Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 112, Issue 2, pp 217–226 | Cite as

Agrobacterium rhizogenes mediated transformation of the medicinal plant Decalepis arayalpathra and production of 2-hydroxy-4-methoxy benzaldehyde

  • C. G. Sudha
  • T. V. Sherina
  • V. P. Anu Anand
  • J. V. Reji
  • P. Padmesh
  • E. V. Soniya
Original Paper

Abstract

Agrobacterium rhizogenes mediated transformation of Decalepis arayalpathra, an ethnomedicinal plant, was achieved by infecting juvenile hypocotyl explants with different strains, including A4, MTCC 532, TR105 and LBA 5402. Hypocotyl explants induced hairy roots at a higher frequency (53.2 ± 0.3 %) than cotyledons (32.1 ± 0.2 %) when infected with the most virulent strain TR105. The explants co-cultivated 48 h in half-strength salts and vitamins of Murashige and Skoog basal medium (half-MSB) induced hairy roots either directly from the wounds or followed by the formation of gall like structures. Irrespective of the explants, the strain MTCC 532 induced callus alone. The root initials on the galls proliferated vigorously and elongated more rapidly when they were segmented and subcultured on half-MSB medium than the proliferation and elongation of directly emerged roots. The established hairy roots showed intermittent gall formation which was the active sites for hairy roots induction. The molecular evidence of rol A and rol C gene integration was confirmed by PCR amplification and southern blot hybridization. Growth of the hairy roots was undertaken by measuring root growth unit after culturing root tips in half-MSB solid medium and determined fresh weight/dry weight/conductivity during time-course study in shake flask cultures. The maximum biomass and accumulation of the root specific compound, 2-hydroxy-4-methoxy benzaldehyde (MBALD) (0.22 % dry weight), was recorded at the 6th week of growth, which was more than that observed in normal root cultures (0.16 % dry weight).

Keywords

Agrobacterium rhizogenes Decalepis arayalpathra 2-hydroxy-4-methoxy benzaldehyde MBALD 

Abbreviation

dw

Dry weight

fw

Fresh weight

HPLC

High Performance Liquid Chromatography

IBA

Indole-3-butyric acid

MBALD

2-Hydroxy-4-methoxy benzaldehyde

MSB

Murashige and Skoog basal medium

NAA

α-Naphthaleneacetic acid

RGU

Root growth unit

TLC

Thin Layer Chromatography

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • C. G. Sudha
    • 1
  • T. V. Sherina
    • 1
  • V. P. Anu Anand
    • 1
  • J. V. Reji
    • 1
    • 3
  • P. Padmesh
    • 1
  • E. V. Soniya
    • 2
  1. 1.Biotechnology and Bioinformatics DivisionJawaharlal Nehru Tropical Botanic Garden and Research InstitutePalode, ThiruvananthapuramIndia
  2. 2.Plant Molecular Biology DivisionRajiv Gandhi Centre for BiotechnologyThiruvananthapuramIndia
  3. 3.Department of BiotechnologySathyabama UniversityChennaiIndia

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