Stable genetic transformation of castor (Ricinus communis L.) via particle gun-mediated gene transfer using embryo axes from mature seeds

Abstract

The first successful attempt to produce stably transformed castor plants through direct gene transfer using particle gun (BioRad) is described. Decotyledonated embryos from mature seeds were germinated and the embryonic axis was induced to proliferate on Murashige and Skoog (MS) medium supplemented with 0.5 mg l(-1) thidiazuron (TDZ) and subjected to bombardment after 5-7 days of pre-incubation. The physical parameters for transient transformation were optimized using the UidA gene encoding beta-glucuronidase (GUS) as the reporter gene and with hygromycin-phosphotransferase (hptII) gene as selectable marker. Statistical analysis revealed that helium pressure, target distance, osmoticum, microcarrier type and size, DNA quantity, explant type and number of bombardments had significant influence on transformation efficiency, while the effect of genotype was non-significant. Of the different variables evaluated, embryonic axes from mature seeds, a target distance of 6.0 cm, helium pressure of 1,100 psi, 0.6 microm gold microcarriers, single time bombardment and with both pre- and post-osmoticum were found ideal. Selection of putative transformants was done on MS medium supplemented with 0.5 mg l(-1) BA and hygromycin (20, 40 and 60 mg l(-1)) for 3 cycles. The stable integration of the incorporated gene into castor genome was confirmed with PCR and Southern analysis of T0 and T1 plants. Transformation frequency in terms of plants grown to maturity and showing the presence of the introduced genes was 1.4%. The present results demonstrate the possibility of transformation of embryonic meristematic tissues of castor through particle delivery system.

Apis mellifera pollination improves agronomic productivity of anemophilous castor bean (Ricinus communis)

“…Since ancient times it has been exploited for its oil which was used as ingredient to cosmetics, shampoo, soap, hand lotion, laxative, fuel for lamps and as a high speed lubricant (Capasso et al 1994, Copley et al 2005, Morris et al 2011. Nowadays, castor bean is cultivated mainly for biodiesel production because the oil content of its seeds reach up to 50% (Melo et al 2008, Sailaja et al 2008, Vanaja et al 2008.…”

Science Behind Cotton Transformation

“…In case of Ricinus (castor oil plant), transformation through gene gun is dependent on many factors such as helium pressure, target distance, osmoticum, microcarrier type and size, DNA quantity, explant type, and number of bombardments had significant influence on transformation efficiency (Figure 2). Recommended distance is 6.0 cm, helium pressure of 1100 psi, 0.6-μm gold microcarriers, furthermore single bombardment has shown positive results [46]. Finer and McMullen [47] demonstrated the successful recovery of transgenic cotton plants by biolistic bombardment of embryogenic suspension cultures and reported a transient to stable conversion frequency of about 0.7%.…”

Characterization Of Slovak Castor Genotypes (Ricinus Communis L.) Using Molecular Markers

“…Due to its unique chemical and physical properties, the oil from castor seed is used as raw material for numerous and varied industrial applications, such as: manufacture of polymers, coatings, lubricants for aircrafts, cosmetics, etc, and for the production of biodiesel (Jeong and Park, 2009). With more than 95% of the world's castor production concentrated in limited parts of India, China, and Brazil (Sailaja et al, 2008), and because of the ever increasing world-wide demand of castor for industrial use, there is a pressing need to increase the hectarage and productivity of castor. Castor is a cross pollinated crop and is usually cultivated as a hybrid in India, as hybrids give significantly greater yields than pure lines or varieties (Birchler et al, 2003;Reif et al, 2007).…”