Transformation of peanut (Arachis hypogaea L.): a non-tissue culture based approach for generating transgenic plants

Rohini, V. K.; Rao, K. Sambasiva

doi:10.1016/s0168-9452(99)00160-0

Cited by 98 publications

(58 citation statements)

References 19 publications

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“…K-134, transgenics were developed by a tissue culture independent Agrobacterium-mediated in planta transformation strategy. The in planta transformation protocols have been developed for transformation in many crop species, Arabidopsis thaliana (Feldmann and Marks 1987), soybean (Trick and Finer 1997), sunflower (Rao and Rohini 1999), safflower (Rohini and Rao 2000), rice (Supartana et al 2005), maize (Chumakov et al 2006) and cotton (Keshamma et al 2008). Earlier an in planta transformation protocol was developed in groundnut by Rohini and Rao (2000) and proved its reproducibility by Keshamma et al (2008).…”

Section: Discussionmentioning

confidence: 99%

“…The in planta transformation protocols have been developed for transformation in many crop species, Arabidopsis thaliana (Feldmann and Marks 1987), soybean (Trick and Finer 1997), sunflower (Rao and Rohini 1999), safflower (Rohini and Rao 2000), rice (Supartana et al 2005), maize (Chumakov et al 2006) and cotton (Keshamma et al 2008). Earlier an in planta transformation protocol was developed in groundnut by Rohini and Rao (2000) and proved its reproducibility by Keshamma et al (2008). In the present study, selection of stable transformants by employing the in planta transformation strategy in groundnut was ascertained by PCR analysis of the plants for selectable marker gene in T 1 generation, and for both marker gene and cry1AcF gene in T 2 generation.…”

Section: Discussionmentioning

confidence: 99%

“…Transformation and recovery of transformants In planta transformation protocol standardized by our group (Rohini and Rao 2000;Keshamma et al 2008;Kumar et al 2009) was followed to obtain groundnut transformants. The seedlings with just emerging plumule were infected by pricking at the shoot apical meristem with a sterile needle and subsequent immersion in the culture of Agrobacterium and incubated at 28°C, 40 rpm for 60 min.…”

Section: Methodsmentioning

confidence: 99%

“…To understand the efficacy of the chimeric Bt cry1AcF gene in groundnut, a tissue culture-independent in planta transformation strategy (Rohini and Rao 2000) was used. Transgenics were validated for the efficacy against S. litura.…”

Section: Introductionmentioning

confidence: 99%

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Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Keshavareddy

Rohini

Ramu

et al. 2013

Physiol Mol Biol Plants

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Large number of primary transgenic events were generated in groundnut by an Agrobacterium mediated, in planta transformation method to assess the efficacy of cry1AcF against the Spodoptera litura. generation demonstrated homozygous nature. This clearly proved that though there is considerable improvement in average mean % larval mortality in T 2 generation, the cry1AcF gene was effective against S. litura only to some extent.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Keshavareddy

Rohini

Ramu

et al. 2013

Physiol Mol Biol Plants

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“…After 4 h of induction, the germlings which were wounded by gently pricking the cotyledonary node at the embryo axis 3-4 times using a sterile sewing needle were immersed in the A.B broth and incubated at 28°C without agitation for one hour. Later,the germlings were treated with 500 μg ml− 1 of cefotaxime in liquid MS basal medium (Murashige and Skoog 1962) for 20 min (Rohini and Sankara Rao 2000) and then placed on moistened autoclaved soilrite (Keltec Industries, Bangalore) in 250 ml wide mouth bottles for further growth in the growth room maintained under standard conditions of 28°C, and 14 hday time (30 μmol m −2 s −1 ) . After 10-12 days the germlings were transferred to soilrite containing polythene bags and allowed to grow in the growth room.…”

Section: Methodsmentioning

confidence: 99%

Tissue culture independent transformation of the forage crop sunnhemp (Crotalaria juncea L.): an easy method towards generation of transgenics

Rao¹,

Agrawal²,

Mahmood

et al. 2011

Physiol Mol Biol Plants

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A transformation system which is free of in vitro plant regeneration following Agrobacterium infection is established for the forage legume, Sunnhemp (Crotalaria juncea L.) where in the entire embryo axis of the germinating seed was used as the target tissue for transformation. After standardization of transformation conditions, the cotyledonary node of the embryo axis was infected with Agrobacterium host LBA 4404 harboring the recombinant vector pCAMBIA 2301. The bivalent 1D gene of the two major foot and mouth disease virus (FMDV) serotypes 'O' and 'A 22 ' and the neomycin phosphotransferase (nptII) gene were used as the markers for optimization of the protocol. The embryo axes were pricked randomly on the cotyledonary node and co-cultivated with Agrobacterium. The germlings were then allowed to grow under standard growth room conditions in to mature fertile plants. 60 T 0 plants were established from 3 separate experiments. Three hundred seeds from the 60 T 0 plants were sown to raise the T 1 generation of which 180 were analyzed for integration of bivalent FMDV gene 1D "O" and "A 22 " and the nptII gene. Eighteen out of these 180 plants amplified both the marker genes. Two independent transgenic lines 24 and 37, showed elevated levels of expression of 12 μg and 8 μg (per gm of fresh leaf) of the bivalent ID antigen "O" and "A 22 " . The results showed that the transformation efficiency was 3 %. To the best of our knowledge, this is the first successful attempt of Agrobacterium tumefaciens mediated transformation of Sunnhemp. The protocol can generate whole plant transformants with relative ease and should be compatible to all genotypes of Sunnhemp.

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Plant Transformation Technology:Agrobacterium‐Mediated Transformation

Komari

Ishida

Hiei

2004

Handbook of Plant Biotechnology

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The soil phytopathogen Agrobacterium tumefaciens induces tumours, known as crown galls, mainly on dicotyledonous plants. Such tumours are generated by a complex, multi‐step transformation process. Another species, A. rhizogenes , causes hairy root disease in higher plants via an identical process. Agrobacterium has been utilised for the transfer of genes to dicotyledonous plants. Now, monocotyledonous plants are routinely transformed by Agrobacterium despite the fact that these plants, including important cereals, were thought until recently to be outside the range of this technology. Most of the common transformation methods heavily depend on tissue culture technology and we refer to such methods as in vitro transformation. The only case of a routine tissue culture free method is the ‘ in planta ’ transformation of Arabidopsis thaliana . Numerous critical factors are involved in both approaches. In transformation in vitro , key factors include choice of vectors and bacterial strains, types of plant tissues to be infected, procedures of preparation the tissues, protocols of infection and co‐cultivation, methods for subsequent culture and selection of transformed cells, antibiotics to remove infecting bacteria, techniques for regeneration of transgenic plants and genotypes of plants. It is our opinion that the type and quality of the starting material is the most important one among them. The capacity to serve as a host plant for crown gall disease is no longer a prerequisite for a host of vector systems based on Agrobacterium . The real prerequisite for in vitro transformation is the availability of technology for dedifferentiation of tissues and regeneration of plants in a given species. On the other hand, the biological processes involved in in planta transformation are yet to be elucidated. Recent evidence suggested that ovules are the primary target, and further basic understanding is likely to help extend the number of species transformed by the method. The plant species that are routinely transformed by Agrobacterium are expanding rapidly. Some gymnosperms, several forest trees and fruit trees, various legumes, and cereal and non‐cereal monocotyledons, which were once considered very recalcitrant, are now in a long list of transformable plants. Arabidopsis , tobacco and rice are the top three species that were transformed during the last two years. Although it is only half a dozen years since the current procedure of rice transformation mediated by Agrobacterium was published, the economic importance and the accumulation of genomic information has made rice the species of focus in plant biotechnology. The advantages of Agrobacterium ‐mediated transformation include the transfer of pieces of DNA with defined ends and minimal rearrangement, the transfer of relatively large segments of DNA, the integration of small numbers of copies of genes into plant chromosomes and the high quality and fertility of transgenic plants. However, transformation does not always produce such ‘clean’ events. Formation of repeats of transgenes, certain rearrangements, integration of non‐target DNA segments and unstable expression of transgenes are among the complications. Although the majority of transgenic plants usually appear ‘good’ in a particular test, accumulation of ‘dropped’ plants is significant after multiple rounds of characterisation and screening. Therefore, further improvement in each of steps is highly desired. Application of the gene transfer mediated by Agrobacterium is further expanding. Transient expression of genes delivered by Agrobacterium is now a useful tool in the study of promoters and gene function. Vectors specifically designed to carry very large segments of DNA have been developed and extensively tested. Insertional mutagenesis by DNA transferred by Agrobacterium is a routine technique in genomics study in Arabidopsis and rice. Methods for targeted integration of transgenes to genomes of higher plants have been drawing considerable attention. Various technologies for the production of ‘selection marker free’ transgenic plants are now in place for better public acceptance of biotechnology products.

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Transformation of peanut (Arachis hypogaea L.): a non-tissue culture based approach for generating transgenic plants

Cited by 98 publications

References 19 publications

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Transgenics in groundnut (Arachis hypogaea L.) expressing cry1AcF gene for resistance to Spodoptera litura (F.)

Tissue culture independent transformation of the forage crop sunnhemp (Crotalaria juncea L.): an easy method towards generation of transgenics

Plant Transformation Technology:Agrobacterium‐Mediated Transformation

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