DNA typing of the gastric mucosa of patients with chronic duodenal ulcer

The work involved a molecular biological technique (ISSR-PCR) using ISSR-primer S-2, with structure (AGC) 6G. Changes in the gastric mucosa in chronic duodenal ulcer disease against the background of severe chronic atrophic gastritis have been analyzed. Noteworthy is the fact that there is a strong correlation between the degree of dysplasia of the epithelium of the gastric mucosa and the mitotic index, the Pearson's correlation coefficient rxy was 0.853, respectively. A strong and very strong correlation relationship between the indicators of the degree of dysplasia of the epithelium of the gastric mucosa by phenotypic characteristics and indicators of DNA typing of samples of the gastric mucosa, the Pearson's correlation coefficient rxy was 0.863, respectively. DNA profiles of the gastric mucosa of patients with duodenal ulcer according to the results of typing by the ISSR-PCR method ranging from 520 to 620 bp. had the character of microsatellite expansions and differed from the profile of the norm, which is evidence of precancerous changes.

Evaluation of The Genotype of Barbus Luteus (Heckel, 1843) in Dhi Qar and Basra by ISSR-PCR Technique

This study was conducted in the Laboratory of Molecular Genetics/Marine Sciences Center/University of Basrah, from 1/1/2019 to 1/1/2020. Tissue samples of Hamri fish were randomly collected from the governorates of Basra (Al-Qurna District) and Dhi Qar (Al-Nasr District). A total samples was 60, including 30 samples from Basra and 30 from Dhi Qar. The results show that the ISSR 12 primer has a genetic morphology 100%, gave the molecular weights in Dhi Qar Governorate (73 and 255) and in Basra Governorate (66, 65). Primer No. (2) also took shape in the province of Dhi Qar (166, 186, 244), and in the province of Basra (194, 237). The primer (ISSR12) gave 8 bundles, with a molecular weight ranging from (73,255) to samples taken from the Dhi Qar and Basra regions. As for the primer (ISSR8).

ДОСЛІДЖЕННЯ МОЛЕКУЛЯРНО-ГЕНЕТИЧНОГО ПОЛІМОРФІЗМУ РОСЛИН РОДУ MISCANTHUS ISSR-ПРАЙМЕРАМИ

Сучасна селекція біоенергетичних культур спрямована на підвищення вмісту целюлози, сухої речовини, підвищення продуктивності біомаси. Представники видів роду Miscanthus характеризуються різноманітністю, однак джерела походження представників різних видів, а іноді і в межах одного виду, не встановлено. Тому постає необхідність у проведенні досліджень по встановленню філогенетичної приналежності та детермінації наявних селекційних зразків, найбільш цінних для подальшої селекційної роботи. Для виділення ДНК з рослинного матеріалу в наших дослідженнях застосовували стандартний метод екстракції з використанням ЦТАБ. ДНК виділяли з вегетативних органів відібраних матеріалів, індивідуально з кожної рослини. Для аналізу молекулярно-генетичного поліморфізму рослин роду Miscanthus, методом ISSR – PCR, було використано три ISSR-праймера. В результаті ампліфікації отримано 14 локусів, з яких 13 виявились поліморфними, що свідчить про високий рівень поліморфізму. Індекс поліморфності локусу коливався у межах від 0,83 до 0,95. Дослідження рослинних зразків роду Miscanthus з використанням маркерів ISSR 2 та ISSR 4 виявили 100% поліморфізм, оскільки отримані за їх участі 11 локусів були поліморфними. За умов проведення ПЛР з використанням праймера ISSR 1 було отримано три алеля: у M. sinensis виявили один алель, у M. sacchariflorus було виявлено два алелі, для M. giganteus встановлено наявність трьох алелей. Найбільшу кількість поліморфних локусів отримано за використання праймеру ISSR 4 для генотипів видів M. sacchariflorus, M. gigantheus, M. sinensis. Отримали диференціацію представників роду Miscanthus різних видів за різницею в кількості отриманих алелей для кожного представника. Більшу кількість алелей було виявлено для представників групи M. gigantheus, що підтверджує його гібридне походження. Використання даних праймерів для селекційної практики дозволяє провести оцінку генетичного різноманіття наявних видів роду Miscanthus.

Transglutaminase from UV Mutated Bacillus cereus NRC215: Production, Purification, and Characterization

Transglutaminase (EC 2.3.2.13, TGase) recorded the highest activity (0.101 U/ml) in bacterial isolate NRC215. 16S rRNA sequencing revealed that NRC215 was identified as Bacillus cereus NRC215 under accession number MT229271 in the NCBI database. UV irradiation was employed to improve TGase production. Five rifampin (RIF) resistant mutants were only isolated from UV-treated Bacillus cereus NRC215 for three minutes. The best mutant, BCrif5, exhibiting induced rifampin resistance, gave TGase with higher activity (0.148 U/ml). The ISSR PCR technique was employed to detect these new rearrangements resulting from UV mutagenesis between the wild-type strain and its mutants. Moreover, TGase has been purified by three-step procedures resulting in a recovery of 28 and 34.63% for wild and BCrif5 strains, respectively. The optimal purified TGase activity was exhibited at pH 7 for wild strain while the mutant BCrif5 at pH 5.0 and 40 °C for both wild and BCrif5 strains. Bacillus cereus NRC215 TGase was activated by Ba+2 (102.50 and 107.06%), while it was inhibited by Cu+2 (30% and 22.35%) for wild and BCrif5 strains, respectively. It could be concluded that Bacillus cereus NRC215 is a promising strain for TGase production, which is beneficial as a food additive.

CHỌN LỌC IN VITRO MỘT SỐ DÒNG CÚC ĐẠI ĐÓA (Chrysanthemum × morifolium) CHỊU MẶN

Cây hoa cúc là một trong những cây được trồng rộng rãi và cũng là một trong những loại hoa cây cảnh quan trọng nhất trên thế giới. Trong nghiên cứu này, chúng tôi thiết lập hệ thống tái sinh chồi bất định từ mảnh là cây hoa cúc đại đóa, sau đó xác định nồng độ NaCl làm áp lực chọn lọc trực tiếp chồi bất định, đồng thời phân tích sự đa dạng di truyền của các dòng sau chọn lọc được thực hiện bằng ISSR-PCR. Kết quả cho thấy, môi trường phù hợp nhất để tái sinh chồi bất định từ mảnh lá ở cúc đại đóa là MS, đường 30 g/lsucrose, 7 g/l agar, pH 5,8, bổ sung 0,5 mg/lBAP. Sau 5 tuần nuôi cấy, số chồi/mẫu, số lá/chồi và chiều cao chồi lần lượt là 4,37 (chồi/mẫu); 4,25 (lá/chồi) và 1,96 (cm). Môi trường để chọn lọc trực tiếp chồi in vitro chịu mặn là môi trường tái sinh chồi bất định có bổ sung muối NaCl 100 mM. Mồi ISSR 873 [5'-(GACA)4-3'] là phù hợp để phân tích mối quan hệ di truyền các dòng cúc sau chọn lọc. Bằng phân tích ISSR-PCR, năm dòng cúc sau chọn lọc (D1-D5) thể hiện sự đa dạng di truyền. Những dòng cúc này là nguyên liệu tốt cho các phân tích tiếp theo về tính chịu mặn.

Population-genetic structure of the eastern sand lizard Lacerta agilis exigua (Reptiilia, Lacertidae), assessed by ISSR-PCR and IRAP-PCR markers

Впервые описано присутствие участков гомологии в геномах восточной прыткой ящерицы к длинным концевым повторам эндогенных ретровирусов Sabrina и SIRE-1 и выполнен сравнительный анализ спектров продуктов амплификации фрагментов геномной ДНК ящерицы Lacerta agilis exigua, полученных с использованием двух типов ДНК маркеров - фрагментов геномной ДНК, ящериц, фланкированных инвертированными повторами микросателлитных локусов и длинными концевыми повторами эндогенных ретровирусов The presence of homology regions in the long terminal repeats of the endogenous retroviruses Sabrina and SIRE - 1 in the genomes of the eastern sand lizard was described for the first time. A comparative analysis of the spectra of amplification products of genomic DNA fragments of the mentioned lizard species obtained using two types of DNA markers - fragments of genomic DNA, flanked by inverted repeats of microsatellite loci and long terminal repeats of endogenous retroviruses, was performed.

Molecular Genetic Analysis of Pinus sylvestris L. and Pinus sibirica Du Tour Populations in Perm Krai Based on Polymorphism ISSR-PCR markers

DNA polymorphism has been studied, indicators of genetic diversity and genetic structure of 3 populations of Pinus sylvestris L. and 3 populations of Pinus sibirica Du Tour in the Perm Krai have been determined. In the populations of P. sibirica, 102 ISSR-PCR markers were found, of which 88 were polymorphic (P95 = 0.863), and in the populations of P. sylvestris — 113 ISSR-PCR markers, 100 of which were polymorphic (P95 = 0.885). The populations of the two studied species of woody plants are characterized by high genetic diversity. At the same time, in P. sibirica, the indices of genetic diversity were slightly higher (HE = 0.195; ne = 1.335; na = 1.330) than in P. sylvestris (HE = 0.166; ne = 1.268; na = 1.212). The analysis of the genetic structure showed that the coefficient of genetic subdivision (GST) in the two studied species of the genus Pinus are similar and amount to 0.320 in P. sibirica and 0.303 in P. sylvestris. The populations of Siberian pine and Scots pine are characterized by an average degree of genetic differentiation, since the interpopulation component accounts for 32.0% and 30.3% of the genetic diversity of these species, respectively. Using the Mantel test, a high correlation was found between genetic and geographical distances in P. sibirica populations (R2 = 0.6871), while P. sylvestris showed a low correlation (R2 = 0.0649). The data obtained are relevant for the preservation of the gene pools of the studied two species of the genus Pinus in the Perm Krai.

Genetic structure of rainbow trout Oncorhynchus mykiss (Salmoniformes, Salmonidae) from aquaculture by DNA-markers

The rational use of valuable fish species from aquaculture is difficult to implement without knowledge of the state of the genetic structure of local stocks. Different types of DNA markers can be used to achieve the goals of selection and breeding work. The genetic structure of a local stock of rainbow trout (Oncorhynchus mykiss Walbaum, 1792) (Salmoniformes, Salmonidae) farmed in Ukraine was studied using DNA-markers: microsatellite (SSR-markers – simple-sequence repeats-markers) and intermicrosatellite (ISSR – inter-simple sequence repeat). Five fragments of trinucleotide microsatellite motifs with a single anchor nucleotide at the 3'-end were used as a primer for analysis by the ISSR-PCR method. Totally, 85 amplicons were obtained across the five loci, of which 92.9% were polymorphic. The total number of alleles ranged from 10 (marker (ACC)₆G) to 23 (marker (AGC)₆G). The following monomorphic amplicons were determined for the studied local stock of rainbow trout: according to marker (CTC)₆C – 770 and 520 bp bands, for the marker (GAG)₆C – 345, 295 and 260 bp, and for the marker (AGC)₆C – 350 bp. The average number of polymorphic bands per locus was 15.8. The selected ISSR primers had a level of polymorphic information content above the average. The most effective markers for molecular-genetic analysis of rainbow trout were (AGC)₆G and (AGC)₆C according to the percentage of polymorphic bands, marker index, effective multiplex ratio and resolving power. The selected ISSR loci allow the genetic structure of the studied local stock to be characterized using the total and the effective number of alleles per locus (Na and Ne were 1.9 and 1.4, respectively), the Shannon index (average value I was 0.4) and the unbiased expected heterozygosity (mean uHe = 0.3). Microsatellite-based analysis showed features of the genetic structure of the local stock of rainbow trout at six microsatellite loci (OMM 1032, OMM 1077, OMM 1088, Str 15, Str 60, Str 73). Allelic diversity was established and alleles with the highest frequency and most typical for the given stock were identified. The Shannon index and unbiased expected heterozygosity were determined using SSR-markers and were 1.42 and 0.79, respectively. This depicts the complexity of the population structure, a high level of genetic diversity and indicates a high level of heterozygosity of local stock. The “gene pool profile” established as a result of ISSR-PCR in the future will help to differentiate local stocks of rainbow trout in aquaculture of Ukraine. Microsatellite markers provide the ability to determine individual features of genetic variation of local populations and to conduct the management of genetic resources on fish farms.

Genetic diversity of Bemisia tabaci Genn. characterized by analysis of ISSR and cytochrome c oxidase subunit I at Qassim, Saudia Arabia

Problems of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) that increased and escalated in the last 40 years seem to be related to one or more aggressive biotypes that appeared to spread steadily worldwide. As well, some biological characteristics of B. tabaci have led some entomologists to change and multiply their methodology to update with the change in the pest genetic structures. This study is the 1st of its kind in Qassim region in KSA in respect of B. tabaci biotypes. Four identification methods (Squash Silverleaf Symptoms (SSL), cross mating, Cytochrome oxidase subunit I (COI) sequences, and ISSR-PCR analysis) were carried out to determine the biotypes of B. tabaci at Qassim regions. Slight SSL symptoms were observed with varying degrees on squash leaves caused by B. tabaci population at Qassim, KSA. Cross-mating among the populations that have the same or similar genetic structures produced fertilized offspring, females and males with higher sex ratios in favor of females, and produced a higher number of eggs. Whereas, B. tabaci populations that varied greatly in their genetic structures produced unfertilized eggs, which produce males only. In the same trend, ISSR-PCR analysis revealed that B. tabaci populations at Qassim regions varied genetically and gathered into four genetic groups. In conclusion, COI analysis is a perfect tool for classification between biotypes in B. tabaci. Therefore, this study declares that B. tabaci that colonized and infest Qassim horticulture has not the same genetic structures but belonging to B biotypes. It could be named as Bemisia species complex.