In vitro propagation of the threatened plant Sphaerophysa kotschyana (Fabaceae): inter simple-sequence-repeat (ISSR) analysis and salt tolerance of the regenerants

2013 ◽  
Vol 61 (1) ◽  
pp. 67 ◽  
Author(s):  
Semiha Erişen ◽  
Zeynep Öncel

Sphaerophysa kotschyana is a threatened endemic species in Turkey and according to the Bern Convention, it is on the absolute preservation plant list. In vitro propagation methodologies were evaluated as an ex situ conservation strategy for this species. Nodal segments were cultured on Murashige and Skoog (MS) media with different cytokinins (benzyladenine, thidiazuron (TDZ) and zeatine), with or without auxin (α-naphthaleneacetic acid; NAA), to investigate shoot initiation. TDZ produced the highest number of shoots (11.0 shoots per explant) on MS medium at a concentration of 0.05 mg L–1. Rooting reached 100% when 0.5 mg L–1 NAA was combined with half strength MS and 1.5% sucrose and rooted plantlets were successfully acclimatised. Somaclonal variation of a mother plant and 10 regenerants was assessed using ISSR analysis. The same banding profiles were exhibited by all plants. In vitro response to salinity stress (NaCl) was also investigated in this halophytic species. Higher concentrations of NaCl negatively affected shoot multiplication, whereas shoot height was enhanced at 50 mM NaCl. These results suggest that the established protocol is an efficient and reliable system of in vitro propagation for ex situ conservation of S. kotschyana.

2011 ◽  
Vol 107 (3) ◽  
pp. 531-540 ◽  
Author(s):  
Katerina Grigoriadou ◽  
Nikos Krigas ◽  
Eleni Maloupa

Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.


2015 ◽  
Vol 2 (2) ◽  
pp. 65
Author(s):  
Yusuf Sigit Ahmad Fauzan ◽  
Edhi Sandra ◽  
Daru Mulyono

The population density of natural agarwood (Aquilaria beccariana) in Indonesia decreased to less than one tree per hectare. Efforts have been carried out on ex situ conservation of agarwood despite facing many obstacles. In vitro propagation is one alternative to speed up the recovery of natural agarwood populations. The purpose of this study was to obtain optimal elongation media for in vitro culture with addition of auxin and cytokinin, namely IBA, BAP and kinetin. The results showed that the best auxin-cytokinin combination was IBA 0.1 mg/L and BAP 0.05 mg/L. This combination increased the height and number of segments of A. beccariana with an average height of 1.64 cm and average number of sections of 6.40. It is suggested that this combination of IBA and BAP was the most effective compared to the other treatments. In addition, the combination of IBA 0 mg/L and BAP 0.03 mg/L gave rise to the best response to increase the number of shoots with an average of 1.91 shoots.Keywords: Aquilaria beccariana, shoot, elongation, auxin, cytokinin ABSTRAKKepadatan populasi gaharu (Aquilaria beccariana) alam di Indonesia kurang dari satu pohon per hektar. Upaya pelestarian gaharu ex situ telah banyak dilakukan tetapi masih banyak kendala. Perbanyakan gaharu in vitro merupakan salah satu cara alternatif untuk mempercepat pemulihan populasi gaharu alam. Tujuan penelitian ini adalah untuk memperoleh media elongasi yang optimal pada kultur in vitro gaharu dengan penambahan kombinasi zat pengatur tumbuh auksin dan sitokinin. Pada penelitian ini digunakan auksin IBA, serta sitokinin BAP dan Kinetin. Hasil penelitian elongasi diperoleh kombinasi auksin dan sitokinin terbaik yaitu, IBA 0,1 mg/L dan BAP 0,05 mg/L. Kombinasi ini meningkatkan tinggi dan jumlah ruas Aquilaria beccariana dengan tinggi rata-rata sebesar 1,64 cm dan jumlah ruas rata-rata sebesar 6,40 ruas. Pada kombinasi dan taraf ini diduga mekanisme kerja IBA dan BAP paling efektif dibanding perlakuan yang lain. Sedangkan kombinasi IBA 0 mg/L dan BAP 0,03 mg/L memberikan respon terbaik terhadap peningkatan jumlah tunas dengan rata-rata sebanyak 1,91 tunas.Kata Kunci: Aquilaria beccariana, tunas, elongasi, auksin, sitokinin


2013 ◽  
Vol 5 (4) ◽  
pp. 518-523 ◽  
Author(s):  
Kishore Kumar CHIRUVELLA ◽  
Arifullah MOHAMMED ◽  
Rama Gopal GHANTA

Micropropagation has been advocated as one of the most viable biotechnological tool for ex situ conservation of rare, endangered endemic medicinal plants germplasm. Rapid clonal micropropagation protocol for large-scale multiplication of an endemic medicinal plant Soymida febrifuga (Meliaceae) was established from 15-day aseptic seedling cotyledonary node and shoot tip explants. High frequency of sprouting and shoot differentiation was observed from cotyledonary node explants compared to shoot tip, on Murashige and Skoog (MS) medium fortified with BA, KN, 2-iP and CM. Of the cytokinins used, BA (3.0 mgl-1) supported highest average number and maximum multiple shoot differentiation (16.6). In vitro proliferated shoots were multiplied rapidly by culturing nodal segments as microcuttings, further subcultured on the same media for elongation. Elongated shoots upon transfer to MS medium fortified with IBA showed rooting within two weeks of culture. Rooted plantlets were successfully hardened and 75% of rooted shoots successfully survived on establishment to the soil. Plants looked healthy with no visually detectable phenotypic variations. This protocol provides a successful and rapid technique that can be used for ex situ conservation minimizing the pressure on wild populations and contributes to the conservation of this endemic medicinally potent flora.


1992 ◽  
Vol 117 (2) ◽  
pp. 346-350 ◽  
Author(s):  
Isabel Arrillaga ◽  
Victoria Lerma ◽  
Juan Segura

A protocol for in vitro propagation in flowering ash (Fraxinus ornus L.) has been developed. Shoot apices or nodal segments from aseptically grown seedings or shoot apices from adult trees were used as initial explants. Highest shoot multiplication rates were obtained when the explants were cultured for 30 days in liquid Rugini induction medium supplemented with BA followed by 30 days on solidified Rugini multiplication medium without growth regulators. Regenerated shoots were rooted on Heller medium containing auxins alone or in combination with BA. Rooting percentages up to 71% (juvenile material) or 50% (adult material) were obtained in the presence of NAA and BA, and were not improved by treating the basal end of the shoots with concentrated NAA solutions. Following conventional procedures, regenerated plants were transferred to soil with more than 80% success. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA); 1-naphthaleneacetic acid (NAA).


2018 ◽  
Vol 13 (9) ◽  
pp. 1934578X1801300
Author(s):  
Daniele Fraternale ◽  
Marco Bruno ◽  
Luigi Rocchi ◽  
Antonella Amicucci ◽  
Chiara Ceccarini ◽  
...  

Roveja is a pea cultivar from the Sibillini Mountains (Umbria-Marche Apennine mountain range, Italy). Abandoned for long time, Roveja is nowadays cultivated only in few farms. The present work deals with the in vitro plant growth of this pea by tissue culture for shoots regeneration and ex situ conservation. Total polyphenols content evaluation, DPPH (2,2-diphenyl-1-picrylhydrazyl) antioxidant activity test and ORAC (Oxygen Radical Absorbance Capacity) assay were performed on the flour obtained from Roveja seeds and the results were compared to those collected by performing the same tests on the flour from two common green peas. The best shoots regeneration from embryonic axes was obtained in Murashige & Skoog (MS) media with combinations of 4.0 mg/L BA (6-Benzyladenine) plus 0.4 mg/L NAA (1-Naphthaleneacetic acid), and the best roots induction from the regenerated shoots was obtained in half strength MS media containing 1.0 mg/L IBA (3-Indolebutyric acid). The embryonic axes of Roveja seeds can be used as the starting material for in vitro regeneration of the plant; the antioxidant properties shown by this legume suggest further investigations to evaluate its nutritional/functional benefit.


HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2134-2137
Author(s):  
Flávia D. Pereira ◽  
José Eduardo B.P. Pinto ◽  
Luciana D.S. Rosado ◽  
Helen C.A. Rodrigues ◽  
Suzan K.V. Bertolucci ◽  
...  

The aim of the study was to develop a method for the in vitro propagation of Ananas erectifolius (Bromeliaceae), a fiber-rich Amazonian species. In vitro cultures were established from axillary buds of field-grown plants cultured on medium without plant growth regulators (PGRs). Stumps were excised from in vitro plantlets and incubated under dark conditions on medium supplemented with different combinations of 1-naphthaleneacetic acid (NAA), kinetin, and gibberellic acid (GA3). The most efficient induction of etiolated shoots occurred on explants cultured in the presence of NAA at 10.74 μm (T1 medium) or NAA at 5.37 μm + GA3 at 3 μm (T2 medium). Apical tips and nodal segments of the etiolated shoots were recultured under a 16-h photoperiod in medium without PGRs, and the effects of residual PGRs were evaluated by determining the numbers and lengths of plantlets that regenerated within 30 days. Residual PGRs exhibited no effect on the length of the regenerated plantlets but significantly affected the number of plantlets regenerated from nodal segments but not from apical tips. Nodal segments and apical tips derived from etiolated shoots produced, respectively, on T2 and T1 medium were most appropriate for plantlet regeneration. Nearly all (98%) regenerated plantlets formed roots when cultured in liquid medium without PGRs, and all plantlets survived acclimatization under greenhouse conditions. The stumps originating from etiolated shoots regenerated new etiolated shoots when recultured in the dark on medium without PGRs, thus providing a supply of new explants for plant regeneration.


2015 ◽  
pp. 83-98
Author(s):  
Marija Markovic ◽  
Mihailo Grbic ◽  
Matilda Djukic

The review of recent researches regarding the in vitro culture of 30 endangered Dianthus taxa is presented in this paper. Various in vitro protocols developed for selected rare and threatened Dianthus taxa are analysed in order to provide a useful synthesis of the data obtained with the main principles, techniques and recommendations for futher research and practice. The recapitulated data presented in this review can be used as a tool for the micropropagation of other endangered Dianthus taxa, enabling their propagation and obtaining a sufficient amount of plants for reintroduction. In addition, the obtained results represent the basis for ex situ conservation of the investigated taxa, especially for medium-term and long-term conservation (cryopreservation).


Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 934
Author(s):  
Chris O’Brien ◽  
Jayeni Hiti-Bandaralage ◽  
Raquel Folgado ◽  
Alice Hayward ◽  
Sean Lahmeyer ◽  
...  

Recent development and implementation of crop cryopreservation protocols has increased the capacity to maintain recalcitrant seeded germplasm collections via cryopreserved in vitro material. To preserve the greatest possible plant genetic resources globally for future food security and breeding programs, it is essential to integrate in situ and ex situ conservation methods into a cohesive conservation plan. In vitro storage using tissue culture and cryopreservation techniques offers promising complementary tools that can be used to promote this approach. These techniques can be employed for crops difficult or impossible to maintain in seed banks for long-term conservation. This includes woody perennial plants, recalcitrant seed crops or crops with no seeds at all and vegetatively or clonally propagated crops where seeds are not true-to-type. Many of the world’s most important crops for food, nutrition and livelihoods, are vegetatively propagated or have recalcitrant seeds. This review will look at ex situ conservation, namely field repositories and in vitro storage for some of these economically important crops, focusing on conservation strategies for avocado. To date, cultivar-specific multiplication protocols have been established for maintaining multiple avocado cultivars in tissue culture. Cryopreservation of avocado somatic embryos and somatic embryogenesis have been successful. In addition, a shoot-tip cryopreservation protocol has been developed for cryo-storage and regeneration of true-to-type clonal avocado plants.


Author(s):  
Asmaa Abdelsalam ◽  
Ehab Mahran ◽  
Kamal Chowdhury ◽  
Arezue Boroujerdi

Abstract Background Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. Results Twenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. Conclusions This study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.


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