Genetic transformation of cultivated sesame (Sesamum indicum L. cv Rama) through particle bombardment using 5-day-old apical, meristematic tissues of germinating seedlings

Jagannath Bhattacharyya, Anirban Chakraborty, Joy Mitra, Saikat Chakraborty, Subrata Pradhan, Anulina Manna, Narattam Sikdar, Soumitra Kumar Sen

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

An in vitro plant generation and genetic transformation protocol was established in sesame (Sesamum indicum L. cv Rama) through biolistic particle gun bombardment. 5-day-old apical, meristematic tissues of in vitro-germinating seedlings were used as explants. 10–15 Multiple shoots were generated from each explant using Murashige and Skoog basal medium containing 18.0 µM benzylamino purine and 5.37 µM naphthalene acetic acid. Four independent sets of transformation were carried out and each set consisted of three independent experiments each comprising three replications with 30 explants per replication. A synthetically designed bialaphos resistance gene (bar) was used for transformation. The positive transformants containing the bar gene were selected in growth medium containing 2.5 mg/L bialaphos. Green shoots recovered from bombarded explants were subjected to root development on Murashige and Skoog basal medium containing 5.37 µM naphthalene acetic acid. The rooted shoots were established in soil and grown to maturity in greenhouse. Polymerase chain reaction (PCR), Southern and reverse-transcription PCR, real-time quantitative PCR, western blot and enzymatic assay of four putative transformants from independent sets provided evidence for full-length gene integration as well as high level expression of the transgene. Analysis of the T1 plants revealed a stable inheritance of the transgene through the progenies. This is the first report of biolistic mediated stable transformation of sesame and should pave the way for future genetic engineering strategies to be employed for improvement of this very important oil-seed crop.

Original languageEnglish (US)
Pages (from-to)455-466
Number of pages12
JournalPlant Cell, Tissue and Organ Culture
Volume123
Issue number3
DOIs
StatePublished - Dec 1 2015
Externally publishedYes

Fingerprint

Sesamum indicum
genetic transformation
explants
biolistics
seedlings
naphthaleneacetic acid
transgenes
shoots
genes
oilseed crops
purines
genetic engineering
inheritance (genetics)
quantitative polymerase chain reaction
Western blotting
culture media
polymerase chain reaction
reverse transcriptase polymerase chain reaction
greenhouses
tissues

Keywords

  • Bar gene
  • Bialaphos
  • Particle bombardment
  • Sesame transformation
  • Transgenic plants

ASJC Scopus subject areas

  • Horticulture

Cite this

Genetic transformation of cultivated sesame (Sesamum indicum L. cv Rama) through particle bombardment using 5-day-old apical, meristematic tissues of germinating seedlings. / Bhattacharyya, Jagannath; Chakraborty, Anirban; Mitra, Joy; Chakraborty, Saikat; Pradhan, Subrata; Manna, Anulina; Sikdar, Narattam; Sen, Soumitra Kumar.

In: Plant Cell, Tissue and Organ Culture, Vol. 123, No. 3, 01.12.2015, p. 455-466.

Research output: Contribution to journalArticle

Bhattacharyya, Jagannath ; Chakraborty, Anirban ; Mitra, Joy ; Chakraborty, Saikat ; Pradhan, Subrata ; Manna, Anulina ; Sikdar, Narattam ; Sen, Soumitra Kumar. / Genetic transformation of cultivated sesame (Sesamum indicum L. cv Rama) through particle bombardment using 5-day-old apical, meristematic tissues of germinating seedlings. In: Plant Cell, Tissue and Organ Culture. 2015 ; Vol. 123, No. 3. pp. 455-466.
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AU - Chakraborty, Saikat

AU - Pradhan, Subrata

AU - Manna, Anulina

AU - Sikdar, Narattam

AU - Sen, Soumitra Kumar

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