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 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 The effects of genotypes and media composition on callogenesis, regeneration and cell suspension culture of chamomile (Matricaria chamomilla L.)

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11464
Author(s):  
Aqeel Ahmad ◽  
Muhammad Tahir ul Qamar ◽  
Almeera Shoukat ◽  
Mehtab Muhammad Aslam ◽  
Mohsin Tariq ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
In Vitro Regeneration ◽  
Sustainable Production ◽  
Conservation Strategy ◽  
Direct Regeneration ◽  
Ms Medium ◽  
Indirect Regeneration

Background Chamomile is an important herb being used widely for medicinal purposes. Its multitherapeutic, cosmetic, and nutritional values have been established through years of traditional and scientific use and research. Increased use of medicinal plants necessitates rational use as well as sustainable production of such genetic resources. Plant in vitro micro-propagation poses unique opportunities for sustainable production of medicinal herbs, their regrowth and conservation. The present study aimed to investigate the effects of different explants, plant growth regulators (PGRs) combinations and media type on callogenesis, in vitro regeneration and cell suspension of six chamomile genotypes to enhance its sustainable production. Methods The shoot, lateral sprout, and leaf derived explants of six chamomile genotypes including Isfahan, Shiraz, Kazeron, Goral, Sharokashari and Presso were used for direct and indirect regeneration. For indirect regeneration various doses of NAA and kinetin were used to induce calli which were cultured on MS media containing PGRs for direct and indirect regeneration. Later, cell suspension was established and morphological characterization of CrO3 stained cells was carried out using microscopy. Results and Discussion Our findings revealed that the highest callus percentage and callus volume were observed from lateral sprouts and shoots of genotype Isfahan on MS medium containing 1 mg/L NAA and 1 mg/L kinetin. The in vitro regeneration was found to be genotype dependent while 77% and 77.5% was the highest percentage for indirect and direct regeneration, respectively. Additionally, the maximum shoot number (two shoots/explant) and shoot length (2.22 cm) were also observed in Isfahan genotype. Cell suspension culture showed the highest fresh weight (18.59 g) and dry weight (1.707 g) with 0.75 g inoculum of the callus derived from lateral sprouts cultured on MS medium. Microscopy of CrO3 stained cells was carried on each 3rd day for 27 days that revealed larger and spongier cells in the early days as compared to final days when the cell number was greater but cell size was smaller. Conclusion The callogenesis, organogenesis, and cell suspension culture of chamomile may be genotype dependent. Hence, optimization of media ingredients and culture conditions is of utmost importance for devising tissue culture based conservation strategy of any chamomile genotype and secondary metabolite production.

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 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 regeneration of an important medicinal plant Centella asiatica L. Urban

2012 ◽  
Vol 47 (3) ◽  
pp. 269-272 ◽  
Author(s):  
Dr Jaheduzzaman ◽  
MA Habib ◽  
S Akter ◽  
NA Banu ◽  
RB Rahman ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Centella Asiatica ◽  
Multiple Shoot ◽  
Garden Soil ◽  
Nodal Segment ◽  
Root Induction ◽  
Ms Medium ◽  
Multiple Shoot Regeneration

An in vitro multiple shoot regeneration protocol has been developed for Centella asiatica L. Urban from the nodal and shoot tips explants using MS medium with different concentrations and combinations of growth regulators. Multiple shoot regeneration was observed from both the explants on MS containing either BAP alone or in combination with IAA, NAA and Kn. Of the two explants, nodal segment showed comparatively better response towards multiple shoot regeneration. Maximum multiple shoots were found on MS supplemented with 1.0 mg/L BAP and 0.4 mg/L NAA. For root induction, well-developed shoots were excised and cultured on both MS and half strength of MS with various concentrations of IBA, IAA and NAA. Profuse healthy rooting was obtained on MS medium containing 0.2 mg/L IBA. The well rooted plantlets were successfully transplanted to the garden soil and their survival rate under natural condition was 90-95 %. DOI: http://dx.doi.org/10.3329/bjsir.v47i3.13058 Bangladesh J. Sci. Ind. Res. 47(3), 269-272, 2012

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 In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Genetic Transformation ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Native Forest ◽  
Ms Medium ◽  
Dalbergia Sissoo ◽  
Callus Induction Medium

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.

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