Jalaja, N. and Maheshwari, P. and Naidu, K.R. and Kavi Kishor, P.B.
(2016)
In vitro regeneration and optimization of conditions for transformation methods in Pearl millet, Pennisetum glaucum (L.).
International Journal of Clinical and Biological Sciences, 1 (1).
pp. 34-52.
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Abstract
Pearl millet is a dual-purpose crop used for grain and fodder and is grown primarily in Asia and Africa,
where it occupies some 27 million ha. It is capable of growing on some of the poorest soils in dry, hot
regions of Africa and Asia, where, as a poor man’s source of dietary energy, it sustains a large proportion
of the populace. It is also grown in other countries where, under relatively more favorable conditions, it
provides grain for bullocks, dairy cows, and poultry. Downy mildew caused by Sclerospora graminicola
(Sacc.) J. Schroet. is the most widespread and destructive disease of pearl millet causing severe economic
losses. New genes can be introduced into this plant through Agrobacterium mediated and bombardment
genetic transformation for its genetic improvement, which is dependent on the availability of suitable in
vitro techniques. An efficient regeneration system has been developed for in vitro culture of pearl millet
(Pennisetum glaucum L.) from the immature inflorescence. High frequency callus and shoot regeneration
was obtained on Murashige and Skoog nutrient agar medium supplemented with 2mg/l 2,4-D and 0.2
mg/l NAA, 2 mg/l Kinetin and 30 g/l sucrose. On transfer to soil, the regenerated plantlets survived and
appeared to be morphologically similar to the normal seed-grown plants. Histological analysis revealed
the de novo origin of shoots from embryogenic callus in in vitro cultured pearl millet. Parameters
affecting transformation were optimized by assaying phosphinothricin resistance to transformed calli and
basta test for these leaves of plantlets after transferring to pots. These tissues appear to be susceptible to
Agrobacterium infection and Particle gun flow mediated transformation carrying pCAMBIA2300 with
osmotin and chitinase double construct and pPUR with bar genes, as well as shoot multiplication. The
embryogenic callus was found competent to take up the DNA, which was monitored by transient bar gene
with GV2600 at 0.6 O.D for Agrobacterium infection and 1μg/μl plasmid DNA from E.coli for cobombardment
was found to be compatible in giving transgenics.
| Item Type: |
Article
|
| Uncontrolled Keywords: |
co-bombardment transformation, phosphinothricin, basta, regeneration, pearl millet,
Pennisetum glaucum, immature inflorescence |
| Author Affiliation: |
Department of Biotechnology, Vignan’s University, Vadlamudi- 522213, India |
| Subjects: |
Crop Improvement |
| Divisions: |
Millet |
| Depositing User: |
Mr T L Gautham
|
| Date Deposited: |
10 Mar 2016 03:51 |
| Last Modified: |
21 Mar 2017 04:10 |
| URI: |
http://eprints.icrisat.ac.in/id/eprint/14218 |
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