scholarly journals Protoplast isolation and culture from different explants of Musa spp. Cau man

2018 ◽  
Vol 1 (6) ◽  
pp. 106-116
Author(s):  
Huong Thanh Tran ◽  
Cuong Quoc Vo
Keyword(s):  
Physiological Activity ◽  
Young Male ◽  
Male Flower ◽  
Protoplast Isolation ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Meristem Culture ◽  
Male Flowers ◽  
2,4 Dichlorophenoxyacetic Acid

In this paper, the roles of type and concentration of enzymes on protoplast isolation from in vitro leaves, multi-scalps (highly proliferating meristem culture), and young male flower of banana cv. cau man were studied. Respiration rate and content of plant hormones of these materials were analysed. Different techniques were used to culture these protoplasts. The development of protoplasts was observed under fluorescence microscope. The highest yield of protoplast (69.5 x 106 protoplasts / g fresh weight) was obtained from young male flowers after 16 hours treatment with 1.5 % cellulase, 0.25 % pectinase and 0.25 % hemicellulase. The combination of hanging drop cell technique (in 6 days), and carrot feeder layer cells in N6PKM medium supplemented with 0.2 mg/L 2,4- dichlorophenoxyacetic acid (2,4-D), 1 mg/L α- naphthalene acetic acid (NAA), and 0.5 mg/L zeatin are suitable for protoplast development. Protoplasts that were isolated from multi-scalps and young male flowers created the wall and divided when cultured by this method. The development of protoplasts from young male flower began with cell walls creation after 4 days, the cell division was after 6 days, and small colonies formation was after 28 days of culture. The differentiation and physiological activity of cells play an important role on quantity and quality of protoplasts, as on the well as protoplast development.

scholarly journals PENGARUH EKSPLAN DAN ZPT TERHADAP PERTUMBUHAN nEPEnTHES ALBOmARGInATA SECARA In VITRO

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.

scholarly journals CALLUS INDUCTION ON BANANA FLOWER’S EXPLANT IN VITRO USING 2,4-DICHLOROPHENOXYACETIC (2,4-D)

2018 ◽  
Vol 22 (2) ◽  
pp. 66
Author(s):  
RINDANG - DWIYANI ◽  
HESTIN - YUSWATI ◽  
UTAMI -
Keyword(s):  
Acetic Acid ◽  
Callus Induction ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Regeneration Medium ◽  
Indirect Organogenesis ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Banana Cultivar

ABSTRACT  The objective of the study was to obtain the best 2,4-D concentration on callus induction of the banana flowers in banana propagation using indirect organogenesis method. Kesuna, local banana cultivar obtained from Sembung Gede, Tabanan was used as explant material. Callus induction was performed using 2,4-Dichlorophenoxyacetic acid with concentration of 0; 0.5; 1.0; 1.5 and 2.0 ppm. Each treatment was represented by 3 bottles and each bottle was planted with 3 explants, so each treatment was represented by 9 explants of banana flowers. The results showed that the concentration of 2.0 ppm 2.4-D induced callus with the fastest time and gave the highest percentage of the explants producing callus. The calluses were subsequently subcultured into regeneration medium using 0.5 mg/L Benzylaminopurine (BAP) and 0.005 mg/L Napthaleneaceticacid (NAA). The calluses were subsequently sub-cultured into a regeneration medium using 0.5 ppm (BAP) and 0.005 ppm Naphthalene acetic acid (NAA) to induce shoots and roots and performed plantlets.   Keywords: 2,4-Dichlorophenoxyacetic acid, banana’s flowers, callus

scholarly journals Smooth leaf (spineless) Red Spanish pineapple (Ananas comosus) propagated in vitro

1969 ◽  
Vol 73 (4) ◽  
pp. 301-311
Author(s):  
Lii J. Liu ◽  
Evelyn Rosa-Márquez ◽  
Enid Lizardi
Keyword(s):  
Callus Formation ◽  
Ananas Comosus ◽  
Dichlorophenoxyacetic Acid ◽  
Meristem Culture ◽  
Standard Medium ◽  
Shoot Differentiation ◽  
Low Concentrations ◽  
One Year ◽  
2,4 Dichlorophenoxyacetic Acid

Some 40,000 plantlets of Red Spanish pineapple [Ananas comosus (L. Merr.)] were produced via meristem culture. Of these, approximately 50% were spineless. Some of these spineless plantlets reversed to spiny leaf. However, the percentage of reversion from spineless to spiny was 14.1% and that from spiny to spineless was 32.7%. Of the 2,318 plantlets examined in the laboratory and greenhouse during a 3- to 4-month period, 72.9% of the spiny Red Spanish pineapple remained spiny and 85.8% of the spineless remained spineless. One year after field planting, the spineless Red Spanish remained largely spineless and initiated flowering and fruit settings the same as the spiny ones. The standard medium for in vitro propagation of Red Spanish pineapple was improved by supplementing Murashige and Skoog's basic formula (MS) with 0.1 mg/L, 2,4- dichlorophenoxyacetic acid (2,4-D) + 0.5 mg/L benzyl adenine (BA). The callus formation was improved by adding to the same MS formula 10 mg/L BA + 4 mg/L naphtalene acetic acid (NAA). Similarly, shoot differentiation was improved by adding low concentrations of hormone (0.1 mg/L NAA) to the Abo El-Nil and Zettler (AZ) medium.

scholarly journals In vitro plant regeneration of Zenia insignis Chun

2018 ◽  
Vol 13 (1) ◽  
pp. 34-41
Author(s):  
Zhou Yu-qing ◽  
Zhang Meng-jie ◽  
Zhang Deng ◽  
Zhang Jun-jie ◽  
Li Jing-jian ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Basal Medium ◽  
Deciduous Tree ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Root Induction ◽  
In Vitro Plant Regeneration ◽  
Morphogenic Callus ◽  
2,4 Dichlorophenoxyacetic Acid

AbstractZenia insignis Chun is a large, fast-growing deciduous tree. In this study, we successfully developed a reliable and efficient protocol for the regeneration of fertile plants via callus induction from leaf segments of young Z. insignis seedlings. The best results were obtained with a medium containing 11.00 μM 6-benzyladenine (6-BA), 1.20 μM indole-3-butytric acid (IBA), and 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D), which yielded morphogenic callus within 2 weeks at a frequency of 62.23%. We tested the effect of IBA alone and in combination with 6-BA on the bud differentiation response of Z. insignis callus. Shoots differentiated normally when cultured on differentiation medium containing 6.00 μM 6-BA and 1.20 μM IBA. Regenerated buds elongated successfully in medium containing 1.20 μM gibberellic acid (GA3). The elongated shoots were then transferred to Murashige and Skoog basal medium supplemented with various combinations of naphthalene acetic acid (NAA) for root induction; well-developed roots were achieved on MS basal medium supplemented with 0.01 μM NAA at a rooting rate of 89.23%. Rooted plantlets were successfully acclimatised to a greenhouse at a survival rate exceeding 90.00%.

scholarly journals Influence of Growth Regulators on Callogenesis and Somatic Embryo Development in Date Palm (Phoenix dactyliferaL.) Sahelian Cultivars

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Djibril Sané ◽  
Frédérique Aberlenc-Bertossi ◽  
Léopold Ibrahima Djitiningo Diatta ◽  
Badara Guèye ◽  
Abdourahman Daher ◽  
...  
Keyword(s):  
Nuclear Dna ◽  
Leaf Explants ◽  
Suspension Cultures ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Benzyl Adenine ◽  
Embryogenic Cell ◽  
Significant Difference ◽  
2,4 Dichlorophenoxyacetic Acid

This study provides a physiological analysis of somatic embryogenesis in four elite cultivars of date palms: Ahmar, Amsekhsi, Tijib, and Amaside, from the initial callogenesis to establishment and proliferation of embryogenic suspension cultures. Somatic embryos development and in vitro plants rooting were also studied. For each step, auxins and cytokinins concentrations were optimised. The primary callogenesis from leaf explants of seedlings appeared highly dependent on genotype. Ahmar (80%) and Amsekhsi (76%) appeared highly callogenic, whereas Tijib (10%) and Amaside (2%) produced low amounts of calluses. 2,4-Dichlorophenoxyacetic acid appeared favorable to the induction of primary callogenesis and its effect was enhanced by the addition of benzyl adenine or adenine sulfate. Secondary friable calli obtained from chopped granular calli were used to initiate embryogenic cell suspensions in media supplied with 2,4-dichlorophenoxyacetic acid. Suspension cultures showed a growth rate of fourfold after four subcultures in presence of 2,4-dichlorophenoxyacetic acid 2 mg/L. Our results showed that a seven-day transitory treatment with benzyl adenine 0,5 mg/L was necessary to optimize embryos development. Naphthalene acetic acid induced the development of primary orthogravitropic roots during embryos germination. The comparison with cytofluorometry of nuclear DNA amounts showed no significant difference in ploidy level between regenerated plants and seedlings.

scholarly journals An Effective Procedure for In Vitro Culture of Eleusine coracana (L.) and Its Application

ISRN Botany ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Alla I. Yemets ◽  
Galina Ya. Bayer ◽  
Yaroslav B. Blume
Keyword(s):  
Finger Millet ◽  
Callus Formation ◽  
Protoplast Isolation ◽  
Callus Cultures ◽  
Eleusine Coracana ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Efficient Induction ◽  
Free Media

Efficient protocols for callus production, plantlet regeneration, protoplast isolation, and micronucleation of finger millet (Eleusine coracana (L.) Gaertn.) were developed. White nodulated calli were formed on medium with N6 macrosalts, MS microsalts, 2.4-dichlorophenoxyacetic acid (2 mg L−1), kinetin (0.4 mg L−1), 1-naphthalene acetic acid (2 mg L−1), and certain additives. It was found that appropriate supplementation leads to formation of numerous shoots. Healthy rooted plantlets formed on hormone-free media. Although different tested additives had no significant effect on percentage of callus formation, it affected callus quality that further dictated plant-forming capacities. Seedlings were better source tissues for protoplasts isolation compared to callus cultures. About protoplasts were isolated from one gram of seedling coleoptyles. Microcolonies were visible after 20–25 days' incubation on KM8p medium supplemented with glutamine (100 mg L−1) and proline (500 mg L−1). Here we also present a procedure of an efficient induction of micronuclei after chlorpropham (10 μM) and cytochalasin-B (20 μM) seedlings treatment with subsequent microprotoplasts isolation. This technique is discussed for the transfer of alien chromosomes and genes from finger millet by microprotoplast-mediated chromosome transfer.

Callogenesis and plant regeneration of sweet orange cv. Washington Navel from floral organ cultures

2020 ◽  
pp. 19-23
Author(s):  
Nebiha Metoui ◽  
Sabrine Nahdi ◽  
Fethia Dhaouadi ◽  
Dorsaf Yahiaoui ◽  
Malika Meziane
Keyword(s):  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Floral Organ ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Washington Navel ◽  
2,4 Dichlorophenoxyacetic Acid

Washington Navel orange (Citrus sinensis L.) can be infected with virus and virus like diseases that affect not only the production but also fruit quality and the plant’s longevity. For viral sanitation, Washington Navel regeneration was investigated in vitro via floral organ culture. Flowers were collected before opening from healthy Washington Navel trees kept under greenhouse. Floral organs (style/stigma and ovary) were cultured on Murashige and Skoog (MS) medium containing various plant growth regulators combinations of naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The highest rate of callogenesis (95%) was obtained from style/stigma explant cultures on MS medium enriched with 3 mgL-1 BAP, which also resulted in 100% rooted plantlets. Ovary cultures did not show any success on the culture medium with various plant growth regulators combinations. The acclimatization success of rooted plantlets by grafting on Citrus volkameriana rootstocks was about 83%. Thus, these results can be used for mass production of disease-free citrus plants and improve sanitation program of the local citrus genotypes in Tunisia.

In Vitro Shoot Regeneration from Callus Derived from Marubakaido Apple Rootstock under Different Aluminium Concentrations

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 460e-460 ◽  
Author(s):  
Marisa F. de Oliveira ◽  
Gerson R. de L. Fortes ◽  
João B. da Silva
Keyword(s):  
Shoot Regeneration ◽  
Dichlorophenoxyacetic Acid ◽  
Apple Rootstock ◽  
Under Five ◽  
Significant Difference ◽  
Previously Treated ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
In Vitro Shoot Regeneration

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.

Characterization of the Reproductive Mode in Guayule In Vitro

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig
Keyword(s):  
Field Experiments ◽  
Reproductive Mode ◽  
Dichlorophenoxyacetic Acid ◽  
Initial Field ◽  
Breeding Lines ◽  
Reproductive Modes ◽  
Desert Southwest ◽  
2,4 Dichlorophenoxyacetic Acid

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.

In vitro regeneration of Persian poppy (Papaver bracteatum)

Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
Sara Rostampour ◽  
Haleh Sohi ◽  
Ali Dehestani
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
High Performance ◽  
High Efficiency ◽  
In Vitro Regeneration ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Papaver Bracteatum ◽  
Callus Proliferation

AbstractPersian poppy (Papaver bracteatum Lindl.) is an important commercial source of medicinal opiates and related compounds. In this research, calli were induced from seeds, roots, cotyledons and hypocotyls of P. bracteatum at a high efficiency. The optimized callus induction media consisted of the Murashige and Skoog (MS) basic media supplemented with 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin and 15 mg/L ascorbic acid. The concentrations of 2,4-D and ascorbic acid were found critical to callus induction and proliferation. Subsequent subcultures resulted in excellent callus proliferation. Ascorbic acid at concentration 15 mg/L increased the callus proliferation significantly. Maximum callus growth was achieved when the explants were incubated at 25°C. MS salts at full strength were found inhibitory for callus induction, while ľ MS salts were found to favor callus induction. Shoot regeneration of calli in vitro was achieved on ľ MS medium containing 0.5 mg/L benzylamine purine and 1.0 mg/L naphthalene acetic acid. Analysis of alkaloid extracts from Persian poppy tissues by high-performance liquid chromatography showed that thebaine accumulated in the tissues of plants. The thebaine alkaloid profile of the Persian poppy is a well-defined model to evaluate the potential for metabolic engineering of thebaine production in P. bracteatum.

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