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RETRACTED ARTICLE: High-efficiency Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) and regeneration of insect-resistant transgenic plants

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This article was retracted on 02 August 2013

Abstract

To develop an efficient genetic transformation system of chickpea (Cicer arietinum L.), callus derived from mature embryonic axes of variety P-362 was transformed with Agrobacterium tumefaciens strain LBA4404 harboring p35SGUS-INT plasmid containing the uidA gene encoding β-glucuronidase (GUS) and the nptII gene for kanamycin selection. Various factors affecting transformation efficiency were optimized; as Agrobacterium suspension at OD600 0.3 with 48 h of co-cultivation period at 20°C was found optimal for transforming 10-day-old MEA-derived callus. Inclusion of 200 μM acetosyringone, sonication for 4 s with vacuum infiltration for 6 min improved the number of GUS foci per responding explant from 1.0 to 38.6, as determined by histochemical GUS assay. For introducing the insect-resistant trait into chickpea, binary vector pRD400-cry1Ac was also transformed under optimized conditions and 18 T0 transgenic plants were generated, representing 3.6% transformation frequency. T0 transgenic plants reflected Mendelian inheritance pattern of transgene segregation in T1 progeny. PCR, RT-PCR, and Southern hybridization analysis of T0 and T1 transgenic plants confirmed stable integration of transgenes into the chickpea genome. The expression level of Bt-Cry protein in T0 and T1 transgenic chickpea plants was achieved maximum up to 116 ng mg−1 of soluble protein, which efficiently causes 100% mortality to second instar larvae of Helicoverpa armigera as analyzed by an insect mortality bioassay. Our results demonstrate an efficient and rapid transformation system of chickpea for producing non-chimeric transgenic plants with high frequency. These findings will certainly accelerate the development of chickpea plants with novel traits.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

Bt:

Bacillus thuringiensis

CIM:

Callus induction medium

Cry:

Crystal protein

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MEA:

Mature embryonic axes

MS:

Murashige and Skoog medium

nptII:

Neomycin phosphotransferase

PGR:

Plant growth regulators

uidA :

β-Glucouronidase

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Acknowledgments

We are thankful to Council of Scientific and Industrial Research, New Delhi for providing funds and research fellowships. We thankfully acknowledge Prof. I. Altosaar, Department of Biochemistry, University of Ottawa, Ottawa, Canada for providing synthetic modified Bt-cry1Ac gene. This work was carried out under the CSIR Network Project NWP0003 and OLP0031.

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Authors and Affiliations

  1. Plant Transgenic Lab, National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, India

    Meenakshi Mehrotra, Indraneel Sanyal & D. V. Amla

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  1. Meenakshi Mehrotra
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  2. Indraneel Sanyal
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  3. D. V. Amla
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Correspondence to Meenakshi Mehrotra.

Additional information

Communicated by P. Kumar.

This article has been retracted on the demand of the authors because of error in one figure panel. Consequently, the data is not unambiguous. All of the authors have agreed with the retraction notice and sincerely regret the inconvenience that this retraction causes to PCR and its readership.

The retraction note to this article can be found online at http://dx.doi.org/10.1007/s00299-013-1485-3.

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Mehrotra, M., Sanyal, I. & Amla, D.V. RETRACTED ARTICLE: High-efficiency Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) and regeneration of insect-resistant transgenic plants. Plant Cell Rep 30, 1603–1616 (2011). https://doi.org/10.1007/s00299-011-1071-5

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  • DOI: https://doi.org/10.1007/s00299-011-1071-5

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