scholarly journals Micropropagation of Phalaenopsis amabilis var. ‘Manila’ by Leaves Obtained from in Vitro Culturing the Nodes of Flower Stalks

2016 ◽  
Vol 8 (2) ◽  
pp. 164-169 ◽  
Author(s):  
Khosro BALILASHAKI ◽  
Masood GHASEMI GHEHSAREH

Orchids are one of the most popular plants in the world and among them the species of Phalaenopsis have the most sales on the global market. Because of its difficult propagation, micropropagation has been suggested recently. In the current study, the leaves obtained from in vitro culture of flower stalk nodes were used as explants and were cultured on MS medium with different concentrations of NAA, BA and TDZ. Protocorm-like bodies (PLBs) were produced and transferred to medium without growth regulators. Finally, the adaptation of plants was evaluated in a medium of cocopeat + coal (3:1 v/v) and another medium of cocopeat + charcoal + LECA (2:1:2 v/v). Results showed that the highest percentage of active samples was 100% which could regenerate the PLBs by being treated with 4 mg/l TDZ. The lowest active samples (60%) were those treated in the medium with 4 mg/l BAP + 0.5 mg/l NAA. The highest PLBs per explant (50.65) were obtained in the medium supplemented with 15 mg/l BAP + 3 mg/l NAA. Best acclimation (90%) of plants was obtained when medium of cocopeat + charcoal + LECA (2:1:2 v/v) was used. According to the results of the current experiment, the MS culture medium containing 15 mg/l BAP + 3 mg/l NAA was thereby considered as the best medium for Phalaenopsis micropropagation.

2020 ◽  
pp. 1887-1893
Author(s):  
Rasha K. Mohammed Al-Saedi ◽  
Ansam G. Abdulhalem

     The current study aimed to adopt a method for inducing callus cells and regenerating the important common red bean using different types of growth regulators such as N6-benzylaminopurine (BAP), Naphthalene acetic acid (NAA), and Thidiazuron (TDZ). Different types of common bean pinto cultivar explants, such as  internodes, cotyledons and roots,  were inoculated on Murashige and Skoog medium (MS) provided with different combinations of plant growth regulators, including 1- BAP (5 mg/l) 2-BAP (4.5 mg/l) NAA (0.5 mg/l), 3- BAP (4.5 mg/l), and TDZ (0.1mg/l). Callus was initiated on MS culture medium supplied with 5 mg/l BAP for all explants (internodes, cotyledons, and roots) at 50, 20, and 10% respectively, while adding NAA with 0.5mg/l showed a low percentage of callus (30%) only in the internode explants. Optimum results were obtained by growing the internodes on MS medium with 4.5 mg/l BA and either 0.5 mg/l NAA or 0.1 mg/l TDZ, transplanting the derived shoots into internodes and cotyledons with 70 and 10% respectively. This study concludes that the internodes as explants have the best growth results.


2017 ◽  
pp. 50-55
Author(s):  
Khalid Lagram ◽  
Mohamed Ben El Caid ◽  
Souad El Aaouam ◽  
Mohamed Lachheb ◽  
Abdelhamid El Mousadik ◽  
...  

Crocus sativus L. is a male sterile vegetatively propagated plant. Its flower produces stigmas that when dried, constitute the source of a spice commonly known as Saffron. Slow vegetative propagation and diseases limit the production and the development of saffron. “In vitro” culture could be an effective method to overcome these limitations by improving the quantity and the quality of the planting materials. In this work, Crocus sativus L. segments corms of cultivar from the region of Taliouine (Southeast of Morocco) were used for the propagation through indirect organogenesis. To optimize the in vitro growth conditions, we have used the Murashige and Skoog medium (MS medium), supplemented with 2.4-dichlorophenoxyacetic acid (2.4-D) and with 6-benzylaminopurin (BAP) at combination of various concentrations. Our results showed the formation of callus in 85.42% of explants that grow in a culture medium supplemented with 2,4-D combined with BAP, at a concentration of 1mg/l each. In addition, we observed that increasing the concentration of BAP in the culture medium to 1.5mg/l improved the rate of shoots initiation (0.81). In the meantime, we noted that a combination of BAP (8mg/l) and Naphthalene acetic acid (NAA; 2mg/l) has significantly improved the rate of the formation of advanced shoots (6.65). Finally, the shoots that developed were transferred to an induction medium of roots and corms. As a result, we observed that 50% of shoots tested in ½ MS medium supplemented with 2.4-D and of BAP (1 mg/l each) and 5% sucrose, formed corms. Our study provides a first database for in vitro culture of Moroccan saffron cultivars.


2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES


2020 ◽  
Vol 19 (4) ◽  
pp. 41-51
Author(s):  
Tour Jan ◽  
Ikram Ullah ◽  
Bilal Muhammad ◽  
_ Tariq ◽  
Ali Mansoor ◽  
...  

Hyperhydricity is a frequently problem in plants during in vitro culture and affected micropropagation ofplants. To develop an efficient in vitro regenerated system without hyperdydricity, we demonstrated the effectof different disinfected agents (mercuric chlorite and hypochlorite), growth regulators, their concentrationsand combinations, Agar, pH, ammonium nitrate (NH4NO3) and number of subcultures. Mercuric chlorite at0.07% and exposing time (9–10 min) was appropriate for hygienic culture. The shoots induced by Benzyladnine(BA) alone or in combination with α-Naphthaleneacetic acid (NAA) exhibited maximum multiplicationwith symptoms of hyperhydricity than those induced by Kinetin alone or in combination with NAA. Hyperhydricitywas also reduced by increasing the concentration of agar, pH and elimination of NH4NO3 from themacroelements of Murashig and Skoog (MS) medium. Repeated subcultures affected both multiplication andhyperhydricity. The multiplication of shoots increased from parental culture up to 5th subculture and thereafterdeclined in 6th subculture. Although shoot hyperhydricity were observed from 1st subculture (19%) andthen increased up to 85% in 6th subculture. This increased in hyperhydricity could be due to the remaininginfluence of hormones. In shoots of 5th subculture the content of chlorophyll (dark green) were higher thanshoots of 6th subculture.


Author(s):  
Diem Thi Le ◽  
Mai Thi Bach Vo

Dendrobium officinale Kimura et Migo has been known for a long time as a precious orchid, which is used for medicine and functional foods that are widely commercialized in the world. The nodal explants could be obtained in high efficiency, good quality and uniform seedlings on multiplication vegetative in vitro. They can be propagated in large quantities in a short time. The studied results showed that the nodal explants grew on MS medium + 10 % coconut milk + 25 g sucrose + 0.5 mg/L BA + 6 g agar/ liter create high buds. This buds were used to A B 1 cm 1,5 cm create bud clusters with the aim of improving in vitro vegetative. The best bud culture medium for the formation of clusters was MS + BA 2.0 mg/L + NAA 0.4 mg/L; MS + kinetin 1.5 mg/L + NAA 0.3 mg/L; MS + TDZ 2.0 mg/L + NAA 0.4 mg/L; MS + adenine 1.5 mg/L + NAA 0.3 mg/L. However, the addition of kinetin or TDZ to the culture medium needed a steps to extend the bud subculture after 45 days in culture. The best medium for the rooting is MS + NAA 0.5 mg/L.


2019 ◽  
Vol 9 (4) ◽  
pp. 676-678
Author(s):  
I. D. Borodulina ◽  
A. Trufanova ◽  
G. Shevchenko ◽  
G. Sokolova ◽  
T. Plaksina

Micropropagation of Chrysanthemum is an alternative to the traditional method of reproduction. Thanks to this method, the Chrysanthemum reproduction time is reduced to 3-4 months. For clonal micropropagation, sterile microshoots of Chrysanthemums of the varieties Snow White, Stranger, and Baltica White were used. At the stage of the micropropagation, the Murashige-Skoog (MS) medium with the full and half composition of mineral salts and growth regulators (kinetin, 6-benzylaminopurine, β-indolylacetic acid) were used. A universal culture medium for clonal micropropagation of all varieties of Chrysanthemum and optimal mediums, taking into account variety-specificity were established. It was noted that under in vitro conditions, high proliferative activity was observed in Neznakomka variety.


Author(s):  
Thi Tinh Nguyen ◽  
Tien Dung Nguyen ◽  
Xuan Thanh Dao ◽  
Truc Dat Chu ◽  
Xuan Binh Ngo

P. vietnamenseO. Gruss & Perner is one of the endemic lady’s slipper orchids in Vietnam that has been known as an endangered species threatened with extinction due to over-collection. To protectP. vietnamensespecies,In Vitropropagation method needs to be established for producing plantlets that can be introduced back in wild, as well as commercialized. In this study, we examined the effect of basal culture medium, plant growth regulators, and AC on the micropropagation ability ofP. vietnamense, which were collected from wild population in Thai Nguyen province, Northeastern Vietnam. We determined that the 0.2% HgCl2can be used in within 5 to 10 min for sterilizing axillary buds ofP. vietnamense, resulting in 65.55% of disinfected samples. Moreover, the 1/2 MS medium supplemented with 2.0 mg 1-1BA and 1.0 mg 1-1NAA, which gave about 2.85 shoots/sample, was the most suitable for shoot regeneration and multiplication. In addition, the result also indicated that 0.5 mg 1-1NAA and 1.0 g 1-1AC gave the best results (88.89%) for root induction and plantlets


Hoehnea ◽  
2018 ◽  
Vol 45 (4) ◽  
pp. 663-668
Author(s):  
Daniel da Silva ◽  
Angela Maria Imakawa ◽  
Suely de Souza Costa ◽  
Paulo de Tarso Barbosa Sampaio

ABSTRACT The aim of this study was to evaluate the in vitro germination of zygotic embryos and seeds of Caesalpinia ferrea Martius and the morphogenetic responses of the explants to different concentrations of growth regulators. Seeds and zygotic embryos were inoculated in MS culture medium and kept in a growth room at a temperature of 25 ± 2 ºC for 16 hours of photoperiod for 30 days. The seeds had a higher in vitro germination rate than the explants from zygotic embryos. However, zygotic embryos in MS medium supplemented with 0.9 mg L-1 BAP had the highest percentage of regeneration (50%), number of shoots (3.25), buds (2.85) and leaves (3.15), multiplication rate (27.75), and length of shoots (1.96 cm). The in vitro culture of zygotic embryos and seeds made possible the multiplication of a higher number of healthy seedlings. Thus, it can be used as an alternative technique for the propagation of this species.


Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 320
Author(s):  
Nisar Ahmad Zahid ◽  
Hawa Z.E. Jaafar ◽  
Mansor Hakiman

Ginger (Zingiber officinale Roscoe) var. Bentong is a monocotyledon plant that belongs to the Zingiberaceae family. Bentong ginger is the most popular cultivar of ginger in Malaysia, which is conventionally propagated by its rhizome. As its rhizomes are the economic part of the plant, the allocation of a large amount of rhizomes as planting materials increases agricultural input cost. Simultaneously, the rhizomes’ availability as planting materials is restricted due to the high demand for fresh rhizomes in the market. Moreover, ginger propagation using its rhizome is accompanied by several types of soil-borne diseases. Plant tissue culture techniques have been applied to produce disease-free planting materials of ginger to overcome these problems. Hence, the in vitro-induced microrhizomes are considered as alternative disease-free planting materials for ginger cultivation. On the other hand, Bentong ginger has not been studied for its microrhizome induction. Therefore, this study was conducted to optimize sucrose and plant growth regulators (PGRs) for its microrhizome induction. Microrhizomes were successfully induced in Murashige and Skoog (MS) medium supplemented with a high sucrose concentration (>45 g L−1). In addition, zeatin at 5–10 µM was found more effective for microrhizome induction than 6-benzylaminopurine (BAP) at a similar concentration. The addition of 7.5 µM 1-naphthaleneacetic acid (NAA) further enhanced microrhizome formation and reduced sucrose’s required dose that needs to be supplied for efficient microrhizome formation. MS medium supplemented with 60 g L−1 sucrose, 10 µM zeatin and 7.5 µM NAA was the optimum combination for the microrhizome induction of Bentong ginger. The in vitro-induced microrhizomes sprouted indoors in moist sand and all the sprouted microrhizomes were successfully established in field conditions. In conclusion, in vitro microrhizomes can be used as disease-free planting materials for the commercial cultivation of Bentong ginger.


2021 ◽  
Vol 2 (2) ◽  
pp. 538-553
Author(s):  
Natacha Coelho ◽  
Alexandra Filipe ◽  
Bruno Medronho ◽  
Solange Magalhães ◽  
Carla Vitorino ◽  
...  

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.


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