Plant Regeneration from Cotyledon Explants in Leaf Chicory

Geun-Won Choi,Dae-Sung Kim,Hyeon-Jeong Hwang,Won Byoung Chae,Youn-Hyung Lee 한국원예학회 2009 Horticulture, Environment, and Biotechnology Vol.50 No.1

Plantlets of leaf chicory ‘Radicchio’ and ‘Italiana’ (Cichorium intybus L. var. foliosum) were produced through organogenesis from cotyledon explants cultured on MS basal medium supplemented with various concentrations of indole-3-acetic acid (IAA) and 6-benzylaminopurine (BAP) combinations. Most vigorous callus and shoot formation of ‘Radicchio’ were observed in the combination of 1.0 ㎎ㆍℓ<SUP>-1</SUP> IAA and 1.0 mgㆍℓ<SUP>-1</SUP> BAP and those of ‘Italiana’ in 0.1 ㎎ㆍℓ<SUP>-1</SUP> IAA and 1.0 ㎎ㆍℓ<SUP>-1</SUP> BAP. Silver nitrate (AgNO₃) which was applied to reduce vitrification was also effective on theformation of single shoots; also, 10 and 5 ㎎ㆍℓ<SUP>-1</SUP> AgNO₃ were selected as the most effective levels in ‘Radicchio’ and ‘Italiana’, respectively. Root formation was promoted when regenerated shoots were cultured in 1/2 MS medium with 0.5 ㎎ㆍℓ<SUP>-1</SUP> naphthaleneacetic acid (NAA) for both ‘Radicchio’ and ‘Italiana’ although adventitious roots were formed without NAA. Rooted plants were transferred into pots and thereafter, acclimatized and maintained in proper greenhouse conditions.

Plant Regeneration from Cotyledon Explants in Leaf Chicory (Cichorium intybus L. var. foliosum)

최근원,김대성,황현정,채원병,이윤형 한국원예학회 2009 Horticulture, Environment, and Biotechnology Vol.50 No.1

Plantlets of leaf chicory ‘Radicchio’ and ‘Italiana’ (Cichorium intybus L. var. foliosum) were produced through organogenesis from cotyledon explants cultured on MS basal medium supplemented with various concentrations of indole-3-acetic acid (IAA) and 6-benzylaminopurine (BAP) combinations. Most vigorous callus and shoot formation of ‘Radicchio’ were observed in the combination of 1.0 mg・L-1 IAA and 1.0 mg・L-1 BAP and those of ‘Italiana’ in 0.1 mg・L-1 IAA and 1.0 mg・L-1 BAP. Silver nitrate (AgNO3) which was applied to reduce vitrification was also effective on the formation of single shoots; also, 10 and 5 mg・L-1 AgNO3 were selected as the most effective levels in ‘Radicchio’ and ‘Italiana’, respectively. Root formation was promoted when regenerated shoots were cultured in 1/2 MS medium with 0.5 mg・L-1 naphthaleneacetic acid (NAA) for both ‘Radicchio’ and ‘Italiana’ although adventitious roots were formed without NAA. Rooted plants were transferred into pots and thereafter, acclimatized and maintained in proper greenhouse conditions.

Callus Induction and Plant Regeneration from Root Explants of Pinellia ternata

Hyun-Soon Kim,Tae-Choon Kim,Seung-Yeob Lee 한국원예학회 2005 Horticulture, Environment, and Biotechnology Vol.46 No.1

To regenerate plants from root segments of Pinellia ternata, an oriental medicinal plant, the effect of growth regulators on callus induction and plant regeneration was investigated from root explants derived from in vitro propagated tuberlets. Adventitious roots from tuberlets showed the highest initiation on Murashige and Skoog (MS) media containing 0.2-0.5 ㎎ㆍL?¹ 1-naphthaleneacetic acid (NAA), but elongation of normal root was achieved on MS basal medium without growth regulators. The highest frequency of callus induction was on MS media supplemented with 1.0 ㎎ㆍL?¹ 2,4-dichlorophenoxyacetic acid and 0.5 ㎎ㆍL?¹ N-6-benzyladenine (BA). For plant regeneration, when calli were transferred to the first media supplemented with 1-4 ㎎ㆍL?¹ BA, most calli became hyperhydric, probably due to higher BA concentrations. At 60 days after transferring to the second regeneration medium containing 1.0 ㎎ㆍL?¹ NAA and 0.25 ㎎ㆍL?¹ kinetin, the frequency of plant regeneration from hyperhydric callus (85-92%) was lower than that from normal callus (100%), according to higher level of BA. Regenerated plants were successfully acclimatized in plastic pots containing a mixture of vermiculite:perlite:leaf-mold (1:1:1), and their survival rate was high.

Adventitious root cultures of Clitoria ternatea L. and its potential as a memory enhancer alternative

Lee Rui Xuan,Hassan Zurina,Subramaniam Sreeramanan,Chew Bee Lynn 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.2

Clitoria ternatea L., known as the butterfy pea plant, is from the Fabaceae family and is commonly used in traditional medicine or as a brain tonic in Asia. The root extracts of C. ternatea have been previously studied for its neuroprotective properties linked to memory enhancement of the mammalian brain, indirectly delaying the occurrence of neurodegenerative diseases. The current study aims to establish in vitro adventitious root cultures of C. ternatea via cotyledon explants and to identify the phytoconstituents of the induced adventitious roots. In this study, the highest percentage of induction (95.24%) was achieved in half-strength MS medium supplemented with 2.50 mg/L NAA (1-naphthaleneacetic acid) after 6 weeks of culture. Results also indicated that the supplementation of 1.50 mg/L NAA induced the highest number of adventitious roots (12.86±2.14), while 2.50 mg/L 4-Cl-IAA (4-chloroindole-3-acetic acid) yielded higher adventitious root length (0.75±0.13 cm) in comparison with the other auxin treatments. Microscopic analysis indicated thicker in vitro root morphology for all NAA treatments in comparison with the roots of in vitro seedling. Ethanolic extract of in vitro-generated adventitious roots showed potential anti-acetylcholinesterase activities (IC50=1519.89±0.18 μg/mL) albeit being lower than the control (IC50=70.37±0.01 μg/mL), while gas chromatography–mass spectrometry analysis identifed the presence of pentacyclic triterpenes, phytosterols and fatty acids in the adventitious root culture that are linked to the mammalian neuroprotective properties of C. ternatea. The current investigation reports on the potential of C. ternatea root cultures being further utilized to produce valuable metabolites linked to the mammalian memory enhancement.

Regeneration of Plantlets from petiole callus of wild viola (Viola patrinii DC.)

Miyake,Hiroshi,Taniguchi,Takeshi,Sato,Tadahiko,Oh,Chang Kwon,Maeda,Eizo 東亞大學校附設遺傳工學硏究所 1996 遺傳工學硏究 Vol.- No.3

Plantlets were regenerated from 5-year subcultured compact callus derived from petiole tissues of wild viola (Viola patrinii DC.) but not from 5-year subcultured friable callus. Regeneration occurred most efficiently on medium that contained two-fold diluted basal salts of Murashige and Skoog's (MS) medium, 5×10-6M 1-naphthaleneacetic acid and ?? M kinetin. The effect of dilution of MS basal salts could also be achieved solely by two-fold dilution of the potassium dihydrogen phosphate in the mixture. The present study revealed that dilution of MS basal salts, in particular of potassium dihydrogen phosphate, was important for the regeneration of wild viola. Moreover, although the callus had been subcultured for 5 years, regeneration of plantlets from callus was still possible. In addition, scanning electron microscopy revealed that details of the process of plant regeneration from subcultured callus varied with the age and source of callus and differed from that reported in rice.

억새(Miscanthus sinensis) 성숙 종자로부터의 캘러스 유도 및 식물체 재분화 체계 확립

조준형,변지희 한국자원식물학회 2011 한국자원식물학회지 Vol.24 No.5

This study was conducted to establish the tissue culture system for Korean domestic Miscanthus sinensis, which is used in various purposes such as forage, and bio-energy resources. With the mature seed of Miscanthus, optimum concentrations of plant growth regulators were identified for an efficient callus induction and regeneration. Among the treatments of 1~10 mg・L^(-1) 2,4-D, IBA, or NAA, callus induction rate was highest (85.3%) on MS medium containing 5 mg・L^(-1) 2,4-D. Under the condition, the callus were efficiently induced and proliferated with comparably lower frequencies of callus browning. In shoot regeneration, the treatment of NAA combined with BAP seemed to contribute more efficient conditions to shoot regeneration than those of NAA with Kinetin or 2-iP. Especially, regeneration efficiency and number of regenerated plants were 83.7% and 5.5 in 3 mg・L^(-1) NAA with 5 mg・L^(-1) BAP, respectively, which were higher frequencies than those in NAA with Kinetin or 2-iP. In results, 5 mg・L^(-1) 2,4-D and 3 mg・L^(-1) NAA combined with 5 mg・L^(-1) BAP were efficient for embryogenic callus induction and regeneration of Miscanthus. This system would be useful for mass-propagation and developing new cultivars via tissue culture of Miscanthus sinensis. 본 연구는 최근 한방자원, 사료자원, 바이오에너지 자원 등 다양하게 이용되는 국내 자생 억새(Miscanthus sinensis)의 대량생산 및 신품종 개발을 위한 조직배양체계 확립을 위해 수행되었다. 이를 위해 억새 완숙종자로부터의 캘러스 유도와 재분화를 위한 식물생장조절제의 적정농도를 규명하였다. 억새의 성숙종자유래 배발생 캘러스 유도를 위해 2,4-D, IBA, NAA를 1~10 mg・L-1의 농도로 단용 처리한 결과, 5 mg・L-1 2,4-D 처리에서 가장 높은 85.3%의 캘러스 유도율과 캘러스의 증식을 보였으며 조직배양 과정 중 갈변화율도 가장 낮았다. 또한, 캘러스의 재분화를 위해 옥신인 NAA와 Kinetin, 2-iP, 또는 BAP 등의 사이토키닌을 혼용 처리한 결과, 각각 19.0%~59.0%, 23.0%~67.3%, 14.7%~83.7%의 재분화율을 보여 NAA와 BAP의 혼용 처리구가 NAA와 Kinetin 또는 2-iP와 혼용 처리구보다 식물체 재분화에 효과적이었다. 특히 3 mg・L-1 NAA와 5 mg・L-1 BAP 혼용 처리된 배지에서의 재분화율이 83.7%로 가장 높게 나타났으며, 캘러스 당 재분화 식물체 개수도 5.5개로 동일농도의 2-iP 또는 Kinetin 혼용 처리 시 2.1 및 2.0개보다 많았다. 본 연구결과 억새 성숙 종자로부터의 배발생 캘러스 유도에는 5 mg・L-1 2,4-D가 그리고 캘러스의 재분화에는 3 mg・L-1 NAA와 5 mg・L-1 BAP 혼용 처리가 가장 효율적이었다. 본 연구를 통해 확립된 조직배양체계는 억새의 대량생산 및 신품종 개발에 유용할 것으로 판단된다.

Influence of Genotype, Explant Source, and Gelling Agent on in Vitro Shoot Regeneration of Chrysanthemum

임기병,권수진,이수인,황윤정,Aung Htay Naing 한국원예학회 2012 Horticulture, Environment, and Biotechnology Vol.53 No.4

The capacity for shoot regeneration of leaf, petiole, and stem explants of eleven chrysanthemum cultivars was examined on the MS medium containing 1 μM naphthaleneacetic acid and 10 μM 6-benzyladenine solidified with 0.8% Agar, 0.4% Agarose, or 0.25% Gelrite. Significant differences in frequency of callus formation and regeneration from the different explants were observed among the different cultivars when grown on the media solidified with the different gelling agents. Gelrite was found to be the most effective gelling agent in promoting of the shoot. Of the different explants used, in general, stem exhibited the highest frequencies of shoot organogenesis and mean number of shoots per explant regardless of cultivar and gelling agent. However, the highest frequency of regeneration (11.67shoots per explant) was noted from leaf explants of cv. Borami followed by (4.33 shoots per explant) from stem explants of cv. Yes Nuri. Shoots were directly developed from the surface of explants, not through callus formation. Low frequencies of shoot organogenesis were observed for the remaining cultivars except for cvs. Yes Time and Yes Star, which exhibits no shoot formation at all. In this study, we have developed an efficient in vitro protocol for cvs. Borami and Yes Nuri from suitable explant.

‘설향’ 딸기 삽목묘의 최적 삽수 채취시기와 오옥신 처리 구명

김은지(Eun Ji Kim),엄미정(Mi Jeong Uhm),정현수(Hyun Soo Jung),김종엽(Jong Yeob Kim),이준구(Jun Gu Lee) (사)한국생물환경조절학회 2020 생물환경조절학회지 Vol.29 No.3

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 육묘시에는 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. The aim of this study was to investigate the effects of timing of collecting date and concentration of IBA and NAA, in order to enhance initial activity and seedling quality of domestic strawberry. Strawberry cuttings were separately taken twice, in June 7 and in July 5, and IBA and NAA were treated with the concentrations of 0.025, 0.05 and 0.1% at cutting date, respectively. The seedlings were evaluated for the percentage of survival during 18 days at 6 times after tunnel cultivation. The NAA treatment was inappropriate for strawberry cutting due to the high rate of seedling mortality, regardless of the collecting date. The vitality of the seedlings was highest at IBA 0.1% in June collecting and at IBA 0.05% in July collecting. The seedlings from June collecting had a higher quantum yield at IBA 0.1% and the seedlings from July collecting at IBA 0.05%. Therefore, IBA could be more effectively applied than NAA to promote the vitality and quality with the appropriate concentration of 0.1% at June collecting and 0.05% at July collecting, respectively.

Rapid Somatic Embryogenesis and Plant Regeneration in American Ginseng: Effete of Auxins and Explants

Wang X.,Proctor J.T.A.,KrishnaRaj S.,Saxena P.K.,Sullivan J.A. The Korean Society of Ginseng 1999 Journal of Ginseng Research Vol.23 No.3

The efficacy of three auxins, viz. 2,4-0, NAA and dicamba, were compared for the induction of somatic embryogenesis in American ginseng (Panax quinquefolium L.). Somatic embryos (SEs) formed on ginseng cotyledonary, zygotic embryo and shoot explants after 8 weeks of induction by the auxin stimuli. Significantly more somatic embryos were induced by culture of any of the ginseng explants on media supplemented with $5{\mu}M$ 2,4-0 than any other auxin treatment. Shoots derived from somatic embryos had the greatest regenerative potential and zygotic embryos the least. Explants generated from green (unstratified) seeds gave similar or higher frequency of embryogenesis as the explants derived from stratified seeds. Histological and SEM studies confirmed that the regenerimts were somatic embryos. Somatic embryos germinated and developed into normal plants in $3\~6$ months. About $10\%$ of plantlets from second generation SEs formed flowers within 10 weeks, particularly on media supplemented with $GA_3$ The development of a regeneration system for ginseng through somatic embryogenesis is a necessary first step for mass propagation and genetic improvement of American ginseng.

적과제 처리가 ‘홍로’와 ‘후지’ 사과의 착과 및 과실품질에 미치는 영향

유진기(Jingi Yoo),박무용(Moo-Yong Park),강인규(In-Kyu Kang) 한국원예학회 2014 원예과학기술지 Vol.32 No.5

본 연구는 ‘홍로’와 ‘후지’ 품종에 benzyladenine(BA, 99% purity), MaxCel<SUP>®</SUP>(1.9% BA), Fruitone(3.5% NAA), MaxCel<SUP>®</SUP> + Fruitone, simazine 등 약제들이 과실의 착과 및 과실품질에 미치는 영향을 조사하였다. BA와 MaxCel<SUP>®</SUP>은 100㎎·L<SUP>-1</SUP> 농도로, Fruitone은 0.1㎎·L<SUP>-1</SUP> 농도로 과실직경이 6㎜인 만개 8일 후에, 그리고 simazine은 400㎎·L<SUP>-1</SUP> 농도로 만개 7일후와 14일 후에 2회 처리하였다. 적과제 처리 후 ‘홍로’ 품종의 정화아의 과총당 착과수는 MaxCel<SUP>®</SUP> + Fruitone, MaxCel<SUP>®</SUP>, simazine 처리구가 각각 1.67, 1.84, 1.81개로 무처리구 2.35개보다 적어 적과효과를 보였고, 특히 무착과 과총 비율이 높아 적과효과가 더 우수하였다. ‘후지’ 품종의 경우 정화아의 과총당 착과수는 MaxCel<SUP>®</SUP> + Fruitone, Fruitone, MaxCel<SUP>®</SUP> 처리구가 1.29, 1.60, 1.76개로 무처리구 2.56개보다 적어 우수한 적과효과를 보였다. 또한, 두 품종 모두에서 MaxCel<SUP>®</SUP> + Fruitone 혼용처리구가 MaxCel<SUP>®</SUP> 단독처리구에 비하여 적과증진효과를 보였다. 그리고 액화아의 경우도 정화아의 결과와 유사한 결과를 보였다. 과중은 ‘홍로’ 품종의 BA 처리구에서, ‘후지’ 품종에서는 BA와 MaxCel<SUP>®</SUP> 처리구에서 증가하였다. 그리고 가용성 고형물 함량은 두 품종 모두에서 BA, MaxCel<SUP>®</SUP>, MaxCel<SUP>®</SUP> + Fruitone 처리 과실들에서 증가하였지만 다른 과실특성들은 차이를 보이지 않았다. Benzyladenine (BA, 99% purity), MaxCel<SUP>®</SUP> (1.9% BA), Fruitone (3.5% NAA), MaxCel<SUP>®</SUP> + Fruitone, a nd s imazine were applied postbloom as fruitlet thinning agents to mature ‘Hongro’ and ‘Fuji’ apple (Malus domestica Borkh.) trees. BA and MaxCel<SUP>®</SUP> were applied at 100 ㎎·L<SUP>-1</SUP> a.i. while Fruitone at 0.1 ㎎·L<SUP>-1</SUP> a.i. and simazine at 400 ㎎·L<SUP>-1</SUP> a.i. All PGRs were applied at 8 days after full bloom (DAFB, 6 ㎜ fruit diameter) in both cultivars, while simazine was treated twice at 7 and 14 DAFB. In ‘Hongro’, the number of total fruit set per flower cluster in terminal buds was 1.67, 1.84, and 1.81 in MaxCel® + F ruitone, MaxCel<SUP>®</SUP>, and simazine applications, respectively, when compared with 2.35 of water control. These reductions in fruit set were mainly attributed to the increased ratio of defruited clusters by the thinning agents. In ‘Fuji’ apple, the number of total fruit set per flower cluster in terminal buds was 1.29, 1.60, and 1.76 in MaxCel<SUP>®</SUP> + F ruitone, Fruitone, a nd MaxCel<SUP>®</SUP>, respectively, when compared with 2.56 of water control in ‘Fuji’ apple. The addition of Fruitone to the MaxCel<SUP>®</SUP> promoted the thinning efficacy in both cultivars, compared to MaxCel<SUP>®</SUP> only. The thinning efficacies were similarly observed with lateral flowers in both cultivars. A significant increase of fruit weight by the postbloom thinning treatments was observed only in the BA application in ‘Hongro’, while the effect was observed in BA and MaxCel<SUP>®</SUP> in ‘Fuji’. While the soluble solids content increased in the BA, MaxCel<SUP>®</SUP> and MaxCel<SUP>®</SUP>+Fruitone treatments in both cultivars, other fruit quality attributes were not affected by the application of post-bloom thinning agents.