scholarly journals Preparation of Synthetic Seeds of Citrus jambhiri Using in vitro Regenerated Multiple Plantlets

Author(s):  
Priyanka Sharma ◽  
Bidhan Roy

Biotechnological tools are useful for true-to-type propagation. Shoot tips encapsulation is potential for plant development from pre-existing meristematic tissue. MS medium fortified with 1 and 2 mg/L of BAP (6-bezylaminopurine) was found to be suitable for in vitro mass-multiplication of plantlets (10.18 and 13.05 plantlets/explant, respectively) of Citrus jambhiri from nodal segments. Nodal segments were more appropriate than the shoot tips for in vitro multiplication of plantlets. Synthetic seeds were prepared using 2.5% sodium alginate dropping into 3.0% CaCl2 solution. Maximum germination was recorded when beaded shoot tips were cultured on MS medium fortified with 1 and 2 mg/L of BAP (96.67 and 100.00%, respectively). However, the germination of synthetic seeds was found to be comparatively high than the earlier findings. The results support the use of encapsulated unipolar explants for synthetic seed preparation. These type of capsules could be useful in exchange of sterile material between laboratories, germplasm conservation and direct plant propagation.

2011 ◽  
Vol 3 (1) ◽  
pp. 29-33 ◽  
Author(s):  
K. K. Singh ◽  
Bhusan Gurung

A method has been developed for plant regeneration from alginate-encapsulated nodal segments of Rhododendron dalhousiae. Shoot tips collected from in vitro proliferated shoots were used for synthetic seed production. For encapsulation, nodal segments were mixed with MS medium supplemented with 3% sodium alginate and incubated with calcium chloride (60 mM). The maximum frequency (69%) of conversion of encapsulated shoot tips into plantlets was achieved on MS medium containing 25 μM 2-isopentenyladenine (2iP) along with additive such as, 100 mg L-l polyvinyl pyrrolidone (PVP), 100 mg L -l ascorbic acid, 10 mg L-l citric acid. The presence of 2iP (25 μM) with IAA (0.6 μM) improved re-generation. Amongst the two gelling agents used higher shoot proliferation as well as better growth were observed in cultures grown on Agar in comparison to Phytagel medium. Encapsulated nodal segments stored at 4°C for 25 days also showed successful conversion, followed by development into complete plantlets when returned to regeneration medium. Liquid medium was superior over solid medium for root formation and growth. IBA (1.0 μM) was more effective than other auxins for root induction. Plantlets with developed shoot and roots were hardened off to survive ex vitro conditions and successfully established in greenhouse. Possibility of direct sowing of synthetic seeds in the soil was also examined.


Medicinal plants are one of the most vital natural resources, but many of them are currently endangered due to habitat loss. Consequently, it is critical to emphasize the importance of using micropropagation techniques for mass propagation of plantlets on a commercial scale, in addition to germplasm conservation and distribution. Nodal explants and shoot tips were expunged from 15 days of the explant by aseptic seedlings, an effective, quick, and better in vitro plant regeneration procedure for Vitex negundo L. has been developed. The recent study was considered to develop an in vitro procedure for the regeneration of V. negundo L., a traditional medicinal plant. Nodal segments and shoot tips were cultivated on MS medium enhanced with numerous plant growth regulators. For multiple shoots and root regeneration, various cytokinins were examined. 6-benzyl-aminopurin (BAP), kinetin (Kin), and 1H-indole-3-butanoic acid (IBA) were all tested as a supplement to Murashige and Skoog (MS) medium including auxin phytohormone, such as Indole acetic acid (IAA) and 1-naphthaleneacetic acid (NAA). The furthermost effective surface sterilization treatment for explants of V. negundo has been found 0.1% HgCl2 for 8 minutes. In all treatments, multiple shoots were collected from shoot tips and nodal segments. In MS media added with 2.0mg/l BAP, the most shoots were seen in V. negundo. Furthermore, V. negundo regeneration shoots rooted effectively in half MS containing 1.0 mg/l IBA. Finally, proliferated plantlets were effectively adapted in soil, where they grew normally without morphological anomalies and had a survival rate of 92 percent.


Author(s):  
S. Parvin ◽  
M. Kausar ◽  
M. Enamul Haque ◽  
M. Khalekuzzaman ◽  
B. Sikdar ◽  
...  

A rapid and efficient protocol is outlined for in vitro propagation of muskmelon(Cucumis melo L.) Shoot tips, nodal segments and cotyledonary nodes from invitro grown seedlings were used as explants. The explants were inoculated on MS medium fortified with different combinations and concentrations of growthregulators viz., BAP, NAA, GA3 and IBA for multiple shoot regeneration.Effective result was found on MS medium supplemented with 2.0 mg/l BAP, inwhich 90% and 70% cultures induced multiple shoots from nodal segments andshoot tip explants, respectively. Whereas, 70% cultures of cotyledonary nodeswere found to induced shoots on MS medium with 1.5 mg/l BAP + 0.1 mg/l GA3. In vitro regenerated shoots were subcultured on half strength MS mediumsupplemented with different concentrations of IBA and NAA for successful rootinduction and the effective result (up to 70%) was found in medium with 1 mg/lIBA. Well rooted in vitro grown plantlets were acclimatized in sandy soil, whereas 70% plantlets survived


Author(s):  
D. L. C. K. Fonseka ◽  
W. W. U. I. Wickramaarachchi ◽  
R. P. S. Madushani

The black-oil tree (Celastrus paniculatus Willd) is a highly valued medicinal plant species belong to the Celastraceae family, known as Jyothishmathi in Ayurveda and Duhundu in Sri Lanka and grows as a perennial vine. It is an endangered medicinal plant species recorded in the red list of endangered fauna and flora of Sri Lanka in 1999. The seed oil of Celastrus paniculatus contains sesquiterpene alkaloids namely; celapagine, celapanigine, celapanine and celastrol, used in traditional system of medicine for various disorders and because of its high pharmaceutical value, plants are over exploited in natural habitats. Owing to poor seed germination and lack of successful vegetative propagation methods, domestication and commercial planting of this important medicinal plant species to meet the demand seems impossible. Therefore, it is of high importance to develop a reliable and efficient in vitro propagation to produce black oil plants for commercial use. In this study, it was attempted to produce synthetic seeds of Celestrus paniculatus via in vitro multiple shoot proliferation. Nodal segment explants were collected from freshly emerged age of sprouts, surface sterilized and cultured in Murashige and Skoog medium supplemented with different 6-benzylaminopurine (BAP) and Thidiazuron (TDZ) concentrations for shoot induction. The highest soot proliferation rate; 25 shoot tips/explant were observed with 0.1 mg/L TDZ. Induced shoot tips were used for synthetic seed production after encapsulating with BAP and a-naphthalene acetic (NAA) enriched sodium alginate. Shoot tip encapsulated beads produced with 4% sodium alginate were firm, clear, round and uniform in size and easy to handle. The influence of growth regulators (BAP and NAA) and storage period on the germination of encapsulated shoot tips was studied to evaluate the success of encapsulated shoot tips as a propagule. The beads germinated with 2 mg/L BAP and 0.2 mg/L NAA provided 80% in vitro germination percentage. Shoot tips of synthetic seeds remained green and healthy after storage at 5°C for a period of 8 weeks. Current findings suggest that encapsulated micro shoots (synthetic seeds) could be produced successfully, as the first step in domestication and conservation of Celastrus paniculatus. Further studies required on rooting of micro shoots, acclimatization and transferring of plantlets produced from synthetic seeds to in vivo conditions for domestication and conservation purposes.


Forests ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 463
Author(s):  
Qiansheng Li ◽  
Mengmeng Gu ◽  
Min Deng

Quercus aliena Blume, also known as the oriental white oak, is a widespread species in temperate forests of East Asia with significant ecological and economical importance. Establishing an efficient vegetative propagation system is important for its germplasm conservation and breeding program. Protocols of micropropagation from shoot tips and nodal segments were investigated in order to produce uniform high-quality seedlings. Nodal segments from 18 month old seedlings were used as explants to initiate the aseptic culture. The highest bud proliferation was achieved by subculturing the explants on 1/2 strength woody plant medium (WPM) with 2.0 mg·L−1 BA. WPM with 0.5 mg·L−1 BA and 0.05 mg·L−1 IBA was the best medium for subculture to obtain the vigorous regenerated shoots in this experiment. Nodal segments without shoot tips had a higher adventitious bud proliferation rate than those with shoot tips. The highest rate (41.5%) of rooting in vitro was induced by using WPM with 1.0 mg·L−1 IBA and 5 g·L−1 activated charcoal. Ex vitro rooting by dipping the proliferated shoots with 500 mg·L−1 IBA solution, then transplanting directly to potting mix with 50% peat and 50% horticultural perlite fostered the highest rooting percentage and survival rate of the plantlets.


2020 ◽  
Vol 100 (2) ◽  
pp. 202-208
Author(s):  
Mengting Wang ◽  
Guiliang Zhang ◽  
Peiyao Xin ◽  
Yun Liu ◽  
Bin Li ◽  
...  

Camellia fascicularis is an endangered evergreen ornamental plant with pale yellow flowers. An efficient and reproducible in vitro regeneration method is required for its large-scale propagation and germplasm conservation. In this study, one axillary bud per nodal stem was obtained from C. fascicularis cultured on Murashige & Skoog (MS) medium containing 0.1 mg L−1 indole-3-acetic acid (IAA) combined with 1.0 mg L−1 6-benzylaminopurine (BA). Axillary buds from the stem segments were transferred to modified woody plant medium (WPM) supplemented with 3.0 mg L−1 BA in combination with 0.3 mg L−1 IAA for multiplication, thereby resulting in a high shoot multiplication rate of 6.8. Multiple shoots were divided into nodal stems and shoot tips and were induced to root. The shoot tips were induced to root by culturing on one-half MS medium supplemented with 2.0 mg L−1 indole-3-butyric acid (IBA) in combination with 0.3 mg L−1 α-naphthalene acetic acid (NAA), which resulted in 76.0% rooting efficiency with 2.3 roots per shoot. The optimal hormone ratio for inducing rooting of nodal stems was 1.0 mg L−1 IBA in combination with 2.0 mg L−1 NAA, which resulted in 72.7% rooting efficiency with 1.7 roots per nodal stem. These two rooted plantlets were successfully acclimatized and established in a greenhouse.


1970 ◽  
Vol 18 ◽  
pp. 140-145 ◽  
Author(s):  
Md Abu Hena Mostofa Jamal ◽  
ANM Rubaiyath-Bin Rahman ◽  
Dipak Kumar Paul ◽  
Md Rezuanul Islam

Context: It is necessary to focus on the importance of adopting micropropagation technique for mass propagation of the plantlets in commercial scale as well as conservation and distribution of germplasm. Objective: The present investigation has been designed with a view to establishing protocol of in vitro regeneration of medicinal plant species i,e., Vitex nigundo L (Verbenaceae). Materials and Methods: Shoot tips and nodal segments were used for multiple shoot induction. All explants were cultured on MS medium supplemented with various plant growth regulators. HgCl2 was used as surface sterilizing agent. For in vitro rooting, individual shoots (3-4 cm) were cut from the proliferated shoot cultures and implanted on half and full strength of MS with different concentrations and combinations of NAA and IAA. The cultures were incubated for 16 h photoperiod at 25 ± 2ºC under a fluorescent light. Visual observation of culture was made every week. Data on shoot induction and proliferation and root induction were recorded after three weeks of inoculation and used for calculation. For each treatment 15 explants were used and all the treatments were repeated thrice. Established plantlets were transplanted in earthen pots under natural conditions and the survival rate was recorded. Results: The most effective surface sterilization treatment has been found 0.1 % HgCl2 for 7 minutes. Highest number of shoot was observed in MS medium supplemented with 3.0 mg/ BAP. It was rooted well in full MS containing 2.0 mg/l IAA. The survival rate was 85 % and propagated plantlets were successfully acclimatized in soil. Conclusion: It was observed that shoot tips are more responsive for micropropagation of Vitex nigundo L . Thus the fruitful utilization of rapid clonal propagation, germplasm conservation and distribution of Vitex nigundo, important medicinal plant of Bangladesh, is possible. Keywords: Vitex nigundo; Medicinal plant; Shoot induction; Micropropagation; Regeneration. DOI: http://dx.doi.org/10.3329/jbs.v18i0.8790 JBS 2010; 18(0): 140-145


Author(s):  
Mohmad Amin ◽  
Abdul Mujeeb

The study was carried out for callus induction and synthetic seed development from the shoot tips of Draceana sanderiana sander ex Mast. The shoot tips were subjected to different concentrations (0.25, 0.5 &1.0 mg/l) of 2,4-D on MS medium. The research findings revealed that the 2,4-D at concentrations of 0.25 mg/l was more suited for the profuse callus formation. The friable and light yellow callus was induced within 2 weeks of culture at 0.25 mg/l of 2,4-D on MS medium as compared to the other two concentrations of 2,4-D i.e.; 0.5 and 1.0 mg/l. Similarly the effect of sodium alginate and calcium chloride percentage on synthetic seed formation was observed, it was found that somatic embryos formed from shoot tips via callus kept in 2.5% sodium alginate and 100 milli molar CaCl2 produced synthetic seeds with firm spherical beads. The study leads to the formation of synthetic seeds of Draceana sanderiana which can be used for the conservation of germplasm through cryopreservation and the micro propagation of the said plant species.


2017 ◽  
Vol 45 (1) ◽  
pp. 208-214 ◽  
Author(s):  
Ewelina KWAŚNIEWSKA ◽  
Ewa DZIEDZIC ◽  
Bożena PAWŁOWSKA

Cryopreservation is an useful technique for long-term conservation that requires minimal space and maintenance. Germplasm protection of Rosa is important to preserve genetic diversity, to store material for breeding and to expand new research. This study was conducted to develop a droplet vitrification cryopreservation and micropropagation of Rosa pomifera cv. ‘Karpatia’, whose large hypanthia are characterized by remarkable pro-health properties. Culture in vitro was stabilized and shoot tips collected from dormant buds served as initial explants. The multiplication of shoots was carried out on MS medium containing benzyladenine. For the droplet vitrification cryopreservation, shoot tips from in vitro cultures were used: small with exposed meristem, and large with a meristem covered with leaves, as well as shoot tips from in situ plants, which were collected in winter. Treatment time with plant vitrification solution (PVS2) was also tested (10-30 minutes). From in vitro culture, 32-41% small explants with exposed meristem survived, but they regenerated at a very low level. The best cryostorage results were obtained for shoot tips from dormant buds and a 20-minute PVS2 treatment: the survival was 84% and regeneration 72%. During the post-freezing regeneration multiplication index was 2.4 shoots per one multiplication cycle, after cryopreservation and in the control. On half MS medium without growth regulators, 97-99% of shoots rooted, and all rooted plants have adapted to ex vitro conditions and were planted into the soil. Biometric analyses during shoot multiplication, rooting and acclimatization stages did not reveal any changes compared to the non-cryopreserved samples.


2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Hamidou F. Sakhanokho ◽  
Cecil T. Pounders ◽  
Eugene K. Blythe

Synthetic seeds were formed from shoot tips of twoin vitrogrownBegoniacultivars using 3% sodium alginate in Murashige and Skoog medium (MS) salt solution as the gel matrix and 100 mM calcium chloride for complexation. Synthetic seed formation was achieved by releasing the sodium alginate/explant combination into 100 mM calcium chloride (CaCl2·H2O) solution for 30 or 45 min. Both control and encapsulated shoots were transferred into sterile Petri dishes and stored at 4°C or 22°C for 0, 2, 4, 6, or 8 weeks. Conversion of synthetic seeds into plantlets for both storage environments was assessed in MS medium or peat-based substrate. No significant difference was found between the 30 and 45 min CaCl2·H2O treatments or the two cultivars. Encapsulation of explants improved survival rate over time irrespective of the medium type or storage environment. Survival rates of 88, 53, 28, and 11% for encapsulated microshoots versus 73, 13, 0, and 0% for control explants were achieved in microshoots stored for 2, 4, 6, and 8 weeks, respectively. The best results were obtained when synthetic seeds were stored at 4°C and germinated on MS medium. Regenerated plantlets were successfully established in potting soil.


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