Solanum Habrochaites Research Articles

Identification of OSCA gene family in Solanum habrochaites and its function analysis under stress

BackgroundOSCA (hyperosmolality-gated calcium-permeable channel) is a calcium permeable cation channel protein that plays an important role in regulating plant signal transduction. It is involved in sensing changes in extracellular osmotic potential and an increase in Ca2+ concentration. S. habrochaites is a good genetic material for crop improvement against cold, late blight, planthopper and other diseases. Till date, there is no report on OSCA in S. habrochaites. Thus, in this study, we performed a genome-wide screen to identify OSCA genes in S. habrochaites and characterized their responses to biotic and abiotic stresses.ResultsA total of 11 ShOSCA genes distributed on 8 chromosomes were identified. Subcellular localization analysis showed that all members of ShOSCA localized on the plasma membrane and contained multiple stress-related cis acting elements. We observed that genome-wide duplication (WGD) occurred in the genetic evolution of ShOSCA5 (Solhab04g250600) and ShOSCA11 (Solhab12g051500). In addition, repeat events play an important role in the expansion of OSCA gene family. OSCA gene family of S. habrochaites used the time lines of expression studies by qRT-PCR, do indicate OSCAs responded to biotic stress (Botrytis cinerea) and abiotic stress (drought, low temperature and abscisic acid (ABA)). Among them, the expression of ShOSCAs changed significantly under four stresses. The resistance of silencing ShOSCA3 plants to the four stresses was reduced.ConclusionThis study identified the OSCA gene family of S. habrochaites for the first time and analyzed ShOSCA3 has stronger resistance to low temperature, ABA and Botrytis cinerea stress. This study provides a theoretical basis for clarifying the biological function of OSCA, and lays a foundation for tomato crop improvement.

Heat-shock protein 70-a hub gene-underwent adaptive evolution involved in whitefly-wild tomato interaction.

The whitefly Bemisia tabaci causes severe damage to cultivated tomato plants, but actively avoids the wild tomato Solanum habrochaites. Moreover, the mortality of whitefly increases significantly after feeding with the wild tomato. However, additional experiments are warranted to more carefully elucidate the specific molecular elements underlying the interaction between whitefly and wild tomato. Our results showed that S. habrochaites significantly increases the mortality of whitefly adults and decreases both their fertility and fecundity. In addition, the expression of stress-response genes in whitefly after exposure to S. habrochaites was analyzed using RNA sequencing. Weighted gene co-expression network analysis was conducted to identify the hub genes to determine their potential associations with the mortality of whitefly. These results suggested that the expression of heat-shock protein (HSP), multicopper oxidase, and 2-Oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline (OHCU) decarboxylase genes were induced in whitefly. To validate the gene associations with whitefly mortality, a high-throughput in vivo model system and RNAi-based gene silencing were used. The results revealed that the RNAi-mediated depletion of the HSP gene, which belongs to the HSP70 subfamily, increased the mortality of whitefly. Furthermore, the selection pressure analysis showed that a total of five amino acid sites of positive selection were identified, three of which were located in the nucleotide-binding domain and the other two in the substrate-binding domain. This is the first report on the potential implication of HSPs in whitefly-wild plant interactions. This study could more precisely identify the molecular mechanisms of whitefly in response to wild tomatoes. © 2022 Society of Chemical Industry.

Leaf Extracts from Resistant Wild Tomato Can Be Used to Control Late Blight (Phytophthora infestans) in the Cultivated Tomato.

Late blight disease, caused by Phytophthora infestans (Mont.) de Bary, is one of the most challenging diseases threatening tomato production and other Solanaceae crops. Resistance to late blight is found in certain wild species, but the mechanism behind the resistance is not fully understood. The aim of this study was to examine the metabolic profiles in the leaf tissue of late blight-resistant wild tomato and to investigate if leaf extracts from such genotypes could be used to control late blight in tomato production. We included three recognized late blight-resistant wild tomato accessions of Solanum habrochaites (LA1777, LA2855, and LA1352) and two recognized highly susceptible genotypes, S. lycopersicum (‘Super Strain B’) and S. pimpinellifolium (LA0375). The metabolic profiles were obtained in both inoculated and non-inoculated plants by analyzing leaf extracts using high-resolution gas chromatography-mass spectrometry (GC-MS) with three replicate analyses of each genotype. We focused on volatile organic compounds (VOCs) and identified 31 such compounds from the five genotypes with a retention time ranging from 6.6 to 22.8 min. The resistant genotype LA 1777 produced the highest number of VOCs (22 and 21 in the inoculated and control plants, respectively), whereas the susceptible genotype ‘Super Strain B’ produced the lowest number of VOCs (11 and 13 in the respective plants). Among the VOCs, 14 were detected only in the resistant genotypes, while two were detected only in the susceptible ones. In vitro trials, with the use of a detached leaflet assay and whole-plant approach, were conducted. We revealed promising insights regarding late blight management and showed that metabolic profiling may contribute to a better understanding of the mechanisms behind P. infestans resistance in tomato and its wild relatives.

Identification of CDPK Gene Family in Solanum habrochaites and Its Function Analysis under Stress.

Tomato is an important vegetable crop. In the process of tomato production, it will encounter abiotic stress, such as low temperature, drought, and high salt, and biotic stress, such as pathogen infection, which will seriously affect the yield of tomato. Calcium-dependent protein kinase (CDPK) is a class of major calcium signal receptor which has an important regulatory effect on the perception and decoding of calcium signals. CDPK plays a key role in many aspects of plant growth, such as the elongation of pollen tubes, plant growth, and response to biotic and abiotic stress. While some studies have concentrated on Arabidopsis and pepper, Solanum habrochaites is a wild species relative of cultivated tomato and there is no report on CDPK in Solanum habrochaites to date. Using tomato genomic data, this study identified 33 members of the CDPK gene family. Evolutionary analysis divides family members into four Asian groups, of which the CDPK family members have 11 gene replication pairs. Subcellular location analysis showed that most proteins were predicted to be located in the cytoplasm, and less protein existed on the cell membrane. Not all CDPK family members have a transmembrane domain. Cis regulatory elements relating to light, hormones, and drought stress are overrepresented in the promoter region of the CDPK genes in Solanum habrochaites. The expression levels of each gene under biotic stress and abiotic stress were quantified by qRT-PCR. The results showed that members of the CDPK family in Solanum habrochaites respond to different biotic and abiotic stresses. Among them, the expression of ShCDPK6 and ShCDPK26 genes change significantly. ShCDPK6 and ShCDPK26 genes were silenced using VIGS (virus-induced gene silencing), and the silenced plants illustrated reduced stress resistance to Botrytis cinerea, cold, and drought stress. The results of this study will provide a basis for the in-depth study of the CDPK gene family in Solanum habrochaites, laying the foundation for further analysis of the function of the gene family.

Evaluation of responses to tomato brown rugose fruit virus (ToBRFV) and selection of resistant lines in Solanum habrochaites and Solanum peruvianum germplasm

Reactions of plants in 173 wild tomato accessions belonging to Solanum habrochaites and S. peruvianum were studied by inoculation with a tobamovirus, tomato brown rugose fruit virus (ToBRFV). Around 10–50% of plants in nine accessions of S. habrochaites and one of S. peruvianum were demonstrated to be highly resistant. Resistant plants showed no symptoms at 22–24 °C, and no virus could be detected in their inoculated and newly developed leaves using bioassays and RT-qPCR. ToBRFV-resistant plants were also resistant to tobacco mosaic virus and tomato mosaic virus. The susceptible wild tomatoes were infected systemically with ToBRFV showing different severity of symptoms. When resistant plants inoculated with ToBRFV were incubated in a plant growth chamber at a temperature of 33 °C, they expressed mosaic and deformation symptoms, indicating that the resistance was broken at elevated temperature. However, when these plants were transferred to the greenhouse at 24 °C, their newly emerged leaves showed no symptoms, and the virus could not be detected in the new leaves. Cleft grafting was done with scions from a resistant plant of S. habrochaites LA1739 into ToBRFV-infected susceptible tomato rootstock. The scions became infected and showed mosaic symptoms indicating that the resistance was ineffective after grafting. Sequences comparison of Solyc08g075630 loci of nine resistant accessions showed high heterogenity. Only one resistant plant of S. habrochaites carried an allele almost identical to the resistance gene reported previously. All other resistant plants may have probably unknown gene(s) of resistance to ToBRFV.

Identification and Characterization of WRKY41, a Gene Conferring Resistance to Powdery Mildew in Wild Tomato (Solanum habrochaites) LA1777.

WRKYs, a large family of transcription factors, are involved in plant response to biotic and abiotic stresses, but the role of them in tomato resistance to Oidium neolycopersici is still unclear. In this study, we evaluate the role of WRKYs in powdery mildew-resistant wild tomato (Solanum habrochaites) LA1777 defense against O. neolycopersici strain lz (On-lz) using a combination of omics, classical plant pathology- and cell biology-based approaches. A total of 27 WRKYs, belonging to group I, II, and III, were identified as differentially expressed genes in LA1777 against On-lz. It was found that expression of ShWRKY41 was increased after Pseudomonas syringae pv. tomato (Pst) DC3000, On-lz and Botrytis cinerea B05 inoculation or ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) treatment. GUS staining of ShWRKY41 promoter indicated that the expression of ShWRKY41 could be induced by SA and ethylene. Furthermore, ShWRKY41 gene silencing reduced the resistance to On-lz infection by decreasing the generation of H2O2 and HR in LA1777 seedlings. Overall, our research suggests that ShWRKY41 plays a positive role in defense activation and host resistance to O. neolycopersici in wild tomato (S. habrochaites) LA1777.

Distinct Phyllosphere Microbiome of Wild Tomato Species in Central Peru upon Dysbiosis

Plants are colonized by myriads of microbes across kingdoms, which affect host development, fitness, and reproduction. Hence, plant microbiomes have been explored across a broad range of host species, including model organisms, crops, and trees under controlled and natural conditions. Tomato is one of the world’s most important vegetable crops; however, little is known about the microbiota of wild tomato species. To obtain insights into the tomato microbiota occurring in natural environments, we sampled epiphytic microbes from leaves of four tomato species, Solanum habrochaites, S. corneliomulleri, S. peruvianum, and S. pimpinellifolium, from two geographical locations within the Lima region of Peru over 2 consecutive years. Here, a high-throughput sequencing approach was applied to investigate microbial compositions including bacteria, fungi, and eukaryotes across tomato species and geographical locations. The phyllosphere microbiome composition varies between hosts and location. Yet, we identified persistent microbes across tomato species that form the tomato microbial core community. In addition, we phenotypically defined healthy and dysbiotic samples and performed a downstream analysis to reveal the impact on microbial community structures. To do so, we compared microbial diversities, unique OTUs, relative abundances of core taxa, and microbial hub taxa, as well as co-occurrence network characteristics in healthy and dysbiotic tomato leaves and found that dysbiosis affects the phyllosphere microbial composition in a host species-dependent manner. Yet, overall, the present data suggests an enrichment of plant-promoting microbial taxa in healthy leaves, whereas numerous microbial taxa containing plant pathogens occurred in dysbiotic leaves.Concluding, we identify the core phyllosphere microbiome of wild tomato species, and show that the overall phyllosphere microbiome can be impacted by sampling time point, geographical location, host genotype, and plant health. Future studies in these components will help understand the microbial contribution to plant health in natural systems and can be of use in cultivated tomatoes.

Analysis of Salinity Tolerance in Tomato Introgression Lines Based on Morpho-Physiological and Molecular Traits.

The development of salt-tolerant tomato genotypes is a basic requirement to overcome the challenges of tomato production under salinity in the field or soil-free farming. Two groups of eight tomato introgression lines (ILs) each, were evaluated for salinity tolerance. Group-I and the group-II resulted from the following crosses respectively: Solanum lycopersicum cv-6203 × Solanum habrochaites and Solanum lycopersicum M82 × Solanum pennellii. Salt tolerance level was assessed based on a germination percentage under NaCl (0, 75, 100 mM) and in the vegetative stage using a hydroponic growing system (0, 120 mM NaCl). One line from group I (TA1648) and three lines from group II (IL2-1, IL2-3, and IL8-3) were shown to be salt-tolerant since their germination percentages were significantly higher at 75 and 100 mM NaCl than that of their respective cultivated parents cvE6203 and cvM82. Using the hydroponic system, IL TA1648 and IL 2-3 showed the highest value of plant growth traits and chlorophyll concentration. The expression level of eight salt-responsive genes in the leaves and roots of salt-tolerant ILs (TA1648 and IL 2-3) was estimated. Interestingly, SlSOS1, SlNHX2, SlNHX4, and SlERF4 genes were upregulated in leaves of both TA1648 and IL 2-3 genotypes under NaCl stress. While SlHKT1.1, SlNHX2, SlNHX4, and SlERF4 genes were upregulated under salt stress in the roots of both TA1648 and IL 2-3 genotypes. Furthermore, SlSOS2 and SlSOS3 genes were upregulated in TA1648 root and downregulated in IL 2-3. On the contrary, SlSOS1 and SlHKT1.2 genes were upregulated in the IL 2-3 root and downregulated in the TA1648 root. Monitoring of ILs revealed that some of them have inherited salt tolerance from S. habrochaites and S. pennellii genetic background. These ILs can be used in tomato breeding programs to develop salt-tolerant tomatoes or as rootstocks in grafting techniques under saline irrigation conditions.

Characterization of 15-cis-ζ-Carotene Isomerase Z-ISO in Cultivated and Wild Tomato Species Differing in Ripe Fruit Pigmentation.

Isomerization of 9,15,9′-tri-cis-ζ-carotene mediated by 15-cis-ζ-carotene isomerase Z-ISO is a critical step in the biosynthesis of carotenoids, which define fruit color. The tomato clade (Solanum section Lycopersicon) comprises the cultivated tomato (Solanum lycopersicum) and 12 related wild species differing in fruit color and, thus, represents a good model for studying carotenogenesis in fleshy fruit. In this study, we identified homologous Z-ISO genes, including 5′-UTRs and promoter regions, in 12 S. lycopersicum cultivars and 5 wild tomato species (red-fruited Solanum pimpinellifolium, yellow-fruited Solanum cheesmaniae, and green-fruited Solanum chilense, Solanum habrochaites, and Solanum pennellii). Z-ISO homologs had a highly conserved structure, suggesting that Z-ISO performs a similar function in tomato species despite the difference in their fruit color. Z-ISO transcription levels positively correlated with the carotenoid content in ripe fruit of the tomatoes. An analysis of the Z-ISO promoter and 5′-UTR sequences revealed over 130 cis-regulatory elements involved in response to light, stresses, and hormones, and in the binding of transcription factors. Green- and red/yellow-fruited Solanum species differed in the number and position of cis-elements, indicating changes in the transcriptional regulation of Z-ISO expression during tomato evolution, which likely contribute to the difference in fruit color.

Development of biotic stress resistant F1 interspecific hybrid rootstock derived from Solanum lycopersicum and Solanum habrochaites

Tomato is major horticultural plant consumed worldwide. Biotic stress (nematodes, fungus and bacteria) has negative effect on tomato production due to causing reduced yield or plant death. Rootstocks confer resistance to soil-borne pathogen are considered the most effective and environment friendly approach for such a stress management. Thus, development of genetic resources having multiple resistance genes is essential for sustainable tomato breeding. Solanum habrochaites is one of the most studied wild tomato species due to its high genetic potential for biotic and abiotic stresses. In the present study, rootstock potential of an interspecific F1 hybrid derived from S. habrochaites was evaluated as using resistance genes (Frl, I-2, I-3, Mi-3, Pto Ty-1, Ty-3 and Sw-5) specific molecular markers for 6 major tomato diseases and 31 fruit quality traits. The study reported that F1 hybrid had resistance alleles for 5 genes (Frl, I-2, I-3, Pto and Sw-5) confer resistance to fusarium crown rot disease, crown – root rot disease, race 2 and 3 of Fusarium oxysporum f. sp. radicis lycopersici, bacterial speck and tomato spotted wilt virus (TSWV), respectively. Despite high performance of F1 hybrid for biotic stress, the study pointed S. habrochaites specific graft incompatibility due to poor rate of grafting efficiency, small fruit formation and low yield. This is the first comprehensive study evaluated the horticultural performance of an interspecific hybrid in tomato.

Effect of newly developed interspecific hybrid rootstocks on mineral nutrient composition and fruit quality in tomato (Solanum lycopersicum L.)

AbstractTomato (Solanum lycopersicum L.) is one of the important vegetables in the world due to large production area and consumer interest. Biotic and abiotic stresses have negative effect on tomato production. Utilisation of rootstocks conferring resistance to biotic stresses can be considered as the most effective and environment friendly solution in tomato production to overcome this problem. Although wild tomato species is a good rootstock candidate due to its resistance to multiple plant diseases, effects of wild tomato species as rootstock on mineral nutrient composition and fruit quality are not clear. In the present study, effects of interspecific hybrids derived from two wild tomato species (Solanum habrochaites and Solanum penellii) as rootstock on tomato fruit mineral nutrient composition (phosphor (P), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), manganese (Mn), and copper (Cu)) and fruit quality traits (soluble solids content (SSC), pH, percent titratable acidity (TA), and lycopene content) were evaluated. In the study, Amaron, Armstrong, and Arazi commercial rootstocks were used as control and AK0004 F1 (candidate tomato hybrid) was used as a source of scion. As result, only lycopene content was affected by different rootstocks. S. penellii was found to be with more potential for lycopene content. For mineral nutrient composition, all hybrids and controls had similar contents of potassium, phosphor, calcium, and magnesium. Manganese and copper contents decreased in all plants. S. habrochaites had more potential regarding nitrogen, iron, and zinc contens than S. penellii. This is the first study to evaluate two interspecific hybrids derived from S. habrochaites and S. penellii, and the results might be useful to understand effects of rootstocks derived from wild tomato species on mineral nutrient content and fruit quality.

Introgression of the sesquiterpene biosynthesis from Solanum habrochaites to cultivated tomato offers insights into trichome morphology and arthropod resistance

Main conclusionCultivated tomatoes harboring the plastid-derived sesquiterpenes from S. habrochaites have altered type-VI trichome morphology and unveil additional genetic components necessary for piercing-sucking pest resistance.Arthropod resistance in the tomato wild relative Solanum habrochaites LA1777 is linked to specific sesquiterpene biosynthesis. The Sesquiterpene synthase 2 (SsT2) gene cluster on LA1777 chromosome 8 controls plastid-derived sesquiterpene synthesis. The main genes at SsT2 are Z-prenyltransferase (zFPS) and Santalene and Bergamotene Synthase (SBS), which produce α-santalene, β-bergamotene, and α-bergamotene in LA1777 round-shaped type-VI glandular trichomes. Cultivated tomatoes have mushroom-shaped type-VI trichomes with much smaller glands that contain low levels of monoterpenes and cytosolic-derived sesquiterpenes, not presenting the same pest resistance as in LA1777. We successfully transferred zFPS and SBS from LA1777 to cultivated tomato (cv. Micro-Tom, MT) by a backcrossing approach. The trichomes of the MT-Sst2 introgressed line produced high levels of the plastid-derived sesquiterpenes. The type-VI trichome internal storage-cavity size increased in MT-Sst2, probably as an effect of the increased amount of sesquiterpenes, although it was not enough to mimic the round-shaped LA1777 trichomes. The presence of high amounts of plastid-derived sesquiterpenes was also not sufficient to confer resistance to various tomato piercing-sucking pests, indicating that the effect of the sesquiterpenes found in the wild S. habrochaites can be insect specific. Our results provide for a better understanding of the morphology of S. habrochaites type-VI trichomes and paves the way to obtain insect-resistant tomatoes.

Hormone Profiles and Antioxidant Activity of Cultivated and Wild Tomato Seedlings under Low-Temperature Stress

Low temperature is a major limiting factor for the growth and reproduction of some plant species, such as tomato. So far, few studies have been conducted on the effects of low temperature, and the mechanisms of plants’ response to this type of stress is not fully clear. In the current study, the effects of low, nonfreezing temperature (10 °C for three days) on the hormone content, antioxidant activity, and expression patterns of cold-related genes in the leaves of cold-tolerant species (Solanum habrochaites Accession ‘LA1777′) and cold-susceptible species (Solanum lycopersicum cultivar ‘Moneymaker’) were investigated. Low temperature increased the abscisic acid (ABA) content in both tomato species, while the content of zeatin-type cytokinins (ZT) increased in the cold-tolerant species. However, the content of indole-3-acetic acid (IAA) and gibberellic acid (GA) reduced in response to low temperature in susceptible species. Accordingly, cytokinin (CK) is identified as an important hormone associated with low-temperature stress in tomato. In addition, our results indicate that the C-repeat/DRE binding factor 1 (CBF1) gene is less induced in response to low temperature in tomato, although transcription of the inducer of CBF expression 1 (ICE1) gene was upregulated under low temperature in both tomato species. It seems that ICE1 may modulate cold-regulated (COR) genes in a CBF-independent way. In addition, in response to low temperature, the malondialdehyde (MDA) level and membrane stability index (MSI) increased in the susceptible species, indicating that low temperature induces oxidative stress. Additionally, we found that glutathione peroxidase is highly involved in reactive oxygen species (ROS) scavenging induced by low temperature, and antioxidants are more induced in tolerant species. Overall, our results suggest that sub-optimal temperatures promote oxidative stress in tomato and CK is introduced as a factor related to the response to low temperature that requires deeper attention in future breeding programs of tomato.

Migration through a Major Andean Ecogeographic Disruption as a Driver of Genetic and Phenotypic Diversity in a Wild Tomato Species.

Evolutionary dynamics at the population level play a central role in creating the diversity of life on our planet. In this study, we sought to understand the origins of such population-level variation in mating systems and defensive acylsugar chemistry in Solanum habrochaites—a wild tomato species found in diverse Andean habitats in Ecuador and Peru. Using Restriction-site-Associated-DNA-Sequencing (RAD-seq) of 50 S. habrochaites accessions, we identified eight population clusters generated via isolation and hybridization dynamics of 4–6 ancestral populations. Detailed characterization of mating systems of these clusters revealed emergence of multiple self-compatible (SC) groups from progenitor self-incompatible populations in the northern part of the species range. Emergence of these SC groups was also associated with fixation of deleterious alleles inactivating acylsugar acetylation. The Amotape-Huancabamba Zone—a geographical landmark in the Andes with high endemism and isolated microhabitats—was identified as a major driver of differentiation in the northern species range, whereas large geographical distances contributed to population structure and evolution of a novel SC group in the central and southern parts of the range, where the species was also inferred to have originated. Findings presented here highlight the role of the diverse ecogeography of Peru and Ecuador in generating population differentiation, and enhance our understanding of the microevolutionary processes that create biological diversity.

Protein S-nitrosation differentially modulates tomato responses to infection by hemi-biotrophic oomycetes of Phytophthora spp.

Regulation of protein function by reversible S-nitrosation, a post-translational modification based on the attachment of nitroso group to cysteine thiols, has emerged among key mechanisms of NO signalling in plant development and stress responses. S-nitrosoglutathione is regarded as the most abundant low-molecular-weight S-nitrosothiol in plants, where its intracellular concentrations are modulated by S-nitrosoglutathione reductase. We analysed modulations of S-nitrosothiols and protein S-nitrosation mediated by S-nitrosoglutathione reductase in cultivated Solanum lycopersicum (susceptible) and wild Solanum habrochaites (resistant genotype) up to 96 h post inoculation (hpi) by two hemibiotrophic oomycetes, Phytophthora infestans and Phytophthora parasitica. S-nitrosoglutathione reductase activity and protein level were decreased by P. infestans and P. parasitica infection in both genotypes, whereas protein S-nitrosothiols were increased by P. infestans infection, particularly at 72 hpi related to pathogen biotrophy–necrotrophy transition. Increased levels of S-nitrosothiols localised in both proximal and distal parts to the infection site, which suggests together with their localisation to vascular bundles a signalling role in systemic responses. S-nitrosation targets in plants infected with P. infestans identified by a proteomic analysis include namely antioxidant and defence proteins, together with important proteins of metabolic, regulatory and structural functions. Ascorbate peroxidase S-nitrosation was observed in both genotypes in parallel to increased enzyme activity and protein level during P. infestans pathogenesis, namely in the susceptible genotype. These results show important regulatory functions of protein S-nitrosation in concerting molecular mechanisms of plant resistance to hemibiotrophic pathogens.

A single cytochrome P450 oxidase from Solanum habrochaites sequentially oxidizes 7-epi-zingiberene to derivatives toxic to whiteflies and various microorganisms.

Secretions from glandular trichomes potentially protect plants against a variety of aggressors. In the tomato clade of the Solanum genus, glandular trichomes of wild species produce a rich source of chemical diversity at the leaf surface. Previously, 7-epi-zingiberene produced in several accessions of Solanum habrochaites was found to confer resistance to whiteflies (Bemisia tabaci) and other insect pests. Here, we report the identification and characterisation of 9-hydroxy-zingiberene (9HZ) and 9-hydroxy-10,11-epoxyzingiberene (9H10epoZ), two derivatives of 7-epi-zingiberene produced in glandular trichomes of S. habrochaites LA2167. Using a combination of transcriptomics and genetics, we identified a gene coding for a cytochrome P450 oxygenase, ShCYP71D184, that is highly expressed in trichomes and co-segregates with the presence of the zingiberene derivatives. Transient expression assays in Nicotiana benthamiana showed that ShCYP71D184 carries out two successive oxidations to generate 9HZ and 9H10epoZ. Bioactivity assays showed that 9-hydroxy-10,11-epoxyzingiberene in particular exhibits substantial toxicity against B. tabaci and various microorganisms including Phytophthora infestans and Botrytis cinerea. Our work shows that trichome secretions from wild tomato species can provide protection against a wide variety of organisms. In addition, the availability of the genes encoding the enzymes for the pathway of 7-epi-zingiberene derivatives makes it possible to introduce this trait in cultivated tomato by precision breeding.

Dissecting the Genotypic and Environmental Factors Underpinning the Quantitative Trait Variation in a Set of Wild Tomato (Solanum habrochaites LA1777) Introgression Lines

Exotic libraries have proven a powerful tool for the exploitation of wild relatives and quantitative trait loci (QTLs) detection in crop species. In early 2000, an introgression line (IL) population of the wild tomato Solanum habrochaites (SH) acc. LA1777 was developed and made publicly available. Despite the potentiality of the donor parent, so far, these lines have been poorly explored for their agronomic performance and for the identification of genomic regions underlying the variation of quantitative traits (QTLs). Here, we report the evaluation of 19 morpho-agronomic and chemical traits on a set of 39 ILs grown in three consecutive field seasons with the aim to: (a) Determine the overall phenotypic performances of the studied collection, (b) estimate the influence of the genotype (G) and the year of cultivation (Y) and their interaction on the traits analyzed, (c) investigate the plasticity of the traits, and (d) identify whole-genome QTLs in the wild SH background. The ILs showed lower productivity compared to the control genotype, while no major effects were found for the morphological fruit-related traits. Instead, a general increase in the soluble solids content was observed. The combined analysis of G × Y highlighted a major effect of the genotype on trait variation, although yield-related traits were more influenced by environmental factors. In total, 75 associations for 17 traits were detected. Major QTLs increasing soluble solids, pericarp thickness, and trichome density were respectively found on chromosomes 1, 5, and 11 with a percentage variation (PV) of 24.01%, 32.49%, and 200%. Furthermore, different QTLs increasing the color intensity and fruit shape were detected. These results suggest that SH could be a potential source of favorable alleles for qualitative traits despite its inferior phenotype compared to the cultivated parent. The evaluated set of SH LA1777 ILs is a potential for novel allele discovery in wild tomatoes and for breeding purposes towards the exploitation of the available introgressions and for the pyramiding of traits.

Glandular trichome-derived sesquiterpenes of wild tomato accessions (Solanum habrochaites) affect aphid performance and feeding behavior

Glandular trichomes of tomato produce a number of secondary metabolites including terpenes that contribute to host plant resistance against pests. While glandular trichomes of cultivated tomato Solanum lycopersicum primarily accumulate a monoterpene blend, those of wild tomato species like Solanum habrochaites produce various sesquiterpenes. Previous studies have shown that glandular trichome derived terpenes in cultivated and wild tomato species have repellent and toxic activity against multiple biting-chewing herbivores. In contrast, considerably less is known about the effect of these glandular trichome derived terpenes on piercing-sucking herbivores such as aphids. Here, we have screened a collection of S. habrochaites accessions representing five chemotypes that produce distinct sets of sesquiterpenes to identify those affecting the potato aphid (Macrosiphum euphorbiae). Non-choice assays demonstrated that the longevity and fecundity of M. euphorbiae was significantly reduced when kept on the leaf surface of S. habrochaites accessions producing β-caryophyllene and α-humulene, or α-santalene, α-bergamotene, and β-bergamotene, respectively. When M. euphorbiae apterae were feeding on artificial diets with added terpene containing leaf dip extracts, the same β-caryophyllene/α-humulene and α-santalene/α-bergamotene/β-bergamotene producing S. habrochaites accessions were found to affect aphid survivorship and feeding behavior as indicated by gel saliva investment and honeydew production. Olfactometer assays revealed that the sesquiterpenes emitted from these S. habrochaites accessions also have repellent activity against M. euphorbiae alatae affecting their choice behavior prior to landing on host plants. Assays performed with pure sesquiterpene compounds and an introgression line carrying respective S. habrochaites terpene biosynthetic genes in the S. lycopersicum background confirmed that β-caryophyllene/α-humulene and α-santalene/α-bergamotene/β-bergamotene were responsible for the observed effects on performance, feeding and choice behavior of M. euphorbiae.

Selection of F2BC1 tomato genotypes for processing containing high levels of zingiberene and resistant to tomato pinworms

An alternative for the management of the tomato moth is genetic resistance obtained through interspecific crosses. The aim of this study was to select genotypes from the second generation of the first backcrossing (F2BC1) between the cultivar Redencao, which presents processing characteristics, and the wild access Solanum habrochaites var. hirsutum ‘PI-127826’, containing high levels of zingiberene and resistant to tomato moths, using different damage evaluation methodologies. For this purpose, F2BC1 genotypes with different zingiberene levels were submitted to a pest resistance test comprising ten treatments. The treatments consisted of five F2BC1 genotypes with high zingiberene content, three with low content, and the parental plants. The damage caused by the caterpillars were evaluated using two methodologies, a graded visual evaluation and estimates of the percentage of damaged leaf areas. Additionally, population growth was evaluated based on the count of the number of caterpillars present on leaflets. Zingiberene-rich F2BC1 genotypes were less damaged compared to those containing low levels of this substance and the recurrent parent. The two methodologies for assessing damages were demonstrated as feasible. The RVTZ 2011–079-117 and RVTZ 2011–079-335 genotypes presented potential as resistant tomato moth sources.

Salicylic acid treatments induce resistance to Tuta absoluta and Tetranychus urticae on tomato plants

ABSTRACT This study was performed to evaluate the effect of exogenous application of salicylic acid as a resistance inducer to two-spotted spider mite and tomato pinworm on tomato genotypes. Bioassays were carried out in three commercial genotypes (‘Redenção’, ‘Giuliana’, and ‘Alambra’) and one wild genotype (Solanum habrochaites var. hirsutum, accession PI-127826). Average distance travelled by the two-spotted spider mite and number of eggs deposited over the leaflets of the genotypes were investigated. Concerning pinworm bioassay, number of eggs, leafminers on leaflet surface and percentage of leaf area consumed by leafminers were evaluated using a scanner. Furthermore, severity of attack to plants was evaluated by score scale for damages, types of lesions and attacked leaflets. Salicylic acid application induced resistance in the commercial genotypes against both pests, reducing the two-spotted spider mite and pinworm oviposition. The spider mites travelled shorter distance on leaflets of genotypes treated with salicylic acid at intermediate concentrations, indicating a positive effect of substance as inducing resistance. Attack severity by pinworm was also reduced when intermediate doses of salicylic acid were applied. Also, higher resistance was observed on accession PI-127826 as compared to other genotypes. The obtained results indicate that the resistance of PI-127826 was due to constitutive defense, whereas on commercial genotypes defense was dependent on responses mediated by salicylic acid. Hence, salicylic acid treatment, as resistance inducer, seems to be efficient against two-spotted spider mite and pinworm, being a promising tool within an integrated pest management on tomato plants.