Agrobacterium tumefaciens-Mediated genetic transformation in tea (Camellia sinensis [L.] O. Kuntze)

Lopez, S. Joseph; Kumar, R. Raj; Pius, P.K.; Muraleedharan, N.

doi:10.1007/bf02772730

Cited by 36 publications

(21 citation statements)

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“…The advantages are (1) stable gene expression because of the insertion of the foreign gene into the host plant chromosome; (2) low copy number of the transgene; and (3) large size DNA segments can be transferred. Many crops, flowers and trees have been genetically modified using this method that are important from agronomic and horticultural point of view (Ko and Korban, 2004;Lopez et al 2004). The various Agrobacterium strains have been used to accomplish the tasks.…”

Section: Resultsmentioning

confidence: 99%

Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Bakhsh¹,

Anayol²,

Özcan³

2014

Emir. J. Food Agric

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Agrobacterium mediated transformation has been widely used for research in plant molecular biology and for genetic improvement of crops exploiting its tremendous ability to transfer foreign DNA to plants. In this study, the transformation efficiency of five Agrobacterium tumefaciens strains GV2260, LBA4404, AGL1, EHA105, and C58C1 was evaluated in Nicotiana tabacum L. cultivar Samsun. The Agrobacterium strains contained the recombinant binary vector pBin19 harboring beta-glucuronidase uidA gene under 35S promoter. Neomycin phosphotransferase (nptII) gene was used as a selectable marker at a concentration of 100 mg L-1 kanamycin. The expression of uidA gene in regenerated T0 plants was firstly analyzed by GUS histochemical analyses and later on confirmation of presence of the nptII and uidA genes in regenerated plants was determined by PCR. The highest transformation rate (20%) was obtained with the Agrobacterium strain LBA4404, followed by EHA105, GV2260, C58C1 and AGL1. The higher transformation efficiency achieved in our studies make LBA4404 Agrobacterium strain optimal for functional genomics and biotechnological applications in tobacco plants.

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Section: Resultsmentioning

confidence: 99%

Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Bakhsh¹,

Anayol²,

Özcan³

2014

Emir. J. Food Agric

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“…Further, the RNAi based on hpRNA strategy has been reported as more efficient for gene silencing (Smith et al 2000). Also methods are available for transformation and regeneration in tea (Lopez et al 2004;Bhattacharya et al 2006;Sandal et al 2007). Our recent study has documented the regulatory role of caffeine synthase (CS) in caffeine biosynthesis of tea (Mohanpuria et al 2009).…”

Section: Discussionmentioning

confidence: 97%

“…It was used successfully in the production of first healthy transgenic tea plants via Agrobacterium-mediated transformation (Mondal et al 2001a). Further, tea transformations and production of transgenics have been attempted via Agrobacterium or biolistic mediated approach by several workers using different explants such as embryogenic tissues, somatic embryos, and in vitro leaves (Lopez et al 2004;Jeyaramraja and Meenakshi 2005;Bhattacharya et al 2006;Sandal et al 2007). …”

Section: Paraxanthinementioning

confidence: 99%

Producing low-caffeine tea through post-transcriptional silencing of caffeine synthase mRNA

et al. 2011

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In this study, attempt has been made to produce a selected cultivar of tea with low-caffeine content using RNAi technology. The caffeine biosynthetic pathway in tea has been proposed to involve three N-methyltransferases such as xanthosine methyltransferase, 7-N-methylxanthine methyltransferase and 3, 7-dimethylxanthine methyltransferase. Last two steps of caffeine biosynthesis in tea have been known to be catalyzed by a bifunctional enzyme known as caffeine synthase. To suppress the caffeine synthesis in the selected tea [Camellia sinensis (L.) O. Kuntze] cv. Kangra jat, we isolated a partial fragment of caffeine synthase (CS) from the same cultivar and used to design RNAi construct (pFGC1008-CS). Somatic embryos were transformed with the developed construct using biolistic method. Transformed somatic embryos showed reduction in the levels of CS transcript expression as well as in caffeine content. Plants were regenerated from the transformed somatic embryos. Transgenic plants showed a significant suppression of CS transcript expression and also showed a reduction of 44-61% in caffeine and 46-67% in theobromine contents as compared to the controls. These results suggest that the RNAi construct developed here using a single partial fragment of CS gene reduced the expression of the targeted endogenous gene significantly. However, the reduction in theobromine content in addition to caffeine documented the involvement of this single CS in the catalysis of last two methyl transfer steps in caffeine biosynthesis of tea.

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“…Many of the woody species have appeared to be recalcitrant to genetic manipulation. Despite this, progress has been made in a number of species, for example, some of the Populus hybrids (Martín-Trillo and Martínez-Zapater 2002), sour orange (Ghorbel et al 2000), sweet orange (Cervera et al 2005), tea (Lopez et al 2004), water melon (Cho et al 2008), tomato and potato (Bříza et al 2008). In addition, attempts have been made to produce insect-resistant transgenic plants through the introduction and expression of foreign genes encoding insecticidal or insect-inhibiting proteins, such as Bt endotoxins, protease inhibitors and lectins (Zhou et al 1998, Kota et al 1999, Corbin et al 2001, Guo et al 2004, Jube and Borthakur 2007.…”

Section: ⎯⎯⎯⎯mentioning

confidence: 97%

Production of transgenic Pinus armandii plants harbouring btCryIII(A) gene

Liu¹,

Liu²,

Yang³

et al. 2010

Biol. plant.

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A synthetic chimeric gene SbtCryIII(A) encoding the insecticidal protein btCryⅢ(A), was transformed into Pinus armandii embryos and embryogenic calli using Agrobacterium tumefaciens. Polymerase chain reaction and genomic DNA Southern blot analysis showed that the SbtCryIII(A) gene was integrated into the genome of transgenic Pinus armandii plants, and Northern blot analysis indicated that the SbtCryIII(A) gene was transcribed.

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Agrobacterium tumefaciens-Mediated genetic transformation in tea (Camellia sinensis [L.] O. Kuntze)

Cited by 36 publications

References 0 publications

Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Producing low-caffeine tea through post-transcriptional silencing of caffeine synthase mRNA

Production of transgenic Pinus armandii plants harbouring btCryIII(A) gene

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