Rosaceous Species Research Articles

Phosphorylation of aldose-6-phosphate reductase from Prunus persica leaves

Sugar-alcohols are major photosynthates in plants from the Rosaceae family. Expression of the gene encoding aldose-6-phosphate reductase (Ald6PRase), the critical enzyme for glucitol synthesis in rosaceous species, is regulated by physiological and environmental cues. Additionally, Ald6PRase is inhibited by small molecules (hexose-phosphates and inorganic orthophosphate) and oxidizing compounds. This work demonstrates that Ald6PRase from peach leaves is phosphorylated in planta at the N-terminus. We also show in vitro phosphorylation of recombinant Ald6PRase by a partially purified kinase extract from peach leaves containing Ca2+-dependent protein kinases (CDPKs). Moreover, phosphorylation of recombinant Ald6PRase was inhibited by hexose-phosphates, phosphoenolpyruvate and pyrophosphate. We further show that phosphorylation of recombinant Ald6PRase was maximal using recombinant CDPKs. Overall, our results suggest that phosphorylation could fine-tune the activity of Ald6PRase.

Building a foundation for gene family analysis in Rosaceae genomes with a novel workflow: A case study in Pyrus architecture genes.

The rapid development of sequencing technologies has led to a deeper understanding of plant genomes. However, direct experimental evidence connecting genes to important agronomic traits is still lacking in most non-model plants. For instance, the genetic mechanisms underlying plant architecture are poorly understood in pome fruit trees, creating a major hurdle in developing new cultivars with desirable architecture, such as dwarfing rootstocks in European pear (Pyrus communis). An efficient way to identify genetic factors for important traits in non-model organisms can be to transfer knowledge across genomes. However, major obstacles exist, including complex evolutionary histories and variable quality and content of publicly available plant genomes. As researchers aim to link genes to traits of interest, these challenges can impede the transfer of experimental evidence across plant species, namely in the curation of high-quality, high-confidence gene models in an evolutionary context. Here we present a workflow using a collection of bioinformatic tools for the curation of deeply conserved gene families of interest across plant genomes. To study gene families involved in tree architecture in European pear and other rosaceous species, we used our workflow, plus a draft genome assembly and high-quality annotation of a second P. communis cultivar, 'd'Anjou.' Our comparative gene family approach revealed significant issues with the most recent 'Bartlett' genome - primarily thousands of missing genes due to methodological bias. After correcting assembly errors on a global scale in the 'Bartlett' genome, we used our workflow for targeted improvement of our genes of interest in both P. communis genomes, thus laying the groundwork for future functional studies in pear tree architecture. Further, our global gene family classification of 15 genomes across 6 genera provides a valuable and previously unavailable resource for the Rosaceae research community. With it, orthologs and other gene family members can be easily identified across any of the classified genomes. Importantly, our workflow can be easily adopted for any other plant genomes and gene families of interest.

Influence of Rosaceous Species and Driving Factors on Differentiation of Rhizospheric Bacteria in a Deciduous Broad-Leaved Forest.

Understanding plant-microbe interactions could provide the basis for improved phytoremediation, microbial resource utilization, and secondary metabolite production. Rhizosphere bacterial communities are strongly influenced by abiotic factors such as soil nutrient availability and the composition of such communities exhibits differentiation under different host plants. In a deciduous broad-leaved forest in Anhui Province, eastern China, the rhizospheric bacteria of three different tree species of the Rosaceae family (Sorbus alnifolia, Cerasus serrulata, and Photinia beauverdiana) were studied, with the bacteria of the bulk soil as controls. Bacterial community composition was determined using the Illumina platform for high-throughput sequencing of 16S rRNA genes. The results showed that the bacterial community composition varied between rhizospheric and bulk soils, and dominant bacterial phyla as Proteobacteria, Actinobacteria, and Acidobacteria were found in both soils. Information on predicted functional genes and pathways revealed significant differences between rhizospheric and bulk soil bacteria. It provided ample evidence for the different metabolic characteristics of the rhizosphere bacterial communities of the three tree species. Electrical conductivity (22.72%), total phosphorus concentration (21.89%), and urease activity (22%) were the main drivers for changes in the composition of the rhizosphere bacterial communities from the three tree species.

Characterization of transcriptomic response in ovules derived from inter-subgeneric hybridization in Prunus (Rosaceae) species.

Characterization of hybrid seed failure in Prunus provides insight into conserved or lineage-specific hybrid incompatibility mechanisms in plant species. Postzygotic hybrid incompatibility resulting from a cross between different species involves complex mechanisms occurring at various developmental stages. Embryo arrest, followed by seed abortion, is the first stage of such incompatibility reactions and inhibits hybrid seed development. In Prunus, a rosaceous woody species, some interspecific crosses result in fruit drop during the early stage of fruit development, in which inferior seed development may be accounted for the observed hybrid incompatibility. In this study, we investigated ovule development and the transcriptomes of developing ovules in inter-subgeneric crosses of Prunus. We conducted a cross of Prunus mume (subgenus Prunus), pollinated by P. persica (subgenus Amygdalus), and found that ovule and seed coat degeneration occurs before fruit drop. Transcriptome analysis identified differentially expressed genes enriched in several GO pathways, including organelle development, stimulus response, and signaling. Among these pathways, the organelle-related genes were actively regulated during ovule development, as they showed higher expression in the early stage of interspecific crosses and declined in the later stage, suggesting that the differential regulation of organelle function may induce the degeneration of hybrid ovules. Additionally, genes related to ovule and seed coat development, such as genes encoding AGL-like and auxin response, were differentially regulated in Prunus interspecific crosses. Our results provide histological and molecular information on hybrid seed abortion in Prunus that could be utilized to develop new hybrid crops. Additionally, we compared and discussed transcriptome responses to hybrid seed failure in Prunus and other plant species, which provides insight into conserved or lineage-specific hybrid incompatibility mechanisms in some plant species.

Effect of Host Plants on the Viability of Black-veined White Aporia crataegi L. at Low Natural Population Density

Patterns of interaction of an oligophagous insect, black-veined white Aporia crataegi L., with host plants in years with a low density of its natural population are analyzed. In years with low A. crataegi population abundance in the south of Sverdlovsk region (Sysertsky district), the species composition of caterpillar host plants gradually decreases from several woody rosaceous species of the genera Padus, Sorbus, Malus, and Crataegus to one species (Padus avium Mill.). It is shown that the mortality rate is higher for fifth-instar caterpillars developing on mountain ash than for those developing on bird cherry. Adult A. crataegi developing on bird cherry are not only larger but also grow faster than those developing on mountain ash, which may be an additional factor responsible for the observed chronographic variation during the emergence of A. crataegi generation in nature.

Micropropagation of Rosaceous Species SAM Grown in Temperate Climate.

The benefits of in vitro plant cultivation are mainly due to very high multiplication rate. Cultivation of plant material in vitro can be carried out during the whole year regardless of the time of the year or weather conditions. We create artificial conditions in the lab (heat, light, humidity), and we can regulate these conditions at any time. For the preservation of cultivar identity, we recommend establishing in vitro cultures from shoot tips usually larger than 0.2mm. In practice, in vitro cultivation of plants uses these growth regulators to achieve organogenesis, for example, root formation, prolonged growth, or multiplication. During each subculture, these cultures are then transferred on a solid agar medium in the form of actively growing multiple shoots with a well-differentiated shoot tip containing meristematic area. Cytokinins are important for cell division and causes branching of plants. Auxins, both endogenous and exogenous, act at as a trigger for the differentiation and formation of root primordia. Morphological characteristics (formation of leaves or callus) and shoot development should be observed during in vitro multiplication and after transfer to ex vitro conditions.

Hunting for novel disease resistance genes: observations and opportunities from the Rosaceae

Identifying effective genes for disease resistance is a bottleneck in plant improvement and requires resource-intensive phenotyping followed by molecular mapping, tagging, and (in some cases) cloning. Across plant species and pathogen types, the nucleotide binding-leucine rich repeat (NB-LRR) superfamily of genes has been repeatedly implicated in detection and response to pathogens, yielding a resistance phenotype. While NB-LRR genes can be identified from DNA sequence it is impossible to associate a specific NB-LRR gene with a cognate pathogen species or race based on sequence alone. Working in the Rosaceae, we have developed a robust, semi-automated informatics pipeline to identify NB-containing gene sequences from whole genome sequences and interpret evolutionary patterns across multiple species within the family. We have applied our comparative approach to 3 rosaceous genomes, those of apple, peach, and the woodland strawberry. Our analyses reveal 7 dominant NB gene lineages. With few exceptions, each lineage is present in each species, suggesting an ancient origin. While most NB lineages are found in each rosaceous species, species differ profoundly in terms of allelic diversity within each lineage. We are using patterns of allelic diversification across species to develop hypotheses that link DNA sequences with putative gene function. Paired with targeted sequencing of the NB gene component of genebank collections, our comparative analyses have the potential to identify plant genotypes with novel NB alleles for subsequent targeted phenotypic testing. Rose black spot (caused by Diplocarpon rosae Wolf) is a significant disease impacting landscape roses and creation of disease resistant cultivars is an important breeding goal. In the next phase of this project, we are applying our comparative genomics pipeline and targeted sequencing of the NB gene component of rose genotypes to identify markers linked to black spot resistance for marker assisted breeding.

Phylogenetic analysis of S-RNase-like genes from Fragaria provides evidence for ancestral S-RNase duplication in the Rosaceae lineage

Self-incompatibility (SI) is associated with cultural and breeding difficulties in rosaceous fruit trees such as cherry, plum, apple, and pear. Many species in Rosaceae, Solanaceae, and Plantaginaceae have S-RNase-based gametophytic SI, with S-RNase and F-box proteins as pistil S and pollen S determinants, respectively. Although the S-RNase-based gametophytic SI system is generally believed to have evolved once in a eudicot ancestor before the divergence of asterids and rosids, we previously found that S-RNase and its homologs were duplicated in the ancestral genome of Rosaceae. Here, we reinvestigated the evolutionary paths of S-RNase-like genes using homologs from the diploid Fragaria species F. nipponica (SI), F. nubicola (SI), and F. iinumae (SI phenotype undetermined). Our phylogenetic analysis and estimation of proxy ages for the establishment of S-RNase and its homologs in rosaceous species supports the previously inferred ancestral S-RNase duplication in the Rosaceae lineage. The divergence of S-RNases in subtribe Malinae and Prunus predated the most recent common ancestor of Rosaceae, which includes genes from three rosaceous species (Fragaria, Prunus, and Malus). The results of the present study strengthen the idea that the genus Prunus and subtribe Malinae have evolved different S-RNases for SI recognition. These results may explain the genus- or subtribe-specific SI recognition mechanism operating in rosaceous fruit trees and will be valuable for artificial SI control and self-compatible breeding.

Sexual intraspecific recombination but not de novo origin governs the genesis of new apomictic genotypes in Potentilla puberula (Rosaceae)

Apomixis - asexual reproduction via seeds - might arise de novo following polyploidisation events, or via reproductive transfer of apomixis. Both processes can be obtained within species or via hybridisation. We aimed to determine the origin of apomictic genotypes in Potentilla puberula, a rosaceous species showing reproductive differentiation with ploidy: sexual tetraploids and apomictic penta- to octoploids, which regularly co-occur in sympatry. The study is based on 726 individuals, comprising all cytotypes, collected from 138 populations in the Eastern European Alps. We established relationships of cytotypes based on AFLP fingerprinting and cpDNA sequencing to test (1) whether the apomicts are of recurrent allopolyploid origin or originated from within the species via autopolyploidy, and (2) whether there are indications for reproductive transfer versus de novo origin of apomixis. Three principal pathways were identified which explain the origin of new apomictic genotypes, all involving at least one apomictic parent and thus compatible with the idea of reproductive transfer of the apomictic trait to the progeny: (1) self-fertilisation of unreduced egg cells in apomicts; (2) cross-fertilisation among apomicts; and (3) occasionally, heteroploid crosses among sexuals and apomicts. Autopolyploids derived from tetraploid sexuals were repeatedly observed, but did not express apomixis. Finally, our results suggest no role of other species in the origin of extant apomictic genotypes of P. puberula, although local hybrids with P. crantzii were identified. In conclusion, our results show that the formation of new apomictic genotypes required a genetic contribution from at least one apomictic parent. This finding is in accordance with the idea that apomixis is inheritable in P. puberula. On the contrary, lack of apomixis in penta- and hexaploids derived from sexual backgrounds did not support the hypothesis of a de novo origin of apomixis. Relatively high frequency of remnant sexuality in the apomicts involving different cytological pathways of seed formation can explain their high cytological and genotypic diversity. Finally, lack of global introgression from a third taxon is in support of P. puberula as a concise, although highly diverse, species.

Variability in leaf wettability and surface water retention of main species in semiarid Loess Plateau of China

AbstractLeaf wettability, adhesion or repulsion of water drops, varies greatly among species and plays an important role in plant–soil hydrological relations. This study aimed to examine the variability in leaf wettability among species in different habitats and growth periods, and their relationships with plant surface water retention in the semiarid Loess Plateau of China. The leaf adaxial and abaxial contact angles, the surface water retention of leaves and individual plants, and general plant traits of 68 species belonging to 28 families were examined from May to August in 2017. Results showed that leaf water contact angles ranged from 27.3° to 133.4° and leaves with higher contact angles normally had lower variation coefficients. Leaf wettability was affected by internal properties (including leaf side, family, and leaf age) and external conditions (growth period), whereas the life form and slope aspect did not show significant effects. There were 47 species having higher contact angles on adaxial than abaxial surfaces, and the differences were significant in 23 species. Gramineous and leguminous species were more unwettable than compositae and rosaceous species. New leaves were more unwettable than old leaves. Surface wettability increased from May–June to July–August period. Leaf wettability was positively correlated with leaf surface water retention and was the best predictor of individual plant surface water retention compared with other plant traits. Leaf wettability showed interspecific differences associated with family and growth stage and can be a considerable variable in predicting canopy interception and evaluating vegetation hydrological function in drought environments.

Asymmetric reproductive interference: The consequences of cross-pollination on reproductive success in sexual-apomictic populations of Potentilla puberula (Rosaceae).

Apomixis evolves from a sexual background and usually is linked to polyploidization. Pseudogamous gametophytic apomicts, which require a fertilization to initiate seed development, of various ploidy levels frequently co‐occur with their lower‐ploid sexual ancestors, but the stability of such mixed populations is affected by reproductive interferences mediated by cross‐pollination. Thereby, reproductive success of crosses depends on the difference in ploidy levels of mating partners, that is, on tolerance of deviation from the balanced ratio of maternal versus paternal genomes. Quality of pollen can further affect reproductive success in intercytotype pollinations. Cross‐fertilization, however, can be avoided by selfing which may be induced upon pollination with mixtures of self‐ and cross‐pollen (i.e., mentor effects). We tested for reproductive compatibility of naturally co‐occurring tetraploid sexuals and penta‐ to octoploid apomicts in the rosaceous species Potentilla puberula by means of controlled crosses. We estimated the role of selfing as a crossing barrier and effects of self‐ and cross‐pollen quality as well as maternal: paternal genomic ratios in the endosperm on reproductive success. Cross‐fertilization of sexuals by apomicts was not blocked by selfing, and seed set was reduced in hetero‐ compared to homoploid crosses. Thereby, seed set was negatively related to deviations from balanced parental genomic ratios in the endosperm. In contrast, seed set in the apomictic cytotypes was not reduced in hetero‐ compared to homoploid crosses. Thus, apomictic cytotypes either avoided intercytotype cross‐fertilization through selfing, tolerated intercytotype cross‐fertilizations without negative effects on reproductive success, or even benefitted from higher pollen quality in intercytotype pollinations. Our experiment provides evidence for asymmetric reproductive interference, in favor of the apomicts, with significantly reduced seed set of sexuals in cytologically mixed populations, whereas seed set in apomicts was not affected. Incompleteness of crossing barriers further indicated at least partial losses of a parental genomic endosperm balance requirement.

Population genetic analysis of a global collection of Fragaria vesca using microsatellite markers.

The woodland strawberry, Fragaria vesca, holds great promise as a model organism. It not only represents the important Rosaceae family that includes economically important species such as apples, pears, peaches and roses, but it also complements the well-known model organism Arabidopsis thaliana in key areas such as perennial life cycle and the development of fleshy fruit. Analysis of wild populations of A. thaliana has shed light on several important developmental pathways controlling, for example, flowering time and plant growth, suggesting that a similar approach using F. vesca might add to our understanding on the development of rosaceous species and perennials in general. As a first step, 298 F. vesca plants were analyzed using microsatellite markers with the primary aim of analyzing population structure and distribution of genetic diversity. Of the 68 markers tested, 56 were polymorphic, with an average of 4.46 alleles per locus. Our analysis partly confirms previous classification of F. vesca subspecies in North America and suggests two groups within the subsp. bracteata. In addition, F. vesca subsp. vesca forms a single global population with evidence that the Icelandic group is a separate cluster from the main Eurasian population.

Complete plastid genome of Eriobotrya japonica (Thunb.) Lindl and comparative analysis in Rosaceae.

Eriobotrya japonica (Thunb.) Lindl (loquat) is an evergreen Rosaceae fruit tree widely distributed in subtropical regions. Its leaves are considered as traditional Chinese medicine and are of high medical value especially for cough and emesis. Thus, we sequenced the complete plastid genome of E. japonica to better utilize this important species. The complete plastid genome of E. japonica is 159,137 bp in length, which contains a typical quadripartite structure with a pair of inverted repeats (IR, 26,326 bp) separated by large (LSC, 89,202 bp) and small (SSC, 19,283 bp) single-copy regions. The E. japonica plastid genome encodes 112 unique genes which consist of 78 protein-coding genes, 30 tRNA genes and 4 rRNA genes. Gene structure and content of E. japonica plastid genome are quite conserved and show similarity among Rosaceous species. Five large indels are unique to E. japonica in comparison with Pyrus pyrifolia and Prunus persica, which could be utilized as molecular markers. A total of 72 simple sequence repeats (SSRs) were detected and most of them are mononucleotide repeats composed of A or T, indicating a strong A or T bias for base composition. The Ka and Ks ratios of most genes are lower than 1, which suggests that most genes are under purifying selection. The phylogenetic analysis described the evolutionary relationship within Rosaceae and fully supported a close relationship between E. japonica and P. pyrifolia. Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-016-3702-3) contains supplementary material, which is available to authorized users.

Systematic Analysis of the 4-Coumarate:Coenzyme A Ligase (4CL) Related Genes and Expression Profiling during Fruit Development in the Chinese Pear.

In plants, 4-coumarate:coenzyme A ligases (4CLs), comprising some of the adenylate-forming enzymes, are key enzymes involved in regulating lignin metabolism and the biosynthesis of flavonoids and other secondary metabolites. Although several 4CL-related proteins were shown to play roles in secondary metabolism, no comprehensive study on 4CL-related genes in the pear and other Rosaceae species has been reported. In this study, we identified 4CL-related genes in the apple, peach, yangmei, and pear genomes using DNATOOLS software and inferred their evolutionary relationships using phylogenetic analysis, collinearity analysis, conserved motif analysis, and structure analysis. A total of 149 4CL-related genes in four Rosaceous species (pear, apple, peach, and yangmei) were identified, with 30 members in the pear. We explored the functions of several 4CL and acyl-coenzyme A synthetase (ACS) genes during the development of pear fruit by quantitative real-time PCR (qRT-PCR). We found that duplication events had occurred in the 30 4CL-related genes in the pear. These duplicated 4CL-related genes are distributed unevenly across all pear chromosomes except chromosomes 4, 8, 11, and 12. The results of this study provide a basis for further investigation of both the functions and evolutionary history of 4CL-related genes.

Ploidy Levels, Relative Genome Sizes, and Base Pair Composition in Cotoneaster

The genus Cotoneaster (Rosaceae, Maloideae) is highly diverse, containing ≈400 species. Like other maloids, there is a high frequency of naturally occurring polyploids within the genus, with most species being tetraploid or triploid. Apomixis is also prevalent and is associated with polyploidy. The objective of this study was to estimate genome sizes and infer ploidy levels for species that had not previously been investigated as well as compare estimates using two fluorochromes and determine base pair (bp) composition. Chromosome counts of seven species confirmed ploidy levels estimated from flow cytometric analysis of nuclei stained with 4′,6-diamidino-2-phenylindole (DAPI). Monoploid (1Cx) genome sizes ranged from 0.71 to 0.96 pg. Differences in monoploid genome size were not related to current taxonomic treatment, indicating that while chromosome sizes may vary among species, there are no clear differences related to subgeneric groups. A comparison of DAPI and propidium iodide (PI) showed a difference in DNA staining in Cotoneaster comparable to other rosaceous species. Base pair composition (AT%) in Cotoneaster ranged from 58.4% to 60.8%, which led to overestimation of genome size estimates in many cases—assuming the estimates of the DNA intercalator are accurate. Our findings will inform breeders with regard to the reproductive behavior of potential parents and may be used to confirm hybrids from interploid crosses.

Development of retrotransposon-based molecular markers and their application in genetic mapping in chokecherry (Prunus virginiana L.)

Retrotransposons are the largest group of transposable elements (TEs) that are ubiquitous and well dispersed in plant genomes. Transposition/insertion of TEs on chromosomes often generates unique repeat junctions (RJs) between TEs and their flanking sequences. Long terminal repeats (LTR) are well conserved and abundant in plant genomes, making LTR retrotransposons valuable for development of TE junction-based markers. In this study, LTR retrotransposons and their RJs were detected from chokecherry genome sequences generated by Roche 454 sequencing. A total of 1246 LTR retrotransposons were identified, and 338 polymerase chain reaction primer pairs were designed. Of those, 336 were used to amplify DNA from chokecherry and other rosaceous species. An average of 283 of 336 (84.2 %) LTR primer pairs effectively amplified DNA from chokecherries. One hundred and seventeen chokecherry LTR primers also produced amplification in other Prunus (99) or rosaceous species (19). A total of 59 of 78 polymorphic LTR markers were qualified for linkage map construction according to the segregation distortion Chi-square (χ 2) test. Forty-eight LTR markers were successfully located on a previously constructed chokecherry map. The majority of the LTR markers were mapped on LG XI of the chokecherry map. Our results suggest that LTR marker development using random genome sequences is rapid and cost-efficient. Confirmed applicability of LTR markers in map construction and genetic mapping will facilitate genetic research in chokecherry and other rosaceous species.

A standard nomenclature for gene designation in the Rosaceae

The plant family Rosaceae is comprised of a highly diverse group of plants, with a wide range of ornamental and agricultural uses throughout the world. Comparative genomic analyses between rosaceous species are increasingly being used to assign putative function. With the wealth of new data, there is a need to standardize nomenclature to ensure uniformity and clarity across research groups. The Rosaceae Gene Name Standardization Committee was formed to develop a naming guideline for genes in Rosaceae family members. This manuscript details the issues involved with naming genes and proposes a common nomenclature system. The Genome Database for Rosaceae (GDR) ( www.rosaceae.org ) has developed a gene database that will support user-submitted gene names and annotations in addition to the predicted genes from whole genome sequences. GDR also hosts user-curated lists of gene class symbols. To facilitate standardization of gene names and avoid misinterpretation, the committee recommends that the GDR serve as the database of record for the names of Rosaceae genes and that gene names, gene class symbols, and gene annotation be registered in GDR prior to publication in peer reviewed journals.

Development, Characterization and Transferability of Peach Genic SSRs to some Rosaceae species

Aims: The present study was carried out to extract expressed sequence tags-simple sequence repeats (EST-SSRs) for peach (Prunus persica L.) and to study their transferability across some Rosaceae species such as apricot, apple, rose and strawberry. Study Design: Cross-transferability check of Prunus EST-SSRs to some Rosaceae species. Place and Duration of Study: Sample: Department of Biotechnology, Dr Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan-173230, Himachal Pradesh, India, between January 2012 and November 2012. Methodology: SSRs were identified in EST sequences using SSR Identification Tool (SSRIT) and PRIMER3 software was used to design primers which were used to carry out PCR. Data generated by PCR for all the genotypes was used to calculate transferability of Prunus persica EST-SSRs to other Rosaceae species under study. Data analysis was done through NTSYSpc Version 2.02h by creating dendrograms in all the species under study. Original Research Article Kaur et al.; AIR, 3(2): 165-180, 2015; Article no.AIR.2015.013 166 Results: 2000 EST sequences were downloaded from the National Centre for Biotechnology Information (NCBI) web site. Sequences were assembled into contigs and singletons. 124 sequnces were found to contain SSRs using SSRIT. Out of these 43 were used for primer designing using PRIMER3 software which were custom synthesized and used for polymorphism study in six genotypes of peach. Thirty-eight primers showed scorable amplification, 20 being polymorphic. These 20 polymorphic primers of peach were then further used to carry out transferability studies in apricot, apple, rose and strawberry revealing 50%, 95%, 95% and 45% transferability, respectively. Dendrograms were generated using NTSYS ver.2.02h. The coefficient values were found to range from 0.483 to 0.711 in six peach genotypes and 0.451 to 0.975 in all 22 genotypes under study. Conclusion: Thus it can be concluded from the present study that ESTs of peach produce high polymorphism in different Rosaceous species, indicating their cross-transferability to different members of Rosaceous family.

Plant Comparative and Functional Genomics.

Plant Comparative and Functional Genomics.

An inexpensive and rapid genomic DNA extraction protocol for rosaceous species

SummaryThe development of high-throughput DNA genotyping, as part of marker-assisted selection and other DNA-based applications, has led to the need for a cheap, rapid and simple method for extracting genomic DNA (gDNA) from plant material. Commonly available gDNA extraction protocols do not provide consistently good quality gDNA from a wide range of plant species, particularly those in families such as the Rosaceae, which contain variable concentrations of polysaccharides and phenolic compounds. Furthermore, although commercial DNA extraction kits are widely available, they are expensive for the preparation of large numbers of DNA samples. Here, we present an improved gDNA extraction method which we have tested on 12 different species from the family Rosaceae: apple, apricot, blackberry, strawberry, peach, sweet cherry, sour cherry, pear, plum, red raspberry, quince, and almond. The method employs a novel lysis buffer which is a crucial step in obtaining high quality gDNA. It does not require expensive or toxic reagents and can be performed in the open laboratory, rather than in an air-flow hood. The method is reliable and produces good quality DNA that can be used in a variety of DNA-based studies.