leaf curl virus (TYLCV) at a higher efficiency than papaya leaf curl China virus (PaLCuCNV). However, the proteomic responses of whitefly to the infection of different begomoviruses remain largely unknown.

Methods

We used iTRAQ-based proteomics coupled with RT-qPCR to investigate and compare responses of the MED whitefly to the infection of TYLCV and PaLCuCNV.

Results

Totally, 259, 395 and 74 differently expressed proteins (DEPs) were identified in the comparisons of TYLCV-infected vs. un-infected, PaLCuCNV-infected vs. un-infected, and TYLCV-infected vs. PaLCuCNV-infected whiteflies, respectively. These proteins appear associated with catabolic process, metabolic process, transport, defense response, cell cycle, and receptor. The comparisons of TYLCV-infected vs. un-infected and PaLCuCNV-infected vs. un-infected shared some similar DEPs, indicating possible involvement of laminin subunit alpha, dystroglycan, integrin alpha-PS2 and cuticle proteins in viral transport as well as the role of putative defense proteins 3 and PITH in anti-viral response. However, 20S proteasome subunits associated with regulation of virus degradation and accumulation were up-regulated in PaLCuCNV-infected but not in TYLCV-infected whiteflies, which may be related to the constraints of PaLCuCNV accumulation in MED.

Conclusions

These findings provide valuable clues for unravelling the roles of some whitefly proteins in begomovirus transmission.

Background

Plant diseases caused by begomoviruses have been major constraints to the production of many economic crops such as tomato and cotton [1]. Begomoviruses are a group of single-stranded circular DNA viruses with twinned particles, and their genetic structure is bipartite or monopartite [2]. So far, more than 388 species have been described in the genus Begomovirus (Family Geminiviridae) [3], which in general are transmitted by whiteflies in the Bemisia tabaci species complex in a persistent, circulative manner [4]. For begomoviruses, once orally acquired by whitefly, they follow the path of head-midgut-haemolymph-primary salivary gland inside the vector [5,6,7]. The coat protein is the only known viral structural protein in determining begomovirus transmission characteristics [8]. In the whitefly, two organs including midgut and primary salivary gland have been identified as barriers in the circulative journey of begomovirus in the vector body [9,10,11]. A few proteins have been investigated for their roles in begomovirus transmission. Two whitefly proteins, cyclophilin B and a midgut protein, seem to have roles in assisting begomovirus transmission [12, 13]; while another two whitefly proteins, heat shock protein 70 and Knottin-1, seem to negatively affect begomovirus transmission by whitefly [14, 15]. In addition, a peptidoglycan recognition protein and an antimicrobial peptide have been reported to be involved in whitefly-begomovirus interaction [16, 17].

Tomato yellow leaf curl virus (TYLCV) is a monopartite begomovirus without satellite DNA and is one of the most economically important begomoviruses all over the world [1, 18, 19]. Papaya leaf curl China virus (PaLCuCNV) is another monopartite begomovirus without satellite DNA, indigenous to China [20]. Both TYLCV and PaLCuCNV can be transmitted by a globally important species of whitefly, provisionally named as Mediterranean (MED), in the B. tabaci complex [21,22,23,24]. Previous studies showed that TYLCV can be more efficiently transmitted by MED whitefly than PaLCuCNV [11, 23, 24]. It seems common that different begomoviruses can be acquired and transmitted by the same whitefly species at different efficiencies, and different whitefly species vary in their capacity in acquiring and transmitting a given begomovirus [9, 23,24,25]. However, up to now, little is yet known about the molecular mechanisms underlying these differences. One way to gain understanding of the molecular mechanisms is to compare the responses of a given whitefly species to different begomoviruses which are transmitted by the whitefly with varied efficiencies, for example MED to TYLCV and PaLCuCNV.

In the past decade, transcriptomics have been used to analyze the interactions between begomoviruses and whiteflies [26,27,28,29]. However, the differently expressed proteins (DEPs) at translational level can better reflect the physiological changes induced by begomoviruses infection. Isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomic approach is a popular methodology in life science, which has been used to investigate the interactions of viruses with various vectors/hosts [30,31,32,33,34].

In this study, we collected un-infected, TYLCV-infected and PaLCuCNV-infected whiteflies respectively, and then used iTRAQ-based quantitative proteomic analysis to elucidate the interactions underlying different combinations of whitefly and begomovirus. Three comparisons were made including TYLCV-infected vs. un-infected, PaLCuCNV-infected vs. un-infected, and TYLCV-infected vs. PaLCuCNV-infected. Our objectives were to provide new visions on the interactions underlying begomovirus transmission by whiteflies and stimulate investigation on these interactions at the proteome level.

Methods

Insects, plants and viruses

The species of whitefly named as MED (mtCOI GenBank accession code: GQ371165) and two tomato cultivars, Solanum lycopersicum cv. Hezuo 903 and cv. Zheza 502, were used for experiments. Clones of TYLCV SH2 (GenBank accession number: AM282874) and PaLCuCNV isolate HeNZM1 (GenBank accession number: FN256260) were obtained from the Institute of Biotechnology, Zhejiang University.

Preparation of whitefly samples

A culture of MED whitefly was reared on un-infected tomato plants (S. lycopersicum cv. Hezuo 903) in insect proof cages. For the experiments, un-infected tomato plants of both Hezuo 903 and Zheza 502 were cultivated to 7–8 true leaf stage when used. For preparation of TYLCV-infected and PaLCuCNV-infected plants, tomato of Hezuo 903 were first cultivated to 3–4 true leaf stage when virus inoculation was conducted, and then the virus-inoculated plants were further cultivated to 7–8 true leaf stage when used. The status of virus-infection of these plants with typical symptoms was verified by PCR detection, and the primers used here are listed in Additional file 1: Table S1.

To obtain whiteflies feeding on un-infected, TYLCV-infected and PaLCuCNV-infected plants, whitefly adults from the MED culture were collected 5–7 d post emergence, and then were placed on un-infected, TYLCV-infected and PaLCuCNV-infected tomato plants of Hezuo 903 respectively to feed for 2 d. The whitefly adults of each of the three treatments were then transferred to feed on un-infected tomato plants of a begomovirus-resistant cultivar (S. lycopersicum cv. Zheza 502) [35] for a further 2 d to reduce/eliminate effects of host plant differences on whiteflies during the 2d treatments (Fig. 1a). The virus-infection status of viruliferous whiteflies or no-viruliferous whiteflies was verified using PCR detection, and the primers used are listed in Additional file 1: Table S1. At this time, adults were collected for proteomics analysis (Fig. 1b). For each of the three treatments, two biological replicates in two separate cages were conducted. All whitefly cohorts of the three treatments were reared in cages at 25–27 °C, 60 ± 10% relative humidity and 14 h light/10 h darkness.

Fig. 1
figure 1

Workflow illustration. (a) Workflow for obtaining whiteflies used in the treatments. Two cultivars of tomato were used in this study, i.e. S. lycopersicum cv. Hezuo 903 (susceptible) and cv. Zheza 502 (resistant). For each of the three treatments, two biological replicates were conducted. All arrows indicate whitefly transfer. (b) Workflow for iTRAQ analysis.

Full size image

Quantitative iTRAQ-LC-MS/MS proteomics analysis

For iTRAQ analysis, whitefly adults of the three treatments as described above, i.e. whiteflies feeding on un-infected, TYLCV-infected and PaLCuCNV-infected plants, were arranged into three combinations for comparison: (i) un-infected vs. TYLCV-infected, (ii) un-infected vs. PaLCuCNV-infected, and (iii) TYLCV-infected vs. PaLCuCNV-infected. For each of the two treatments in a combination for comparison, 0 .1g sample was taken for protein extraction. The methods and procedures of quantitative proteomics analyses followed those of Zhong et al [33]. Briefly, (i) protein extraction, digestion and iTRAQ labeling, (ii) LC-MS/MS analysis, and (iii) proteomic data analysis (Fig. 1b). The raw MS/MS data was converted into MGF format using ProteoWizard tool msConvert (version 3.0.1), and then peptides were identified by searching the MED transcriptomes. We used a MS/MS data interpretation algorithm within Mascot (version 2.3.02). At least one unique peptide was necessary for an identified protein. Based on the data of protein extraction, the bands and repeatability were qualified, and the total content of protein in each of the treatments was greater than 400 μg.

Differential expression ratios of proteins were analyzed by the automated software IQuant (version 2.2.1). To calculate differential expression ratios, all identified spectra from a protein were used to obtain an average protein ratio relative to the control label (i.e. fold change). Student t-test was used to analyze the differential expressed proteins between two treatments. We used P < 0.05 and the fold change > 1.2-fold or < 0.83 fold as the thresholds to judge the significance of differential expressed proteins. We used coefficient of variation (CV), which is defined as the ratio of the standard deviation (SD) to the mean, to evaluate reproducibility.

Gene ontology, pathway enrichment, cluster analysis and cuticle protein family analysis

The identified proteins were categorized according to their Gene Ontology (GO) annotation (http://www.geneontology.org/). The metabolic pathway analysis of the proteins was conducted according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway Database (http://www.genome.jp/kegg). The cluster analysis was conducted using the software Genesis (version 1.8.1). The cuticle protein family analysis was conducted using CutProtFam (aias.biol.uoa.gr/CutProtFam-Pred/home.php) [36].

RT-qPCR validation

To validate results from iTRAQ analysis, genes encoding DEPs among the three treatments were subjected to the RT-qPCR analysis. Twenty adult whiteflies were collected as a group for analyzing the gene expression of DEPs, and three replicates were set for each treatment. For gene expression analysis, total RNA of whitefly was isolated by TRIzol (Ambion, USA) and reverse transcribed using PrimeScript RT reagent Kit (TaKaRa, Japan) following the manufacturer’s protocol. Quantitative PCR (qPCR) was performed on a CFX96™ Real-Time PCR Detection System (Bio-Rad, USA) with SYBR Premix ExTaq II (Takara, Japan). β-actin was used as internal reference, relative abundance of begomovirus or transcripts was calculated by 2-ΔCt. Primers used for real-time PCR are listed in Additional file 1: Table S1.

Results

Basic quantitative parameters

Among the 296,454 spectra generated, 46,699 spectra were identified with 42,153 being unique, 13,042 peptides were identified with 11,954 being unique as judged using 1% PSM (Peptide-spectrum matches) FDR (false discovery rate) (spectrum level), and 3555 proteins were identified with the 1% protein FDR protein levels. In the iTRAQ data, the values of CV exhibit centralized distributions within 0–10% (Additional file 2: Figure S1), indicating a fine reproducibility.

Differentially expressed proteins (DEPs)

For TYLCV-infected vs. un-infected whiteflies, 259 DEPs were identified with 182 being up-regulated and 77 down-regulated (Fig. 2a and b; Additional file 1: Table S2). For PaLCuCNV-infected vs. un-infected, 395 DEPs were identified with 265 being up-regulated and 130 down-regulated (Fig. 2c and d; Additional file 1: Table S3). For TYLCV-infected vs. PaLCuCNV-infected, 74 DEPs were identified with 36 being up-regulated and 38 down-regulated (Fig. 2e and f; Additional file 1: Table S4). Among the 259 DEPs in TYLCV-infected vs. un-infected, 93 were subcategorized into GO classes. These DEPs were found related to receptor group, protein modification process and other processes. Among the 395 DEPs evaluated in PaLCuCNV-infected vs. un-infected, 155 DEPs were subcategorized into GO classes. Most of them were involved in cell cycle, immune response and catabolic process. Among the 74 DEPs evaluated in TYLCV-infected vs. PaLCuCNV-infected, 29 DEPs were subcategorized into GO classes. Groups related to metabolic process exhibited significant enrichment. GO classes with P < 0.05 in each comparison are shown in Fig. 3. In addition, DEPs in the three comparisons were assigned to the reference pathways in KEGG. As a result, 189, 242 and 99 DEPs in the comparison of TYLCV-infected vs. un-infected, PaLCuCNV vs. un-infected, and TYLCV-infected vs. PaLCuCNV-infected were assigned to the reference pathways in KEGG respectively. Finally, 45, 27, 15 pathways were significantly enriched for whiteflies of TYLCV-infected vs. un-infected, PaLCuCNV-infected vs. un-infected, and TYLCV-infected vs. PaLCuCNV-infected, respectively (P < 0.05) (Fig. 4).

Fig. 2
figure 2

Protein expression patterns of MED whiteflies in response to viral infections. Protein expression patterns in whiteflies of TYLCV-infected vs. un-infected (a, b), PaLCuCNV-infected vs. un-infected (c, d), and TYLCV-infected vs. PaLCuCNV-infected (e, f). The Volcano figures of DEPs (a, c and e) depict volcano plot of log2 fold-change (x-axis) versus -log10 Q value (y-axis, representing the probability that the protein is differentially expressed) in each of the three combination for comparison. P < 0.05 and fold change > 1.2 were set as the significant threshold for differential expression. In each of the three diagrams of (a, c, and e), the red dots indicate significantly up-regulations, and the green dots indicate significant down-regulations, while the black dots indicate no significant changes in regulations

Full size image
Fig. 3
figure 3

Gene ontology analysis. The bar chart shows the distribution of corresponding GO terms (P < 0.05). Different colors represent different GO categories. a TYLCV-infected vs. un-infected. b PaLCuCNV-infected vs. un-infected. c TYLCV-infected vs. PaLCuCNV-infected

Full size image
Fig. 4
figure 4

Pathway enrichment analysis. Rich factor is the ratio of differentially expressed protein number annotated in this pathway term to all protein number annotated. Greater rich factor means greater intensiveness. P value ranges from 0~1, and lower P value means greater intensiveness. Displayed here are enriched pathway terms with P < 0.05. a TYLCV-infected vs. un-infected, b PaLCuCNV-infected vs. un-infected, and c TYLCV-infected vs. PaLCuCNV-infected

Full size image

Cluster analysis

A cluster analysis of DEPs was conducted for the three comparisons (Fig. 5), including DEPs related to viral transport, defense response, cell cycle and other processes. Whiteflies infected with different begomoviruses showed some similar responses, including the pathway of ECM-receptor interactions (Table 1), and the up-regulations of some cuticle proteins (Table 2) and proteins related to defense responses (Table 3). In addition, some responses, such as 20S proteasome subunits, were up-regulated only in the comparison of PaLCuCNV-infected vs. un-infected whiteflies (Table 4).

Fig. 5
figure 5

Hierarchical cluster analysis of DEPs. Fold changes of protein abundance in three combinations of comparison were analyzed using the software Genesis (version 1.8.1)

Full size image
Table 1 DEPs involved in the pathway of ECM-receptor interactions
Full size table
Table 2 Up-regulated cuticle proteins in viruliferous whiteflies
Full size table
Table 3 Up-regulated proteins related to defense responses in viruliferous whiteflies
Full size table
Table 4 Up-regulated 20S proteasome subunits in the comparison of PaLCuCNV-infected vs. un-infected
Full size table

RT-qPCR validation of DEPs identified by proteomics

To validate the iTRAQ data, we tested the expression levels of some selected candidate genes in the three treatments. Figure 6 shows the expression patterns of 7 genes, including ras-like protein 3, dystroglycan, integrin alpha-PS2, laminin subunit alpha, cuticle protein 67, PITH domain-containing protein CG6153, and proteasome subunit beta type-6. Consistent with the iTRAQ data, ras-like protein 3, dystroglycan, integrin alpha-PS2, laminin subunit alpha and PITH domain-containing protein were significantly up-regulated after TYLCV/PaLCuCNV infection compared with un-infected whiteflies. Proteasome subunit beta type-6, a kind of 20S proteasome, was significantly up-regulated in the comparison of PaLCuCNV vs. un-infected whiteflies, a result in agreement with the iTRAQ data. However, no significant change was observed in the mRNA expression level of cuticle protein 67.

Fig. 6
figure 6

qPCR analysis of candidate genes. Values of control group (un-infected whitefly) were all set to 1.0 unit. Error bars represent the standard deviation. Significance is indicated with different letters; Student’s t-test

Full size image

Discussion

Three cellular viral receptors induced by TYLCV and PaLCuCNV infections

Binding of a virus to cellular receptors is the key determinant of the physiological outcome of infection [37]. To infect vector cells, viruses need to gain access to the cell surface and then bind their receptor (s). Some attachment factors are required for allowing viral concentration at the cell surface, and following this primary attachment, the viral interaction with specific receptors permits its internalization. This process often requires more than one receptor [38, 39]. A previous study demonstrated that clathrin-mediated endocytosis was involved in TYLCV transport across the vector midgut wall [10], but the receptors that mediated this process remain unknown. According to our iTRAQ data, the expressions of laminin subunit alpha, dystroglycan and integrin alpha-PS2, three proteins in the pathway of extracellular matrix (ECM)-receptor interactions, were significantly up-regulated by 1.36, 1.35 and 1.45 fold respectively in the comparison of TYLCV-infected vs. un-infected whiteflies. Similarly, in the comparison of PaLCuCNV-infected vs. un-infected whiteflies, these three proteins were significantly up-regulated by 1.41, 1.5, 1.36 fold respectively. Laminin has been stated as a cellular attachment receptor for some animal viruses [40, 41]. Alpha-dystroglycan is a type of important cellular viral receptor [37, 42,43,44]. Many integrin subunits have been reported to be usurped by a number of viral and bacterial pathogens in order to gain entrance into host cells [45,46,47,48]. Then, laminin subunit alpha, dystroglycan and integrin alpha-PS2, and three enriched cell surface receptors identified through our iTRAQ analysis after begomovirus infection, may synergistically regulate the endocytosis of begomovirus infection. The fact that clathrin-mediated endocytosis can affect virus penetration of the vector midgut epidermal cells implies that the receptor-mediated endocytosis was essential for viral transmission.

Cuticle proteins induced by TYLCV and PaLCuCNV infection

According to the proteins of insects that have a complete genome, over 1% of the total proteins are cuticle proteins [49], indicating the importance of cuticle proteins in insect body. In our iTRAQ data, three cuticle proteins, cuticle protein 6 (1.26-fold), cuticle protein 67, isoform A (1.25-fold) and cuticle structural protein PCP16.7 (1.22-fold), were significantly up-regulated in the comparison of TYLCV-infected vs. un-infected whiteflies, and two cuticle proteins, cuticle protein 21 (1.35-fold) and cuticle protein 7 (1.21-fold), were significantly up-regulated in the comparison of PaLCuCNV-infected vs. un-infected whiteflies. No cuticle proteins showed down-regulation in both comparisons. According to the analysis from CutProtFam, cuticle protein 6 and cuticle protein 21 belong to the CPR family and RR-2 subgroup, cuticle protein 67, isoform A belongs to CPF family. However, none of the cuticle protein families or sub-families was identified when referring to cuticle structural protein PCP16.7 and cuticle protein 7. For a persistently transmitted virus, e.g. cereal yellow dwarf virus-Rhopalosiphum padi virus, at least four cuticular proteins are involved in the transmission process by the greenbug aphid, Schizaphis graminum [50]. In addition, rice stripe virus can utilize a hemipteran cuticular protein of the small planthopper, Laodelphax striatellus, to facilitate its survival in the hemolymph [51]. So far, the functions of cuticle proteins in begomoviruses transmission have not been reported. Here, we provide a reference for further studies on, for example, clarification of functions of these cuticle proteins identified from our data in begomovirus transmission.

Similar defense responses induced by TYLCV and PaLCuCNV infection

Although insects lack an adaptive immune system, they possess internal defense mechanisms when facing foreign pathogens [52, 53]. In whiteflies, several components have been reported to play a role in viral response, such as the heat shock protein 70 protein and autophagy pathway [14, 54]. According to our iTRAQ data coupled with GO and pathway analysis, in the comparison of TYLCV-infected vs. un-infected whiteflies, some proteins related to defense response, including putative defense protein 3, PITH domain-containing protein CG6153, heat shock factor-binding protein 1 and multidrug resistance-associated protein 1, were significantly up-regulated by 1.57, 1.32, 1.28, 1.25 fold respectively. Similarly, in the comparison of PaLCuCNV-infected vs. un-infected whiteflies, putative defense protein 3, PITH domain-containing protein CG6153 and apoptosis regulator BAX were significantly up-regulated by 1.66, 1.33, 1.31 fold respectively.

Different defense responses induced by TYLCV and PaLCuCNV infection

A given whitefly species can often acquire and transmit different begomoviruses with varied efficiencies [9]. The different characteristics of TYLCV and PaLCuCNV transmission by MED indicate: (i) following viral acquisition, TYLCV can accumulate in the whitefly but PaLCuCNV is unable to [54], and (ii) PaLCuCNV penetrates through the midgut wall of MED less efficiently than TYLCV, resulting in a lower efficiency of PaLCuCNV transmission by MED [11, 24]. In our iTRAQ data, four 20S proteasome subunits were significantly up-regulated in the comparison of PaLCuCNV-infected vs. un-infected whiteflies, namely proteasome subunit beta type-6 (1.31-fold), proteasome subunit alpha type-7-1 (1.27-fold), proteasome subunit alpha type-3 (1.25-fold), and proteasome subunit alpha type-4 (1.24-fold). Interestingly, no 20S or related proteasome subunits showed significant changes in the comparison of TYLCV-infected vs. un-infected whiteflies. Host proteasome-mediated protein proteolysis is known as a common strategy used by both plants and animals for virus degradation and accumulation [55,56,57]. The proteasomes are large multi-subunit proteinase complexes and exist as particles of 20S and of 26S. The 20S particle of ≈700 kDa is an important component of 26S complex of ≈2000 kDa, which is responsible for the degradation of many cellular proteins as a proteolytic core [58, 59]. Ubiquitin-proteasome system has been reported to limit the quantity of begomovirus in whitefly [60]. Thus, in consideration of the potential antiviral function of 20S proteasome, the up-regulated 20S proteasome subunits in PaLCuCNV-infected vs. un-infected whiteflies may be one important factor that leads to the failure of PaLCuCNV accumulation in whitefly body and in turn the lower level of PaLCuCNV acquisition and transmission by MED whitefly.

Conclusions

Previous studies on the interactions between begomoviruses and whiteflies indicate that PaLCuCNV penetrates through the midgut wall of MED whitefly less efficiently than that of Middle East-Asia Minor 1 (MEAM1) whitefly, resulting in a lower efficiency of PaLCuCNV acquisition and transmission by MED than that by MEAM1 [11]. In view of the circulative journey of begomovirus in whitefly [61], both the poorer ability of PaLCuCNV to bind to the midgut cells in MED whitefly and the more sensitive defense responses in MED whitefly could lead to this difference. The data in this study suggests that MED whiteflies infected by TYLCV and PaLCuCNV share some membrane and transport proteins as well as some defense proteins. However, the changes of 20S proteasome subunits between the comparison of TYLCV-infected vs. un-infected whiteflies and PaLCuCNV-infected vs. un-infected whiteflies were in a completely different way. In the future, dsRNA interference could be used to test the roles of laminin subunit alpha, dystroglycan, integrin alpha-PS2 and cuticle proteins in transmission of TYLCV and PaLCuCNV. Taken together, our findings provide new insight into the interactions between whiteflies and begomoviruses, which will serve to provide a number of putative proteins for future investigation on mechanisms underlying whitefly transmission of begomoviruses at the proteome level.

Abbreviations

CV:

Coefficient of variation

DEPs:

Differentially expressed proteins

FDR:

False discovery rate

GO:

Gene Ontology

iTRAQ:

Isobaric tags for relative and absolute quantification

KEGG:

Kyoto Encyclopedia of Genes and Genomes

LC-MS/MS:

Liquid chromatography-tandem mass spectrometry

MED:

Mediterranean

PaLCuCNV:

Papaya leaf curl China virus

PSM:

Peptide-spectrum matches

SD:

Standard deviation

TYLCV:

Tomato yellow leaf curl virus

References

  1. Navas-Castillo J, Fiallo-Olive E, Sanchez-Campos S. Emerging virus diseases transmitted by whiteflies. Annu Rev Phytopathol. 2011;49:219–48.

    Article  CAS  Google Scholar 

  2. Harrison B, Robinson D. Natural genomic and antigenic variation in whiteflytransmitted geminiviruses (begomoviruses). Annu Rev Phytophathol. 1999;37:369–98.

    Article  CAS  Google Scholar 

  3. Zerbini FM, Briddon RW, Idris A, Martin DP, Moriones E, Navas-Castillo J, Rivera-Bustamante R, Roumagnac P, Varsani A. ICTV virus taxonomy profile: Geminiviridae. J. Gen. Virol. 2017;98:131–3.

    Article  CAS  Google Scholar 

  4. Hogenhout SA, Ammer E, Whitfield AE, Redinbaugh MG. Insect vector interactions with persistently transmitted viruses. Annu Rev Phytopathol. 2008;46:327–59.

    Article  CAS  Google Scholar 

  5. Rosen R, Kanakala S, Kliot A, Pakkianathan BC, Farich BC, Santana-Magal N, Elimelech M, Kontsedalov S, Lebedev G, Cilia M, Ghanim M. Persistent, circulative transmission of begomoviruses by whitefly vector. Curr Opin Virol. 2015;15(1–8).

  6. Brown JK, Czosnek H. Whitefly transmission of plant viruses. Adv Bot Res. 2002;36:65–76.

    Article  Google Scholar 

  7. Ghanim M, Morin S, Czosnek H. Rate of Tomato yellow leaf curl virus translocation in the circulative transmission pathway of its vector, the whitefly Bemisia tabaci. Phytopathol. 2001;91:188–96.

    Article  CAS  Google Scholar 

  8. Harrison BD, Swanson MM, Fargette D. Begomovirus coat protein: serology, variation and functions. Physiol Mol Plant Pathol. 2002;60:257–71.

    Article  CAS  Google Scholar 

  9. Wei J, Zhao JJ, Zhang T, Li FF, Ghanim M, Zhou XP, Ye GY, Liu SS, Wang XW. Specific cells in the primary salivary glands of the whitefly Bemisia tabaci control retention and transmission of begomoviruses. J Virol. 2014;88:13460–8.

    Article  Google Scholar 

  10. Pan LL, Chen QF, Zhao JJ, Guo T, Wang XW, Hariton-Shalev A, Czosnek H, Liu SS. Clathrin-mediated endocytosis is involved in Tomato yellow leaf curl virus transport across the midgut barrier of its whitefly vector. Virology. 2017;502:152–9.

    Article  CAS  Google Scholar 

  11. Guo T, Zhao J, Pan LL, Geng L, Lei T, Wang XW, Liu SS. The level of midgut penetration of two begomoviruses affects their acquisition and transmission by two species of Bemisia tabaci. Virology. 2018;515:66–73.

    Article  CAS  Google Scholar 

  12. Kanakala S, Ghanim M. Implication of the whitefly Bemisia tabaci Cyclophilin B protein in the transmission of Tomato yellow leaf curl virus. Front Plant Sci. 2016;7:1702.

    Article  Google Scholar 

  13. Rana VS, Popli S, Saurav GK, Raina HS, Chaubey R, Ramamurthy VV, Rajagopal R. A Bemisia tabaci midgut protein interacts with begomoviruses and plays a role in virus transmission. Cell Microbiol. 2015;18:663–78.

    Article  Google Scholar 

  14. Götz M, Popovski S, Kollenberg M, Gorovits R, Brown JK, Cicero JM, Czosnek H, Winter S, Ghanim M. Implication of Bemisia tabaci heat shock protein 70 in begomovirus-whitefly interactions. J Virol. 2012;86:13241–52.

    Article  Google Scholar 

  15. Hariton-Shalev A, Iris S, Murad G, Liu SS, Henryk C. The whitefly Bemisia tabaci Knottin-1gene is implicated in regulating the quantity of Tomato yellow leaf curl virus ingested and transmitted by the insect. Viruses. 2016;8:205.

    Article  Google Scholar 

  16. Wang ZZ, Shi M, Huang YC, Wang XW, Stanley D, Chen XX. A peptidoglycan recognition protein acts in whitefly (Bemisia tabaci) immunity and involves in begomovirus acquisition. Sci Rep. 2016;6:37806.

    Article  CAS  Google Scholar 

  17. Wang ZZ, Bing XL, Liu SS, Chen XX. RNA interference of an antimicrobial peptide, Btdef, reduces Tomato yellow leaf curl china virus accumulation in the whitefly Bemisia tabaci. Pest Manag Sci. 2017;73:1421–7.

    Article  CAS  Google Scholar 

  18. Czosnek H. Tomato yellow leaf curl virus disease, management, molecular biology, breeding for resistance. Berlin: Springer Netherlands Press; 2007.

    Book  Google Scholar 

  19. Brown JK, Zerbini FM, Navas-Castillo J, Moriones E, Ramos-Sobrinho R, Silva JCF, Fiallo-Olive E, Briddon RW, Hernández-Zepeda C, Idris A, Malathi VG, Martin DP, Rivera-Bustamante R, Ueda S, Varsani A. Revision of Begomovirus, taxonomy based on pairwise sequence comparisons. Arch Virol. 2015;160:1593–619.

    Article  CAS  Google Scholar 

  20. Cai JH, Qin BX, Xie Y, Chen YH. The occurrence of Papaya leaf curl China virus in Nanning and the preliminary study of its midst hosts and transmission by B. tabaci. Plant Prot. 2007;33:57–9. In Chinese with English abstract.

    CAS  Google Scholar 

  21. Hu J, De BP, Zhao H, Wang J, Nardi F, Liu SS. An extensive field survey combined with a phylogenetic analysis reveals rapid and widespread invasion of two alien whiteflies in China. PLoS One. 2011;6:e16061.

    Article  CAS  Google Scholar 

  22. Pan H, Chu D, Ge D, Wang S, Wu Q, Xie W, Jiao X, Liu B, Yang X, Yang N, Su Q, Xu B, Zhang Y. Further spread of and domination by Bemisia tabaci (Hemiptera: Aleyrodidae) biotype Q on field crops in China. J Econ Entomol. 2011;104:978–85.

    Article  Google Scholar 

  23. Li M, Hu J, Xu F, Liu SS. Transmission of Tomato yellow leaf curl virus by two invasive biotypes and a Chinese indigenous biotype of the whitefly Bemisia tabaci. Int J Pest Manage. 2010;56:275–80.

    Article  CAS  Google Scholar 

  24. Guo T, Guo Q, Cui XY, Liu YQ, Hu J, Liu SS. Comparison of transmission of Papaya leaf curl China virus among four cryptic species of the whitefly Bemisia tabaci complex. Sci Rep. 2015;5:15432.

    Article  CAS  Google Scholar 

  25. Jiu M, Zhou XP, Liu SS. Acquisition and transmission of two begomoviruses by the B and a non-B biotype of Bemisia tabaci from Zhejiang, China. J Phytopathol. 2006;154:587–91.

    Article  CAS  Google Scholar 

  26. Sinisterra XH, Mckenzie CL, Hunter WB, Powell CA, Shatters RG. Differential transcriptional activity of plant-pathogenic begomoviruses in their whitefly vector (Bemisia tabaci, Gennadius: Hemiptera Aleyrodidae). J Gen Virol. 2005;86:1525–32.

    Article  CAS  Google Scholar 

  27. Luan JB, Li JM, Varela N, Wang YL, Li FF, Bao YY, Zhang CX, Liu SS, Wang XW. Global analysis of the transcriptional response of whitefly to Tomato yellow leaf curl China virus reveals the relationship of coevolved adaptations. J Virol. 2011;85:3330–40.

    Article  CAS  Google Scholar 

  28. Hasegawa DK, Chen W, Zheng Y, Kaur N, Wintermantel WM, Simmons AM, Fei ZJ, Ling KS. Comparative transcriptome analysis reveals networks of genes activated in the whitefly, Bemisia tabaci when fed on tomato plants infected with Tomato yellow leaf curl virus. Virology. 2018;513:52–64.

    Article  CAS  Google Scholar 

  29. Geng L, Qian LX, Shao RX, Liu YQ, Liu SS, Wang XW. Transcriptome profiling of whitefly guts in response to Tomato yellow leaf curl virus infection. Viro J. 2018;15:14.

    Article  Google Scholar 

  30. Wang H, Wu K, Liu Y, Wu YF, Wang XF. Integrative proteomics to understand the transmission mechanism of barley yellow dwarf virus-GPV by its insect vector Rhopalosiphum padi. Sci Rep. 2015;5:10971.

    Article  Google Scholar 

  31. Wei D, Zeng Y, Xing X, Liu H, Lin M, Han X, Liu X, Liu J. The proteome differences between hepatitis B virus genotype B and genotype C induced hepatocellular carcinoma revealed by iTRAQ based quantitative proteomics. J Proteome Res. 2015;15:487–98.

    Article  Google Scholar 

  32. Lu Q, Bai J, Zhang L, Liu JL, Jiang ZH, Michal JJ, He QD, Jiang P. Two-dimensional liquid chromatography–tandem mass spectrometry coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling approach revealed first proteome profiles of pulmonary alveolar macrophages infected with porcine reproductive and respiratory syndrome virus. J Proteome Res. 2012;11:2890–903.

    Article  CAS  Google Scholar 

  33. Zhong X, Wang ZQ, Xiao RY, Wang YQ, Xie Y, Zhou XP. iTRAQ analysis of the tobacco leaf proteome reveals that RNA-directed DNA methylation (RdDM) has important roles in defense against geminivirus-betasatellite infection. J Proteome. 2016;152:88–101.

    Article  Google Scholar 

  34. Lawrence M, Wynne JW, Kris F, Brian S, Antony B, Michalski WP. Proteomic analysis of Pteropus alecto kidney cells in response to the viral mimic, Poly I: C. Proteome Sci. 2015;13:1–11.

    Article  Google Scholar 

  35. Jiao XY, Zhou XP, Yang YJ, Xie Y. Identification for resistance of tomato varieties against geminiviruses. Acta Phytopathologica Sinica. 2013;43:655–8. In Chinese with English abstract.

    Google Scholar 

  36. Ioannidou ZS, Theodoropoulou MC, Papandreou NC, Willis JH, Hamodrakas SJ. CutProtFam-Pred: Detection and classification of putative structural cuticular proteins from sequence alone, based on profile hidden Markov models. Insect Biochem Mol Biol. 2014;52:51–9.

    Article  CAS  Google Scholar 

  37. Oldstone M, Campbell KP. Decoding arenavirus pathogenesis: essential roles for alpha-dystroglycan-virus interactions and the immune response. Virology. 2011;411:170–9.

    Article  CAS  Google Scholar 

  38. Weigel-Kelley KA, Yoder MC, Srivastava A. 51 integrin as a cellular coreceptor for human parvovirus B19: requirement of functional activation of 1 integrin for viral entry. Blood. 2003;102:3927–33.

    Article  CAS  Google Scholar 

  39. Boulant S, Stanifer M, Pierre-Yves L. Dynamics of Virus-Receptor Interactions in Virus. Binding, Signaling, and Endocytosis. Viruses. 2015;7:2794–815.

    Article  CAS  Google Scholar 

  40. Chen J, He WR, Shen L, Dong H, Yu J, Wang X, Yu S, Li Y, Li S, Luo Y, Sun Y, Qiu HJ. The laminin receptor is a cellular attachment receptor for Classical swine fever virus. J. Virol. 2015;89:4894–906.

    Article  CAS  Google Scholar 

  41. Liu WJ, Li YC, Kou GH, Lo CF. Laminin receptor in shrimp is a cellular attachment receptor for White spot syndrome virus. PLoS ONE. 2016;11:e0156375.

    Article  Google Scholar 

  42. Kunz S, Campbell KP, Oldstone MB. A. α-Dystroglycan can mediate arenavirus infection in the absence of β-dystroglycan. Virology. 2003;316:213–20.

    Article  CAS  Google Scholar 

  43. Kunz S, Rojek JM, Kanagawa M, Spiropoulou CF, Barresi R, Campbell KP, Oldstone MBA. Posttranslational modification of α-Dystroglycan, the cellular receptor for arenaviruses, by the glycosyltransferase large is critical for virus binding. J. Virol. 2005;79:14282–96.

    Article  CAS  Google Scholar 

  44. Shimojima M, Ströher U, Ebihara H, Feldmann H, Kawaoka Y. Identification of cell surface molecules involved in dystroglycan-independent lassa virus cell entry. J Virol. 2012;86:2067–78.

    Article  CAS  Google Scholar 

  45. Gavrilovskaya IN, Brown EJ, Ginsberg MH, Mackow ER. Cellular entry of hantaviruses which cause hemorrhagic fever with renal syndrome is mediated by β3 integrins. J Virol. 1999;73:3951–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Chu JH, Ng ML. Interaction of west nile virus with αvβ3 integrin mediates virus entry into cells. J Biol Chem. 2004;279:54533–41.

    Article  CAS  Google Scholar 

  47. Chiu CY, Mathias P, Nemerow GR, Stewart PL. Structure of adenovirus complexed with its internalization receptor, αvβ5 integrin. J Virol. 1999;73:6759–68.

    CAS  PubMed  PubMed Central  Google Scholar 

  48. Veesler D, Cupelli K, Burger M, Gräberc R, Stehle T, Johnson JE. Single-particle EM reveals plasticity of interactions between the adenovirus penton base and integrin αvβ3. Proc Natl Acad Sci U S A. 2014;111:8815–9.

    Article  CAS  Google Scholar 

  49. Futahashi R, Okamoto S, Kawasaki H, Zhong YS, Iwanaga M, Mita K, Fujiwara H. Genome-wide identification of cuticular protein genes in the silkworm, Bombyx mori. Insect Biochem Mol Biol. 2008;38:1138–46.

    Article  CAS  Google Scholar 

  50. Cilia M, Tamborindeguy C, Fish T, Howe K, Thannhauser TW, Gray S. Genetics coupled to quantitative intact proteomics links heritable aphid and endosymbiont protein expression to circulative polerovirus transmission. J. Virol. 2011;85:2148–66.

  51. Liu WW, Gray S, HuoY, Li L, Wei TY, Wang XF. Proteomic analysis of interaction between a plant virus and its vector insect reveals new functions of hemipteran cuticular protein. Mol Cell Proteomics 2015; 14: 2229–2242.

  52. Salt G. The cellular defence reactions of insects. Cambridge: Cambridge University Press; 1970.

    Book  Google Scholar 

  53. Zhang MM, Chu Y, Zhao ZW, An CJ. Progress in the molecular mechanisms of the innate immune responses in insects. Acta Entomologica Sinica. 2012;55:1221–9 In Chinese with English abstract.

    CAS  Google Scholar 

  54. Wang LL, Wang XR, Wei XM, Huang H, Wu JX, Chen XX, Liu SS, Wang XW. The autophagy pathway participates in resistance to Tomato yellow leaf curl virus infection in whiteflies. Autophagy. 2016;12:1560–74.

    Article  CAS  Google Scholar 

  55. Camborde L, Planchais S, Tournier V, Jakubiec A, Drugeon G, Lacassagne E, Pflieger S, Chenon M, Jupin I. The ubiquitin-proteasome system regulates the accumulation of Turnip yellow mosaic virus RNA-dependent RNA polymerase during viral infection. Plant Cell. 2010;22:3142–52.

    Article  CAS  Google Scholar 

  56. Reichel C, Beachy RN. Degradation of Tobacco mosaic virus movement protein by the 26S proteasome. J. Virol. 2000;74:3330–7.

    Article  CAS  Google Scholar 

  57. Sahana N, Kaur H, Basavaraj TF, Jain RK, Palukaitis P, Canto T, Praveen S. Inhibition of the host proteasome facilitates Papaya ringspot virus accumulation and proteosomal catalytic activity is modulated by viral factor HcPro. Plos ONE. 2012;7:e52546.

    Article  CAS  Google Scholar 

  58. Heinemeyer W, Fischer M, Krimmer T, Stachon U, Wolf DH. The active sites of the eukaryotic 20S proteasome and their involvement in subunit precursor processing. J Bio Chem. 1997;272:25200–9.

    Article  CAS  Google Scholar 

  59. Chen P, Hochstrasser M. Autocatalytic subunit processing couples active site formation in the 20S proteasome to completion of assembly. Cell. 1996;86:961–72.

    Article  CAS  Google Scholar 

  60. Xia WQ, Liang Y, Liu YQ, Liu SS, Wang XW. Effects of ubiquitin-proteasome system on Tomato yellow leaf curl virus in whitefly Bemisia tabaci (Hemiptera: Aleyrodidae). Acta Entomologica Sinica. 2017;60:1411–9 In Chinese with English abstract.

    Google Scholar 

  61. Czosnek H, Hariton-Shalev A, Sobol I, Gorovits R, Ghanim M. The incredible journey of begomoviruses in their whitefly vector. Viruses. 2017;9:273.

    Article  Google Scholar 

Download references

Acknowledgements

We thanks our lab engineer Gen-Hong Yan for cultivating all the plants, and three undergraduates Xi Chen, Yue-Lin Yao and Hui-Ying Zhu for assistance in maintaining whitefly cultures used in this study.

Funding

Financial support for this study was provided by the National Natural Science Foundation of China (31390421), National Key Research and Development Program (2016YFC1200601) and the University of Greenwich, UK (project leader of the Bill & Melinda Gates Foundation Investment ID OPP1149777).

Availability of data and materials

The datasets used in this study are available from the corresponding author on reasonable request, E-mail: shshliu@zju.edu.cn.

Author information

Author notes

  1. Jing Zhao and Yao Chi contributed equally to this work.

Authors and Affiliations

  1. Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou, 310058, People’s Republic of China

    Jing Zhao, Yao Chi, Xin-Jia Zhang, Teng Lei, Xiao-Wei Wang & Shu-Sheng Liu

Authors
  1. Jing Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yao Chi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xin-Jia Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Teng Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiao-Wei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shu-Sheng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

ZJ and CY designed the study, conducted the sample preparation and statistical analysis, and drafted the manuscript. ZXJ and LT conducted the qPCR analysis and provided assistance in the statistical analysis. WXW contributed to manuscript preparation. LSS contributed to experimental design and manuscript revisions. All authors read and approved the manuscript for submission.

Corresponding author

Correspondence to Shu-Sheng Liu.

Ethics declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Additional files

Additional file 1:

Table S1. Primers used in this study. Table S2. DEPs identified in the comparison of TYLCV-infected vs. un-infected. Table S3. DEPs identified in the comparison of PaLCuCNV-infected vs. un-infected. Table S4. DEPs identified in the comparison of TYLCV-infected vs. PaLCuCNV-infected. (DOCX 61 kb)

Additional file 2:

Figure S1. CV distribution of replicates in each of the three combinations for comparison. X-axis is the deviation between the protein ratio of the repeated samples. Y-axis is the percentage of proteins at a certain angle with given levels of quantified proteins. CV value = SD/mean, the lower the value, the better the replication. (A) CV distributions in the comparison of TYLCV-infected vs. un-infected, (B) CV distributions in the comparison of PaLCuCNV-infected vs. un-infected, (C) CV distributions in the comparison of TYLCV-infected vs. PaLCuCNV-infected. (PDF 199 kb)

Rights and permissions

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhao, J., Chi, Y., Zhang, XJ. et al. Comparative proteomic analysis provides new insight into differential transmission of two begomoviruses by a whitefly. Virol J 16, 32 (2019). https://doi.org/10.1186/s12985-019-1138-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12985-019-1138-4

Keywords

  • Whitefly
  • Begomovirus
  • Virus transmission
  • iTRAQ-based proteomics

Virology Journal

ISSN: 1743-422X