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Article

Identification of Begomoviruses from Three Cryptic Species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Nepal

by
Rajendra Acharya
1,
Yam Kumar Shrestha
2,
Mst Fatema Khatun
3 and
Kyeong-Yeoll Lee
1,4,5,*
1
Division of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University, Daegu 41566, Korea
2
Center for Industrial Entomology, Hariharbhawan, Lalitpur 44700, Nepal
3
Department of Entomology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh
4
Institute of Plant Medicine, Kyungpook National University, Daegu 41566, Korea
5
Institute of Quantum Dot Fusion Science and Technology, Kyungpook National University, Gunwi 39061, Korea
*
Author to whom correspondence should be addressed.
Agronomy 2021, 11(10), 2032; https://doi.org/10.3390/agronomy11102032
Submission received: 30 August 2021 / Revised: 1 October 2021 / Accepted: 7 October 2021 / Published: 9 October 2021
(This article belongs to the Special Issue Vector–Pathogen Interactions Affecting Transmission of Plant Pathogens)
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Abstract

:
The Bemisia tabaci species complex consists of at least 44 cryptic species, which are potential vectors of approximately 320 begomovirus species, most of which are significant plant viruses. However, the relationship of begomovirus transmission through vectors at the cryptic species level is uncertain. In our previous study, three cryptic species (Asia I, Asia II 1, and Asia II 5) of B. tabaci were identified from 76 B. tabaci samples collected across 23 districts in Nepal. Using the same individuals we identified seven different begomovirus species (Squash leaf curl China virus [SLCCNV], Tomato leaf curl New Delhi virus [ToLCNDV], Okra enation leaf curl virus [OELCuV], Synedrella leaf curl virus [SyLCV], Tomato leaf curl Kerala virus [ToLCKeV], Ageratum enation virus [AEV], and Tomato leaf curl Karnataka virus [ToLCKV]) by PCR using universal begomovirus primers. The begomoviruses were detected in 55.26% of whitefly samples, and SLCCNV was the most prevalent species (27.63%). Among the three cryptic species of B. tabaci, the virus detection rate was highest in Asia I (60%), followed by Asia II 1 (58.82%) and Asia II 5 (53.06%). Most viruses were detected in all three species, but AEV and ToLCKV were found only in Asia I and Asia II 1, respectively. Geographic analysis showed that SLCCNV was distributed in the whole country, which is similar to the distribution of the Asia II 5 species, but OELCuV and SyLCV were detected only in the middle region of Nepal. Our results provide important information on the begomovirus profile in Nepal which can be beneficial for plant virus risk assessment and develop the management strategies to reduce the damage of whitefly transmitted viruses.

1. Introduction

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a vector of at least 320 plant viruses of the genera Begomovirus, Carlavirus, Crinivirus, Ipomovirus, Polerovirus and Torradovirus [1,2,3,4]. Particularly, B. tabaci is the only vector of Begomovirus, such as the Tomato yellow leaf curl virus (TYLCV), Tomato leaf curl New Delhi virus (ToLCNDV), Squash leaf curl China virus (SLCCNV), and African cassava mosaic virus (ACMV) [5]. These viruses can be transmitted in a persistent, circulative, and non-propagative manner [1,6]. Furthermore, B. tabaci is geographically distributed worldwide with great genetic diversity, forming a species complex consisting of at least 44 morphologically indistinguishable cryptic species [7,8,9,10,11,12,13,14,15]. For example, two cryptic species—Middle East Asia Minor 1 (MEAM1) and Mediterranean (MED)—are highly invasive and cause significant economic damage to various crops worldwide, whereas most other cryptic species are indigenous to specific regions (i.e., Asia and Africa) [16]. In addition, this species has a broad host range of at least 600 different plant species [17]. These characteristics of B. tabaci indicate that this species has significant potential to transmit plant viruses into various crops and spread them widely into most countries across the world.
Previous studies reported the specific interaction between the B. tabaci cryptic species and the transmission rate of begomoviruses [18,19,20]. For example, two cryptic species of B. tabaci—MEAM1 and MED—efficiently transmit the TYLCV, which is highly virulent in the tomato plant [18]. Similarly, the Cotton leaf curl Multan virus (CLCuMuV) transmission by Asia II 1 was higher than those of the cryptic species MEAM1, MED, and Asia I [21]. Bedford et al. [22] and Sánchez-Campos et al. [23] also suggested that different species from the B. tabaci species complex exhibited differential capacities to transmit begomoviruses.
Prevalence and diversity studies of plant viruses are usually based on infected plant samples with virus-like disease symptoms. However, a vector-based plant virus identification technique has been conducted in several studies and provides several advantages [24,25]. Using this technique, which relies on PCR assay of vector insects, researchers can easily detect the viruses and survey their geographic distribution. More importantly, this technique provides information on the relationships between vectors and plant viruses.
Information on the occurrence, distribution, and economic loss of begomovirus in Nepal is limited. Until now, only a few begomoviruses, such as Mungbean yellow vein mosaic Indian virus (MYMIV) [26], Pea leaf distortion virus (PLDV) [27], Ageratum enation virus (AEV) [28] and Ageratum yellow vein virus (AYVV) [29] have been reported in Nepal. Ghimire et al. [30] reported the occurrence of Tomato yellow leaf curl virus (TYLCV) disease in most tomato growing pockets in Nepal and recorded more than 40% yield loss. There are many vegetable and fruit crops showing various begomovirus-like symptoms in Nepal. In addition, B. tabaci which is a vector of begomoviruses is abundant in agricultural fields in Nepal. However, the presence of begomovirus species and their potential vectors are still unknown in Nepal.
In our previous study on the genetic and geographic distribution of B. tabaci species complex in Nepal, we found three cryptic species: Asia I, Asia II 1, and Asia II 5 [31]. We expected that the geographic distribution of begomoviruses that were ingested by different cryptic species of B. tabaci can be determined by analyzing whitefly samples in Nepal. Thus, we analyzed the begomovirus profile in the three cryptic species of B. tabaci in Nepal using a vector-based virus detection technique. This study provides important information on the geographic distribution of begomovirus in Nepal and the potential relationships between B. tabaci cryptic species and begomoviruses.

2. Materials and Methods

2.1. Whitefly Samples

We collected seventy-six B. tabaci samples during the summer of 2017–2019 across 23 districts from east to west Nepal (Table 1) for identification of B. tabaci cryptic species and their geographic distribution, which were reported in our previous study [31]. Samples were collected from different host plants, including vegetables (Abelmoschus esculentus, Cucurbita pepo, Cucumis sativus, Phaseolus vulgaris, and Sechium edule,), oilseed crop (Sesamum indicum), flowers (Bougainvillea spectabilis, Hibiscus rosa-sinensis, Ixora sp., and Salvia officinalis), tuber crop (Smallanthus sonchifolius) and fruit plant (Psidium guajava). All the samples were preserved in ethanol (95%) and stored at −20 °C for further analysis. We used the same individuals of B. tabaci for this study.

2.2. DNA Extraction and Polymerase Chain Reaction

Total genomic DNA was extracted from individual whitefly using a PureLink Genomic DNA Mini Kit (Invitrogen, CA, USA), as described in the commercial kit. A single adult of B. tabaci was placed in a 1.5 mL centrifuge tube containing digestion buffer (180 µL) and homogenized with a plastic homogenizer. By adding 20 µL proteinase K, the sample was incubated at 55 °C for 4 h. Furthermore, DNA was purified using genomic spin column, wash buffer 1 and 2, as described in the kit. Begomovirus acquisition of B. tabaci was determined using universal begomovirus primers Begomo1 (5′CCGTGCTGCTGCCCCCATTGTCCGCGTCAC-3′) and Begomo2 (5′CTGCCACAACCATGGATTCACGCACAGGG-3′) [32]. The PCR product size was around 1100 bp long. For positive and negative controls, we used DNA from the TYLCV-viruliferous B. tabaci and non-viruliferous B. tabaci, respectively, which were reared at Insect Molecular Physiology Laboratory, Kyungpook National University, Daegu, Korea. The total PCR volume of 30 µL contained 15 µL of SolgTM 2x Taq Pre-Mix (Solgent, Daejeon, Korea), 2 µL of each primer (10 pmol/µL), 3 µL of the DNA solution, and 8 µL of distilled water. The mixtures were amplified in a Veriti 96-Well Thermal Cycler (Applied Biosystems, Foster City, CA) with an initial denaturation at 95 °C for 2 min, followed by 35 cycles of 94 °C for 40 s denaturation, 65 °C for 1 min annealing, 72 °C for 30 s extension, and final extension of 7 min at 72 °C. The gel electrophoresis was performed using 1% agarose gel, stained with ethidium bromide solution, and visualized under ultraviolet light. Finally, the amplified PCR products from the gel were purified using the Wizard PCR preps DNA purification system (Promega, Madison, WI, USA).

2.3. Sequence Alignment and Phylogenetic Analyses

Total 42 purified PCR DNA samples were sequenced by using the ABI Prism 3730XL DNA Analyzer (50 cm capillary) (DNA Sequencer) and a BigDye® Terminator v3.1 cycle Sequencing kit (Applied Biosystems) at the Solgent Sequencing Facility (Solgent, Daejeon, Korea). CLUSTAL W [33] was used to align the raw nucleotide sequences, and the resulting alignment was manually edited. Sequences were identified by BLAST searches to the GenBank database of the National Center for Biotechnology Information (NCBI) [34]. Phylogenetic analyses were performed by using the MEGA 6.0 program [35]. The maximum likelihood method was implemented to construct the phylogenetic tree. The robustness of the phylogenetic tree was assessed by 1000 bootstrap replicates [36].

3. Results

3.1. Identification of Begomovirus from Different Cryptic Species of Bemisia tabaci and Their Distribution

Begomoviruses were detected from all three cryptic species sampled in Nepal (Table 1). We identified 7 different begomoviruses: the SLCCNV, ToLCNDV, Okra enation leaf curl virus (OELCuV), Synedrella leaf curl virus (SyLCV), Tomato leaf curl Kerala virus (ToLCKeV), AEV, and Tomato leaf curl Karnataka virus (ToLCKV) from three cryptic species of B. tabaci collected from 18 districts in Nepal (Figure 1 and Figure 2).
Among them, the detection rate of begomoviruses was highest for SLCCNV (27.63%; 21/76) followed by ToLCNDV (10.53%; 8/76), OELCuV (7.89%; 6/76), SyLCV (3.95%; 3/76), and ToLCKeV (2.63%; 2/76). Both AEV and ToLCKV were detected equally at the lowest rate of 1.32% (1/76) (Figure 3). The total detection rate of the begomoviruses in B. tabaci was 55.26% (42/76) (Figure 3). The virus detection rate was similar in three cryptic species such as Asia I (60%; 6/10), Asia II 1 (58.82%; 10/17), and Asia II 5 (53.06%; 26/49). The detection rate of SLCCNV and TOLCNDV were highest in Asia II 5, whereas OELCuV was detected in high frequency in Asia I and Asia II 5. SLCCNV, ToLCNDV, and OELCuV were detected in all three cryptic species, whereas AEV and ToLCKV were found only in Asia I and Asia II 1, respectively (Figure 4).
The geographic distribution of SLCCNV was widespread in Nepal, but both OELCuV and SyLCV were found in the middle region of Nepal, whereas AEV was found in only one location in the Chitwan district located in the central-southern part of Nepal (Figure 1). The nucleotide sequence variations of the seven identified begomoviruses were in the range of 97.10%–99.81% with the related sequences in the GenBank database (Table 1).

3.2. Identification of the Begomoviruses in B. tabaci Collected from Different Host Plants

We collected B. tabaci from 14 plant species: 7 vegetable crops (Abelmoschus esculentus, Cucurbita pepo, Cucumis sativus, Sechium edule, Solanum lycopersicum, Solanum melongena, and Phaseolus vulgaris), 4 flower plants (Hibiscus rosa-sinensis, Ixora sp., Bougainvillea spectabilis, Salvia officinalis), 1 fruit plant (Psidium guajava), 1 oilseed crop (Sesamum indicum), and 1 tuber crop (Smallanthus sonchifolius). The begomoviruses were identified in B. tabaci infesting only in the vegetable crops, but they were not identified in the flower plants, fruit, oilseed, and tuber crop. In addition, SLCCNV was identified in B. tabaci infesting in C. pepo, C. sativus, S. edule, S. lycopersicum, and S. melongena. On the other hand, ToLCNDV was identified in C. sativus, S. lycopersicum, and S. melongena, and OELCuV was identified in A. esculentus, C. pepo, C. sativus, and S. lycopersicum. Finally, AEV and ToLCKV were identified in the B. tabaci samples infesting only S. melongena (Table 2).

4. Discussion

We identified seven different begomoviruses (SLCCNV, ToLCNDV, OELCuV, SyLCV, ToLCKeV, AEV, and ToLCKV) from whitefly samples collected from 18 different locations in Nepal. Among them, only AEV was previously recorded in Nepal [28]. However, the other six begomoviruses were identified for the first time in this study in Nepal. The overall begomovirus detection rate in B. tabaci was 55.26%. While the total percentage of whiteflies containing begomoviruses was similar among the three species, yet the prevalence of the specific begomoviruses differed within the species.
Our results indicated the different geographic distribution of identified begomoviruses. SLCCNV was distributed in the whole country while the other six viruses were distributed in the middle and west regions of Nepal. In comparison with our previous study [31], the begomovirus profile is highly associated with the geographic distribution of three cryptic species of B. tabaci. Particularly, investigation at the eastern region of Nepal showed that distribution of only Asia II 5 which ingested mostly SLCCNV. This result suggests that a strong relationship between begomovirus species and B. tabaci cryptic species.
The bipartite SLCCNV is one of the major begomoviruses in Asian countries, including China, India, Pakistan, Malaysia, Vietnam, and Phillippines in the Cucurbitaceae crops [37,38,39]. Our finding is the first report of SLCCNV presence in Nepal. This virus was detected in the Asia I, Asia II 1, and mostly from Asia II 5 species, which is the most abundant cryptic species of B. tabaci in Nepal [31]. In a previous study, Khatun et al. [40] detected SLCCNV only in the Asia I species. Previously, Dolores and Valdez [39] detected this virus only in squash and pumpkin but not in cucumber and melon. Similarly, Khatun et al. [40] detected this virus in B. tabaci collected from S. melongena and Nicotiana tabacum. However, we detected this virus in B. tabaci collected from C. pepo, C. sativus, S. edule, S. lycopersicum, and S. melongena. This result suggested that the host range of SLCCNV is not restricted only in Cucurbitaceae crops but also poses a potential threat to non-cucurbitaceae crops.
The Tomato leaf curl virus (ToLCV) is one of the most important economic viruses for solanaceous crops in Asia [25,40,41,42,43,44]. In India, 21 different types of ToLCV were recorded [43]. Three types of ToLCV (ToLCNDV, ToLCKeV, and ToLCKV) were found in this study. Among them, ToLCNDV was detected in all three cryptic species from host crops—C. sativus, S. lycopersicum, and S. melongena. Geographically, it was also distributed from the east (Sunsari district) to the west (Banke district) part of Nepal. Our results are similar to those found by Khatun et al. [40]. They also detected ToLCNDV in three cryptic species (Asia I, Asia II 1, and Asia II 5) of B. tabaci that infested P. vulgaris, S. melongena, Dahlia sp., and S. lycopersicum. The ToLCKeV is another important virus infecting solanaceous crops. It is widely distributed in India and Pakistan [25,45]. In Nepal, the ToLCKeV was detected in two cryptic species of B. tabaci—Asia II 1 and Asia II 5—in the host plants C. pepo and C. sativus. We identified it only from the Banke district, which is located very close to India. We suppose that ToLCKeV was disseminated via B. tabaci from India, as it was detected near India. The ToLCKV is also an important virus infecting solanaceous crops and is widely distributed in India and Pakistan [25,46]. We detected ToLCKV in only one sample in Asia II 1 from the host plant S. melongena. We identified it from only one location in Dang, located midwest-southern part of Nepal.
The OELCuV virus was firstly identified in India and later in Pakistan, Bangladesh, Iran, and African countries [40,47,48,49,50,51,52,53,54]. It caused severe disease in okra and an 80% yield loss in India [52]. We detected this virus in the central and western parts of Nepal, including Chitwan, Nawalparasi, Gorkha, and Kathmandu. All three cryptic species of B. tabaci, Asia I, Asia II 1, and Asia II 5 from host plants A. esculentus, C. pepo, C. sativus, S. lycopersicum were infected by this virus in Nepal. However, OELCuV was associated with only Asia I and Asia II 5 in Bangladesh [40]. Hameed et al. [55] reported that the OELCuV is related to the Cotton leaf curl virus, which is more efficiently transmitted by Asia II 1 [56]. Therefore, the OELCuV is a significant potential threat to okra production in Nepal.
The SyLCV is an old-world monopartite begomovirus reported from India, China, and Bangladesh [40]. We found SyLCV from Kaski, Kathmandu, and Lalitpur districts. It was detected in the Asia I and Asia II 1 species of B. tabaci in the host plants C. pepo and S. lycopersicum. In Bangladesh, this virus was found in the species Asia I of B. tabaci in the host plant S. melongena [40]. Therefore, it suggests that SyLCV has a diverse host range and can be acquired by multiple cryptic species of B. tabaci.
The AEV is a monopartite begomovirus and was first identified in Nepal in the late 1990s [28,57]. Currently, it is widely distributed in northern India [28,58,59,60,61] and Pakistan [57]. This virus mainly infects the weed plants under the genus Ageratum and also infects agricultural crops [57]. As we did not collect the B. tabaci samples from weed plants, we found this virus from only one sample collected from Chitwan district, southern Nepal, which is near the border to India. We detected AEV in the Asia I species of B. tabaci collected from the host plant S. melongena, suggesting a new potential host plant, which needs to be validated in the future. Other plants, including S. lycopersicum [62], Carica papaya [63], and Brassica rapa var. rapa [57], were also recorded as the host plants of AEV.
Our results suggest that a vector-based plant virus detection technique provides important information to understand the distribution of begomoviruses. Furthermore, future analysis of plant samples, as well as the characterization of identified viruses such as full genome sequences (DNA A/B) and associated satellites, are required to determine the precise profile of begomoviruses in Nepal.

5. Conclusions

Our study provides the begomovirus profile and their geographic distribution in Nepal using a vector-based PCR analysis at the cryptic species level of B. tabaci. Overall, we detected seven different begomovirus species, including SLCCNV, ToLCNDV, OELCuV, SyLCV, ToLCKeV, AEV, and ToLCKV. SLCCNV was distributed in the whole country while the other six viruses were distributed in the middle and west regions of Nepal. The results of this study can be beneficial for virus disease risk assessment and devising management plans to limit the spread of the whitefly transmitted viruses in Nepal.

Author Contributions

Conceptualization, R.A. and K.-Y.L.; methodology, R.A., Y.K.S. and M.F.K.; validation, Y.K.S. and K.-Y.L.; formal analysis, R.A.; investigation, R.A.; resources, R.A., Y.K.S. and K.-Y.L.; writing—original draft preparation, R.A.; writing—review and editing, R.A., Y.K.S., M.F.K. and K.-Y.L.; supervision, K.-Y.L.; project administration, Y.K.S. and K.-Y.L.; funding acquisition, Y.K.S. and K.-Y.L. All authors have read and agreed to the published version of the manuscript.

Funding

This work was supported by the Research Program for Exportation Support of Agricultural Products, Animal and Plant Quarantine Agency, in the Republic of Korea (Grant #Z-1543086-2017-21-01).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The genetic data presented in this study are publicly available on GenBank, and the accession numbers are reported in Table 1.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. Map of Nepal showing the distribution of the begomoviruses (A) and distribution of Bemisia tabaci cryptic species (B) [31] in the country. The seven different colors represent the detected begomoviruses species from the different cryptic species of B. tabaci (A) whereas different symbols represent B. tabaci cryptic species (B).
Figure 1. Map of Nepal showing the distribution of the begomoviruses (A) and distribution of Bemisia tabaci cryptic species (B) [31] in the country. The seven different colors represent the detected begomoviruses species from the different cryptic species of B. tabaci (A) whereas different symbols represent B. tabaci cryptic species (B).
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Figure 2. Maximum likelihood phylogenetic tree of begomoviruses sequences detected from three different cryptic species of Bemisia tabaci in Nepal. Different colors indicate begomovirus sequences from our samples (42 sequences), and black color indicates reference sequences obtained from the GenBank database (35 sequences). The phylogenetic tree was built in Mega6 using the HKY+G model.
Figure 2. Maximum likelihood phylogenetic tree of begomoviruses sequences detected from three different cryptic species of Bemisia tabaci in Nepal. Different colors indicate begomovirus sequences from our samples (42 sequences), and black color indicates reference sequences obtained from the GenBank database (35 sequences). The phylogenetic tree was built in Mega6 using the HKY+G model.
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Figure 3. Detection rates of different begomoviruses from Bemisia tabaci in Nepal.
Figure 3. Detection rates of different begomoviruses from Bemisia tabaci in Nepal.
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Figure 4. Detection rates of different begomoviruses from three cryptic species of Bemisia tabaci in Nepal.
Figure 4. Detection rates of different begomoviruses from three cryptic species of Bemisia tabaci in Nepal.
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Table
Table 1. Details of Bemisia tabaci samples were collected from different regions in Nepal along with the most identical sequences information from NCBI including their percent identities, numbers of identical bases and accession numbers.
Table 1. Details of Bemisia tabaci samples were collected from different regions in Nepal along with the most identical sequences information from NCBI including their percent identities, numbers of identical bases and accession numbers.
S.N.Samples NameCollection SitesCollection Dates Plant Viruses Detected in B. tabaci
Virus NameAccession Numbers 1% Identity (NCBI)Numbers of Identical BasesAccession Numbers 2
1Ktm-Cuc-10Kritipur, Kathmandu18 July 2017ToLCNDVMZ34513198.741022/1035AY428769
2Lal-Pum-11Lele, Lalitpur19 July 2017SyLCVMZ34512899.611034/1038MK784250
3Ktm-Cuc-13Syuchatar, Kathmandu28 July 2017OELCuVMZ34512299.331037/1044MK784275
4Ktm-Tom-15Syuchatar, Kathmandu28 July 2017OELCuVMZ34512399.331037/1044MK784275
5Ktm-Pum-17Dahachwok, Kathmandu12 August 2017OELCuVMZ34512499.331037/1044MK784275
6Ktm-Tom-18Dahachwok, Kathmandu12 August 2017SyLCVMZ34512999.811036/1038MK784250
7Gor-Pum-19Gorkha, Gorkha13 August 2017OELCuVMZ34512599.331037/1044MK784275
8Kas-Tom-20Pokhara, Kaski13 August 2017SyLCVMZ34513099.611034/1038MK784250
9Naw-Pum-24Daldale, Nawalparasi17 July 2019SLCCNVMZ36287997.981018/1039DQ026296
10Naw-Okr-25Gaidakot, Nawalparasi17 July 2019OELCuVMZ34512699.621039/1043KT390454
11Naw-Pum-26Gaidakot, Nawalparasi17 July 2019SLCCNVMZ36288098.271018/1039DQ026296
12Chi-Okr-27Tandi, Chitwan17 July 2019OELCuVMZ34512799.621040/1044KT390454
13Chi-Pum-28Tandi, Chitwan17 July 2019SLCCNVMZ35719298.071017/1037MN294705
14Chi-Bri-29Ramnagar, Chitwan25 July 2019AEVMZ29146199.421031/1037JX436473
15Gor-Cum-32Manakamana, Gorkha17 July 2019ToLCNDVMZ34513298.651021/1035KM383742
16Gor-Bri-34Ghyalchwok, Gorkha22 May 2019SLCCNVMZ35719398.941026/1037MN294705
17Pyu-Cum-35Khaira, Pyuthan15 August 2019ToLCNDVMZ34513397.971014/1035KM383742
18Dan-Bri-36Ghorahi, Dang23 July 2019ToLCKVMZ29146299.231030/1038LN878125
19Dan-Tom-37Ghorahi, Dang23 July 2019ToLCNDVMZ34513497.11005/1035KM383742
20Dan-Pum-38Ghorahi, Dang23 July 2019SLCCNVMZ35719498.071017/1037MN294705
21Ban-Pum-39Koholpur, Banke23 July 2019ToLCKeVMZ32817398.941031/1042KF551575
22Ban-Cum-40Koholpur, Banke23 July 2019ToLCKeVMZ32817498.941031/1042KF551575
23Ban-Bri-41Koholpur, Banke23 July 2019SLCCNVMZ35719598.361020/1037MT081229
24Kan-Cuc-45Mahendranagar, Kanchanpur24 July 2019SLCCNVMZ35719698.261019/1037MN294705
25Kav-Pum-46Banepa, Kavre28 July 2019SLCCNVMZ35719798.361020/1037MT081229
26Mor-Pum-47Biratnagar, Morang1 August 2019SLCCNVMZ35719898.361020/1037MT081229
27Sun-Pum-49Dharan, Sunsari1 August 2019SLCCNVMZ35719997.691013/1037EU573715
28Sun-Bri-51Dharan, Sunsari1 August 2019ToLCNDVMZ34513598.741022/1035KM383742
29Dhk-Pum-52Guthitar, Dhankuta1 August 2019SLCCNVMZ35720097.781014/1037EU573715
30Ilm-Tom-53Fikkal, Ilam2 August 2019SLCCNVMZ35720197.691013/1037EU573715
31Ilm-Pum-54Fikkal, Ilam2 August 2019SLCCNVMZ35720297.691013/1037EU573715
32Ilm-Bri-55Fikkal, Ilam2 August 2019SLCCNVMZ35720398.261019/1037MN294705
33Pan-Pum-58Lalikharka, Panchthar3 August 2019SLCCNVMZ35720498.261019/1037MN294705
34Jha-Bri-59Kakadvitta, Jhapa3 August 2019SLCCNVMZ35720597.971016/1037MN294705
35Jha-Pum-60Kakadvitta, Jhapa3 August 2019SLCCNVMZ35720697.691013/1037EU573715
36Lal-Tom-64Godamchaur, Lalitpur18 July 2019ToLCNDVMZ34513698.741022/1035AY428769
37Ktm-Cha-67Banasthali, Kathmandu6 June 2019SLCCNVMZ35720798.171018/1037MT081229
38Ktm-Tom-69Naikap, Kathmandu29 July 2019ToLCNDVMZ34513798.841023/1035AY428769
39Ktm-Pum-71New Baneshwor, Kathmandu15 June 2019SLCCNVMZ35720898.461021/1037MT081229
40Ktm-Tom-72Dahachwok, Kathmandu30 July 2019ToLCNDVMZ34513898.841023/1035AY428769
41Bha-Cuc-74Sanga, Bhaktapur28 July 2019SLCCNVMZ35720998.261019/1037MT081229
42Bha-Bri-75Sanga, Bhaktapur28 July 2019SLCCNVMZ35721098.261019/1037MT081229
1 The 6th column refers to the GenBank accession numbers assigned in this work. 2 The 9th column refers to the closest database isolates in the GenBank.
Table
Table 2. List of the begomoviruses detected from the three cryptic species of Bemisia tabaci collected from various host plants in Nepal.
Table 2. List of the begomoviruses detected from the three cryptic species of Bemisia tabaci collected from various host plants in Nepal.
S.N.BegomovirusB. tabaci Cryptic Species
(Number of Virus Detected Samples)		Host Plants for B. tabaci
1Squash leaf curl China virus
(SLCCNV)		Asia I (1), Asia II 1 (2), Asia II 5 (18)Cucurbita pepo, Cucumis sativus, Sechium edule, Solanum lycopersicum, Solanum melongena
2Tomato leaf curl New Delhi virus
(ToLCNDV)		Asia I (1), Asia II 1 (1), Asia II 5 (6)Cucumis sativus, Solanum lycopersicum, Solanum melongena
3Okra enation leaf curl virus
(OELCuV)		Asia I (2), Asia II 1 (3), Asia II 5 (1)Abelmoschus esculentus, Cucurbita pepo, Cucumis sativus, Solanum lycopersicum
4Synedrella leaf curl virus
(SyLCV)		Asia I (1), Asia II 1 (2)Cucurbita pepo, Solanum lycopersicum
5Tomato leaf curl Kerala virus
(ToLCKeV)		Asia II 1 (1), Asia II 5 (1)Cucurbita pepo, Cucumis sativus
6Ageratum enation virus
(AEV)		Asia I (1)Solanum melongena
7Tomato leaf Curl Karnataka virus
(ToLCKV)		Asia II 1 (1)Solanum melongena
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Acharya, R.; Shrestha, Y.K.; Khatun, M.F.; Lee, K.-Y. Identification of Begomoviruses from Three Cryptic Species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Nepal. Agronomy 2021, 11, 2032. https://doi.org/10.3390/agronomy11102032

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Acharya R, Shrestha YK, Khatun MF, Lee K-Y. Identification of Begomoviruses from Three Cryptic Species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Nepal. Agronomy. 2021; 11(10):2032. https://doi.org/10.3390/agronomy11102032

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Acharya, Rajendra, Yam Kumar Shrestha, Mst Fatema Khatun, and Kyeong-Yeoll Lee. 2021. "Identification of Begomoviruses from Three Cryptic Species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Nepal" Agronomy 11, no. 10: 2032. https://doi.org/10.3390/agronomy11102032

APA Style

Acharya, R., Shrestha, Y. K., Khatun, M. F., & Lee, K.-Y. (2021). Identification of Begomoviruses from Three Cryptic Species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) in Nepal. Agronomy, 11(10), 2032. https://doi.org/10.3390/agronomy11102032

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