Agrobacterium tumefaciens-mediated high frequency genetic transformation of an Indian cowpea (Vigna unguiculata L. Walp.) cultivar and transmission of transgenes into progeny

Chaudhury, Darshna; Madanpotra, Seema; Jaiwal, Ranjana; Saini, Raman Kumar; Kumar, Pramod; Jaiwal, Pawan K.

doi:10.1016/j.plantsci.2006.11.009

Cited by 71 publications

(52 citation statements)

References 26 publications

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“…The same pattern was confi rmed in all of the crosses between Olathe M 1/4 and the four commercial cultivars, suggesting that there was no recombination in the sequence of the transgene rep-trap-ren (Figure 1). Similar results were obtained by Chaudhury et al (2007), who registered the integration of two copies of the transgene in the genome of cowpea, where the transgene segregated in a 3:1 Mendelian pattern. In addition, Okada et al (2002) did not observe altered pattern of segregation, when they had multiple copies of the transgene introduced into the genome of sweet potato, also suggesting that the copies were inserted in a single locus.…”

Section: Resultssupporting

confidence: 85%

“…There are reports on the progeny of certain lines that inherited only one or none of the copies of the transgene. In many cases, the transgenes were inserted in different loci, resulting in the integration of multiple copies in the plant genome (Aragão et al, 1996;Rooke et al, 2003;Huang & Wei, 2005;Chaudhury et al, 2007). An insertional mutation of an essential gene required for ovule fertilization and development was indicated for aberrant inheritance of gus, neo, AC123 and BC1 transgenes in common beans (Aragão et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Transgene inheritances and genetic similarities of near isogenic lines of genetically modified common beans

Pinheiro

Faria

Nogueira

et al. 2009

Pesq. agropec. bras.

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-The objective of the present work was to determine the inheritance and stability of transgenes of a transgenic bean line expressing the genes rep-trap-ren from Bean golden mosaic virus and the bar gene. Crosses were done between the transgenic line and four commercial bean cultivars, followed by four backcrosses to the commercial cultivars. Progenies from each cross were evaluated for the presence of the transgenes by brushing the leaves with glufosinate ammonium and by polymerase chain reaction using specifi c oligonucleotides. Advanced generations were rub-inoculated with an isolate of Bean common mosaic necrosis virus (BCMNV). The transgenes were inherited consistently in a Mendelian pattern in the four crosses studied. The analyzed lines recovered close to 80% of the characteristics of the recurrent parent, as determined by the random amplifi ed DNA markers used, besides maintaining important traits such as resistance to BCMNV. The presence of the transgene did not cause any detectable undesirable effect in the evaluated progenies.Index terms: Phaseolus, BCMNV, genetic engineering, genetic variability, transgene genotypic stability. Herança de transgenes e similaridade genética de linhagens quase isogênicas de feijoeiro-comum geneticamente modifi cadoResumo -O objetivo do presente trabalho foi determinar a herança e a estabilidade de transgenes de uma linhagem de feijoeiro-comum com expressão dos genes rep-trap-ren, do Bean golden mosaic virus, e do gene bar. Foram realizados cruzamentos entre a linhagem transgênica e quatro cultivares comerciais de feijão, seguidos de quatro retrocruzamentos. As progênies de cada cruzamento foram avaliadas quanto à presença dos transgenes, com aplicação do glifosinato de amônia nas folhas e por meio da reação da polimerase em cadeia com uso de oligonucleotídeos específi cos. O vírus do mosaico comum necrótico do feijoeiro, Bean common mosaic necrosis virus (BCMNV), foi inoculado mecanicamente nas gerações avançadas. Os transgenes foram herdados em padrão mendeliano nos quatro cruzamentos estudados. As linhagens analisadas apresentaram cerca de 80% das características do parental recorrente, conforme determinado por análises com uso de marcadores de DNA, além de manter caracteres importantes, tais como resistência ao BCMNV. A presença do transgene não causou efeitos indesejáveis que pudessem ser detectados nas progênies avaliadas.Termos para indexação: Phaseolus, BCMNV, engenharia genética, variabilidade genética, estabilidade genotípica do transgene.

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Section: Resultssupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Transgene inheritances and genetic similarities of near isogenic lines of genetically modified common beans

Pinheiro

Faria

Nogueira

et al. 2009

Pesq. agropec. bras.

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“…Most of the Vigna species, Vigna unguiculata (Chaudhary et al 2007) and Vigna radiata (Jaiwal et al 2001;Sonia et al 2007) required 2-3 days cocultivation period for the maximum transformation frequency.…”

Section: Optimization Of Transformation Protocolmentioning

confidence: 99%

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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“…Although relatively few transgenes were confirmed in explants (17 of 161, or 10.5%), this ratio of transformation is higher than that which is commonly observed using the Agrobacterium system and in transformation rates reported for Begonia x hiemalis Fotsch (0-3%) (Kishimoto et al, 2002); Hordeum vulgare (0-7%) (Shrawat et al, 2007), and an Vigna unguiculata (1%) (Chaudhury et al, 2007). However, considering the influence of ultrasound as well as time of contact with the bacteria, there was a significant increase in the number of transformed plants.…”

Section: Discussionmentioning

confidence: 53%

Improving plant transformation using Agrobacterium tumefaciens

Neto¹,

Oliveira²,

Lourenço³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Here, we report a quick and low-cost method to improve plant transformation using Agrobacterium tumefaciens. This method involves the use of physical wounding, ultrasound, and an increase in exposure time to the bacteria. We show how the transformation rate increased from 0 to 14% when an ultrasound pulse of 10 s was used in conjunction with 96 h of bacterial exposure in Eclipta alba explants.

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Agrobacterium tumefaciens-mediated high frequency genetic transformation of an Indian cowpea (Vigna unguiculata L. Walp.) cultivar and transmission of transgenes into progeny

Cited by 71 publications

References 26 publications

Transgene inheritances and genetic similarities of near isogenic lines of genetically modified common beans

Transgene inheritances and genetic similarities of near isogenic lines of genetically modified common beans

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

Improving plant transformation using Agrobacterium tumefaciens

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