scholarly journals Effect of Nitrogen on in Vitro Propagation of Endangered Medicinal Plant: Swertia Chirayita Roxb. Ex Flaming

2021 ◽  
Vol 11 (2) ◽  
pp. 3513-3522
Keyword(s):  
Average Length ◽  
Inorganic Nitrogen ◽  
Nitrogen Sources ◽  
Multiple Shoots ◽  
Garden Soil ◽  
Equal Proportion ◽  
Ms Medium ◽  
Root Explants ◽  
Swertia Chirayita

The effect of nitrogen was investigated on the organogenesis of Swertia chirayita (Gentianaceae) to overcome the challenges related to its cultivation. The best callogenic response was observed on root explants inoculated onto MS medium supplemented with BAP (2.0 mg/l) along with 2,4-D (0.5 mg/l) after 35 days of culture. Subsequent transfer of callus for multiplication on the same media composition under complete darkness presents the best results in terms of callus multiplication. Callogenic cultures were subculture onto modified MS medium supplemented with inorganic nitrogen sources, i.e., NH4NO3 (14-56N/l), KNO3 (100-400N/l) with BAP (3.0 mg/l) were observed. Organogenic response (52%) was observed after 8-12 weeks of culturing. The maximum number of the shoot was recorded on MS medium with NH4NO3 (28 N/l), KNO3 (300N/l) with BAP (3.0 mg/l). Moreover, 90% of them were able to regrow when sub-cultured on the same media. Sixteen weeks old multiple shoots were subcultured on MS medium supplemented with different auxins. IAA was proved to be the best hormone rooting purpose. However, the best rooting response regarding the number of roots and an average length of roots was obtained at IAA (1.0 mg/l). Survival of 90% was achieved when rooted plantlets were successfully established in substrate containing sand, vermicompost, and garden soil in equal proportion for hardening and acclimatized.

scholarly journals Establishment of in vitro regeneration protocol for Adhatoda vasica Nees

2016 ◽  
Vol 51 (1) ◽  
pp. 75-80 ◽  
Author(s):  
S Khan ◽  
S Akter ◽  
A Habib ◽  
TA Banu ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Adhatoda Vasica

An in vitro regeneration protocol of Adhatoda vasica has been developed using excised nodal segments and juvenile leaves for multiple shoots regeneration directly or through callus induction. Explants were cultured on MS medium with different concentrations of IAA, NAA, BAP, GA3 and Kn singly or in combinations. MS medium supplemented with BAP (10.0 mg/l) was found best for multiple shoot formation, in which 93.33% explants produced multiple shoots. After two months, maximum number of multiple shoots were 10.6 ± 1.82, highest length of plantlets was 5.2 ± 2.20 cm. 100% calli formation were observed on MS medium supplemented with IAA (0.05 mg/l) + NAA (0.05 mg/l) + BAP (1.0 mg/l). Callus initiation started after 14 days and gave light green colored callus. Best callus mediated shoot regeneration was found on MS+10.0 mg/l BAP medium. Root induction of in vitro raised shoots was best on ½ MS + IBA (1.0 mg/l). Well rooted plantlets were transferred to plastic pots containing garden soil and compost in a ratio of 2:1 for hardening. The ultimate survival rate under natural condition was about 80%.Bangladesh J. Sci. Ind. Res. 51(1), 75-80, 2016

scholarly journals Chromosome stability of in vitro propagated Cucurbita cultivars

2019 ◽  
Vol 3 (3) ◽  
pp. 25
Author(s):  
Buse Dursun ◽  
Ahu Altınkut Uncuoğlu ◽  
Yıldız Aydin
Keyword(s):  
Chromosome Number ◽  
Cucurbita Pepo ◽  
Multiple Shoots ◽  
Cotyledonary Node ◽  
Garden Soil ◽  
Ms Medium ◽  
Ploidy Levels ◽  
Root Regeneration ◽  
Cucurbitaceae Family

Cucurbita pepo L. which is a member of Cucurbitaceae family, is a one-year plant with herbaceous stems, broad leaves and superficial scattered roots. Monoic flower structure in the Cucurbitaceae family and the differences in the maturing time of male and female organs in flowers cause an increase in the foreign fertilization rate. Therefore, there may be positive or negative changes in the existing characteristics of the species. Micro-propagation method can be performed in pumpkin species for clonal propagation, but genetic stability after tissue culture is an important consideration. Chromosome number and morphology are primary cytogenetic parameters that must remain stable after in vitro propagation. We analyzed plants of different hybrid pumpkin genotypes cultivated in our country (Ardendo, Angelina, Torpido, Roni, Sena Hanım) cytogenetically in order to determinate their stability level. Cotyledon nodes, nodes, shoot apex, hypocotyl and internode explants were prepared from the 4-week old C. pepo seedlings by making a horizontal slice through the hypocotyl region. Cotyledonary node explants produced multiple shoots and callus regeneration in MS medium in the presence of N6-benzylamino-purine BA 1 mg/L in Torpido genotype. Elongated shoots were excised from shoot clumps and transferred to rooting medium. The best root generation was obtained from Torpido and the most successful explant type for root regeneration was determined in cotyledon nodes as in shoot regeneration and MS medium without plant growth regulator showed the best root regeneration. The rooted plants were hardened in small pot containing standardized garden soil, well developed plant transferred to greenhouse. The chromosome number and karyotype analysis were determined in control and in vitro propagated Cucurbita pepo L. plants and ploidy levels were confirmed to be 2n = 40.

scholarly journals In vitro propagation and cytological analysis of Sophora mollis Royle: an endangered medicinal shrub

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Aakriti Bhandari ◽  
Harminder Singh ◽  
Amber Srivastava ◽  
Puneet Kumar ◽  
G. S. Panwar ◽  
...  
Keyword(s):  
Chromosome Number ◽  
Average Length ◽  
Germplasm Conservation ◽  
Basic Chromosome Number ◽  
Shoot Tip ◽  
Ex Situ ◽  
Ms Medium ◽  
Basic Chromosome ◽  
Shoot Tip Explants

Abstract Background Sophora mollis Royle (family Fabaceae, subfamily-Papilionaceae) is a multipurpose legume distributed in plains and foothills of the North-West Himalaya to Nepal and is facing high risk of extinction due to habitat loss and exploitation by the local people for its fuel and fodder values. Therefore, the present study was conducted to standardize a micropropagation protocol for Sophora mollis by using shoot tip explants and to study the meiotic chromosome count in the species. Results Multiple shoots were induced in shoot tip explants of Sophora mollis in Murashige and Skoog medium supplemented with different concentrations of cytokinins alone (BAP, TDZ, and Kinetin) and in combination with varying concentrations of NAA. MS medium supplemented with BAP (8.9 μM) was observed to be the optimal medium for multiple shoot induction and maximum 25.32 shoots per explant was obtained with average length of 4.5 ± 0.8 cm. In vitro developed shoots were transferred onto rooting media supplemented with different concentrations of auxin (IAA, IBA, and NAA). Maximum 86% rooting was observed in half-strength MS medium supplemented with 21.20 μM NAA with an average of 21.26 roots per culture. In vitro raised plantlets were adapted to greenhouse for better acclimatization and 60% plants were successfully transferred to the open environment. Based on the chromosome counts available from the literature and the current study, the species tend to show a basic chromosome number of x = 9. Conclusion The micropropagation protocol standardized can be helpful for the ex situ mass multiplication and germplasm conservation of the endangered species. Moreover, the ex situ conservation approach will be helpful in actively bridging the gap between ex situ and in situ approaches through the reintroduction of species in the wild. The cytological studies revealed the basic chromosome number x = 9 of the species.

scholarly journals An Efficient Protocol for High-frequency Direct Multiple Shoot Regeneration from Internodes of Peppermint (Mentha x piperita)

2010 ◽  
Vol 5 (12) ◽  
pp. 1934578X1000501
Author(s):  
Sanjog T. Thul ◽  
Arun K. Kukreja
Keyword(s):  
Shoot Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Ms Medium ◽  
Mentha X Piperita ◽  
Half Strength ◽  
Multiple Shoot Regeneration

A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.

scholarly journals In vitro Flowering of Shoots Regenerated from Cultured Nodal Explants

2011 ◽  
Vol 39 (1) ◽  
pp. 84 ◽  
Author(s):  
Kantamaht KANCHANAPOOM ◽  
Suttinee JINGJIT ◽  
Kamnoon KANCHANAPOOM
Keyword(s):  
Multiple Shoots ◽  
Shoot Formation ◽  
In Vitro Flowering ◽  
Nodal Explants ◽  
Callus Growth ◽  
Ms Medium ◽  
Old Field ◽  
Gypsophila Paniculata ◽  
Initial Culture

A protocol for the regeneration of Gypsophila paniculata L. using nodal explants from 2-month-old field grown plants was established. The induction of multiple shoots was best obtained on Murashige and Skoog (MS) medium supplemented with 13.3 μM BA. Callus growth was observed on MS medium containing 44.3 μM BA. Calluses were transferred to MS medium supplemented with 2, 4-D (4.5, 13.5, 22.6 μM), NAA (5.3, 16.1, 26.8 μM) or BA (4.4, 13.3, 22.1 μM) for 2 months to induce shoot formation. After 6 weeks of initial culture, multiple shoots were regenerated from calluses cultured on MS medium supplemented with 13.3 μM BA. All regenerated shoots produced roots on 16.1 μM NAA containing MS medium within 4 weeks. Rooted plantlets were hardened and established in pots at 100% survival. For induction of in vitro flowering, regenerated shoots could be induced to flower efficiently when cultured on MS medium containing 13.3 μM BA and 50 g/l sucrose.

scholarly journals In vitro Shoot Cultures of Tupistra albiflora K. Larsen

2018 ◽  
Vol 17 (5) ◽  
pp. 405-411
Author(s):  
Jiraporn PALEE
Keyword(s):  
Agar Medium ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Shoot Cultures ◽  
Ms Medium ◽  
In Vitro Shoots

To evaluate an efficient protocol for the micropropagation of Tupistra albiflora K. Larsen, the effects of N6-benzylaminopurine (BA) and naphthalene acetic acid (NAA) concentrations on multiple shoot and root induction were examined. In vitro shoots were used as the explant materials which were cultured on Murashige and Skoog (MS) agar medium supplemented with 0, 1, 2, 3 and 4 mg/L BA for 4 weeks to induce multiple shoots. It was found that the MS medium containing 3 mg/L BA induced 100 % shoot formation with the highest number of 3.2 shoots per explant (2.4-fold significantly higher than the control). For root induction, in vitro shoots were cultured on MS agar medium supplemented with 0, 1, 2, 3 and 4 mg/L NAA for 8 weeks. The results showed that the MS medium containing 1 mg/L NAA induced 100 % root formation with the highest number of 6.6 roots per explant (1.8-fold significantly higher than the control).

scholarly journals In Vitro Micropropagation of Orchid (Vanda tessellata L.) from Shoot Tip Explant

1970 ◽  
Vol 17 ◽  
pp. 139-144 ◽  
Author(s):  
MS Rahman ◽  
MF Hasan ◽  
R Das ◽  
MS Hossain ◽  
M Rahman
Keyword(s):  
Multiple Shoots ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Shoot Tip ◽  
Root Induction ◽  
Ms Medium ◽  
Culture Techniques

Context: Orchid produces a huge number of minute seeds but the seeds can not germinate easily in nature due to the lack of endosperm in the seeds is an incompatibility barrier that limits its propagation in nature. Objectives: To develop in vitro culture techniques for quick propagation of Vanda tessellate, a commercially important orchid species. Materials and Methods: Shoot tips were used as experimental materials. The explants were surface sterilized and the shoot tips were excised. The isolated shoot tips were cultured in MS medium supplemented with different concentration and combinations of auxin and cytokinin. Results: The combination of 1.5 mgl-1 NAA and 1.0 mgl-1 BAP was proved to be the best medium formulation for multiple shoot formation as well as maximum shoot elongation. The single shoots were isolated from the multiple shoots and subcultured in MS medium having NAA and IBA individually and in combinations for root induction. Maximum root induction was obtained in MS agarified medium having 0.5 mgl-1NAA and 1.0 mgl-1IBA. The well rooted plantlets were hardened successfully in the potting mixture containing coconut husk, perlite, charcoal, brick pieces in the ratio of 2:1:1:1 and eventually established under natural condition.Conclusion: An efficient regeneration protocol for micropropagation in V. tessellata through shoot tip culture has been established.Key words: Shoot tip; micropropagation; orchid.DOI: 10.3329/jbs.v17i0.7122J. bio-sci. 17: 139-144, 2009

scholarly journals In vitro propagation of muskmelon (Cucumis melo L.) from nodal segments, shoot tips and cotyledonary nodes

2015 ◽  
Vol 41 ◽  
pp. 71-77
Author(s):  
S. Parvin ◽  
M. Kausar ◽  
M. Enamul Haque ◽  
M. Khalekuzzaman ◽  
B. Sikdar ◽  
...  
Keyword(s):  
Cucumis Melo ◽  
In Vitro Propagation ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Nodal Segments ◽  
Ms Medium ◽  
Cotyledonary Nodes ◽  
Cucumis Melo L

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

scholarly journals An Efficient Method for In Vitro Shoot-Tip Culture and Sporophyte Production Using Selaginella martensii Spring Sporophyte

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 235 ◽  
Author(s):  
Kyungtae Park ◽  
Bo Kook Jang ◽  
Ha Min Lee ◽  
Ju Sung Cho ◽  
Cheol Hee Lee
Keyword(s):  
Nitrogen Sources ◽  
Culture Conditions ◽  
Shoot Tips ◽  
Shoot Tip ◽  
Soil Conditions ◽  
Ms Medium ◽  
Ex Vitro ◽  
Shoot Tip Culture ◽  
Propagation Methods

Selaginella martensii, an evergreen perennial fern that is native to South America and New Zealand, is named “frosty fern” because of its beautiful white-colored leaves and it is used as an ornamental plant. Efficient propagation methods for this species have not been developed. We aimed to develop an efficient propagation method for S. martensii through in vitro culture. We investigated culture conditions that are suitable for shoot-tip proliferation and growth. The optimum shoot-tip culture conditions were determined while using Murashige and Skoog (MS) medium (quarter, half, full, or double strength) and macronutrients (sucrose and two nitrogen sources) at various concentrations. In MS medium, the shoot tips formed a maximum of 6.77 nodes per explant, and each node formed two new shoot tips (i.e., 26 or 64 shoot tips). When using branching segments containing an angle meristem, the shoot-to-rhizophore formation ratio could be controlled by medium supplementation with plant-growth regulators. Sporophytes that were grown from shoot tips in vitro were acclimated in ex vitro soil conditions and successfully survived in the greenhouse. Numerous shoot tips could be obtained from in vitro-grown sporophytes and be proliferated ex vitro to produce a large number of plants. This method provides a way of shortening the time that is required for producing a large stock of S. martensii planting material.

scholarly journals An efficient in vitro regeneration protocol to generate stable transgenic lines of Black gram (Vigna mungo)

2018 ◽  
Vol 78 (3) ◽  
Author(s):  
Bidyut Kumar Sarmah ◽  
Trishna Konwar ◽  
Borsha Borah ◽  
Arun Kumar Handique ◽  
Sumita Acharjee
Keyword(s):  
Transformation Efficiency ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Vigna Mungo ◽  
Ms Medium ◽  
Black Gram ◽  
Transgenic Lines ◽  
Half Strength ◽  
The Individual

An efficient and quick in vitro regeneration protocol was developed for black gram (Vigna mungo) using wounded embryonic axis with cotyledon as explant. Murashige and Skoog (MS) medium supplemented with 4.44 μM BAP and 2.32 μM Kinetin was found to be effective in producing maximum number (mean 7.80) of multiple shoots. The individual shoots elongated to 4.5 cm when MS medium was supplemented with 2.89 μM GA3 along with 0.44 μM BAP and 0.46 μM KIN. A novel in vitro rooting technique was also optimized for black gram using half-strength liquid MS medium supplemented with 1.34 μM NAA. The shoots in this medium produced the highest number (mean 7.50) of roots with root length of 6.02 cm. The plantlets were transferred to soil mixture and placed in greenhouse where more than 80% successfully grew to maturity. The same protocol was successfully used to generate transgenic black gram lines carrying Bt-Cry2Aa gene through Agrobacteriummediated transformation with a transformation efficiency of 0.42%. The rooted T0 plants grew to maturity and produced T1 seeds with the presence and expression of transgene in T1 plants. Thus, we have standardized an in vitro regeneration protocol suitable for generation of stable transgenic black gram plants.

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