• Plant Biotechnology Reports
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• 제4권2호
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• pp.139-148
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• 2010

Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

• Korean Journal of Plant Tissue Culture
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• 제24권6호
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• pp.319-322
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• 1997

The optimal conditions for callus formation and plant regeneration were determined by the manipulation of culture method in unpollinated ovary culture of rice. The effect of cold pretreatment on callus formation and plant regeneration varied with duration of pretreatment. The maximum frequency (38.7%) of plant regeneration was obtained from the unpollinated ovary pretreated for 10 days at $12^{\circ}C$. The ability of callus formation and plant regeneration was higher on the medium with picloram (1 mg/L) than that of 2, 4-D (1 mg/L). However, the high concentration of picloram increased markedly the frequency of albino plant from unpollinated ovary-derived callus. Floral parts inoculated as a unit play important roles in callus formation and plant regeneration. Best result was obtained when ovary with partial cut glume, pedicel and secondary branch as a unit was cultured.

• Journal of Life Science
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• 제17권6호통권86호
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• pp.822-824
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• 2007

The efficient system for plant regeneration from cotyledon and primary feat explants of lettuce was established. Plant regeneration efficiency was shown 91.3% from cotyledon and 85.9% from primary leaf explants of variety 'Jungtongpogi' in KN medium. Plant regeneration efficiency was also estimated with various plant regeneration media in variety' Chungchima', which was lowest plant regeneration efficient showing 35.4% from cotyledon and 30.3% from prima leaf explants in KN medium. Kl medium increased 77.9% and 80.7% of plant regeneration efficiencies from cotyledon and primary leaf explants of variety 'Jungtongpogi' were cultured on KN medium. In case of varie쇼 ‘Chungchima', efficient plant regeneration was shown when primary leaf explants were cultured on SH and KI media.

• Plant Resources
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• 제6권3호
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• pp.233-241
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• 2003

The shoot tip and young leaf of Rhodiola sachalinensis were cultured to invest the plant growth regulator condition for callus induction, shoot and root regeneration. When the shoot tip was sterilized in 2.0% of NaOCl for 20min., the contamination rate was the lowest. And the survival rate of the culture material was good in carbenicillin 500mg/L treatment group. Callus was obtained from shoot tip and young leaf segments. NAA 0.1-1.0mg/L and 2,4-D 0.1-0.5mg/L alone treatment were shown to have a good response on callus induction from shoot tip culture. In the case of young leaf culture, NAA and 2,4-D 0.1-0.5mg/L alone treatment were good in callus induction. In culturing shoot tip NAA 0.5mg/L and BA 0.5mg/L, NAA1.0mg/L and BA 0.lmg/L combination treatment was good in shoot regeneration. The regenerated shoots were rooted on MS medium supplemented with NAA and BA combination treatment. Especially, NAA 1.0mg/L and BA 0.1mg/L combination treatment was effective for root regeneration.

• Journal of Plant Biotechnology
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• 제7권2호
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• pp.129-134
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• 2005

Callus induction from a leaf explant has been achieved in Lilium Oriental hybrid 'Casa Blanca'. The highest frequency of callus induction was obtained on MS medium supplemented with 0.5 mg/L BA and 2.0 mg/L NAA after 2 months of culture. The cultures maintained continuously without change in color and type of callus when they cultured in the dark. Plantlet regeneration with a high frequency was achieved from induced calli on the same medium. A number of shoots are formed from one cluster of callus, and bulblets developed into intact plantlets after transfer to hormone-free MS medium. No phenotypic variations were observed among regenerants. Enhancement in plantlet regeneration via callus formation would be expected to facilitate the efficiency of transformation of this Oriental hybrid 'Casa Blanca'.

• Korean Journal of Medicinal Crop Science
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• 제14권2호
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• pp.87-91
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• 2006

An efficient plant regeneration of C. asiatica was achieved from organogenesis using petiole explants of in vitro plantlet on MS basal medium controled with different plant growth regulators (NAA,2,4-D, IAA kinetin, and BA). Best results that 50%, efficiency of regeneration per explant for regeneration were obtained with IAA $17.13\;{\mu}M$ and BA $8.9\;{\mu}M$. Formation of adventitious shoots via organogenesis from the petiole explant was verified by histological sectioning of plantlets. Regenerated plants were transplanted into soil.

• Journal of Plant Biotechnology
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• 제6권2호
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• pp.77-85
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• 2004

Development of efficient plant regeneration and genetic transformation protocols (using the Particle Inflow micro-projectile Gun and the shoot-tips as target tissue) of Sorghum bicolor (L.) Moench in terms of expression of the reporter gene, $\beta$-glucuronidase(uidA) is reported here. Two Indian cultivars of sorghum were used in the study, viz. M-35-1 and CSV-15. Plant regeneration was achieved from one-week-old seedling shoot-tip explants via multiple-shoot-clumps and also somatic embryos. The multiple-shoot-clumps were produced on MS medium containing BA (0.5, 1.0 or 2.0 mg/$L^{-1}$), with biweekly subculture. Somatic embryos were directly produced on the enlarged dome shaped expansive structures that developed from shoot-tip explants (without any callus formation) when cultured on MS medium supplemented both with BA (0.5, 1.0 or 2.0 mg/$L^{-1}$) and 2,4-D (0.5 mg/$L^{-1}$). Whereas each multiple-shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing 1.0mg/$L^{-1}$ IBA and successfully transplanted to the glasshouse and grown to maturity with a survival rate of 92%. The plant regeneration efficiency of both the genotypes were similar. After the micro-projectile bombardment, expression of uidA gene was determined by scoring blue transformed cell sectors in the bombarded tissue by an in situ enzyme assay. The optimal conditions comprising a helium pressure of 2200 K Pa, the target distance of 11 cm with helium inlet fully opened and the use of osmoticum have been defined to aid our future strategies of genetic engineering in sorghum with genes for tolerance to biotic and abiotic stresses.

• Journal of Plant Biotechnology
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• 제45권2호
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• pp.146-153
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• 2018

Lilium ledebourii Bioss is a wild species of Lilium, which grows naturally in some provinces of Iran. Previous studies on Lilium tissue culture have been linked to direct regeneration and a few studies have been conducted on indirect regeneration, which has been studied under bright conditions. In this study, for the first time in the world, all the stages of indirect regeneration (callus induction, shoot and root induction) have been studied under dark conditions. Callus formation and the regeneration levels of L. Ledebourii Bioss were examined for three replicates in an MS (Murashige and Skoog) medium with different hormonal compositions and by using a factorial experiment in the framework of a completely random plan. For callus initiation, 2,4-D and kinetin hormones were used in five and four levels, respectively, as auxin and cytokinin. Results showed that the highest percentage of the callus was found in $3{\mu}M$ of 2,4-D and $0.5{\mu}M$ of kinetin. In terms of callus wet weight, the highest amount was found in $3{\mu}M$ of 2,4-D and $0.5{\mu}M$ of kinetin. In addition, in terms of diameter, the highest amount was found in $3{\mu}M$ of 2,4-D, and $0.5{\mu}M$ of kinetin. In summary, the 2,4-D hormone had a major impact on the percentage of regeneration increase so that the best response was related to the composition of $3{\mu}M$ of 2,4-D, and $0.1{\mu}M$ of kinetin. This study contended that auxin and cytokinin can induce long shoots and roots through cell elongation in dark condition.

• Journal of The Korean Society of Grassland and Forage Science
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• 제24권3호
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• pp.265-270
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• 2004

Plant regeneration from seed-derived callus of Kentucky bluegrass(Poa pratensis L. cv. Kenblue) was investigated. Callus induced on the medium supplemented with 2 mg/L 2,4-D and 0.2 mg/L BAP showed highest frequency of plant regeneration on the regeneration medium supplemented with 1 mg/L NAA and 5 mg/L kinetin. Callus induced in the dark condition showed higher regenerability than that induced in the dim light. MS medium was better than N6 and B5 medium in enhancing plant regeneration. Maltose was superior to sucrose in plant regeneration as carbon source in the medium.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.241-246
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• 2005

Factors affecting regeneration of different Rubus varieties (blackberry, raspberry and their hybrid) were examined and a reliable regeneration system was established. Media for stock plant maintenance were tested; different explants and media were investigated to find the best circumstances for the regeneration. The effect of the commonly used antibiotics was studied to determine the most suitable one for selection of the transformants. We found that both MS and LS media supplemented by $20\;gL^{-1}$ sucrose are suitable for the stock plant maintenance. The optimal hormone content for the stock plants is $0.125\;mgL^{-1}$ 6-benzylaminopurine (BAP) with $0.01\;mgL^{-1}$ indole-3- butyric acid (IBA). The highest regeneration rate was observed on medium containing MS salts with B5 vitamins complemented with glucose, sucrose, maltose, $10\;gL^{-1}$ each, supplemented with benzylaminopurine riboside (BAR) ($2\;mgL^{-1}$) and indole-3-acetic acid (IAA) ($0.1\;mgL^{-1}$). The regenerated shoots appeared directly from the cut edges, without callus phase. Hygromycin and geneticin proved to be good selection agents for the Rubus explants, but due to their severe effect on the tissues we propose to use marker-free constructions for the transformation.