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Antisense expression of a gene encoding a calcium-binding protein in transgenic tobacco leads to altered morphology and enhanced chlorophyll

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Abstract

Entamoeba histolytica contains a novel calcium-binding protein like calmodulin, which was discovered earlier, and we have reported the presence of its homologue(s) and a dependent protein kinase in plants. To understand the functions of these in plants, a cDNA encoding a calcium-binding protein isolated from Entamoeba histolytica (EhCaBP) was cloned into vector pBI121 in antisense orientation and transgenic tobacco plants were raised. These plants showed variation in several phenotypic characters, of which two distinct features, more greenness and leaf thickness, were inherited in subsequent generations. The increase in the level of total chlorophyll in different plants ranged from 60% to 70%. There was no major change in chloroplast structure and in the protein level of D1, D2, LHCP and RuBP carboxylase. These morphological changes were not seen in antisense calmodulin transgenic tobacco plants, nor was the calmodulin level altered in EhCaBP antisense plants.

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Abbreviations

ABA:

abscisic acid

CaM:

calmodulin

CBl:

calcineurin B-like proteins

CDPK:

calcium-dependent protein kinase

CIPK:

CBL interacting protein kinases

dZR:

dihydroxyzeatin riboside

EF:

elongation factor motif

EhCaBP:

Entamoeba histolytica calcium-binding protein

GUS:

beta-glucuronidase

IgG:

immunoglobulin G

iPA:

isopentenyl adenosine

LHCP:

light harvesting chlorophyll binding project

LSU:

large subunit

PCR:

polymerase chain reaction

PKS11:

protein kinase 11

PS:

photosystem

RuBP:

ribulose-1,5-bisphosphate carboxylase/oxygenase

SOS2:

salt overly sensitive 2

TLC:

thin layer chromatography

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Author notes

  1. Girdhar K Pandey & Amita Pandey

    Present address: Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720, USA

Authors and Affiliations

  1. International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, 110 067, India

    Girdhar K Pandey, Vanga Siva Reddy & Sudhir K. Sopory

  2. School of Life Sciences, Jawaharlal Nehru University, New Delhi, India

    Amita Pandey, Alok Bhattacharya & Kailash C Upadhyaya

  3. Department of Botany, University of Delhi, New Delhi, India

    Renu Deswal

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  1. Girdhar K Pandey
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  2. Amita Pandey
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  3. Vanga Siva Reddy
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Correspondence to Girdhar K Pandey.

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The results of this paper have been granted US Patent No. 6,791,009.

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Pandey, G.K., Pandey, A., Reddy, V.S. et al. Antisense expression of a gene encoding a calcium-binding protein in transgenic tobacco leads to altered morphology and enhanced chlorophyll. J Biosci 32, 251–260 (2007). https://doi.org/10.1007/s12038-007-0025-0

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  • DOI: https://doi.org/10.1007/s12038-007-0025-0

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