Induction of Glandular Trichomes to Control Bemisia tabaci in Tomato Crops: Modulation by the Natural Enemy Nesidiocoris tenuis.

Whitefly-transmitted viruses are one of the biggest threats to tomato (Solanum lycopersicum) growing worldwide. Strategies based on the introgression of resistance traits from wild relatives are promoted to control tomato pests and diseases. Recently, a trichome-based resistance characterizing the wild species Solanum pimpinellifolium was introgressed into a cultivated tomato. An advanced backcross line (BC5S2) exhibiting the presence of acylsugar-associated type IV trichomes, which are lacking in cultivated tomatoes, was effective at controlling whiteflies (Hemiptera: Aleyrodidae) and limiting the spread of whitefly-transmitted viruses. However, at early growth stages, type IV trichome density and acylsugar production are limited; thus, protection against whiteflies and whitefly-transmitted viruses remains irrelevant. In this work, we demonstrate that young BC5S2 tomato plants feeding-punctured by the zoophytophagous predator Nesidiocoris tenuis (Hemiptera: Miridae) displayed an increase (above 50%) in type IV trichome density. Acylsugar production was consistently increased in N. tenuis-punctured BC5S2 plants, which was more likely associated with upregulated expression of the BCKD-E2 gene related to acylsugar biosynthesis. In addition, the infestation of BC5S2 plants with N. tenuis effectively induced the expression of defensive genes involved in the jasmonic acid signaling pathway, resulting in strong repellence to Bemisia tabaci and attractiveness to N. tenuis. Thus, through preplant release of N. tenuis in tomato nurseries carried out in some integrated pest management programs, type IV trichome-expressing plants can be prepared to control whiteflies and whitefly-transmitted viruses at early growth stages. This study emphasizes the advantage of reinforcing constitutive resistance using defense inducers to guarantee robust protection against pests and transmitted viruses.

Crinivirus Tomato Chlorosis Virus Compromises the Control of Tomato Yellow Leaf Curl Virus in Tomato Plants by the Ty-1 Gene.

Tomato yellow leaf curl disease (TYLCD) causes severe damage to tomato crops in warm regions of the world, and is associated with infections of several whitefly (Bemisia tabaci)-transmitted single-stranded (ss)DNA begomoviruses (genus Begomovirus, family Geminiviridae). The most widespread begomovirus isolates associated with TYLCD are those of the type strain of the Tomato yellow leaf curl virus species, known as Israel (TYLCV-IL). The Ty-1 gene is widely used in commercial tomato cultivars to control TYLCV-IL damage, providing resistance to the virus by restricting viral accumulation and tolerance to TYLCD by inhibiting disease symptoms. However, several reports suggest that TYLCV-IL-like isolates are adapting to the Ty-1 gene and are causes of concern for possibly overcoming the provided control. This is the case with TYLCV-IL IS76-like recombinants that have a small genome fragment acquired by genetic exchange from an isolate of Tomato yellow leaf curl Sardinia virus, another begomovirus species associated with TYLCD. Here we show that TYLCV-IL IS76-like isolates partially break down the TYLCD-tolerance provided by the Ty-1 gene and that virulence differences might exist between isolates. Interestingly, we demonstrate that mixed infections with an isolate of the crinivirus (genus Crinivirus, family Closteroviridae) species Tomato chlorosis virus (ToCV), an ssRNA virus also transmitted by B. tabaci and emerging worldwide in tomato crops, boosts the breakdown of the TYLCD-tolerance provided by the Ty-1 gene either with TYLCV-IL IS76-like or canonical TYLCV-IL isolates. Moreover, we demonstrate the incorporation of the Ty-2 gene in Ty-1-commercial tomatoes to restrict (no virus or virus traces, no symptoms) systemic infections of recombinant TYLCV-IL IS76-like and canonical TYLCV-IL isolates, even in the presence of ToCV infections, which provides more robust and durable control of TYLCD.

Begomovirus Tomato Leaf Curl New Delhi Virus Is Seedborne but Not Seed Transmitted in Melon.

Seed transmission can be of considerable relevance to the dissemination of plant viruses in nature and for their prevalence and perpetuation. Long-distance spread of isolates of the begomovirus species Tomato leaf curl New Delhi virus (genus Begomovirus, family Geminiviridae) has recently occurred from Asia to the Middle East and the Mediterranean Basin. Here, we investigated the possible transmission by melon (Cucumis melo L.) seeds of a tomato leaf curl New Delhi virus (ToLCNDV) isolate of the "Spain" strain widely distributed in the Mediterranean area as an alternative mechanism for long-distance spread. PCR amplification detection of ToLCNDV in floral parts and mature seeds of melon plants reveals that this virus is seedborne. "Seedborne" is defined as the ability of a virus to be carried through seeds, which does not necessarily lead to transmission to the next generation. Treatment with a chemical disinfectant significantly reduced the detectable virus associated with melon seeds, suggesting ToLCNDV contamination of the external portion of the seed coat. Also, when the internal fraction of the mature seed (seed cotyledons + embryo) was analyzed by quantitative PCR amplification, ToLCNDV was detectable at low levels, suggesting the potential for viral contamination or infection of the internal portions of seed. However, grow-out studies conducted with melon progeny plants germinated from mature seeds collected from ToLCNDV-infected plants and evaluated at early (1-leaf) or at late (20-leaf) growth stages did not support the transmission of ToLCNDV from seeds to offspring.

Host Plant Resistance to Bemisia tabaci to Control Damage Caused in Tomato Plants by the Emerging Crinivirus Tomato Chlorosis Virus.

Tomato chlorosis virus (genus Crinivirus, family Closteroviridae) (ToCV) is rapidly emerging, causing increased damage to tomato production worldwide. The virus is transmitted in a semipersistent manner by several whitefly (Hemiptera: Aleyrodidae) species and is expanding its geographical and host ranges associated with the emergence of whiteflies of the Bemisia tabaci complex. Control is based essentially on intensive insecticide applications against the insect vector but is largely ineffective. No virus-resistant or tolerant commercial tomato cultivars are available. Recently, a B. tabaci-resistant tomato line based on the introgression of type IV leaf glandular trichomes and secretion of acylsucroses from the wild tomato Solanum pimpinellifolium was shown to effectively control the spread of tomato yellow leaf curl virus, a begomovirus (genus Begomovirus, family Geminiviridae) persistently transmitted by B. tabaci. As short acquisition and transmission periods are associated to the semipersistent transmission of ToCV, its possible control by means of the B. tabaci-resistant tomato could be compromised. Moreover, if the antixenosis effect of the resistance trait present in those tomato plants results in increased B. tabaci mobility, an increased ToCV spread might even occur. We demonstrated, however, that the use of acylsugar-producing B. tabaci-resistant tomatoes effectively controls ToCV spread compared to a near-isogenic line without type IV trichomes and acylsugar secretion. No increase in the primary ToCV spread is observed, and secondary spread could be reduced significantly decreasing the incidence of this virus. The possible use of host plant resistance to whiteflies to limit spread of ToCV opens up new alternatives for a more effective control of this virus to reduce the damage caused in tomato crops.

Revisiting Seed Transmission of the Type Strain of Tomato yellow leaf curl virus in Tomato Plants.

Isolates of the Tomato yellow leaf curl virus (TYLCV) species (genus Begomovirus, family Geminiviridae) infect tomato crops worldwide, causing severe economic damage. Members of the whitefly Bemisia tabaci sibling species group are the vector of begomoviruses, including TYLCV. However, transmission of isolates of the type strain (Israel [IL]) of TYLCV (TYLCV-IL) by tomato seed has recently been reported based on infections occurring in Korea. Because of the consequences of this finding on the epidemiology and control of the disease caused by TYLCV and on the seed market, it was considered essential to revisit and expand those results to other tomato-growing areas. TYLCV DNA content was detected in tomato and Nicotiana benthamiana seed collected from plants naturally or experimentally infected with TYLCV-IL, supporting its seedborne nature. The TYLCV-IL replication detected in tomato and N. benthamiana flower reproductive organs demonstrated close association of this virus with the seed during maturation. However, the significant reduction of TYLCV DNA load after surface disinfections of tomato seed suggests that most of the virus is located externally, as contaminant of the seed coat. Transmission assays, carried out with seven tomato genotypes and more than 3,000 tomato plants, revealed no evidence of seed transmission from "surface-disinfected" or untreated seed for two Mediterranean isolates of TYLCV-IL. Similar results were also obtained for seed collected from TYLCV-IL-infected N. benthamiana plants. The results support the conclusion that TYLCV-IL is seedborne but is not seed transmitted in tomato or N. benthamiana, suggesting that transmission through seed is not a general property of TYLCV.

A Novel Strain of Tomato Leaf Curl New Delhi Virus Has Spread to the Mediterranean Basin.

Tomato leaf curl New Delhi virus (ToLCNDV) is a whitefly-transmitted bipartite begomovirus (genus Begomovirus, family Geminiviridae) that causes damage to multiple cultivated plant species mainly belonging to the Solanaceae and Cucurbitaceae families. ToLCNDV was limited to Asian countries until 2012, when it was first reported in Spain, causing severe epidemics in cucurbit crops. Here, we show that a genetically-uniform ToLCNDV population is present in Spain, compatible with a recent introduction. Analyses of ToLCNDV isolates reported from other parts of the world indicated that this virus has a highly heterogeneous population genetically with no evident geographical, plant host or year-based phylogenetic groups observed. Isolates emerging in Spain belong to a strain that seems to have evolved by recombination. Isolates of this strain seem adapted to infecting cucurbits, but poorly infect tomatoes.

Infectious cDNA clones of the crinivirus Tomato chlorosis virus are competent for systemic plant infection and whitefly-transmission.

Tomato chlorosis virus (ToCV) (genus Crinivirus, family Closteroviridae) causes important emergent diseases in tomato and other solanaceous crops. ToCV is not transmitted mechanically and is naturally transmitted by whiteflies. The ToCV genome consists of two molecules of linear, positive-sense RNA encapsidated into long flexuous virions. We present the construction of full-length cDNA clones of the ToCV genome (RNA1 and RNA2) fused to the SP6 RNA polymerase promoter and under the control of the CaMV 35S promoter. RNA1 replicated in the absence of RNA2 in Nicotiana benthamiana and tomato protoplasts after inoculation with cDNA-derived in vitro transcripts. Agroinfiltration of RNA1 and RNA2 under the 35S promoter resulted in systemic infection in N. benthamiana plants. In addition, tomato plants were infected by grafting with agroinfected N. benthamiana scions, showing the typical ToCV symptoms. The viral progeny generated in tomato was transmissible by the whitefly Bemisia tabaci.