• Journal of Plant Biology
• /
• v.36 no.4
• /
• pp.377-382
• /
• 1993

Rice (Oryza saliva L.) calli containing both embryogenic callus (EC) and non embryogenic callus (NEC) regions were initiated from the mature seed on MS medium supplemented with 2.0 mg/L 2,4-D, 0.5 mg/L kinetin. The calli were developed into two callus type which can be distinguished by visual examination depending on color and appearance. In order to illucidate the polypeptide composition between EC and NEC, the total protein extracted from two types of callus was analysed by electrophoresis. By one-dimesional anlaysis of SDS-PAGE and Isoelectric focusing, several protein bands showed quantitative and qualitative difference in each type of callus. The further analysis of the total protein with two-dimensional electrophoresis showed at least 20 EC specific protein and 10 NE specific protein. Also 3 specific protein spots showing micro heterogeneity of 90, 65, 50 kD were detected in EC, while a series of acidic heterologous protein spots were visualized in NEC.in NEC.

• Journal of Plant Biotechnology
• /
• v.30 no.4
• /
• pp.369-373
• /
• 2003

In spite of potential benefits of anther culture, low productivity of plant regeneration in some genotypes; e.g. tonsil and indica rice, is one of the major obstacles for practical use of anther culture. This study was conducted to improve cold shock method and carbohydrate source for increasing the efficiency of anther culture in rice. The most common carbon source, sucrose was replaced to maltose, which has two molecules of glucose. Maltose increased callus induction 1.4-to 1.8-fold higher in japonica rice, 3.2-to 11.6-fold in tongil types and 2.7-fold in indica rice IR50. Callus induction was increased from 0.2％ to 12.5％ in maltose medium compared to the medium supplemented with sucrose plus glucose in indica rice "Tetep". A simple procedure of vacuum packaging of panicles during cold shock treatment prolonged not only anther viability more than 15 days but also increased callus induction more than 2-fold compared to open-air storage (conventional method). Combining of above two methods, callus induction was increased 28 to 56％ in japonica, 13 to 33％ in tonsil type and 12 to 31％ in indica rice. Plant regeneration was increased 14 to 35％ in japonica, 10 to 20％ in tonsil and 4 to 15％ in indica rice, respectively.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.30 no.3
• /
• pp.271-276
• /
• 1985

Effects of media and its components on callus induction and plant regeneration were studied to increase the cultural efficiency in rice anther culture. The N$_{6}$ basic medium gave better results in callus induction than those of MS or Miller. The medium used for callus induction affects the plant regeneration. The frequency of plant regeneration from callus grown on Miller basic medium was lower than those of N$_{6}$ or MS. Most of calli derived from anthers, above 90%, were induced from 20 days to 40 days after anther inoculation. The cultural efficiency of modified N$_{6}$ basic medium which was composed of 31.5mM KNO$_3$ and 1.75mM(NH$_4$)$_2$SO$_4$ as nitrogen sources was higher than those of N$_{6}$ basic medium. Combination of NAA and Kinetin showed better results than that of 2, 4-D only in cultural efficiency. Effect of DL-alanine on callus induction in Indica variety, IR40, showed better response in the anthers pretreated for 6 days at 12$^{\circ}C$.

• Korean Journal of Breeding Science
• /
• v.49 no.3
• /
• pp.170-179
• /
• 2017

Plant molecular farming has attracted a lot of attention lately in the field of mass production of industrially valuable materials by extending application of the plant as a kind of factory concept. Among them, protein expression system using rice(Oryza sativa L.) callus is a technology capable of mass culture and industrialization because of a high expression rate of a target protein. This study was carried out to develop an Agrobacterium-mediated transformation system to increase the utilization of rice callus. The transformation efficiency was improved by using the hand when seeds were de-husked for callus induction. Furthermore, we were possible induction of callus from 6 years old seed smoothly. Selection of the callus contained the target gene was required a cultivation period of at least 3 weeks, and the most efficient selection period was after 6 weeks of culture including one passage. This selection was confirmed that the gene was stably inserted into the genomic DNA of the plant cell by the southern blot analysis and progeny test. Such an efficient selection system of rice callus that can be cultured in the long term will be contribute to the industrialization of useful recombinant proteins using rice.

• Journal of Plant Biology
• /
• v.13 no.3
• /
• pp.17-19
• /
• 1970

Histological observation of micropore-originated haploid rice callus was reported previously. Present study was attempted to clarify the growth or development of the calli when they were transferred to differentiation media prepared exclusively for differentiation of plantlets. When the callus was transferred to differentiation medium, the cells and tissues became radially elongated. Meristematic tissues were present but few in number, and their structures were quite different from those grown in the propagaton medium. Differentiation of tracheid, chloroplast, and epidermis-like cell layer, and formation of gap in the callus tissue were more conspicuous in differentiation media. Approximately ten days after transfer of callus to differentiation medium, plantlet was formed.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.49 no.1
• /
• pp.52-60
• /
• 2004

The optimized in vitro culture system was investigated for improvement of regeneration efficiencies by observing the responses of scutella-derived callus of Korean rice (Oryza sativa L.). Large variations of callus induction (43.9-93.9%) and shoot regeneration (0-88.7%) were observed among the rice cultivars depending on medium. However, shoot regeneration was significantly improved by selected utilization of basal medium, growth regulators, and carbon sources. N6 basal medium was more efficient for embryogenic callus induction than MS or LS basal medium, while MS was superior to N6 for shoot regeneration. The calli of highly regenerative cultivars grew faster and showed higher rates of green tissue formation (GT) and shoot regeneration (SR) and lower rate of callus browning (CB) than those of recalcitrant cultivars. Although a higher level of kinetin stimulated the GT and SR in highly regenerative cultivars, $10\textrm{mgL}^{-1}$ kinetin generally suppressed the GT and SR, while CB was accelerated compared to $2\textrm{mgL}^{-1}$ kinetin. Additional benefits of sorbitol combined with maltose (or sucrose) under $5\textrm{mgL}^{-1}$ kinetin were certainly confirmed on regeneration efficiencies compared to sucrose alone as carbon source and osmotic regulator. This combination showed high rate of GT and SR with multiple shoots while low rate of CB. With MSRK5SM-Pr medium ($5\textrm{mgL}^{-1}$ kinetin, 3% sorbitol, 2% maltose, $500\textrm{mgL}^{-1}$ proline), the regeneration efficiencies of total 17 out of 24 cultivars were practically improved 160% on average compared to MSRK2S ($2\textrm{mgL}^{-1}$ kinetin, 3% sucrose) control medium. Especially, the medium was most effective to the cultivars showing a medium level of regenerability such as Daesanbyeo and Dongjinbyeo and Suwon477, enhancing efficiencies more than 300-600% compared to MSRK2S medium.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.41 no.6
• /
• pp.729-735
• /
• 1996

This study was carried out to investigate the effect of Germaniwn(Ge) treatment in the culture media on the Ge absorption by the callus induced from brown rice cv. Dongjinbyeo. MS medium was more effective on the growth ratio of callus and the content of Ge and some inorganic elements except K in callus than N$_{6}$ medium. The more Ge treatment in the N$_{6}$ or MS medium, the more Ge absorption by the callus, but the growth ratio of callus and the content of Ca, Mg, and K in callus were decreased. The content of Fe, Mn, Zn, and Cu was increased under treatment up to 100～200mg /$\ell$ Ge, but tended to be decreased under treatment more than that of Ge concentration. Under treatment less than 150mg /$\ell$ Ge, GeO$_2$(inorganic Ge) was more effective on the Ge absorption by callus than Ge-132(organic Ge), but Ge-132 was more effective on the Ge ab-sorption by callus and the activity of callus in case of treatment more than 150mg /$\ell$ Ge. The lower pH of culture medium, the higher Ge content in the callus. When callus was cultured on medium supplemented with Ge and 0.1～1.0mM of citric acid or myo-inositol, content of Ge and some inorganic elements in callus, as well as growth and dry weight of callus, were tend to increase in comparison to control, but myo-inositol was more effective on them than citric acid.cid.

• Korean Journal of Plant Tissue Culture
• /
• v.27 no.2
• /
• pp.83-87
• /
• 2000

To investigate the effect of growth regulators and antioxidant in anther floating cultures of rice, anthers were cultured in liquid media supplemented with different growth regulators, and the effect of antioxidant mixture : (Sigma Chemical Co.) on callus induction and plant regeneration were investigated. N6 medium with 1 mg/L 2,4-D and 1 mg/L kinetin was the best for rice anther floating cultures, which showed 48.5% of callus induction and 6.8% of green plant regeneration. The callus induction was not affected by antioxidant mixture in liquid medium, and antioxidant mixture (250 mg/L) was effective for the reduction of brownish callus and improvement of plant regeneration Antioxidant mixture showed better effectiveness when it was supplemented to both media for callus induction and plant regeneration.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.29 no.3
• /
• pp.232-241
• /
• 1984

A series of experiments were carried out to know the effects of pollen stage, cold shock temperature and duration, and media for callus and green plant induction in rice anther culture. The results indicated that: (a) uninucleate stage of pollen was the most suitable stage for effective callus induction, (b) cold shock temperature of 8$^{\circ}C$ and 12$^{\circ}C$ was appeared to be proper temperature for callus induction, (c) callus induction rate was increased in the eight to 12 days long cold storage, (d) the medium N6 was better than that of N6D for callus induction, (e) green plant induction was better in both 4$^{\circ}C$ and 8$^{\circ}C$ than that of 12$^{\circ}C$ cold shock, (f) green plant frequency was higher in eight to 12 days long cold storage and (g) green plant frequency was doubled in the MS medium when compared with N6 medium.

• Journal of Plant Biotechnology
• /
• v.4 no.1
• /
• pp.39-44
• /
• 2002

The fast-migrating cationic peroxidase isozyme, named RC3, was purified from rice (Oryza sativa cv. Nak-Dong) callus. Purification of the enzyme was accomplished by ammonium sulfate fractionation, CM-cellulose ionexchange chromatography, and Sephacryl S-100 gel filtration. The molecular mass of the enzyme was about 34 KDa as determined by SDS-PACE and 38 KDa by Sephacryl-100 gel filtration. The pI value of the enzyme was 8.9. Antiserum against RC3 was raised in rabbits, and anti RC3 antiserum reacted with RC3 isozyme by Ouchterlony double immunodiffusion. The optimum pHs and Km values of the enzyme for various substrates were determined. Kinetic studies with various substrates showed that RC3 had very low Km value of 0.01 mM for ferulic acid and ascorbic acid. However, the enzyme did not use esculetin as a substrate.