scholarly journals Nutrient optimization for improved in vitro plant regeneration in Eclipta alba (L.) Hassk. and assessment of genetic fidelity using RAPD analysis

2015 ◽  
Vol 24 (2) ◽  
pp. 223-234
Author(s):  
Shruti Bardar ◽  
Varsha Khurana Kaul ◽  
Sumita Kachhwaha ◽  
SL Kothari
Keyword(s):  
Basal Medium ◽  
Genetic Fidelity ◽  
Ex Situ ◽  
Nutrient Level ◽  
Induction Medium ◽  
Eclipta Alba ◽  
Shoot Bud ◽  
Bud Induction ◽  
Regeneration Response

This study highlights the effect of different inorganic micronutrients like copper, cobalt, molybdenum, zinc, boron, iodine, iron and manganese in accelerating and amplifying in vitro shoot bud induction and proliferation of a medicinally important plant, Eclipta alba (L.) Hassk. Direct shoot bud induction was observed on MS fortified with Kn (2 mg/l). However, maximum number of shoots was achieved when GA3, 0.5 mg/l was added to induction medium along with 1?M copper sulphate (ten times the normal MS level). Optimization of nutrient level in the basal medium promoted maximum regeneration response from both shoot tips and nodal explants. Elongated shoots were rooted in MS supplemented with IBA, 1.0 mg/l. Healthy, green plantlets with well developed roots, flowered normally in the field. Genetic stability of micropropagated plantlets was evaluated using RAPD markers. The amplification products were monomorphic in micropropagated plantlets and similar to those of mother plant revealing the genetic uniformity of plantlets. The regeneration protocol is highly efficient and reproducible so would be useful for mass multiplication, ex situ conservation and genetic transformation of E. alba (L.) Hassk.Plant Tissue Cult. & Biotech. 24(2): 223-234, 2014 (December)

scholarly journals In vitro callus induction in inflorescence segments of medicinally important endangered plant Rauwolfia serpentina (L.) Benth. ex Kurz – a step towards ex situ conservation

2018 ◽  
Vol 7 (2) ◽  
pp. 1986
Author(s):  
Saranjeet Kaur
Keyword(s):  
Acetic Acid ◽  
Ex Situ Conservation ◽  
Ex Situ ◽  
Naphthalene Acetic Acid ◽  
Endangered Plant ◽  
Shoot Bud ◽  
Rauwolfia Serpentina ◽  
Entire Plant ◽  
Regeneration Response

The regenerative competence of inflorescence segments of Rauwolfia serpentina was assessed in M (Mitra et al., 1976) medium and its combinations with growth adjuncts such as cytokinins [N6-benzyladenine (6-BA - 0.5, 1.0 mg l-1), furfurylaminopurine (Kn-0.5, 1.0mg l-1) and auxin [α-naphthalene acetic acid (NAA - 0.5, 1.0 mg l-1)]. Inflorescence segments (1.00 cm in length) were used as explants. The regeneration response was obligatory to the use of growth regulators in the medium. Cytokinins favoured shoot bud mediated pathway of plantlet development and BAP favoured early shooting. The explants callused in NAA enriched medium. The callus was best maintained in NAA (0.5 mg l-1) through repeated sub-culturing. The entire plant of R. serpentina is the target of massive commercial collections resulting into shrinking habitats. Present study is the first ever report on the successful use of inflorescence segments for ex situ conservation of R. serpentina.

scholarly journals In vitro Regeneration of Blepharispermum subsessile DC: An Endangered Medicinal Plant of Odisha, India using Cotyledon Explants

2016 ◽  
Vol 26 (2) ◽  
pp. 255-266 ◽  
Author(s):  
Pranati Nayak ◽  
Kalidass C
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Garden Soil ◽  
Induction Medium ◽  
Root Induction ◽  
Shoot Buds ◽  
Shoot Bud ◽  
Cotyledon Explants ◽  
Bud Induction

Multiple shoots were induced on cotyledon explants of in vitro grown seedlings of Blepharispermum subsessile DC, cultured on MS medium supplemented with various combinations and concentrations of BAP, IBA and GA3. The highest regenerative response was observed on medium containing 2.5 mg/l BAP where shoot buds initiated after 12 days of inoculation and about 32 shoots were produced in 30 days time. Addition of GA3 played a key role in leaf expansion and elongation of shoot buds. Addition of the auxin IBA to the induction medium resulted in more callus proliferation rather than shoot bud induction. The elongated shoots were transferred to root induction medium consisting of half strength MS supplemented with IAA, NAA and IBA. Highest rooting response (90%) was recorded in ½ MS supplemented with 1.0 mg/l IAA. Acclimatized plants were maintained in polybags with garden soil for future reintroduction program to their natural habitat.Plant Tissue Cult. & Biotech. 26(2): 255-266, 2016 (December)

scholarly journals Glutamine enhances competence for organogenesis in pineapple leaves cultivated in vitro

2005 ◽  
Vol 17 (4) ◽  
pp. 383-389 ◽  
Author(s):  
Regina M. Hamasaki ◽  
Eduardo Purgatto ◽  
Helenice Mercier
Keyword(s):  
Direct Organogenesis ◽  
Induction Medium ◽  
Induction Phase ◽  
Regeneration Rate ◽  
Shoot Buds ◽  
Shoot Bud ◽  
Bud Induction ◽  
Shoot Induction Medium ◽  
Pineapple Leaves

Leaf bases of pineapple cultured on a shoot induction medium (SIM) produced protuberances followed by shoot-buds via direct organogenesis at a frequency of 46 %. When 8 mM glutamine (gln) was a supplement to SIM (SIM8gln), the regeneration rate increased to 70 %, thus suggesting that 8mM gln increased explant competence for organogenesis. Besides this, shoot vigor was strongly enhanced in SIM8gln. Other gln concentrations (16 or 32 mM) evoked a lower frequency of shoot-bud induction and number of regenerated shoots per explant when compared to SIM8gln. In this study, it was defined that explant organogenic commitment to form shoot-buds occurred in the first 7 days of culture on SIM8gln. Thereafter, endogenous indole-3-acetic acid (IAA) and cytokinin (4 types) measurements were carried out during this period, that is, during the induction phase of shoot-bud formation. The IAA content increased greatly until the 5th day in the leaf bases cultured on SIM8gln. No such change in IAA concentration was observed in the explants cultivated on SIM or in the presence of the highest gln concentration (32 mM), this being inhibitory to the organogenic process. The only natural cytokinin detected was isopentenyladenine. An increase of 50 % in the level of this phytohormone occurred in leaf bases cultured on SIM8gln at the 5th day, when compared to SIM or of 170% compared to SIM32gln. These results suggest that 8 mM gln favorably influenced the organogenic process through changes in IAA and iP concentrations in pineapple leaves.

scholarly journals Metabolic profiling, in vitro propagation, and genetic assessment of the endangered rare plant Anarrhinum pubescens

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Asmaa Abdelsalam ◽  
Ehab Mahran ◽  
Kamal Chowdhury ◽  
Arezue Boroujerdi
Keyword(s):  
In Vitro Propagation ◽  
Genetic Fidelity ◽  
Ex Situ ◽  
Wild Plant ◽  
Rare Plant ◽  
Chemical Profiling ◽  
Nodal Cutting ◽  
Regenerated Plants ◽  
Plant Shoots

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.

scholarly journals In vitro organogenesis and plant regeneration of Passiflora xishuangbannaensis, a species with extremely small populations

2021 ◽  
Author(s):  
Yuan-yuan Meng ◽  
Shi-jie Song ◽  
Sven Landrein
Keyword(s):  
Plant Regeneration ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Basal Medium ◽  
Ex Situ ◽  
Naphthaleneacetic Acid ◽  
South American ◽  
South American Species

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species

scholarly journals In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media

2020 ◽  
Vol 21 (11) ◽  
Author(s):  
Yupi ISNAINI ◽  
Titien Ngatinem Praptosuwiryo
Keyword(s):  
Spore Germination ◽  
Culture Media ◽  
Basal Medium ◽  
Modern Medicine ◽  
Ex Situ ◽  
Naphthalene Acetic Acid ◽  
Gametophyte Development ◽  
In The Wild ◽  
Rare And Endangered

Abstract. Isnaini Y, Praptosuwiryo TNg. 2020. In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media. Biodiversitas 21: 5373-5381. Cibotium barometz (L.) J. Sm. is known as the golden chicken fern and included in Appendix II of CITES. It is an important export commodity for traditional and modern medicine. Globally, populations of this species are under significant pressure due to overexploitation in the wild. In vitro culture is one of the technologies used for ex-situ propagation and conservation of rare and endangered ferns and lycophytes. This study’s objectives were: (i) to observe in vitro spore germination and early gametophyte development of C. barometz, and (ii) to determine the best culture medium for rapid spore germination and early development of the gametophytes. The sterilized spores were sown in half-strength Murashige & Skoog (½MS) basal medium supplemented with combinations of 6-Benzylaminopurine (BAP) and α-Naphthalene acetic acid (NAA). A factorial combination of four BAP concentrations (0, 2, 4, and 6 mg L-1) with four concentrations of NAA (0; 0.01; 0.03 and 0.05 mg L-1) created 16 treatments replicated in a Completely Randomized Design. Spore germination of C. barometz was observed to be Vittaria-type, and its prothallial development was Drynaria-type. Spore germination started 7-14 days after sowing. Young heart-shape gametophytes consisting of 110-240 cells were formed in 45-61 days after sowing. The two best spore culture media for rapid spore germination and development of C. barometz gametophytes were ½ MS with or without 2 mg L-1 BAP.

scholarly journals In Vitro Propagation of an Endangered Helianthus verticillatus by Axillary Bud Proliferation

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 712
Author(s):  
Marzena Nowakowska ◽  
Žaklina Pavlović ◽  
Marcin Nowicki ◽  
Sarah L. Boggess ◽  
Robert N. Trigiano
Keyword(s):  
Endangered Species ◽  
Shoot Proliferation ◽  
Genetic Fidelity ◽  
Induction Medium ◽  
Axillary Shoot ◽  
Axillary Shoot Proliferation ◽  
Regenerated Plants ◽  
Micropropagated Plants ◽  
Ssrs Markers

Helianthus verticillatus (Asteraceae), whorled sunflower, is a perennial species restricted to a few locations in the Southeastern United States. Habitat loss has caused H. verticillatus to become rare, and since 2014, it has been federally listed as an endangered species. As a part of the recovery plan for the restoration and protection of H. verticillatus, an efficient micropropagation protocol based on axillary shoot proliferation was developed. Various concentrations of 6-benzylaminopurine (BAP; 0 to 4.44 µM) were examined for their morphogenetic potential in the regeneration of six genotypes of H. verticillatus from the nodal explants derived from greenhouse-grown plants. Both the BAP concentration and genotype had significant effects on the regeneration capacity of H. verticillatus. Although the induced buds were observed on ½-strength Murashige and Skoog medium without plant growth regulators, a higher rate of induction and bud development were achieved on media with either 0.88 or 2.22 µM BAP, regardless of the genotype. Successful rooting of the induced shoots was achieved within four weeks after the transfer from the induction medium to the fresh ½-strength MS medium, but the rooting efficiency was dependent on the plant’s genetic background. Regenerated plantlets, with well-developed shoots and roots, were acclimatized successfully to greenhouse conditions with a 97% survival rate. Simple sequence repeats (SSRs) markers were employed to assess the genetic uniformity of the micropropagated plants of H. verticillatus. No extraneous bands were detected between regenerants and their respective donor plants, confirming the genetic fidelity and stability of regenerated plants. To our knowledge, the protocol developed in this study is the first such report for this endangered species.

scholarly journals Confirmation of “pre-plasmolysis mediated ex-osmosis hypothesis” to obtain shoot bud morphogenesis in Catharanthus roseus

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Vyoma Mistry ◽  
Abhishek Sharma ◽  
Ajay Kumar Mathur
Keyword(s):  
Genetic Engineering ◽  
Catharanthus Roseus ◽  
Cultured Cells ◽  
De Novo ◽  
Leaf Explants ◽  
Direct Regeneration ◽  
Engineering Process ◽  
Shoot Bud ◽  
Regeneration Response

AbstractThe antineoplastic herb, Catharanthus roseus is a classified high-value low-volume medicinal herb which is in global attention of scientific research for modulation of its monoterpenoid indole alkaloids (MIA) pathway through genetic engineering. These secondary metabolites are generally stored in specific types of structures/compartments due to their cytotoxic nature and designated roles in plant defense response. However, their presence can hinder the genetic engineering process used to develop transgenic plants through de novo morphogenesis and regeneration of plants from cultured cells/tissues and hence, it always remained a critical impediment in transgenic research in C. roseus. The pre-plasmolysis treatment of leaf explants can help to tackle the recalcitrant nature of leaf explant and can support the direct regeneration response by ex-osmosis that minimizes the concentration of alkaloids. Therefore, this study was performed to chase the effect of osmotic conditions on recalcitrant leaves of C. roseus engaged in vitro plant regeneration and hypothesis of alkaloids ex-osmosis is confirmed by HPLC analysis.

scholarly journals Plant Regeneration from in Vitro Culture of Embryonic Axis Explants in Common and Tepary Beans

1992 ◽  
Vol 117 (2) ◽  
pp. 332-336 ◽  
Author(s):  
Mohamed F. Mohamed ◽  
Paul E. Read ◽  
Dermot P. Coyne
Keyword(s):  
Plant Regeneration ◽  
Common Bean ◽  
Basal Medium ◽  
Continuous Light ◽  
Embryonic Axis ◽  
Naphthaleneacetic Acid ◽  
Tepary Bean ◽  
Bud Induction ◽  
Multiple Buds

A new in vitro protocol was developed for multiple bud induction and plant regeneration from embryonic axis explants of four common bean (Phaseolus vulgaris L.) and two tepary bean (P. acutifolius A. Gray) lines. The explants were prepared from two embryo sizes, 3 to 4 mm and 5 to 7 mm, corresponding to pods collected after 15 and 25 days from flowering, respectively. The embryonic axis was cultured on Gamborg's B5 basal medium with 0, 5, 10, or 20 μm BA in combinations with 0, 1, or 2 μm NAA. The cultures were maintained at 24 to 25C under continuous light or incubated in darkness for 2 weeks followed by continuous light before transfer to the secondary B5 medium (0 or 2 μm BA or 2 μm BA plus 4 μm GA3). Adventitious roots or a single shoot with roots formed on the explants cultured on media without plant growth regulators. Multiple buds were induced on all BA media, but more were produced with 5 or 10 μm for most lines. Dark incubation greatly enhanced multiple bud initiation. Shoot buds were not produced on media containing NAA alone or in combinations with BA. On the secondary medium, six to eight shoots per explant for common bean and up to 20 shoots per explant from tepary bean were observed after 3 weeks. Mature, fertile plants were produced from these shoots. Chemical names used: benzyladenine (BA); 1-naphthaleneacetic acid (NAA); gibberellic acid (GA3).

scholarly journals HISTOLOGICAL EXAMINATION OF IN VITRO ADVENTITIOUS BUD DEVELOPMENT ON HYPOCOTYLS OF FRASER FIR

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1102a-1102
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson
Keyword(s):  
Growth Regulators ◽  
Induction Medium ◽  
Adventitious Bud ◽  
Naphthaleneacetic Acid ◽  
Abies Fraseri ◽  
Fraser Fir ◽  
Cell Clusters ◽  
Cell Divisions ◽  
Bud Induction

Hypocotyls of Fraser fir (Abies fraseri (Pursh) Poir.) were excised from seeds germination 9 days and placed on bud induction medium containing 10 mg/liter benzyladenine (BA) and 0.01 mg/liter naphthaleneacetic acid (NAA) or medium without growth regulators. After 3 days on medium containing growth regulators, cell divisions were localized in epidermal and subepidermal layers of the hypocotyl while similar cell divisions were not observed in control-treated hypocotyls. Cell clusters consisting of two to five cells were present after 7 days in hypocotyls placed on bud induction medium. In control-treated hypocotyls, stomata continued to develop and cells within the cortex became vacuolated during the first 2 weeks in culture. All hypocotyls were transferred to secondary medium after 3 weeks. Cell clusters continued to enlarge into meristemoids in hypocotyls initially placed on bud induction medium. Gradually, meristemoids developed into buds and cataphylls were observed covering bud meristems.

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