Optimization of biotechnological process clonal micropropagation in vitro of Asparagus officinalis L.

The study presents the results of obtaining regenerated plants of asparagus from seeds. Surface sterilizing the seeds by 0,75% sodium hypochlorite for 30 min is effective, during this obtained 83% viable sterile plants. The Murashige and Skoog medium supplemented with 6‑benzylaminopurine (2 mg/L), inositol (100 mg/L) and thiamine (0,4 mg/L) was found to be the best for seed germination. The expediency of using kinetin (1 mg/L) as a growth regulator to obtain a homogeneous plant material was established. The reproduction coefficient was 6,0. Only 11% of the explants formed callus. For the selection needs and production of somaclonal variants, the use of the culture medium with indole-3-acetic acid (0,2 mg/L) and 6‑benzylaminopurine (1 mg/L) is justified. In this condition reproduction coefficient was 3,7, and the level of different intensity callusogenesis was 59%. The rooting of obtained plants was performed in Murashige and Skoog medium supplemented with a half dose of macro- and micronutrients and growth regulators. Rooting frequency was up to 63%. The knowledge of hormonal requirements helps to promote isolated tissue and cells technologies of asparagus with purpose of rapid propagation and obtaining healthy, high-quality planting material.

Optimation of Auxin and Cytokinin on Enhanced Quality and Weight of Coffea liberica Somatic Embryos

Coffea liberica is a variety of coffee that tolerant to marginal land, especially peatlands. One of propagation methods in C. liberica is somatic embryogenesis(SE) which producing large number of true-to-type plant seedlings in a short time. This research aimed at studying the effect of application of plant growthregulator (PGR) on quality and weight of somatic embryo of C. liberica. Somatic embryo in development stage was induced by Murashige and Skoog medium containing cytokinin as benzyl amino purin (BAP) and auxin as 2,4-dichlorophe-noxyacetic acid (2,4-D). While cotyledonary embryo in germination stage was induced by Murashige and Skoog medium containing cytokinin (BAP) and auxins as 2,4-D, indole acetic acid (IAA) and naphthaleneacetic acid (NAA). The resultsshowed that the application of auxins and cytokinins on development stage affected the formation of embryos, texture of calli, color of calli and embryos, and weight of somatic embryo. It also influenced the shoot and root formation, color and weight of geminating embryos of C. liberica at the germinating stage. During the development stage, addition of 1 mg/L BAP in the absence of 2,4-D in MS medium produced the highest quality of somatic embryo of C. liberica. This medium also produced heaviest somatic embryos but with lighter callus. While in germination stage, all medium treatments produced a typical germinating embryo. Coffea liberica germinating embryo growth optimally on MS medium containing 0.5 mg/L BAP as a single chemical or 0.5 mg/L BAP in combination with 0.5 mg/L IAA for shooting development. Whereas on rooting development, addition of 0.5 mg/L NAA on MS medium produced an optimal germinating embryo. Moreover, germination embryo of C. liberica recorded the highest in terms of dry weight on MS media with addition of 0.5 mg/L BAP. Application of appropriate concentration of auxin and cytokinin is needed to support the formation of somatic embryo and germinating embryo.

Effect of the Different Concentrations of Protective-stimulating Complex (HFC) on the Growth and Development of Grape Seedlings in the Tissue Culture at the Different Nutrient Levels

Background: Humic substances plays a vital role in the plant tissue culture as a growth hormone for in vitro propagation of many plant seedlings. The aim of this study was to investigate the effect of added humic-fulvate complex (HFC) at the various concentrations on the growth and development of grape seedlings in in vitro at the different nutrient levels. Methods: The cutting of khasansky grape were cultivated on ¼ Murashige and Skoog medium or ½ Murashige and Skoog medium either alone or supplemented with the humic-fulvate complex at the different concentrations at (0.1, 1 and 10 ml/l). Then, they were cultured for 4 weeks under a controlled environment. Result: The data observed that the low concentration of Murashige and Skoog medium (¼ MS) for in vitro rooting of grape cv. ‘Khasansky’ either alone or combined with HFC at the various concentrations significantly increased the rooting percentages and the total length of roots and stimulating the rate of vegetative growth compared with cultivated in ½ MS medium either alone or with supplemented with HFC. ¼ MS+ 10 ml/l HFC was the best treatment for improving the growth of khasansky grape seedlings.

INDUCTION OF ABACA BANANA ROOTS BY IN VITRO USING KINDS OF MEDIA AND THIAMIN

Abaca is a type a banana with high economic value with it stem fiber used in textile and paper industries. As a superior commodity, its number is relatively limited, with the need of a largeplanting area to meet the high market demand. The aim of the research was to observe the abaca banana explants response to various media and Thiamin. The experiment was done at Biotechnology laboratory, UPN “Veteran” Yogyakarta. Treatments were arranged in completely randomized design with 2 factor. The first factor is the growing media: Murashige & Skoog, a half Murashige & Skoog media, Vacint & Went Media and the second factor is the Thiamin concentration: 2 mg/L; 3 mg/L; 4 mg/mL.The results showed there is an interactions on the parameters of planlet height, number of lenghth of root in the combination of Murashige and Skoog and thiamin 3 mg/L medium. Murashige and Skoog medium produced the highest fresh weight and dry weight and Thiamin concentration 3 mg/L produced fresh and dry weight in the highestKey words: abaka, root induction, various media, thiamin

Use of growth inhibitor chlormequat chloride in potato culture in vitro

The publication presents data on the use of growth inhibitor chlormequat chloride in in vitro potato culture on varieties of different maturity groups: Meteor (early), Zekura (mid-early), and Northern Lights (mid-season). Five dosages of the investigational product were studied, ranging from 0.1125 to 1.8 g/l. It was found that in maximum doses of the product there was a strong inhibition of all growth processes in all varieties. The research results showed that the most optimal concentration of chlormequat chloride is 0.225 g per 1 liter of Murashige and Skoog medium. At this dose, on 30th day of cultivation, there was a decrease in the height of microplants from the control by 63.2-85.1%, in the root length - up to 15.0% and their number - up to 22.8% and an increase in the number of internodes by 6.5-22,0 % depending on the variety. The investigational product had an effect on formation of microtubers; in the Meteor variety, their largest number was 89.5% in the nutrient medium with a dose of 1.8 g/l, in the Zekura variety - 93.0% in the nutrient medium with a dose of 0.9 g/l. The new technique makes it possible to lengthen the periods between cuttings of test-tube plants by 2.3 times. This, in turn, reduces the cost of maintaining the in vitro collection material of potatoes in the summer-autumn period, and improves its quality, since each additional cutting cycle affects a more rapid degeneration of the variety. Also, microplants grown with growth inhibitor during subsequent relocation to a standard Murashige and Skoog medium did not show an aftereffect.

Effects of Cytokinin on in vitro Propagation of Bauhinia variegata L.

Mature seeds of Bauhinia variegata L were cultured on half strength Murashige and Skoog medium. For experimentation, nodal cuttings were used as explants from in vitro growing plants. Cytokinin, N-benzyl-9-(2-tetrahydropyranyl) (BPA), kinetin(6-furfurylaminopurine), zeatin, 6-(4-hydroxy-3-methyl-trans -2-butenyl amino purine), 2- isopentenyl amino purine (2-ip), and benzylaminopurine (BAP) were tested for best propagation. Well grown plants were achieved in medium supplemented with 5 µM BPA and 0.5 µM BAP. The propagated plants were acclimatized very well after transferred to the field.

INDUCING SOME SECONDARY METABOLITES FROM CALLUS CULTURES DERIVED FROM Plantago psyllium AND Plantago major EXPOSED TO COBALT STRESS

This experiment was conducted to study the influence of cobalt concentrations on the production of seven flavonoid compounds in callus derived from Plantago psyllium L. and Plantago major L. Results showed that the best combination of 2,4-D and kinetin concentrations add to Muroshige and Skoog medium to obtain the highest fresh weight of 541.0 mg was 3.0 and 1.0 mg.L-1 respectively. psyllium stimulated callus produced the highest fresh weight of 365.7 mg. The addition of 75 ppm of cobalt resulted in a significantly lower fresh weight of P. psyllium callus (139.8 mg). The interaction between Plantago species and cobalt concentrations was significant. The callus inducted from P. major had significant increases of the scutallarein, apigenin, nepetin and luteolin compounds with 26.40, 22.64, 14.93 and 26.20 µg.100mg-1 dry weight, respectively. The production of the hispidulin compound was increased in P. psyllium at 29.40 µg.100mg-1 dry weight. Also, the addition of cobalt metal stimulated the production of flavonoids at 50 ppm cobalt producing the highest amounts of hispidulin and luteolin at 40.30 and 41.60 µg.100mg-1 dry weight, respectively. Meanwhile, 75 ppm cobalt treatment produced the highest amount of scutallarein, apigenin, nepetin and aucubin at 25.61, 23.25, 15.90 and 13.70 µg.100mg-1 dry weight, respectively. The callus inducted from P. major treated with 50 ppm of cobalt showed the highest production of scutallarein, apigenin and luteolin at 30.33, 32.26 and 51.90 µg.100mg-1 dry weight respectively. Baicalein reached 16.46 µg.100mg-1 dry weight, at 75 ppm of cobalt metal treatment in callus inducted from P. psyllium.

Fabrication and properties of calcium pectinate hydrogel nanoparticles with trans-cinnamic acid

Hydrogel negatively charged (–13.5 ± 5.0 mV) calcium pectinate nano- and submicroparticles (50–150 nm) were obtained. A technique for entrapment of a plant growth regulator (trans-cinnamic acid) in the particles up to 40 wt. % has been developed. It has been established that the complete release of trans-cinnamic acid in the Murashige–Skoog medium takes 2.5 hours. The obtained particles of calcium pectinate do not affect the growth processes of cells in suspension culture and can be used as neutral carriers for growth regulators.

Nepenthes mirabilis (Lour.) Druce Planlet at a Various Levels of Murashige & Skoog Medium Density In Vitro

This study aims to determine the variation of the stomata index of the Kantong Semar (Nepenthes mirabilis) planlet at a various medium density of the Murashige and Skoog. This study used a Completely Randomized Design using one factor (medium density of the Murashige and Skoog). We used 5 levels of medium density, i.e.: 1/16 MS, 1/8 MS, 1/4 MS, 1/2 MS, and MS. Homogeneity test used Levene’s test of 5% significance level, then analysis of variance is carried out at 5% significance level and followed by Tukey test at 5% significance level. The results showed that the lower the level of Murashige and Skoog medium density on the Nepenthes mirabilis plantlet, the stomata index also increased.