• Title/Summary/Keyword: IN VITRO SEED GERMINATION

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In vitro seed germination of Cymbidium aloifolium (L.) Sw., a potential medicinal Orchid from Eastern Ghats of Tamil Nadu, India

  • Philip Robinson, J.;Jyoti, Prasad Kakati;Sebastinraj, J.;Suriya, K
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.343-348
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    • 2017
  • Cymbidium aloifolium (L). Sw. is an exquisite epiphytic orchid of the Kolli Hills (Eastern Ghats) of Tamil Nadu in Southern India. It is fast disappearing from its natural habitats due to deforestation and low germination rate in natural habitat. In the present study, an attempt was made to germinate the seeds from un-dehisced capsule of Cymbidium aloifolium (L). Sw under in vitro condition. The seed germination and protocorm development were recorded in three different well known media namely Knudson C (KC), Half strength Murashige & Skoog (1/2 MS) and Vacin & Went (VW) media. The highest seed germination of 90% was observed KC basal media after $30^{th}$ days whereas germination percentages were 40% and 30% on 1/2 MS and VW media respectively. The well-developed protocorm were transferred to KC media supplemented with 6-Benzyl Amino Purine (BAP) and Naphthalene acetic acid (NAA) where BAP (1.0 mg/l) and NAA (1.0 mg/l) together were found to be optimum for the highest shoot formation. About 90% of the shoots found to be well rooted after transfer to the KC medium differently supplemented with 1.5 mg/l Indole-3-acetic acid (IAA) and 1.0 mg/l Indole-3-butyric acid (IBA). Though rooting also took place in the two basic media but the duration was longer when compared with the hormone-supplemented media. The rooted plantlets were hardened and kept under greenhouse conditions which can be relocated in natural habitats.

In vitro germination of Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama treated by NaOCl (NaOCl 처리에 의한 한라천마(Gastrodia verrucosa Blume)와 애기천마(Hetaeria sikokiana Tuyama)의 기내발아)

  • Bae, Kee-Hwa;Ko, Myoung-Suk;Choi, Sun-A;Lee, Hak-Bong;Kim, Nam-Young;Song, Jae-Mo;Song, Gwan-Pil
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.163-168
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    • 2012
  • An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, $59{\mu}m$, 91% and $138{\mu}m$ through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.

Regeneration of plants from alginate-encapsulated axenic nodal segments of Paederia foetida L. - A medicinally important and vulnerable plant species

  • Behera, Biswaranjan;Behera, Shashikanta;Shasmita, Shasmita;Mohapatra, Debasish;Barik, Durga Prasad;Naik, Soumendra Kumar
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.255-263
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    • 2021
  • Paederia foetida L. is an important medicinal plant that has been used for the treatment of various gastrointestinal related ailments by different tribal communities in India. This plant is also known for its use as a food. Due to overexploitation, P. foetida has been classified as a vulnerable plant in some states of India. The propagation of P. foetida by conventional methods is easy but very slow. Synthetic seed technology offers incredible potential for in vitro propagation of threatened and commercially valuable plants, and can also facilitate the storage and exchange of axenic plant material between laboratories. However, synthetic seed production for P. foetida has not yet been reported. Thus, to the best of our knowledge, the present study is the first attempt to produce synthetic seeds of P. foetida by calcium alginate encapsulation of in vitro regenerated axenic nodal segments. Sodium alginate (3%) and CaCl2 (100 mM) were found to be the optimal materials for the preparation of ideal synthetic seeds, both in terms of morphology and germination ability. The synthetic seeds showed the best germination (formation of both shoot as well as root; 83.3%) on ½ MS medium augmented with 0.5 mg/L indole-3-acetic acid. The plantlets obtained from these synthetic seeds could be successfully acclimatized under field conditions. We also studied the storage of these synthetic seeds at low temperature and their subsequent sprouting/germination. The seeds showed a germination rate of 63.3% even after 21 days of storage at 4 ℃; thus, they could be useful for transfer and exchange of P. foetida germplasm.

Effects of Gamma Irradiation on Nutrient Composition, Anti-nutritional Factors, In vitro Digestibility and Ruminal Degradation of Whole Cotton Seed

  • Hahm, Sahng-Wook;Son, Heyin;Kim, Wook;Oh, Young-Kyoon;Son, Yong-Suk
    • Journal of Animal Science and Technology
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    • v.55 no.2
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    • pp.123-130
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    • 2013
  • Whole cotton seed (WCS) has become one of the major feed ingredients in TMR for dairy cattle in Korea, and WCS for feed use is mostly imported from abroad. Since this genetically modified oil seed is usually fed to the animal in raw state, its germination ability, if last long, often causes concerns about ecological disturbances. In the process of looking for effective conditions to remove germination ability of WCS this study had the objectives to evaluate the nutritional effects of gamma irradiation at doses of 8, 10 and 12 kGy on changes in nutrient contents, anti-nutritional factors, in vitro digestibility and ruminal degradability. No significant differences were found in proximate analysis of nutrients between raw WCS and gamma irradiated one. Glycine and threonine contents significantly increased when the WCS was exposed to gamma ray as compared to untreated WCS (p<0.05). As for fatty acid composition, no significant differences were observed with the irradiation treatment. Free gossypol in WCS was decreased (p<0.05) by gamma irradiation treatment. Of the 3 different levels of gamma irradiation, a dose of 12 kGy was found to be the most effective in reducing free gossypol concentration. Results obtained from in situ experiment indicated that gamma irradiation at a dose of 10 kGy significantly (p<0.05) lowered rumen degradability of both dry matter and crude protein as compared with raw WCS. However, there were no significant differences in rapidly degradable and potentially degradable fractions of crude protein due to 10 kGy gamma irradiation. Overall, this study show that gamma irradiation at a dose of 10 kGy is the optimum condition for removing germination ability of WCS, and could improve nutritive value for the ruminant with respect to the decrease in both ruminal protein degradability and gossypol content of WCS.

Effects of Plant Growth Regulators on In Vitro Germination and Organ Formation of Wild Angelica gigas N. (야생 참당귀(Angelica gigas N.)의 기내발아 및 기관유도에 미치는 PGRs의 영향)

  • Lee, Su-Gwang;Cho, Won-Woo;Lee, Song-Hee;Park, Kwang-Woo;Choi, Kyung;Kang, Ho-Duck
    • Korean Journal of Plant Resources
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    • v.25 no.4
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    • pp.456-464
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    • 2012
  • This study was conducted to establish the in vitro optimal condition for seed germination and organogenesis of wild Angelica gigas. The experiment was evaluated the effects of $GA_3$ for pre-treatment with different periods of time (0h, 24h, 48h, 72h) and followed the treatment of seeds by control, scarification and methanol-heating method. As a result, the highest rates (15%) of seed germination was shown under the treatment without soaking of $GA_3$ and methanol-heating treatment. The seed germination was highly increased 60% under the condition of treatment on ultrasonic waves (frequency 80 KHz) with methanol-heating treatment including 0.1 mg/L $GA_3$. The highest callus induction rate was obtained from in vitro germinated stem, root and hypocotyl on the MS medium with 1.0 mg/L NAA and 0.5 mg/L BA. The highest percentages of shooting (50%) and rooting (85%) induction were observed in hypocotyl and root cultured on PGRs free medium and 0.1 mg/L NAA, respectively. In addition, somatic embryogenesis was observed from stem (1.0 mg/L 2,4-D) and hypocotyl (0.1 mg/L NAA).

Optimization of factors influencing in vitro immature seed germination in Chionanthus retusus

  • Tar, Khin Yae Kyi;Naing, Aung Htay;Ai, Trinh Ngoc;Chung, Mi Young;Kim, Chang Kil
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.347-356
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    • 2018
  • Chionanthus retusus is a small deciduous tree that is widely used in landscaping due to its beautiful white spring flowers and ornamental value. Conventional propagation through seeds requires one to two years of breaking dormancy. The objective of this study was to determine the conditions of in vitro germination in C. retusus. In vitro embryo culture was carried out to investigate the effects of six factors: basal media (McCown Woody Plant Medium (WPM) and Murashige and Skoog (MS)); plant growth regulators (different combinations and concentrations of naphthaleneacetic acid (NAA), 6-Benzylaminopurine (BA), and gibberellic acid ($GA_3$)); embryo age (collected weekly beginning 36 days after fruit setting); low temperature pretreatment (storing $4^{\circ}C$ for 1, 2, 3, and 4 weeks); coconut additives (100, 200, and $300ml{\cdot}L^{-1}$); and genotype (grouping plants depending on their flowering nature). The basal medium used in this study was WPM with $2mg{\cdot}L^{-1-1}\;GA_3$, $20g{\cdot}L^{-1}$ sucrose, and $6g{\cdot}L^{-1}$ Agar. WPM medium mixed with $GA_3$, resulted in higher germination rate as compared to when using a combination of auxin and cytokinin. $GA_3$ at $2mg{\cdot}L^{-1}$ was the most effective of all combinations and concentrations of PGRs. WPM medium with $2mg{\cdot}L^{-1}GA_3$ resulted in better and faster germination (75.93%). Embryos collected at 57 days after fruit setting had the highest percent of germinated seeds (87.04%) while low-temperature pretreatment of fruits at $4^{\circ}C$ for two weeks produced the highest germination (95.37%). These results of this study could be an open ground for development of an efficient protocol for commercial production of the ornamental tree.

Asymbiotic Germination of Korean Native Calanthe Species (한국(韓國) 자생(自生) 새우난초 종자(種子)의 무균발아(無菌發芽))

  • Kim, Chang-Kil;Chung, Jae-Dong
    • Current Research on Agriculture and Life Sciences
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    • v.15
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    • pp.47-52
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    • 1997
  • The experiment was carried out to find the optimal condition for seed germination in vitro of Calanthe species. Seeds of Calanthe spp. after 90 days self pollination were well germinated in the MS medium. Germination condition of cross seeds between C. discolor and C. striata was much improved by pretreatment of micro sonic waves for 10 min. The days of germination was more shortened. and protocorm growth was more promoted in microsonic wave treatment than in non-treatment.

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In vitro seed germination and callus formation on flower bud of Korean mistletoe ( Viscum album L. var. cololatum [Kom.] Ohwi) (겨우살이 종자 발아 및 화아 배양에 의한 캘러스 형성)

  • Kim, Suk-Weon;Ko, Suk-Min;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.47-53
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    • 2008
  • Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.

Effects of altering medium strength and sucrose concentration on in vitro germination and seedling growth of Cypripedium macranthos Sw.

  • Huh, Yoon Sun;Lee, Joung Kwan;Nam, Sang Young;Hong, Eui Yon;Paek, Kee Yoeup;Son, Sung Won
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.132-137
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    • 2016
  • The genus Cypripedium, known as lady's slipper orchid, has been considered to have a considerable marketability as a potted and garden plant with beautiful flowers; however, this species is becoming endangered and, in some places extinct, due to habitat destruction and illegal collections. As such, an optimized artificial propagation system is necessary for its conservation and horticultural cultivation. For the establishment of the in vitro proliferation of the endangered Cypripedium macranthos Sw., native to Korea, the effects of medium strength and sucrose concentrations on germination, protocorm formation and seedling growth were investigated through asymbiotic seed culture. The highest germination rate and protocorm formation rate were obtained with a 1/4 MS medium; higher MS medium strengths did not generate the favorable conditions required for germination and protocorm growth. The overall growth characteristics of roots and buds developed from protocorms were best in cultures of 1/4 MS medium. On this medium containing $10g{\cdot}L^{-1}$ sucrose, both the germination rate and protocorm formation rate increased significantly. The general growth properties of seedlings developed on the medium with $10g{\cdot}L^{-1}$ sucrose were best, showing the highest bud formation rate and root number. Our results demonstrate that the 1/4 MS medium, supplemented with $10g{\cdot}L^{-1}$ sucrose, could improve in vitro germination, and facilitate the growth of seedlings developed from the protocorms of C. macranthos Sw.