Assessment of bioscompatibility of isocyanurate-containing polyurethanes with ifosphamide by in vivo and in vitro methods.

Background. The creation of polymeric composite materials with pronounced biological activity, which are able to act as implants with local prolonged action of the immobilized substance can be widely used in medical practice. Objective. Study of cellular reactions of surrounding tissues of experimental animals to implantation of polymeric composite materials based on isocyanurate-containing polyurethanes with ifosfamide, study of histotoxicity of the obtained materials by tissue culture method. Methods. Polymeric composite materials based on isocyanate-containing polyurethanes without and with ifosfamide were implanted into the body of white laboratory Wistar rats. Cellular responses of the organism and possible changes in the structure of test specimens after implantation were studied by light microscopy by analysis of histological micropreparations. In order to study the peculiarities of the dynamics of growth and development of fibroblastic elements, the method of tissue culture was used. Results. Conducted biological studies by in vivo and in vitro methods allowed to evaluate the effect of immobilized ifosfamide in the structure of isocyanurate-containing polyurethanes on cellular reactions of surrounding tissues during implantation in experimental animals, as well as the effect of extracts from developed polymer samples on cultured cell growth. Conclusion. It was found that the implantation of isocyanurate-containing polyurethanes with ifosfamide led to the development of intense cellular reactions in the area of implant placement, primarily the reaction of round cell elements. The presence of ifosfamide in the structure of the polymeric implantation material probably affected the proliferation of cellular elements, inhibited regenerative processes in the early stages of the study and delayed the formation of a mature connective tissue capsule around the implanted samples. The tissue culture method showed that when making an extract of isocyanurate-containing polyurethanes with ifosfamide in the culture medium, there was a large variability of cell forms, which led to the appearance of macrophage-like elements.

Microtubers production by using Temporary Immersion System (TIS) bioreactor to potato varieties

As the national potatoes growth has become lower from year to year, it is now known that the problems came from the lack of certified potato seed varieties and the minimum access to sophisticated technology to make a good potato variety. The solution that can be made is to utilize microtubers as an efficient factor. The purpose of this research is to find the most effective method in micro tubers cultivation in vitro by comparing the conventional tissue culture method and the Temporary Immersion System (TIS) bioreactor method to four different potato varieties (Granola L., Dayang Sumbi, Atlantic Malang, Maglia). This research uses a split-plot design with a completely randomized design by using two factorial. The result of this study shows that the microtubers in the multiplication and production step using the TIS bioreactor method has a higher average compared to the conventional tissue culture method. As the various details, Dayang Sumbi has the highest parameter such as most sprouts, primer roots, diameter, wet weights, and fastest time growth. Granola L. excel in planlet height and most tubers. Atlantic Malang in the most multiplication and nodus. Meanwhile Maglia excel in most leaves.

PENGARUH VARIASI ZAT PENGATUR TUMBUH IAA, BAP, KINETIN TERHADAP METABOLIT SEKUNDER KALUS SIRIH HITAM (Piper betle L. Var Nigra)

AbstrakPiper betle L. var Nigra (sirih hitam) merupakan salah satu jenis tanaman obat yang berpotensi untuk dikembangkan dengan metode kultur jaringan. Penelitian ini bertujuan untuk mengetahui pengaruh variasi zat pengatur tumbuh Indole Acetic Acid (IAA), Benzyl Amino Purine (BAP), Kinetin terhadap senyawa metabolit sekunder yang terdapat kalus sirih hitam. Eksplan daun dari sirih hitam ditanam pada medium Murashige dan Skoog dengan 4 perlakuan kombinasi (I1,0K1,0; I1,0K1,5; I1B1,5; I0,5B0,5). Kalus dipelihara selama 8 minggu. Simplisia kalus sirih hitam di maserasi menggunakan pelarut metanol dan diidentifikasi menggunakan Gas Chromatography Mass Spectra (GCMS). Berdasarkan analisis GCMS menunjukkan bahwa senyawa metabolit sekunder dengan presentase tertinggi terdapat pada masing- masing perlakuan yaitu senyawa gamma sitosterol (14,88%) pada perlakuan I1,0K1,0 ; senyawa 14-Beta H Pregna (15,94%) pada perlakuan I1,0K1,5 serta senyawa beta-d-glucopyranoside (15,54%) pada perlakuan I0,5B0,5; dan senyawa beta-d-glucopyranoside (5,63%) pada perlakuan I1,0 B1,5. Katakunci: Kalus, Piper betle L. var Nigra, senyawa metabolit sekunder AbstractPiper betle L. Var Nigra (black betel) is one type of medicinal plant that has the potential to be developed by tissue culture method. This study was aims to determine the effect of variations in growth regulator Indole Acetic Acid (IAA), Benzyl Amino Purine (BAP), and Kinetin on secondary metabolite compounds contained in black betel callus. The explant of black betel’s leaf was cultured on Murashige and Skoog medium with four combination treatments (I1.0K1.0; I1.0K1.5; I1B1.5; I0.5B0.5). The callus was maintained for eight weeks. Simplisia of black betel’s callus was macerated using methanol solvent and identified using Gas Chromatography Mass Spectra (GCMS). GCMS analysis showed that secondary metabolites with the highest percentage in each treatment were gamma sitosterol compound (14.88%) in treatment I1.0K1.0; 14-Beta H Pregna compound (15.94%) in I1.0K1.5 treatment and beta-d-glucopyranoside compound (15.54%) in I0.5B0.5 treatment; and beta-d-glucopyranoside compound (5.63%) in I1.0 B1.5 treatment.Keywords: Callus, Piper betle L. Var Nigra, secondary metabolite compounds

Usage of the tissue culture method for the development of new forms of Fagopyrum esculentum Moench

The article presents results of usage of the tissue culture method on buckwheat (Fagopyrum esculentum Moench) together with the selective factor increased concentrations of heavy metal ions in the nutrient medium. On the basis of Izumrud (Russia), Kitavase (Japan), hybrid Izumrud x Kitavase there were developed regenerant lines tolerant to copper and zinc. Negative action effect of copper ions was observed from the second generation in the second posterity of the plants-regenerants, developed from the callus, in the form of the chlorophyll-free mutants. Positive effect of the heavy metals was observed on increasing of rutin content of the plants-regenerants in vitro. Selective evaluation in the field conditions of the samples tolerant to heavy metal let to choose from them the most adaptive to monsoon conditions in Primorskykrai, which combine valuable traits of big grain (weight of 1000 grains 36-38 g), high seed productivity (1,9-3,2 g) and high rutin content in aboveground mass of plants (2,8-4,3 %).

The Effect of Gibberellic Acid (GA3) On The In Vitro Seed Germination of Manggosteen (Garcinia mangostana)

Mangosteen is considered as fruit of many benefits with great popularity, rendering it valuable as an export commodity. Unfortunately, it has a relatively slow growth and long early fruiting time. Generative propagation of mangosteen by seed result in low-yielding productionof qualified seedswith recalcitrant nature which means it cannot be stored for a long time. Whereas the result of vegetative propagation using grafting, branch cuttings and budding has difficulty rooting out. Tissue culture method requires growth regulators to control organogenesis and morphogenesis so it can accelerate the growth of the mangosteen explants. This study was arrangedintwo- factor completely randomized design (CRD). The observed variables werethe strength of the medium (½ and 1 MS) and GA3 concentrations (0 ppm; 0.25 ppm; 0.5 ppm; 0.75 ppm and 1 ppm). The variatonson GA3 concentration (0 ppm; 0.25 ppm; 0.5 ppm; 0.75 ppm; 1 ppm) on different strength MS medium (½ MS and 1 MS) did not significantly affect the growth of radicle and sooth in the in vitro seeds germination of Mangosteen (Garcinia mangostana).

Genotypic variability of Pinus sylvestris L. on the drought-resistance attribute

There was an increase and intensification of droughts in connection with global climate change in recent decades. Not all plant organisms are able to adapt to changing environmental conditions. Therefore, the question of stressresistant (drought-resistant) genotypes selection for breeding is quite urgent. This problem also concerns forest tree plants, including Scots pine, which is one of the main foresters in the Voronezh region. The results of vegetative and generative sphere analysis of individual Scotch pine trees with the help of biotechnology, molecular-genetic and cytogenetic method are given in this study. The possibility of applying the tissue culture method in vitro for testing initial plants for stress resistance, including drought resistance, is explained by the interconnection of cells, tissues and the whole plant properties. It is shown that the cytogenetic characteristics of the seed progeny and the indicators of callusogenic reactions do not always coincide in different genotypes: in some cases energy resources are spent on ontogeny protection, while, in other cases, to reproductive function maintain. There are trees, in which the state the generative sphere in arid years is at the level of optimal years, and their callus cultures reaction remains unchanged even under simulated conditions of drought. Based on the results obtained for the selection of Scots pine drought-resistant genotypes we suggest applying a system of criteria characterizing both the ability of the vegetative sphere to survive in drought conditions on the basis of the tissue culture method in vitro (callus tissue formation speed, its viability, frequency of callusogenesis), and the state of the generative sphere with the help of cytogenetic analysis of seed progeny (frequency of mitosis pathologies, the proportion of cells with micronuclei, mitotic activity). The expediency of applying the biotechnological approach was proved by analysis of the gene expression level of stress proteins: the level of AbaH gene expression correlates greatly with the indicator of the viability of callus cultures, including ones on a nutrient medium with an additional stress agent (NaCl). Trees that can be assigned to drought-resistant ones, according to the results of the analysis, should be recommended for use in breeding.

Comparative Study of In Vitro Root and Shoot Proliferation from the Node Explants of Asparagus racemosus Willd

Asparagus racemosus Willd. locally known as Kurilo or Shatavari is found throughout the tropical and subtropical regions of Nepal. It is a high value herb because of its medicinal and nutritional values which is leading this species decrease in its natural habitat. Hence, to conserve this species, an in vitro tissue culture method of its multiplication has been applied as an effective method of ex situ conservation. In the experiment, although most of the treatments induced highly insignificant number of adventitious shoots, low concentration of IBA (0.1-0.5 mg/l) in combination with relatively higher concentrations of Kinetin (1.0-2.0 mg/l) in MS medium were found to be significantly inducing up to 8.33±1.308 shoots/node. Among the treatments where hormones were used singly, IAA 0.5 and Kinetin 1.0 induced 4.83±1.08 and 4.66±1.43 shots/node respectively. Hence, a protocol for the effective multiplication of this species has been developed which can be used accordingly.

Induction of Indirect Somatic Embryogenesis on Embryonic Axis of TRI2025 Tea Clone

Tea (Camellia sinensis L.) is classified as cross-pollinated crop and vegetative multiplication becomes commercially the main method of propagation with some limitations such as high heterogeneity and poor in survival rate and also in rooting. A proven tissue culture method, somatic embryogenesis, is the only challenging way to meet the needs of tea seedlings in large quantities. The study was conducted with TRI2025 tea clone selected from Polyclonal garden of PT. Pagilaran (Batang, Central Java). The explants were cultured on MS media supplemented with four concentrations of 2,4-D (0, 1, 2, and 5 mg L-1) in two incubation conditions; dark and light. The results showed the only concentration of 2,4-D that can induce somatic embryo was 2 mg L-1 2,4-D in light condition and its percentage was about 5%. Other concentrations of 2,4-D that given for treatments both in two conditions will not induce somatic embryo. This study needs more improvements for getting powerful and efficient of method to get somatic embryo-derived plant and also for futher successful genetic engineering of tea biotechnology.

In vitro Cultivation of Newly Reported Wild Edible Mushroom Volvariella bomybycina from Nepal

Wild edible mushrooms are becoming endangered all over the world. Very few wild edible mushrooms are found in natural habitat. Volvariella bombycina is an edible and medicinal mushroom. The mushroom was collected in natural habitat growing on Populus tree. Mycelium of the mushroom was developed in PDA slant tubes by tissue culture method, incubated at 25°C for 1-2 weeks. Spawn was developed in wheat grains after incubation at 25°C for 2-3 weeks. Substrates were formulated for the development of fruiting bodies by combination of paddy straw, saw dust and rice husk. Fruiting bodies of V. bombycina was cultivated in these substrates after incubation at 28 ± 2°C for 2-4 weeks. The work describes the optimized process for in vitro culture of wild edible mushroom Volvariella bomybycina.Nepal Journal of Biotechnology. Dec. 2017 Vol. 5, No. 1: 27-31