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A method to overcome the waxy surface, cell wall thickening and polyphenol induced necrosis at wound sites - the major deterrents to Agrobacterium mediated transformation of bamboo, a woody monocot

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Title A method to overcome the waxy surface, cell wall thickening
and polyphenol induced necrosis at wound sites - the major
deterrents to Agrobacterium mediated transformation
of bamboo, a woody monocot
 
Creator Sood, Priyanka
Bhattacharya, Amita
Joshi, Robin
Gulati, Ashu
Chanda , Sanjoy
Sood, A
 
Subject Plant sciences
Plant Biotechnology
 
Description The method is the first successful report of Agrobacterium
mediated genetic transformation of the commercially
important bamboo, Dendrocalamus hamiltonii. It
shows how the resistance provided by the somatic embryos
of this woody monocot can be overcome using a simple and
effective method. The method thus standardized can be also
used for the genetic transformation of other important
bamboos. Identification of the factors responsible for the
resistance of the somatic embryos to Agrobacterium infection
was an absolute requirement for devising a successful
method. Necrosis due to polyphenol oxidation, lack of differentiation
due to cell wall thickening at wound sites, waxy
surfaces of somatic embryos with anti-microbial properties
were found to prevent Agrobacterium attachment and infection.
Therefore, the somatic embryos were transformed with
fresh overnight grown Agrobacterium culture containing
500 mg/l polyvinylpyrrolidone (PVP) and 0.01 % Tween-
20 as surfactant followed by co-cultivation on Murashige
and Skoog (MS) medium containing the vir gene inducer
acetosyringone (100 μM) and 1 mg/l 6-Benzylaminopurine
BAP for 2 days. Persistent GUS expression and strong
positive signals in PCR, slot blot and Southern hybridization
confirmed successful genetic transformation.
 
Date 2013
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/1354/1/111_A_method.pdf
Sood, Priyanka and Bhattacharya, Amita and Joshi, Robin and Gulati, Ashu and Chanda , Sanjoy and Sood, A (2013) A method to overcome the waxy surface, cell wall thickening and polyphenol induced necrosis at wound sites - the major deterrents to Agrobacterium mediated transformation of bamboo, a woody monocot. Journal of Plant Biochemistry and Biotechnology.
 
Relation http://ihbt.csircentral.net/1354/