Producing Transgenic Thompson Seedless Grape (Vitis vinifera) Plants using Agrobacterium tumefaciens

Abstract

Under osmotic stress plant avoid it by different modification in their metabolisms such as increasing the synthesis of mRNA of delta 1-pyrroline-5-carboxylate synthase (P5CS), where P5CS is a target gene for increasing proline production and expect to improve the resistance to abiotic stress. To test this hypothesis, grape (Vitis vinifera L.) cv. Thompson Seedless was conducted to develop a protocol for high frequency regeneration and Agrobacterium mediated P5CS gene transfer. Callus induction was achieved by culture leaf explants on Nitsch and Nitsch (NN) basal medium including 2.0 mg L -1 2,4-di-chloro-phenoxy-aceticacid + 0.5 mg L -1 Thiadiazuron solidified with 2.5 g L -1 phytagel. While for in vitro proliferation of plant, the calli were cultured on NN medium supplemented with 3.0 mg L -1 Zeatin riboside + 0.5 mg L -1 Thiadiazuron. Agrobacterium-mediated transformation using the strain LB4404 harbouring the binary vector pBI121 with the P5CS gene under CaMV 35S promoter and the bar gene as a plant selectable marker was used for transforming grape explants. A P5CS specific band (2100 bp) was amplified from DNA extracted only from the transgenic grape plants and 24% for P5CS gene was positive for the bar gene. Over expression of the abiotic stress P5CS gene was enhanced synthesis of proline to over 6 times higher in transgenic plants compared to controls. The successful transformation of genetically diverse grape cv. Thompson Seedless indicated that the transformation system may have general application to an even wider range of grapes cultivars.

Genetic diversity assessment and in vitro propagation of some date palm (Phoenix dactylifera L.) varieties

“…Several studies have documented different interactions between date palm varieties and the growth regulators resulting in varied responses in the embryogenesis of calli (Eshraghi et al, 2005;Fki et al, 2003). Metwali et al (2020) found that the response to root formation and the lengths of shoots/roots of Magdoul variety improved by using IBA (0.5 mg L -1 ) in combination with NAA at 2.0 mg L -1 .…”

Prospek Dan Perkembangan Bioteknologi Mikroba

“…Pengkodean enzim phosphinothricin acetyltransferase (PAT) oleh gen bar memiliki kemampuan untuk mengubah phosphinothricin menjadi bentuk asetilasi nontoksik. Tanaman transgenik telah diperoleh dengan mentransfer gen tersebut ke banyak tanaman termasuk sorgum, kacang tunggak, kapas, kedelai, anggur, gula bit, aprikot, dan ubi jalar selama 30 tahun terakhir (Metwali et al, 2016).…”

Organogenesis in a broad spectrum of grape genotypes and Agrobacterium-mediated transformation of the Podarok Magaracha grapevine cultivar

“…Grapevine plants can also be regenerated via organogenesis (Thomas et al, 2000;Bettoni et al, 2015). For this purpose, fragments of shoot tips (Dutt et al 2007), fragments of internodes (Favre 1977; Rajasekaran and Mullins 1981; Kurmi et al 2011;Zhang et al, 2011), fragments of leaves (Clog et al, 1990;Stamp et al, 1990а,b;Torregrosa and Bouquet, 1996;Zhang et al, 2011;Nicholson et al, 2012;Ehab et al, 2016, Kumsa 2017) and fragments of petioles (Reisch et al, 1990;Li et al, 2007;Zhang et al, 2011) are often used as explants. However, reports on the use of direct organogenesis in genetic transformation from somatic tissues are extremely rare (Xie et al, 2016;Sabbadini et al 2019).…”