Volume 14, number 3
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Gene Stacking for Fungal Resistance in Plant Transformation Vector

Sonia Sharma1, Gurtej Singh1, Sadiq Pasha Shaik2, Gagandeep Singh3, Sumangala Bhat4 and Gaurav Sharma1

1Department of Natural Sciences, Sant Baba Bhag Singh University, Jalandhar 144030, India.

2Division of Biotechnology, Indian Institute of Horticulture Research, Hesaraghatta, Bengaluru 560089, India.

3Department of Agriculture, Mata Gujri College, Fatehgarh Sahib 140407, India.

4Department of Biotechnology, University of Agricultural Sciences, Dharwad-580 005, India.

Corresponding Author E-mail: g.sharma7346@gmail.com

DOI : http://dx.doi.org/10.13005/bbra/2563

ABSTRACT: Fungal diseases like early blight, late blight, fusarium wilt cause 30-40 per cent loss in fruit production. Form past decade many transgenic plants had been developed using genes encoding chitinases and glucanases with the objective of imparting fungal disease resistance. Since the genes encoding chitinase and glucanase act synergistically. The study was performed to construct plant transformation vector pRAGS carrying both ech42 and bgn under single T-DNA region. Initially, HindIII site at 5' end of earlier cloned bgn (T. harzianum) was removed using primers during reamplification of the gene. The amplicons were cloned into pTZ57R/T containing T overhangs at Eco321 site and transferred to E. coli DH5a and further to plant transformation vector pBI121 which was named as pRA121. In order to clone another gene (ech42) into pRA121, expression cassette from iHP vector was transferred to pRA121 and named as pRAG121. Further in order to gain XhoI site for cloning ech42 gene into pRAG121, ech42 (pSUM1) was cloned into pYES2/CT, named as pSAG1, ech42 from pSAG1 cloned with KpnI and XhoI in pRAG121 and named as pRAGS121. The vector constructed in the present study can be used to transform important crop plants to have enhanced resistance to fungal diseases.

KEYWORDS: Gene Stacking; Chitinase; Glucanase; Gene pyramiding; kb=Kilobase; bgn= beta glucanase; ech= Endochitinase; PCR= polymerase chain reaction

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Sharma S, Singh G, Shaik S. P, Singh G, Bhat S, Sharma G. Gene Stacking for Fungal Resistance in Plant Transformation Vector. Biosci Biotech Res Asia 2017;14(3).

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Sharma S, Singh G, Shaik S. P, Singh G, Bhat S, Sharma G. Gene Stacking for Fungal Resistance in Plant Transformation Vector. Biosci Biotech Res Asia 2017;14(3). Available from: https://www.biotech-asia.org/?p=27827

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