Genetic Analysis and Combining Ability for Improving Yield and Salt Tolerance in Tomato

Domestication has resulted in reduced salt tolerance in tomato. To identify the genetic components causing this deficiency, we performed a genome-wide association study (GWAS) for root Na+ /K+ ratio in a population consisting of 369 tomato accessions with large natural variations. The most significant variations associated with root Na+ /K+ ratio were identified within the gene SlHAK20 encoding a member of the clade IV HAK/KUP/KT transporters. We further found that SlHAK20 transports Na+ and K+ and regulates Na+ and K+ homeostasis under salt stress conditions. A variation in the coding sequence of SlHAK20 was found to be the causative variant associated with Na+ /K+ ratio and confer salt tolerance in tomato. Knockout mutations in tomato SlHAK20 and the rice homologous genes resulted in hypersensitivity to salt stress. Together, our study uncovered a previously unknown molecular mechanism of salt tolerance responsible for the deficiency in salt tolerance in cultivated tomato varieties. Our findings provide critical information for molecular breeding to improve salt tolerance in tomato and other crops.

Respiratory burst oxidase G (SlRBOHG): A key regulator of H2O2-Mediated Na+ homeostasis and salt tolerance in tomato.

Cell wall proteins perform diverse cellular functions in response to abiotic and biotic stresses. To elucidate the possible mechanisms of salt-stress tolerance in tomato. The 30d seedlings of two tomato genotypes with contrasting salt tolerances were transplanted to salt stress (200 mM NaCl) for three days, and then, the cell wall proteins of seedling roots were analyzed by isobaric tags for relative and absolute quantification (iTRAQ). There were 82 and 81 cell wall proteins that changed significantly in the salt-tolerant tomato IL8-3 and the salt-sensitive tomato M82, respectively. The proteins associated with signal transduction and alterations to cell wall polysaccharides were increased in both IL8-3 and M82 cells wall in response to salt stress. In addition, many different or even opposite metabolic changes occurred between IL8-3 and M82 in response to salt stress. The salt-tolerant tomato IL8-3 experienced not only significantly decreased in Na+ accumulation but also an obviously enhanced in regulating redox balance and cell wall lignification in response to salt stress. Taken together, these results provide novel insight for further understanding the molecular mechanism of salt tolerance in tomato.

Polyamines act as protective compounds directly protecting plants from stress-related damage, while also acting as signaling molecules to participate in serious abiotic stresses. However, the molecular mechanisms underlying these effects are poorly understood. Here, we utilized metabolome genome-wide association study to investigate the polyamine content of wild and cultivated tomato accessions, and we discovered a new gene cluster that drove polyamine content during tomato domestication. The gene cluster contains two polyphenol oxidases (SlPPOE and SlPPOF), two BAHD acyltransferases (SlAT4 and SlAT5), a coumaroyl-CoA ligase (Sl4CL6), and a polyamine uptake transporter (SlPUT3). SlPUT3 mediates polyamine uptake and transport, while the five other genes are involved in polyamine modification. Further salt tolerance assays demonstrated that SlPPOE, SlPPOF, and SlAT5 overexpression lines showed greater phenolamide accumulation and salt tolerance as compared with wild-type (WT). Meanwhile, the exogenous application of Spm to SlPUT3-OE lines displayed salt tolerance compared with WT, while having the opposite effect in slput3 lines, confirms that the polyamine and phenolamide can play a protective role by alleviating cell damage. SlPUT3 interacted with SlPIP2;4, a H2O2 transport protein, to maintain H2O2 homeostasis. Polyamine-derived H2O2 linked Spm to stress responses, suggesting that Spm signaling activates stress response pathways. Collectively, our finding reveals that the H2O2-polyamine-phenolamide module coordinately enhanced tomato salt stress tolerance and provide a foundation for tomato stress-resistance breeding.

The possible use of in vitro shoot morphogenesis and shoot apex culture to evaluate salt tolerance in cultivated tomato (Lycopersicon esculentum Mill.) has been analyzed, using two cultivars with similar salt tolerance, Pera and Hellfrucht fruhstamm (HF). The effect of salt on shoot regeneration was studied by culturing leaf explants on media supplemented with 0, 43, 86, 129 and 172 mM NaCl. The presence of NaCl in the regeneration media at 86 mM strongly inhibited shoot regeneration in the cultivar HF, but not in Pera. However, the substitution of NaCl by mannitol, maintaining the same water potential in the culture media, decreased the regeneration percentage in Pera but did not affect HF. Shoot apices of both cultivars were also subcultured at 6-week intervals, for 4 subcultures, at the same NaCl concentrations as used in the previous experiment, and the shoot growth, leaf and root number, rooted shoot and shoot necrosis were recorded at the end of each subculture. Root formation was the parameter most affected by salt in both cultivars, Pera being more sensitive than HF. The substitution of NaCl by mannitol significantly increased the percentage of rooted shoots in Pera after four subcultures, and slightly decreased this percentage in HF. Shoot necrosis was only observed in the last subculture at NaCl higher than 86 mM, the percentage of necrotic shoots being higher in Pera than in HF (75% and 45%, respectively). The lack of agreement between the results obtained with the in vitro tests, e.g., adventitious shoot formation and growth of apical stem sections, suggests that this approach may not be a reliable tool to evaluate salt tolerance in cultivated tomato.

We have identified a plasma membrane Na(+)/H(+) antiporter gene from tomato (Solanum lycopersicum), SlSOS1, and used heterologous expression in yeast to confirm that SlSOS1 was the functional homolog of AtSOS1. Using post-transcriptional gene silencing, we evaluated the role played by SlSOS1 in long-distance Na(+) transport and salt tolerance of tomato. Tomato was used because of its anatomical structure, more complex than that of Arabidopsis, and its agricultural significance. Transgenic tomato plants with reduced expression of SlSOS1 exhibited reduced growth rate compared to wild-type (WT) plants in saline conditions. This sensitivity correlated with higher accumulation of Na(+) in leaves and roots, but lower contents in stems of silenced plants under salt stress. Differential distribution of Na(+) and lower net Na(+) flux were observed in the xylem sap in the suppressed plants. In addition, K(+) concentration was lower in roots of silenced plants than in WT. Our results demonstrate that SlSOS1 antiporter is not only essential in maintaining ion homeostasis under salinity, but also critical for the partitioning of Na(+) between plant organs. The ability of tomato plants to retain Na(+) in the stems, thus preventing Na(+) from reaching the photosynthetic tissues, is largely dependent on the function of SlSOS1.

Understanding salt tolerance in tomato (Solanum lycopersicum L.) landraces will facilitate their use in genetic improvement. The study assessed the morpho-physiological variability of Hail tomato landraces in response to different salinity levels at seedling stages and recommended a tomato salt-tolerant landrace for future breeding programs. Three tomato landraces, Hail 548, Hail 747, and Hail 1072 were tested under three salinity levels: 75, 150, and 300 mM NaCl. Salinity stress reduced shoots’ fresh and dry weight by 71% and 72%, and roots were 86.5% and 78.6%, respectively. There was 22% reduced chlorophyll content, carotene content by 18.6%, and anthocyanin by 41.1%. Proline content increased for stressed treatments. The 300 mM NaCl treatment recorded the most proline content increases (67.37 mg/g fresh weight), with a percent increase in proline reaching 61.67% in Hail 747. Superoxide dismutase (SOD) activity decreased by 65% in Hail 548, while it relatively increased in Hail 747 and Hail 1072 treated with 300 mM NaCl. Catalase (CAT) activity was enhanced by salt stress in Hail 548 and recorded 7.6%, increasing at 75 and 5.1% at 300 mM NaCl. It revealed a reduction in malondialdehyde (MDA) at the 300 mM NaCl concentration in both Hail 548 and Hail 1072 landraces. Increasing salt concentrations showed a reduction in transpiration rate of 70.55%, 7.13% in stomatal conductance, and 72.34% in photosynthetic rate. K+/Na+ ratios decreased from 56% for 75 mM NaCl to 85% for 300 mM NaCl treatments in all genotypes. The response to salt stress in landraces involved some modifications in morphology, physiology, and metabolism. The landrace Hail 548 may have better protection against salt stress and observed protection against reactive oxygen species (ROS) by increasing enzymatic “antioxidants” activity under salt stress.

Tomato is a globally important horticultural and economic crop, but its productivity is severely affected by various stresses. Plant small secretory peptides have been identified as crucial mediators in plant resistance. Here, we conducted a comparative transcriptome analysis and identified the prePIP1 gene from Solanum pimpinellifolium (SpprePIP1), as an ortholog of Arabidopsis prePIP1 encoding the precursor protein of PAMP-induced SSP 1. The expression level of SpprePIP1 is transcriptionally induced in tomato upon infection with Phytophthora infestans (P. infestans), the pathogen responsible for late blight. Overexpression of SpprePIP1 resulted in enhanced tomato resistance to P. infestans. In addition, exogenous application of SpPIP1, whether through spraying or irrigation, improved tomato resistance by enhancing the transcript accumulations of pathogenesis-related proteins, as well as reactive oxygen species and the jasmonic acid (JA) levels. Integrated analysis of transcriptomics and metabolomics revealed the potential contributions of JA and phenylpropanoid biosynthesis to SpPIP1-induced tomato immunity. Additionally, SpPIP1 may strengthen tomato resistance to salt stress through the ABA signaling pathway. Overall, our findings demonstrate that SpPIP1 positively regulates tomato tolerance to P. infestans and salt stress, making it a potential plant elicitor for crop protection in an environmentally friendly way.

Na+ transporter HKT1;2 reduces flower Na+ content and considerably mitigates the decline in tomato fruit yields under saline conditions

Abstract: We evaluated the salt tolerance of tomato cv Big Dena under both nongrafted conditions and when grafted on Maxifort rootstock, under a series of 5 salinity levels and 2 irrigation water composition types. The salinity levels of the irrigation water were -0.03, -0.15, -0.30, -0.45, and -0.60 MPa osmotic pressure (corresponding to specific electrical conductivity values of 1.2, 4.0, 8.5, 12, and 15.8 dS m-1, respectively). We salinized the irrigation water with either a mixture of salts with a predominant composition consisting of Na+-Ca2+-Cl- salts, a composition typical of coastal Mediterranean ground waters or, alternatively, a salt composition that was of mixed Na+-Ca2+-SO42--Cl- ions, a water composition more typical of interior continental basin ground waters such as those of the California Central Valley in the US. We determined that there were no statistically significant differences in tomato salt tolerance (fruit yield) relative to water type. This result indicates that in the range of Cl- concentrations tested in our experiment (up to 150 mmol L-1), Cl- is not an important factor in tomato yield reduction associated with salinity. The grafted Big Dena on Maxifort tomato plants exhibited increased yield both under control and elevated salinity levels relative to the nongrafted Big Dena plants. In contrast to absolute yield relationships, expression of salt tolerance in terms of relative yield, as salt tolerance is commonly expressed, provides the conclusion that grafted Big Dena on Maxifort tomato plants are slightly less salt tolerant than nongrafted Big Dena plants. Our data also indicate that, for tomato, decreased yield under saline conditions is well related to increased leaf Na+ concentrations.

Multiprotein bridging factor 1 (MBF1) is an ancient family of transcription coactivators that play a crucial role in the response of plants to abiotic stress. In this study, we analyzed the genomic data of five Solanaceae plants and identified a total of 21 MBF1 genes. The expansion of MBF1a and MBF1b subfamilies was attributed to whole-genome duplication (WGD), and the expansion of the MBF1c subfamily occurred through transposed duplication (TRD). Collinearity analysis within Solanaceae species revealed collinearity between members of the MBF1a and MBF1b subfamilies, whereas the MBF1c subfamily showed relative independence. The gene expression of SlER24 was induced by sodium chloride (NaCl), polyethylene glycol (PEG), ABA (abscisic acid), and ethrel treatments, with the highest expression observed under NaCl treatment. The overexpression of SlER24 significantly enhanced the salt tolerance of tomato, and the functional deficiency of SlER24 decreased the tolerance of tomato to salt stress. SlER24 enhanced antioxidant enzyme activity to reduce the accumulation of reactive oxygen species (ROS) and alleviated plasma membrane damage under salt stress. SlER24 upregulated the expression levels of salt stress-related genes to enhance salt tolerance in tomato. In conclusion, this study provides basic information for the study of the MBF1 family of Solanaceae under abiotic stress, as well as a reference for the study of other plants.

S-adenosylmethionine synthetase 1 confers drought and salt tolerance in transgenic tomato

The DEAD-box RNA helicases are involved in almost every aspect of RNA metabolism, associated with diverse cellular functions including plant growth and development, and their importance in response to biotic and abiotic stresses is only beginning to emerge. However, none of DEAD-box genes was well characterized in tomato so far. In this study, we reported on the identification and characterization of two putative DEAD-box RNA helicase genes, SlDEAD30 and SlDEAD31 from tomato, which were classified into stress-related DEAD-box proteins by phylogenetic analysis. Expression analysis indicated that SlDEAD30 was highly expressed in roots and mature leaves, while SlDEAD31 was constantly expressed in various tissues. Furthermore, the expression of both genes was induced mainly in roots under NaCl stress, and SlDEAD31 mRNA was also increased by heat, cold, and dehydration. In stress assays, transgenic tomato plants overexpressing SlDEAD31 exhibited dramatically enhanced salt tolerance and slightly improved drought resistance, which were simultaneously demonstrated by significantly enhanced expression of multiple biotic and abiotic stress-related genes, higher survival rate, relative water content (RWC) and chlorophyll content, and lower water loss rate and malondialdehyde (MDA) production compared to wild-type plants. Collectively, these results provide a preliminary characterization of SlDEAD30 and SlDEAD31 genes in tomato, and suggest that stress-responsive SlDEAD31 is essential for salt and drought tolerance and stress-related gene regulation in plants.

Beyond just being foot soldiers – osmotin like protein (OLP) and chitinase (Chi11) genes act as sentinels to confront salt, drought, and fungal stress tolerance in tomato

Changes of antioxidative enzymes and cell membrane osmosis in tomato colonized by arbuscular mycorrhizae under NaCl stress