• Journal of Plant Biotechnology
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• 제7권4호
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• pp.247-257
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• 2005

An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.

• Plant Biotechnology Reports
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• 제4권4호
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• pp.261-267
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• 2010

We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.

• Journal of Plant Biotechnology
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• 제6권3호
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• pp.171-179
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• 2004

Hormonal requirements inducing different regeneration pathways with particular emphasis on somatic embryo-genesis in Alhagi graecorum were studied. While combination of 0.5 $\mu{M}$ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.5 $\mu{M}$ 6-benzylaminopurine (BAP) and 5 $\mu{M}$ 1-naphthaleneacetic acid (NAA) in MS medium induced callus formation and callus maintenance from internodal explants, each alone or in combination with other induced distinct regeneration pathway. Adventitious bud formation was induced on MS medium supplemented with 2.5 $\mu{M}$ BAP. It was improved when 2.5 $\mu{M}$ BAP was used in combination with 5 $\mu{M}$ NAA. MS medium containing 0.5 $\mu{M}$ 2,4-D or 5 $\mu{M}$ NAA induced the formation of abnormal direct somatic embryos. While increase of 2,4-D concentration (1.125-9) resulted in the formation of viable embryogenic mass, increase of NAA did not change its effect. NAA should be used in combination with 2,4-D even at low concentration (0.5 $\mu{M}$) to form embryogenic mass. In A. gaecorum, the role of 2,4-D as trigger of somatic embryogenesis and BAP as trigger of adventitious bud formation was deduced, but for maximum yield certain auxin-cytokinin ratio should be applied. Embryogenic masses characterized by high water content, low peroxidase activity, and low number of peroxidase and glutamate oxaloacetate transaminase bands in comparison with calli obtained under conditions stimulating adventitious bud formation. The resulted differential gene expression, which could be detected by native-PAGE patterns, could be used as marker for organogenic pathway in A. graecorum.

• 원예과학기술지
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• 제33권2호
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• pp.251-258
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• 2015

Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with $3.0mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 $NH{_4}^+:NO{_3}^-$ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of $50g{\cdot}L^{-1}$ and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, $14.70g{\cdot}L^{-1}$ dry biomass, $10.33mg{\cdot}g^{-1}$ DW of phenolics and $3.89mg{\cdot}g^{-1}$ DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.

• 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.

• Plant Biotechnology Reports
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• 제3권3호
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• pp.243-250
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• 2009

In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production of ${\beta}-carboline$ alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium supplemented with $5.0{\mu}M$ 6 benzyl adenine (BA) and $2.5{\mu}M$ ${\alpha}-naphthaleneacetic$ acid (NAA). The embryogenic callus was maintained on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was at a maximum ($18.1{\pm}0.9$ per g of callus) on MS medium containing $5.0{\mu}M$ BA and $2.5{\mu}M$ NAA together with $75mg\;1^{-1}$ casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation of ${\beta}-carboline$ alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus accumulated equivalent amounts of harmaline ($66.4{\pm}0.5{\mu}g/g$ dry weight), harmine ($82.7{\pm}0.6{\mu}g/g$ dry weight), and diosgenin ($170.7{\pm}1.0{\mu}g/g$ dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals.

• Journal of Plant Biotechnology
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• 제43권4호
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• pp.486-491
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• 2016

본 연구의 목적은 배 '원황'의 효율적인 재분화 조건을 규명하기 위해 NAA 0.01, 0.05, 0.1, 0.5 mg/L과 BA 3, 5, 10 mg/L을 조합하여 처리한 결과, NAA 0.05 mg/L, BA 3 mg/L의 조성에 plant agar 8 g/L와 sucrose 30 g/L의 MS 배지에서 재분화율이 13.3%로 나왔다. 기존의 '신고'와 '황금배'의 발근에 사용하였던 배지 조성과 '배연3호'의 발근 배지조성에서는 '원황' 품종의 발근이 전혀 이뤄지지 않았다. 식물생장조절물질이 '원황'의 발근에 미치는 영향을 알아보기 위하여 NAA 0.5, 1.0 mg/L, IBA 0.3, 0.5, 1.0 mg/L을 조합하여 처리한 결과 IBA 1.0 mg/L와 NAA 1.0 mg/L의 조합에 1/4 LS vitamin이 포함된 배지에서 발근률이 24%로 나타났으며 효율적인 '원황'의 발근을 위해서는 탄소원으로 glucose 7.5 g/L가 사용되었다.

• 생물환경조절학회지
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• 제29권3호
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• pp.252-258
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• 2020

본 연구는 국내 '설향' 딸기 품종의 삽목 육묘 시 초기 활착을 증진시키고 묘소질을 향상시키기 위한 삽수 채취시기와 오옥신계 식물생장조절제 IBA, NAA의 적정 처리농도 구명을 위하여 수행되었다. 딸기 삽수는 6월 7일과 7월 5일의 두 차례에 걸쳐 채취했으며, IBA와 NAA는 각각 0.025, 0.05, 0.1%의 농도로 침지 처리하였다. 삽목한 묘는 터널재배 후 18일 동안 6회에 걸쳐 생존율을 조사하였다. 삽수 채취시기와 관계없이 NAA 처리에서는 묘 고사율이 높아, 딸기 삽목 육묘 시에는 NAA 처리가 부적합하였다. 삽수 채취시기와 식물생장조절제가 묘 생육에 미치는 영향을 조사한 결과 6월 채취묘는 IBA 0.1%에서, 7월 채취묘는 IBA 0.05%에서 묘의 활력이 우수하였다. 광합성 효율을 조사한 결과 6월 채취묘는 IBA 0.1%에서, 7월 채취묘는 IBA 0.05%에서 양자수율이 높았다. 따라서, 딸기 삽목묘의 생육을 증진시키기 위해서는 NAA보다 IBA를 사용하는 것이 적절하며, 이때 6월 삽수 채취 시에는 0.1%, 7월 삽수 채취 시에는 0.05%의 농도로 이용하는 것이 보다 적합한 것으로 판단되었다.

• 한국자원식물학회지
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• 제21권3호
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• pp.184-191
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• 2008

This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators, and total nitrogen and sucrose contents on the in vitro regeneration of Abeliophyllum distichum Nakai. The effects of growth regulators on regeneration from 3 explant sources (leaf, internode and node) were more or less same. Leaf explants produced only callus with 2ip (Isopentenyladenine) and NAA (Naphthaleneacetic acid) treatment and other regulators had no effects. Test with internode explants yielded about same results but callus was obtained with 2,4-D (2,4-Dichlorophenoxyacetic acid). Node explants resulted in shoot regeneration by all regulator treatment except NAA and 2,4-D, but control also showed similar results. Callus formation from internode and node explants was vigorous by 2ip, zeatin, and 2,4-D treatments and high NAA concentration resulted in higher callus formation. In this experiment, various mixed treatment of growth regulators were also employed, using node as explant material. Shoot regeneration was obtained with BA (Benzyl adenine) + NAA treatments but the results were comparable with control. Generally shoot and root regeneration was poor with all combined treatment except 2ip + NAA and 2,4-D + NAA. However, callus was formed readily with all treatments. In this experiment, combined treatments of regulators were applied on the callus derived from singular regulator treatment. The results showed no shoot and root regeneration with any combination of 2,4-D, IAA (Indoleacetic acid) and NAA, but soft milky white callus was formed in all the treatments. No shoot and root regeneration was observed with any combination of 2iP, NAA and IAA, but somewhat hard, light green callus was formed in all the treatments. Callus formation decreased with high kinetin concentration in case of kinetin + NAA treatment. The experiments with total nitrogen content of media showed that low concentrations of 15 and 30mM were effective for the shoot and root regeneration. Sucrose experiment demonstrated shoot regeneration with 1${\sim}$4% concentration, and root and callus formation with 2${\sim}$4%. No root and callus formation was observed with 0 and 1% sucrose.

• 식물조직배양학회지
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• 제24권6호
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• pp.329-334
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• 1997

미류나무의 잎절편들을 fructose를 첨가한 WPM에서 배양하였으나, TDZ와 BA 그리고 NAA의 여러 농도와의 조합에서 다른 형태의 기관들(뿌리나 줄기)이 관찰되었으며, 체세포 배와 유사한 기관들의 형태도 관찰되었다. 3계통 미류나무들의 잎절편들을 사이토카이닌(BA 또는 TDZ)와 오옥신(NAA)를 첨가한 WPM 배지에서 접종시켜, 재분화를 유도한 결과, BA (05 mg/L)나 TDZ (0.01 mg/L)와 NAA (0.05 mg/L 또는 0.1 mg/L)의 조합에서 재분화 율은 각각 63%에서 100%이고, 재분화 개체 수는 잎절편당 평균 2.5부터 20개, 그리고 부정줄기 형성체군(organogenic site) 들은 평균 1.8에서 4.5개로 높게 나타났다. 또한 세 계통의 미류나무들은 각기 유사한 처리 조합에서 높은 재분화율을 나타내었다.