LARGE SCALE PRODUCTION OF Cry 1Aa PLANTS IN SORGHUM
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Date
2005
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ACHARYA N. G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD
Abstract
Sorghum is an important dryland crop used as food, feed and fodder. The
increasing shift in the demand for sorghum from staple food to industrially
preferred commodity in poultry, bakery and brewing industries requires
large scale production according to the needs of different sectors.
Therefore, it is important to develop and deploy novel technologies such as
genetic transformation to increase the grain quality and resistance to pests
that are difficult to breed by conventional means. Two principal methods
viz., Agrobacterium infection and particle bombardment were used for
genetic transformation. In vitro response of sorghum was low,
monocalcitrant and was highly genotype dependent. Tissue culture and
regeneration response was low in sorghum when compared to other cereals.
Agrobacterium method of transformation was preferred because of its cost
efficacy and low copy number. But regeneration of explants was very low
due to overgrowth of Agrobacterium. Hence during the course of study
alternate methods of Agrobacterium infection were studied using two
inbred lines AKMS 14B and 296B. Different media adjuvants were studied
to control Agrobacterium growth. AgNO3 and CuSO4 were effective in
controlling the bacterial growth. Gus expression was studied in
cotransformation experiments using two individual genes viz., pDM805
and pCAMBIA 3300 cry 1Aa. Gus results revealed that they were
independently transformed without influencing each other. A non tissue
culture approach was done to produce transformants at large scale. Though
transient gus expression was present in the immature embryos, mature
seeds were not selected in basta screening. Putative transformants were
evaluated through basta spray tests and PCR analysis using bar and Cry 1B
primers. Results of basta swab test were in agreement with PCR results.
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Keywords
LARGE, SCALE, PRODUCTION, Cry, 1Aa, PLANTS, SORGHUM