scholarly journals FAST DIRECT REGENERATION OF PLANTS FROM NODAL EXPLANTS OF Stevia rebaudiana Bert.

2019 ◽  
Vol 18 (5) ◽  
pp. 95-103
Author(s):  
Romuald Doliński ◽  
Krzysztof Kowalczyk
Keyword(s):  
Basal Medium ◽  
Stevia Rebaudiana ◽  
Production Costs ◽  
Nodal Explants ◽  
Direct Regeneration ◽  
Naphthaleneacetic Acid ◽  
Production Scale ◽  
Plant Weight ◽  
The Media

In the preceding research, stevia has been typically cloned in vitro using two media, on which the shoots were formed (3–6 weeks), and on the other they were rooted (3–5 weeks). This study aimed at finding the possibility for rapid stevia propagation from large nodal explants using the MS basal medium [Murashige and Skoog 1962], with low auxin concentrations (0.5, 1 and 2 mg‧dm–3). The plants were obtained as soon as after three weeks. The best results were obtained from media with various concentrations of the indole-3-acetic acid (IAA) and the highest concentration of phenylacetic acid (PAA). Plants were formed by 83.9-86.0% of explants, they had high weight (234−253 mg), two shoots measuring 2.07−2.37 cm and 5.8−8.3 roots measuring 1.00–1.24 cm. Mean plant weight was lowest on the media with indole-3-butyric acid (IBA) (185–192 mg). Both explant buds formed single shoots, but their development was typically uneven. The differences in the length and weight of shoots were lowest on the media with IAA and at lower PAA concentrations. Plants from the media with IAA and the control medium were distinguished by a higher number of nodes. The percentage share of shoots in the total plant weight was highest on the media with PAA (62.1–62.7%), and lowest at higher concentrations of α-naphthaleneacetic acid (NAA) (47.9 and 48.9%). Parts of explants immersed in media developed callus, and the highest amounts of this tissue were found in the media with NAA. 92.3% of plants survived the acclimatization. The applied procedure may be used for rapid in vitro cloning of selected stevia genotypes. The use of one medium enables reduction of seedling production costs. Moreover, cyclical cloning and extending the production scale is possible.

scholarly journals In Vitro Mass Multiplication and Assessment of Genetic Stability of In Vitro Raised Artemisia absinthium L. Plants Using ISSR and SSAP Molecular Markers

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
B. Kour ◽  
G. Kour ◽  
S. Kaul ◽  
M. K. Dhar
Keyword(s):  
Molecular Markers ◽  
Genetic Stability ◽  
Multiple Shoot ◽  
Nodal Explants ◽  
Direct Regeneration ◽  
Naphthaleneacetic Acid ◽  
Shoot Induction ◽  
Multiple Shoot Induction ◽  
Artemisia Absinthium

The present investigations were made attempting to develop a rapid, reliable, and reproducible in vitro regeneration protocol for Artemisia absinthium L., a medicinal plant of Kashmir Himalayas. Out of several auxin-cytokinin combinations tested, Murashige and Skoog’s (MS) medium supplemented with 0.5 mgL−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mgL−1 kinetin (Kn) was found to be the best for the callus induction. On the other hand, 4.5 mgL−1 6-benzylaminopurine (BAP) and 0.5 mgL−1 1-α-naphthaleneacetic acid (NAA) in the medium resulted in maximum shoot induction from the callus. Similarly, BAP and NAA at a concentration of 1.5 mgL−1 and 0.5 mgL−1, respectively, proved to be the best for the multiple shoot induction from nodal explants. Numerous shoots were obtained from nodal explants after third subculture. In vitro rooting was maximum on medium containing indole-3-butyric acid (IBA) at 0.5 mgL−1. The genetic stability of the in vitro raised plants of Artemisia absinthium was assessed using the intersimple sequence repeat (ISSR) and sequence-specific amplification polymorphism (SSAP) molecular markers. Both markers were able to detect the somaclonal variations in the callus regenerated plants, while no variation was detected in the plants regenerated from the nodal explants. SSAP has been found to be more useful in detection of variability as compared to ISSR molecular marker. The results of present study concluded that the direct regeneration protocol will be useful for the production of true to type plants of this medicinally important plant. This will go a long way in reducing the pressure on the natural populations for the secondary metabolite production, especially for extraction of essential oils.

scholarly journals In vitro organogenesis and plant regeneration of Passiflora xishuangbannaensis, a species with extremely small populations

2021 ◽  
Author(s):  
Yuan-yuan Meng ◽  
Shi-jie Song ◽  
Sven Landrein
Keyword(s):  
Plant Regeneration ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Basal Medium ◽  
Ex Situ ◽  
Naphthaleneacetic Acid ◽  
South American ◽  
South American Species

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species

scholarly journals 319IN VITRO MATURATION OF EQUINE OOCYTES IN A COMPLETELY DEFINED MEDIUM SUPPLEMENTED WITH PROGESTERONE

2004 ◽  
Vol 16 (2) ◽  
pp. 279
Author(s):  
B. Merlo ◽  
E. Iacono ◽  
F. Prati ◽  
G. Mari
Keyword(s):  
Polar Body ◽  
Basal Medium ◽  
Defined Medium ◽  
Chi Square ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Chi Square Test ◽  
The Media ◽  
Cytoplasmic Maturation

A completely defined medium for in vitro maturation (IVM) of equine oocytes has not yet been developed, since most of the media used for IVM are supplemented with serum or BSA. Furthermore, in this species there is no report about the influence of progesterone on maturation, although it has already been used as supplement (500ngmL−1) in EMMI (Maclellan LJ et al., 2001, Theriogenolgy 55, 310 abst). The aims of this study were to develop a completely defined medium for equine oocyte maturation and to investigate the effect of progesterone on nuclear maturation. Equine oocytes were collected by follicular scraping of abattoir-derived ovaries between April and June. The basal medium for maturation was SOFaa supplemented with pFSH-LH 0.1IUmL−1 (Pluset, Laboratorios Calier, Barcelona, Spain), EGF* 50ngmL−1, ITS (Insulin, Transferrin, Sodium selenite), L-cysteine 1.2mM, Maturation SOF (MSOF). Compact cumulus-oocyte complexes were selected, washed three times in H-SOF and matured in one of the following media (15–20 oocytesmL−1): (1) MSOF+FCS 10% (MSOF-FCS), (2) MSOF+progesterone 100ngmL−1 (MSOF-P4), (3) MSOF. After 24h of culture in 5% CO2 in air at 38.5°C, the oocytes were denuded by gently pipetting in a 0.25% trypsin solution, washed and stained with Hoechst 33258 (10μgmL−1 in PBS) for 30min at room temperature. Oocytes were examined under a fluorescent microscope to assess nuclear maturation. Only oocytes with an evident polar body and metaphase II plate (MII) were considered mature. The experiment was done in 6 replicates. Chi Square test was used for statistical analysis (Statistica for Windows – Stat Soft Inc., Tusla, OK, USA). Significance was assessed for P<0.05. The results of this study show that MSOF can be considered a suitable completely defined medium for IVM of equine oocytes. Adding progesterone significantly (P<0.05) increases the nuclear maturation rate at 24h of culture. It can be speculated that although cumuls cells produce this hormone, supplementation is useful to reach progesterone concentrations similar to those present in follicular fluid (early dominant 63.4±19.3ngmL−1, healthy preovulatory follicle 1094.3±170.9ngmL−1; Gerard N et al., 2002, Reproduction 124, 241–248). Further studies are needed to investigate the influence of progesterone on cytoplasmic maturation and to test the effect of different progesterone concentrations and time of maturation in a completely defined system.*All chemicals were purchased from Sigma, St. Louis, MO, USA, unless otherwise stated. Table 1 Maturation of equine oocytes in different media

In vitro mass propagation of Piper betle L

2019 ◽  
Vol 48 (3) ◽  
pp. 559-566
Author(s):  
Salim Khan ◽  
Barna Goswami ◽  
Shahina Akter ◽  
Mousona Islam ◽  
Afsana Huq Noon ◽  
...  
Keyword(s):  
Nodal Explants ◽  
Direct Regeneration ◽  
Leaf Segment ◽  
Piper Betle ◽  
Nodal Segment ◽  
Root Induction ◽  
Regeneration System ◽  
Petiole Explants ◽  
Best Response

An efficient in vitro regeneration system was developed for Piper betle L. through direct and indirect organogenesis from nodal segment, leaf segment and petiole explants. Highest direct regeneration was recorded when nodal explants were cultured on MS with 1.0 mg/l BAP and 1.0 mg/l Kn where 80% explants produced multiple shoots and the average number of shoots per explants were 3.20. Remarkable results on callus induction and shoot initiation were observed when the explants cultured on MS + 2.0 mg/l BAP + 0.5 mg/l Kn + 1.0 mg/l IAA. It was observed that nodal explants were showed best response on shoot/explants 13.2 ± 4.5 after 8 weeks of callus culture on MS medium with 0.5 mg/l BAP. The best response towards root induction was observed on half strength of MS with 0.25 mg/l IBA. The well rooted plants were successfully acclimatized and transferred to soil.

Genetic transformation of Stella De Oro daylily by particle bombardment

2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou
Keyword(s):  
Southern Blotting ◽  
Basal Medium ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Semi Solid ◽  
Independent Transformation

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis

In vitro propagation of Crambe maritima

1987 ◽  
Vol 65 (1) ◽  
pp. 72-75 ◽  
Author(s):  
J. Y. Peron ◽  
E. Regnier
Keyword(s):  
In Vitro Propagation ◽  
Indoleacetic Acid ◽  
Adventitious Shoots ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Petiole Explants ◽  
Acclimatization Period

A method for rapid micropropagation of sea kale (Crambe maritima L.) was developed. Petiole explants placed in vitro on a medium containing 0.5 mg/L indoleacetic acid (IAA), 6.0 mg/L kinetin, and 1.5 mg/L benzylaminopurine developed callus within 15 days and shoots within 28 days. Nearly four adventitious shoots could be developed within 3 weeks by placing the initial shoot on media without IAA. To develop roots, the shoots were then transferred to the basal medium containing 0.1 to 1.0 mg/L indolbutyric or α-naphthaleneacetic acid. Rooted plantlets were obtained within 2 or 3 weeks. After an acclimatization period of 6 weeks in a greenhouse in unsterilized medium, the plantlets could be set outdoors.

scholarly journals Total Polyphenol Content, in vitro Antifungal and Antioxidant Activities of Callus Cultures from Inula crithmoides

2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Anahi Bucchini ◽  
Laura Giamperi ◽  
Donata Ricci
Keyword(s):  
Methanol Extract ◽  
Antioxidant Activities ◽  
Leaf Explants ◽  
Basal Medium ◽  
Alternaria Solani ◽  
Antimicrobial Activities ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Stable Free Radical

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.

scholarly journals Micropropagation of Clerodendrum phlomidis L.F.

2016 ◽  
Vol 24 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Mafatlal M. Kher ◽  
Deepak Soner ◽  
Neha Srivastava ◽  
Murugan Nataraj ◽  
Jaime A. Teixeira da Silva
Keyword(s):  
Callus Formation ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Nodal Explants ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoot Proliferation ◽  
Clerodendrum Phlomidis

Abstract Clerodendrum phlomidis L. f. is an important medicinal plant of the Lamiaceae family, particularly its roots, which are used for various therapeutic purposes in a pulverized form. The objective of this study was to develop a standard protocol for axillary shoot proliferation and rooting of C. phlomidis for its propagation and conservation. Nodal explants were inoculated on Murashige and Skoog (MS) medium that was supplemented with one of six cytokinins: 6-benzyladenine, kinetin, thidiazuron, N6-(2-isopentenyl) adenine (2iP), trans-zeatin (Zea) and meta-topolin. Callus induction, which was prolific at all concentrations, formed at the base of nodal explants and hindered shoot multiplication and elongation. To avoid or reduce callus formation with the objective of increasing shoot formation, the same six cytokinins were combined with 4 μM 2,3,5-tri-iodobenzoic acid (TIBA) alone or in combination with 270 μM adenine sulphate (AdS). Nodal explants that were cultured on the medium supplemented with 9.12 μM Zea, 4 μM TIBA and 270 μM AdS produced significantly more and longer shoots than on medium without TIBA and AdS. Half-strength MS medium supplemented with 8.05 μM α-naphthaleneacetic acid was the best medium for root formation. Most (75%) in vitro rooted plantlets were successfully acclimatized under natural conditions.

scholarly journals THE USE OF NANO-SILVER FOR DISINFECTION OF Pennisetum alopecuroides PLANT MATERIAL FOR TISSUE CULTURE

2019 ◽  
Vol 18 (3) ◽  
Author(s):  
Marzena Parzymies ◽  
Krystyna Pudelska ◽  
Monika Poniewozik
Keyword(s):  
Tissue Culture ◽  
Plant Material ◽  
Nodal Explants ◽  
Grass Species ◽  
Silver Particles ◽  
Ag Nps ◽  
Nano Silver ◽  
The Media ◽  
Pennisetum Alopecuroides

Initiation of tissue culture of many plant species is a very difficult stage due to appearance of many contaminations. The other problem might be a choice of media for regeneration. Initiation of grass species tissue cultures are thought to be very difficult. Therefore, a research was undertaken to evaluate the use of nano-silver particles for plant material disinfection and to estimate a medium Pennisetum alopecuroides. The plant material were buds and nodal explants that were disinfected in 2% NaOCl for 30 min or 0.1% HgCl2 for 1 min. Half of the explants disinfected with NaOCl were soaked in 50, 100 or 250 mg·dm Ag–3 NPs for 1 hour. Explants not soaked in nano-silver were placed on media with Ag NPs at concentrations of 4, 8 or 16 mg·dm–3. An influence of growth regulators on Pennisetum alopecuroides was evaluated in vitro. Regenerated shoots were placed on MS media with: 3 mg·dm–3 BA + 0.3 mg·dm–3 IBA, 3 mg·dm–3 KIN + 0.3 mg·dm–3 IAA, 1 mg·dm–3 BA + 0.1 mg·dm–3 IBA. It was observed that the use of nano-silver particles lowered the level of contamination. The best results were obtained when Ag NPs was used at concentration of 100–250 mg·dm–3 alone or as a supplementation of the media, at concentration of 4 mg·dm–3 for nodes and 16 mg·dm–3 for adventitious buds. The use of nodal explants allowed to obtain less contamination. Regeneration depended on a media content. The most regenerated shoots were obtained on the MS media supplemented with 1 mg·dm–3 BA and 0.1 mg·dm–3 IBA.

scholarly journals Alkaloids of in vitro biomass of papaver nudicaule l

2018 ◽  
Vol 25 (03) ◽  
pp. 90-96
Author(s):  
Otgonpurev S ◽  
Munguntsooj B ◽  
Ariunjargal B ◽  
Munkhtsetseg D
Keyword(s):  
Thin Layer ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Proliferative Capacity ◽  
Whole Plant ◽  
Murashige Skoog ◽  
Layer Chromatography

Papaver nudicaule  L has a long history as a being useful for the treatment of many diseases in Asian and European countries. Aim of this study was to cultivate callus and cell suspension culture in vitro using plant phytohormones. The proliferative capacity was tested on shoot cultivated on Murashige-Skoog (MS) basal medium testing auxins: -Naphthaleneacetic acid (NAA)  in combination with cytokinine: 6-Benzylaminopurine. We determined production of alkaloids by four-week old callus and cell suspension biomass of Papaver nudicaule using thin layer chromatography comparing standard sanguianarine. This result displayed us the easier approach to isolate pure one alkaloid from the biomass that those whole- plant. Нүцгэн намуу ургамлын in vitro биомасс дахь алкалоид Хураангуй: Нүцгэн намуу (Papaver nudicaule) нь эмийн болон чимэглэлийн ач холбогдолтой ургамал юм. Уг ургамлын үрээс каллусын болон эсийн суспензийн биомассыг гарган авахдаа нафтален цууны хүчил болон бензил аденин өсөлтийн бодисын хослол бүхий тэжээлт орчинд өсгөвөрлөн өсөлтийн параметрүүдийг тогтоолоо. Ургамлын in vitro биомасст нимгэн үеийн хроматографийн аргаар сангвинарин алкалоидыг стандарт бодистой харьцуулан тодорхойлов. Сангвинарин нь ургамлын эд эрхтэний төрөлжилтөөс үл хамааран үүсдэг биологийн өндөр идэвхтэй хоёрдогч нийлэгжилтийн бүтээгдэхүүн юм. Түлхүүр үг: өсөлтийн гормоны хослол, каллусын эд, суспензийн нягт, нимгэн үеийн хроматографи

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