| Title |
Optimization of Agrobacterium mediated genetic transformation of cotyledonary node explants of Vigna radiata
|
|---|---|
| Published in |
SpringerPlus, December 2012
|
| DOI | 10.1186/2193-1801-1-59 |
| Pubmed ID | |
| Authors |
Sushil Kumar Yadav, Sweety Katikala, Varalaxmi Yellisetty, Annapurna Kannepalle, Jyothi Lakshmi Narayana, Vanaja Maddi, Maheswari Mandapaka, Arun Kumar Shanker, Venkateswarlu Bandi, Kirti Pulugurtha Bharadwaja |
| Abstract |
A reproducible and highly efficient protocol for genetic transformation mediated by Agrobacterium has been established for greengram (Vigna radiata L. Wilczek). Double cotyledonary node (DCN) explants were inoculated with Agrobacterium tumefaciens strain LBA 4404 harboring a binary vector pCAMBIA 2301 containing neomycin phosphotransferase (npt II) gene as selectable marker, β-glucuronidase (GUS) as a reporter (uidA) gene and annexin 1 bj gene. Important parameters like optical density of Agrobacterium culture, culture quantity, infection medium, infection and co-cultivation time and acetosyringone concentration were standardized to optimize the transformation frequency. Kanamycin at a concentration of 100 mg/l was used to select transformed cells. Transient and stable GUS expressions were studied in transformed explants and regenerated putative plants, respectively. Transformed shoot were produced on regeneration medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime and rooted on ½ MS medium. Transient and constitutive GUS expression was observed in DCN explants and different tissues of T(0) and T(1) plants. Rooted T(0) and T(1) shoots confirming Polymerase Chain Reaction (PCR) positive for npt II and annexin 1bj genes were taken to maturity to collect the seeds. Integration of annexin gene into the greengram genome was confirmed by Southern blotting. |
Mendeley readers
The data shown below were compiled from readership statistics for 55 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| India | 1 | 2% |
| Unknown | 54 | 98% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 16 | 29% |
| Student > Master | 9 | 16% |
| Researcher | 7 | 13% |
| Student > Bachelor | 4 | 7% |
| Student > Doctoral Student | 3 | 5% |
| Other | 8 | 15% |
| Unknown | 8 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 30 | 55% |
| Biochemistry, Genetics and Molecular Biology | 8 | 15% |
| Engineering | 2 | 4% |
| Social Sciences | 1 | 2% |
| Unknown | 14 | 25% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 27 November 2013.
All research outputs
#17,673,866
of 22,689,790 outputs
Outputs from SpringerPlus
#1,202
of 1,851 outputs
Outputs of similar age
#208,496
of 278,728 outputs
Outputs of similar age from SpringerPlus
#18
of 48 outputs
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We're also able to compare this research output to 48 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 60% of its contemporaries.