Yellow Leaf Curl Thailand Virus Research Articles

First report of pepper yellow leaf curl Thailand virus infecting hot chili in central Vietnam and Laos.

Hot chili pepper (Capsicum annuum) cultivation has been on the rise in South East Asia to meet export demands. In Thailand, the top chili exporter in South East Asia, chili production has been severely hampered by pepper yellow leaf curl disease (YLCD) caused by the begomovirus pepper yellow leaf curl Thailand virus (PepYLCThV) (Chiemsombat et al., 2018; Suwor et al., 2021). In the neighbouring countries of Laos and Vietnam, a limited survey of chili fields (200 plants in total) in Savannakhet (Savannakhet University campus, n = 150), Laos and Quang Nam province (Ka Dang commune, Dong Giang district, n = 50), central Vietnam in 2023 led to the finding of eight plants (5 in Laos and 3 in Vietnam) exhibiting YLCD-like symptoms, which included bright yellow color in young leaves and leaf curl and mosaic chlorosis in mature leaves (Fig. S1). Total DNA was extracted from leaves of two symptomatic plants (one from Savannakhet and one from Quang Nam) using a cetyltrimethylammonium bromide-based DNA extraction protocol (Doyle & Doyle, 1987; Nguyen et al., 2023). Next, PCR were performed using newly designed PepYLCThV-specific primers based on PepYLCThV sequences in GenBank (Table 1). PCR products of expected sizes were observed in samples with disease symptoms, but not from DNA extracted from C. annuum (cv. VA.99999) grown at the Institute of Biotechnology in Thua Thien Hue, Vietnam (Fig. S2). The amplicons were Sanger sequenced (Apical Scientific, Selangor, Malaysia) and the complete bipartite genome sequence of two isolates ('Sava01' from Laos and 'QNam01' from Vietnam) were obtained. The sequences of the DNA-A component from isolates 'Sava01' (GenBank PP437580) and 'QNam01' (GenBank PP437581) exhibited the highest sequence identity of 99.2% and 94.7% with the PepYLCThV isolate 'ChiangDaoS1' (GenBank OM677627), respectively (Table 2). Conversely, the sequences of the DNA-B component from the isolates 'Sava01' (GenBank PP437579) and 'QNam01' (GenBank PP437582) exhibited the highest similarity of 91.8% and 90.9% with the PepYLCThV isolate 'KKN601' (GenBank MW715820), respectively (Table 2). These results confirmed the presence of PepYLCThV in hot chili pepper plants exhibiting YLCD-like symptoms in central Vietnam and Laos. Infectious clones of PepYLCThV DNA-A and DNA-B (isolate 'QNam01') were created based on the pLX-AS vector as described by Pasin (2022), and transformed into Agrobacterium tumefaciens EHA105. The resulting bacteria were cultured in LB broth containing rifampicin (25 μg/mL) and kanamycin (50 μg/mL) at 28°C and used for agroinoculation of Nicotiana benthamiana (n = 6) and C. annuum (cv. VA.99999, n = 6) (4-6 leaf plants) as described by Pasin (2022). In all N. benthamiana plants, agroinoculation with both DNA-A and DNA-B infectious clones caused stunted growth, severe leaf curl, with yellow and white patches 21 days post inoculation (Fig. S3). In C. annuum plants, symptom expression, which included leaf curl and stunted leaves with yellow mosaic patterns, was observed in two out of six inoculated plants six weeks postinoculation (Fig. S3). PCR assays confirmed the presence of PepYLCThV DNA in N. benthamiana and C. annuum symptomatic leaves (Fig. S4). To our knowledge, this is the first report of pepper yellow leaf curl Thailand virus in hot chili pepper in Laos and central Vietnam. Appropriate containment and management strategies should be developed and implemented to control the spread of this disease in hot chili pepper crops in both countries.

Genetic Diversity and Pathogenicity Characterization of Tomato-Infecting Begomoviruses in Taiwan.

The tomato yellow leaf curl disease (TYLCD) caused by whitefly (Bemisia tabaci)-transmitted begomoviruses (Geminiviridae) has constrained tomato production in Taiwan since 1981. Lisianthus enation leaf curl virus (LELCV), tomato leaf curl Taiwan virus (ToLCTV), and tomato yellow leaf curl Thailand virus (TYLCTHV) were the major viruses associated with TYLCD. In 2019 to 2020, we investigated TYLCD throughout Taiwan, with a 10 to 100% incidence on tomato fields. Begomovirus sequences were detected in 321 out of 506 collected samples by PCR with primers PAL1v1978B and PAR1c715H. In 2015 to 2016, 59 out of 99 samples collected in Hualien-Taitung areas were also found to have begomovirus sequences. Based on the analysis of 68 viral genomic sequences, six begomoviruses were identified, including LELCV, ToLCTV, TYLCTHV, tomato leaf curl Hsinchu virus, and two new begomoviruses, tentatively named tomato leaf curl Chiayi virus (ToLCCYV) and tomato leaf curl Nantou virus (ToLCNTV). Various isolates of LELCV and TYLCTHV were grouped into four and two strains, respectively. Recombinants were detected in LELCV-A, -C, and -D, ToLCCYV, ToLCNTV, and TYLCTHV-F. Based on virus-specific detection, the majority of TYLCD-associated viruses were mixed-infected by TYLCTHV-B with TYLCTHV-F, LELCV-A, -B, or -D, and/or ToLCTV. Meanwhile, viral DNA-B was mostly associated with TYLCTHV, and all identified DNA-Bs were highly homologous with previous TYLCTHV DNA-B. The pathogenicity of selected begomoviruses was confirmed through agroinfection and whitefly transmission. All tomato plants carrying Ty-1/3 and Ty-2 resistant genes were infected by all LELCV strains and ToLCCYV, although they appeared symptomless, suggesting these viruses could be managed through the use of the resistance pyramid.

Harnessing Viral Promoters for Targeted RNAi: Engineering Phloem-Specific Resistance Against Tomato Yellow Leaf Curl Thailand Virus.

Phloem-specific promoter efficiently triggers graft-transmissible RNA interference (gtRNAi). We leveraged a phloem-specific promoter derived from the Rice tungro bacilliform virus, optimizing the RNAi mechanism's efficiency and specificity. Here, we detail the construction of phloem-specific promoter-based gtRNAi system and its application through grafting experiments, demonstrating its effectiveness in inducing tomato yellow leaf curl Thailand virus (TYLCHTV) resistance in non-transgenic scions. This strategy presents a practical application for protecting crops against viruses without genetically modifying the entire plant.

Differential Effects of Two Tomato Begomoviruses on the Life History and Feeding Preference of Bemisia tabaci.

Tomato yellow leaf curl disease, caused by a group of closely related tomato yellow leaf curl viruses, is a major threat to tomato cultivation worldwide. These viruses are primarily transmitted by the sweet potato whitefly (Bemisia tabaci) in a persistent-circulative manner, wherein the virus circulates in the body of B. tabaci and infects its tissues. The complex relationship between viruses and whiteflies significantly influences virus transmission, with studies showing varying effects of the former on the life history and feeding preference of the latter. Whether these effects are direct or indirect, and whether they are negative, neutral, or positive, appears to depend on the specific interactions between virus and whitefly species. The tomato yellow leaf curl Thailand virus (TYLCTHV) and the tomato leaf curl Taiwan virus (ToLCTV) are two prevalent begomoviruses in fields in Taiwan. This study examined the direct and indirect effects of TYLCTHV and ToLCTV on the life history traits (longevity, fecundity, nymph survival, and nymph developmental time) and feeding preference of B. tabaci Middle East-Asia Minor 1 (MEAM1). The results revealed that TYLCTHV had no effects on these life history traits or the feeding preference of MEAM1 whiteflies. Although ToLCTV did not directly affect the longevity and fecundity of MEAM1 whiteflies, their fecundity and the nymph developmental time were negatively affected by feeding on ToLCTV-infected plants. In addition, ToLCTV infection also altered the feeding preference of MEAM1 whiteflies. The different effects of virus infection may contribute to the lower prevalence of ToLCTV compared to TYLCTHV in fields in Taiwan.

Seed and pollen transmission of tomato leaf curl New Delhi virus, tomato leaf curl Taiwan virus, and tomato yellow leaf curl Thailand virus in cucumbers and tomatoes.

Understanding the seed-borne nature of plant viruses is essential for developing disease control strategies and is impactful to the seed market. Here, we investigated seed transmissibility of tomato leaf curl New Delhi virus-cucumber isolate (ToLCNDV-CB) and -oriental melon isolate (ToLCNDV-OM) in cucumber and seed transmissibility of tomato leaf curl Taiwan virus (ToLCTV) and tomato yellow leaf curl Thailand virus (TYLCTHV) in tomato. Parent plants were inoculated using agroinfiltration with virus infectious clones, and virus infection was confirmed by PCR with virus-specific primers. ToLCNDV-CB and ToLCNDV-OM were detected in different parts of the female and male flowers and the fruits of cucumbers. ToLCNDV-CB and ToLCNDV-OM were also detected in cucumber seed coats and seedlings with an infection rate higher than 79%. Similar results were observed with ToLCTV and TYLCTHV as they were detected in different parts of the female and male flowers and fruits of three tomato cultivars. ToLCTV and TYLCTHV were also detected in tomato seed coats and seedlings with an infection rate higher than 36%. In addition, pollen-mediated transmission assays of these four begomoviruses were conducted with pollen derived from virus-infected plants to healthy plants. Results showed that ToLCNDV-CB and ToLCNDV-OM were detected in cross-pollinated cucumber progenies with an infection rate higher than 70%. ToLCTV and TYLCTHV were [also] detected in cross-pollinated tomato progenies with an infection rate higher than 77%. Our results indicated that ToLCNDV, ToLCTV, and TYLCTHV can be transmitted via seeds or pollens of cucumber and tomato plants. To our knowledge, this is the first report documenting the pollen-mediated transmission of begomoviruses.

Virus-virus interactions alter the mechanical transmissibility and host range of begomoviruses.

Begomoviruses are mainly transmitted by whiteflies. However, a few begomoviruses can be transmitted mechanically. Mechanical transmissibility affects begomoviral distribution in the field. In this study, two mechanically transmissible begomoviruses, tomato leaf curl New Delhi virus-oriental melon isolate (ToLCNDV-OM) and tomato yellow leaf curl Thailand virus (TYLCTHV), and two nonmechanically transmissible begomoviruses, ToLCNDV-cucumber isolate (ToLCNDV-CB) and tomato leaf curl Taiwan virus (ToLCTV), were used to study the effects of virus-virus interactions on mechanical transmissibility. Nicotiana benthamiana and host plants were coinoculated through mechanical transmission with inoculants derived from plants that were mix-infected or inoculants derived from individually infected plants, and the inoculants were mixed immediately before inoculation. Our results showed that ToLCNDV-CB was mechanically transmitted with ToLCNDV-OM to N. benthamiana, cucumber, and oriental melon, whereas ToLCTV was mechanically transmitted with TYLCTHV to N. benthamiana and tomato. For crossing host range inoculation, ToLCNDV-CB was mechanically transmitted with TYLCTHV to N. benthamiana and its nonhost tomato, while ToLCTV with ToLCNDV-OM was transmitted to N. benthamiana and its nonhost oriental melon. For sequential inoculation, ToLCNDV-CB and ToLCTV were mechanically transmitted to N. benthamiana plants that were either preinfected with ToLCNDV-OM or TYLCTHV. The results of fluorescence resonance energy transfer analyses showed that the nuclear shuttle protein of ToLCNDV-CB (CBNSP) and the coat protein of ToLCTV (TWCP) localized alone to the nucleus. When coexpressed with movement proteins of ToLCNDV-OM or TYLCTHV, CBNSP and TWCP relocalized to both the nucleus and the cellular periphery and interacted with movement proteins. Our findings indicated that virus-virus interactions in mixed infection circumstances could complement the mechanical transmissibility of nonmechanically transmissible begomoviruses and alter their host range. These findings provide new insight into complex virus-virus interactions and will help us to understand the begomoviral distribution and to reevaluate disease management strategies in the field.

Identification of begomoviruses associated with the insect vector Bemisia tabaci and various host plants on Java Island, Indonesia.

Begomoviruses are economically important plant viruses and are transmitted by Bemisia tabaci which is a complex of various cryptic species. However, it is uncertain whether most begomoviruses that infect host plants are transmitted by B. tabaci at a similar rate. We compared the begomovirus profiles that were detected in a total of 37 whitefly populations and 52 host plants on Java Island, Indonesia. Seven begomovirus species were detected in B. tabaci at different rates: pepper yellow leaf curl Indonesia virus (PepYLCIV, 56.8%), tomato yellow leaf curl Kanchanaburi virus (TYLCKaV, 46.0%), tomato leaf curl New Delhi virus (ToLCNDV, 21.6%), squash leaf curl China virus (SLCCNV, 21.6%), ageratum yellow vein China virus (AYVCNV, 2.7%), mungbean yellow mosaic India virus (MYMIV, 2.7%), and okra enation leaf curl virus (OELCuV, 2.7%). The begomoviruses were detected at different rates in three cryptic species of B. tabaci. In addition, six begomovirus species were detected in the various host plants at different rates: PepYLCIV (67.3%), TYLCKaV (53.9%), ToLCNDV (13.5%), MYMIV (11.5%), AYVCNV (3.9%), and Tomato yellow leaf curl Thailand virus (TYLCTHV) (1.9%). By comparing the virus presence between whiteflies and plants, five begomoviruses (AYVCNV, MYMIV, PepYLCIV, ToLCNDV, and TYLCKaV) were detected in both samples, but their sequence similarity was highly variable depending on the begomovirus themselves; TYLCKaV was highest (99.4%-100%) than any other viruses. Our study suggests B. tabaci acquire begomoviruses at different rates from plants. This study provides important information on the potential variation in the begomovirus transmission mechanism.

Incidence and Molecular Identification of Begomoviruses Infecting Tomato and Pepper in Myanmar.

In Myanmar, yellow mosaic and leaf curl diseases caused by whitefly-transmitted begomoviruses are serious problems for vegetables such as tomatoes and peppers. To investigate the incidence of begomoviruses in Myanmar between 2017 and 2019, a field survey of tomato and pepper plants with virus-like symptoms was conducted in the Naypyitaw, Tatkon, and Mohnyin areas of Myanmar. Among the 59 samples subjected to begomovirus detection using polymerase chain reaction, 59.3% were infected with begomoviruses. Complete genome sequences using rolling circle amplification identified five begomovirus species: tomato yellow leaf curl Thailand virus (TYLCTHV), tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), tobacco leaf curl Yunnan virus (TbLCYnV), chili leaf curl Pakistan virus (ChiLCV/PK), and tobacco curly shoot Myanmar virus (TbCSV-[Myanmar]). Excluding the previously reported TYLCTHV, three begomoviruses (ChiLCV/PK, TYLCKaV, and TbLCYnV) were identified in Myanmar for the first time. Based on the 91% demarcation threshold of begomovirus species, TbCSV-[Myanmar] was identified as a new species in this study. Among these, ChiLCV/PK and TbCSV-[Myanmar] were the most predominant in tomato and pepper fields in Myanmar. Identification of begomovirus species may be helpful for predicting the origin of viruses and preventing their spread.

SEvaluation of processing tomato cultivars for resistance against Tomato yellow leaf curl Thailand virus and high yield production

ISHS IX International Scientific and Practical Conference on Biotechnology as an Instrument for Plant Biodiversity Conservation (physiological, biochemical, embryological, genetic and legal aspects) sEvaluation of processing tomato cultivars for resistance against Tomato yellow leaf curl Thailand virus and high yield production

Translation initiation landscape profiling reveals hidden open-reading frames required for the pathogenesis of tomato yellow leaf curl Thailand virus.

Plant viruses with densely packed genomes employ noncanonical translational strategies to increase the coding capacity for viral function. However, the diverse translational strategies used make it challenging to define the full set of viral genes. Here, using tomato yellow leaf curl Thailand virus (TYLCTHV, genus Begomovirus) as a model system, we identified genes beyond the annotated gene sets by experimentally profiling in vivo translation initiation sites (TISs). We found that unanticipated AUG TISs were prevalent and determined that their usage involves alternative transcriptional and/or translational start sites and is associated with flanking mRNA sequences. Specifically, two downstream in-frame TISs were identified in the viral gene AV2. These TISs were conserved in the begomovirus lineage and led to the translation of different protein isoforms localized to cytoplasmic puncta and at the cell periphery, respectively. In addition, we found translational evidence of an unexplored gene, BV2. BV2 is conserved among TYLCTHV isolates and localizes to the endoplasmic reticulum and plasmodesmata. Mutations of AV2 isoforms and BV2 significantly attenuated disease symptoms in tomato (Solanum lycopersicum). In conclusion, our study pinpointing in vivo TISs untangles the coding complexity of a plant viral genome and, more importantly, illustrates the biological significance of the hidden open-reading frames encoding viral factors for pathogenicity.

Fungal F8-Culture Filtrate Induces Tomato Resistance against Tomato Yellow Leaf Curl Thailand Virus

Tomato (Solanum lycopersicum) is an important economic crop worldwide. However, tomato production is jeopardized by the devastating tomato yellow leaf curl disease caused by whitefly-transmitted begomoviruses (WTBs). In this study, we evaluated the efficacy of our previously developed plant antiviral immunity inducer, fungal F8-culture filtrate, on tomato to combat tomato yellow leaf curl Thailand virus (TYLCTHV), the predominant WTB in Taiwan. Our results indicated that F8-culture filtrate treatment induced strong resistance, did not reduce the growth of tomato, and induced prominent resistance against TYLCTHV both in the greenhouse and in the field. Among TYLCTHV-inoculated Yu-Nu tomato grown in the greenhouse, a greater percentage of plants treated with F8-culture filtrate (43–100%) were healthy-looking compared to the H2O control (0–14%). We found that TYLCTHV cannot move systemically only on the F8-culture filtrate pretreated healthy-looking plants. Tracking the expression of phytohormone-mediated immune maker genes revealed that F8-culture filtrate mainly induced salicylic acid-mediated plant immunity. Furthermore, callose depositions and the expression of the pathogen-induced callose synthase gene, POWDERY MILDEW RESISTANT 4 were only strongly induced by TYLCTHV on tomato pretreated with F8-culture filtrate. This study provides an effective way to induce tomato resistance against TYLCTHV.

Vector Transmission of Tomato Yellow Leaf Curl Thailand Virus by the Whitefly Bemisia tabaci: Circulative or Propagative?

Simple SummaryTomato yellow leaf curl viruses cause disease epidemics in tomato crops in tropical and subtropical areas worldwide. The sweet potato whitefly, Bemisia tabaci, is a vector of this group of viruses. This research studied the transmission biology of tomato yellow leaf curl Thailand virus (TYLCTHV) by B. tabaci, including virus-infected tissues, virus translocation, virus replication, and transovarial transmission (i.e., transmission from mother to progeny via ovaries). We discovered that the virus first infects the alimentary gut, then the hemolymph, and finally the salivary glands of the whitefly. The virus did not replicate in the whitefly during infection. In addition, TYLCTHV was detected in only 10% of infected females’ first-generation progeny, but the progeny was unable to cause viral infection of tomato plants; therefore, there was no evidence of transovarial transmission. When combined with the current literature, our results suggest that B. tabaci transmits TYLCTHV in a persistent-circulative mode.Viruses that cause tomato yellow leaf curl disease are part of a group of viruses of the genus Begomovirus, family Geminiviridae. Tomato-infecting begomoviruses cause epidemics in tomato crops in tropical, subtropical, and Mediterranean climates, and they are exclusively transmitted by Bemisia tabaci in the field. The objective of the present study was to examine the transmission biology of the tomato yellow leaf curl Thailand virus (TYLCTHV) by B. tabaci, including virus-infected tissues, virus translocation, virus replication, and transovarial transmission. The results demonstrated that the virus translocates from the alimentary gut to the salivary glands via the hemolymph, without apparent replication when acquired by B. tabaci. Furthermore, the virus was detected in 10% of the first-generation progeny of viruliferous females, but the progeny was unable to cause the viral infection of host plants. There was no evidence of transovarial transmission of TYLCTHV in B. tabaci. When combined with the current literature, our results suggest that B. tabaci transmits TYLCTHV in a persistent-circulative mode. The present study enhances our understanding of virus–vector interaction and the transmission biology of TYLCTHV in B. tabaci.

Evaluation of phenotyping and genotyping for resistance against Tomato yellow leaf curl Thailand virus (TYLCTHV) validated by Ty-2, Ty-3, and Ty-4 genes in tomato

ISHS I International Symposium on Botanical Gardens and Landscapes Evaluation of phenotyping and genotyping for resistance against Tomato yellow leaf curl Thailand virus (TYLCTHV) validated by Ty-2, Ty-3, and Ty-4 genes in tomato

Identification of the determinant of tomato yellow leaf curl Kanchanaburi virus infectivity in tomato

Geminiviruses cause devastating diseases in solanaceous crops, with the bipartite begomoviruses tomato yellow leaf curl Kanchanaburi virus (TYLCKaV) and pepper yellow leaf curl Thailand virus (PYLCThV) major threats in Southeast Asia. To determine the molecular mechanism of geminivirus infection, we constructed infectious clones of TYLCKaV and PYLCThV. Both constructs infected Nicotiana benthamiana, but only TYLCKaV could infect Solanum lycopersicum ‘A39’. A genome-swapping of TYLCKaV with PYLCThV revealed the TYLCKaV-B genome segment as the determinant of TYLCKaV infectivity in tomato. We constructed five geminivirus clones with chimeric TYLCKaV-B and PYLCThV-B genome segments to narrow down the region determining TYLCKaV infectivity in tomato. Only chimeric clones carrying the TYLCKaV intergenic region (IR) showed infectivity in S. lycopersicum ‘A39’, indicating that the IR of TYLCKaV-B is essential for TYLCKaV infectivity in tomato. Our results provide a foundation for elucidating the molecular mechanism of geminivirus infection in plants.

Evaluation of barrier plants for the cultural control of tomato yellow leaf curl disease

Abstract Barrier cropping plays an essential role in controlling insect pests and insect-transmitted diseases in cultural control. It has been proven efficient in suppressing the spread of nonpersistently transmitted viruses. For suppressing the spread of persistently transmitted viruses, barrier cropping is not considered an effective control strategy because barrier plants cannot act as a virus sink to purge the virus in the vector. However, few successful cases of barrier cropping suppressing the spread of persistently transmitted viruses have been reported. The objectives of the present study were to screen candidates (cucumber, okra, Chinese kale, soybean, and corn) for potential barrier plants to control tomato yellow leaf curl Thailand virus (TYLCTHV) and examine whether prefeeding on these plants can reduce the virus titer in its vector, Bemisia tabaci, thus reducing TYLCTHV transmission. The results revealed that nonviruliferous whiteflies preferred cucumber and okra to tomato, whereas viruliferous whiteflies preferred cucumber to tomato. Although prefeeding on cucumber, okra, and Chinese kale did not reduce the titer of TYLCTHV in viruliferous whiteflies, the vector transmission rate decreased after the whiteflies fed on Chinese kale. It implies that planting Chinese kale as a barrier plant for tomato cultivation may reduce the incidence of TYLCTHV. In addition, the preference to cucumber plants may reduce the incidence of whiteflies acquiring TYLCTHV from virus-infected tomato plants and of viruliferous whiteflies inoculating the virus into healthy tomato plants, thereby reducing the disease incidence. Further field trials of barrier cropping using the candidate plants are warranted.

A Novel Source of Resistance to Pepper yellow leaf curl Thailand virus (PepYLCThV) (Begomovirus) in Chile Pepper

Chile pepper (Capsicum annuum L.) is an increasingly important vegetable and spice crop. Among the most devastating chile pepper–infecting viruses, especially in tropical and subtropical regions, are members of the whitefly transmitted Begomovirus, which cause pepper yellow leaf curl (PYLC). An effective PYLC management strategy is the development of resistant cultivars. However, genetic recombination, acquisition of extra DNA components, and synergistic interactions among different begomoviruses have resulted in the rapid emergence of new viruses that can infect new hosts, cause new disease symptoms, and overcome host resistance. In this project, 98 Capsicum entries comprising breeding lines, open pollinated varieties, genebank accessions, and wild species were screened for resistance to strains of Pepper yellow leaf curl Thailand virus (PepYLCThV). We used a randomized complete block design with three replications and 10 plants per replication in field net-houses at two locations (Khon Kaen and Kamphaeng Saen, Thailand) using augmented inoculation by viruliferous whiteflies. Scoring was done at ≈60, 90, and 120 days after inoculation using a standardized 6-point scale (1 = no symptoms to 6 = very severe symptoms), and the average of the scores of 10 plants within each replication was used for analysis. Although no entry was immune to the disease, the breeding line 9852-123 was highly resistant. Several accessions and lines were moderately resistant at both locations, although a high level of variability within these entries was observed. Overall, the disease severity at the Khon Kaen location was greater compared with Kamphaeng Saen, highlighting the importance of multilocation testing for disease resistance. The resistant entry identified here can be used to study gene action and to move resistance genes into well-adapted germplasm.

Engineering Plant Resistance to Tomato Yellow Leaf Curl Thailand Virus Using a Phloem-Specific Promoter Expressing Hairpin RNA.

Transgenic approaches employing RNA interference (RNAi) strategies have been successfully applied to generate desired traits in plants; however, variations between RNAi transgenic siblings and the ability to quickly apply RNAi resistance to diverse cultivars remain challenging. In this study, we assessed the promoter activity of a cauliflower mosaic virus 35S promoter (35S) and a phloem-specific promoter derived from rice tungro bacilliform virus (RTBV) and their efficacy to drive RNAi against the endogenous glutamate-1-semialdehyde aminotransferase gene (GSA) that acts as a RNAi marker, through chlorophyll synthesis inhibition, and against tomato yellow leaf curl Thailand virus (TYLCTHV), a begomovirus (family Geminiviridae) reported to be the prevalent cause of tomato yellow leaf curl disease (TYLCD) in Taiwan. Transgenic Nicotiana benthamiana expressing hairpin RNA of GSA driven by either the 35S or RTBV promoter revealed that RTBV::hpGSA induced stronger silencing along the vein and more uniformed silencing phenotype among its siblings than 35S::hpGSA. Analysis of transgenic N. benthamiana, 35S::hpTYLCTHV, and RTBV::hpTYLCTHV revealed that, although 35S::hpTYLCTHV generated a higher abundance of small RNA than RTBV::hpTYLCTHV, RTBV::hpTYLCTHV transgenic plants conferred better TYLCTHV resistance than 35S::hpTYLCTHV. Grafting of wild-type (WT) scions to TYLCTHV RNAi rootstocks allowed transferable TYLCTHV resistance to the scion. A TYLCTHV-inoculation assay showed that noninfected WT scions were only observed when grafted to RTBV::hpTYLCTHV rootstocks but not 35S::hpTYLCTHV nor WT rootstocks. Together, our findings demonstrate an approach that may be widely applied to efficiently confer TYLCD resistance.

Occurrence and distribution of begomoviruses infecting tomatoes, peppers and cucurbits in Thailand

Incidence of begomoviruses infecting tomatoes, peppers and cucurbits was investigated in major production fields in the north, northeastern, central and southern regions of Thailand between January 2015 and June 2017. Of 585 samples subjected to begomovirus detection by polymerase chain reaction (PCR), 57% were positive for begomovirus infection. Nucleotide sequence analysis of coat protein (CP) and replication initiation protein (Rep) genes identified 15 previously known begomovirus species, 7 of which were reported infecting these crops in Thailand for the first time. A candidate novel bipartite begomovirus designated Melon yellow mosaic virus was also identified in melons from a northeastern province of Thailand. The predominant begomovirus species were different depending on the host plant (Tomato yellow leaf curl Thailand virus [TYLCTHV] for tomato, Pepper yellow leaf curl Thailand virus [PepYLCTHV] for pepper, Tomato leaf curl New Delhi virus [ToLCNDV] for loofah, bitter gourd, cucumber and melon, and Squash leaf curl China virus [SLCCNV] for pumpkin and wax gourd) and location. The update on diversity and distribution of begomoviruses obtained from this study provides useful information for further epidemiological study and support development of effective disease management through selective use of resistant cultivars suitable for particular plants and locations. Additionally, these data can be applied for development of diagnostic assays to specifically detect begomoviruses for continuous monitoring of viral diversity in these economic crops.

Priming of Plant Resistance to Heat Stress and Tomato Yellow Leaf Curl Thailand Virus With Plant-Derived Materials.

Plants are often simultaneously exposed to diverse environmental stresses, and can tune suitable responses to them through hormones. Salicylic acid (SA) and jasmonic acid (JA) signaling pathways are known to enhance resistance against heat stress and tomato yellow leaf curl Thailand virus (TYLCTHV) infection. However, there is limited information regarding alternative natural priming agents against heat stress and viruses. In this study, two plant-derived priming agents, eugenol and anise oil, were tested for their roles in conferring thermotolerance and virus resistance in tomato plants. Under heat stress, the survival rates and average fresh weight were higher in plants treated with eugenol or anise oil than in control plants. These two priming agents were further tested for antiviral activities. After TYLCTHV infection, the disease incidence and relative abundance of TYLCTHV were lower in anise oil- and eugenol-treated plants than in control plants. Further analyses revealed that a few SA, JA, and RNA silencing genes were enhanced in the former. Moreover, SA, JA, and H2O2 contents increased considerably after eugenol and anise oil treatments. Our findings imply that anise oil and eugenol initiated SA- and JA-mediated defenses to promote thermotolerance and antiviral responses of tomato plants.

Detection of Tomato yellow leaf curl Thailand virus transmitted by Bemisia tabaci Asia I in tomato and pepper

Tomato yellow leaf curl Thailand virus (TYLCTHV) has caused serious yield loss of tomato in Thailand for more than three decades. Control of the virus is essential for successful integrated disease management (IDM) strategies. This study investigated the capability of whitefly (Bemisia tabaci) to transmit TYLCTHV on two host plants—tomato and pepper—and sought to detect the presence of viral DNA in the plants and whiteflies after two transmissions. The new TYLCTHV isolate used in this study (SPN-Tom1A) was 2756 nts long with typical characteristics of TYLCTHV DNA-A. Viral DNA-A (WF-SPN Tom2015) also was detected in viruliferous whiteflies; nucleotide sequence revealed 99% identity to TYLCTHV SPN-Tom1A sequence. In duplicated transmission test, vector B. tabaci Asia I was highly efficient in transmitting TYLCTHV from tomato to tomato by 90–100%, followed by the transmission from tomato to pepper by 55–90%. In contrast, the transmission rate of TYLCTHV pepper to tomato seedlings was nil. Tomato plants showed upward cupping and yellow leaves typical of TYLCTHV infection only when TYLCTHV was transmitted from tomato to tomato, but when transmitted to pepper, the infected peppers were symptomless. Bodies of viruliferous whiteflies fed on tomato and pepper were confirmed by PCR for the presence of TYLCTHV at 58.89–80% and 60–89.45%, respectively. Our results thus provide evidence that pepper infected with TYLCTHV is not a source plant for TYLCTHV transmission to tomato.