Clonal propagation of Chrysanthemum morifolium ramat using various explants obtained from field grown plants

A reliable and rapid large scale micropropagation method has been established from the node, shoots tip and leaf explant of Chrysanthemum morifolium growing in field condition. Experiments were conducted to standardize the culture media with plant hormone for multiple shoot proliferation and rooting for obtaining plantlets with uniform characteristics like mother plant in terms of growth and habits. Different concentrations and combinations of auxins (IAA) and cytokinins (BAP, Kin) were used in MS for the above purpose. Maximum shoot regeneration was found in MS treated with 2.0 mg/l BAP both in node and shoot tip explants. In the above combination, nodal explants produced 16 initial shoots. Shoot tip explants produced 12 shoots and leaf segment produced 07 shoots. For in vitro rooting, different concentrations of IBA and NAA were used. Higher rooting percentage was recorded on MS fortified with 1.5 mg/l IBA. The rooted plantlets were hardened and successfully established in the soil. About 90% of the regenerated plantlets survived in the natural environment.

Download Full-text

In vitro shoot proliferation of Passiflora caerulea L. via cotyledonary node and shoot tip explants

BioTechnologia ◽

10.5114/bta.2017.68310 ◽

2017 ◽

Vol 2 ◽

pp. 113-119 ◽

Cited By ~ 1

Author(s):

Marziyeh Jafari ◽

Mohammad Hosein Daneshvar ◽

Amin Lotfi

Keyword(s):

Shoot Proliferation ◽

Cotyledonary Node ◽

Shoot Tip ◽

Shoot Tip Explants ◽

In Vitro Shoot Proliferation

Download Full-text

In vitro Mass Propagation of a Ground Orchid - Phaius tancarvilleae (L'Her.) Blume through Shoot Tip Culture

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v21i2.10241 ◽

2011 ◽

Vol 21 (2) ◽

pp. 181-188 ◽

Cited By ~ 3

Author(s):

Bijaya Pant ◽

Sumitra Shrestha

Keyword(s):

High Frequency ◽

Shoot Proliferation ◽

Shoot Multiplication ◽

Multiple Shoots ◽

Shoot Tip ◽

Mass Propagation ◽

Shoot Tip Culture ◽

Shoot Tip Explants ◽

Direct Shoot

High frequency direct shoot proliferation was induced from the shoot tip explants derived from the in vitro grown seedlings of a critically endangered and horticulturally important ground orchid Phaius tancarvilleae (L'Her) Blume. Shoot tip explants cultured on solidified MS with alone or combination of various concentrations of NAA and BAP produced shoots and multiple shoots. The maximum number of healthy shoots was observed on MS with BAP (1.0 mg/l) with an average of 13.3 shoots per culture in 20 weeks; where shoot multiplication was initiated after 4 weeks of culture. Regenerated shoots rooted on MS with various concentrations of NAA, IAA, IBA. MS with NAA (0.5 mg/l) was the most appropriate condition for rooting. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing cocopeat and sphagnum moss in the ratio of 2 : 1. Key words: Mass propagation, Phaius tancarvilleae, shoot multiplication D. O. I. 10.3329/ptcb.v21i2.10241 Plant Tissue Cult. & Biotech. 21(2): 181-188, 2011 (December)

Download Full-text

In vitro Mass Propagation of Artemisia (Artemisia annua L.) cv: Anamed

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v23i2.17518 ◽

2014 ◽

Vol 23 (2) ◽

pp. 165-176 ◽

Cited By ~ 4

Author(s):

Tilahun Hailu ◽

Balcha Abera ◽

Gabra Mariam

Keyword(s):

Large Scale ◽

Artemisia Annua ◽

Shoot Tip ◽

Nodal Explants ◽

Mass Propagation ◽

Mean Values ◽

Coffee Husk ◽

Soil Mix ◽

Shoot Tip Explants

An efficient in vitro propagation protocol was developed for anamed (A-3) cultivar of Artemisia annua. Two and 1.5% concentration of NaOCl treatment for 10 and 20 min were found to be optimum for sterilization of shoot tip and nodal explants, respectively. Maximum percentage (98.75 ± 2.50) shoot induction was observed from nodal explants cultured on MS supplemented with 0.8 mg/l BAP + 0.1 mg/l IBA followed by 82.50 ± 2.88% from shoot tip explants on the same medium with 0.8 mg/l TDZ for shoot tip explants. The highest number of shoots (8.05 ± 0.66/explant) was regenerated on MS + 1 mg/l BAP + 0.1 mg/l IBA. Best rooting with mean values of 18.25 ± 0.95/explant root number and root length (6.35 ± 0.10 cm) was recorded on 1/2 MS + 0.5 mg/l IBA. Up on acclimation and transplanting, 80% survival efficiency was observed on the soil mix ratio of 2 : 1 : 1 (decomposed coffee husk, forest soil and sand, respectively). The developed regeneration protocol enables a large scale commercial production and a possible system towards the genetic improvement of this crop. D. O. I. http://dx.doi.org/10.3329/ptcb.v23i2.17518 Plant Tissue Cult. & Biotech. 23(2): 165-176, 2013 (December)

Download Full-text

PENGARUH EKSPLAN DAN ZPT TERHADAP PERTUMBUHAN nEPEnTHES ALBOmARGInATA SECARA In VITRO

Jurnal Teknologi Lingkungan ◽

10.29122/jtl.v12i1.1267 ◽

2016 ◽

Vol 12 (1) ◽

pp. 103

Author(s):

Lazarus Agus Sukamto

Keyword(s):

Multiple Shoots ◽

Carnivorous Plant ◽

Multiple Shoot ◽

Shoot Tip ◽

Naphthalene Acetic Acid ◽

Dichlorophenoxyacetic Acid ◽

Over Exploitation ◽

2,4 Dichlorophenoxyacetic Acid ◽

Shoot Tip Explants

Nepenthes albomarginata Lobb ex Lindl. is a carnivorous plant, distributes in several regions in Indonesia. The plant population decreases drastically because of over exploitation and ruining nature habitat. Plant propagation by nature and cutting are not enough to rehabilitation its population. In vitro culture of N. albomarginata was carried out using plantlets grown from the seeds in vitro. Plantlets were cut to became two part explants, consisted of shoot tip and under-shoot tip cuttings. These cutting explants were grown on Murashige & Skoog (MS) media with addition of plant growth regulators of 6-benzylaminopurine (BA), combined with or without-naphthalene acetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D) at 1 mg/l. Shoot tip cuttings of N. albomarginata formed double multiple shoot 25,00% on control; formed triple multiple shoots 25,00% onBA 1 mg/l treatment; formed callus 37,50%, triple or quartet shoots 25,00% and rooted plantlets 25,00% on BA 1 mg/l + NAA 1 mg/l treatment. The under-shoot tip cuttings ofN. albomarginata formed double – triple shoots 25,00% and rooted plantlets 37,50% on control; formed double – triple shoots 25,00% and rooted plantlets 12,50% on BA 1 mg/ltreatment; formed callus 12,50%, double - pentacle shoots 37,50% and rooted plantlets 25,00% on BA 1 mg/l + NAA 1 mg/l treatment. 2,4-D 1 mg/l or its combined with BA 1mg/l treatment caused deadly shoot tip or under-shoot tip explants. The combination of BA 1 mg/l + NAA 1 mg/l was the best treatment for producing callus, multiple shootsand rooted plantlets of N. albomarginata.

Download Full-text

In vitro plant regeneration of Paulownia tomentosa (Thunb.) Steud. from shoot tip and leaf segment

Bangladesh Journal of Botany ◽

10.3329/bjb.v44i3.38555 ◽

2018 ◽

Vol 44 (3) ◽

pp. 459-463

Author(s):

PK Roy

Keyword(s):

Multiple Shoots ◽

Shoot Elongation ◽

Multiple Shoot ◽

Shoot Tip ◽

Leaf Segment ◽

Induction Medium ◽

Paulownia Tomentosa ◽

Shoot Induction Medium ◽

Number Of Shoots

An efficient protocol was developed for in vitro mass propagation of Paulownia tomentosa (Thunb.) Steud. using shoot tip and leaf segment explants from field grown plant. Different concentrations and combinations of BAP, Kn, zeatin and NAA were used for multiple shoot regeneration. Among two types of explants, leaf segment produced the highest number of shoots per explant (12 ± 0.4) when they were cultured on MS supplemented with 3.0 mg/l Kn and 0.5 mg/l NAA. Addition of 10% CW to above mentioned medium increased the number of shoots (18) per culture. Shoot tip explants also produced multiple shoots in the same medium, but their performance was not good as leaf segment explants. For shoot elongation, 100 mg/l urea was more effective when added with best shoot induction medium. Shoots rooted well in halfstrength MS supplemented with 2.0 mg/l NAA, within 12 - 15 days. Regenerated plantlets were successfully acclimatized and established in poly bag containing a mixture of soil and compost in 2:1 ratio. About 90% plantlets survived under open field conditions.

Download Full-text

Assessing the Efficacy of Broad-Spectrum Antibiotics in Controlling Bacterial Contamination in the In Vitro Micropropagation of Ginger (Zingiber officinale Rosc)

The Scientific World JOURNAL ◽

10.1155/2020/6431301 ◽

2020 ◽

Vol 2020 ◽

pp. 1-8

Author(s):

Selam Tewelde ◽

Subban Patharajan ◽

Zenebe Teka ◽

Desta Berhe Sbhatu

Keyword(s):

Broad Spectrum ◽

Bacterial Contamination ◽

Culture Media ◽

Zingiber Officinale ◽

Shoot Tip ◽

Broad Spectrum Antibiotics ◽

Bacterial Contaminants ◽

Planting Materials ◽

Shoot Tip Explants

Ginger (Zingiber officinale Rosc) (Zingiberaceae) is a livelihood and commercial crop in Ethiopia. But, the availability of clean and healthy planting materials has become a problem due to wilt disease, caused by Ralstonia solanacearum Biovar 3 Race 4. This problem obliged growers to seek for tens of millions of vigorous and disease-free planting materials very quickly via in vitro micropropagation of shoot tip explants. For this purpose, protocols of sterilizing shoot tip explants and controlling bacterial contamination of one Ethiopian ginger cultivar called Deribo were tested. Hence, this article reports the finding of a study that aimed at testing the (a) effectiveness of three sterilization agents, namely, 0.25% w/v RBK (composed of ridomile, bayleton, and kocide at 1 : 1 : 1 ratio), 0.50% v/v NaOCl, and 70% v/v ethanol at three different treatment times in combination with 0.25% HgCl2; (b) efficacy of four broad-spectrum antibiotics and their combinations in controlling bacterial contaminants of ginger shoot tip explants and in vitro micropropagation media; and (c) effects of the antibiotics on the shooting performances of the explants of the cultivar. A 0.50% v/v NaOCl at exposure time of 20 min followed by 0.25% HgCl2 has resulted in 80% contamination-free and 70% live explants after three weeks of incubation. Likewise, cefotaxime at 50, 100, and 200 mg/L and cefotaxime plus streptomycin at 25, 50, and 100 mg/L yielded 87 to 93% contamination-free microshoots after three weeks of culturing. The number of explants killed by the antibiotics increased with increasing the concentration of the antibiotics. Cefotaxime at 50 mg/L and cefotaxime plus streptomycin at 25 mg/L yielded significantly highest mean microshoots per explant (7.10 ± 0.36 and 7.51 ± 0.27, respectively) and mean shoot length (4.2 ± 0.26 and 3.56 ± 0.17 cm, respectively). Some of the microshoots showed some yellowing. But, they turned green and grew normal after subcultured into fresh, antibiotics-free culture media. These findings are important foundations towards developing more optimized protocols of sterilizing explants and controlling bacterial contaminants for large-scale in vitro micropropagation of the Deribo ginger cultivar.

Download Full-text

Thidiazuron and Benzylamino Purine Stimulation of Apple Shoot Proliferation in Vitro

HortScience ◽

10.21273/hortsci.31.4.629e ◽

1996 ◽

Vol 31 (4) ◽

pp. 629e-629

Author(s):

Karim H. Al-Juboory ◽

Jabar H. Al-Niami

Keyword(s):

Malus Domestica ◽

Shoot Proliferation ◽

Shoot Tip ◽

Ms Medium ◽

Proliferation In Vitro ◽

Shoot Tip Explants ◽

Apple Shoot ◽

Stimulation Of

Thidiazuron (TDZ) and benzylamino purine stimulated shoot proliferation on shoot tip explants of wild apple (Malus domestica Borkh) when incorporated in Murashige and Skoog (MS) medium at concentrations of 1.0–10 μm. Shoot numbers obtained with TDZ were greater than the number produced when using BA in the medium but the shoots were shorter than with BA. Increasing TDZ levels increased shoot proliferation with 10 μm. Apple shoots were successfully rooted on MS medium with 2.0 mg·L–1 NAA and then transferred to a mixture of 1 peat: 1 perlite: 1 soil and acclimatized for potting.

Download Full-text

An improved micropropagation protocol for lentisk (Pistacia lentiscus L.)

Folia Horticulturae ◽

10.2478/fhort-2019-0003 ◽

2019 ◽

Vol 31 (1) ◽

pp. 61-69

Author(s):

Hakan Yildirim ◽

Ahmet Onay ◽

Kazim Gunduz ◽

Sezai Ercisli ◽

Firat Ege Karaat

Keyword(s):

In Vitro Propagation ◽

Large Scale ◽

Shoot Proliferation ◽

Multiple Shoot ◽

Shoot Length ◽

Pistacia Lentiscus ◽

Shoot Induction ◽

Multiple Shoot Induction ◽

Long Shoots

AbstractThis study presents an efficient improvement in the in vitro propagation protocol for one cloned genotype of lentisk (Pistacia lentiscus L.) by assessing the effects of gibberellic acid (GA3) concentrations, different cytokinins and amino acids and their concentrations on shoot proliferation, the effects of shoot length on rooting and the effects of compost type (sterile and non-sterile) on acclimatization. The best growth medium for multiple shoot induction was the MS medium supplemented with a combination of 1 mg l−1 BA, 100 mg l−1 tryptophan and 0.5 mg l-1 GA3, which gave a mean shoot length of 1.64 ± 0.07 cm and a mean bud number of 5.46 ± 0.16. The best results in terms of root length, rooting rate and the number of roots per shoot were obtained with 2 cm long shoots. The rooted plantlets were readily acclimatized in the sterile compost. In conclusion, the micropropagation protocol developed in this study can be used for large-scale propagation of P. lentiscus L. in reforestation programmes.

Download Full-text

Utilization of Aseptic Seedling Explants for In vitro Propagation of Indian Red Wood

Notulae Scientia Biologicae ◽

10.15835/nsb549188 ◽

2013 ◽

Vol 5 (4) ◽

pp. 518-523 ◽

Cited By ~ 2

Author(s):

Kishore Kumar CHIRUVELLA ◽

Arifullah MOHAMMED ◽

Rama Gopal GHANTA

Keyword(s):

Large Scale ◽

Ex Situ Conservation ◽

Cotyledonary Node ◽

Multiple Shoot ◽

Shoot Tip ◽

Ex Situ ◽

Nodal Segments ◽

Ms Medium ◽

Shoot Differentiation

Micropropagation has been advocated as one of the most viable biotechnological tool for ex situ conservation of rare, endangered endemic medicinal plants germplasm. Rapid clonal micropropagation protocol for large-scale multiplication of an endemic medicinal plant Soymida febrifuga (Meliaceae) was established from 15-day aseptic seedling cotyledonary node and shoot tip explants. High frequency of sprouting and shoot differentiation was observed from cotyledonary node explants compared to shoot tip, on Murashige and Skoog (MS) medium fortified with BA, KN, 2-iP and CM. Of the cytokinins used, BA (3.0 mgl-1) supported highest average number and maximum multiple shoot differentiation (16.6). In vitro proliferated shoots were multiplied rapidly by culturing nodal segments as microcuttings, further subcultured on the same media for elongation. Elongated shoots upon transfer to MS medium fortified with IBA showed rooting within two weeks of culture. Rooted plantlets were successfully hardened and 75% of rooted shoots successfully survived on establishment to the soil. Plants looked healthy with no visually detectable phenotypic variations. This protocol provides a successful and rapid technique that can be used for ex situ conservation minimizing the pressure on wild populations and contributes to the conservation of this endemic medicinally potent flora.

Download Full-text