title: Evaluation of a SUMO E2 Conjugating Enzyme Involved in Resistance to Clavibacter michiganensis Subsp. michiganensis in Solanum peruvianum, Through a Tomato Mottle Virus VIGS Assay.
abstract: Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker of tomato. Currently, no Solanum lycopersicum resistant varieties are commercially available, but some degree of Cmm resistance has been identified in Solanum peruvianum. Previous research showed up-regulation of a SUMO E2 conjugating enzyme (SCEI) transcript in S. peruvianum compared to S. lycopersicum following infection with Cmm. In order to test the role of SCEI in resistance to Cmm, a fragment of SCEI from S. peruvianum was cloned into a novel virus-induced gene-silencing (VIGS) vector based on the geminivirus, Tomato Mottle Virus (ToMoV). Using biolistic inoculation, the ToMoV-based VIGS vector was shown to be effective in S. peruvianum by silencing the magnesium chelatase gene, resulting in leaf bleaching. VIGS with the ToMoV_SCEI construct resulted in ~61% silencing of SCEI in leaves of S. peruvianum as determined by quantitative RT-PCR. The SCEI-silenced plants showed unilateral wilting (15 dpi) and subsequent death (20 dpi) of the entire plant after Cmm inoculation, whereas the empty vector-treated plants only showed wilting in the Cmm-inoculated leaf. The SCEI-silenced plants showed higher Cmm colonization and an average of 4.5 times more damaged tissue compared to the empty vector control plants. SCEI appears to play an important role in the innate immunity of S. peruvianum against Cmm, perhaps through the regulation of transcription factors, leading to expression of proteins involved in salicylic acid-dependent defense responses.
authors.affiliation: División Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A.C. San Luis Potosí, México.
authors.forename: Mayra J
authors.initials: MJ
authors.lastname: Esparza-Araiza
authors.affiliation: División Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A.C. San Luis Potosí, México.
authors.forename: Bernardo
authors.initials: B
authors.lastname: Bañuelos-Hernández
authors.affiliation: División Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A.C. San Luis Potosí, México.
authors.forename: Gerardo R
authors.initials: GR
authors.lastname: Argüello-Astorga
authors.affiliation: Facultad de Agronomía y Veterinaria, Universidad Autónoma de San Luis San Luis Potosí, México.
authors.forename: José P
authors.initials: JP
authors.lastname: Lara-Ávila
authors.affiliation: School of Environmental Sciences, University of Guelph Guelph, ON, Canada.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Goodwin
authors.affiliation: División Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A.C. San Luis Potosí, México.
authors.forename: María I
authors.initials: MI
authors.lastname: Isordia-Jasso
authors.affiliation: División Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A.C. San Luis Potosí, México.
authors.forename: Rosalba
authors.initials: R
authors.lastname: Castillo-Collazo
authors.affiliation: Laboratory of Agrogenomic Sciences, Universidad Nacional Autónoma de México, ENES-León León, México.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Rougon-Cardoso
authors.affiliation: División Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A.C. San Luis Potosí, México.
authors.forename: Ángel G
authors.initials: ÁG
authors.lastname: Alpuche-Solís
keywords: Clavibacter michiganensis subsp. michiganensis; SUMO-conjugating enzyme; Tomato Mottle Virus; bacterial canker; virus-induced gene-silencing