High efficiency transformation protocol for three Indian cotton varieties via Agrobacterium tumefaciens

V. V. Satyavathi, V. Prasad, B. Gita Lakshmi, G. Lakshmi Sita

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

A protocol for consistent production of transgenic cotton plants in three Indian varieties was established utilizing Agrobacterium-mediated transformation. Shoot tip explants were transformed by cocultivation with Agrobacterium tumefaciens strain LBA 4404. The strain harbors a binary vector pBAL2 carrying the reporter gene β-glucuronidase intron (GUS-INT) and the marker gene neomycin phosphotransferase (NPTII). Regeneration potential of explants or different hormones was studied in detail. Among the different combinations of BAP and NAA tested, 0.1 mg 1-1 of BAP and NAA in the medium influenced efficient regeneration of shoots by organogenesis. Shoot bud proliferation and elongation was achieved in 3-4 weeks time on medium supplemented with GA3. The putatively transformed shoots were harvested and placed for rooting on medium containing IBA and 75mg 1-1 kanamycin. Transgenic plants were recovered in 12-16 weeks from the time of gene transfer to establishment in pots. Molecular analysis of the field established plantlets was carried to confirm the transgenic nature. The presence of GUS and NPTII genes in the transgenic plants was verified by histochemical GUS assay and polymerase chain reaction (PCR) analysis, respectively. Integration of T-DNA into the genome of putative transgenics was further confirmed by Southern blot analysis. A total of 70-75 transgenic plants were raised in pots. Progeny analysis of these plants showed a classical Mendelian pattern of inheritance.

Original languageEnglish
Pages (from-to)215-223
Number of pages9
JournalPlant Science
Volume162
Issue number2
DOIs
Publication statusPublished - 11-02-2002
Externally publishedYes

Fingerprint

Agrobacterium tumefaciens
Genetically Modified Plants
Agrobacterium radiobacter
transgenic plants
cotton
genetically modified organisms
shoots
explants
Regeneration
kanamycin kinase
Kanamycin Kinase
kanamycin
Genes
organogenesis
Gossypium
Agrobacterium
gene transfer
reporter genes
Inheritance Patterns
Southern blotting

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

Cite this

Satyavathi, V. V. ; Prasad, V. ; Gita Lakshmi, B. ; Lakshmi Sita, G. / High efficiency transformation protocol for three Indian cotton varieties via Agrobacterium tumefaciens. In: Plant Science. 2002 ; Vol. 162, No. 2. pp. 215-223.
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High efficiency transformation protocol for three Indian cotton varieties via Agrobacterium tumefaciens. / Satyavathi, V. V.; Prasad, V.; Gita Lakshmi, B.; Lakshmi Sita, G.

In: Plant Science, Vol. 162, No. 2, 11.02.2002, p. 215-223.

Research output: Contribution to journalArticle

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