Agrobacterum-Mediated Transformation of Citrus

A genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an update

“…A. tumefaciens has been widely used for efficient delivery of viral genomes into different plants including a citrus-infecting virus and several phloem-limited viruses some of which belong to the Closteroviridae family (Grimsley et al, 1986; Prokhnevsky et al, 2002; Chiba et al, 2006; Vives et al, 2008; Liu et al, 2009; Wang et al, 2009). Although transient or stable transformation of citrus species has also been demonstrated (Cervera et al, 1998; Tzfira et al, 2004; reviewed in Peña et al, 2004a), previous trials to agroinfect citrus plants with CTV using different binary plasmids, silencing suppressors and disarmed Agrobacterium strains were unsuccessful (Gowda et al, 2005, and our unpublished data). Infiltration of citrus leaves with Agrobacterium is usually very inefficient for bacterial penetration and usually leads to low level of T-DNA expression, as observed with other recalcitrant species (Wroblewski et al, 2005).…”

Gene stacking in 1-year-cycling APETALA1 citrus plants for a rapid evaluation of transgenic traits in reproductive tissues

“…The 100 bp DNA ladder (Invitrogen) is included in each gel. (C) Southern blot analysis of retransformed AP1 plants was performed as described by Peña et al (2004). DNA was isolated from leaves according to Dellaporta et al (1983) and 20 g were digested with DraI or XhoI restriction enzymes.…”

“…Inoculation was carried out by immersion of explants for 15 min in a suspension of Agrobacterium in inoculation medium and cultured on CM for 3 days and then on SRM plus 500 mg/L cefotaxime, 250 mg/L vancomycin and assayed concentrations of selective agent. Transgenic plant recovery was achieved by in vitro grafting and subsequent greenhouse grafting on vigorous rootstocks (Peña et al, 2004). citrange tree becomes an excellent alternative to investigate flower and some fruit quality-related issues in citrus in a reasonably short period of time.…”

“…Here we demonstrate that by: (1) selecting as source material a single-copy AP1-transformant exhibiting consistent AP1 mRNA expression and conspicuous flowering every spring during at least five consecutive years, and (2) developing proper screening conditions to recover retransformants, the rapid-cycling Hygromycin and bialaphos regeneration tests. Forty Carrizo citrange epicotyl segments were cultured per treatment in cocultivation medium (CM) for 3 days, and then transferred to shoot regeneration medium (SRM) supplemented with 0, 2, 5, 10, or 20 mg/L of hygromycin (A) and 0, 0.5, 2, 5, 10, 20, or 50 mg/L of bialaphos (D) (Cervera et al, 1998;Peña et al, 2004). Shoot regeneration was examined 2-3 months after culture.…”

Transformação genética de tomateiro (<i>Solanum lycopersicum</i> cv. \'Micro-Tom\') e de laranja doce (<i>Citrus sinensis</i> L. Osbeck) com o gene <i>Csd</i>1 (superóxido dismutase do cobre e do zinco), isolado de<i> Poncirus trifoliata

“…The relation between the transgene transcript level with the resistance of plants to pathogens is set after inoculation with Candidadus Liberibacter. (SCHINOR, 2006;FEBRES et al, 2008;ZANEK et al, 2008;MUNIZ et al, 2012MUNIZ et al, , 2014, resistência a doenças causadas por fungos (PEÑA; NAVARRO, 1999;GENTILE et al, 2007;KATOH et al, 2007) e resistência a doenças causadas por bactérias (BOSCARIOL et al, 2006;PAOLI et al, 2007;MENDES et al, 2010;YANG et al, 2011).…”