TRANSFORMATION OF ZEA MAYS USING DREB1A GENE FOR ABIOTIC STRESSES TOLERANCE

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SHAMS M.R.; RAHNAMA H.; BIHAMTA M.R.; KHODARAHMI M.

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Paper Information

Journal: IRANIAN JOURNAL OF AGRONOMY AND PLANT BREEDING Year:2013 | Volume:9 | Issue:1 Page(s): 75-89

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Title

TRANSFORMATION OF ZEA MAYS USING DREB1A GENE FOR ABIOTIC STRESSES TOLERANCE

Author(s)

SHAMS M.R. | RAHNAMA H. | BIHAMTA M.R. | KHODARAHMI M. | Issue Writer Certificate

Keywords

ABIOTIC STRESSESQ2

DREB1A GENEQ2

GENE GUNQ3

GENE TRANSFORMATIONQ2

IMMATURE EMBRYOSQ2

ZEA MAYSQ2

Abstract

In order to genetic transformation of ZEA MAYS using DREB1A GENE for tolerance to ABIOTIC STRESSES, recombinant Binary vector (pCRDREB) was constructed at first stage. To produce a binary vector containing the DREB1A GENE, the 651 bp fragment which had been cloned in pTZ57R vector, was digested with EcoRI and then subcloned into the homologous site of the pUC18 cloning vector. Subsequently, The 910 bp fragment of rd29A promoter from Arabidopsis and the 260 bp fragment of NOS terminator from pBI121 binary vector were amplified by PCR and specific primers Then PCR products of rd29A and nos were digested with HindIII, XbaI and also EcoRI and PstI, respectively. Then, they were cloned into the pUC18-DREB cloning vector. Rd-DREB-nos Gene-cassette were digested with EcoRI and HindIII, prior to subcloning into the homologous sites of the pCAMBIA3300 binary vector. ZEA MAYS (A188, S61 and B73 var) were used in this study and they were planted in vases. Ears were harvested 10–14 days after pollination when they reached 1-2mm long. After they sterilized, immature zygotic embryos from the harvested cobs were cultured on pre-culture medium (N6P) for four days. Embryos were subjected to osmoticum treatment (N6Y) 3h before bombardment. DREB1A GENE was bombarded to the target tissue using the biolistic gun. Secondary embryos were transferred on N6 Rest medium for a period of seven days and afterwards they were delivered to selective media containing PPT. Resistant call cultured on the regenerating and germinating media and plantlets removed to rooting medium. Finally genomic DNA was extracted from maize transformants. The presence of the bar (Herbicide resistant) and DREB1A (resistance to ABIOTIC STRESSES) genes was confirmed by PCR analysis.

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APA: Copy

SHAMS, M.R., RAHNAMA, H., BIHAMTA, M.R., & KHODARAHMI, M.. (2013). TRANSFORMATION OF ZEA MAYS USING DREB1A GENE FOR ABIOTIC STRESSES TOLERANCE. IRANIAN JOURNAL OF AGRONOMY AND PLANT BREEDING, 9(1), 75-89. SID. https://sid.ir/paper/190334/en

Vancouver: Copy

SHAMS M.R., RAHNAMA H., BIHAMTA M.R., KHODARAHMI M.. TRANSFORMATION OF ZEA MAYS USING DREB1A GENE FOR ABIOTIC STRESSES TOLERANCE. IRANIAN JOURNAL OF AGRONOMY AND PLANT BREEDING[Internet]. 2013;9(1):75-89. Available from: https://sid.ir/paper/190334/en

IEEE: Copy

M.R. SHAMS, H. RAHNAMA, M.R. BIHAMTA, and M. KHODARAHMI, “TRANSFORMATION OF ZEA MAYS USING DREB1A GENE FOR ABIOTIC STRESSES TOLERANCE,” IRANIAN JOURNAL OF AGRONOMY AND PLANT BREEDING, vol. 9, no. 1, pp. 75–89, 2013, [Online]. Available: https://sid.ir/paper/190334/en

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