Анализ структурно-функциональной организации хлоропластного генома карельской березы на основании данных высокопроизводительного секвенирования

The sequencing and annotation of the curly birch chloroplast genome were carried out. A high level of similarity of the structural and functional organization of cpDNA among the species of the Betulaceae family was revealed. A set of primers was developed to assess the level of expression of EST markers of the curly birch cpDNA by the real time PCR method.Проведено секвенирование и аннотация хлоропластного генома карельской березы. Выявлен высокий уровень сходства структурно-функциональной организации хпДНК среди видов семейства Betulaceae. Разработан набор праймеров для оценки уровня экспрессии EST-маркеров хпДНК карельской березы методом ПЦР-РВ

Морфолого-культуральные и молекулярно-генетические особенности коллекционных штаммов микоризных грибов Phialocephala fortinii и Pezicula sp.

A metagenomic analysis of the endophytic microflora of Vaccinium corymbosum L. and Vaccinium myrtillus L. root systems was carried out. Two dominant species of micromycetes forming ericoid mycorrhiza were identified - Phialocephala fortinii C. J. K. Wang & H. E. Wilcox and Pezicula sp. Tul. & C. Tul. Pure cultures of mycorrhizal fungi were prepared, a comprehensive morphological and genetic assay of the strains was carried out. Based on the results of genetic-taxonomic analysis, the assumption of the polyphyletic origin of species belonging to Phialocephala and Pezicula is confirmed.Проведен метагеномный анализ эндофитной микрофлоры корневых систем Vaccinium corymbosum L. и Vaccinium myrtillus L. Идентифицированы два доминирующих вида микромицетов, формирующих эрикоидную микоризу - Phialocephala fortinii C. J. K. Wang & H. E. Wilcox и Pezicula sp. Tul. & C. Tul. Созданы чистые культуры микоризных грибов, проведена комплексная морфологическая и генетическая оценка штаммов. На основании результатов генетико-таксономического анализа подтверждено предположение о полифилетическом происхождении видов, относящихся к Phialocephala и Pezicula

Геногеография ели европейской (Picea abies (L.) Karst.) по данным анализа цитоплазматической ДНК

Norway spruce (Picea abies (L.) Karst.) is one of the main forest-forming species in Belarus. It plays important economic, ecological and social roles. The spruce forest of the region is characterized by a complex history of the formation of its population genetic structure. The aim of this study was the genogeographic analysis of P. abies populations and the description of regional features of its gene pool in Belarus. Molecular genetic analysis of microsatellite (SSR) loci of chloroplast DNA and mt15-D02 locus of mitochondrial DNA of Norway spruce was carried out for samples from 57 naturally originated forest stands. We identified 19 allelic variants of the Pt63718, Pt26081, Pt71936 cpDNA loci and two allelic variants of mt15-D02 mtDNA. The geographical distribution of the alleles has been described and the regional features of the genogeographic differentiation of the spruce forests have been considered. The southern border of the continuous distribution area of P. abies and its island localities lie in the south of Belarus (Brest and Gomel regions). The frequency of occurrence of individual allelic variants of cpDNA SSR loci has the most pronounced deviations from the average values for the whole country in those regions. Analysis of cpDNA showed the presence of certain regional features of the genogeographic structure of the spruce forest in the “south-north” and “west-east” directions. According to mitochondrial DNA analysis higher concentration of P. abies trees of southern (Carpathian) origin is observed in the southwest of Belarus. On the contrary northern (Boreal) origin dominates in the rest of the country. The results of performed genogeographic analysis of Norway spruce populations can serve as a basis for improving the forest seed zoning of the tree species.Ель европейская (Picea abies (L.) Karst.) является одним из основных лесообразующих видов Беларуси, выполняя важные экономические, экологические и социальные функции. Еловая формация региона характеризуется сложной историей формирования своей популяционно-генетической структуры. Целью данной работы являлся геногеографический анализ популяций P. abies и установление региональных особенностей ее генофонда на территории Беларуси. Проведен молекулярно-генетический анализ микросателлитных (SSR) локусов хлоропластной ДНК и локуса mt15-D02 митохондриальной ДНК P. abies в 57 древостоях естественного происхождения. Выявлено 19 аллельных вариантов локусов Pt63718, Pt26081, Pt71936 хпДНК и два аллельных варианта mt15-D02 мтДНК. Установлено географическое распространение аллелей и рассмотрены региональные особенности геногеографической дифференциации еловой формации. Для еловых древостоев юга Беларуси (Брестская и Гомельская области), где пролегает южная граница сплошной области распространения P. abies и ее островные местонахождения, частота встречаемости отдельных аллельных вариантов SSR-локусов хпДНК имеет наиболее выраженные отклонения от их средних значений по стране. Анализ хпДНК показал наличие определенных региональных особенностей геногеографической структуры еловой формации в направлении «юг–север» и «запад–восток», а мтДНК – концентрацию на юго-западе Беларуси деревьев P. abies южного (карпатского) происхождения, на остальной части страны – северного (бореального). Результаты проведенного геногеографического анализа популяций P. abies могут служить основой для совершенствования лесосеменного районирования данного вида

Территориальное распределение аллелей SSR-локусов хлоропластной ДНК сосны обыкновенной (Pinus sylvestris L.) в Беларуси

Scotch pine (Pinus sylvestris L.) is one of the main forest forming species in Belarus, which cover 49.2 % of the country’s forested area. The aim of the study was to investigate spatial distribution of microsatellite (SSR) alleles (chloroplast DNA) and identify the features of the genetic structure and genogeographic differentiation of P. sylvestris populations in Belarus. Molecular genetic analysis of six SSR loci of Scotch pine cpDNA in samples form 73 naturally originated stands was carried out. 35 allelic variants of loci PCP1289, PCP4507, PCP83314, PCP71987, PCP26106, PCP30277 were identified. The analysis of the geographic distribution of the dominant allelic variants showed that the population structure of the pine forest is rather homogeneous. Certain regional differences in a number of cases were found for less common, but also widespread variants. One group of alleles is characterized by an increase or decrease in the frequency of occurrence in the direction from the southwest to the northeast. Another group includes allelic variants which share is maximal in the zone covering the Grodno region, the southwestern part of the Minsk region and the Gomel region, while to southwest and to northeast from this zone their frequency of occurrence decreases or is absent. A number of alleles were found only in one of the analyzed stands or in a limited area, which may indicate their local origin as a result of spontaneous mutations. The obtained results are important both from a general biological point of view in studying the evolution and formation of the genetic structure of P. sylvestris in Belarus, and from a practical point of view, since they allow to improve the forest seed zoning of the species.Cосна обыкновенная (Pinus sylvestris L.) является одним из основных лесообразующих видов Беларуси, древостои которой занимают 49,2 % лесопокрытой площади страны. Целью исследования являлось выявление на основе изучения территориального распределения аллелей микросателлитных (SSR) локусов хлоропластной ДНК особенностей генетической структуры и геногеографической дифференциации популяций P. sylvestris в Беларуси. Проведен молекулярно-генетический анализ шести SSR-локусов хпДНК сосны обыкновенной в 73 древостоях естественного происхождения. Выявлено 35 аллельных вариантов локусов PCP1289, PCP4507, PCP83314, PCP71987, PCP26106, PCP30277. Анализ географического распространения аллельных вариантов показал, что по доминирующим вариантам популяционная структура сосновой формации достаточно однородна. Определенные региональные отличия в ряде случаев обнаружены по менее встречающимся, но также широко распространенным вариантам. Одна группа аллелей характеризуется увеличением или уменьшением частоты встречаемости в направлении с юго-запада на северо-восток. Другая группа включает аллельные варианты, долевое участие которых максимально в зоне, охватывающей Гродненскую область, юго-западную часть Минской области и Гомельскую область, в то время как юго-западнее или северо-восточнее от этой зоны отмечается снижение их частоты встречаемости или отсутствие. Ряд аллелей встречался только в одном из проанализированных древостоев или же на ограниченной территории, что может свидетельствовать об их местном происхождении в результате спонтанных мутаций. Полученные результаты важны как с общебиологической точки зрения при изучении эволюции и процессов формирования генетической структуры P. sylvestris в Беларуси, так и с практической, поскольку позволяют совершенствовать лесосеменное районирование вида

Изучение видового состава микробиома фитофагов цветочных растений на основании данных ДНК-штрихкодирования

Based on the DNA-barcoding data, a molecular-genetic identification of the dominant microbiome species composition of the perennial floral plants phytophages was carried out. Various variants of species combinations in the microbiota–phytophagous system, as well as the ways of phytophagous transmission of pathogenic microflora have been identified.С использованием метода баркодирования ДНК проведена молекулярно-генетическая идентификация доминирующего видового состава микробиома фитофагов многолетних цветочных растений. Выявлены различные варианты видовых сочетаний в системе «микробиота–фитофаг», а также изучены способы переноса фитофагами патогенной микрофлоры

Genetic diversity and structure of the Norway spruce Picea abies (L.) H. Karst. seed orchards of the first and the second order in Belarus

The genetic diversity of 5 seed orchards of the first (I) and 12 the second (II) orders of the Norway spruce Picea abies (L.) H. Karst. was analyzed on basis of the isoenzyme analysis method, was carried the study with 18 isoenzyme genes. Allelic frequencies of occurrence of the isoenzyme loci were established and the basic indicators of genetic diversity of seed orchards of Norway spruce were calculated (proportion of polymorphic loci, number of alleles per locus, mean heterozygosity) and their comparison with the average stock of genetic diversity of spruce stands of natural origin was conducted. Seed orchards I and II orders are not inferior to natural populations by the proportion of polymorphic loci P95 (0.39, 0.44 and 0.44, respectively). In the case of indicator P99, most seed orchards are comparable to natural stands, however, values of P99 for seed orchards I and II orders as a whole are significantly lower than in natural populations (0.50, 0.50 and 0.67, respectively). Range of allelic diversity in seed orchards is higher than in stands of Norway spruce of production forests. However, many allelic variants in the analyzed totality of seed orchards had a frequency of less than 1 %, although they were found in the spruce formation of Belarus with a frequency of 1 to 5 %. The average values of heterozygosity in seed orchards I order are significantly lower (0.127 and 0.131, respectively) with those in stands of production forests of Belarus (0.147 and 0.150, respectively); in seed orchards II order vice versa – are significantly higher (0.163 and 0.162, respectively). The obtained values of the inbreeding coefficients FIS and FIT for seed orchards I and II orders indicate that they are in equilibrium according to Hardy-Weinberg. The genetic structure of seed orchards I and II orders is characterized by homogeneity because coefficients FST and GST have low values (not higher 0.011 and 0.012, respectively). Assessment of degree of genetic differentiation revealed a similarity of the genetic structures of the studied seed orchards and spruce stands of natural origin of production forests

Early detection and identification of the main fungal pathogens for resistance evaluation of new genotypes of forest trees

Growing importance of forest plantations increases the demand for phytopathogen resistant forest trees. This study describes an effective method for the early detection and identification of the main fungal phytopathogens in planting material of silver birch (Betula pendula) and downy birch (B. pubescens) based on the estimation of size of the internal transcribed spacers (ITS1 and ITS2) in the 18S-5.8S-28S rDNA gene cluster, which are species-specific for most micromycetes. Electrophoretic assay of the ITS1 and ITS2 loci allowed us to identify the predominant phytopathogenic fungal species in downy and silver birch in planta. This new molecular genetic method can be used to screen birch and other forest trees for different fungal pathogens in evaluation of disease resistance. This information can be useful in breeding of new genotypes of forest trees including transgenic clones with modified wood composition

The Variability of Cytoplasmic DNA Haplotypes of Norway Spruce in the Provenance Trials

A genetic analysis of mitochondrial (mtDNA) and chloroplast (cpDNA) genomes of 27 climatypes of Norway spruce was conducted in the provenance (Cherikov forestry district, Republic of Belarus). It was found that the analyzed climatypes presented boreal and carpathian mtDNA haplotypes. The first of them is found in all regions of growth climatypes (Priuralsky, Northwest, Central, Baltic, Byelorussian, Southwest), the second – only in Belarusian and Southwest regions. It is shown that polymorphism cpDNA significantly higher in comparison with the mtDNA. For a number of alleles of loci cpDNA the clinal variation was found in the frequency of their occurrence. It was revealed that some cpDNA haplotypes, as haplotypes mtDNA, of Norway spruce can be represented as in all regions of the investigated part of the range as on certain of its territories

Early Detection and Identification of the Main Fungal Pathogens for Resistance Evaluation of New Genotypes of Forest Trees

The growing importance of forest plantations increases the demand for phytopathogen resistant forest trees. This study describes an effective method for early detection and identification of the main fungal phytopathogens in planting material of silver birch (Betula pendula) and downy birch (B. pubescens), based on the estimation of the size of the internal transcribed spacers (ITS1 and ITS2) in the 18S-5.8S-28S rDNA gene cluster, which are species-specific for most micromycetes. The electrophoretic assay of the ITS1 and ITS2 loci has allowed us to identify predominant phytopathogenic fungal species in downy and silver birch in planta. This new molecular genetic method can be used to screen birch and other forest trees for different fungal pathogens to evaluate disease resistance. This information can be useful in breeding new genotypes of forest trees, including transgenic clones with modified wood composition