Application of molecular genetic methods in breeding of small-seeded lentils for suitability for mechanical harvesting

The variability of weather conditions and the high cost of mineral fertilizers lead to the search for alternative approaches to the optimization of existing and the development of new technological techniques for growing agricultural crops, including soybeans. This article presents the results of research on the study and analysis of the regularities of the height formation of soybean plants of the Madison variety and its average daily linear growth depending on the pre-sowing treatment of seeds with preparations based on active strains of nodule bacteria (Bradyrhizobium japonicum), namely Bioinoculant BTU, Rizolain + Rhizosev, Anderiz and foliar top dressing with biological fertilizers Biocomplex BTU, Gumifrend and Helprost soybean. On the basis of the conducted observations and records, it was established that in the conditions of the right-bank forest-steppe on gray forest soils, the studied preparations of biological origin had a direct effect on both the dynamics of plant height and average daily growth. The maximum height of the plants in the experiment was 78.3 ±7.4 cm, in the phase of physiological maturity, was formed on the variant of the experiment where pre-sowing treatment of seeds with the preparation Biokompleks BTU and foliar fertilizing in the phase of the 3rd trifoliate leaf and budding with organo-mineral fertilizer Helprost soybean was carried out ( 2.5 l/ha), which is 13.3 cm or 20.4% more compared to the control. Another important indicator that characterizes the peculiarities and rates of growth and development of soybean plants during the growing season is the average daily linear increments of the stem. The highest average daily linear growth of soybean plants - 0.65 - 0.66 cm/day, was recorded on the variants of the experiment, where soybean seeds were inoculated with Bioinoculant BTU and Anderiz in combination with foliar feeding of organo-mineral fertilizer Helprost soybean. After conducting a correlation-regression analysis, it was noted that there is a strong positive relationship between plant height indicators and the amount of precipitation during the growing season of soybean plants, while the correlation coefficient was r = 0.805.

Purpose. To evaluate the genotypes of alfalfa feed direction under different conditions of moisture and to identify genotypes that would consistently reproduce a high level of economically valuable traits to create new breeding populations. Methods. The research was conducted at the Institute of Irrigated Agriculture of NAAS during 2017‒2020 years in the field for fodder use on two backgrounds of moisture: drip irrigation and natural moisture. In the year of sowing 1‒3 slopes and 3‒4 for the second year of life were carried out by grassland depending on meteorological conditions of the year, genotype, growing conditions. The subject of research were Unitro and Elegy varieties, root system power selections from samples taken in the Askania-Nova Reserve, Rambler, Abayska multi-colored, Siberian 8 and F3‒F5 hybrid populations. Results. Studies have shown that the characteristics of plant height’ and duration of inter-period periods’ are quite variable, they are subject to fluctuations in years of use of grass, slopes and depend on genetic and biological characteristics of alfalfa populations and meteorological conditions. Alfalfa populations differed in the duration of interspecific periods, height and growth intensity in terms of average daily growth depending on the slope and the year of life of the grassland. The duration of inter-period periods, the onset of oblique ripeness of alfalfa depended on the average daily temperature, the duration of sunshine and the length of the day. The calculated correlation coefficients show that the duration of the inter-period periods is strongly inversely related to the average daily temperature (r = -0.93) and weakly dependent on the day length (r = 0.14) and the duration of sunshine for 1 day (r = -0.18), closer to autumn, they decrease, so with each cut the duration of inter-period periods decreases. The longest was the period of germination-beginning of flowering and was 65 days. This is due to the conditions of this period - low average daily temperature (19.2°C), although the duration of sunshine was high and was 751.8 hours, but for the day the sun shone only 11.7 hours. The shortest period was regrowth-beginning of flowering (second slope) and was 43 days at a high average daily temperature (25.7°C) and a long period of sunshine 12.1 hours per day. In the third mowing, the duration of the regrowth period - the last mowing (beginning of budding) was 55 days, but the plants did not reach the mowing maturity, as it was time for the last mowing before the grass enters the winter. During this period, the average daily temperature drops to 19.9°C, day length to 13:32 hours, the duration of sunshine to 9.2 hours per day. The height of plants on the slopes largely depended on meteorological conditions and was maximum in the second slope, reaching 61.3‒67.7 cm. The lowest indicators of the trait were characterized by populations in the first slope with fluctuations from 51.4 to 56.6 cm and the third ‒ 32. 9‒39.7 cm under irrigation. Under conditions of natural moisture, alfalfa plants were significantly inferior in height with fluctuations of 41.1‒48.1 cm in the first slope and high variability from 18.4 to 29.8 cm in the second, with coefficients of variation on slopes V = 4.0 and 10.5, respectively. The largest linear increase in height in the first year of life was characterized by plants in the second slope with an index of 1.43‒1.60 cm/day under irrigation. The average daily increase in the first and third slopes was much lower and was respectively 0.79‒0.87 cm and 0.62‒0.72 cm under irrigation and 0.63‒0.74 cm and 0.43‒0.63 cm in the first and second slopes in conditions of natural moisture. Alfalfa populations did not differ in the length of inter-period periods during the growing season, and they differed mainly in the years of life of grasses. In the grassland of the second year, this difference was observed depending on the slope and meteorological conditions of the year. Thus, in the second year of life with grass from the first slope to the fourth there was an acceleration of the onset of the beginning of flowering phase (oblique maturity). In the first slope it occurred on the 72-nd day, the second ‒ on the 40-th, the third ‒ on the 39-th and the fourth ‒ on the 51-st day. This occurred with a gradual increase in average daily temperature from 12.4°C in the first slope to 23.3 in the third and its decrease to 21.3°C in the fourth slope. In the change in the duration of sunshine, the same pattern is observed: decrease, then a gradual increase with the following decrease: 8.6‒10.5‒11.1‒9.6 hours/day, respectively. The length of the intercostal periods, similar to the first year of life with grass, was inversely related to the average daily temperature (r = -0.982), rainfall (r = -0.644), day length (r = -0.666) and the duration of sunshine in one day (r = -0.961) and the height of plants which largely depended on meteorological conditions, day length (r = 0.778) and precipitation (r = 0.695). Under irrigation conditions, it varied in different populations on slopes from 64.0 to 70.0 cm in the first slope, 68.1–71.4 cm in the second, and acquired a maximum value of 70.6–76.4 cm in the third slope. Subsequently (the fourth slope) the growth of plants slowed down and their height was in the range of 38.7‒49.7 cm. 40.9‒61.5 cm) from slope to slope and only in some genotypes (Elegy, M.g./P.p., L R/H., A.r.d) in the second slope they remain high. A high positive relationship between the average daily linear growth and plant height (r = 0.991). The highest linear growth was characterized by plants in the third slope, ie in July with fluctuations from 1.81 to 1.96 cm per day, smaller growth rates (1.70-1.78 cm / day) differed plants in the second slope under irrigation. Under conditions of natural moisture, the second slope was characterized by intense growth with fluctuations from 1.41 to 1.69 cm per day. Individual populations (Elegy, M.g./ P.P., L R / H., A.r. d.) Maintained high growth or improved its performance in the third slope. In the first slope, alfalfa plants had weak growth with an average daily increase of 0.91-0.97 cm in favorable irrigation conditions and with an increase of 0.84-0.93 cm / day without irrigation, ie they differed slightly on this basis. In the second slope, compared with the first, the plants grew very quickly, their average daily gain ranged from 1.70 to 1.78 cm (under irrigation). Closer to autumn, the growth and development of plants of the fourth slope slowed down, these processes faded, and the average daily gain was only 0.77‒0.97 cm, however, in the conditions of natural moisture alfalfa plants did not form an aboveground mass. Conclusions. Analysis of the results of the research revealed that alfalfa populations differ in height, growth intensity, average daily growth, duration of intermowing periods on slopes, years of grass life and which depend on average daily temperature, duration of sunshine and day length. Valuable genotypes with high rate of linear growth were identified: Elegy variety and populations of M.g./P.p., L R/H., A.r.d. for further selection work.

Salmonella enterica, Campylobacter and Yersinia species, Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes and Clostridium perfringens are the bacterial pathogens constituting the greatest burden of food-borne disease in Finland. Several molecular genetic methods have been applied to diagnose, discriminate and survey these bacteria. PCR, PCR-RFLP and PFGE are the most widely and successfully used. However, these methods are unable to replace conventional and internationally standardised phenotyping. Electronic database libraries of the different genomic profiles will enable continuous surveillance of infections and detection of possible infection clusters at an early stage. Furthermore, whole-genome sequence data have opened up new insights into epidemiological surveillance. Laboratory-based surveillance performed in a timely manner and exploiting adequate methods, and co-operation at local, national and international levels are among the key elements in preventing food-borne diseases. This paper reviews different applications of molecular genetic methods for investigating enteric bacterial pathogens and gives examples of the methods successfully used in diagnostics and epidemiological studies in Finland.

The article highlights the results of research on the influence of different levels and ratios of the amino acids lysine and arginine in compound feed and the growth and feed consumption of rainbow trout fingerling. For research, 6 groups were formed, 1 control and 5 experimental, each with 500 samples. Combined feed, which differed only in the content of lysine and arginine, was distributed 4 times a day at the rate of 3% of the weight of the fish. In the main period lasting 40 days, the level of arginine changed from 2.00 to 3.60% in compound feed against the background of 2 levels of lysine of 3.00 (groups 1–3) and 3.20% (groups 4–6). If the content of lysine in compound feed is taken as 100%, then the content of arginine varied from 62.50 to 120.00%. We determined the mass of trout, calculated absolute, average daily and relative growth, feed consumption per 1 kg of growth. With an increase in the level of arginine from 2.00 to 3.60% against the background of an effective level of lysine - 3.00% in the combined feed of rainbow trout fingerling, the weight of fish increases by 2.47-2.71% (р˂0.05), absolute, average daily and relative increases, respectively, by 3.04–3.24% (р˂0.05); 3.85% (р˂0.05) and 14.52–15.62 (р˂0.05)%, feed costs per 1 kg of growth decreased by 2.91–3.02%. An increase in the level of arginine from 2.00 to 3.60% against the background of an increased level of lysine - 3.20% in the combined feed of rainbow trout this summer led to an increase in the weight of fish by 2.71-4.30% (р˂0.001), an absolute increase - by 3.14–5.14% (р˂0.001), average daily growth – by 3.85–7.69% (р˂0.001), relative growth – by 12.69–26.49 (р˂0.001) %, combined feed costs per 1 kg of growth decreased by 3.02–4.85%. An increase in the level of lysine in the combined feed from 3.00 to 3.20% against the background of the basic level of arginine - 2.00% in the combined feed of trout this summer probably did not affect the weight and intensity of fish growth. In our studies, no antagonism phenomena between lysine and arginine were established. A one-sided increase in the content of lysine in compound feed did not lead to a decrease in the growth of rainbow trout fingerling. The high rate of growth of rainbow trout this summer was obtained with a ratio between lysine and arginine in compound feed of 1.05–1.20:1. The most effective ratio was 1.13:1.

The main task of maral breeding isproduction increase of antlers. Application of molecular genetic methods in maral breeding contributes to increase in reliability and accuracy of assessment of productivity and breeding value of stag deer at an early age. The purpose of the research is to test methods DNA isolation from biological material of marals. The biomaterial was selected fromstag deer of Altai-Sayan breed (blood, antler crumbs, cartilaginous tissue of the auricles). The work on DNA isolation from maral tissue samples was carried out in the bioengineering laboratory at Altai State University (Barnaul). DNA isolation was conducted using the method based on application of Chelex-100TM Resin chelating agent (Bio-Rad, USA), the method based on DNA precipitation Diamond DNA (OOO ABT, Russia) and commercial kit based on magnetic particles AMPure XP (Beckman Coulter, USA). The purification degree of the isolated DNA was assessed by PCR efficiency in reactions of amplification of the genes of cytochrome oxidase 1, cytochrome B of mitochondrial DNA. The highest concentration of maral DNA was established in isolation based on DNA precipitation (Diamond DNA). The concentration of DNA in the solution was 13.20 ng/µl (blood), 11.30 ng/µl (cartilaginous tissue), 5.74 ng/µl (antler crumbs).

Low positive temperatures have a negative impact on all processes occurring at all stages of rice development. This is especially true during the period of seed germination and sprouting, resulting in reduced seed energy and growth strength. The development of genotypes by introducing genes that provide the ability to overcome this effect is a crucial task for breeders. The application of specific marker systems, which allow differentiating breeding samples for the presence/absence of target genes, is currently of great relevance. In this regard, the main purpose of the current study was to select an effective marker system and develop an optimal protocol (parameters) for PCR analysis, which would allow accurately detecting not only the presence of target genes in the analyzed breeding samples, but also their allelic state. In order to identify quantitative loci (QTLs) there were tested three marker systems, among which the only marker system ‘RM 1377’ showed high efficiency in identifying polymorphism between donors. Using it, a segregating F2 population was diagnosed by PCR analysis and a phenotype estimation was carried out in laboratory. Statistical analysis allowed establishing that this marker was co-inherited. On its basis, there was screened experimental pre-breeding material of rice from the laboratory of physiology of the FRC of Rice. There have been identified samples with donor alleles of target genes, which will be further introduced into the breeding process to study them according to a set of morphometric characteristics and economically valuable traits. The introduction and application of these SSR markers in the breeding process will improve the development of basic rice genotypes with better cold resistance during the period of seed germination and sprouting. Sowing of such seeds can be carried out at an earlier time. This will make it possible to obtain sprouts with optimal density and will significantly improve their potential productivity.

The purpose of this study was to determine the enlargement technique and feasibility analysis of the Pacific white shrimp (Litopenaeus vannamei ) enlargement in CV. Rido Sejahtera shrimp ponds. The white shrimp rearing technique used a semi-intensive system with a stocking density of 75.52 tails/m2 and a pond area of 4,154 m2. Technical analysis produced indicators of several growth factors. These factors are an average body weight, daily growth, biomass, size, survival weight and feed conversion ratio. Based on our results, the average body weight was measured to be 31,65 gram/tail while the average Daily Growth, was recorded at 0,14 gram/day,. Biomass was recorded to be 6.290 kg or 15,14 ton/ha while the size was measured to be 31,6. Our results also yield a 75% survival rate while the Feed Conversion Ratio was recorded to be at 1,39. Based on our finding from a one year financial analysis, we concluded that the product indicators of investment costs would be Rp.307.695.400. Production costs would be around Rp.507.271.347 while revenues are around Rp.1.083.800.300. Based on these findings we concluded that profits Rp. 576.078.953, BEP production volume 3.059 kg, BEP production prices Rp. 40.359, B /C ratio 1.13, and payback period 0.53 years .The results of the study based on financial analysis concluded that the Pacific white shrimp (Litopenaeus vannamei) enlargement financial was feasible to operated, because it was profitable and the B/C ratio value was >1.

BackgroundInfluenza A(H1N1)pdm09 virus infection in Norwegian pigs was largely subclinical. This study tested the hypothesis that the infection causes negligible impact on pigs’ growth performance in terms of feed conversion efficiency, daily feed intake, daily growth, age on reaching 100 kg bodyweight and overall feed intake. A sample of 1955 pigs originating from 43 breeding herds was classified into five infection status groups; seronegative pigs (n = 887); seropositive pigs (n = 874); pigs positive for virus at bodyweight between 33 kg and 60 kg (n = 123); pigs positive for virus at bodyweight between 61 kg and 80 kg (n = 34) and pigs positive for virus at bodyweight between 81 kg and 100 kg (n = 37). Each pig had daily recordings of feed intake and bodyweight from 33 kg to 100 kg. Marginal effects of the virus infection on the outcomes were estimated by multi-level linear regression, which accounted for known fixed effects (breed, birthdate, average daily feed intake and growth phase) and random effects (cluster effects of pig and herd).ResultsThe seropositive and virus positive pigs had decreased (P value<0.05) growth performance compared to seronegative pigs even though feed intake was not decreased. Reduced feed conversion efficiency led to lower average daily growth, additional feed requirement and longer time needed to reach the 100 kg bodyweight. The effects were more marked (P value<0.03) in pigs infected at a younger age and lasted a longer period. Despite increased feed intake observed, their growth rates were lower and they took more time to reach 100 kg bodyweight compared to the seronegative pigs.ConclusionOur study rejected the null hypothesis that the virus infection had negligible adverse effects on growth performance of Norwegian pigs.

Purpose. To investigate the influence of different types of mineral nutrition on the spring barley plants height formation and indicators of average daily linear growth after different precursor crops. Methods. Study of the action and interaction of factors: A – precursor; B – levels of mineral nutrition. Gradation of factors 3x3. Threefold repetition of the experiment with the systematic placement of variants was used. The accounting area made up 30 m2 . Precursor crops for spring barley were winter rapeseed, grain corn, sunflower. Soil preparation in the pre-sowing period depended on the precursor crop and was aimed at maximum preservation and accumulation of moisture in the soil and destruction of weeds. Results. The formation of spring barley plant height dynamics under the influence of different levels of mineral nutrition taking into account the precursor crop is highlighted. It was established that the use of N90P60K60 contributed to the intensive growth of spring barley. Growing spring barley after sunflower without fertilizer made it possible to obtain plant height in the phase of full maturity at the level of 50 cm. Application of mineral fertilizers at the rate of N60P60K60 allowed to increase the height parameters to 55 cm, while application of N90P60K60 – to 63 cm. Analysis of average daily linear growth of spring barley plants during the growing season proved the significant influence of the levels of mineral nutrition on the process of growth. The lowest rates of growth were in the variant without fertilizers – 0.60-0.63 cm/day, while with N60P60K60 – 0.63-0.67 cm/day. The highest rates of average daily linear growth were noted under mineral fertilizing with N90P60K60, and depending on the precursor crop, they were 0.70-0.71 cm/day. Conclusions. The positive role of mineral fertilizers, in particular nitrogen fertilizers, in the formation of high growth rates, starting from the tubing phase till the harvesting ripeness one, has been established. The highest height indicators, 63-67 cm, were confirmed by the measurements carried out in the phase of full ripeness on the variants with N90P60K60 use.

Antimicrobial susceptibility testing has traditionally been based on measurements of minimal inhibitory concentrations of antimicrobials. Molecular genetic studies on antimicrobial resistance have produced a great deal of genetic information which can be used for diagnosis of antimicrobial resistance determinants. Bacteria can acquire resistance to macrolides, lincosamides and streptogramin antibiotics by modification of the target site of the drugs, by active efflux of the drugs, and by inactivation of the drugs. The genetic backgrounds of these resistance mechanisms are well known and several molecular methods for detection of resistance determinants have been developed. Outbreaks of multidrug-resistant tuberculosis have focused international attention on the emergence of Mycobacterium tuberculosis strains that are resistant to antimycobacterial agents. Knowledge of the antimycobacterial resistance genetics and progress in molecular methods has made it possible to develop rapid molecular methods for susceptibility testing. This review presents the genetic background of drug resistance and introduces some methods for genotypic susceptibility testing.

Growth-dependent age estimation in herring ( Clupea harengus L.) larvae

The accumulation of knowledge in the process of scientific and technological development does not occur linearly, but in leaps and bounds: periods of relative stagnation are replaced by scientific revolutions. The scientific revolution is the change of explanatory paradigms by the scientific community. A few years ago, the establishment of the taxonomic affiliation of microorganisms by cultivation and counting by methods of classical microbiology was fraught with a number of insurmountable difficulties. The complex composition of the media, the presence of an excess of inhibiting compounds in the composition of the nutrient media, the need to maintain extreme temperatures, the composition of gases and pressure for cultivation, the susceptibility of the culture to the effects of oxygen excluded uncultured microorganisms from the field of view of the researcher. The emergence of molecular genetic methods made it possible to study the diversity of microorganisms, bypassing the previously obligatory stage of cultivation, isolation of a pure culture and comparison with a reference sample. Through the prism of scientific revolutions and changes in scientific paradigms, the differences between the methods of classical microbiology and molecular genetic methods are revealed. The experience of BIOTROF LLC in the application of molecular genetic methods for the identification of microorganisms in livestock and poultry farming is summarized. Based on the use of 16S metagenomics, it was possible to show for the first time that health disorders of farm animals and birds, a decrease in productivity and the duration of economic use in many cases are associated with dysbiotic phenomena in the gastrointestinal tract. Thus, in cows culled due to lactic acidosis, in comparison with clinically healthy animals, inhibition of the growth of bacteria synthesizing cellulases and microorganisms of the order Selenomonadales, capable of fermenting lactic acid to volatile fatty acids, was observed. Based on the calculation of Pearson's correlations, it was shown that an increase in the content of lactobacilli of the family Lactobacillaceae in the intestines of poultry was associated with an increase in meat productivity, and an increase in staphylococci, with a decrease.

Application of molecular genetic methods in the diagnosis and treatment of human diseases is extremely wide due to a huge amount of hereditary information contained in the human genome. Gene diagnostics allows establishing predisposition to diseases, identification of genetic abnormalities and prediction of pathological outcomes. In addition, gene diagnostics also enables prediction of the individual response to treatment in order to achieve the maximum therapeutic effect. Among all molecular genetic methods, polymerase chain reaction (PCR) diagnostics is a leading approach. Technical simplicity, low cost, high sensitivity and reliability of the method have made PCR diagnostics a routine modality for the risk assessment, diagnostics, and monitoring of the treatment efficiency. Here, we consider the application of PCR diagnostics for the abovementioned tasks and talk about the real-life examples of detecting mutations and chromosomal aberrations which may cause a disease. Further, we discuss the prospects of using a semi-quantitative PCR in medical practice and focus on pharmacogenetics as a key component of a personalised therapy. The lecture is aimed primarily at biomedical students and physicians and represents a continuation of the previous lectures published in Fundamental and Clinical Medicin.

The article substantiates the need to study the development of one-year saplings from cuttings of almond willow (Salix triandra). Purpose of research: identification and analysis of seasonal dynamics of daily growth of shoots in S. triandra clones against the background of excessive atmospheric moisture in the form of three showers in 2020. Material and methods. Research methods: selection, structural-morphological, chronobiological. Object of research: model inbred-clone one-year population of S. triandra. The number of clones is 7; the recurrence is 6-fold. Material: growing annual shoots on one-year saplings; each sapling was formed into a single shoot. Results. Under experimental conditions, the rooting capacity of cuttings was 100%. Dimorphic root systems were formed on the mother cuttings of the excavated seedlings. The annual growth of the strongest shoots was 175 ... 215 cm. The daily growth in shoots reached peak values by June 10: the average daily growth was 3.6 cm/day, the maximum — 4.4 ... 4.9 cm/day. After June 10, the daily growth rate decreased unevenly until the end of the growing season. Some shoots completed growth before the beginning of August, and some continued growth until the middle of August. The cyclical nature of the seasonal dynamics of daily growth in the form of three peaks separated by two dips is established. The third peak is formed after the third rainstorm. Discussion and conclusions. Cyclical seasonal dynamics of daily growth of shoots is characterized by a high degree of synchronization of growth processes. Probably, all the shoots are affected by some powerful pulse synchronizer of unknown nature. It is possible that periodic showers may have a modifying effect on the mechanisms that regulate growth processes. The temporal discreteness of shoot morphogenesis is due to the full or partial realization of the seasonal cycle of shoot development. The duration of growth is due to interclonal and intraclonal differences. Probably, the duration of morphogenesis is controlled by both genetic and epigenetic factors.

SummaryIn‐depth knowledge about spatial and temporal variation in microbial diversity and function is needed for a better understanding of ecological and evolutionary responses to global change. In particular, the study of microbial ancient DNA preserved in sediment archives from lakes and oceans can help us to evaluate the responses of aquatic microbes in the past and make predictions about future biodiversity change in those ecosystems. Recent advances in molecular genetic methods applied to the analysis of historically deposited DNA in sediments have not only allowed the taxonomic identification of past aquatic microbial communities but also enabled tracing their evolution and adaptation to episodic disturbances and gradual environmental change. Nevertheless, some challenges remain for scientists to take full advantage of the rapidly developing field of paleo‐genetics, including the limited ability to detect rare taxa and reconstruct complete genomes for evolutionary studies. Here, we provide a brief review of some of the recent advances in the field of environmental paleomicrobiology and discuss remaining challenges related to the application of molecular genetic methods to study microbial diversity, ecology, and evolution in sediment archives. We anticipate that, in the near future, environmental paleomicrobiology will shed new light on the processes of microbial genome evolution and microbial ecosystem responses to quaternary environmental changes at an unprecedented level of detail. This information can, for example, aid geological reconstructions of biogeochemical cycles and predict ecosystem responses to environmental perturbations, including in the context of human‐induced global changes.