The Establishment of in vitro Culture and Genetic Transformation of the Wheat Glu-1Dx5 Gene to Rice Plants by Gene Gun Bombardment （イネの培養系確立と遺伝子銃によるコムギGlu-1Dx5遺伝子の導入）
1.Identification of Callus Induction Potential of 15 Indonesian Rice Genotypes.
Nono Carsono and Tomohiko Yoshida. PPS 9(1):65-70(2006)
Abstract : The callus induction potential of 15 indica rice genotypes from Indonesia was examined in comparison with that of the japonica rice Nipponbare. Callus was induced from embryos of mature seeds (seed explants) and root segments (root explants) on MS and CI media. There was genotype x medium x explant interaction for inducing calli with a white/yellow structure and for browning, but not for callus induction ability and average diameter of callus. Four indica types, Fatmawati, Ciapus, BP-23 and BP-360-3, had callus-related traits similar to those of Nipponbare. Culture of seed explants on MS was more suitable for callus induction than that of root explants and these explants on CI medium, respectively.
2.Plant Regeneration Capacity of Calluses Derived from Mature Seed of Five Indonesian Rice Genotypes. Nono Carsono and Tomohiko Yoshida. PPS 9(1):71-77(2006)
Abstract : Establishment of a suitable plant regeneration system from mature seed derived callus of five rice genotypes is a prerequisite for genetic transformation. Selecting the most suitable medium and assessing the genotype performance for callus proliferation and regeneration are essential requirement for that purpose. The result showed that callus proliferation was affected by: genotype for the size of cluster of calli (SCC) and medium for calli proliferation capacity as well as SCC. Plant regeneration capacity was affected by the interaction effect between genotype and medium referring to green shot bud induction frequency and calli regenerable capacity. Plant regeneration capacity was affected independently by genotype and medium. Culture media D1 and NB5 were identified as the most suitable media for subculture and regeneration, respectively. Genotypes Fatmawati and BP-140 consistently performed best at the subculture and regeneration.
3.Evaluation of Spikelet-related Traits of Rice Plants Regenerated from Mature Seed-Derived Callus Culture. Nono Carsono1 and Tomohiko Yoshida PPS 10(1):86-90(2007.1).
Abstract Callus is an excellent source for in vitro plant regeneration, but plants regenerated from callus sometimes show phenotypic and genotypic variation from the initial plant. A phenotypic evaluation on spikelet-related traits of the rice plants regenerated from calluses was performed to assess the feasibility of callus culture in regenerating fertile plants. Results revealed that the phenotypic variation in spikelet-related traits of the regenerated plants was not always in a negative direction (reduction in their mean value). For instance, panicle length, spikelet number and fertile spikelet number of Ciapus and BP-23 in the regenerated plants were significantly greater than those of the initial plants (developed from seed). The spikelet fertility of the regenerated-plants was not significantly different from that of the initial plants except in Ciapus and BP-23. Results suggested that callus culture used in this study is appropriate for regenerating fertile rice plants and that some regenerated variants can be conveniently exploited for the breeding programs.
4.Transient Expression of Green Fluorescent Protein Gene in Rice Calluses:Optimization of Parameters for Helios Gene Gun Device
Nono Carsono and Tomohiko Yoshida. PPS 11(1):88-95,2008.
Abstract : An optimized condition for particle bombardment is necessary for efficient genetic transformation. In the present study, parameters for Helios gene gun, another device for cell transformation, were optimized based on transient synthetic green fluorescent protein [sGFP (S65T)] gene expression in rice calluses of indica Fatmawati and japonica Nipponbare. Parameters that found to be the most favorable conditions for transient expression of sGFP gene in rice callus cells were as follows: 250 psi helium pressure, 0.6 m gold particle size, 0.25 mg gold particles per shot, and 1.5 g plasmid-DNA per shot. Desiccation of callus cells for eight min. was also found appropriate. The level of transient sGFP gene expression was not significantly influenced by the pre-culture for 4 to 12 days before bombardment or by callus age between 10 and 33-weeks old in Fatmawati. Optimized parameters for this particular device should improve the transient expression, thus enabling stable expression of inserted genes via Helios gene gun using callus as a target tissue.
5. Genetic Transformation of the High Molecular Weight Glutenin (Glu-1DX5)
to Rice cv. Fatmawati
Yoshiharu Wada, Nono Carsono, Anas, Ly Tong and Tomohiko Yoshida.
Plant Prod. Sci. 12(3): 341-345 (2009.7).
Glu-1Dx5 gene encoding a high molecular weight (HMW) glutenin subunit Dx5
was co-transformed to rice callus cells of cv. Fatmawati. T1 and T2 plants revealed
that some transgenic plants carried the Glu-1Dx5 gene. Analysis of the endosperm
extracts of T2 plants by SDS-PAGE revealed the existence of a protein similar in
size to the wheat Glu-1Dx5 gene product.
6. コムギグルテニン遺伝子Glu-1Dx5 を導入したイネ玄米全粒粉の製パン特性
和田義春・氏家綾子・Nono Carsono・吉田智彦．日作紀 in press．
September 2001-January 2003: MSc in Plant Breeding, Wageningen University & Research Centre, The Netherlands
September 1991-August 1996: BSc in Plant Breeding, Faculty of Agriculture, Padjadjaran University, Indonesia
August 1997-August 2001: Basic Genetics, Advanced Genetics, Introduction to Plant Breeding
February 2003-present: Basic Biotechnology, Introduction of Molecular and Cellular Biology