scholarly journals Organic Additives Improves the in Vitro Growth of Native Orchid Vanda helvola Blume

2015 ◽  
Vol 7 (2) ◽  
pp. 192-197
Author(s):  
Devina DAVID ◽  
Roslina JAWAN ◽  
Hartinie MARBAWI ◽  
Jualang Azlan GANSAU
Keyword(s):  
Seed Germination ◽  
Rapid Development ◽  
Basal Medium ◽  
Tomato Juice ◽  
Organic Additives ◽  
Orchid Species ◽  
Orchid Conservation ◽  
Hand Pollination ◽  
Basal Media

In vitro seed germination has been proven to be the most efficient technique to propagate orchid. The application of this aseptic technique has contributed to conservation of many endangered orchid species. In this study, undehisced capsules of Vanda helvola Blume were collected from Orchid Conservation Centre in Lagud Sebrang Agriculture Park, after 120 days from hand pollination and aseptically cultured on three types of basal media such as Murashige and Skoog (MS), Knudson C (KC) and Vacin and Went (VW). After 90 days of culture, 66.40 ± 4.14% of seeds successfully germinated on KC medium. The effect of organic additives such as tomato juice, coconut water, peptone and yeast extract at different level of concentrations in KC basal medium were also tested on seed germination and seedling development of this native orchid. After 90 days of culture, over 90% of seeds were tremendously germinated on KC medium supplemented with 10% or 15% (v/v) of tomato juice. The incorporation of peptone at 0.1% (w/v) in KC basal media promoted rapid development of protocorm to seedling. Seedlings on this treatment produced an average of three leaves and two roots after 90 days of culture and were successfully acclimatized.

scholarly journals A Review on Asymbiotic Seed Germination in Orchids through Plant Tissue Culture

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Purnima Paramanik ◽  
◽  
Dipak Kar ◽  
Subrata Raha ◽  
◽  
...  
Keyword(s):  
Seed Germination ◽  
Mineral Nutrients ◽  
Symbiotic Association ◽  
Organic Additives ◽  
Asymbiotic Seed Germination ◽  
Orchid Species ◽  
Mycorrhizal Association ◽  
Commercially Important

Orchid seeds are very small, dust like in appearance, fusiform in shape, lacking endosperm and have undifferentiated embryo covered by transparent seed coat. Mycorrhizal association is required for seed germination of naturally growing orchids. In this symbiotic association, orchid species are dependent on mycorrhizal association for supply of mineral nutrients. In in-vitro condition, such demand of minerals may be compensated by external supply of sugar and mineral nutrients that are required for seed germination of orchid. Several orchids are responded by this asymbiotic seed culture and have commercial importance. Therefore, formulation of efficient in-vitro protocol is important for commercially important orchid species as well as endangered orchid species for conservation. This review paper is focused on various aspects of asymbiotic seed germination of orchids and the role of organic additives in successful seed germination.

scholarly journals Effects of Media composition on Asymbiotic Seed Culture of Vanda tessellata : An Approach to In vitro Conservation

2020 ◽  
Vol 30 (2) ◽  
pp. 285-295
Author(s):  
Sumana Aditya ◽  
Tustu Mondal ◽  
Nirmalya Banerjee
Keyword(s):  
Survival Rate ◽  
Seed Germination ◽  
Significant Variation ◽  
Basal Medium ◽  
Common Phenomenon ◽  
Media Composition ◽  
Basal Media ◽  
Effects Of Media ◽  
Vitamin Mixture

For initiation of seed germination and protocorm growth in Vanda tessellata (Roxb.) Hook. ex G. Don different media, namely Knudson’s C basal medium, modified Knudson’s C basal medium, Murashige and Skoog basal medium, Vacin and Went basal medium and Lindeman basal medium either alone or supplemented with vitamin or 0.1% peptone (w/v) were used. Vitamin mixture was consisted of nicotinic acid, pyridoxine and thiamine HCl at 1 : 1 : 10, respectively. For the initiation of germination, all the treatments exhibited promising results without showing significant variation. But the rate of survival of the germinated seedlings was remarkably low in all the basal media. Maximum survival rate of germinated seedlings was recorded in MS + 0.1% peptone. Necrosis of protocorms was a common phenomenon in all the treatments and 100% necrosis was recorded in LM basal medium and LM + vitamin. Addition of vitamin mixture and peptone in the basal media increased the rate of survival as well as differentiation of the germinated protocorms. Maximum rooted plantlets were recorded in VW + 0.1% peptone Plant Tissue Cult. & Biotech. 30(2): 285-295, 2020 (December)

scholarly journals In vitro seed germination in Cymbidium elegans Lindl. and Dendrobium densiflorum Lindl. ex Wall. (Orchidaceae)

2010 ◽  
Vol 6 ◽  
pp. 100-102 ◽  
Author(s):  
Shreeti Pradha ◽  
Bijaya Pant
Keyword(s):  
Seed Germination ◽  
Basal Medium ◽  
Naphthalene Acetic Acid ◽  
Genetic Constitution ◽  
Ms Medium ◽  
Orchid Species ◽  
C Elegans ◽  
Protocorm Formation ◽  
Dendrobium Densiflorum

A comparative study of in vitro seed germination of two endangered orchid species, viz. Cymbidium elegans Lindl. and Dendrobium densiflorum Lindl. ex Wall., was carried out on Murashige and Skoog's (MS) medium, supplemented with different concentrations and combination of 6-benzylaminopurine (BAP) and á-Naphthalene acetic acid (NAA). The hormone-free MS medium and MS medium supplemented with various growth hormones were found effective for in vitro seed germination of both species. However, the seeds of these two species showed variation in their germination behavior. Hormone-free MS basal medium was found most effective for seed germination of D. densiflorum; whereas, basal medium supplemented with BAP (1mg/l) was effective for C. elegans. The seeds of D. densiflorum showed quick response in earlier germination, protocorm formation and further development into seedlings in comparison to C. elegans. In C. elegans, germination of immature seeds started after nine weeks of inoculation; whereas in D. densiflorum, the initiation of germination started after five weeks of culture. The variations in seed germination, protocorm formation and seedling differentiation in the two orchid species might be due to the differences in their genetic constitution and the presence of different endogenous growth stimulating substances present in their seeds. The present study has provided useful information for in vitro clonal mass multiplication of these commercially important orchid species. Key-words: growth hormone; in vitro study; orchid.DOI: 10.3126/botor.v6i0.2917 Botanica Orientalis - Journal of Plant Science (2009) 6: 100-102

scholarly journals Germination and Protocorm Formation of Ophrys Sphegodes Mill. – In Vitro Protocol for a Rare Orchid Species

2018 ◽  
Vol 67 (3-4) ◽  
pp. 196-201
Author(s):  
Jovana Dulić ◽  
Mirjana Ljubojević ◽  
Ines Prlainović ◽  
Goran Barać ◽  
Tijana Narandžić ◽  
...  
Keyword(s):  
Seed Germination ◽  
Germination Rate ◽  
Casein Hydrolysate ◽  
Habitat Destruction ◽  
Ex Situ ◽  
Organic Additives ◽  
Orchid Species ◽  
Short Period ◽  
Protocorm Formation

Summary Ophrys sphegodes Mill. is a wild orchid species which is threatened and protected due to its pollination biology, small seed and habitat destruction. The aim of this study was to establish asymbiotic germination protocol for the purpose of ex situ conservation. Two basal media Knudson C (KC) and Malmgren (MM), supplemented with organic additives (peptone (PE), L-glutamin (A)e, folic acid, casein hydrolysate (CA)) added separately and control media KC--C and MM--C were used in the present research. All the nutrition media contained 2% sucrose, 7% agar and 1% activated carbon, while their pH was adjusted to 5.8 ± 0.02 before autoclaving at 121 ° C for 20 minutes. The seeds were examined under two illumination conditions, 0/24 light/dark (L/D) and 16/8 L/D. The presented results indicate a huge influence of illumination and nutrition media on the seed germination and protocorm formation. The seed germination was overall significantly more successful in dark conditions (0/24 L/D) than with lighting (16/8 L/D). Protocorm, rhizoids and shoot formation were achieved only on the seeds cultured on MM medium, while the KC medium caused only swelling of the embryo. Organic additives had positive influence on the germination rate. According to the obtained results, the best germination rate and seedling development were achieved on MM-PE media, cultured in dark. The presented procedure accelerates the germination period and can provide a large number of plants in a relatively short period of time so it can be used for conservation programs and mass production protocol.

In vitro propagation of the bay laurel (Laurus nobilis.L) in Morocco

2014 ◽  
Vol 4 (3) ◽  
pp. 96-103
Author(s):  
Abdelali Chourfi ◽  
Tajelmolk Alaoui ◽  
Ghizlane Echchgadda
Keyword(s):  
Seed Germination ◽  
In Vitro Culture ◽  
In Vitro Propagation ◽  
Aerial Part ◽  
Basal Medium ◽  
Axillary Buds ◽  
Laurus Nobilis

Laurus nobilis L. is among the species which are most threatened by massive degradation in Morocco. The multiplication by seed or by cuttings gives very low percentages of recovery that is insufficient to meet the demand of growing market. In vitro culture proves to be a tremendous asset to solve this problem. Our work has focused on the study of seed germination of this species and its multiplication from microcuttings. Finally, we studied the ac-climatization ability of the plantlets resulting from this germination. The study of the germination, via the further measurement of the length of the aerial part and the roots and the number of axillary buds for nine weeks, showed that the MS basal medium was more efficient than media 1/2M.S and WPM. Among the eight tested hormones, IAA yielded the best growth of the plantlets. Hormonal combination of NAA and kinetin resulted into a per-centage of the greatest success in reaching 67 % micropropagation. The study also revealed that the MS basal medium in the presence of the IAA plants can acclimate most easily in two types of substrates with improved development in the peat alone.

scholarly journals Establishment of an EfficientIn VitroRegeneration Protocol for Rapid and Mass Propagation ofDendrobium chrysotoxumLindl. Using Seed Culture

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Potshangbam Nongdam ◽  
Leimapokpam Tikendra
Keyword(s):  
Acetic Acid ◽  
Seed Germination ◽  
Germination Percentage ◽  
Naphthalene Acetic Acid ◽  
Mass Propagation ◽  
Asymbiotic Seed Germination ◽  
Orchid Species ◽  
Seed Culture ◽  
Leaf Formation

An efficientin vitroregeneration protocol from seed culture has been established successfully forDendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated morein vitrorooting, though IBA was found to be more effective in rooting induction as compared to NAA. Thein vitroprotocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly importantDendrobiumorchid species.

A classroom exercise in hand pollination and in vitro asymbiotic orchid seed germination

2008 ◽  
Vol 93 (2) ◽  
pp. 223-230 ◽  
Author(s):  
Philip J. Kauth ◽  
Timothy R. Johnson ◽  
Scott L. Stewart ◽  
Michael E. Kane
Keyword(s):  
Seed Germination ◽  
Hand Pollination ◽  
Classroom Exercise ◽  
Orchid Seed ◽  
Orchid Seed Germination

scholarly journals ASYMBIOTIC GERMINATION OF CYPRIPEDIUM CALCEOLUS VAR. PARVIFLORUM (ORCHIDACEAE) IN VITRO.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 618d-618
Author(s):  
Chin-Chang Chu ◽  
Kenneth W. Mudge
Keyword(s):  
Seed Germination ◽  
Suspension Culture ◽  
Agar Medium ◽  
Germination Rate ◽  
Coconut Water ◽  
Mature Seed ◽  
Orchid Species ◽  
Cypripedium Calceolus ◽  
Chilling Treatment

Seeds from mature seed pods of Cypripedium calceolus var. parviflorum were germinated on 1/4 MSMO (Sigma) + 100ml/l coconut water + 1% sucrose +/- 8g/l agar (pH 6.0), and with or without prechilling at 5C for 8 weeks. Protocorm with apex (stage 3) was use as an index of germination. Seeds sown on agar medium withou chilling treatment resulted in a 40% germination rate in 120 days but the germination was very uneven. Seeds germinated on agar medium with prechilling developed more synchronously with 92% germination in 60 days (ie. about 120 days after sowing). Suspension culture of seeds without prechilling resulted in 85% germination after 90 days. The synchronization of seed germination in suspension culture was intermediate between that on agar with and without prechilling. Protocorms germinated in suspension culture appeared morphologically identical to those germinated on agar medium. All stage 3 protocorms developed further on the same agar medium in darkness. Both agar and suspension culture in media containing coconut water provided reliable seed germination methods for this orchid species.

scholarly journals Micropropagation of Cypripedium formosanum Hayata through Axillary Buds from Mature Plants

HortScience ◽  
2010 ◽  
Vol 45 (9) ◽  
pp. 1369-1372 ◽  
Author(s):  
Yung-I Lee
Keyword(s):  
Average Length ◽  
Basal Medium ◽  
Shoot Multiplication ◽  
Shoot Formation ◽  
Axillary Buds ◽  
Orchid Species ◽  
Optimum Result ◽  
Aseptic Culture ◽  
Slipper Orchid

A micropropagation protocol for an endangered slipper orchid species, Cypripedium formosanum Hayata, through axillary buds from adult plants has been developed. The season of explant collection is crucial for the initial success of an aseptic culture. Explants collected in the middle of January gave the highest percentage of explant survival (54.2%) and shoot-forming percentage (41.7%). Of the two cytokinins tested, N6-benzyladenine (BA) was found to be superior to thidiazuron for normal shoot formation. The optimum result was obtained in quarter-strength Murashige and Skoog medium containing 22.2 or 44.4 μM BA in which the cultures produced 6.3 and 7.1 shoots per explant with 10.6 to 11.7 mm average length after 90 d of culture. Regenerated shoots rooted for 60 d in the basal medium with 1 g·L−1 activated charcoal and 20 g·L−1 potato homogenate were ready for growth in pots. This is the first report on shoot multiplication in vitro from mature plants of Cypripedium that provides a reliable method for propagating the selected elites.

scholarly journals Clonal propagation of Trichocentrum stramineum (Orchidaceae), a threatened species endemic to Mexico

2020 ◽  
Vol 98 (2) ◽  
pp. 355-365
Author(s):  
Selene Ramos-Ortiz ◽  
Luz Ma. Rangel-Guerrero ◽  
Martha Elena Pedraza-Santos ◽  
Víctor Manuel Chávez-Ávila ◽  
José Guadalupe Martínez-Ávalos ◽  
...  
Keyword(s):  
Survival Rate ◽  
Clonal Propagation ◽  
Basal Medium ◽  
Botanical Garden ◽  
Genetic Characteristics ◽  
Orchid Species ◽  
Apical Bud ◽  
Whole Plants ◽  
Study Species

Background: Cloning techniques are applied to an endangered orchid species in order to reproduce individual plants and to preserve their genetic characteristics. Objectives: To establish a new protocol for clonal propagation of a threatened orchid of horticultural importance. Study species: Trichocentrum stramineum, a threatened orchid endemic to Mexico. Study site and duration: Totutla, Veracruz, Mexico. All experiments were designed and carried out at the Botanical Garden-UNAM and the IIAF-UMSNH over a course of six years. Methods: Seeds were germinated in a modified KC basal medium; protocorms and apical bud explants were obtained from the resulting in vitro plants and cultivated with or without plant growth regulators (PGRs). Both experimental groups were subcultured in order to evaluate the number of protocorm-like bodies (PLBs) and buds per explant. Results: On average, protocorms generated 51.2 and 54.1 PLBs in the absence or presence of 1 mg l-1 6-benzyladenine (BA), respectively, while 13.1 and up to 23.7 PLBs and / or shoots were observed on the apical bud explants in the absence or presence of 1 mg l-1 kinetin, respectively. In both cases, responses were direct, without the formation of an intervening callus. Approximately 200 PLBs were subcultured and developed into whole plants within 14 weeks. These were acclimatized to greenhouse conditions with a 90 % survival rate after 12 weeks. After 44 weeks, flowering was observed (3 %) individuals measuring at least 12 cm in height. Conclusions: The developed protocol proved to hold great potential for commercial propagation and conservation programs.

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