Search

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.

Genus Batrachium (DC.) Gray is evolutionary young but very important group of aquatic plants. It is very essential component in the river for its ability to affect flow dynamics, silt deposition and provide refuge for ecologically important macroinvertebrate and fish populations and can be considered as the indicator of the sustainability of riverine ecosystem. Communities with Batrachium species are regarded as significant for European Union and listed in Apendix 1 of Habitat Directive (code 3260). Plants of this genus are important not only from the scientific point, but also and in terms of plant conservation. By reason of this, studies of morphological features of Batrachium species are very relevant. Our study is an introduction to further detail research of Batrachium species. It was performed in 16 river catchments. During this studies 185 Batrachium individuals were collected and measured. The morphological features such as the presence of floating leaves, number of capillary leaves furcations, length of terminal leaf segments, length of petiole, hairiness of stipules, length and number of petals, and hairiness of receptacle, were used in this analysis. The species level taxonomic identification of the individuals was performed using keys after Cook (1966), Webster (1988), Dahlgren, Jonsell (2001), Bobrov (2003), Webster, Rich (1998). Nonparametric Kruskal-Wallis test was used to compare morphological features. Mann-Whitney U test was used to identify differences between specific populations. The analysis showed that not all analysed features were uniformly important in Batrachium species identification. Acording to some morfological features all samples were grouped in two groups. Individuals from the first group distinguished glabrous or sparse hairy receptacle, had 5-8 petals (length ranged from 0.4 to 1.1 cm), had only capillary leaves, length of secondary leave ranged from 4 to 9 cm, and length of the sixth leave ranged from 6 to 12 cm. While specimens from another group had densely pubescent receptacle, five petals (length ranged from 0.2 to 0.5 cm), length of secondary capillary leaves ranged from 3 to 7 cm and length of the sixth leave ranged from 5 to 8 cm. Some individuals from Skroblus, Ūla and Merkys populations had intermediate leaves. We suppose, that individuals from first group can be described as Batrachium fluitans, whereas plants from second group – as Batrachium penicillatum s.l. or Batrachium trichophyllum.

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.

Genetic diversity of Primula farinosa L., an endangered species in Lithuania, was analyzed using RAPD method. Four populations from different places of Lithuania were studied. Totally, 55 RAPD bands were revealed from 7 primers. 29,1 % of RAPD bands were polymorphic. PCoA showed genetic specificity of studied populations.

DNR žymenys yra polimorfinių DNR lokusų aleliai, lokalizuoti tam tikrose genomo vietose ir nustatomi naudojant įvairius molekulinės biologijos metodus. Genetinės įvairovės nustatymas ir tyrimas naudojant DNR žymenis gali padėti suprasti daugelio biologinių reiškinių, vykstančių augaluose (ląstelės, organizmo, rūšies lygmenyje), prigimtį. Vykdydami aptariamus tyrimus norime rasti atsakymą į pagrindinį klausimą: kokios naudotų DNR žymenų galimybės kai kurioms augalų biologijos problemoms (genetinių išteklių išsaugojimo, adaptyvumo, populiacijų diferenciacijos, biotinio ir abiotinio streso ir kt.) spręsti? Naudodami įvairius DNR žymenis kaip molekulinius instrumentus, įvertinome genetinės įvairovės lygį pagrindinėse miežių veislėse, sukurtose Baltijos šalyse ir Baltarusijoje, pritaikėme DNR žymenų nustatymo metodus Rubus idaeus, Lonicera ceruleae genetinėse kolekcijose esančių pavyzdžių genotipavimui bei miško medžių (Pinus sylvestris, Piceae abies) rinktinių medžių klonų tapatumo nustatymui; naudodami EST žymenis klonavome ir ištyrėme du genus, dalyvaujančius Solanum tuberosum atsake į biotinį (Erwinia carotovora infekcija) ir abiotinį stresą. Naudodami RAPD metodą ištyrėme Saxifraga hirculus, Piceae abies, Fraxinus excelsior, Taxus baccata, Rubus idaeus, Pinus sylvestris populiacijų genetinę struktūrą, diferenciacijos lygį, kai kurių ekologinių veiksnių įtaką genetinės įvairovės pasiskirstymui populiacijose. 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.

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. RAPD metodu buvo tirta pelkinės uolaskėlės (Saxifraga hirculus L.) genetinė įvairovė. Ištirtos penkios populiacijos iš skirtingų Lietuvos rajonų. Šiuo metodu išaiškinti 93 lokusai, iš kurių 71% buvo polimorfiškas. Nustatytas ryšys tarp tirtų individų skaičiaus ir DNR polimorfizmo populiacijoje. Didžiausias DNR polimorfizmas rastas Merkinės (86,4%), o mažiausias – Juodlės populiacijoje (71,2%). UPGMA ir PCO metodai išryškino tirtų populiacijų genetinį savitumą. Genetinis atstumas tarp individų populiacijose svyravo nuo 0,225 (Juodlės populiacija) iki 0,29 (Merkinės populiacija). Merkinės ir Juodlės populiacijose buvo nustatyta keletas populiacijoms specifinių lokusų.

In 2003–2006 researchers of the Pomology Department of Vilnius University Botanical Garden studied 34 blue-berried honeysuckle lines and 9 cultivar-standards. Half of investigated lines (17 of 34) have berries of more than 0.70 g weight: ‘2E’, ‘2S’, ‘32’, ‘22T12’, ‘1B43’, ‘1E’, ‘96-3’, ‘2-1’, ‘96-4’, ‘639-8’, ‘1L’, ‘2R’, ‘2C’, ‘3–5’, ‘Kairėnai’, ‘L-69-3’, ‘3U’. The dry matter of berries varied from 12.21 (‘2C’) to 21.03 (‘1P’), concentration of sugar – from 5.02 (‘1L’) to 9.22 (L. caerulea), acidity – from 1.03 (‘Baktcharskaja’) to 2.45 (‘2C’). There were 171 samples from infected plants collected and 9 fungal disease agents were identified. Plants of L. caerulea, L. caerulea subsp. kamtschatika, 14 lines (‘1L’, ‘1M’, ‘1P’, ‘10’, ‘32’, ‘2C’, ‘2K’, ‘2R’, ‘2S’, ‘3-5’, ‘3R’, ‘3U’, ‘96-4’, ‘L69-3’) and 7 cultivars (‘Goluboje vereteno’, ‘Morena’, ‘Narymskaja’, ‘Tomichka’, ‘Vasjuganskaja’, ‘Viola’, ‘Zoluschka’) were resistant to the impact of fungal disease agents. There are 6 lines, which include both (large berries and resistance to disease) features: ‘1L’, ‘2C’, ‘3U’, ‘3-5’, ‘32’, ‘96-4’. Genetic variation and relationships among 9 blue-berries honeysuckle lines and cultivars based on RAPD analysis was determined. Cultivar ‘Morena’ and line ‘L-69-3’ are genetically distinct from the other investigated lines and cultivars. 2003-2006 metais VU Botanikos sodo melsvauogių sausmedžių kolekcijoje tirta 34 linijos ir 9 veislės-standartai. Iš 34 tirtų linijų daugiau negu 0,70 g sveriančias uogas turėjo pusė – 17 linijų (‘2E’, ‘2S’, ‘32’, ‘22T12’, ‘1B43’, ‘1E’, ‘96-3’, ‘2-1’, ‘96-4’, ‘639-8’, ‘1L’, ‘2R’, ‘2C’, ‘3-5’, ‘Kairėnai’, ‘L69-3’, ‘3U’). Sausų medžiagų kiekis uogose svyravo nuo 12,21 (‘2C’) iki 21,03  (‘1P’), cukraus – nuo 5,02 (‘1L’) iki 9,22 (L. caerulea), rūgštingumas – nuo 1,03 (‘Baktcharskaja’) iki 2,45  (‘2C’). Kolekcijoje buvo surinktas 171 ligų pažeistas augalo antžeminės dalies pavyzdys, identifikuoti 9 grybinių ligų sukėlėjai. Nustatytos atspariausios linijos ir veislės: L. caerulea, L. caerulea subsp. kamtschatika, 14 linijų (‘1L’, ‘1M’, ‘1P’, ‘10’, ‘32’, ‘2C’, ‘2K’, ‘2R’, ‘2S’, ‘3-5’, ‘3R’, ‘3U’, ‘96-4’, ‘L69-3’) ir 7 veislės (‘Goluboje vereteno’, ‘Morena’, ‘Narymskaja’, ‘Tomichka’, ‘Vasjuganskaja’, ‘Viola’, ‘Zoluschka’). Šešiose linijose (‘1L’, ‘2C’, ‘3U’, ‘3-5’, ‘32’, ‘96-4’) dera abu požymiai: uogų stambumas ir atsparumas grybinėms ligoms. Kai kurių pavyzdžių genotipavimas RAPD metodu parodė, kad visos devynios tirtos linijos ir veislės yra savitos, bet labiausiai nuo daugumos skiriasi veislės ‘Morena’ ir linijos ‘L69-3’ augalai.

APPD (atsitiktinai pagausinta polimorfinė DNR; angl. RAPD, random amplified polymorphic DNA) metodas buvo panaudotas paprastosios eglės (Picea abies (L.) Karst.) pliusinių medžių klonų genetinei įvairovei įvertinti bei genotipo specifiniams žymenims nustatyti. Tirti 23 paprastosios eglės klonai, kilę iš įvairių Lietuvos vietovių ir auginami kloniniame archyve, esančiame Dubravos eksperimentinėje-mokomojoje miškų urėdijoje. Tyrimams buvo panaudota 20 oligonukleotidinių pradmenų. Iš viso nustatyti 299 atsitiktinai pagausinti DNR fragmentai, iš kurių 243 – polimorfiniai. Rezultatai atskleidė didelę tirtų klonų genetinę įvairovę. Genetinis atstumas svyravo nuo 0,2905 iki 0,5000. Nustatyta 14 genotipui specifinių molekulinių žymenų. APPD metodas taip pat buvo panaudotas klonų tapatumui patikrinti. Šiam tyrimui panaudoti trijų paprastosios eglės pliusinių medžių klonai (kiekvieno po 5 rametas). Palyginus APPD spektrus, buvo nustatyta keletas netikslumų, greičiausiai atsiradusių dėl klonų sodinimo ar žymėjimo klaidų. Taip pat buvo sukurta 23 klonų APPD spektrų biblioteka, kuri gali būti naudojama klonų tapatumui įvertinti. Tyrimai patvirtino, kad APPD metodas yra efektyvus ir patogus paprastosios eglės genetinei įvairovei tirti bei dauginamosios medžiagos tapatumo kontrolei miškų ūkyje atlikti. Šis metodas gali būti naudojamas Lietuvos paprastosios eglės genetiniams ištekliams analizuoti, tvarkyti bei išsaugoti. 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.

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.