<i>Agrobacterium</i>-mediated Transformation of Javanica Rice cv. Rojolele

Rachmawati, Diah; Hosaka, Takehiko; Inoue, Eiichi; Anzai, Hiroyuki

doi:10.1271/bbb.68.1193

Cited by 36 publications

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“…In the present study, genotypic differences in callus induction, subculture, and plant regeneration of Javanica rice cultivars were examined using C-modified medium (Rachmawati et al 2004) and MS2 medium (Dong et al 1996) As results, significant difference was found in callus induction rate and quality of callus induced among the five genotypes of Javanica rice (Table 2). Although a considerable variation was observed, all genotypes of rice can induce callus on both media.…”

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confidence: 87%

“…The overall transformation efficiency of the five cultivars revealed that Rojolele rice cultivar has a high transformation efficiency (23%), similar to that of the D. Rachmawati and H. Anzai Japonica rice cv. Nipponbare (Rachmawati et al 2004). Whereas transformation efficiency of four other Javanica rice cultivars was in the range of 3 to 10% (Table 3).…”

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confidence: 98%

“…Additionally, the quality of calli might be a key factor for the success in transformation (Lin and Zhang 2005). The type-I calli derived from mature seeds of each cultivar were infected for 5 min with Agrobacterium tumefaciens strain EHA101 containing the binary vector pAFT14 for transformation as described previously (Rachmawati et al 2004). After 3 days of co-cultivation, the infected calli were washed with sterile water containing 250 mg l Ϫ1 carbenicillin and transferred to C-modified medium containing 250 mg l Ϫ1 carbenicillin and 50 mg l…”

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confidence: 99%

“…Callus induction is the first step in rice transformation to obtain embryogenic calli (Rachmawati et al 2004). In particular, genotype and the composition of culture medium are important factors for successful embryogenic callus induction and regeneration of plant.…”

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Studies on callus induction, plant regeneration and transformation of Javanica rice cultivars

Rachmawati

Anzai

2006

Plant Biotechnology

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The purpose of this study was to develop a reproducible efficient procedure for the transformation of Javanica rice cultivars from Indonesia. Five rice cultivars cultivated in Indonesia now were examined for their capacity on the callus growth, plant regeneration and transformation. Their potential was affected by genotype and medium. Regarding the quality of callus, type I calli produced higher plant regeneration frequency than type II calli. So type I calli were inoculated with A. tumefaciens harboring binary plasmid pAFT14, which had a hygromycin resistance (hpt) gene and a gus gene. In this study, we examined two media for two steps in the tissue culture process for transformation, i.e. callus inducing and plant regeneration. The results show that C-modified medium was the suitable media for callus induction in the most Javanica rice cultivars. Agrobacterium-mediated transformation system has been extended to five Javanica rice cultivars. Among them, Rojolele consistently gave the best performance.Key words: Callus, Javanica rice, transformation.Plant Biotechnology 23, 521-524 (2006) This article can be found at http://www.jspcmb.jp/ Tissue Culture Note tolerant to drought (gogo rice). The callus induction rate was calculated after 4 weeks of culture as the percentage of the seeds producing callus.As results, significant difference was found in callus induction rate and quality of callus induced among the five genotypes of Javanica rice (Table 2). Although a considerable variation was observed, all genotypes of rice can induce callus on both media. Two types of calli, embryogenic and non-embryogenic ones, were obtained from the embryo scutellum. The embryogenic calli (type-I) were relatively dry, milky white in color, compact and nodular in appearance ( Figure 1A, B). In contrast, non-embryogenic calli (type-II) were soft and watery, mostly white but some brown in color ( Figure 1C, D). Four weeks after induction, the calli were subcultured onto fresh medium, which had the same composition as that used for callus induction. Following the subculture, the type-I calli grew about 5 to 10-fold in volume 30 days after transfer. However, the type-II calli turned brown and died.In general, callus induction rate of the five Javanica rice cultivars on C-modified medium was higher than that on MS2 medium (Table 2). This finding is consistent with the results reported by Ogawa (2000) that Cmedium is applicable to a wider range of cultivars including Indica, Japonica and Javanica subspecies. The induction rate of type-I calli from all the five Javanica rice cultivars on C-modified medium was higher than that on MS2 medium, especially for Rojolele. On Cmodified medium, the induction rate of Rojolele was 10 times higher than one on MS2. On the other hand, the induction rate of type-II calli of the five cultivars on MS2 medium was almost the same as that on C-modified medium (Table 2). To determine the regeneration ability, the initiated calli were transferred to a regeneration medium which was composed of ma...

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confidence: 98%

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Studies on callus induction, plant regeneration and transformation of Javanica rice cultivars

Rachmawati

Anzai

2006

Plant Biotechnology

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“…Scutellum-derived calli from mature seeds were co-cultivated with A. tumefaciens strain EHA101 that carried either pIG200 or pIG211. Agrobacterium-mediated transformation was carried out as previously described (Rachmawati et al 2004). After co-cultivation, the infected calli were washed with sterile water containing 250 mg/l carbenicillin and transferred to a C-modified medium containing 250 mg/l carbenicillin and 50 mg/l hygromycin.…”

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Production and Characterization of Recombinant Human Lactoferrin in Transgenic Javanica Rice

et al. 2005

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The objective of the present study was to develop a functional crop that could contribute to the maintenance and improvement of human health by the introduction of the human lactoferrin (hLF) gene. Lactoferrin is an 80-kDa iron-binding glycoprotein that has been considered to play many biological roles, including the regulation of iron absorption, protection against microbial and virus infection, stimulation of the immune system and cellular growth promotion. We introduced two different constructs containing either the native signal peptide from human lactoferrin (pIG211) or the signal peptide from rice glutelin (pIG200) fused to mature human lactoferrin into Javanica rice cv. Rojolele by using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the maize ubiquitin-1 promoter was detected in all the tissues of the transgenic plants. We found that the transgenic rice plants IG200R produced a considerable amount of recombinant hLF (rhLF) in seeds, accounting for approximately 15% of the total soluble protein (TSP). RhLF was purified from mature seeds by cation-exchange chromatography. Amino acid sequencing confirmed that the N-terminal sequences of rhLF for both constructs were identical with those of native LF from human milk. Rice rhLF showed a slightly smaller molecular weight than the native hLF. Immunofluorescence microscopy revealed that rhLF was located in the intracellular and intercellular regions of endosperm cells. Protein extracts from transgenic rice seeds exhibited an antibacterial activity against Bacillus subtilis ATCC6633. The use of signal peptides and a constitutive ubiquitin-1 promoter for successful production of transgenic Javanica rice expressing a high level of rhLF was examined.

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Rice, Indica (Oryza sativa L.)

Hiei

Ishida

Komari

2014

Methods in Molecular Biology

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Indica varieties, which are generally recalcitrant to tissue culture and transformation, occupy 80 % of rice cultivation area in the world. Therefore, transformation method for indica rice must be improved greatly so that global rice production would take full advantage of cutting-edge biotechnology. An efficient protocol for indica transformation mediated by Agrobacterium tumefaciens is hereby described. Immature embryos collected from plants in a greenhouse are cocultivated with A. tumefaciens after pretreatment with heat and centrifuging. The protocol was successfully tested in many elite indica cultivars such as IR8, IR24, IR58025B, IR64, IR72, Suweon 258, and Nanjing 11, yielding between 5 and 15 of independent transgenic plants per immature embryo. The use of immature embryos is recommended because gene transfer to them could be much more efficient and much less genotype dependent than gene transfer to callus.

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Agrobacterium-mediated Transformation of Javanica Rice cv. Rojolele

Cited by 36 publications

References 19 publications

Studies on callus induction, plant regeneration and transformation of Javanica rice cultivars

Studies on callus induction, plant regeneration and transformation of Javanica rice cultivars

Production and Characterization of Recombinant Human Lactoferrin in Transgenic Javanica Rice

Rice, Indica (Oryza sativa L.)

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