Growth Regulators Kinetin Research Articles

BIOLOGICAL FEATURES OF HEDYSARUM ALPINUM L. CELL CULTUREWHEN GROWN IN VITRO

The purpose of the research conducted was to obtain a cell culture of Hedysarum alpicum, as a producer of xanthone mangiferin, by achieving the tasks of obtaining a cell culture of H. alpium growing in deep-cultivation conditions and studying its physiological characteristics. Material and methods. The object of the study was a cell culture of H. alpine obtained from the cotyledon of the germination, strain H. alpinum (C) 2016 of the collection of FSBSI «All-Russian Scientific Research Institute of Medicinal and Aromatic Plants» and the native plant grown on the pharmacopeial site of the Botanical Garden of FSBSI «All-Russian Scientific Research Institute of Medicinal and Aromatic Plants», harvest of 2013. Results. It has been established that growth regulators kinetin and 2.4D stimulate growth processes of H. alpium (C) cell culture 2016 in deep cultivation conditions. The duration of growth is 20–23 days. The maximum increase corresponds to 16 g/l on the 20th day of growth. The maximum consumption of oxygen by the cell culture is 30·104 mol/l·h on the 16th day of growth, when the cells are in the regular growth phase. Conclusion. As a result of the work carried out, a stable cell strain of H. alpium was obtained, growing on the surface of the agarized nutrient medium, which has the potential for the synthesis of mangiferin and work has been carried out on studying its physiological characteristics.

In Vitro Callus Induction of Duroia Macrophylla Huber, Chemical Prospection and Biotechnological Potentialities of Its Extracts

The Amazonian wild tree species Duroia Macrophylla (Rubiaceae) produces triterpenes and alkaloids, some of them showed antituberculosis and antitumoral activities. This paper describes the in vitro establishment and callus induction of Duroia Macrophylla leaf explants, preparation of callus extracts and the chemical analysis to verify the presence of alkaloids and terpenes in it. Young leaf explants were established in vitro with 50% of asepsis, being the best disinfestation method with immersion of 5 h at biocide solution (fungicidal and bactericidal), 3 min in 70% ethanol and 15 min in sodium hypochlorite 2%. Once established, the explants in vitro were induced to callus formation process by adding the growth regulators kinetin and 2,4-dichlorophenoxyacetic acid, on Murashige and Skoog medium. In vitro callus were extracted with hexane and methanol and the presence of secondary metabolites was assessed by thin layer chromatography and 1H-NMR analysis. Preliminary chemical analysis suggests that both callus extract of D. macrophylla did not contain alkaloids, but terpenes. Thus, more studies are needed to induce the production of alkaloids in D. macrophylla callus, where the use of elicitors, or other methods, may induce the production of these metabolites in callus, as alkaloids and terpenes showed antituberculosis and antitumoral activities, what are potential of (social and economics) uses with bioprospection activities. This is the first report of D. macrophylla callus induction and the chemical prospection of its extracts.

Induction of Barangan Banana shoot (Musa acuminata L.) from North Nias through Tissue Culture by giving 2,4-D and Kinetin

The research on induction of banana barangan shoot (Musa acuminata L.) from North Nias through tissue culture by giving 2,4-D and kinetin was conducted in the Laboratory of Tissue Culture University of North Sumatera from May until October 2014. The main objective of this research was to know the explants of banana weevil in apical and basal position with growth regulator 2,4-D with kinetin which are able to induce shoots from callus initials. On this research, the treatment tested for induction of shoots was growth regulators 2,4-D in the concentration 0 mg/L, 1 mg/L, 1,5 mg/L, 2 mg/L and 2,5 mg/L with growth regulators kinetin in the concentration 0 mg/L, 5 mg/L, 6 mg/L, 7 mg/L and 8 mg/L. The research was designed using Completely Randomized Design (CRD) two factorial with repetition of experiment. The results of research showed that the interaction between concentration of 2,5 mg/L 2,4-D with 5 mg/L kinetin was fastest (79 days) forming the shoot derived from explant basal part. The concentration of 2,5 mg/L 2,4-D with 5 mg/L kinetin produced the average number of shoots formed for 3.00 and average number of highest shoots for 1.50 cm. Keywords: Musa acuminata, weevil, initiation, callus, shoot, induction, barangan banana, 2,4-D, kinetin.

Mass multiplication of Pogostemon cablin (Blanco) Benth genotypes and increase of essential oil and patchoulol yield

Patchouli ( Pogostemon cablin) is an aromatic species indigenous to Asia that has been cultivated in many parts of the world for the extraction of essential oils from its leaves. The objective of this study is to develop protocols for plant regeneration and acclimatization from three genotypes of patchouli. For the plant regeneration experiment, a completely randomized factorial design, employing the growth regulators kinetin and indole acetic acid (IAA), was used. The acclimatization used different mixtures of coconut coir dust and vermiculite supplemented with NPK fertilizer and limestone. Direct regeneration of plants was observed for MS medium supplemented with low concentrations (0.5–2.0 mg L −1) of kinetin and IAA alone or in combination for all of the studied genotypes. For genotypes POG014 and POG021, there were 175 and 154 shoots, respectively, obtained per explant with the use of 1.0 mg L −1 kinetin and 0.5 mg L −1 IAA. For genotype POG002, there were 41 shoots obtained per explant in a medium supplemented with 1.0 mg L −1 kinetin. The acclimatization of micropropagated plantlets of all three genotypes showed the best results with the vermiculite substrate plus MS medium salts. Essential oil content and the percentage of patchoulol were higher in plants derived from micropropagation in the three genotypes studied.

Indução de brotação apical em mudas provenientes de sementes e do enraizamento de estacas de mangostãozeiro

With the objective to induce apical bud sprouting in young mangosteen seedlings an experiment was carried out with the application of the plant growth regulators kinetin (0, 100 and 400 mg L-1) and gibberellin GA4+7 (50 mg L-1). To obtain rooted cuttings two other experiments were performed: a) application of indolbutyric acid at the concentrations of 0, 100, 500 and 1000 mg L-1 and b) application of α naphtaleneacetic acid, at 6000 mg L-1. In these trials cutting from orthotropic branches were utilized. The results indicated the possibility of using plant growth regulators to obtain more uniform root stocks in a shorter nursery stage and, despite the low percentage of rooting the planting material obtained from cuttings has a great practical potential, since the size of the rooting cuttings is equivalent to that of one year old seedlings.

Indução de brotação apical em mudas provenientes de sementes e do enraizamento de estacas de mangostãozeiro

With the objective to induce apical bud sprouting in young mangosteen seedlings an experiment was carried out with the application of the plant growth regulators kinetin (0, 100 and 400 mg L-1) and gibberellin GA4+7 (50 mg L-1). To obtain rooted cuttings two other experiments were performed: a) application of indolbutyric acid at the concentrations of 0, 100, 500 and 1000 mg L-1 and b) application of α naphtaleneacetic acid, at 6000 mg L-1. In these trials cutting from orthotropic branches were utilized. The results indicated the possibility of using plant growth regulators to obtain more uniform root stocks in a shorter nursery stage and, despite the low percentage of rooting the planting material obtained from cuttings has a great practical potential, since the size of the rooting cuttings is equivalent to that of one year old seedlings.

EFFECT OF PLANT GROWTH REGULATORS AND ORANGE JUICE ON GROWTH OF CALLUS FROM FRUIT TISSUES OF WASHINGTON NAVEL ORANGE

The effect of the plant growth regulators kinetin, gibberellic acid (GA3), and 2,4- dichlorophenoxyacetic acid (2,4-D), either alone or in combination with orange juice, on the in vitro growth of mesocarp and endocarp explants from 90–120 day-old Washington Navel Citrus sinensis orange fruits has been determined. Both fruit tissues formed callus. From endocarp 100% callus was formed in all cultures, even without growth regulators in the culture medium, while callus growth was lower from mesocarp. The addition of orange juice at a final concentration of 15% (v/v) to the basal medium without growth regulators increased the callus growth, specially that derived from endocarp. However, the high stimulant effect of orange juice observed on callus growth was markedly dependent on the addition of 2,4-dichlorophenoxyacetic acid, maximum callus growth from endocarp or mesocarp tissues being obtained with orange juice 15% (v/v) and 10 μM 2,4-D.

Degradation of polychlorinated biphenyls by hairy root culture of Solanum nigrum

Polychlorinated biphenyls (PCBs) present in the commercial mixture Delor 103 were transformed by hairy root culture of Solanum nigrum. Plant growth regulators kinetin, 2,4-dichlorophenoxy-acetic acid, benzylaminopurin and/or naph-thaleneacetic acid, influenced the cells' growth and transformation of PCBs in a different manner. The cells were able to transform PCBs even if they ceased growing. Young inoculum (16 days) had a 20% lower PCB conversionthan did older inocula (37, 68 days) while biomass increase was much higher using young inoculum. With increasing size of inoculum, transformation of PCBs was stimulated. After 30 days of incubation the average amount of residual PCBs was 40% of the controls at initial PCB concentration of 100 ppm.

Plantlet regeneration via somatic embryogenesis from subcultured callus of mature embryos ofPicea abies (Norway spruce)

Embryogenic callus was initiated from radicles of mature embryos removed from imbibed seeds (24 h). Embryogenic and other nonembryogenic types of callus proliferated on a modified half-strength Murashige-Skoog medium (MS) basal medium (BM) supplemented withmyo-inositol, casein hydrolysate (CH), L-glutamine (gln) and growth regulators kinetin (KN), N6-benzyladenine (BAP) each (20×10−6 M), 2,4-dichlorophenoxyacetic acid (2,4-D) (50×10−6 M) Embryogenic callus bearing suspensor-like cells in a mucilaginous gel matrix was isolated and maintained by subculture every 10 to 12 days on BM with KN, BAP each (2×10−6 M) and 2,4-D (5×10−6 M). Somatic embryos developed spontaneously from the callus on this medium at 23±1° C. Closer examination revealed that numerous polyembryonic clusters, comprised of elongated cells (suspensors) and small dense cells with large nuclei (somatic embryos), occurred in the viscous gel. When this enriched embryonal-suspensor mass was subcultured to low 2,4-D (1×10−6 M), globular embryos developed by 40 to 60 days. Upon transfer to a liquid medium without growth regulators, the embryos elongated and developed cotyledons and shoots with needles. Plantlet development was completed by 30 days in a basal medium without CH, gln and growth regulators. The total culture time was 150 days. Approximately 40±10 embryos were formed from 500 mg of initial callus. Somatic embryogenesis became aberrant if embryos remained attached to the callus mass and were not subcultured within 10 to 12 days according to the described protocol. Somatic embryos were encapsulated in an alginate gel and stored at 4° C for nearly two months without visible adverse effects on viability.

Factors that influence dormancy in milkweed seeds

Seeds of Asclepias syriaca L. (milkweed) have an inherent dormancy and these seeds could be forced to germinate by various treatments including stratification, cutting or removal of the seed coat, gases, alternating temperature, and a number of chemicals. Of the various treatments, the growth regulators kinetin and gibberellic acid were most effective in overcoming dormancy and a combination of these two had a synergistic effect. The increased germination percentage after 5 weeks of stratification was not accompanied by an increase in endogenous gibberellin levels. Treatment of the seeds with abscisic acid after stratification resulted in a complete inhibition of germination whereas treatment with kinetin resulted in an increase in germination. It is concluded that dormancy in milkweed seeds could be broken by many factors and regulated by an intricate interplay between growth promoters and inhibitors.

Effects of growth regulators on fruit set and June drop of pears and apples.

To increase yields of pear and apple trees the growth regulators kinetin, gibberellin (A3 and a mixture of A4 and A7), Alar and CCC were applied by spraying on blossoms or fruitlets and by injection into the trunk. Kinetin and particularly gibberellins were effective in stimulating fruit set in pears. Doyenne du Cornice pear set more fruits with a mixture of gibberellin A4 and A7 than with gibberellin A3. Although the majority of stimulated fruits abscised during the June drop period, the crop was markedly increased. The growth retard-ants Alar and CCC were inactive in increasing fruit set, but prevented excessive drop of fruits when applied after a gibberellin application.-Lab. TuinbPl, Wageningen. (Abstract retrieved from CAB Abstracts by CABI’s permission)

Ribonucleic acid synthesizing activity of spinach chloroplasts and nuclei

Purified chloroplast and nuclei preparations from spinach leaves possess RNA polymerase activity, as measured by the incorporation of 3H-ATP into an acid-insoluble product. In both types of organelle the polymerase reaction is dependent on the presence of all four nucleoside triphosphates, Mg ++, and an ATP-generating system. The reaction is inhibited by RNase, DNase, and actinomycin D. No stimulation was found with spermidine, with ammonium sulfate, or with the plant growth regulators kinetin, indoleacetic acid, and gibberellic acid. RNA polymerase activity was higher in leaf extracts from young plants. The product of the in vitro RNA polymerase reaction was isolated by phenol extraction and sucrose density gradient centrifugation. The chloroplast product had a major peak at about 11S, and was somewhat polydisperse in the range 11–23S. The product of the in vitro nuclear reaction had a maximum at about 8S and was less polydisperse than the chloroplast product. Short-term incorporation of 3H-uridine by intact spinach plants revealed the presence of a rapidly labelled RNA fraction in the chloroplasts with sedimentation properties similar to the product of the in vitro chloroplast polymerase reaction. The corresponding rapidly labelled RNA fraction in the nuclei resembled the in vitro nuclear product but was more polydisperse in the high molecular weight regions of the gradient. In long-term, in vivo incorporation experiments the labelled RNA pattern of both chloroplasts and nuclei corresponded closely to the ribosomal and transfer RNA fractions.