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

被引:30
|
作者
Rustagi, Anjana [1 ]
Jain, Shalu [1 ]
Kumar, Deepak [1 ]
Shekhar, Shashi [1 ]
Jain, Mukesh [1 ]
Bhat, Vishnu
Sarin, Neera Bhalla [1 ]
机构
[1] Jawaharlal Nehru Univ, Sch Life Sci, Plant Dev Biol & Transformat Lab, New Delhi 110067, India
关键词
Abiotic stress; Agrobacterium; Arachis hypogaea; Multiple shoot clumps; Musa spp; PR proteins; Transgenic; AGROBACTERIUM-MEDIATED TRANSFORMATION; EMBRYOGENIC-CELL SUSPENSIONS; EXPRESSION; PLANTS; ESTABLISHMENT; CULTURES; GENE; AAA;
D O I
10.1007/s12033-014-9798-1
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 A mu M acetosyringone containing liquid medium were critical factors yielding high transformation efficiency (similar to 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.
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页码:27 / 35
页数:9
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  • [1] 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
    Anjana Rustagi
    Shalu Jain
    Deepak Kumar
    Shashi Shekhar
    Mukesh Jain
    Vishnu Bhat
    Neera Bhalla Sarin
    Molecular Biotechnology, 2015, 57 : 27 - 35
  • [2] Ectopic overexpression of a salt stress-induced pathogenesis-related class 10 protein (PR10) gene from peanut (Arachis hypogaea L.) affords broad spectrum abiotic stress tolerance in transgenic tobacco
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    Kumar, Deepak
    Jain, Mukesh
    Chaudhary, Prerna
    Deswal, Renu
    Sarin, Neera Bhalla
    PLANT CELL TISSUE AND ORGAN CULTURE, 2012, 109 (01) : 19 - 31
  • [3] Ectopic overexpression of a salt stress-induced pathogenesis-related class 10protein (PR10) gene from peanut (Arachis hypogaea L.) affords broad spectrum abiotic stress tolerance in transgenic tobacco
    Shalu Jain
    Deepak Kumar
    Mukesh Jain
    Prerna Chaudhary
    Renu Deswal
    Neera Bhalla Sarin
    Plant Cell, Tissue and Organ Culture (PCTOC), 2012, 109 : 19 - 31