Comparison of genetic diversity in two alien plant species

Bunias orientalis and Erigeron annuus are two alien herbal plant species invading native ecosystems in Lithuania. Both species have similar introduction and invasion history but differ in their life history traits and strategies of reproduction. We used random amplified polymorphic DNA (RAPD) data to compare the genetic diversity in invasive populations of these species. 42 RAPD bands were identified in B. orientalis and 70 RAPD bands in E. annuus. All six populations of B. orientalis were polymorphic, while only four of eight study populations of E. annuus were polymorphic. The highest differences between species were established at the level of RAPD phenotypes. 67 RAPD phenotypes were found among 90 plants of B. orientalis but only 10 RAPD phenotypes were identified among 89 plants of E. annuus. This study shows that the level of genetic diversity in populations of different invasive species at the same geographic area may vary considerably. Keywords: Bunias orientalis, Erigeron annuus, genetic diversity, RAPD phenotypeVergleich der genetischen Vielfalt von zwei nicht-einheimischen PflanzenartenBunias orientalis und Erigeron annuus sind zwei nicht-einheimische Pflanzenarten, die in litauische natürliche Ökosysteme eindringen. Eindringungs- und Invasionswege beider Arten sind ähnlich. Unterschiede liegen in den Entwicklungs- und Reproduktionsbesonderheiten. In dieser Arbeit haben wir anhand der Ergebnisse von random amplified polymorphic DNA (RAPD) die genetische Vielfalt von verschiedenen Populationen dieser nicht-einheimischen Arten verglichen. 42 RAPD Banden wurden bei B. orientalis und 70 bei E. annuus festgestellt. Alle sechs B. orientalis-Populationen waren polymorph; bei E. annuus waren es nur vier von acht Populationen. Die größte Differenz zwischen den Arten zeigten die RAPD-Phänotypen. Bei 90 B. orientalis-Pflanzen wurden 67 RAPD-Phänotypen identifiziert aber nur 10 RAPD-Phänotypen bei 89 E. annuus-Pflanzen. Die Ergebnisse zeigen, dass die genetische Vielfalt der nicht-einheimischen Pflanzenarten auf demselben geographischen Territorium wesentliche Unterschiede aufweisen kann. Stichwörter: Bunias orientalis, Erigeron annuus, genetische Vielfalt, RAPD-Phänotype

Genotypic diversity and clonal structure of Erigeron annuus (Asteraceae) in Lithuania

This study was conducted to assess the clonal structure and genetic diversity of alien herbaceous plant species Erigeron annuus. The global warming and changes in agriculture practice in the past few decades were favourable for the expansion of this species in Lithuania. We used RAPD and ISSR assays to assess genetic variation within and among 29 populations of E. annuus. A total of 278 molecular markers were revealed. Our study detected reduced level of genetic diversity of invasive populations of E. annuus. Significant differences in DNA polymorphism among populations of E. annuus were also found. Some populations of this species are composed of genetically identical plants, while others were polymorphic. Clonal diversity of study populations ranged from 0.083 to 0.4 for both DNA marker systems. The Simpsons diversity index values ranged from 0.0 to 0.636. The average number of genotypes per population established using both assays was about 1.7. Out of 328 E. annuus individuals only 16 showed unique RAPD and 14 unique ISSR banding patterns. The remaining plants were clones of different size. The most common genotype of E. annuus identified in our study was represented by predominate in nine populations.Keywords: Clonal structure, DNA markers, Erigeron annuus, invasive plants, ISSR, RAPDGenetische Vielfalt und Klonstruktur von Erigeron annuus (Asteraceae) in LitauenZusammenfassungDie Untersuchungen verfolgen das Ziel, die genetische Struktur der nicht-einheimischen Pflanzenart Erigeron annuus auszuwerten. Globale Erwärmung sowie Veränderungen in der landwirtschaftlichen Praxis der letzten Jahrzehnte waren bedeutsam für die Verbreitung dieser Art in Litauen. Für die Feststellung der genetischen Verteilungsvielfalt in den 29 E. annuus Populationen haben wir uns der RAPD und ISSR -Methoden bedient. Insgesamt wurden 117 RAPD - und 161 ISSR - Loci festgestellt. Die Untersuchungen haben die verringerte genetische Vielfalt der E. annuus Populationen aufgezeigt. Außerdem wurden bedeutende DNR polymorphe Unterschiede zwischen E. annuus Populationen angetroffen. Einige Populationen dieser Art bestanden aus genetisch identischen Pflanzen, während die anderen polymorphem waren. Die Klonvielfalt der untersuchten Populationen schwankte zwischen 0.083 und 0.4 bei der Verwendung von beiden DNR-Signifikanten. Die durchschnittliche Genotypen Zahl in der Population betrug etwa 1.7 bei der Verwendung von beiden Signifikanten. Nach der Untersuchung der 328 E. annuus Individuen wurden 16 unikale RAPD - und 14 unikale ISSR - Phänotypen festgestellt. Die übriggebliebenen Pflanzen waren Klonen von unterschiedlicher Größe. Der am meisten verbreitete E. annuus Genotyp wurde in neun Populationen ermittelt.Stichwörter: DNS- Marker, Erigeron annuus, ISSR, Klonstruktur, invasive Pflanzen, RAP

The use of DNA markers in the studies of plant genome and biodiversity

DNA markers are alleles of polymorphic DNA loci that are established using methods of molecular biology and can be used for the identification of specific chromosome region. DNA markers are applied for the detection and analysis of genetic variation. These molecular instruments can help in the understanding of molecular basis of various biological phenomena in plants (loss of genetic diversity, population divergence, adaptivity, response to biotic and abiotic stress, genetic instability et cetera). The use of DNA markers in practical studies requires a careful consideration of the advantages and as well as limitations of various marker techniques. In this review various applications of DNA markers in plant genetic studies including genotyping and characterization of accessions of germplasm collections, assessment of genetic relationships between cultivars, understanding of the genetic variation within and between populations, plant genome analysis and gene cloning are discussed

Investigation of genetic diversity and relationships of blue-berried honeysuckle (Lonicera caerulea L.) cultivars, genetic lines and populations at the Vilnius University Botanical Garden

Blue-berried honeysuckle (Lonicera caerulea L.) as a fruit crop is relatively rare in Lithuania and neighboring countries. Since 1975, the Vilnius University Botanical Garden has collected and investigated blue-berried honeysuckle cultivars. We studied cultivars, genetic lines, and representatives of some populations using molecular marker techniques (RAPD, ISSR, cpDNA). We evaluated the genetic variability and relationships of the studied accessions. Molecular studies indicate that L. caerulea retains high genetic diversity at the intraspecific level

Erigeron annuus genetinės įvairovės gradientas rūšies Europos invazinio arealo dalyje

Erigeron annuus (L.) Pers. (Asteraceae) is native species to eastern North America, but has been introduced to Europe and many other temperate regions of the world. In order to assess the impact of spreading history and settlement time on the genetic diversity of invasive populations of E. annuus, individual plants were sampled from 16 populations located in Switzerland, Poland, Lithuania and Latvia along the south-northern expansion direction of this species in Europe. One population was collected in native range (New Brunswick, Canada). The analysis of ISSR polymorphism in 253 plants revealed 161 polymorphic bands. The highest genotype variation and genetic diversity parameters were revealed in the populations from Switzerland and Poland, the least – in Latvian populations. All 37 plants from Latvia were clones of the same genotype. The comparison of genetic diversity parameters of populations from different countries showed the decrease of genetic diversity on the south-north direction

Chloroplast DNA polymorphism and morphometric characteristics of Carpinus betulus in the Lithuania forests

The European hornbeam (Carpinus betulus L.) is a medium-sized deciduous tree that spreads northeast of the middle of Lithuania. Carpinus betulus L. is a native tree in Poland, and its branch is migrated by two Pleistocene refugia. We hypothesised that its branches had spread to Lithuania. In this study, we selected 10 populations of hornbeam that were chosen from their distribution location. We sequenced the chloroplast intergenic spacer psbA-trnH of 70 individuals. We found 24 bp deletion in chloroplast DNA (cpDNA) individuals of two populations in the southeastern part of Lithuania. In the seven forest populations, we examined the morphological variability of hornbeam seed involucres and nuts variations of 30 morphometric characteristics. Initial genetic population studies were conducted over a wider area; when differences were detected, morphological studies were conducted in the contact zone. Morphometric differences between the study populations were significant. The existence of two haplotypes of cpDNA supports the hypothesis of two migration refugia in C. betulus populations. This study contributes to significant novel knowledge about the morphological and cpDNA variability of European hornbeam populations in Lithuania and Europe

Phylogenetic analysis of the polymorphic 4× species complex Lonicera caerulea (Caprifoliaceae) using RAPD markers and noncoding chloroplast DNA sequences

The blue-berried honeysuckle (Lonicera caerulea L.) is one of the most representative species of the genus Lonicera L. in horticulture. This article presents the results of research on the taxonomy of blue-fruited honeysuckles, which is quite complicated due to the phenotypic plasticity, ability to hybridize and distribution across different ecological zones. We used the random amplified polymorphic DNA (RAPD) markers and sequencing of seven chloroplast DNA (cpDNA) regions (trnH-psbA, rpS12-rpL20, trnL-trnF, trnS-trnG, trnG, rpS16 and trnS-psbZ) to assess the phylogenetic relationships among the taxa within the polymorphic 4× species complex L. caerulea and to determine the position of Lonicera boczkarnikowae Plekh. and Lonicera venulosa Maxim. within this complex. Lonicera chrysantha Turcz. ex Ledeb., L. orientalis Lam. and L. xylosteum L. were used as the outgroup species. The RAPD and cpDNA analyses both indicated that all of the studied taxa of the blue-fruited honeysuckle form a single cluster consisting of two subclusters. A second cluster includes the outgroup species. According to the cpDNA analysis, L. boczkarnikowae and L. venulosa belong to the subcluster that includes the taxa of the polymorphic tetraploid complex L. caerulea. A separate subcluster within the cluster of blue-fruited honeysuckles contains L. altaica and L. edulis

Saxifraga Hirculus L. DNR polimorizmo analizė Lietuvoje

We used RAPDs (random amplified polymorphic DNAs) to analyze genetic diversity in the arctic-alpine yellow marsh Saxifrage (Saxifraga hirculus L.) which is considered to be a glacial relict. Five populations from different regions of Lithuania were studied. A total of 76 individuals were examined in this research. By RAPD analysis 93 loci were detected, of them 71% were polymorphic. All the plants showed different RAPD phenotypes. The percentage of polymorphic bands within populations correlated with population size. UPGMA and PCO analyses showed the genetic specificity of the populations. The genetic distance among individuals in different populations ranged from 0.225 (Juodlė population) to 0.29 (Merkinė population). Some populationspecific minor loci were identified in the Merkinė and Juodlė populations

Assessment of Anthropogenic Impacts on the Genetic Diversity of <i>Phragmites australis</i> in Small-River Habitats

Common reed is often used as a model plant to study the anthropogenic impacts on ecosystems at local and global scales. As a community-forming species, it is directly exposed to the impacts of human activities on the ecosystem. The aim of our study was to evaluate the patterns of genetic diversity in common reed stands located in habitats that are differently affected by anthropogenic factors. We studied whether riverbed modifications, land cover in the neighborhood of the stand and the chemical and physical parameters of the river water affect the genetic diversity of P. australis at the studied sites. Using DNA fingerprinting, we genotyped 747 plants from 42 sites located in 16 small Lithuanian rivers. Bayesian clustering and principal coordinate analysis revealed two main gene pools at the population (river) level. At the site level (i.e., considering all sites independently of their rivers), polymorphism was high even between sites in the same river. Our study revealed a negative relationship between the concentration of nitrogen compounds and the genotypic richness of P. australis populations. We did not find any correlations between the other chemical parameters of the water and the parameters of the genetic diversity. Additionally, there were no genetic differences between sites in modified and unmodified river sections or between sites that differed in land cover type in the neighborhood of the stand

Estimation of genetic diversity and indentity in plus tree clones of Norway spruce (Picea abies (L.) Karst) using RAPD method

The RAPD (random amplified polymorphic DNA) was used to assess genetic diversity and to identify genotype specific markers in Norway spruce (Picea abies (L.) Karst.) plus tree clones. Twenty-three genotypes originally collected from different localities of Lithuania and now growing in a clonal archive at Dubrava experimental-training forest enterprise were analyzed by using 20 oligonucleotide primers. A total of 299 RAPD bands were scored, of which 243 were polymorphic. The estimated genetic distance varied from 0.2905 to 0.5000. These results confirm that studied P. abies clones exhibit a high genetic variation. Fourteen genotype-specific DNA bands were identified. The RAPD technique was also used to check the clonal identity. We compared RAPD profiles of 5 ramets for each of 3 plus tree. Some differences in the RAPD patterns resulting from mis-planting or mis-labeling of cuttings were found. We also established a reference library of RAPD fingerprints for 23 clones, which could serve as a reference source in cases of litigation concerning clone identity. This study confirmed that the RAPD technique is a useful tool for Picea abies genetic analysis and for forestry service in managing propagation operations. Also it could be useful for Norway spruce genetic resources analysis and conservation in Lithuania