Random Amplified Polymorphic DNA Markers Research Articles

Genetic variability in the natural populations of Lasioderma serricorne (F.) (Coleoptera: Anobiidae), detected by RAPD markers and by esterase isozymes.

Lasioderma serricorne (F.) is a small cosmopolitan beetle regarded as a destructive pest of several stored products such as grains, flour, spices, dried fruit and tobacco. Chemical insecticides are one of the measures used against the pest. However, intensive insecticide use has resulted in the appearance of resistant insect populations. Therefore, for the elaboration of more effective control programs, it is necessary to know the biological aspects of L. serricorne. Among these aspects, the genetic variability knowledge is very important and may help in the development of new control methods. The objective of this study was to evaluate the genetic variability of 11 natural populations of L. serricorne collected respectively in three and four towns in the states of Paraná and São Paulo, Brazil, using 20 primers random amplified polymorphic DNA (RAPD) and polymorphisms of esterases. These primers produced 352 polymorphic bands. Electrophoretic analysis of esterases allowed the identification of four polymorphic loci (Est-2, Est-4, Est-5 and Est-6) and 18 alleles. Results show that populations are genetically differentiated and there is a high level of genetic variability within populations. The high degree of genetic differentiation is not directly correlated to geographical distance. Thus, our data indicate that movement of infested commodities may contribute to the dissemination of L. serricorne, facilitating gene flow.

Breeding of Garden Chrysanthemum Cultivar ‘Nuri Ball’ (Dendranthema grandiflorum Ramat.) with White Color Petals and Semi-Decorative Type Characteristics

'누리볼(Nuri Ball)'은 충남농업기술원 예산국화시험장에서 2011년에 육성한 흰색꽃을 가진 반구형 화단국화 품종이다. 품종육성 경위는 모본으로 노랑색 반겹꽃 화형의 줄기가 강한 특 징을 가지고 있는 '02-145-01' 계통과 흰색 반겹꽃 화형의 초형이 안정적인 '02-04-32'계통을 2004년에 인공교배하여 얻었으며, 2006년부터 2009년까지 특성을 조사하였고, 농작물 직무육성 신품종 선정위원회를 거쳐 2011년에 품종등록을 하였다. '누리볼'의 화색은 흰색이고 반겹꽃 화형의 형태를 가졌다. 생육에서 초장은 대조품종인 '화이트 미리' 보다 작으나, 초폭이 더 넓은 안정적인 반구형의 모습을 보여 화단국화로써 뛰어난 형태가 보이는 것으로 판단된다. 대조 품종과 형태적으로는 다소 유사 하였지만, 유전적 관계와 배수성 검정을 통하여 품종의 차별성을 확인하였다. '누리볼'은 5월 하순에 정식하면 9월 하순에 개화되어 대조 품종보다 16일 빠른 개화특성을 보였으며, 꽃은 대조품종인 '화이트 미리' 보다 크지만 꽃수는 적은 것으로 나타났다. '누리볼'은 화단에 이용 가능한 조경용 국화로써, 앞으로 농가소득 창출에 기여할 수 있을 것으로 기대된다. The new garden chrysanthemum (Dendranthema grandiflorum Ramat.) cultivar 'Nuri Ball' was developed at Yesan Chrysanthemum Experiment Station of Chungcheongnam-do Agricultural Research and Extension Services in 2011. 'Nuri Ball' was bred through a cross between the '02-145-01' line as the female parent with yellow flower color and '02-04-32' as the male plant with white flower color in 2004. Three years of adaptation trials were conducted from 2006 to 2009 under natural conditions. This study compared the external shape type with that of 'White Miri' and conducted ploidy and RAPD (Random amplified polymorphic DNA) marker analyses. These tests showed that 'Nuri Ball' cultivar has its own characteristics compared with the control 'White Miri'. 'Nuri Ball' was a shrub type variety with semi-double flowers of 4.0 cm in width with white petals. It could produce 1025.2 flowers per plant in autumn. Compared with the control 'White Miri', 'Nuri Ball' was similar in terms of shape and color of flowers, but was different in flower size and number. The natural flowering time of 'Nuri Ball' was late September. It had very vigorous growth and an early budding plant. 'Nuri Ball' was demonstrated to be a new cultivar based on ploidy test and RAPD analysis. 'Nuri Ball' is intended for use as a bed chrysanthemum and expected to contribute to farm incomes in landscaping.

Genetic diversity of the bacterial wilt pathogen Ralstonia solanacearum using a RAPD marker

Bacterial wilt caused by Ralstonia solanacearum is a destructive disease of many economically important crop species. A significant variation in wilt incidence and severity in eggplant and potato was observed among the growing areas surveyed. R. solanacearum isolates obtained both from eggplant and potato belong to biovar III, while isolates from eggplant belong to race 1 and isolates obtained from potato belong to race 3. Random amplified polymorphic DNA (RAPD) technique was used as a tool for assessing genetic variation and relationship among seven isolate groups of R. solanacearum viz., RsB-1, RsB-2, RsB-3, RsP-1, RsP-2, RsP-3 and RsP-4, consisting in a total of 28 isolates. Out of the RAPD markers used, amplification with four decamer primers produced 70 bands with sizes ranging from 100 to 1400bp. Out of 70 bands, 68 bands (97.06%) were polymorphic and two bands (2.94%) were monomorphic amongst the seven R. solanacearum isolates group. The Unweighted Pair Group Method of Arithmetic Means (UPGMA) dendrogram constructed from Nei's genetic distance produced two main clusters of the seven isolates of R. solanacearum. The isolates RsB-1, RsB-2, RsB-3 and R-4 grouped in cluster І, while RsP-2, RsP-3 and RsP-4 grouped in cluster ІІ. The highest intra-variety similarity index (Si) was found in RsB-1 isolate (86.35%) and the lowest one in RsP-2 (56.59%). The results indicated that relatively higher and lower levels of genetic variation were found in RsP-3 and RsB-3, respectively. The coefficient of gene differentiation (Gst) was 0.5487, reflecting the existence of a high level of genetic variations among seven isolates of R. solanacearum. Comparatively higher genetic distance (0.4293) and lower genetic identity (0.6510) were observed between RsB-2 and RsP-4 combinations. The lowest genetic distance (0.0357) and highest genetic identity (0.9650) were found in RsB-1 vs. RsB-2 pair. Thus, RAPD offers a potentially simple, rapid and reliable method to evaluate genetic diversity analysis in R. solanacearum.

Evaluation of genetic diversity on Prunus mira Koehne by using ISSR and RAPD markers

Peach (Prunus mira L.; 2n = 16), which originated in China, is a fruit tree widely cultivated all over the world with high economic value. Prunus mira Koehne is a wild species distributed in Tibet with special medicinal and ecological values. To conserve its breeding and germ plasm resources, two DNA-based molecular markers, inter-simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD), were used for genetic diversity evaluations of 48 samples from three regions, including Tibet (XZ), Henan Province (HN) and Heilongjiang Province (HLJ), in this study. High levels of polymorphism were observed: 77.80% (ISSR) and 72.73% (RAPD), which indicated that ISSR and RAPD were efficient methods to detect the genetic diversity of P. mira Koehne. The samples from Tibet preserved higher genetic diversity and most genetic variations occurred among populations. There was no significant relationship observed between the genetic distance of populations and their geographic distribution. For conservation of P. mira Koehne, ex situ conservation is a suitable strategy that needs to be established in the future.

Genetic diversity and phylogenetic relationships between and within wild Pistacia species populations and implications for its conservation

Although cultivation and utilization of Pistacia are fully exploited, the evolutionary history of the Pistacia genus and the relationships among the species and accessions is still not well understood. The aim of this study was to analyze random amplified polymorphic DNA (RAPD) in a total of 50 accessions of wild pistachio species, which included five populations Pistacia vera, Pistacia khinjuk, Pistacia atlantica, Pistacia mutica, and Pistacia eurycarpa. High levels of genetic diversity were detected within wild pistachio accessions, as revealed by using the unweighted pair-group method with arithmetic averaging and supported via analysis of molecular variance. The objectives of this investigation were to estimate marker indices, polymorphic information contents (PICs), and genetic similarities (GS) for RAPD markers; assess the genetic diversity of Pistacia species, using GS estimated from RAPD fingerprints and molecular characterization; and facilitate the use of markers in inter-specific introgression and cultivar improvement. Out of the 149 polymerase chain reaction fragments that were scored, 146 (97.98 %) were polymorphic. Genetic similarities ranged from 0.3 to 0.86 %, marker indices ranged from 2.98 to 17.74 %, and PICs ranged from 0.80 to 0.99 %. Our results provided great molecular identification of all assayed genotypes, which have shown that there is large quantity of genetic diversity among the pistachio accessions. This finding might render striking information in breeding management strategies for genetic conservation and cultivar development.

Determination of Genetic Diversity Among Wild Grown Apples From Eastern Black Sea Region in Turkey Using ISSR and RAPDs Markers

In this study, we investigated interspecific variations of apple genotypes using ISSR (Inter-Simple Sequence Repeats) and RAPD (Randomly amplified Polymorphic DNA) technique. Total 19 genotypes of wild grown local varieties were collected from Eastern Black Sea region of Turkey and have been studied for the analysis of ISSR and RAPD profile to examine their relationship. From total 53 RAPD and ISSR primer investigated, 28 could amplify clearly and consistently. All bands obtained from ISSR and RAPDs primer were polymorphic.

Identification of ISSR and RAPD markers linked to yield traits in bread wheat under normal and drought conditions.

Genetic variability and identification of some molecular markers were studied in twenty promising lines of wheat using agronomic traits, ISSR (inter simple sequences repeats) and RAPD (random amplified polymorphic DNA) markers. Significant variation was evidenced in all agronomic traits. The lines proved to be superior to the check cultivar Sahel1 in yield and its component traits. Lines L2, L7 and L8 were the best in most yield component traits in both seasons. Moreover, Lines L2, L4, L5, L7 and L8 showed drought tolerance by which they displayed high performance in agronomic traits as well as a low drought susceptibility index. The percentage of polymorphism was 39.3% and 53.2% for ISSRs and RAPDs, respectively. UBC-881 belonged to penta-nucleotide repeat sequences (GGGTG) that produced the highest level of polymorphism, while UBC-846 belonged to di-nucleotide repeat sequences (CA) that produced the lowest level of polymorphism. Genetic similarities among wheat lines based on ISSR and RAPD markers ranged from 0.81 to 1.00 and from 0.86 to 0.98, respectively. There was a low average of PIC (polymorphism information content) values which were 0.10 (ISSR) and 0.15 (RAPD). The RAPD technique exhibited a higher marker index (MI = 0.69) compared to ISSR (MI = 0.43). There was insignificant correlation between ISSR and RAPD data (0.168, p > 0.05). There were two markers (UBC-881450bp and OPF-10540bp), on each of which two traits regressed significantly. The associated markers each explained a maximum regression of 18.92–34.95% of the total available variation for individual associated traits.

Assessment of genetic variations of silver lime (Tilia tomentosa Moench.) by RAPD markers in urban and forest ecosystems

In the present study, the genetic diversity analysis of Tilia tomentosa plants was performed by using random amplified polymorphic DNA (RAPD) primers. Twenty eight plant samples, collected from urban (25 members) and forest (3 members) ecosystems, were used in this study. A total of 53 bands were obtained from eight RAPD primers, of which 48 (90.6%) were polymorphic. The percentage of polymorphic loci (P) was found to be 94.29%, the observed number of alleles (Na) was 1.94, the effective number of alleles (Ne) was 1.60, Nei's gene diversity (h) was 0.34 and Shannon's information index (I) was 0.50. Unweighted pair group method with arithmetic mean (UPGMA) cluster analysis revealed two major groups. Members of the urban and forest ecosystems showed high genetic similarity (28%–92%) and they did not separate from each other in UPGMA tree. Furthermore, urban and forest genotypes clustered together in principal component analysis.

English

While random amplification of polymorphic DNA (RAPD) markers linked to disease resistance genes have been widely used in plant breeding programs, they generally lack reproducibility. To overcome this major disadvantage and other drawbacks, RAPD markers can be converted into sequence characterized amplified region (SCAR) markers, which are genetically defined loci amplified by polymerase chain reaction (PCR) using specific primers. Thus, SCAR markers are typically more reproducible than RAPD markers, due to specific amplification of genomic regions. In this study, a previously identified RAPD marker AT9/917 that is linked to the Puccinia psidii Winter (rust) resistance gene 1 (Ppr1) in Eucalyptus grandis was successfully converted into a specific SCAR marker. Seven specific SCAR primers were designed based on cloning and sequencing of the RAPD marker AT9/917. Different pairs of SCAR primers were tested in an E. grandis family from a crossing between a resistant and a susceptible E. grandis. Prime pair SCAR AT99151L and AT9915914R produced amplicons of expected size. Restriction enzyme digestion of the amplicon revealed polymorphisms between the resistant and susceptible parents. Association analysis between phenotype (rust resistance) and SCAR genotypes in the E. grandis family suggests that this specific SCAR is useful for marker-assisted selection of E. grandis trees resistance to P. psidii Winter. Key words: Plant breeding, molecular markers, random amplified polymorphic DNA (RAPD), Mark-assisted selection, sequence characterized amplified region (SCAR).

Genetic diversity in Cucurbita pepo landraces revealed by RAPD and SSR markers

Cucurbita pepo landraces are important traditional leafy vegetable crops that are widely eaten in KwaZulu-Natal, South Africa, and elsewhere. The use of molecular markers is beneficial in the genetic study of landraces with different agro-ecological origins. In this study, genetic variation in seven selfed and unselfed C. pepo landraces from three districts in the KwaZulu-Natal Province was investigated using the random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers. Of the 36 and 55 primers tested, 9 RAPD and 10 SSR primers were selected for their reproducibility and high polymorphism. A total of 100 fragments were detected by RAPD, out of which 94 (94%) fragments were considered polymorphic. SSR markers revealed a total of 56 alleles, where 38 (68%) alleles were polymorphic. The sizes of fragments ranged from 75 to 1800bp and from 124 to 251bp in RAPD and SSR markers, respectively. The number of fragments per primer ranged between 9 and 14 and between 1 and 12, in RAPD and SSR, respectively. The genetic differentiation coefficient between populations (GST) ranged between 0.0022 and 0.0100 with RAPD marker and between zero and 0.0076 with SSR marker. Both markers revealed overwhelming averages of gene flow: 97.78 and 132.09 in RAPD and SSR markers, respectively. Both techniques discriminated the landraces very effectively, but only the RAPD marker was able to discriminate landraces according to fruit colour change at maturity as well as their agro-ecological origins. Dendrograms of both markers depicted the close relationship between landraces originating from the Umkhanyakude district (MNS and MS landraces). All genetic parameters indicated that there was plentiful genetic diversity in C. pepo landraces of northern KwaZulu-Natal, South Africa.

Efficacy of RAPD, ISSR and DAMD markers in assessment of genetic variability and population structure of wild Musa acuminata colla.

North east India is considered as one of the major biodiversity hotspots worldwide and centre of origin of several plant species including Musa. Musa acuminata Colla is known to be one of the wild progenitors of cultivated bananas and plantains. Three single primer based DNA marker techniques viz., random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) and directed amplification of minisatellites DNA (DAMD) were used for diversity diagnostics among 25 genotypes of wild M. acuminata collected from Meghalaya province of north east India. A total of 58 primers (26-RAPD, 21-ISSR, and11-DAMD) yielded 451 DNA fragments, of which 395 (87.58%) were found to be polymorphic in nature. The polymorphic information content (PIC) values were almost identical for each marker system. The resolving power of the marker system was found to be highest in RAPD (3.96) whereas ISSR resolved highest marker index (16.39) in the study. Selected amplicon data obtained through single primer amplification reactions were utilized for determination of diversity within and among the populations of M. acuminata. Nei's genetic differentiation (Gst) value (0.451) indicated higher proportion of the genetic variation within the populations which is supported by the AMOVA analysis (88%). The study provides insight into the efficacy of RAPD, ISSR and DAMD to analyse the genetic variation existing in the wild Musa germplasm, which can furtherbe exploited for quality trait improvement and domestication of such important horticultural crops. The genetic diversity based population structure may shed light on the genetic basis of speciation and evolution of various species within the genus Musa.

Divergent selection along climatic gradients in a rare central European endemic species, Saxifraga sponhemica.

Background and Aims The effects of habitat fragmentation on quantitative genetic variation in plant populations are still poorly known. Saxifraga sponhemica is a rare endemic of Central Europe with a disjunct distribution, and a stable and specialized habitat of treeless screes and cliffs. This study therefore used S. sponhemica as a model species to compare quantitative and molecular variation in order to explore (1) the relative importance of drift and selection in shaping the distribution of quantitative genetic variation along climatic gradients; (2) the relationship between plant fitness, quantitative genetic variation, molecular genetic variation and population size; and (3) the relationship between the differentiation of a trait among populations and its evolvability.Methods Genetic variation within and among 22 populations from the whole distribution area of S. sponhemica was studied using RAPD (random amplified polymorphic DNA) markers, and climatic variables were obtained for each site. Seeds were collected from each population and germinated, and seedlings were transplanted into a common garden for determination of variation in plant traits.Key Results In contrast to previous results from rare plant species, strong evidence was found for divergent selection. Most population trait means of S. sponhemica were significantly related to climate gradients, indicating adaptation. Quantitative genetic differentiation increased with geographical distance, even when neutral molecular divergence was controlled for, and QST exceeded FST for some traits. The evolvability of traits was negatively correlated with the degree of differentiation among populations (QST), i.e. traits under strong selection showed little genetic variation within populations. The evolutionary potential of a population was not related to its size, the performance of the population or its neutral genetic diversity. However, performance in the common garden was lower for plants from populations with reduced molecular genetic variation, suggesting inbreeding depression due to genetic erosion.Conclusions The findings suggest that studies of molecular and quantitative genetic variation may provide complementary insights important for the conservation of rare species. The strong differentiation of quantitative traits among populations shows that selection can be an important force for structuring variation in evolutionarily important traits even for rare endemic species restricted to very specific habitats.

English

This study presents the results from comparison of random amplified polymorphic DNA (RAPD), random amplified polymorphic DNA (RAPD) and RAPD-ISSR markers together in determining the genetic diversity and polymorphism among strains of fluorescent Pseudomonas. In order to compare the efficiencies of these markers, 17 different isolates were investigated using RAPD and ISSR. In RAPD marker analysis, 147 out of 222 bands (63.85%) were polymorphic and the RAPD based genetic similarity (RAPD-GS) ranged from 0.11 to 0.73. In ISSR analysis, a total of 134 alleles were detected, among which 105 alleles (77.9%) were polymorphic. The ISSR derived genetic similarity (ISSR-GS) ranged from 0.38 to 0.81. Cluster analysis indicated that all the 17 isolates could be distinguished by both RAPD and ISSR markers and worked effectively to determine high level of polymorphism in Pseudomonas. But ISSR was found to be slightly better than RAPD in assessing genetic diversity among the isolates while the combination of these techniques will give the comprehensive genetic analysis of Pseudomonas. Principal component analysis (PCA) was employed to evaluate the resolving power of the markers to differentiate between them. Key words: Fluorescent Pseudomonas, Molecular markers, RAPD , ISSR, PCR, Genetic diversity, Phylogeny, Principal component analysis.

Molecular Characterization of Some Turkish Olive Cultivars Using Random Amplified Polymorphic DNA (RAPD) Markers

Olive (Olea europea L.) is one of the oldest cultivated plants characteristic in the Mediterranean area, where it is the most important oilproducing crop. The cultivated olive (O. europaea L. var. europaea) is propagated by cutting or grafting, whereas wild olive (O. europaea L. var. sylvestris) is reproduced from seeds. These two olive types are interfertile and have led to a large number of varieties. Morphological descriptions are not entirely reliable, due to numerous synonyms and homonyms in designations, labelling mistakes, the presence of varietal clones, and the uncertain identification methods thus far applied. Molecular markers, as random amplified polymorphic DNA (RAPD) markers, are environment-independent and efficient to identify olive varieties and to detect synonymous and homonymous. In this study, fifteen selected RAPD markers are used for determination of relationships among twenty individuals belonging to four important Turkish olive cultivars. Our results showed that RAPD markers can be used to differentiate olive cultivars

Genetic characterization of microsporidians infecting Indian non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) by using PCR based ISSR and RAPD markers assay

This study established the genetic characterisation of 10 microsporidian isolates infecting non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) collected from biogeographical forest locations in the State of Assam, India, using PCR-based markers assays: inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD). A Nosema type species (NIK-1s_mys) was used as control for comparison. The shape of mature microsporidian spores were observed oval to elongated, measuring 3.80 to <TEX>$4.90{\mu}m$</TEX> in length and 2.60 to <TEX>$3.05{\mu}m$</TEX> in width. Fourteen ISSR primers generated reproducible profiles and yielded 178 fragments, of which 175 were polymorphic (98%), while 16 RAPD primers generated reproducible profiles with 198 amplified fragments displaying 95% of polymorphism. Estimation of genetic distance coefficients based on dice coefficients method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was done to unravel the genetic diversity of microsporidians infecting Indian muga and eri silkworm. The similarity coefficients varied from 0.385 to 0.941 in ISSR and 0.083 to 0.938 in RAPD data. UPGMA analysis generated dendrograms with two microsporidian groups, which appear to be different from each other. Based on Euclidean distance matrix method, 2-dimensional distribution also revealed considerable variability among different identified microsporidians. Clustering of these microsporidian isolates was in accordance with their host and biogeographic origin. Both techniques represent a useful and efficient tool for taxonomical grouping as well as for phylogenetic classification of different microsporidians in general and genotyping of these pathogens in particular.

Analysis of Genetic Diversity in 21 Genotypes of Indian Banana Using RAPDs and IRAPs Markers

Table top banana (AAA) and cooking type plantines (BBB/ABB) are cultivated commercially for high yield and lucrative market value. Morphological similarities among the banana clones make identification difficult under field conditions. Random amplified polymorphic DNA (RAPD) and inter-retrotransposon amplified polymorphism (IRAP), were used to characterize genetic variations among 21 banana germplasm. IRAP primers were designed to determine ‘AA’ and ‘BB’ specific markers on the basis of repetitive and genome-wide dispersed long terminal repeat (LTR) retrotransposons. RAPD markers successfully detected genetic variation between genotypes. IRAP markers amplified either by a single primer or a combination of primers, based on LTR orientation, successfully amplified different retrotransposons dispersed in the Musa genome. The average level of polymorphism exhibited by RAPD and IRAP markers were 71.47 % and 81.3 % respectively that suggests substantial genetic variations among the tested varieties. All the 12 table-top varieties were clustered together while four cooking varieties i.e. Bantala-I, Bantala-II, Dakhkhnisagar and Athiakol with ‘BB’ formed a distinct group. These results suggested that IRAP markers were found more robust than RAPD markers to study the intra-group genetic diversity between table top and cooking banana.

Female-specific RAPD and SCAR Markers in Celastrus orbiculatus

Celastrus orbiculatus Thunb. belongs to Celastrus L. of the family Celastraceae. As a kind of traditional Chinese herbal medicine,it has great ornamental value and important medicinal value. It is a dioecious vine,a small amount of them are andromonoecy plants. However,the morphological differences between the male and female plants are not obvious before the maturation of their sex organs,which restricts the early cultivation and breeding. Currently,molecular markers have been widely used in the sex identification of dioecious plants. But the research on the sex differentiation of C. orbiculatus at the molecular level has not been reported. Due to unknown genome sequence,many studies on C. orbiculatus are limited in the field of chemistry and cultivation technology,which greatly limits its utilization. In this study,differentiation among dioecious plants was compared using random amplified polymorphic DNA(RAPD) markers. It is the first time to use the RAPD markers on the study of C. orbiculatus. Using the young leaves of male and female plants as materials,we extracted the DNA using the improved CTAB method. Then through screening of different factors,we established the optimized PCR system. Based on the above PCR system,we selected 100 RAPD primers to conduct the analysis. Significantly different bands were amplified using primer S127, S140, S148, S174 and S111,respectively. These different bands were cloned into the T-vector,and then sequenced. Based on the sequences,the specific SCAR markers were obtained. Only one female-associated dominant SCAR marker from RAPD marker S111 was verified. Sequence analysis showed that this female specific fragment contained nine open reading frames(ORF) between 93- 570 bp in length,among which two have more than 100 amino acids. The results of sequence Blast showed that the polypeptide fragment had a higher similarity with plants reverse transcription factor of the gag-POL protein,which belongs to UBN2 super family. We inferred a possible reason of the gender traits is that the insertion of the reversal seat factor may inhibit some important sex-determining factors. In the future,we need to get their full-length sequences using RACE,and then study their function on sex differentiation.

Evaluating the Genetic Relatedness within Lupinus Pilosus L. Species Based on RAPD Analysis

Nine Random Amplified Polymorphic DNA (RAPD) primers showing polymorphic bands were used to determine the genetic relatedness among and within single individual plants of Lupinus pilosus and used for the construction of the dendrogram and the similarity matrix. A total of 1112 bands were obtained, 219 of which were polymorphic. Similarity values among the studied single individuals of Lupinus pilosus ranged from 0.450 to 0.02. The cluster analysis obtained by Unweighted Pair-Group Method with arithmetic averages (UPGMA) grouped the tested individuals into three clusters. A genetic variation was found within Lupinus pilosus based on RAPD markers. The variability emphasis the presence of genetic diversity within Lupinus pilosus species.

Genetic Variation of &lt;I&gt;Salmo trutta&lt;/I&gt; L. Populations from the Catchment Areas of the Rega, Parseta and Wieprza Rivers Evaluated by RAPD and SSR Markers

By using simple sequence repeats (SSR) and random amplification of polymorphic DNA (RAPD) markers, the genetic variability of three Salmo trutta L. populations from three rivers, the Wieprza, the Rega and the Parseta, was determined. The investigated populations showed a high level of genetic variability. Microsatellites showed that observed heterozygosity (Ho) was higher than the expected heterozygosity (He), with most heterozygotes found in the population from the Parseta river and the fewest in the Wieprza population. The F(IS) coefficient in all investigated populations of the sea trout indicate a high excess of heterozygotes. The highest genetic differentiation was observed between the sea trout from the Rega river and those from the Wieprza (0.366). The obtained results based on microsatellite and RAPD analysis showed that the investigated populations formed two groups. The first group consisted of the sea trout populations from the Wieprza and the Parseta rives, while the second group was formed solely by the Rega river population.

Assessment of genetic characteristics of Aconitum germplasms in Xinjiang Province (China) by RAPD and ISSR markers

Aconitum is a medicinal treasure trove that grows extensively on fertile pastures in Xinjiang Province (China); however, its molecular genetic characteristics are still poorly studied. We studied Aconitum kusnezoffii Reichb., Aconitum soongaricum Stapf., Aconitum carmichaelii Debx. and Aconitum leucostomum Worosch, using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) techniques, to evaluate their genetic relationship and potential medicinal value. Our results showed that A.kusnezoffii Reichb. and A.soongaricum Stapf. have close genetic relationship and cluster together. Polymorphism rates of 97.25% and 98.92% were achieved by using 15 RAPD and 15 ISSR primers, respectively. Based on Nei's gene diversity (H) and Shannon's index (I), the inter-population diversity (Hs) was higher when compared with the intra-population diversity (Hp). Among the three Aconitum populations, the coefficient of gene differentiation (Gst) was 0.4358 when evaluated by RAPD and 0.5005 by ISSR. The genetic differentiation among the three Aconitum populations was highly significant, suggesting low gene flow (Nm). This was confirmed by the estimates of gene flow (Nm = 0.6473 and Nm = 0.4991, based on ISSR and RAPD data, respectively). Comparing the RAPD and ISSR results, the two DNA markers proved similarly effective in the assessment of the genetic characteristics of the studied Aconitum populations and could be used for reliable fingerprinting and mapping in studies on Aconitum diversity in view of Aconitum suitability for development and protection.