Shoot apical meristem injection

A novel and efficient method to obtain transformed cucumber plants

P. Baskaran, V. Soós, E. Balázs, J. Van Staden

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The objective of this study was to develop a simple and efficient system for the genetic transformation of cucumber (Cucumis sativus L.). This study investigated the factors influencing the efficiency of gene transfer by microinjection of the shoot apical meristem (SAM) with Agrobacterium shooter strain. The SAM microinjected with Agrobacterium cells containing binary plasmid was incubated in co-cultivation medium without plant growth regulators (PGRs) for 2days in the dark and subsequently transferred to solid bacterial elimination shoot regeneration medium containing MS salts plus 20gl-1 glucose, 8gl-1 agar and 300mgl-1 cefotaxime. Developed transgenic shoots were screened in selection medium containing the antibiotic kanamycin (Km). The efficiency of gene transfer using various germination media and infection conditions was analyzed by root growth in the presence of Km. The present investigations show that the maximum efficiency of gene transfer was sixty-five percent by Km-resistant plants. In the putative transgenic plants the presence of the transgene was confirmed by polymerase chain reaction (PCR). This transformation system was reproducible, suggesting that it could be adopted for other plant species.

Original languageEnglish
Pages (from-to)210-215
Number of pages6
JournalSouth African Journal of Botany
Volume103
DOIs
Publication statusPublished - Mar 1 2016

Fingerprint

shoot meristems
apical meristems
kanamycin
gene transfer
cucumbers
injection
Agrobacterium
cefotaxime
shoots
genetic transformation
Cucumis sativus
plant growth substances
transgenes
transgenic plants
root growth
plasmids
agar
polymerase chain reaction
methodology
antibiotics

Keywords

  • Agrobacterium shooter strain
  • Cucumber
  • Gene transfer
  • Microinjection
  • Shoot apical meristem

ASJC Scopus subject areas

  • Plant Science

Cite this

Shoot apical meristem injection : A novel and efficient method to obtain transformed cucumber plants. / Baskaran, P.; Soós, V.; Balázs, E.; Van Staden, J.

In: South African Journal of Botany, Vol. 103, 01.03.2016, p. 210-215.

Research output: Contribution to journalArticle

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