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XSP10 and SlSAMT, Fusarium wilt disease responsive genes of tomato (Solanum lycopersicum L.) express tissue specifically and interact with each other at cytoplasm in vivo.

ABSTRACT: Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (Fol) is a major fungal disease of tomato (Solanum lycopersicum L.). Xylem sap protein 10 (XSP10) and Salicylic acid methyl transferase (SlSAMT) have been identified as putative negative regulatory genes associated with Fusarium wilt of tomato. Despite their importance as potential genes for developing Fusarium wilt disease tolerance, very little knowledge is available about their expression, cell biology, and functional genomics. Semi-quantitative and quantitative real-time PCR expression analysis of XSP10 and SlSAMT, in this study, revealed higher expression in root and flower tissue respectively in different tomato cultivars viz. Micro-Tom (MT), Arka Vikas (AV), and Arka Abhed (AA). Therefore, the highly up-regulated expression of XSP10 and SlSAMT in biotic stress susceptible tomato cultivar (AV) than a multiple disease resistant cultivar (AA) suggested the disease susceptibility nature of these genes for Fusarium wilt. Sub-cellular localization analysis through the expression of gateway cloning constructs in tomato protoplasts and seedlings showed the predominant localization of XSP10 in the nucleus and SlSAMT at the cytoplasm. A strong in vivo protein-protein interaction of XSP10 with SlSAMT at cytoplasm from bi-molecular fluorescent complementation study suggested that these two proteins function together in regulating responses to Fusarium wilt tolerance in tomato.

Supplementary information

The online version contains supplementary material available at 10.1007/s12298-021-01025-y.

SUBMITTER: Debbarma J

PROVIDER: S-EPMC8295444 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature

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XSP10 and SlSAMT, Fusarium wilt disease responsive genes of tomato (Solanum lycopersicum L.) express tissue specifically and interact with each other at cytoplasm in vivo.

Debbarma Johni J Saikia Banashree B Singha Dhanawantari L DL Maharana Jitendra J Velmuruagan Natarajan N Dekaboruah Hariprasanna H Arunkumar Kallare P KP Chikkaputtaiah Channakeshavaiah C

Physiology and molecular biology of plants : an international journal of functional plant biology 20210628 7

Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (Fol) is a major fungal disease of tomato (Solanum lycopersicum L.). Xylem sap protein 10 (XSP10) and Salicylic acid methyl transferase (SlSAMT) have been identified as putative negative regulatory genes associated with Fusarium wilt of tomato. Despite their importance as potential genes for developing Fusarium wilt disease tolerance, very little knowledge is available about their expre ...[more]

PMID: 34366597

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Project description:The antagonistic efficacy of a biocontrol agent in combination with oilseed cake against fungal phytopathogens has been sparsely explored. The present study aimed to evaluate the antifungal activity of a biocontrol agent (Trichoderma harzianum MTCC 3928) formulated with oilseed cake (OSC) against Fusarium oxysporum f. sp. lycopersici responsible for causing vascular wilt in Solanum lycopersicum. In in vitro studies, OSC of mustard (Brassica juncea) exhibited significant mycelial inhibition against the pathogen. The volatile plate assay showed mycelial inhibition of 70 and 40% with unautoclaved and autoclaved mustard cakes, respectively. The aqueous extract (10% v/v) of the mustard cake was the most effective with 47.3% mycelial inhibition of pathogen over control. In addition, volatiles and aqueous extract of mustard cake subjected to GC-MS analysis revealed a range of antifungal bioactive compounds with hexanedioic acid, dioctyl ester (16.57%), and oleic acid trimethylsilyl ester (12.41%) being predominant compounds. In in vitro studies, it was noticed that the T. harzianum strain was compatible with mustard cake, and hence used as a growth substrate for its mass multiplication. SEM analysis revealed no distortion in spores and mycelium of T. harzianum grown on the mustard cake. Further, seed germination assay suggested the optimum concentration of mustard cake (10%) supporting the germination rate and economics of formulation development. In in planta assay, the combination of biocontrol agent and mustard cake showed 48% disease reduction, and ~ 40% with T. harzianum alone in comparison to untreated control. Also, the combination of mustard cake and T. harzianum significantly enhanced the growth parameters of S. lycopersicum. The findings of the current study identified an environmentally friendly alternative for mitigation of Fusarium wilt, thereby providing a sustainable option for mitigation of wilt disease and enhancement of plant health.

Project description:Fusarium wilt is a major devastating fungal disease of tomato (Solanum lycopersicum L.) caused by Fusarium oxysporum f. sp. lycopersici (Fol) which reduces the yield and production. Xylem sap protein 10 (XSP10) and Salicylic acid methyl transferase (SlSAMT) are two putative negative regulatory genes associated with Fusarium wilt of tomato. Fusarium wilt tolerance in tomato can be developed by targeting these susceptible (S) genes. Due to its efficiency, high target specificity, and versatility, CRISPR/Cas9 has emerged as one of the most promising techniques for knocking out disease susceptibility genes in a variety of model and agricultural plants to increase tolerance/resistance to various plant diseases in recent years. Though alternative methods, like RNAi, have been attempted to knock down these two S genes in order to confer resistance in tomato against Fusarium wilt, there has been no report of employing the CRISPR/Cas9 system for this specific intent. In this study, we provide a comprehensive downstream analysis of the two S genes via CRISPR/Cas9-mediated editing of single (XSP10 and SlSAMT individually) and dual-gene (XSP10 and SlSAMT simultaneously). Prior to directly advancing on to the generation of stable lines, the editing efficacy of the sgRNA-Cas9 complex was first validated using single cell (protoplast) transformation. In the transient leaf disc assay, the dual-gene editing showed strong phenotypic tolerance to Fusarium wilt disease with INDEL mutations than single-gene editing. In stable genetic transformation of tomato at the GE1 generation, dual-gene CRISPR transformants of XSP10 and SlSAMT primarily exhibited INDEL mutations than single-gene-edited lines. The dual-gene CRISPR-edited lines (CRELs) of XSP10 and SlSAMT at GE1 generation conferred a strong phenotypic tolerance to Fusarium wilt disease compared to single-gene-edited lines. Taken together, the reverse genetic studies in transient and stable lines of tomato revealed that, XSP10 and SlSAMT function together as negative regulators in conferring genetic tolerance to Fusarium wilt disease.

Dataset Information

XSP10 and SlSAMT, Fusarium wilt disease responsive genes of tomato (Solanum lycopersicum L.) express tissue specifically and interact with each other at cytoplasm in vivo.

Supplementary information

Publications

<i>XSP10</i> and <i>SlSAMT</i>, Fusarium wilt disease responsive genes of tomato (<i>Solanum lycopersicum</i> L.) express tissue specifically and interact with each other at cytoplasm in vivo.

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