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Please use this identifier to cite or link to this item:
http://krishi.icar.gov.in/jspui/handle/123456789/9162
Title: | In Vitro Plant Regeneration and Genetic Transformation of Dichanthium annulatum |
Other Titles: | Not Available |
Authors: | J Kumar, S M. Shukla, V Bhat, S Gupta, M.G. Gupta |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR::Directorate of Soybean Research |
Published/ Complete Date: | 2005-08-12 |
Project Code: | Not Available |
Keywords: | in-vitro regeneration, Transformation, Dichanthium |
Publisher: | Not Available |
Citation: | Not Available |
Series/Report no.: | Not Available; |
Abstract/Description: | Optimization of in vitro plant regeneration and genetic transformation of apomictic species such as Dichanthium annulatum would enable transfer of desirable genes. Seven genotypes of this grass species were screened through mature seed explant for embryogenic callus induction, callus growth and quality (color and texture), and shoot induction. Genotype IG-1999, which produced highly embryogenic, rapidly growing good-quality callus capable of regenerating at a high frequency, was selected for transformation experiments. Using a binary vector (pCAMBIA1305), frequency of GUS expression was compared between two methods of transformation. Bombardment of embryogenic calli with gold particles coated with pCAMBIA1305 at a distance of 11 cm, pressure of 4 bars, and vacuum of 27 Hg passing through 100 μM mesh produced maximum GUS expression (23%). Agrobacterium infection was maximum at an optical density of 2.0 when cocultured under vacuum for 15 min and cocultivated for 3 days at 28°C in constant dark on MS medium of pH 5.8 with 3 mg/l 2,4-D, and 400 μM acetosyringone. Among two binary vectors used for Agrobacterium-mediated transformation, pCAMBIA1301 showed higher frequency of GUS expression while pCAMBIA1305 recorded more of the GUS spots per callus. Supplementation of acetosyringone in the cocultivation medium was found indispensable for Agrobacterium-mediated transformation. Injuring the calli through gold particle bombardment before their cocultivation with Agrobacterium improved the transformation efficiency. Several transgenic plants were developed using the PIG method, while stable GUS-expressing calli were multiplied during selection on MS medium containing 250 mg/l cefotaxime and 50 mg/l hygromycin, incubated in constant dark. A highly significant difference was observed between two methods of transformation for both frequency of GUS expression and GUS spots per callus. PIG-mediated transformation resulted in higher GUS expression compared to the Agrobacterium method. These results demonstrate that Dichanthium annulatum is amenable to Agrobacterium-mediated genetic transformation using a binary vector. |
Description: | Not Available |
ISSN: | Not Available |
Type(s) of content: | Research Paper |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | DNA and Cell Biology |
NAAS Rating: | 9.19 |
Volume No.: | 24(11) |
Page Number: | 670-679 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | Not Available |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/9162 |
Appears in Collections: | CS-DSBR-Publication |
Files in This Item:
File | Description | Size | Format | |
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invitro plant regeneration.pdf | 284.92 kB | Adobe PDF | View/Open |
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