Comparative Study of In Vitro Root and Shoot Proliferation from the Node Explants of Asparagus racemosus Willd

2018 ◽  
Vol 35 (1) ◽  
pp. 121-125
Author(s):  
K. K. Dahal ◽  
S. D. Joshi

Asparagus racemosus Willd. locally known as Kurilo or Shatavari is found throughout the tropical and subtropical regions of Nepal. It is a high value herb because of its medicinal and nutritional values which is leading this species decrease in its natural habitat. Hence, to conserve this species, an in vitro tissue culture method of its multiplication has been applied as an effective method of ex situ conservation. In the experiment, although most of the treatments induced highly insignificant number of adventitious shoots, low concentration of IBA (0.1-0.5 mg/l) in combination with relatively higher concentrations of Kinetin (1.0-2.0 mg/l) in MS medium were found to be significantly inducing up to 8.33±1.308 shoots/node. Among the treatments where hormones were used singly, IAA 0.5 and Kinetin 1.0 induced 4.83±1.08 and 4.66±1.43 shots/node respectively. Hence, a protocol for the effective multiplication of this species has been developed which can be used accordingly.

2017 ◽  
Vol 5 (1) ◽  
pp. 27-31
Author(s):  
Pradip Kumar Chaudhary ◽  
Mitesh Shrestha ◽  
Bal Hari Poudel ◽  
Mahesh Kumar Adhikari

Wild edible mushrooms are becoming endangered all over the world. Very few wild edible mushrooms are found in natural habitat. Volvariella bombycina is an edible and medicinal mushroom. The mushroom was collected in natural habitat growing on Populus tree. Mycelium of the mushroom was developed in PDA slant tubes by tissue culture method, incubated at 25°C for 1-2 weeks. Spawn was developed in wheat grains after incubation at 25°C for 2-3 weeks. Substrates were formulated for the development of fruiting bodies by combination of paddy straw, saw dust and rice husk. Fruiting bodies of V. bombycina was cultivated in these substrates after incubation at 28 ± 2°C for 2-4 weeks. The work describes the optimized process for in vitro culture of wild edible mushroom Volvariella bomybycina.Nepal Journal of Biotechnology. Dec. 2017 Vol. 5, No. 1: 27-31


2021 ◽  
Vol 28 (2) ◽  
pp. 172
Author(s):  
Muhammad Dylan Lawrie ◽  
Zulfa Layina ◽  
Della Rosiana Ningtias ◽  
Falah Nur Alifianto ◽  
Ari Indrianto ◽  
...  

Dendrobium capra is an Indonesian endemic orchid species that live in Java. It grows on low altitude with warm climate. D. capra has beautiful small yellow greenish flower that grow in raceme inflorescence. This orchid faces a threat in its natural habitat due to having a long life cycle and a forestry main commodity as a main host thus categorized as Appendix II on CITES list. To address that problem, ex situ conservation approach using in vitro culture method is necessary. Germination enhancement effort using complex organic substances found that 200 ml/l tomato extract gave best germination result. Analysis on D. capra plantlet growth also showed that MS medium produced better plantlet size than NP, VW and KC medium. Supplementing medium with a combination of NAA and TDZ has also successfully induced early flowering within 11 month of culture period. This information is important to achieve successful in vitro culture of D. capra for various purposes.


2018 ◽  
Vol 77 (1) ◽  
pp. 70-79 ◽  
Author(s):  
Erna Karalija ◽  
Sanja Ćavar Zeljković ◽  
Petr Tarkowski ◽  
Edina Muratović ◽  
Adisa Parić

AbstractKnautia sarajevensisis an endemic plant of the Dinaric Alps and is mainly distributed on Bosnian Mountains. Due to the quite large flower heads and easy maintenance, this plant has a potential use as a substitute ornamental plant forK. arvensisin perennial beds. The current study evaluated the germination process in different treatments in an attempt to suppress dormancy and increase germination rate, and to develop a successful protocol for micropropagation. An over 60% germination rate was achieved through cultivation of seeds on MS basal medium with reduced mineral nutrient composition and the absence of sucrose. On the other hand, a below 10% germination rate was achieved with untreated seeds. Suppression of apical dominance was achieved through application of high concentrations of kinetin, apical shoot decapitation or cultivation of shoots in liquid media. Overall, liquid cultures were more successful as a micropropagation system for this plant. Shoots spontaneously developed roots on multiplication treatments and were successfully acclimatized. Moreover, phenolic compound profile was analysed in the light of the possible medicinal potential of this plant. Variable amounts of total phenolic compounds as well as individual phenolics were recorded, according to treatment and solidification of media. An increase in rosmarinic acid content was reported for kinetin treatments and acclimatized plants comparing to mother plants in natural habitat. The present study shows that choice of cytokinin concentration, explant type as well as culture type influences not only shoot proliferation and apical dominance suppression but alsoin vitroproduction of phenolics.


Plants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1657
Author(s):  
Nqobile P. Hlophe ◽  
Adeyemi O. Aremu ◽  
Karel Doležal ◽  
Johannes Van Staden ◽  
Jeffrey F. Finnie

In Africa and Asia, members of the genus Brachystelma are well-known for their diverse uses, especially their medicinal and nutritional values. However, the use of many Brachystelma species as a valuable resource is generally accompanied by the concern of over-exploitation attributed to their slow growth and general small size. The aim of the current study was to establish efficient micropropagation protocols for three Brachystelma species, namely Brachystelma ngomense (endangered), Brachystelma pulchellum (vulnerable) and Brachystelma pygmaeum (least concern), as a means of ensuring their conservation and survival. This was achieved using nodal segments (~10 mm in length) as the source of explants in the presence of different concentrations of three cytokinins (CK) namely N6-benzyladenine (BA), isopentenyladenine (iP) and meta-topolin riboside (mTR), over a period of 6 weeks. The highest (25 µM) concentration of cytokinin treatments typically resulted in significantly higher shoot proliferation. However, each species differed in its response to specific CK: the optimal concentrations were 25 µM mTR, 25 µM iP and 25 µM BA for Brachystelma ngomense, Brachystelma pulchellum and Brachystelma pygmaeum, respectively. During the in vitro propagation, both Brachystelma ngomense and Brachystelma pygmaeum rooted poorly while regenerated Brachystelma pulchellum generally lacked roots regardless of the CK treatments. Following pulsing (dipping) treatment of in vitro-regenerated shoots with indole-3-butyric acid (IBA), acclimatization of all three Brachystelma species remained extremely limited due to poor rooting ex vitro. To the best of our knowledge, the current protocols provide the first successful report for these Brachystelma species. However, further research remains essential to enhance the efficiency of the devised protocol.


Blood ◽  
1967 ◽  
Vol 29 (6) ◽  
pp. 852-858
Author(s):  
YASUSADA MIURA ◽  
FUMIMARO TAKAKU ◽  
KIKU NAKAO

Abstract 1. An in vitro method to observe radiosensitivity of stem cells was developed in the present study. In vivo and in vitro effect of 60Co irradiation on the erythropoietin-induced stem cell differentiation into erythroblasts was observed, using a tissue culture method of polycythemic mouse spleen. Response to erythropoietin was demonstrated by an appearance of heme synthesis and erythroblasts in spleen fragments. 2. A significant correlation between the rate of appearance of erythroblasts and heme synthesis of the spleen fragments was observed. 3. After irradiation, marked impairment of both heme synthesis and production of erythroblasts was observed, yielding D37 values in the vicinity of 70 r in vivo and 120 r in vitro irradiation, respectively. 4. Marked recovery of erythropoietin-induced heme synthesis in the polycythemic mouse spleen was observed 9 days after 300 r irradiation, with an "overshooting" phenomenon on the 12th day.


1935 ◽  
Vol 31 (2) ◽  
pp. 271-272
Author(s):  
G. Barg

The author has shown in a series of experiments that the tissue culture method is quite applicable for the study of anaerobic bacteria and their toxins, especially those of them that are significantly weakened by filtration.


Author(s):  
Justyna Żabicka ◽  
Piotr Żabicki ◽  
Aneta Słomka ◽  
Monika Jędrzejczyk-Korycińska ◽  
Teresa Nowak ◽  
...  

Abstract The paper presents a technique for micropropagation of endangered in Europe and extinct in Poland Pulsatilla vulgaris for ex situ conservation of the genetic resources. Genotype-dependent induction of somatic embryogenesis and rooting was revealed in series of two experiments (I and II) performed under the same experimental conditions. Shoot tips of seedlings were the best explants in both experiments and Murashige and Skoog (MS) medium supplemented with 0.25 or 0.5 mg L−1 BAP was suitable for induction of somatic embryos (SE) and adventitious shoots. Mass SE was obtained in experiment I after explants transfer on ½ MS (2% sucrose) + 0.45 mg L−1 B1 and extending culture to 2–3 months without passages. Rooting of adventitious shoots was a critical point. Out of seven rooting media used in experiment I, only two, ½ MS hormone free (2% sucrose) + 0.45 mg L−1 B1 or MS + 5 mg L−1 NAA + 3.76 mg L−1 B2 resulted in altogether 36.4% rooted shoots. In experiment II, somatic embryogenesis, rooting and acclimatization of adventitious shoots failed. Regenerated plantlets and seedlings converted from SE from experiment I were acclimatized to ex vitro conditions. Both genome size, determined by flow cytometry, and genetic diversity analyzed by ISSR markers, confirmed the compatibility of regenerants from experiment I with P. vulgaris initial seedlings and commercial cultivar. Regenerants obtained in experiment II differed genetically from the regenerants of experiment I and cultivar. Propagated in vitro tissues/organs (SE, adventitious shoots) of P. vulgaris could be a source of material for cryopreservation, artificial seed production and/or for acclimatization of regenerated plantlets and could be used for restoration of the extinct populations. Key Message The micropropagation technique via organogenesis and somatic embryogenesis of endangered in Europe pasqueflower was developed as a tool for species recovery. The critical point is that somatic embryogenesis is genotype-dependent, which affects the repeatability of the experiments and also imposes applying molecular techniques to confirm the genetic fidelity of the regenerants with the initial material and other genotypes.


Author(s):  
Jeanmaire Molina ◽  
William McLaughlin ◽  
Kyle Wallick ◽  
Ronniel Pedales ◽  
Viviane Marcella Marius ◽  
...  

The large-flowered parasitic genus Rafflesia R.Br. (Rafflesiaceae) has long fascinated naturalists and scientists and is an iconic symbol for plant conservation. Techniques to effectively propagate members of the genus outside of their natural habitat are sparse, and grafting infected Tetrastigma K.Schum.(Vitaceae) host plants has previously been reported as a successful strategy for ex situ conservation of Rafflesia. Here we report our attempts in the United States to propagate host cuttings infected with Rafflesia speciosa Barcelona & Fernando and R. lagascae Blanco collectedfrom the Philippines, as well as uninfected host material. We also describe efforts to germinate R. speciosa seeds in vitro using various plant growth regulators (PGRs). After rooting, infected host cuttings survived for a maximum of 11 months, but did not produce shoots. However, an uninfected cutting of T. cf. magnum grafted onto an established Malaysian species of Tetrastigma in June 2017 has succeeded in the commencement of new growth. Three propagules of a second potential host, T. harmandii Planch., have also been vigorously growing at the United States Botanic Garden since June 2017. However, Rafflesia seeds did not germinate with the application of PGRs, even though the seeds were viable according to tetrazolium (TZ) testing.These ex situ propagation attempts have revealed challenges in propagating these species outside of their native ranges, but our incremental success in rooting infected Tetrastigma, as well as grafting interspecific Tetrastigma species, bodes well for further advances. With Philippine host species, T. harmandii and T. cf. magnum in cultivation, we can begin using these specimens for future experimentation involving grafting of infected material and Rafflesia seed inoculation trials.Furthermore, we describe new avenues of propagation techniques for Rafflesia as practised by Marius Gabin, one of the owners of the Vivian Rafflesia garden, which contains a natural Rafflesia forest habitat at Poring Springs, Sabah, Malaysia. Gabin openly shared his successes in artificially inoculating Rafflesia seeds into a mature Tetrastigma vine. Gabin’s willingness to share his experience highlights the importance of collaborating with practitioners who have developed local knowledge of Rafflesia horticulture and conservation.


2016 ◽  
Vol 19 (2) ◽  
pp. 111 ◽  
Author(s):  
Asri Insiana Putri ◽  
Mohammad Na'iem ◽  
Sapto Indrioko ◽  
Sri Rahayu ◽  
Ari Indrianto

In vitro screening of Falcataria moluccana (Miq.) was conducted by tissue culture method. Seeds fromtwo different site of community forest, 400 m (S1) and 800 m (S2) above sea level, were used as material.Double concentration of MS (Murashige & Skoog, 1962) with 40 mg/l gall rust (Uromycladium tepperianum(Sacc.) fi ltrate were used for media selection. The results of this research showed that 66 % axenic plantlets invitro from S1 and 27 % from S2 were still survived after 3 months incubation without subculture. The meanof fresh weight (2. 21 ± 0. 26 g) and dry weight (1. 97 ± 0. 12 g) from S1 plantlets lower than the mean of freshweight (2. 87 ± 0,18 g) and dry weight (2. 16 ± 0. 14 g) from S2 plantlets. Qualitative of terpenes, saponins andquantitative of total phenolics were analyzed from those gall rust extract, as source of fi ltrate media, attackedand un-attacked of F. moluccana. They all qualitatively have capability to produce terpenoid and saponin. Itis notice that U. tepperianum, un-cultured pathogen, contain of those compound that may play a role as codeterminantsof pathogenecity. While the highest total phenolic compound were contained in gall rust extract(2. 35 %), followed by attacked F. moluccana branches (1. 18 %) and un-attacked F. moluccana branches (0. 44%). This indicated that phenolic compound in gall rust has higher activity as a response of F. moluccana to U.tepperianum pathogen pressures and result of this study suggest the great value of gall rust fi ltrate for use asmedia selection in vitro.


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