Articles
BIOLISTIC-MEDIATED GENETIC TRANSFORMATION OF PRICKLY-PEAR CACTUS (OPUNTIA FICUS-INDICA MILL.)
Article number
811_32
Pages
255 – 258
Language
English
Abstract
Opuntia ficus-indica is a cactus species important in agricultural economies throughout arid and semiarid parts of the world.
It is an important source of vegetable and forage in areas with adverse environmental conditions.
Efforts in developing transformation systems for this species are limited.
Nevertheless, in the past we have obtained promising results when biolistic were applied on both callus cultures and shoot apical regions.
However, no molecular evidence for transgenic plants was presented.
Recently, an Agrobacterium-mediated system was developed.
The aim of the present work was to develop a transformation system for cactus pear using the biolistic process.
Apical regions were bombarded with the plasmid pAG1, which contains the gus gene and the Atahas gene, that confers resistance to imazapyr, a herbicidal molecule that concentrates in the apical meristematic region of the plant.
Transient and stable GUS activities were monitored on herbicide tolerant regenerated shoots and plantlets.
Out of 120 bombarded explants, four survived in 300 nM imazapyr and one showed gus gene expression.
Genetic transformation was demonstrated by PCR. The transformation frequency obtained by the system reported here was of 0.8%. This method may be useful for routine transformation and introduction of several important genes in prickly pear cactus.
It is an important source of vegetable and forage in areas with adverse environmental conditions.
Efforts in developing transformation systems for this species are limited.
Nevertheless, in the past we have obtained promising results when biolistic were applied on both callus cultures and shoot apical regions.
However, no molecular evidence for transgenic plants was presented.
Recently, an Agrobacterium-mediated system was developed.
The aim of the present work was to develop a transformation system for cactus pear using the biolistic process.
Apical regions were bombarded with the plasmid pAG1, which contains the gus gene and the Atahas gene, that confers resistance to imazapyr, a herbicidal molecule that concentrates in the apical meristematic region of the plant.
Transient and stable GUS activities were monitored on herbicide tolerant regenerated shoots and plantlets.
Out of 120 bombarded explants, four survived in 300 nM imazapyr and one showed gus gene expression.
Genetic transformation was demonstrated by PCR. The transformation frequency obtained by the system reported here was of 0.8%. This method may be useful for routine transformation and introduction of several important genes in prickly pear cactus.
Authors
A.R.R. Cruz, E.L. Soares, F.A.P. Campos, F.J.L. Aragão
Keywords
biolistic, gene expression, genetic transformation, Opuntia
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