Expression of a synthetic cry1AcF gene in transgenic Pigeon pea confers resistance to Helicoverpa armigera

Pigeon pea is an important legume. Yield losses due to insect pests are enormous in the cultivation of this crop. Expression of cry proteins has led to increased resistance to pests in several crops. We report in this paper, expression of a chimeric cry1AcF (encoding cry1Ac and cry1F domains) gene in transgenic pigeon pea and its resistance towards Helicoverpa armigera. PCR, Southern hybridization, RT‐PCR and Western analysis confirmed stable integration and expression of the cry1AcF gene in pigeon pea transgenics. When screened for efficacy of the transformants for resistance against H. armigera, the transgenics showed not only high mortality of the larva but could also resist the damage caused by the larvae. Analysis for the stable integration, expression and efficacy of the transgenics resulted in the identification of four T3 plants arising from two T1 backgrounds as highly promising. The results demonstrate potentiality of the chimeric cry1AcF gene in developing H. armigera‐resistant pigeon pea.

[1]  G. D..  Tissue Culture Approaches to Pigeonpea Improvement , 2011 .

[2]  S. Sundaresha,et al.  Enhanced protection against two major fungal pathogens of groundnut, Cercospora arachidicola and Aspergillus flavus in transgenic groundnut over-expressing a tobacco β 1–3 glucanase , 2010, European Journal of Plant Pathology.

[3]  R. Sreevathsa,et al.  Towards crop improvement in bell pepper (Capsicum annuum L.): Transgenics (uid A::hpt II) by a tissue-culture-independent Agrobacterium-mediated in planta approach , 2009 .

[4]  R. Sreevathsa,et al.  In planta transformation of pigeon pea: a method to overcome recalcitrancy of the crop to regeneration in vitro , 2008, Physiology and Molecular Biology of Plants.

[5]  K. Sharma,et al.  Agrobacterium-mediated production of transgenic pigeonpea (Cajanus cajan L. Millsp.) expressing the synthetic BT cry1Ab gene , 2006, In Vitro Cellular & Developmental Biology - Plant.

[6]  James B. Hicks,et al.  A plant DNA minipreparation: Version II , 1983, Plant Molecular Biology Reporter.

[7]  H. Upadhyaya,et al.  Exploitation of wild relatives of pigeonpea and chickpea for resistance to Helicoverpa armigera , 2007 .

[8]  Tsutomu Shimizu,et al.  Development of simple and efficient in Planta transformation method for wheat (Triticum aestivum L.) using Agrobacterium tumefaciens. , 2006, Journal of bioscience and bioengineering.

[9]  I. Volokhina,et al.  Agrobacterium-mediated in planta transformation of maize via pistil filaments , 2006, Russian Journal of Genetics.

[10]  N. Baisakh,et al.  Translational fusion hybrid Bt genes confer resistance against yellow stem borer in transgenic elite vietnamese rice (Oryza sativa L.) cultivars , 2006 .

[11]  P. Kirti,et al.  Agrobacterium-mediated genetic transformation of pigeon pea (Cajanus cajan (L.) Millsp.) using embryonal segments and development of transgenic plants for resistance against Spodoptera , 2005 .

[12]  H. Sharma,et al.  Genetic Transformation of Crops for Insect Resistance: Potential and Limitations , 2004 .

[13]  R. D. de Maagd,et al.  A hybrid Bacillus thuringiensis delta-endotoxin gives resistance against a coleopteran and a lepidopteran pest in transgenic potato. , 2002, Plant biotechnology journal.

[14]  N. Baisakh,et al.  Pyramiding transgenes for multiple resistance in rice against bacterial blight, yellow stem borer and sheath blight , 2002, Theoretical and Applied Genetics.

[15]  A. Shelton,et al.  Broccoli plants with pyramided cry1Ac and cry1C Bt genes control diamondback moths resistant to Cry1A and Cry1C proteins , 2002, Theoretical and Applied Genetics.

[16]  P. Kumar,et al.  Insecticidal Proteins of Bacillus Thuringiensis and Their Application in Agriculture , 2002 .

[17]  Z. W. Shappley,et al.  Bollgard II efficacy: quantification of total lepidopteran activity in a 2-gene product. , 2000 .

[18]  R. Frutos,et al.  Managing Insect Resistance to Plants Producing Bacillus thuringiensis Toxins , 1999 .

[19]  Kumar,et al.  Efficacy of lepidopteran specific delta-endotoxins of bacillus thuringiensis against helicoverpa armigera , 1998, Journal of invertebrate pathology.

[20]  K. Krishnamurthy,et al.  Plant regeneration in pigeonpea [Cajanus cajan (L.) Millsp.] by organogenesis , 1998, Plant Cell Reports.

[21]  A. Mandaokar,et al.  SYNERGISTIC EFFECT OF CRY1AC AND CRY1F DELTA -ENDOTOXONS OF BACILLUS THURINGIENSIS ON COTTON BOLLWORM, HELICOVERPA ARMIGERA , 1998 .

[22]  K. Feldmann,et al.  T-DNA insertion mutagenesis in Arabidopsis: going back and forth. , 1997, Trends in genetics : TIG.

[23]  Y. Nene,et al.  Pigeonpea: geography and importance. , 1990 .

[24]  D. G. Faris,et al.  Tissue Culture Approaches to Pigeonpea Improvement , 1988 .