• Journal of Plant Biology
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• 제38권2호
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• pp.143-151
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• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• Journal of Plant Biotechnology
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• 제42권3호
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• pp.228-234
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• 2015

본 실험은 희귀 및 멸종위기 식물인 미선나무의 효과적인 증식을 위하여 줄기 증식, 생장 및 발근에 미치는 절편의 위치 효과 및 치상 방법의 효과를 구명하기 위해 실시되었다. 줄기유도는 BA를 처리한 액아절편 배양이 효과적이었으며 절편 당 2.4개의 줄기가 유도되었다. 정아 및 액아절편을 BA 1.0 mg/L를 전처리하여 수평과 수직으로 치상하였을 때 줄기유도는 액아 절편의 수직치상으로 절편 당 2.5개로 가장 좋았다. 반면 줄기의 생장, 발근 및 뿌리 발달은 정아절편의 수직치상에서 양호하였고 BA의 전처리 시간이 길어질수록 발근이 억제되는 경향을 보였다. 기내증식된 줄기의 정아 및 액아를 절편으로 기외에서 미세삽목(micro-cutting)을 실시하여정아절편에서 100% 발근되었고, 뿌리수는 개체 당 평균 6.2개로 가장 양호하였다. 미세삽목묘는 온실에서 95% 이상 순화되었다. 이상의 결과로 볼 때 미선나무의 기내 증식은 액아를 절편으로 BA 처리 후 수직 치상하는 방법이 효과적이며, 줄기의 생장 및 발근은 정아를 절편으로 수직치상 하는 것이 효과적으로 나타났다. 또한 정아를 절편으로 미세삽목을 통해 효과적인 발근 유도 및 순화가 가능하여 미선나무의 실용적인 묘목생산이 가능함을 보여주었다.

• 대한치과교정학회지
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• 제26권6호
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• pp.667-676
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• 1996

We have examined the in vitro stage-related chondrogenic potential of human mandibular and limb bud mesenchyme cells using micromass culture. Our results indicate that limb bud mesenchyme cells as early as stage 16 by Carnegie system (37 days), well before the initiation of in vivo chondrogenesis, have chondrogenic potential which is expressed in micromass culture. These results are correlated with stage-related chondrogenic potential of human limb bud in vivo as a result of Alcian blue staining. The proliferation of chondrogenic cells increased in the first 3 days after culture and then decreased. These results were correlated with the cell cycle analysis of which the number of $G_0^1/G_1$ phase increased markedly after 3 days of culture, while the percentage of cells in S phase was decreased. On the other hand, it was rarely differentiated in the mandible. We examined the effects of two PKC modulators such as phorbol 12-myristate 13-acetate (PMA), a potent activator of PKC, and staurosporine (STSN), an inhibitor of PKC. PMA inhibited the chondrogenesis, whereas STSN promoted the chondrogenesis in a dose dependent manner. In addition, PMA exerted no inhibitory effect when the cells were pretreated for 24 h with STSN, implying that the chondrogenic events might be settled at an early step in vitro and FKC may act as a negative modulator. Collectively, these results demonstrate, for the first time, the stage-related chondrogenic potential of human mandibular and limb bud mesenchyme cells and the role of PKC during chondrogenesis in vitro & in vivo.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.15-19
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• 2006

An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

• 식물조직배양학회지
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• 제28권3호
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• pp.173-173
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• 2001

• 한국연초학회지
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• 제16권1호
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• pp.64-68
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• 1994

The present experiments were conducted to investigate some of the factors affecting the number of haploids derived from anther culture of Nicotiana tabacum. Anther - donor plants grown under controlled environment room at 3$0^{\circ}C$ yielded more haploid than room at 18, 25 and 26-22-18$^{\circ}C$ in anther culture. Donor plants starved of fertilizer yielded more haploids as compared to those of the well fed with fertilizer in anther culture. Pretreatment of exercised flower bud at 5$^{\circ}C$ was shown to be more effective in anther culture than pretreatment at 7 and 1$0^{\circ}C$, and the optimum temperature and period of pretreatment were 4 or 6 days at 5$^{\circ}C$.

• Journal of Plant Biotechnology
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• 제6권2호
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• pp.103-106
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• 2004

Plantlet regeneration from the shoot apex was studied in three different genotypes of the chinese yam (Dioscorea opposita Thunb) cv. Jnagma and Danma, Dunggunma. The effects of plant growth regulators and inorganic salts concentration of the culture medium on bud induction and shoot growth were examined. The combinations of 0.2 mg/L BAP + 0.2mg/L kinetin, 0.01mg/L NAA + 0.2 mg/L kinetin and a single treatment of 0.2mg/L BAP were equally effective for bud and shoot formation from the shoot apices in the three cultivars. Auxin (2,4-D, NAA) treatment enhanced calli formation from the cultured apices. Also, the shoot apices of the cv. Dunggunma produced more callus and buds on the culture medium (MS) containing 0.05mg/L NAA and 0.5-1.0mg/L SAP. Lower salt strength of medium inhibited shoot elongation but did not have much effect on the shoot and bud induction from the shoot apices. These results will be useful to obtain disease-free plants of the Chinese yam.

• 원예과학기술지
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• 제33권1호
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• pp.18-23
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• 2015

Time-specific responses of flower bud differentiation were investigated in 'Shinko' (Pyrus pyrifolia Nakai) pear grown at different altitudes from July through December 2013 to determine their suitability as scions in a top-grafting system. Flower bud initiation and bud necrosis were monitored on each of three sections of one-year-old shoots: terminal, middle, and basal. Flower bud differentiation s tarted in September in the highlands of the Lishan area, and in J uly in the lowlands of the Zhoulan area. In Lishan, flower bud differentiation was higher in the middle and basal segments; during leaf fall, however, flower bud differentiation occurred rapidly in the terminal segment. In Zhoulan, flower buds began to differentiate from the terminal section of the shoot, and severe flower bud necrosis was noted. In July, flower buds developed normally; however, in early August, some of the buds at the basal segment showed browning. During leaf fall, some flower buds showed symptoms of necrosis with rapid and complete browning. Flower bud necrosis began at the basal segment and progressed rapidly towards middle and terminal sections. Before leaf fall, flower buds fell off when scales swelled. The terminal and middle parts of the current-year shoots, with some flower buds, collected in October or later from the Lishan area could be used as scions for top-grafting of 'Shinko' pear. Each grafting scion was a 3-5 cm shoot with one flower bud. These results suggest that scions from the terminal and middle segments of stems of 'Shinko' pear from the Lishan area can be used as scions whereas those from Zhoulan area show necrosis and might not be suitable as scions.

• Journal of Forest and Environmental Science
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• 제25권2호
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• pp.111-118
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• 2009

Multiple shoot induction and plant regeneration using apical bud and nodal explants of 100 year old tree of Anthocephalus cadamba, an important sacred and medicinal tree in India was achieved for the first time. Aseptic explants cultured in Murashige and Skoog (MS) medium augmented with different concentrations of BAP (0.1, 0.5, 1.0, 2.5, 5.0 and 10 mg/l), when maintained for 60 days, healthy shoots were induced in presence of BAP (1 mg/l). Lower concentrations of BAP (0.1 - 0.5 mg/l) induced only one shoot per explant. Increase in number of shoots per explant was observed in presence of higher concentrations of BAP (2.5, 5.0 and 10 mg/l). However, elongation of shoots was completely inhibited. Bud break and shoot regeneration was largely associated with seasonal factors. Apical buds cultured during June to August exhibited early bud break within two weeks of initial culture. In rest of the months, bud break and shoot regeneration was very slow irrespective of the various concentrations of BAP used in the medium. Explants sourced from three different maturity levels of shoots indicated that actively growing shoots from the mother plant with 1 - 2 nodal segments was more suitable for culture initiation than the explants collected from mature shoots at dormant stage. Regenerated shoots with 2 - 3 pairs of leaves when transferred to half strength MS medium fortified with IBA (1 mg/l), 60% of the shoots induced healthy roots, indicating the possibility of large scale micropropagation.

• Plant Biotechnology Reports
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• 제5권3호
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• pp.245-254
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• 2011

This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing $N^6$-Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51.3% bud slices were pflp-transformed, and 12.6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4.4% for pflp and 13.1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems.