Transformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants

Abstract

Cotton (Gossypium hirsutum L.) cotyledon tissues have been efficiently transformed and plants have been regenerated. Cotyledon pieces from 12-day-old aseptically germinated seedlings were inoculated with Agrobacterium tumefaciens strains containing avirulent Ti (tumor-inducing) plasmids with a chimeric gene encoding kanamycin resistance. After three days cocultivation, the cotyledon pieces were placed on a callus initiation medium containing kanamycin for selection. High frequencies of transformed kanamycin-resistant calli were produced, more than 80% of which were induced to form somatic embryos. Somatic embryos were germinated, and plants were regenerated and transferred to soil. Transformation was confirmed by opine production, kanamycin resistance, immunoassay, and DNA blot hybridization. This process for producing transgenic cotton plants facilitates transfer of genes of economic importance to cotton.

Transformation of Virus-Resistant Genotype of Gossypium hirsutum L. with Pesticidal Gene.

“…In present study, combination of biolistic and Agrobacterium was used for the transformation studies. The transformation system presented here differs in many ways from the work previously reported by Firoozabady et al, (1987) and Umbeck et al, (1987). In the present study, mature embryos was used as explant for the transformation of cotton.…”

“…In the present study, mature embryos was used as explant for the transformation of cotton. Previously callus and shoot apices were reported to be used as an explant for the transformation of cotton (Chlan et al, 1995;Firoozabady, 1987;Umbeck et al, 1987). The main advantage of mature embryo transformation is the plant development within two months in case of recalcitrant varieties.…”

“…Agrobacterium -mediated transformation is most commonly used method for cotton transformation. But difficulties exist due to low efficiency of regeneration of cotton plants and low efficiency of transformation (Firoozabady et al, 1987;Grierson and Covey, 1988). To overcome this barrier of regeneration, particle bombardment method has been used to transform recalcitrant crops (Christou,1992;John and Keller, 1996;McCabe et al, 1988;McCabe and Martinell, 1993).…”

Phytochrome RNAi enhances major fibre quality and agronomic traits of the cotton Gossypium hirsutum L

“…Coker-312 is the only G. hirsutum line that is easily amenable to tissue culture regeneration 12,31,32 ; however, it is not a commercially competitive fibre producer. While our RNAi construct was initially introduced and tested in Coker-312, a critical challenge was to show genetic stability and transferability of this construct and its resulting suite of traits to elite commercial cultivars.…”

“…We used a combination of available methodologies for cotton tissue culture 12,31,32 . Hypocotyl sections (5-7 mm thickness) from 1-week-old seedlings of Coker-312 were wounded with a laboratory razor and placed on P1 medium (4.31 g l À 1 Murashige and Skoog basal salt mixture (MS salt), 0.4 mg l À 1 thiamine HCl, 100 mg l À 1 myoinositol, 0.75 mg l À 1 MgCl 2 , 3% glucose, 0.2% phytagel, 5 mg l À 1 N 6 -(2-isopentenyl) adenine, 0.1 mg l À 1 1-naphthaleneacetic acid, pH 5.8).…”