title: High efficiency transformation of banana [Musa acuminata L. cv. Matti (AA)] for enhanced tolerance to salt and drought stress through overexpression of a peanut salinity-induced pathogenesis-related class 10 protein.
abstract: Bananas and plantains (Musa spp. L.) are important subsistence crops and premium export commodity in several countries, and susceptible to a wide range of environmental and biotic stress conditions. Here, we report efficient, rapid, and reproducible Agrobacterium-mediated transformation and regeneration of an Indian niche cultivar of banana [M. acuminata cv. Matti (AA)]. Apical meristem-derived highly proliferative multiple shoot clump (MSC) explants were transformed with the Agrobacterium strain EHA105 harboring a binary vector pCAMBIA-1301 carrying hptII and uidA. Sequential agro-infiltration (10 min, 400 mmHg), infection (additional 35 min, Agrobacterium density A 600 = 0.8) and co-cultivation (18 h) regimen in 100 µM acetosyringone containing liquid medium were critical factors yielding high transformation efficiency (~81 %) corroborated by transient GUS expression assay. Stable transgenic events were recovered following two cycles of meristem initiation and selection on hygromycin containing medium. Histochemical GUS assay in several tissues of transgenic plants and molecular analyses confirmed stable integration and expression of transgene. The protocol described here allowed recovery of well-established putative transgenic plantlets in as little as 5 months. The transgenic banana plants could be readily acclimatized under greenhouse conditions, and were phenotypically similar to the wild-type untransformed control plants (WT). Transgenic plants overexpressing Salinity-Induced Pathogenesis-Related class 10 protein gene from Arachis hypogaea (AhSIPR10) in banana cv. Matti (AA) showed better photosynthetic efficiency and less membrane damage (P < 0.05) in the presence of NaCl and mannitol in comparison to WT plants suggesting the role of AhSIPR10 in better tolerance of salt stress and drought conditions.
authors.affiliation: Plant Developmental Biology and Transformation Lab, School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India.
authors.forename: Anjana
authors.initials: A
authors.lastname: Rustagi
authors.forename: Shalu
authors.initials: S
authors.lastname: Jain
authors.forename: Deepak
authors.initials: D
authors.lastname: Kumar
authors.forename: Shashi
authors.initials: S
authors.lastname: Shekhar
authors.forename: Mukesh
authors.initials: M
authors.lastname: Jain
authors.forename: Vishnu
authors.initials: V
authors.lastname: Bhat
authors.forename: Neera Bhalla
authors.initials: NB
authors.lastname: Sarin
mesh_terms: D000222:Adaptation, Physiological / Q000187:drug effects*; D015139:Blotting, Southern; D055864:Droughts; D004573:Electrolytes / Q000378:metabolism; D028521:Musa / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D018515:Plant Leaves / Q000187:drug effects / Q000254:growth & development; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D012038:Regeneration / Q000187:drug effects; D054712:Salinity; D012965:Sodium Chloride / Q000494:pharmacology*; D013312:Stress, Physiological / Q000187:drug effects*; D014170:Transformation, Genetic
chemical_list: D004573:Electrolytes; D010940:Plant Proteins; D012965:Sodium Chloride