Record Details

Optimising parameters for biolistic gun-mediated genetic transformation of tea [Camellia sinensis (L.) O. Kuntze]

IR@CSIR-IHBT

View Archive Info
 
 
Field Value
 
Title Optimising parameters for biolistic gun-mediated genetic
transformation of tea [Camellia sinensis (L.) O. Kuntze]
 
Creator Saini, U
Kaur, D
Bhattacharya, Amita
Kumar, Sanjay
Singh, R D
Ahuja, Paramvir Singh
 
Subject Plant sciences
Plant Biotechnology
 
Description Genetic improvement of tea through breeding is difficult.Therefore, transgenic tea plants expressing the osmotin gene
from Nicotiana tabacum were produced using parameters optimised for biolistic-gun mediated transformation. During
optimisation, a total of 4,500 somatic embryos were bombarded using nine combinations of variable target distances
and burst pressures, while keeping the gap distance (0.6 cm) and macrocarrier flight distance (16 mm) constant. A total
of 90 independent, PCR-positive lines were generated. Southern hybridisation confirmed integration of the osmotin
gene in 26 out of 27 PCR-positive lines (three independent lines from each of the nine parameter combinations were
selected at random). Statistical analysis revealed that the efficiency of transgene integration was significantly affected
by target distance. Only those lines derived from somatic embryos bombarded with 1.0 μg plasmid DNA using a 7.58
MPa burst pressure and 9-cm target distance showed osmotin expression. This was evident from strong northern
hybridisation and RT-PCR signals. Leaves of 4-year-old transgenic plants growing in a contained polythene tunnel
showed improved osmotic adjustment in response to osmotic stress imposed by NaCl. The osmotic potentials of
transgenic leaves immersed in 100 mM or 200 mM NaCl solutions were more negative than those of non-transformed
control leaves.
 
Date 2012
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://ihbt.csircentral.net/1297/1/76_Optimising_parameters.pdf
Saini, U and Kaur, D and Bhattacharya, Amita and Kumar, Sanjay and Singh, R D and Ahuja, Paramvir Singh (2012) Optimising parameters for biolistic gun-mediated genetic transformation of tea [Camellia sinensis (L.) O. Kuntze]. Journal of Horticultural Science & Biotechnology, 87 (6). pp. 605-612.
 
Relation http://ihbt.csircentral.net/1297/