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. 2009 Feb 26;14(4):307–313. doi: 10.1007/s12298-008-0028-9

Tissue specific response of Agrobacterium tumefaciens attachment to Sorghum bicolor (L) Moench

Anju Verma 1, Vikrant Nain 1, Chetana Kumari 1, Santosh Kumar Singh 1, M Lakshmi Narasu 2, P Ananda Kumar 1,✉
PMCID: PMC3550632  PMID: 23572896

Abstract

Agrobacterium mediated genetic transformation of plants have advantages over other methods, especially for making single copy transgenic plants with reduced chances of gene silencing and instability. However, monocotyledonous plant species could not utilize the full potential of this system because of possible limitations in Agrobacterium interaction with monocot plant cells. Agrobacterium attachment as a factor in genetic transformation was studied in the leaf, shoot apex, and leaf derived callus of sorghum (Sorghum bicolor (L) Moench). Pre-induction of Agrobacterium with acetosyringone was found necessary for Agrobacterium attachment to sorghum tissues. All the explants responded positively, with preferential Agrobacterium attachment and colonization around the tissues having actively dividing cells. Callus proved to be the best explant for Agrobacterium attachment as observed in scanning electron microscopy and transient GUS expression. Loss of Agrobacterium attachment was observed with an increase in the degree of tissue differentiation.

Key words: Genetic transformation, Acetosyringone, Scanning electron microscopy, Transient gene expression, GUS assays, qRT-PCR

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Abbreviations

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid

BAP

Benzyl amino purine

ADS

Adenine sulphate

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