Transgenic indica Rice Variety Pusa Basmati 1 Constitutively Expressing a Rice Chitinase Gene Exhibits Enhanced Resistance to Rhizoctonia solani

  • G. Sridevi
  • N. Sabapathi
  • P. Meena
  • R. Nandakumar
  • R. Samiyappan
  • S. Muthukrishnan
  • K. Veluthambi
Article

Abstract

Agrobacterium-mediated transformation of an elite indica rice variety, Pusa Basmati 1, was performed using LBA4404 (pSB1, pMKU-RF2) that harbours a rice chitinase gene (chi11) under the control of the maize ubiquitin (Ubi1) promoter-intron. Right border (gus) and left border (hph) flanking sequences and the transgene (chi11) in the middle of the T-DNA were used as probes in Southern analysis. Out of eleven independent T0 plants regenerated, three had single copy T-DNA insertions and eight had multiple T-DNA insertions. Nine T0 plants carried the complete T-DNA with the chitinase transgene. Two T0 plants did not carry chi11, though they had other T-DNA portions. Three plants harbouring single copy insertions and one plant harbouring two inserted copies were analyzed in detail. A segregation ratio of 3:1, reflecting T-DNA insertion at a single locus, was observed in the progeny of all the four T0 plants. Northern and western blot analyses of T1 plants revealed constitutive expression of chitinase at high levels. Bioassays of T1 plants indicated enhanced resistance to the sheath blight pathogen, Rhizoctonia solani, in comparison to control plants. A homozygous transgenic line was established from one T0 line, which exhibited the maximum resistance to R. solani.

Keywords

sheath blight Rhizoctonia solani chitinase transgenic rice Agrobacterium 

Abbreviations

AS

acetosyringone

BCIP

5-bromo-4-chloro-3-indolyl phosphate

chi11

rice chitinase gene

CI

callus-induction

2,4-D

2,4-dichlorophenoxyacetic acid

gus

β-glucuronidase

hyg

hygromycin

hph

hygromycin phosphotransferase

int-gus

introncontaining gus gene

LB

left T-DNA border

MS

Murashige and Skoog

NAA

α-naphthaleneacetic acid

NBT

nitroblue tetrazolium

PB1

Pusa Basmati 1

PR

pathogenesis-related

RM

regeneration medium

RB

right T-DNA border

TTBS

Tween-Tris buffered saline

Ubi1

maize ubiquitin promoter-intron

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • G. Sridevi
    • 1
  • N. Sabapathi
    • 1
  • P. Meena
    • 1
  • R. Nandakumar
    • 2
  • R. Samiyappan
    • 2
  • S. Muthukrishnan
    • 3
  • K. Veluthambi
    • 1
  1. 1.Department of Plant Biotechnology, School of BiotechnologyMadurai Kamaraj UniversityMaduraiTamil Nadu, India
  2. 2.Department of Plant PathologyTamil Nadu Agricultural UniversityCoimbatoreTamil Nadu, India
  3. 3.Department of BiochemistryKansas State UniversityManhattanUSA

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