<i>Ralstonia solanacearum</i> virulence in tomato seedlings inoculated by leaf clipping

Kumar, Rahul; Barman, Anjan; Phukan, Tarinee; Kabyashree, Kristi; Singh, Niraj; Jha, Gopaljee; Sonti, Ramesh V.; Genin, Stéphane; Ray, Suvendra Kumar

doi:10.1111/ppa.12628

Cited by 33 publications

(36 citation statements)

References 37 publications

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“…Colonization of F1C1 in the inoculated eggplant seedlings was demonstrated by the GUS staining of the infected seedlings as performed previously in tomato seedlings (Kumar et al 2017). GUS staining was observed along the shoot region of the seedlings suggesting that growth and migration of the bacteria occurred towards the root region from the inoculated site in the cotyledon leaves along with the shoot (Fig.…”

Section: Resultsmentioning

confidence: 56%

“…The other possible reason may be that the pathogenicity assays used till date are more consistent and reproducible in tomato than in eggplant. In this present study we have demonstrated that the leaf-clip inoculation method is an easy and stable method for evaluating the virulence of R. solancearum in eggplant seedlings, which was ever successfully used to test R. solanacearum virulence in tomato seedlings (Kumar et al 2017). This is an easy and stable method of inoculation in which the juvenile seedlings are maintained in 1.5 or 2.0 mL microfuge tubes.…”

Section: Introductionmentioning

confidence: 74%

“…A consistent 100% death of the inoculated eggplant seedlings was recorded during each infection set-up. It was interesting for us because, in the case of tomato seedlings, some of the inoculated seedlings (~10%) did not exhibit the disease symptom (Kumar et al 2017). This indicated that the eggplant seedlings may be more susceptible to R. solanacearum infection than the tomato seedlings.…”

Section: Resultsmentioning

confidence: 99%

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Ralstonia solanacearum virulence in eggplant seedlings by the leaf-clip inoculation

et al. 2019

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Ralstonia solanacearum causes a lethal bacterial wilt disease in numerous plants including important vegetable crops such as eggplant and tomato. One of the difficulties in studying virulence of this bacterium in different host plants is the development of an easy and stable pathogenicity assay. Recently we described a leaf-clip inoculation method to study its pathogenicity at the cotyledon stage of tomato seedlings. Hereafter, we demonstrated the leafclip inoculation method to be equally efficient for studying R. solanacearum pathogenicity in the cotyledon stage of eggplant seedlings. Our study revealed eggplant seedlings to be highly susceptible to R. solanacearum as compared to tomato seedlings, illustrated by appearance of disease symptoms in significantly higher number of seedlings. We also tested the virulence of several global transcription regulator mutants of R. solanacearum including hrpB, hrpG and phcA in eggplant seedlings. The phcA mutant was found to be only moderately virulence deficient in eggplant seedlings but was significantly reduced in virulence in tomato. This is indicative of some host specific responses towards certain pathogenicity functions of R. solanacearum, which are markedly different in tomato and eggplant seedlings. Apart from being economical in requiring less labor, time and space, this simple gnotobiotic leaf-clip inoculation method is anticipated to be helpful in further exploring the interaction between R. solanacearum and eggplant seedlings at the cotyledon stage.

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Section: Resultsmentioning

confidence: 56%

Section: Introductionmentioning

confidence: 74%

Section: Resultsmentioning

confidence: 99%

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Ralstonia solanacearum virulence in eggplant seedlings by the leaf-clip inoculation

et al. 2019

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“…Bacterial inoculums were used for infection by root inoculation approach in 6-7 days old tomato seedlings as stated above. After third day of post inoculation, tomato seedlings were employed for colonization study and infected seedlings were surface sterilized by the method of Kumar et al ., (2016) (Kumar et al , 2017). After surface sterilization, seedlings were observed under the fluorescence microscopy (EVOS FL, Life technologies) for bacterial colonization study in 40X magnification adjusted in RFP filter.…”

Section: Methodsmentioning

confidence: 99%

A Rapid Screening of Resistant Tomato Cultivars in Early Seedling Stage AgainstRalstonia solanacearumBacterial Wilt Under Soil and Nutrients Free Conditions

Singh

2020

Preprint

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This work reports a rapid screening study of tomato (Solanum lycopersicum) cultivars resistant to R. solanacearum (F1C1) bacterial wilt in early seedling stage ( 6-7 days old, cotyledon leaf stage) under soil and nutrients free condition. This study was performed in 1.5-2.0 ml microfuge tube where root inoculation approach has been used during infection under gnotobiotic condition and got completed within 2 weeks. After continuous post inoculation observation of disease progression in all nine recruited tomato cultivar, Ayush was found to be the most resistant (~15% wilting) while the Ruby tomato cultivar was found to be the most susceptible (~95% wilting) against R. solanacearum bacterial wilt. This comparative wilting and survivability study among nine different tomato cultivar under same conditions indicated that early seedling stage of tomato may be a potent and helpful tool for an easy and rapid screening of resistant tomato cultivar against R. solanacearum bacterial wilt disease at large scale level in a very economical way in terms of time, space, labor and cost. Over and above The soil and nutrients free approach presented here may be helpful for the rapid selection and development of resistant tomato cultivar and new plant protection strategies against R. solanacearum bacterial wilt disease. W

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“…To demonstrate the functionality of GG-assembled expression modules, we first focused on the construction of an expression cassette containing the uidA gene. The uidA reporter gene was chosen as an Rssc strain expressing this reporter is easily traceable in infected tomato plants (Kumar et al, 2016). The expression construct was generated by fusing an eps promoter (LI A-B), the uidA CDS (LI C-D), the GFP CDS (LI D-E), a Tfd terminator (LI E-F) and a tetracycline resistance gene (LI F-G) via BsaI cut-ligation into an LII destination vector (Fig.…”

Section: Gg-based Assembly Of a Uida-gfp Expression Cassette For Use mentioning

confidence: 99%

A modular toolbox for Golden‐Gate‐based plasmid assembly streamlines the generation of Ralstonia solanacearum species complex knockout strains and multi‐cassette complementation constructs

Schandry

Lahaye

2018

Molecular Plant Pathology

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Members of the Ralstonia solanacearum species complex (Rssc) cause bacterial wilt, a devastating plant disease that affects numerous economically important crops. Like other bacterial pests, Rssc injects a cocktail of effector proteins via the bacterial type III secretion system into host cells that collectively promote disease. Given their functional relevance in disease, the identification of Rssc effectors and the investigation of their in planta function are likely to provide clues on how to generate pest-resistant crop plants. Accordingly, molecular analysis of effector function is a focus of Rssc research. The elucidation of effector function requires corresponding gene knockout strains or strains that express the desired effector variants. The cloning of DNA constructs that facilitate the generation of such strains has hindered the investigation of Rssc effectors. To overcome these limitations, we have designed, generated and functionally validated a toolkit consisting of DNA modules that can be assembled via Golden-Gate (GG) cloning into either desired gene knockout constructs or multi-cassette expression constructs. The Ralstonia-GG-kit is compatible with a previously established toolkit that facilitates the generation of DNA constructs for in planta expression. Accordingly, cloned modules, encoding effectors of interest, can be transferred to vectors for expression in Rssc strains and plant cells. As many effector genes have been cloned in the past as GATEWAY entry vectors, we have also established a conversion vector that allows the implementation of GATEWAY entry vectors into the Ralstonia-GG-kit. In summary, the Ralstonia-GG-kit provides a valuable tool for the genetic investigation of genes encoding effectors and other Rssc genes.

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Ralstonia solanacearum virulence in tomato seedlings inoculated by leaf clipping

Cited by 33 publications

References 37 publications

Ralstonia solanacearum virulence in eggplant seedlings by the leaf-clip inoculation

Ralstonia solanacearum virulence in eggplant seedlings by the leaf-clip inoculation

A Rapid Screening of Resistant Tomato Cultivars in Early Seedling Stage AgainstRalstonia solanacearumBacterial Wilt Under Soil and Nutrients Free Conditions

A modular toolbox for Golden‐Gate‐based plasmid assembly streamlines the generation of Ralstonia solanacearum species complex knockout strains and multi‐cassette complementation constructs

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