Adaptation of microclones of blackberries to in vivo conditions

The results of studying the effect of mineral fertilizing on rhizogenesis and the development of aboveground organs of regenerant plants of blackberry thornless adaptable to in vivo conditions in the laboratory of selection, vegetable growing and horticulture, cloning “UNITS” Agrotechnopark “of Belgorod State Agrarian University are presented. Regenerated plants of thornless blackberry cultivar Agavam were adapted to in vivo conditions in a peat-perlite mixture with the addition of microelements and growth regulator root 16 days earlier than in the control. An active growth of the aboveground part and roots of regenerated plants of thornless blackberry was noted on the 21st day, in the control - on the 42nd day after the start of adaptation. By the end of the rooting stage on the 24th day, the regenerant plants formed an aerial part of two pairs of leaves 22 mm high and a developed root system - 37 mm. The mineral and hormonal composition of nutrient media for the cultivation of thornless blackberries has been optimized, an effective combination of physical and chemical factors at different stages of micropropagation has been determined, which enhance the proliferation of shoots and roots, and the dependence of the efficiency of adaptation of regenerated plants to in vivo conditions has been established. Along with traditional breeding methods, new opportunities for solving the problem of thorn-free blackberry varieties are provided, along with traditional breeding methods, which make it possible to accelerate the process of obtaining valuable planting material to provide the population and the processing industry with valuable berry products.

An Analysis of the Micropropagation and Peculiarities of Eggplant (Solanum melongena L.) Cultivation Technology

This review is devoted to biotechnological methods and approaches used in eggplant selection. Using in-vitro techniques, micropropagation makes it possible to obtain Solanum melongena L. plants identical to the original ones. This article discusses various aspects of eggplant microcloning; draws conclusions about the successful induction of shoots, which is an important stage required for the regeneration of the callus; and describes the methodological issues of origin, explant treatment, cultivation features, composition of nutrient media, growth regulators, growing conditions and rooting algorithms, and adaptation of the resulting regenerant plants. The microclonal reproduction of Solanum melongena L. by organogenesis from cotyledon and hypocotyl explants is described. After explants were introduced into an in-vitro culture, significant differences in the nature of growth and development of the tissues in the nutrient media were observed on the 6-8th day. A comparative analysis of the stimulating effect of growth regulators revealed that thidiazuron with a concentration of 0.02 mg/l had the highest efficiency; its effect on the regenerative capacity of the explant depended on the concentration in the medium. The dependence of all stages of microclonal reproduction on the genotype is shown. Keywords: Solanum melongena L., eggplant, nightshade, microcloning, biotechnology

Effect of rhizosphere microorganisms on the adaptation of regenerated plants of apple clonal rootstocks to ex vitro conditions

In a vegetation experiment, the effect of bacterial preparations Extrasol, Fitosporin-M based on bacterial strains Bacillus subtilis and bacteria strains of the genus Pseudomonas – P. chlororaphis OV17, P. protegens 38a, P. putida О9-10 on the number of rhizosphere microorganisms, growth and development of regenerant plants of apple clone rootstocks in ex vitro conditions was studied. After 90 days of growing plants, the greatest number was found in the P. protegens 38a strain – 0.56 million CFU/g roots. Artificial inoculation of the roots of regenerant plants contributed to a significant increase in the total number of native microorganisms as compared to the control. The most diverse bacterial population in terms of cultural and morphological characteristics was revealed in the variant with the treatment of plants with the P. putida O9-10 strain. The introduced bacterial strains contributed to an increase in the adaptive capacity and had a phytostimulating effect on the development of plants. The plant survival rate in the process of adaptation to ex vitro conditions is most influenced by the P. putida 09-10 strain compared to the control. The bacterial preparation Fitosporin-M contributes to the improvement of biometric parameters of plant growth.

ASSESSMENT OF THE INTEGRATED CHITINASE GENE STABILITY IN WHEAT LINES AFTER BIOBALLISTIC TRANSFORMATION

The synthesis of PR-proteins (pathogenesis related proteins), the most studied of which are chitinases and β-1,3-glucanases, occurs in response to infection with pathogens in plants. Information about the exact role of individual PR proteins within plant immunity makes it possible to use certain specific antifungal proteins for the development of transgenic plants with increased resistance to fungal diseases. At the same time, it is important not only to obtain a plant with the desired trait, but also to fix in it the stable expression of the transferred gene and the inheritance of the acquired trait in generations. Herein we have studied the stability of the chitinase gene insertion in T1, T2 and T3 generations of transformed wheat lines obtained by the method of cis-gene transfer. Primary transformed regenerant plants were obtained as a result of bioballistic transformation of the chitinase gene into immature wheat germ of Saratovskaya 29 and Kazakhstanskaya 19 varieties. Following screening of regenerant plants by PCR for the presence of the target gene made it possible to select 6 lines presumably carrying the insert based on variety Saratovskaya 29 and 2 lines based on variety Kazakhstanskaya 19. The seed material of the selected regenerant plants was cultivated in soil conditions and the seeds of the T1 generation were obtained. DNA amplification of 8 selected lines St-29№25, St -29№43, St-29№44, St-29№33, St-29№26, St-29№35, Кz-19№1, Кz-19№2 with specific primers revealed insert-carrying lines, partially cleaved lines and lines with a high degree of insert instability. According to the results of the T2 and T3 generations PCR analysis, a complete absence of insertion in the St-29№35 and St-29№33 lines was revealed, a partial cleavage of the trait in the St-29№43 and Kz-19№2 lines was revealed, and the stable inheritance of the chitinase gene in four lines St-29№25, St-29№44, St-29№26 and Kz-19№1.was confirmed. These lines were selected as promising for in-depth study of their resistance to fungal diseases and further replication.

Assessment of different ways of improving promising potato varieties and hybrids

The results of application of the apical meristem method for the selection of regenerant plants free of pathogens are presented. The study was carried out in laboratory and field conditions of Kemerovo region (2016–2019). The objects of research were a new variety of Pamyati Anoshkinoi and promising hybrids 6-14-11, 22103-10. The experiment on the health improvement of potatoes was carried out by the following methods: 1) the use of chemotherapy with the addition of virus inhibitors to the nutrient medium on the samples containing a viral infection in a latent form; 2) study of vegetative plants of different ages healthy from pathogens using apical and axillary buds as explants. Antiviral drugs and their concentrations were identified: virazole 0.01% + chitosan 0.05% + interferon 0.05% (a combination of drugs) and cycloferon 0.05%, completely suppressing viral infection SBK and MBK in the Pamyati Anoshkinoi variety and 22103-10 hybrid, and YBK in 6-14-11 hybrid. The use of these drugs makes it possible to obtain from 10 to 50% of viable meristems free from viral infections. Research into the improvement of vegetative plants of potato variety Pamyati Anoshkinoi and 22103-10 hybrid revealed phases of growth and development with less accumulation of viral and fungal infections – a period of plant regrowth of 15–20 cm and a phase of flowering. Isolation of the apical meristem from the apical and axillary buds of healthy vegetative plants in these phases of development has a positive effect on the survival of the meristems (3050%), and makes it possible to obtain from 30.0 to 43.3% of regenerant plants free from infections, which ensures a reliable yield in vitro material and reduces the period for obtaining healthy lines by 6.9 times.

BIOTECHNOLOGICAL METHODS - A TOOL FOR CREATING NEW FLAX GENOTYPES RESISTANT TO ANTHRACNOSE

The purpose of this research was to create new flax genotypes resistant to anthracnose using biotechnological techniques and methods. As a result of studies with using a culture of immature embryos and a selective medium, flax regenerant plants resistant to the culture filtrate (CF) of the fungus - anthracnose pathogen Colletotrichum lini Manns et Bolley and line 21 resistant to this pathogen were obtained. A scheme for differentiating flax genotypes in vitro by resistance to anthracnose has been developed. It was established that upon cultivation of immature embryos on Sh-2 medium containing CF of the fungus, the causative agent of anthracnose, of a mixture of strains 680, 677 * 674, 674 * at a concentration of 36.0 ml / l, the amount of morphogenic callus formed in the first and second passages, expressed in percent, and the indicator of the field resistance of this genotype to anthracnosis on an artificial infectious-provocative background was close in value, and the number of morphogenic callus formed in the first and second passages can be used to judge the resistance of the studied genotypes to ant cancer and differentiate them by resistance to this pathogen. The influence of the flax genotype on the potency of cells to morphogenesis under selective conditions was revealed. Cells of genotypes L 957-8-7, Alexim, Punjab, Zaryanka had high morphogenetic activity. The morphogenetic potential of genotypes L 1506-8-4, Rosinka has already been exhausted by 2 ... 3 passages. It was found that biotechnological methods: cell selection in vitro, embryo culture are effective in creating genotypes of flax, more resistant to anthracnose than original forms.

Optimization of core phases of biolistic transformation of potato

We have established the optimal exposure time for internodes and callus plant material of Aksor potato variety on the OSS osmotic medium before and after biolistics and the type of plant explant that is most suitable for that. The optimal objects were explants from internodes of test plants. The best time for exposure of plant material to the osmotic medium before bombardment is 24 hours, and after biolistics – 48 hours. Callus tissue developed on the MS medium with zeatin two weeks later in internodes that were cultivated before bombarding on the OSS for 24 hours and we observed an increase in the callus mass by the end of fourth week. Not a single callus was obtained with rhizogenesis on the R4 medium from internodes and potato calli that were on the osmotic medium within 24 and 72 hours after biolistics. The regenerative capacity of internodes after cultivation on an osmotic medium for 48 hours after bombarding was higher than that of calli. We obtained 12 regenerant plants out of 200 internode explants and 4 regenerant plants out of 150 calli that were used in the experiment.

Production of drought- resistant regenerant plants of spring soft wheat

Climate change is one of the reasons influences to decline grain quality in Kazakhstan. This includes the growing threat of droughts and frosts during the growing season of wheat, increased winds, and climate contrast against the background of soil degradation (salinization, desertification) because of irrational use and disruption of the ecological balance in the biocenosis. The aim of this work is to obtain drought - resistant regenerant plants of spring soft wheat in in vitro culture and their seeds. The authors have introduced 10 domestic varieties of spring wheat into in vitro culture. Obtained morphogenetic callus of wheat have been cultivated on nutrient medium MS, with ttreatment of optimal concentrations of selective agents: polyethylene glycol 6000 - 5%; mannitol - 2%; NaCl – 0,5%. There have been determined the regenerative capacity of wheat genotypes. High morphogenetic potential was observed among 10 cultivars using 2% mannitol and 5% polyethylene glycol, which made up to 44,2% and 38,9%, respectively and characterizes there in vitro adaptive ability to drought. Seeds (R0-R1) of obtained wheat regenerant lines were produced. Structural analysis has been carried out for obtained regenerant plants from selective agents such as mannitol 2%, NaCl 0.5% and polyethylene glycol 5%. As a result, drought-resistant regenerant plants have been selected and grown to produce their seeds (R1).