scholarly journals Induction of Tetraploids from Petiole Explants through Colchicine Treatments inEchinacea purpureaL.

2009 ◽  
Vol 2009 ◽  
pp. 1-7 ◽  
Author(s):  
Dahanayake Nilanthi ◽  
Xiao-Lu Chen ◽  
Fu-Cheng Zhao ◽  
Yue-Sheng Yang ◽  
Hong Wu
Keyword(s):  
Treatment Duration ◽  
Basal Medium ◽  
Echinacea Purpurea ◽  
Naphthaleneacetic Acid ◽  
Root Tips ◽  
Petiole Explants ◽  
Chimeric Plant ◽  
Diploid Cells ◽  
Almost All

Petiole explants were obtained from in vitro grown diploid (2x=22)Echinacea purpureaplantlets. Shoots were regenerated by culturing the explants on MS basal medium containing 0.3 mg/L benzyladenine (BA), 0.01 mg/L naphthaleneacetic acid (NAA) and four concentrations (30, 60, 120, and 240 mg/L) of colchicine for 30 days, or 120 mg/L of colchicine for various durations (7, 14, 21, and 28 days). The regenerated shoots were induced to root on MS basal medium with 0.01 mg/L NAA, and then the root-tips of the regenerated shoots were sampled for count of chromosome number. It was found that a treatment duration of >7 days was necessary for induction of tetraploid (4x=44) shoots, and treatment with 120 mg/L colchicine for 28 days was the most efficient for induction of tetraploids, yielding 23.5% of tetraploids among all the regenerated shoots. Chimeras were observed in almost all the treatments. However, the ratio of tetraploid to diploid cells in a chimeric plant was usually low. In comparison with diploid plants, tetraploid plants in vitro had larger stomata and thicker roots with more root branches, and had prominently shorter inflorescence stalk when mature.

Morphogenesis in Punica granatum (pomegranate)

1986 ◽  
Vol 64 (8) ◽  
pp. 1644-1653 ◽  
Author(s):  
Kanchan Jaidka ◽  
P. N. Mehra
Keyword(s):  
Callus Tissue ◽  
Basal Medium ◽  
Punica Granatum ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Root Tips ◽  
Isolated Roots ◽  
Irregular Form ◽  
Diploid Cells

Explants of root, hypocotyl, cotyledon, stem, shoot tip, and leaf of seedlings obtained by in vitro germination of seeds, as well as embryos excised from seeds, were utilized for the induction of callus. Murashige and Skoog basal medium supplemented with naphthaleneacetic acid (4 ppm), kinetin (2 ppm), and coconut water (15%) was found to be optimal for induction and growth of callus from all explants. Growth rate experiments were performed with callus to study the effect of different growth regulators at various concentrations. The calluses were heterogeneous in nature and consisted predominately of diploid cells, although a few polyploid cells were also observed after two and four subcultures. Plantlets, isolated roots, leaves, and shoots were differentiated in various callus cultures. The root tips and shoot tips of such plantlets revealed only diploid constitution. Embryolike structures were formed in callus on transfer to media containing naphthaleneacetic acid and 6-benzylaminopurine. Embryoid development was traced to a single cell which was invariably isolated from the rest of the callus tissue. This initial divided to form a multicelled structure which later gave rise to a globular, ovoid, or heart-shaped embryoid, or one with irregular form. The embryoids germinated into complete plantlets with root and shoot. The embryoidal initials were mostly diploid but occasional aneuploids or polyploids were observed.

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 Effects of Sucrose and Other Additives on In Vitro Growth and Development of Purple Coneflower (Echinacea purpurea L.)

2014 ◽  
Vol 2014 ◽  
pp. 1-4 ◽  
Author(s):  
Dahanayake Nilanthi ◽  
Yue-Sheng Yang
Keyword(s):  
Plant Growth ◽  
Culture Medium ◽  
Respiratory Infections ◽  
Shoot Multiplication ◽  
Echinacea Purpurea ◽  
Naphthaleneacetic Acid ◽  
Acid Plant ◽  
Purple Coneflower ◽  
Upper Respiratory

Echinacea purpurea (purple coneflower) is being used for the preparation of more than 240 extracts, salves, and tinctures to help cure diseases like rabies, cold, and upper respiratory infections. Hence, efforts were made to develop a culture medium for successful in vitro culturing of cornflower and to regenerate buds and induce roots to enable mass propagation of selected clones. Of the three levels of sucrose tested as a supplement to MS media (Murashige and Skoog’s medium, 1962) 3% showed better rooting of buds and appeared morphologically normal and identical as compared to those grown at higher and lower concentrations (2 and 4%). The additives hydrolyzed lactabumin (0.0, 100, 300, and 900 mgL−1), peptone (0.0, 100, 300, and 900 mgL−1), and yeast (0.0, 100, 300, and 900 mgL−1) to media containing 0.3 mgL−1 BA (6-benzyladenine) and 0.01 mgL−1 NAA (naphthaleneacetic acid-plant growth regulators) has negatively influenced proliferation of shoots. The higher concentrations of the above have delayed the development of plantlets. Shoot multiplication was enhanced by coconut water with 2% being the best among 4 and 8% tested. Shoot organogenesis was not influenced by copper sulphate (0, 1.5, 3, 6, and 12 mgL−1) and silver nitrate (0.0, 0.5, 2.5, and 12.5 mgL−1) supplements and at higher concentrations of the above inhibited plant growth.

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

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

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 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

scholarly journals In vitro Multiple Shoot Regeneration from Petunia hybrida

2019 ◽  
Vol 7 (10) ◽  
pp. 1554
Author(s):  
Rebaz Rasul Habas ◽  
Musa Turker ◽  
Fethi Ahmet Ozdemir
Keyword(s):  
Petunia Hybrida ◽  
Survival Rates ◽  
Basal Medium ◽  
Multiple Shoot ◽  
Shoot Length ◽  
Peat Moss ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Shoot Number

An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.

scholarly journals Plant Regeneration from in Vitro Culture of Embryonic Axis Explants in Common and Tepary Beans

1992 ◽  
Vol 117 (2) ◽  
pp. 332-336 ◽  
Author(s):  
Mohamed F. Mohamed ◽  
Paul E. Read ◽  
Dermot P. Coyne
Keyword(s):  
Plant Regeneration ◽  
Common Bean ◽  
Basal Medium ◽  
Continuous Light ◽  
Embryonic Axis ◽  
Naphthaleneacetic Acid ◽  
Tepary Bean ◽  
Bud Induction ◽  
Multiple Buds

A new in vitro protocol was developed for multiple bud induction and plant regeneration from embryonic axis explants of four common bean (Phaseolus vulgaris L.) and two tepary bean (P. acutifolius A. Gray) lines. The explants were prepared from two embryo sizes, 3 to 4 mm and 5 to 7 mm, corresponding to pods collected after 15 and 25 days from flowering, respectively. The embryonic axis was cultured on Gamborg's B5 basal medium with 0, 5, 10, or 20 μm BA in combinations with 0, 1, or 2 μm NAA. The cultures were maintained at 24 to 25C under continuous light or incubated in darkness for 2 weeks followed by continuous light before transfer to the secondary B5 medium (0 or 2 μm BA or 2 μm BA plus 4 μm GA3). Adventitious roots or a single shoot with roots formed on the explants cultured on media without plant growth regulators. Multiple buds were induced on all BA media, but more were produced with 5 or 10 μm for most lines. Dark incubation greatly enhanced multiple bud initiation. Shoot buds were not produced on media containing NAA alone or in combinations with BA. On the secondary medium, six to eight shoots per explant for common bean and up to 20 shoots per explant from tepary bean were observed after 3 weeks. Mature, fertile plants were produced from these shoots. Chemical names used: benzyladenine (BA); 1-naphthaleneacetic acid (NAA); gibberellic acid (GA3).

scholarly journals Anther Culture of Potato and Molecular Analysis of Anther-derived Plants as Laboratory Exercises for Plant Breeding Courses

1999 ◽  
Vol 9 (4) ◽  
pp. 585-588 ◽  
Author(s):  
Richard E. Veilleux
Keyword(s):  
Plant Breeding ◽  
Anther Culture ◽  
Molecular Analysis ◽  
Simple Procedure ◽  
Extraction Procedure ◽  
Basal Medium ◽  
Root Tips ◽  
Wide Range ◽  
Simple Medium

Anther culture has been one of the most successful techniques for generating haploid plants over a wide range of species. It is a reasonably simple procedure that can be accomplished successfully without sophisticated laboratory facilities; yet, the plants generated through anther culture can be used to demonstrate the application of many modern methods that have direct applicability to plant breeding. Anthers of diploid potato clones that have been selected for competence in anther culture can be cultured in a simple medium to yield androgenic embryos after 5 weeks. Plant regeneration requires an additional 3 to 4 weeks. Regenerated plants should be large enough 2 weeks after transfer to basal medium for ploidy determination by any of three methods depending on available facilities: chromosome counts in root tips; chloroplast counts in stomatal guard cells; or flow cytometry of nuclei released from in vitro plantlets. DNA can be extracted from anther-derived plantlets using a rapid extraction procedure to demonstrate segregation of PCR (polymerase chain reaction)-based markers such as RAPD (randomly amplified polymorphic DNA), RAMPs (randomly amplified microsatellite polymorphisms), or microsatellites. Microsatellite markers that were heterozygous in the anther donor can be used to verify haploidy in anther-derived plants. If an anther culture laboratory is scheduled early in a semester, such molecular analysis can be planned for late in the same semester.

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