• Journal of Plant Biotechnology
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• 제39권3호
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• pp.133-139
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• 2012

The influences of ethylene inhibitors ($AgNO_3$ and silver thiosulfate) and cytokinins (BAP and TDZ) on shoot regeneration from cotyledon and hypocotyl explants of B. napus cv. Youngsan were investigated. The presence of $50{\mu}M$ Silver thiosulfate (STS) in shoot regeneration medium formed shoots at 60-68% after 3-4 weeks of culture, which was enhanced by 2-fold compared to that of Silver nitrate ($AgNO_3$). Moreover, cotyledon explants were more regenerative than hypocotyls; shoots from cotyledon explants began to occur 4-5 days earlier than that of hypocotyl explants. TDZ at a concentration of $8-10{\mu}M$ was effective for shoot regeneration, compared with BAP. Consequently, the optimal shoot regeneration response was observed in medium supplemented with $50{\mu}M\;STS+8{\mu}M\;TDZ$. In transmission electron microscopy (TEM) analysis, higher density of silver nanoparticles was shown to be accumulated widely inside the cell wall and plasmodesmata of regenerating leaf cultured in medium supplemented with $AgNO_3$. By contrast, in the cell cultured in medium with STS, fine-grained deposits were partly observed in the surroundings of the cell wall.

• 식물조직배양학회지
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• 제21권5호
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• pp.315-318
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• 1994

CaMV35S promoter-$\beta$-glucuronidase (GUS) 유전자와 선발 표지로서 neomycin phosphotransferase 유전자를 가진 pBI121 binary 벡터를 도입한 Agrobacterium turmfaciens LAB4404와 더덕 유식물체의 자엽 절편을 1 mg/L BA가 첨가된 MS 배지에서 48시간 동안 공동배양한 후 1 mg/L BA, 250 mg/L carbenicillin 100 mg/L kanamycin sulfate을 첨가한 고체배지에 옮겨 명조건에서 배양하였다. 배양 2주 후 절편의 절단면 부근으로부터 수많은 부정아가 형성되기 시작하였다. 이들 부정아의 GUS 활성을 조사한 결과 15%가 양성반응을 나타내었다. 배양 6주 후 절편이 형성한 수많은 부정아로부터 56.7% 빈도로 shoot이 발달하였다. 이들 shoot은 기본배지에 옮겨서 4주 경과되었을 때 대부분 발근하였으며, 재분화된 개체는 토양에 이식하였다. Southem 분석결과 GUS양성을 보인 재분화 개체의 게놈 DNA에 GUS 유전자가 삽입되었음이 확인되었다.

• Journal of Plant Biotechnology
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• 제48권1호
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• pp.54-61
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• 2021

This work describe an efficient method for the shoot induction and plant regeneration of seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim. The highest rate of shoot induction (82.2%) was obtained when apical bud explants from juvenile seedlings (5 months old) were cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BAP). However, apical bud explants obtained from mature trees (12 years old) did not produce any shoots, even with BAP supplementation. Among the three cytokinins tested for shoot multiplication (BAP, zeatin, and kinetin), BAP was the most effective; the highest number of shoots per explant (2.1) was observed on MS medium supplemented with 1.0 mg/L BAP. In contrast, the longest average shoot length (3.0 cm) was observed after growth on MS medium with 2.0 mg/L zeatin. No multiplication occurred when apical bud explants were cultured with kinetin-supplemented media. During rooting of in vitro-elongated shoots, the highest rooting rate (100%) was observed in half-strength MS medium supplemented with 0.5 ~ 1.0 mg/L indole-3-butyric acid (IBA) or 3.0 mg/L 1-naphthaleneacetic acid (NAA). During the acclimatization process, plantlets that were rooted on the IBA (0.5 mg/L)-supplemented medium had the highest survival rate (100%) and maximum root length (18.5 cm). These findings suggest that a low concentration (0.5 mg/L) of IBA is appropriate for the rooting and acclimatization of T. mandshurica. Plants were successfully transferred to the greenhouse with a 100% survival rate. This protocol will be useful for the large-scale propagation of Tilia species.

• Journal of Plant Biotechnology
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• 제49권1호
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• pp.61-73
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• 2022

In this study, we developed a reliable and efficient Agrobacterium-mediated genetic transformation system by applying sonication and vacuum infiltration to six chickpea cultivars (ICCV2, ICCV10, ICCV92944, ICCV37, JAKI9218, and JG11) using embryo axis explants. Wounded explants were precultured for 3 days in shoot induction medium (SIM) before sonication and vacuum infiltration with an Agrobacterium suspension and co-cultivated for 3 days in co-cultivation medium containing 100 µM/l of acetosyringone and 200 mg/l of L-cysteine. Responsive explants with putatively transformed shoots were selected using a gradual increase in kanamycin from 25 mg/l to 100 mg/l in selection medium to eliminate escapes. Results showed optimal transformation efficiency at a bacterial density of 1.0, an optical density at 600 nm wavelength (OD₆₀₀), and an infection duration of 30 min. The presence and stable integration of the β-glucuronidase (gusA) gene into the chickpea genome were confirmed using GUS histochemical assay and polymerase chain reaction. A high transformation efficiency was achieved among the different factors tested using embryo axis explants of cv. JAKI 9218. Of the six chickpea cultivars tested, JAKI9218 showed the highest transformation efficiency of 8.6%, followed by JG11 (7.2%), ICCV92944 (6.8%), ICCV37 (5.4%), ICCV2 (4.8%), and ICCV10 (4.6%). These findings showed that the Agrobacterium-mediated genetic transformation system will help transfer novel candidate genes into chickpea.

• Journal of Plant Biotechnology
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• 제2권3호
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• pp.151-155
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• 2000

Shoot proliferation was obtained from shoot tips and nodal explants of Vitex negundo L. on MS medium supplemented with either BAP or KIN (0.1-2.0 mg/L) alone or in combination with NAA (0.1 mg/L). The concentrations of cytokinins combined with NAA produced multiple shoots from shoot tips and nodal explants. The highest mean percentage (84.3$\pm$8.0) of shoot multiplication's were observed on nodal explants in the presence of BAP (1.5 mg/L) and NAA (0.1 mg/L) followed by shoot tips (65.0$\pm$5.0). The regenerated shootlets were rooted on MS basal medium IAA, IBA, NAA (0.1-1.5 mg/L). The maximum number of roots (51.0$\pm$2.6) was achieved on the medium containing IBA (1.0 mg/L) followed by other auxins (NAA, IAA). The regenerated plants were successfully transferred to a mixture of vermiculate and soil. About 95% of the plantlets survived when transferred to the field.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.51-55
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• 2006

A rapid micropropagation protocol was established for Exacum travancoricum Bedd. The effect of two cytokinins viz. BA and kinetin were studied to evaluate the propagation of plants through nodal explants. MS medium supplemented with 13.32 ${\mu}M$ BA induced early bud break and subsequent production of multiple shoots. Rooting of shoots occurred when cultured on 1/2 strength MS medium supplemented with 14.7 ${\mu}M$ IBA. Rooted plants were acclimatized to greenhouse conditions. The propagated plants were transferred successfully to field with 65% success. As the plant was amenable to propagation in vitro, this can be employed as a tool for conservation of this critically endangered and endemic ornamental herb.

• Journal of Life Science
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• 제9권1호
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• pp.50-53
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• 1999

Effects of growth regulators and medium components were tested for shoot multiplication and callus growth from shoot explants of black locust. During shoot multiplication, callus growth at the cut end of shoot explants proceeded shoot development. The basal callus growth seemed to be a function of both mineral components and cytokinin supplemented in the medium. Maximum callus growth was induced by 0.5${\mu}$M BAP and the callus growth decreased as the level of BAP increased. Positive correlations were found between basal callus growth, and shoot multiplication and growth. Shoot multiplication was greatest on BSM medium (black locust shoot culture medium) supplemented with 1 $\mu$M BAP. With medium containing high nitrogen content, both shoot multiplication and growth were significantly enhanced. A new BRM medium was the most effective for rooting of black locust among three rooting media tested.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.151-154
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• 2003

This study was conducted to determine the optimal concentrations of plant hormones (2,4-D, picloram, dicamba, NAA, kinetin) and the suitable explants among seeds, hypocotyl, and cotyledons on calls formation and plant regeneration of stevia(Stevia rebaudiana Bertoni). The frequency of cellus formation was higher in the young leaf-explants then the older ones, and in the seeds then the hypocotyls and cotyledons on MS medium with 1mg/L 2,4-D. After transfer of seed-derived stevia callus producing embryogenic callus on plant-regeneration medium, the frequency of plant regeneration from callus was 23.8% in MS medium with 1mg/L NAA and 3mg/L kinetin.

• Plant Resources
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• 제6권2호
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• pp.89-93
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• 2003

This study was conducted to develop the clonal propagation technique through in vitro culture using basal stem explants in Phalaenopsis hybrid grown in vitro. The highest frequency of protocorm-like body (PLB) formation was obtained when basal stem explants were cultured on VW medium containing 30g/L sucrose, 500 mg/L activated charcoal, 150 ml/L coconut water, 1 mg/L NAA, 5 mg/L 2iP and 2.5g/L gel rite. PLBs transferred to Hyponex medium were regenerated to plantlets. Plantlets transferred to plastic pots containing spagnum moss were developed and successfully acclimatized under greenhouse. The flower was bloomingly opened in plants regenerated from basal stem explants. The flower was not different from both mother plant and plant induced through clonal propagation of Phalaenopsis hybrid.

• Journal of Plant Biology
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• 제38권4호
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• pp.365-371
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• 1995

Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.