scholarly journals Effect of Alkylating Mutagens on Rooting Response and Callus Age on Shoot Regeneration of Rough Lemon (Citrus Jambhiri Lush.)

2020 ◽  
pp. 1-10
Author(s):  
Mudasir Iqbal ◽  
Parshant Bakshi ◽  
B. K. Sinha ◽  
Mohsin Iqbal ◽  
Arti Devi
Keyword(s):  
Farmyard Manure ◽  
Climatic Conditions ◽  
High Yield ◽  
Garden Soil ◽  
In Vitro Mutagenesis ◽  
Ms Medium ◽  
Citrus Jambhiri ◽  
Methane Sulphonate ◽  
Citrus Rootstock

Many species of Citrus and compatible sexual relatives are being used to develop biotic and abiotic tolerant rootstocks and their ability to confer positive stionic effects. Citrus jambhiri is the commercial citrus rootstock in India, deep-rooted well adapted to the diverse agro-climatic conditions. It ensures high yield with large size fruits in most of the scion cultivars and at the same time is resistant to most of the viruses. For in vitro mutagenesis, leaf and epicotyl calli derived shoots were used as explant material. In-vitro mutagenesis is a valuable tool for improvement of a crop, especially when there is a need to add one or two easily identifiable characters in an otherwise well adapted variety, without disturbing its basic genotype. The alkylating agent methyl methane sulphonate (MMS) and ethyl methane sulphonate (EMS) at 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6%, were used for mutagenesis. The mutagenic calli derived shoots were regenerated on MS medium augmented with BAP (3.0 mg/l), followed by rooting in MS medium containing NAA (2.0 mg/l). Percent rooting (29.50-8.33%), (27.11-07.72%), number of roots per shoot (3.11-1.18), (3.12-1.04) and root length (4.13-2.22), (4.15-2.17) decreased with increasing doses of MMS and EMS treatments, respectively. Effect of increasing age of callus showed that callus retained regeneration capacity (3.55%) even after 210 days of culture by repeated sub-culturing. The plantlets were successfully acclimatized in different potting mixtures and highest survival rate (90.35%) was achieved in potting mixture containing garden soil with sand and farmyard manure (1:1:1).

scholarly journals In vitro plant regeneration in rough lemon (Citrus jambhiri Lush.) through epicotyl segments by direct shoot organogenesis

2016 ◽  
Vol 8 (2) ◽  
pp. 724-729
Author(s):  
Sukhjit Kaur
Keyword(s):  
In Vitro Regeneration ◽  
Growth Hormones ◽  
Garden Soil ◽  
Direct Regeneration ◽  
Ms Medium ◽  
Citrus Jambhiri ◽  
Rough Lemon ◽  
Induction Frequency ◽  
Bud Induction

The effect of Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of growth hormones on direct regeneration from one month old epicotyl segments of in vitro grown rough lemon (Citrus jambhiri Lush.) seedlings was studied. The earliest bud induction in 7.5 days, highest bud induction frequency (98.50%), percent regeneration(90.53) were obtained on MS medium supplemented with 6-Benzylaminopurine (BAP) (1mglit-1) with an average number of 12.50 buds per explants. The epicotyls segments with proliferated buds were transferred to elongation media in order to improve the recovery of normal shoots. Maximum number of elongated shoots (8.50) was obtained on MS medium having BAP (0.5mglit-1) + Gibberellic Acid (GA3)(1.0 mglit-1).These elongated shoots were then rooted on MS medium containing Indole-3-butyric acid (IBA) (0.1mglit-1) + Indole-3-aceticacid(IAA)(0.5mglit-1) with highest rooting percentage(96%) and root number(5.0). Early (10.10 days) rooting was observed in MS medium supplemented with NAA1.0 mglit-1 + IBA0.5 mglit-1.The plantlet survival was 98.52%, when plantlets were transferred to plastic pots containing a mixture of garden soil and vermiculite (1:1). The hardened plants were successfully established in the soil. The present study developed protocol which can be reliably used for in vitro regeneration of rough lemon and for gene transfer studies in rough lemon, especially to induce salinity and Phytophthora tolerance.

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 Impact of Ethyl Methane Sulphonate Mutagenesis in Artemisia vulgaris L. under NaCl Stress

BioTech ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 18
Author(s):  
Sudheeran Pradeep Kumar ◽  
B.D. Ranjitha Kumari
Keyword(s):  
Nacl Stress ◽  
In Vitro Mutagenesis ◽  
Artemisia Vulgaris ◽  
Salt Tolerant ◽  
Ethyl Methane Sulphonate ◽  
Ethyl Methane ◽  
Methane Sulphonate ◽  
Constant Form ◽  
Brown Discoloration

The present investigation aimed to obtain salt-tolerant Artemisia vulgaris L. to develop a constant form through in vitro mutagenesis with ethyl methane sulphonate (EMS) as the chemical mutagen. NaCl tolerance was evaluated by the ability of the callus to maintain its growth under different concentrations, ranges from (0 mM to 500 mM). However, NaCl salinity concentration at (500 mM) did not show any development of callus, slight shrinking, and brown discoloration taking place over a week. Thus, all the biochemical and antioxidant assays were limited to (0–400 mM) NaCl. On the other hand, selected calluses were treated with 0.5% EMS for 30, 60, and 90 min and further subcultured on basal media fortified with different concentrations of 0–400 mM NaCl separately. Thus, the callus was treated for 60 min and was found to induce the mutation on the callus. The maximum salt-tolerant callus from 400 mM NaCl was regenerated in MS medium fortified with suitable hormones. Biochemical parameters such as chlorophyll, carotenoids, starch, amino acids, and phenol contents decreased under NaCl stress, whereas sugar and proline increased. Peroxidase (POD) and superoxide dismutase (SOD) activities peaked at 200 mM NaCl, whereas catalase (CAT) was maximum at 100 mM NaCl. Enhanced tolerance of 0.5% the EMS-treated callus, attributed to the increased biochemical and antioxidant activity over the control and NaCl stress. As a result, the mutants were more tolerant of salinity than the control plants.

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

Rapid Micropropagation Protocol of Mucuna pruriens var. utilis using Cotyledonary Node Explant: A Cultivated Medicinal Edible Legume

2021 ◽  
Author(s):  
Sanjay Kumar Madkami ◽  
Arpita Moharana ◽  
Durga Prasad Barik
Keyword(s):  
Seed Germination ◽  
Large Scale ◽  
Shoot Multiplication ◽  
Cotyledonary Node ◽  
Production Method ◽  
Commercial Production ◽  
Mucuna Pruriens ◽  
Garden Soil ◽  
Ms Medium

Background: The presence of L-dopa coupled with rich protein and amino acid marked Mucuna pruriens var. utilis as an important under-utilised legume. Therefore, it is useful to develop a method for large-scale multiplication for commercial production. Method: Tissue culture technology is successfully utilized in propagation of plants with poor and uncertain response to conventional propagation. Murashige and Skoog’s (MS) medium without any Plant Growth Regulators (PGRs) was used for seed germination and with PGRs for shoot and root multiplication.Result: Highest 95% seed germination was found in fresh seeds at 7-8 days of culture. Shoot multiplication percentage was found to be 100% with highest c.a. 21.1 shoots with an average 4.8 cm shoot length on MS + BAP 1.5 mg L-1 per 10 days old cotyledonary node explant. A total c.a. 144 shoots were harvested after 3rd harvest of the mother cotyledonary node with two whole cotyledons at day 70. Rooting was best induced in in vitro derived shoots on MS medium without any PGRs and plantlets were acclimatized in sand and soil (1:1) and established in pot with garden soil. 

scholarly journals Mass propagation of Bambusa bambos (L.) Voss through in vitro culture

2017 ◽  
Vol 5 (2) ◽  
pp. 15-26 ◽  
Author(s):  
Raihan I Raju ◽  
Shyamal K Roy
Keyword(s):  
In Vitro Culture ◽  
Basal Medium ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Mass Propagation ◽  
Surface Sterilization ◽  
Half Strength

Protocol for mass propagation of Bambusa bamboos (L.) Voss was developed through in vitro culture. Nodal segments containing pre-existing axillary bud, after surface sterilization, were inoculated on liquid Murashige and Skoog’s (MS) basal medium containing different concentrations and combinations of cytokinins (BAP, TDZ and Kn). The highest direct shoot induction (90%) was obtained in the MS liquid medium supplemented with 2.0 mg/l BAP and 1.0 mg/l TDZ with maximum average number of shoots (3.14 ± 0.06) per explant. Highest shoot multiplication (16.58 ± 0.24 shoots per culture) with highest average shoot length (9.21 ± 0.13 cm) was obtained when in vitro raised shoots were cultured in gelrite gelled MS medium in conjunction with 2.0 mg/l BAP and 1.0 mg/l TDZ. Incorporation of 10% coconut water with 4% sucrose in the above mentioned medium resulted satisfactory shoot growth and development with an average 26.7 ± 0.60 shoots per culture. For root induction, in vitro raised shoots were divided into clumps of 4-5 shoots in each clump and transferred onto both liquid and gelled half-strength MS medium containing different concentrations and combinations of auxins (IBA and NAA). Maximum rooting (86.67%) was achieved in half-strength of MS medium fortified with 2.5 mg/l IBA and 2.5 mg/l NAA with an average 8.72 ± 0.42 root per shoot. The rooted plantlets were then transferred to polybags containing garden soil, sand and compost mixture with 1:1:1 ratio. After a month the hardened plantlets were then transferred to the larger pots containing garden soil and compost with 1:1 ratio for sufficient growth and finally transplanted to the field. In this process, the highest 100% survivability was recorded from well-established rooted plantlets. The regenerated plants showed well developed root and shoot systems in field condition.Jahangirnagar University J. Biol. Sci. 5(2): 15-26, 2016 (December)

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

2020 ◽  
pp. 22-29
Author(s):  
Priyanka Sharma ◽  
Bidhan Roy
Keyword(s):  
Germplasm Conservation ◽  
Synthetic Seeds ◽  
Shoot Tips ◽  
Nodal Segments ◽  
Synthetic Seed ◽  
Ms Medium ◽  
Meristematic Tissue ◽  
Citrus Jambhiri ◽  
Cacl2 Solution

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.

scholarly journals In Vitro Clonal Propagation of Nardostachys jatamansi: A Traditional Himalayan Medicinal Plant

2021 ◽  
Vol 16 (3) ◽  
Author(s):  
Hem Chandra Pant ◽  
Harsh Vardhan Pant ◽  
Arun Kumar ◽  
Himani Tomar ◽  
Manish Dev Sharma ◽  
...  
Keyword(s):  
Tap Water ◽  
Shoot Proliferation ◽  
Culture Method ◽  
Multiple Shoot ◽  
Shoot Tip ◽  
Nodal Explants ◽  
Garden Soil ◽  
Ms Medium ◽  
Bud Induction

Plant tissue culture method has an impressive technique for Investigation and Explains basic and applied problems in plant biotechnology field. Micropropagation has played a vital role in the rapid multiplication of many plants species. The nodal explants and shoot tip of N. jatamansi inoculated in MS medium (Murashige and Skoog) contain different types concentrations of PGRs (Phytohormones) at various frequencies for the optimization of growth quality for shoot bud Induction, shoot proliferation and micro rooting in plant. The perfect shoot induction takes place in the concentration of BAP + IBA (2.0 mg/l +1.5 mg/1) multiplication of nodal explants and shoot tip in the combination lower concentration of BAP and KN (2.0 mg/1+1.5 mg/1) This combination proved best for multiple shoot formation. Half strength (1/2) of the MS medium containing NAA and BAP (1.5 mg/1+1.0 mg/1) in combination was most useful for rooting in plant. Well developed rooted micro shoots were smoothly removed for the culture flask and dipped in 70% ethanol for 2 min and then washed with running tap water for 5-10 min to remove media for the root and transferred to small plastic cups carry cocopeat, garden soil and sand (2:2:1) and produce healthy growth in ex-vitro conditions.

scholarly journals Regeneration of Asparagus officinalis L. Through Embryogenic Callus

2017 ◽  
Vol 27 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Mustafa Abul Kalam Azad ◽  
Muhammad Nurul Amin
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Shoot Proliferation ◽  
Asparagus Officinalis ◽  
Garden Soil ◽  
Ms Medium ◽  
Regeneration System ◽  
Embryo Germination ◽  
Embryogenic Calli

A plant regeneration system was established from hypocotyl explants of in vitro grown seedlings of A. officinalis and in vitro proliferated shoots, respectively through somatic embryogenesis and embryogenic calli. Somatic embryogenesis was significantly influenced by the types of plant growth regulators. Embryogenic calli with somatic embryos developed well in MS supplemented with 2.0 ‐ 4.0 μM BAP and 1.0 ‐ 4.0 μM 22,4‐D, NAA or IBA. The highest frequency (95.3%) of embryogenic calli and 55.2 somatic embryos formation were obtained when the MS was amended with 4.0 μM BAP and 2.0 μM 2,4‐D. The best embryo germination occurred in 1.0 μM BAP supplemented MMS. The highest 97.2% of shoot proliferation was observed in embryogenic calli in MS medium containing 2.0 μM BAP and 1.0 μM IBA. In vitro grown shoots were rooted in MMS with 1.0 ‐ 2.0 μM IBA. Regenerants were transferred to vermicompost and successfully established under an ex vitro environment in garden soil with 80% survival rate.Plant Tissue Cult. & Biotech. 27(1): 21-31, 2017 (June)

scholarly journals ESTABLISHMENT OF IN VITRO PROPAGATION OF Hibiscus cannabinus (KENAF)

2021 ◽  
Vol 5 (1) ◽  
pp. 05-07
Author(s):  
Nor Syafawati Mohamad Pauzi ◽  
Nurul Ain Saipul Bahari ◽  
Zarina Zainuddin
Keyword(s):  
In Vitro Propagation ◽  
Climatic Conditions ◽  
Optimum Concentration ◽  
Hibiscus Cannabinus ◽  
Root Induction ◽  
Ms Medium ◽  
The Past ◽  
Wide Range ◽  
Alternative Resource

Hibiscus cannabinus or commonly known as kenaf is a versatile plant that serves as resources for numerous manufacturing and livestock industries. Originally planted in West Africa, kenaf is now distributed in many countries including Malaysia as its fibres were proved to be an ultimate alternative resource for major industries such as automotive, paper and bio-composite. In fact, in Malaysia, due to its adaptation to wide range of climatic conditions, kenaf has potentially be chosen as a new industrial crop replacing tobacco. There have been many interests on regenerating kenaf via micropropagation as the demand for this crop has been increasing tremendously since the past decades. Hence, this study is initiated with the objective to establish in vitro propagation system of H. cannabinus. The callus induction was achieved on Murashige and Skoog (MS) media supplemented with different concentrations of benzylaminopurine (BAP). It was observed that calli were successfully induced on all the BAP concentrations tested. The optimum concentration of BAP that induced the healthiest and biggest calli was 3.0 mg/l. Shoot and root induction from the calli were attempted using MS medium supplemented with different combinations and concentrations of IBA, BA and GA3. From the seven treatments, three treatments successfully induced formation of shoot; treatment T3 (MS + 1.0 mg/l IBA + 2.5 mg/l BA), treatment T5 (MS + 0.1 mg/l IBA + 2.0 mg/l BA + 0.3 mg/l GA3) and treatment T6 (MS + 1.0 mg/l IBA + 2.5 mg/l BA + 0.3 mg/l GA3). The results obtained in this study can paved for more research on tissue culture of H. cannabinus.

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