An optimized Agrobacterium -mediated transformation procedure for Phaseolus acutifolius A. Gray

Clercq, Janniek De; Zambre, Mukund; Montagu, Marc Van; Dillen, Willy; Angenon, Geert

doi:10.1007/s00299-002-0518-0

Cited by 73 publications

(45 citation statements)

References 49 publications

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“…Similar affirmative effects has been reported in several leguminous species such as Pisum sativum (Svabova and Griga 2008), Glycine max (Santarem et al 1998;Ko and Korban 2004), Vigna angularis (Yamada et al 2001), Phaseolus vulgaris (De Clercq et al 2002), Vigna mungo (Saini et al 2003;Muruganantham et al 2007), Vigna unguiculata (Popelka et al 2006) and Vigna radiata (Sonia et al 2007). Acetosyringone increases transformation potential of Agrobacterium strain by enhancing vir functions during transformation (Stachel et al 1986) and has been used in several other plant species also (Atkinson and Gardner 1991;Kaneyashi et al 1994).…”

Section: Optimization Of Transformation Protocolsupporting

confidence: 72%

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

Sainger

Chaudhary

Dahiya

et al. 2015

Physiol Mol Biol Plants

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An efficient, rapid and direct multiple shoot regeneration system amenable to Agrobacterium-mediated transformation from primary leaf with intact petiole of blackgram (Vigna mungo) is established for the first time. The effect of the explant type and its age, type and concentration of cytokinin and auxin either alone or in combination and genotype on multiple shoot regeneration efficiency and frequency was optimized. The primary leaf explants with petiole excised from 4-day-old seedlings directly developed multiple shoots (an average of 10 shoots/ explant) from the cut ends of the petiole in 95 % of the cultures on MSB (MS salts and B 5 vitamins) medium containing 1.0 μM 6-benzylaminopurine. Elongated (2-3 cm) shoots were rooted on MSB medium with 2.5 μM indole-butyric acid and resulted plantlets were hardened and established in soil, where they resumed growth and reached maturity with normal seed set. The regenerated plants were morphologically similar to seed-raised plants and required 8 weeks time from initiation of culture to establish them in soil. The regeneration competent cells present at the cut ends of petiole are fully exposed and are, thus, easily accessible to Agrobacterium, making this plant regeneration protocol amenable for the production of transgenic plants. The protocol was further successfully used to develop fertile transgenic plants of blackgram using Agrobacterium tumefaciens strain EHA 105 carrying a binary vector pCAMBIA2301 that contains a neomycin phosphotransferase gene (nptII) and a β-glucuronidase (GUS) gene (uidA) interrupted with an intron.The presence and integration of transgenes in putative T 0 plants were confirmed by polymerase chain reaction (PCR) and Southern blot hybridization, respectively. The transgenes were inherited in Mendelian fashion in T 1 progeny and a transformation frequency of 1.3 % was obtained. This protocol can be effectively used for transferring new traits in blackgram and other legumes for their quantitative and qualitative improvements.

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Section: Optimization Of Transformation Protocolsupporting

confidence: 72%

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

Sainger

Chaudhary

Dahiya

et al. 2015

Physiol Mol Biol Plants

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“…The use of Agrobacterium tumefaciens as a vector for legume transformation was an important breakthrough. Both micro-particle bombardment (Gulati et al, 2002; and A. tumefaciens (De Clercq et al, 2002;Li et al, 2004) have been used for DNA delivery into either embryogenic or organogenic cultures. Transformation has been generally based on infection by A. tumefaciens, although A. rhizogenes is also used for transformation of some species to produce composite plants with hairy roots or hairy root cultures (BoissonDernier et al, 2001;Stiller et al, 1997;Wu & VanEtten, 2004).…”

Section: Genetic Transformationmentioning

confidence: 99%

Biotechnology approaches to overcome biotic and abiotic stress constraints in legumes

et al. 2006

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Biotic and abiotic stresses cause significant yield losses in legumes and can significantly affect their productivity. Biotechnology tools such as marker-assisted breeding, tissue culture, in vitro mutagenesis and genetic transformation can contribute to solve or reduce some of these constraints. However, only limited success has been achieved so far. The emergence of "omic" technologies and the establishment of model legume plants such as Medicago truncatula and Lotus japonicus are promising strategies for understanding the molecular genetic basis of stress resistance, which is an important bottleneck for molecular breeding. Understanding the mechanisms that regulate the expression of stress-related genes is a fundamental issue in plant biology and will be necessary for the genetic improvement of legumes. In this review, we describe the current status of biotechnology approaches in relation to biotic and abiotic stresses in legumes and how these useful tools could be used to improve resistance to important constraints affecting legume crops.

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“…For qualitative assay, the area of GUS expression and intensity of the blue colour at each spot were given due weightage so as to give a holistic picture of the effects of the various treatments tested. Through visual observations, distinction could be made between small spots (Ͻ0.5 mm in diameter) representing one or few GUS expressing cells, and large spots (Ն1 mm in diameter), representing a complete cell cluster expressing the GUS gene (De Clercq et al 2002). Based on both quantitative and qualitative assays, the efficacy of acetosyringone was evaluated.…”

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confidence: 99%

Agrobacterium-mediated transformation of indica rice under Acetosyringone-free conditions

Rao

2007

Plant Biotechnology

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An optimized Agrobacterium -mediated transformation procedure for Phaseolus acutifolius A. Gray

Cited by 73 publications

References 49 publications

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

Biotechnology approaches to overcome biotic and abiotic stress constraints in legumes

Agrobacterium-mediated transformation of indica rice under Acetosyringone-free conditions

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