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.

scholarly journals An Improved Micropropagation Protocol byEx VitroRooting ofPassiflora edulisSims. f.flavicarpaDeg. through Nodal Segment Culture

Scientifica ◽  
2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari ◽  
C. P. Ravindran
Keyword(s):  
Survival Rate ◽  
Clonal Propagation ◽  
Basal Medium ◽  
Multiple Shoots ◽  
Nodal Segment ◽  
Ms Medium ◽  
Ex Vitro ◽  
Passiflora Edulis ◽  
Half Strength

A procedure for rapid clonal propagation ofPassiflora edulisSims. f.flavicarpaDeg. (Passifloraceae) has been developed in this study. Nodal explants were sterilized with 0.1% HgCl2and inoculated on Murashige and Skoog (MS) basal medium. The addition of 2.0 mgL−16-benzylaminopurine (BAP) to MS medium caused an extensive proliferation of multiple shoots (8.21±1.13) primordial from the nodal meristems. Subculturing of these multiple shoots on the MS medium augmented with 1.0 mgL−1of each BAP and Kinetin (Kin) was successful for the multiplication of the shootsin vitrowith maximum numbers of shoots (25.73±0.06) within four weeks of incubation. Shoots were rooted best (7.13±0.56roots/shoots) on half strength MS medium supplemented with 2.0 mgL−1indole-3 butyric acid (IBA). Allin vitroregenerated shoots were rooted byex vitromethod, and this has achieved 6-7 roots per shoot by pulsing of cut ends of the shoots using 200 as well as 300 mgL−1IBA. The plantlets were hardened in the greenhouse for 4-5 weeks. The hardened plantlets were shifted to manure containing nursery polybags after five weeks and then transferred to a sand bed for another four weeks for acclimatization before field planting with 88% survival rate.

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 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

88 EFFECT OF VITRIFICATION PROCEDURE ON SURVIVAL RATE OF BOVINE EMBRYOS PRODUCED IN VITRO

2011 ◽  
Vol 23 (1) ◽  
pp. 149
Author(s):  
E. Y. Herrera ◽  
C. de Frutos ◽  
R. Laguna-Barraza ◽  
A. Gutierrez-Adan ◽  
D. Rizos
Keyword(s):  
Survival Rate ◽  
Survival Rates ◽  
Calf Serum ◽  
Basal Medium ◽  
Bovine Embryos ◽  
Oviduct Fluid ◽  
Bovine Oocytes ◽  
Cryopreservation Method ◽  
Hatching Rates

Vitrification as a cryopreservation method has many advantages compared with slow freezing. Many variables in the vitrification process exists that influence the survival rates of vitrified oocytes and embryos. These include the cryoprotectants (type, concentration, and duration of exposure), the temperature of the vitrification solution at exposure, the device used for vitrification, and the quality and developmental stage of embryos. It is worthwhile to mention that vitrification protocols successfully used in bovine oocytes and embryos have been used also with human oocytes and embryos. Vitrification is relatively simple, requires no freezing equipment, and relies on the placement of the embryos in a very small volume of vitrification medium that must be cooled at extreme rates not obtainable in regular enclosed straws. The aim of the present study was to evaluate the efficiency of 4 different vitrification protocols on the survival rate of in vitro produced (IVP) bovine embryos. Blastocysts were produced by a standard IVP procedure following in vitro maturation, fertilization, and culture in synthetic oviduct fluid supplemented with 5% fetal calf serum (FCS). On Day 7 (Day of IVF = Day 0), a total of 297 blastocysts were vitrified using (i) the open pulled straw (OPS) in 20% DMSO and 20% ethylene glycol (EG) in a basal medium of TCM-199 with HEPES supplemented with 20% FCS; (ii) the modified OPS, in 20% DMSO, 20% EG, and 0.5 M sucrose in a basal medium of phosphate buffer saline (PBS) supplemented with 20% FCS; (iii) the cryoloop, in 15% DMSO, 15% EG, 10 mg mL–1 Ficoll 70, and 0.65 M sucrose in a basal medium of PBS supplemented with 20% FCS; and (iv) in 0.25 straws in 20% glycerol, 20% EG, 0.3 M sucrose, 3% polyethylene glycol, and 0.3 M xylose in a basal medium of PBS. After warming, embryos were placed in culture for additional 24 h. Re-expansion and hatching rates were measured at 2 and 24 h after warming. Data were analysed by 1-way ANOVA. At 2 h post-warming, the re-expansion of blastocysts vitrified with cryoloop was significantly higher compared with OPS, modified OPS, and the 0.25 straw methods (54.08 ± 15.53 v. 10.40 ± 3.00, 22.67 ± 9.20, and 8.82 ± 2.15, respectively; P ≤ 0.028). At 24 h post-warming, only embryos from cryoloop and modified OPS were still alive with a survival rate of embryos vitrified with cryoloop significantly higher than that of those vitrified with modified OPS (48.45 ± 17.56 v. 3.75 ± 3.75, respectively; P ≤ 0.007). Hatching rates at 24 h post-warming were not different between cryoloop and modified OPS groups (5.63 ± 4.40 and 1.25 ± 1.25, respectively). These results clearly demonstrate that embryo cryotolerance is affected by the method used for cryopreservation. Moreover, cryoloop vitrification was found to be more effective than OPS and 0.25 straw methods for the cryopreservation of bovine embryos.

scholarly journals Micropropagation of Clerodendrum serratum L. through Direct and Indirect Organogenesis

2013 ◽  
Vol 22 (2) ◽  
pp. 179-185 ◽  
Author(s):  
SM Vidya ◽  
V Krishna ◽  
BK Manjunatha ◽  
MR Pradeepa
Keyword(s):  
Survival Rate ◽  
Medicinal Plant ◽  
Clonal Propagation ◽  
Growth Hormones ◽  
Direct Organogenesis ◽  
Maximum Frequency ◽  
Indirect Organogenesis ◽  
Half Strength ◽  
In Vitro Clonal Propagation

In vitro clonal propagation of Clerodendrum serratum L., a rare medicinal plant has been reported by using LM medium supplemented with different growth hormones. The maximum number of shoots with maximum length were obtained from stem derived callus on LM media fortified with 1.5 mg/l BAP and 0.3 mg/l NAA. Nodal explants showed direct organogenesis on LM media containing BAP (0.5 mg/l) alone. The regenerated shoots were successfully rooted with maximum frequency (100%) on half strength LM media supplemented with 0.5 mg/l NAA. The well rooted microshoots were successfully transferred to hardening and survival rate was 88%. DOI: http://dx.doi.org/10.3329/ptcb.v22i2.14208 Plant Tissue Cult. & Biotech. 22(2): 179-185, 2012 (December)

Hyperhydricity reversal and clonal propagation of four-wing saltbush (Atriplex canescens, Chenopodiaceae) cultivated in vitro

2008 ◽  
Vol 56 (4) ◽  
pp. 358 ◽  
Author(s):  
Isaac Reyes-Vera ◽  
Carol Potenza ◽  
Jerry Barrow
Keyword(s):  
Culture Medium ◽  
Clonal Propagation ◽  
Inorganic Nitrogen ◽  
Basal Medium ◽  
Control Treatment ◽  
Culture Vessel ◽  
Cumulative Frequency ◽  
Atriplex Canescens ◽  
Normal Morphology

In vitro propagated shoots of four-wing saltbush (Atriplex canescens, Pursh Nutt) showed severe symptoms of hyperhydricity. We show that the reversion of hyperhydric A. canescens shoots to normal shoots was significantly affected by the presence of inorganic nitrogen in the culture vessel. When the culture vessel was vented or when ammonium nitrate was deleted from Murashige and Skoog basal medium, rates of reversion were significantly higher. Although statistically significant differences were evident when comparing vented v. non-vented treatments for each medium, the modified culture medium with vented closures was consistently the best treatment, showing a total cumulative frequency of 39.7% reversion to normal morphology, compared with a total cumulative frequency of 7.1% observed in the control treatment. Resulting normal shoots also showed significant improvements in further manipulations, including rooting in vitro, transplantation to soil and survival in native sites.

scholarly journals Micropropagation of Turbinicarpus valdezianus (Möeller) Glass & Foster (Cactaceae) an Endemic Cactus in Northern Mexico

HortScience ◽  
2016 ◽  
Vol 51 (1) ◽  
pp. 94-97 ◽  
Author(s):  
Alejandro Martínez Palacios ◽  
Raúl Cárdenas Navarro ◽  
Diana Beatriz Hernández Ortega ◽  
Víctor Chávez Avila
Keyword(s):  
Clonal Propagation ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Inhibitory Effect ◽  
Multiplication Rate ◽  
Adventitious Shoot Formation ◽  
In Vitro Clonal Propagation

An in vitro clonal propagation protocol based on axillary bud development was generated for Turbinicarpus valdezianus. An efficient multiplication rate was obtained using either longitudinal or apical explants from in vitro germinated seedlings. The proliferation capacity of these explants was evaluated by testing the single and interaction effects of five concentrations of 6-furfurylaminopurine (KIN) (0.00, 2.32, 4.64, 9.28, and 18.56 µm) and three concentrations of α-naphthalenacetic acid (NAA) (0.00, 0.54, and 2.70 µm), using Murashige and Skoog (MS) as basal medium. Statistical analysis showed that the highest average shoot proliferation of T. valdezianus was recorded with 9.28 µm of KIN, producing 11.75 and 4.50 plantlets per initial explant, for apical and lateral explants, respectively. Addition of NAA to the medium had an inhibitory effect on shoot proliferation for both explant types. The developed shoots in 9.28 µm of KIN and plant growth regulator (PGR)-free treatments were used for a rooting subculture phase. These shoots were then transferred to PGR-free MS medium, resulting in statistically significant different rooting frequencies of 78% and 97%, respectively. When transplanted in soil, the rooted shoots showed an average survival rate of 90%, without any significant statistical differences between treatments. This propagation protocol has the capacity to produce near to 21 plantlets per seedling in 27 weeks, i.e., 11.78 and 9.00 plantlets per apical and lateral explants, respectively, without callus or adventitious shoot formation. These features made it highly attractive as an in vitro clonal propagation method for T. valdezianus plants and the later implementation of a rescue program for threatened wild populations of this cacti species.

scholarly journals In Vitro Clonal Propagation of Scoparia dulcis L., a Perennial Medicinal Herb

1970 ◽  
Vol 44 (3) ◽  
pp. 341-346
Author(s):  
AKM Sayeed Hassan ◽  
Laila Shamroze Bari ◽  
Rebeka Sultana ◽  
Nadira Begum ◽  
Rahima Khatun
Keyword(s):  
Clonal Propagation ◽  
Basal Medium ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Medicinal Herb ◽  
Naphthaleneacetic Acid ◽  
Scoparia Dulcis ◽  
Half Strength ◽  
In Vitro Clonal Propagation

An efficient protocol was established for in vitro clonal propagation of the perennial medicinal herb Scoparia dulcis L. (Family. Scrophulariaceae) through in vitro culture. Apical and axillary buds of young sprouts from selected plants were used as explants. Best shoot induction was observed on MS basal medium supplemented with 0.1 mg/l BAP, in which 94% of the explants produced 12 shoots per culture. Repeated subcultures in the same medium, resulted rapid shoot multiplication with 16 shoots per culture. The half strength MS medium with 0.5 mg/l IBA +0.5 mg/l NAA the highest percentage (85.20) and maximum number (13.40) of roots were initiated within four weeks of culture. For acclimatization and transplantation, the plantlets in the rooting culture tubes were kept in normal room temperature for 7 days before transplanting in pots where plantlets were reared for three weeks. The survival rate of regenerated plantlets was 85%. Key words: Scoparia dulcis, Medicinal plant, Shoot proliferation, Micropropagation, Acclimatization, IAA (indoleacetic acid), IBA(indolebutanoic acid), NAA(α-naphthaleneacetic acid), BAP(benzylamino purine) DOI: 10.3329/bjsir.v44i3.4408 Bangladesh J. Sci. Ind. Res. 44(3), 341-346, 2009

scholarly journals Micropropagation of Paphiopedilum x Dalatense

2021 ◽  
Vol 19 (1) ◽  
pp. 155-163
Author(s):  
Tran Thai Vinh ◽  
H’ Yon Niê Bing ◽  
Dang Thi Tham ◽  
Nguyen Thi Thanh Hang ◽  
Vu Kim Cong ◽  
...  
Keyword(s):  
Survival Rate ◽  
Humic Acid ◽  
Shoot Formation ◽  
In Vitro Rooting ◽  
Ms Medium ◽  
Orchid Species ◽  
Best Response ◽  
Yeast Powder ◽  
Half Strength

Paphiopedilum x dalatense is a beautiful orchid species with large flowers in variable colors and leaves covered with stripes and beautiful unseen mosaic spots. Recently, many people exploit this species, causing it becomes very rare. In this study, we studied the effects of various organic matter: potato, banana and tryptone, yeast powder, peptone on the growth and development of P. dalatense shoots as well as the effects of NAA and humic acid on in vitro rooting of this orchid were investigated. The research results showed that MS medium supplemented with 100 g/L banana in combination with 100 g/L potato (5,4 shoots/sample, 18,8 mm/shoot, 4,5 leaves/shoot, and shoots survival rate of 100%) or MS medium supplemented with 1 g/L peptone (4,19 shoots/sample, 15 mm/shoot, 4 leaves/bud, and 92% of shoots survival rate) were the best response for the shoot formation and development. In addition, the half strength MS culture medium supplemented with 1 mg/L NAA (5,2 leaves/sample, 4,6 roots/buds, 3,56 cm/root, and 100% rate for rooting) was the suitable medium for the in vitro rooting of P. dalatense. Being cultured on half strength MS medium supplemented with 2 mg/L humic acid, the rooting rate reached 100% with the greatest root number and the longest root (5 roots/shoots, 5,5 cm/root). The obtained results on the in vitro propagation on this orchid helps contribute to the conservation and increases the genotic pool of this precious wild orchid species, as well as the rapid multiplication of healthy plantlets serving the commercialization of precious orchid species.

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