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Overexpression of the ascorbate peroxidase through enhancer‐trapped pSB111 bar vector for alleviating drought stress in rice
Author(s) -
Gorripati Srinivas,
Konka Rajasekhar,
Panditi Shravana Kumar,
Velagapudi Kavitha,
Jeevigunta Naveena Lavanya Latha
Publication year - 2021
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.202000725
Subject(s) - agrobacterium tumefaciens , biology , gene , expression vector , plasmid , agrobacterium , enhancer , transformation (genetics) , reporter gene , insert (composites) , microbiology and biotechnology , gus reporter system , oryza sativa , genetics , green fluorescent protein , gene expression , recombinant dna , mechanical engineering , engineering
Rice ( Oryza sativa L.) plant growth and productivity is adversely affected by various stress factors. Overexpression of drought tolerance‐related genes is one of the best approaches for developing drought‐resistant transgenics. Agrobacterium tumefaciens has been widely used in generating transgenic plants through plasmid vector to obtain desired characteristics and to know the specific expression profiles of genes in the plant. The enhancer trap method was developed to know the specific expression of genes at different stages of growth by entrapping the genes of an organism. In the present study, we designed a vector molecule with a feature of promoting the expression of a specific gene more than four times than its normal expression and it is useful for efficient transformation to higher plants by utilizing the trans configuration of vir genes of the plasmid A. tumefaciens , to transfer right and left sequence bordered of transferred DNA (T‐DNA) into the nuclear genome of plants. We developed a binary vector consisting of 1.8‐kb green fluorescent protein (GFP) cassette as a reporter gene and 1.4‐kb tetramer of CaMv35S enhancer (4XEn) were cloned at Hin dIII site of pSB11 bar intermediate vector to tag and know the genes and their expression profiles, then mobilized into A. tumefaciens to produce a super‐binary vector pSB111‐bar‐4XEn‐GFP. The resultant construct was confirmed by polymerase chain reaction and restriction digestion methods. Finally, we discuss the role of overexpressed ascorbate peroxidase in drought stress.