Development of an in vitro propagation protocol for ex situ conservation of two critically endangered species of Commersonia (Malvaceae) from Western Australia

2010 ◽  
Vol 58 (7) ◽  
pp. 565 ◽  
Author(s):  
Shahab Nikabadi ◽  
Eric Bunn ◽  
Shane Turner ◽  
Jason Stevens ◽  
Kingsley Dixon
Keyword(s):  
Endangered Species ◽  
Incubation Period ◽  
Western Australia ◽  
In Vitro Propagation ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Critically Endangered ◽  
Ms Medium ◽  
Critically Endangered Species

Protocols for in vitro propagation of two critically endangered species, Commersonia adenothalia C.F.Wilkins ms and Commersonia sp. Mt Groper (R. Cranfield & D. Kabay 9157), from south-western Western Australia were established utilising both shoot and in vitro leaf explants. Regeneration from leaf explants was highest, with an average of four shoots per leaf explant per a 4-week incubation period on ½-strength Murashige and Skoog (MS) medium with 2.5 µM thidiazuron (TDZ) + 2.5 µM 3-indoleacetic acid (IAA) for C. adenothalia and 13 shoots per leaf explant on ½-strength MS medium + 4.5 µM 6-benzylaminopurine (BAP) and 2.5 µM 1-naphthaleneacetic acid (NAA) for C. sp. Mt Groper. Shoot proliferation using single shoot explants of C. adenothalia resulted in a maximum average of 3.5 shoots per shoot explant per a 5-week incubation period on ½-strength MS medium + 5 µM kinetin and 0.5 µM BAP, whereas maximum mean shoot multiplication with C. sp. Mt Groper (×30 shoots per shoot explant per a 5-week incubation period) was recorded with ½-strength MS medium + 2.5 µM kinetin and 1 µM BAP. In general, C. sp. Mt Groper was much more reactive to cytokinins than was C. adenothalia, with prolific regeneration of shoots from leaf explants or shoot explants. Both species produced roots readily on ½-strength MS medium without added hormones or with 5 µM indole-3-butyric acid (IBA) (100% rooting in 3–4 weeks) and rooted plantlets survived the transition to soil (~70% survival).

scholarly journals In Vitro Propagation of Digitalis trojana Ivanina., an Endemic Medicinal Plant of Turkey

2020 ◽  
Author(s):  
Nurşen Çördük ◽  
Cüneyt Aki
Keyword(s):  
Acetic Acid ◽  
Medicinal Plant ◽  
In Vitro Propagation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Northwestern Turkey ◽  
Helen Of Troy ◽  
Endemic Medicinal Plant

Digitalis trojana Ivanina is a member of the Plantaginaceae family and known by its common name, Helen of Troy foxglove. It is perennial endemic to Çanakkale and Balıkesir, northwestern Turkey. In order to develop an efficient shoot regeneration protocol, the leaf explants of D. trojana were cultured on Murashige and Skoog (MS) medium containing 6-benzyl adenine (0.1, 0.5, 1.0, 3.0, 5.0 mg/L) and α-naphthalene acetic acid (0.1, 0.5, 1.0 mg/L), 3% (w/v) sucrose and 0.8% (w/v) agar. The highest number of regenerated shoots was obtained from leaf explants that were cultured on MS medium with 3.0 mg/L BA+0.1 mg/L NAA. Regenerated shoots were rooted on MS medium without plant growth regulators. Rooted plants (2–3 cm) were separately transferred to pots containing a mixture of peat and perlite (2:1 v/v) and acclimatized successfully in a growth chamber.

scholarly journals In vitro callus induction of gerbera from leaf explant

2020 ◽  
Vol 8 (1) ◽  
pp. 1
Author(s):  
Sadia Afrin Jui ◽  
Md. Mijanur Rahman Rajib ◽  
M. Mofazzal Hossain ◽  
Sharmila Rani Mallik ◽  
Iffat Jahan Nur ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Growth Regulators ◽  
Callus Induction ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Indole 3 Acetic Acid

The experiment was designed to evaluate the effect of growth regulators on leaf explant of Gerbera for callus induction. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), α-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Indole-3-acetic acid (IAA) were used to initiate cultures. These were added to Murashige and Skoog medium in different combinations and concentrations. Leaf explants cultured on MS medium supplemented with BAP+ 2, 4-D+ IAA in T4 treatment & BAP+ 2,4-D in T5 treatment showed the best results for callus induction. On the other hand callus was induced early in the combination of BA+ 2,4-D + IAA hormone in T5, T9 & T8 treatment respectively. The rate of callus induction was very low in BA + NAA combinations but it was much earlier.   

scholarly journals Micropropagation of Liriope muscari via leaf explant

1969 ◽  
Vol 83 (3-4) ◽  
pp. 169-173
Author(s):  
Keithley L. Amory ◽  
John M. Gill
Keyword(s):  
Acetic Acid ◽  
Mass Production ◽  
In Vitro Propagation ◽  
Solid Medium ◽  
Leaf Explants ◽  
Ms Medium ◽  
Isopentenyl Adenine ◽  
Young Leaves ◽  
Dichlorophenoxy Acetic Acid

Young leaves of Liriope muscari provide an ample source of explants for in vitro propagation in tropical countries where flowering is scarce. Leaves were induced to form calli on a solid medium containing Murashige and Skoog (MS) salts and vitamins, 3% sucrose, 0.7% agar, 1 mg/L 2,4-dichlorophenoxy- acetic acid (2, 4-D) and 1 mg/L 6-furfurylaminopurine (kinetin). Only the proximal segments of the leaves produced calli. These calli were induced to produce multiple plantlets on MS medium, 3% sucrose, 0.7% agar, and 10 mg/L N6 (2-isopentenyl) adenine (2 ip). It is possible to use leaf explants for in vitro mass production of Liriope. However, in variegated varieties, only green or white plants were produced, because of a chimera in the original tissue.

In Vitro Shoot Regeneration of NAA-Pulse Treated Plumular Leaf Explants of Cowpea

2010 ◽  
Vol 2 (2) ◽  
pp. 60-63 ◽  
Author(s):  
Muhammad AASIM
Keyword(s):  
Somatic Embryogenesis ◽  
Shoot Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Grain Legume ◽  
Ms Medium ◽  
Pulse Treatment ◽  
Mature Embryos ◽  
Legume Crop

Cowpea (Vigna unguiculata L.) is an economically important grain legume crop and is an important source of dietary protein in many of the developing countries. The present study reports the effect of pulse treatment duration, concentration of NAA and presence of NAA in the culture medium on shoot regeneration from plumular leaf explant of Turkish cowpea cv. ‘Akkiz’ and ‘Karagoz’. Pulse treatment of mature embryos with 20 mg l-1 NAA for 1 and 3 weeks followed by culturing of plumular leaf explant on MS medium containing 0.25, 0.50 and 1.0 BAP with 1.0, 2.0 and 4.0 mg l-1 NAA promoted somatic embryogenesis in both cultivars. Longer duration of pulse treatment was deleterious resulting in browning and consequently death of the embryos on explants. Pulse treatment with 20 mg l-1 NAA for one week was less deleterious and developed two plantlets after the explants were transferred to MS0 medium after 6 weeks through somatic embryogenesis in cv. ‘Akkiz’. Pulse treatment with 10 mg l-1 NAA for 1 week showed 33.33-50.00% and 25.00-50.00% shoot regeneration frequency in cv. ‘Akkiz’ and ‘Karagoz’ respectively on MS medium containing 0.25-1.00 mg l-1 BAP. Maximum number of 2.50 shoots each per explant were recorded in cv. ‘Akkiz’ and ‘Karagoz’ on MS medium containing 1.00 and 0.50 mg l-1 BAP respectively. Contrarily, maximum shoot length of 8.98 cm of cv. ‘Akkiz’ and 9.42 cm of cv. ‘Karagoz’ was recorded on MS medium containing 0.50 mg l-1 BAP and 1.00 mg l-1 BAP respectively. Regenerated shoots were rooted on MS medium containing 0.5 mg l-1 IBA and and acclimatized in growth room at room temperature where they produced viable seeds.

In vitro propagation of Eucalyptus phylacis L. Johnson and K. Hill., A critically endangered relict from Western Australia

2005 ◽  
Vol 41 (6) ◽  
pp. 812-815 ◽  
Author(s):  
Eric Bunn ◽  
Tissa Senaratna ◽  
Krishnapillai Sivasithamparam ◽  
Kingsley W. Dixon
Keyword(s):  
Western Australia ◽  
In Vitro Propagation ◽  
Critically Endangered

scholarly journals Efficient Regeneration of Tobacco (Nicotiana tabacum L.) Plantlets from Cotyledon, Hypocotyl and Leaf Explants: An Excellent Model Plant for Gene Function Analysis

2020 ◽  
pp. 1-9
Author(s):  
Md. Shoyeb ◽  
Kanis Fatema ◽  
Md. Abdur Rauf Sarkar ◽  
Atikur Rahman ◽  
Shaikh Mizanur Rahman
Keyword(s):  
Gene Function ◽  
Callus Induction ◽  
Function Analysis ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Regeneration Ability ◽  
Ms Medium ◽  
Gene Function Analysis

Tobacco has been widely used as a model plant for stable and non-stable gene function analysis. Successful Agrobacterium-mediated transformation mainly depends on in vitro regeneration of tobacco plant. However, a reliable and standard regeneration protocol of tobacco using multiple explants is limited. In this study, we established a reliable and reproducible regeneration protocol of tobacco using three different explants i.e. cotyledon, hypocotyl and leaf. Preliminary, surface sterilized tobacco seeds were germinated on growth regulator free MS medium. Thereafter, in vitro germinated explants were inoculated into Murashige and Skoog [1] media supplemented with different combination and types of growth regulators for callus induction and subsequent regeneration of plantlets. It was revealed that, regeneration ability of explants is greatly influenced by type and nature of the explant. Among the three explants, higher callus induction (95%) was obtained in MS medium supplemented with 2.0 mg l-1 kinetin + 2.0 mg l-1 IAA from leaf explant. Also, leaf explant exhibited much higher regeneration ability (95%) than hypocotyl (60%) and cotyledon (45%) explants. Significantly highest number of shoots (8.0) were regenerated from leaf explants cultured on MS medium supplemented with 3.0 mg l-1 Kinetin+1.0 mg l-1 IAA compared to the other hormone combinations. Regenerated mature shoots were showed normal root after transferred onto ½ MS medium containing 0.3 mg l-1 IBA. This study will provide valuable information related to in vitro regeneration of tobacco plantlets using cotyledon, hypocotyl and leaf explants and will be used as a standard protocol for Agrobacterium-mediated transformation for gene function analysis.

scholarly journals Plant Regeneration from Leaf Explants of the Medicinal Herb Wedelia chinensis

Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To
Keyword(s):  
Plant Regeneration ◽  
In Vitro Propagation ◽  
Liver Toxicity ◽  
Leaf Explants ◽  
Basal Medium ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Average Percentage

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.

scholarly journals Explants selection for in vitro propagation of Pachyrhizus erosus L.

2021 ◽  
Vol 13 (1) ◽  
pp. 10844
Author(s):  
Idowu A. OBISESAN ◽  
Ayobola M. A. SAKPERE ◽  
Bamidele J. AMUJOYEGBE ◽  
Michael S. AKINROPO
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Propagation ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stem Explants ◽  
Ms Medium ◽  
Friable Callus ◽  
Pachyrhizus Erosus ◽  
Regeneration Response

Pachyrhizus erosus tuber is rich in protein asides its agronomical value as a legume, but the seeds by which it is propagated have very low viability. This study established sterilization protocol and effect of various concentrations of auxins and cytokinins on callus production and shoot regeneration from explants of P. erosus. Explants and seeds were sterilized using sodiumhypochlorite (NaClO) solution (5, 10 and 15% v/v) for 5 and 10 mins. Nodal, stem and leaf explants from in vitro germinated P. erosus and tuber from field grown plant were sterilized and cultured on Murashige and Skoog (MS) medium (control) and MS combined with different concentrations of auxins (NAA and 2, 4-D) and cytokinin (BA and Kinetin) and the cultured explants were monitored in terms of degree of callus formation, morphology and colour of callus and also for shoot induction. The results showed that seeds of P. erosus sterilized with 10% NaClO solution for 10 mins and germinated in vitro is the best way of getting sterile nodal, stem and leaf explants for the in vitro propagation of the plant, while tuber explants could be sterilized with 15% NaClO for 10 minutes. Nodal explants inoculated in MS medium supplemented with 1.0 mg/L BA gave the highest shoot regeneration response, while stem explants inoculated on MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L NAA also gave the highest amount of friable callus. The study concluded that in vitro germinated seeds were the best way of getting explant for P. erosus.

scholarly journals Development of an In Vitro Propagation Protocol and a Sequence Characterized Amplified Region (SCAR) Marker of Viola serpens Wall. ex Ging

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 246
Author(s):  
Shipra Rani Jha ◽  
Ruphi Naz ◽  
Ambreen Asif ◽  
Mohammad K. Okla ◽  
Walid Soufan ◽  
...  
Keyword(s):  
Acetic Acid ◽  
In Vitro Propagation ◽  
Dna Analysis ◽  
Scar Marker ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Adventitious Shoot Formation ◽  
Sequence Characterized Amplified Region

An efficient protocol of plant regeneration through indirect organogenesis in Viola serpens was developed in the present study. Culture of leaf explants on MS (Murashige and Skoog) medium supplemented with 2.0 mg/L 6-benzyladenine and 0.13 mg/L 2,4-dichloro phenoxy acetic acid. Adventitious shoot formation was observed when calli were transferred on to MS medium containing 0.5 mg/L α-naphthalene acetic acid and 2.25 mg/L kinetin, which showed the maximum 86% shoot regeneration frequency. The highest root frequency (80.92%) with the 5.6 roots per explant and 1.87 cm root length was observed on MS medium supplemented with 2 mg/L indole-3-butyric acid. The plantlets were transferred to the mixture of sand, coffee husk and soil in the ratio of 1:2:1 in a pot, and placed under 80% shade net for one month. It was then transferred to 30% shade net for another one month, prior to transplantation in the field. These plantlets successfully acclimatized under field conditions. A Sequence Characterized Amplified Region (SCAR) marker was also developed using a 1135 bp amplicon that was obtained from RAPD (Random Amplification of Polymorphic DNA) analysis of six accessions of V. serpens. Testing of several market samples of V. serpens using the SCAR marker revealed successful identification of the genuine samples of V. serpens. This study, therefore, provides a proficient in vitro propagation protocol of V. serpens using leaf explants and a SCAR marker for the authentic identification of V. serpens. This study will be helpful for conservation of authentic V. serpens.

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