• Journal of Plant Biotechnology
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• 제36권2호
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• pp.137-143
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• 2009

An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

• Journal of Plant Biotechnology
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• 제36권4호
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• pp.373-383
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• 2009

To set efficient transformation system in chrysanthemum, thirty-four chrysanthemum (Dendranthema grandiflorum Kitamura) varieties were collected and cultured for shoot regeneration. Five varieties, ‘Shuho-no-chikara', ‘Zinba', ‘Baekma', ‘Pink pride' and ‘Keumsu' of them were selected, because they had a high shoot regeneration efficiency. MS medium containing 1.0 mg/L NAA and BA respectively was very adequate for shoot regeneration in those varieties. MS medium with 3.0 mg/L NAA and 1.0 mg/L kinetin in ‘Shuho-no-chikara' and the medium with 0.5 mg/L NAA and 3.0 mg/L BA in ‘Keumsu' were also suitable for shoot regeneration. The most efficient callus induction and shoot regeneration were obtained on MS medium. Shoot regeneration was enhanced more than 8% on MS medium with 0.3% phytagel and 10-15 mg/L putrescine. The best cultural material for shoot regeneration was stem. When stem was used as a culture material, shoot regeneration rate was increased more than 26% and the days to shoot regeneration was shortened about 14 days.

• 식물조직배양학회지
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• 제25권5호
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• pp.347-351
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• 1998

딸기 '수홍'의 기내배양된 잎과 탁엽 절편체로부터 기관형성을 통한 식물체 재생 방법을 확립하고자 실험을 수행하였다. 엽절편체 배양에서는 생장조절제 처리에 따라 NAA 첨가시에는 뿌리 재생, BA 첨가시에는 신초 재생이 이루어졌으며, BA 1.0㎎/L + NAA 0.2 ㎎/L 처리구에서 재생률 31.1%, 절편체당 신초수 1.7개로 가장 양호하였다. 탁엽절편체 배양에서는 생장조절제와 광조건의 처리를 실시하였는데, 명조건에서는 생장조절제 처리에 따른 뚜렷한 경향을 발견할 수 없었으나, 암조건에서는 고농도 BA와 저농도 NAA의 혼용처리가 신초 재생에 필요함을 알 수 있었다. 탁엽절편체는 BA 2.0 ㎎/L + NAA 0.1㎎/L 처리구에서 재생률 44.4%, 절편체당 신초수 4.0개로 가장 양호한 반응을 나타내었다. 재생된 신초는 NAA 0.1㎎/L 가 첨가된 MS배지에서 발근되었으며, 이어서 인공토양(vermiculite : perlite = 1 : 1)에서 성공적으로 활착을 유도할 수 있었다.

• Journal of Plant Biotechnology
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• 제46권2호
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• pp.114-118
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• 2019

Efficient protocol for plant shoot regeneration of Brassica juncea L. CZERN was established by using organic media components and growth stimulating factors of the vermicompost and coelomic fluids. Formulated organic plant tissue culture media (Vermicompost (30%) extracts supplemented with 20 mL/L coelomic fluid) have shown maximum shoot regeneration when compared with the Murashige and Skoog (MS) medium, which were supplemented with 1 mg/L 6-benzyladenine (BA) and 0.1 mg/L of Naphthaleneacetic acid (NAA). Cotyledon explants produced the highest shoot regeneration frequency from fourday-old germinated seedlings in comparison with non-germinated seedlings. The vermicompost extracts have proved to be the best organic plant growth media to induce shoots from cotyledons compared to the MS media. Statistically significant difference (P = 0.008) for the root length, shoot length (P=0.000350) and the leaves (P=0.375) of the mustard plantlets were analyzed successfully. The survival rate was 98% in the mustard cotyledons on the Vermicompost extract media and 63% on MS media respectively. The coelomic fluid also is much suitable to induce shoots from cotyledons at lower concentrations. It was also shown that the vermicompost extract, which comprised of humic acids along with coelomic fluid, affected shoot regeneration from the cotyledons. An efficient and organic shoot regeneration study was standardized and it can be applicable in the improvement of the economically important crops.

• 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.

• Plant Resources
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• 제2권1호
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• pp.10-15
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• 1999

In vitro shoot regeneration from cotyledon and hypocotyl explants derived from germinating mature Impatiens seeds. The induction of organogenetic tissue was also influenced by the cotyledon and hypocotyl. Multiple shoot induction was higher in hypocotyl than in the cotyledon explant with Thidiazuron and a NAA medium.

• Molecules and Cells
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• 제44권10호
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• pp.746-757
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• 2021

Plant somatic cells can be reprogrammed into a pluripotent cell mass, called callus, which can be subsequently used for de novo shoot regeneration through a two-step in vitro tissue culture method. MET1-dependent CG methylation has been implicated in plant regeneration in Arabidopsis, because the met1-3 mutant exhibits increased shoot regeneration compared with the wild-type. To understand the role of MET1 in de novo shoot regeneration, we compared the genome-wide DNA methylomes and transcriptomes of wildtype and met1-3 callus and leaf. The CG methylation patterns were largely unchanged during leaf-to-callus transition, suggesting that the altered regeneration phenotype of met1-3 was caused by the constitutively hypomethylated genes, independent of the tissue type. In particular, MET1-dependent CG methylation was observed at the blue light receptor genes, CRYPTOCHROME 1 (CRY1) and CRY2, which reduced their expression. Coexpression network analysis revealed that the CRY1 gene was closely linked to cytokinin signaling genes. Consistently, functional enrichment analysis of differentially expressed genes in met1-3 showed that gene ontology terms related to light and hormone signaling were overrepresented. Overall, our findings indicate that MET1-dependent repression of light and cytokinin signaling influences plant regeneration capacity and shoot identity establishment.

• 한국자원식물학회지
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• 제32권3호
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• pp.237-243
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• 2019

Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, 'Dawon', 'Pungsan', 'Daewon', 'Taekwang' and 'Chongdoo 1' by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, 'Dawon' and 'Pungsan'. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 mg/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in 'Dawon' cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

• Journal of Plant Biology
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• 제36권1호
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• pp.1-8
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• 1993

Polyamine contents and the activities of their main biosynthetic enzymes, arginine decarboxylase (ADC) and ornithine decarboxylase(ODC), were investigated in Populus leaf segments during adventitious shoot regeneration with the addition of 1 millimolar polyamine synthesis inhibitors. From the study of polyamine synthesis inhibitors, ODC was found to be the principal route of polyamine biosynthesis in adventitious shoot regeneration of Populus leaf segments. The ADC inhibitor difluoromethyl arginine(DFMA) and ODC inhibitor difluoromethyl ornithine(DFMO) strongly reduced the putrescine content. On the contarary, DCHA, an inhibitor of spermidine synthase, increased it. Spermidine content was decreased with the treatment of each polyamine synthesis inhibitor, but the inhibitory effect of DCHA was stronger than any other polyamine inhibitor. The decreased polyamine level by polyamine synthesis inhibitors was restored with the exogenously applied polyamine. Comparing the polyamine contents with the adventitious shoot regeneration rate, were observed a close correlation between spermidine content and adventitious shoot regeneration of Populus leaf segments.

• 한국작물학회지
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• 제49권1호
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• pp.52-60
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• 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.