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Full Length Research Paper
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Effect of various amino
acids on shoot regeneration of sugarcane (Sacchrum
officinarum L.)
Shaheen Asad*, Muhammad Arshad*, Shahid Mansoor and Yusuf
Zafar
Plant
Biotechnology Division, National Institute for Biotechnology
and Genetic Engineering [NIBGE], P.O.Box-577, Jhang Road,
Faisalabad, Pakistan.
*Corresponding author: E-mail:
arshadchbt@yahoo.com
or sasad@nibge.org.
Abbreviations:
Gln, Glutamine; Asn, asparagines; Cys,
cysteine; Arg, arginine; Gly, glycin; CW,
coconut water; ca, approximately; ctr,
control; 2,4-D, dichlorophenoxyacetic acid; GRPs,
glycine-rich proteins.
Accepted
26 October, 2007 |
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Abstract |
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An efficient regeneration protocol from sugarcane callus has
been developed through inclusion of amino acids in
regeneration medium. Sugarcane (Saccharum officinarum
L.) SP-241 callus was induced from meristematic explants
cultured on Murashige and Skoog medium supplemented with B5
vitamins containing 13.6 µM 2-4, dichlorophenoxyacetic acid,
0.05% (w/v) casein hydrolysate, 10% (v/v) coconut water and
3% glucose. Five levels (0.1, 0.25, 0.5, 0.75, 1.0 mM) of
five different amino acids (glutamine, asparagine, glycine,
cysteine and arginine) were
tested on the same medium containing 6.8 µM 2, 4-D to
compare their ability to induce somatic embryogenesis and
shoot regeneration from six week old callus. Among the
tested amino acids, glycine (0.75 mM), arginine (0.5 mM) and
cysteine (0.25 mM) showed significant effect on smatic
embryogeensis (94%) and shoot production as compared to
non-amino acid medium. Of the evaluated amino acids, glycine
was most effective to promote somatic embryogeensis and
maximum shoot regeneration. Except low callus growth index (G.I)
amino acid treatment resulted in high percentages of somatic
embryogenesis and shoot regeneration as compared to
non-amino acid medium. Regenerated shoots when transferred
to same medium supplemented with 19.7 µM IBA, grew normal
and developed roots.
These results indicate the efficacy of amino acids in
stimulating sugarcane plant regeneration from non
embryogenic callus, and may be suitable for future use in
genetic transformation studies to enhance regeneration of
transgenic sugarcane plants.
Key
words:
Saccharum officinarum L, amino acids, embryogenic
calli, plant regeneration, 2, 4 dichlorophenoxyacetic acid. |
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