DNA-based S-genotyping of Japanese plum and pluot cultivars to clarify incompatibility relationships

Diploid japanese plum (Prunus salicina Lindl.) cultivars are commonly self-incompatible. To date, 14 incompatibility alleles (S-alleles) have been identified and labeled with alphabetical (S-a-S-n) and 5 with numeric codes (S-1, S-3-S-6). We applied polymerase chain reaction amplification of the S-RNase alleles with degenerate and allele-specific primers in 10 japanese plum cultivars and two pluots of unknown incompatibility alleles. Besides DNA sequencing, an additional method for the exact length determination of the first intron region was used for the first time for S-genotype japanese plums. The S-3-allele was shown to correspond to S-k in the alphabetic nomenclature, S-4 to S-c, S-5 to S-e, and S-6 to S-f. The S-5-allele-specific primer can be used as a reliable marker for self-compatibility in japanese plum. 'Black Amber', 'October Sun', 'TC Sun', and 'Super Giant' share the SbSc genotype, which was confirmed by test crosses. These cultivars belong to the widest incompatibility group currently known in japanese plum. An additional incompatibility group (ScSh) was established, including 'Green Sun' and 'Queen Rosa', a cultivar formerly known as a universal donor. By incorporating all previous and recent results, a table was assembled including 49 cultivars assigned to I-VII incompatibility groups, to the self-compatible group and to the group O of unique genotypes. These data may considerably contribute to further growing and breeding activities

Molecular analysis of S-haplotypes in peach, a self-compatible Prunus species

The most commercially grown peach [Prunus persica (L.) Batsch.] cultivars do not require cross-pollination for reasonable fruit set; however, self-incompatibility is a well-known feature within the Prunoideae subfamily. Isoelectric focusing and native polyacrylamide gel electrophoresis of S-ribonucleases; PCR analyses of S-RNase and S-haplotype-specific F-box genes as well as DNA sequencing were carried out to survey the self- (in)compatibility allele pool and to uncover the nature of self-compatibility in peach. From 25 cultivars and hybrids with considerable diversity in phenotype and origin, only two S-haplotypes were detected. Allele identity could be checked by exact length determination of the PCR-amplified fragments and/or partial sequencing of the peach S-1-, S-2-, and Prunus davidiana (Carr.) Franch. S-1 RNases. S-RNases of peach were detected to possess ribonuclease activity, and a single nucleotide polymorphism in the S,-RNase was shown, which represents a synonymous substitution and does not change the amino acid present at the position in the protein. A 700-bp fragment of the peach SFB gene was PCR-amplified, which is similar to the fragment size of functional Prunus L. SFBs. All data obtained in this study may support the contribution of genes outside the S-locus to the self-compatible phenotype of peaches

Molecular background of self-incompatibility in apricot

Prunus species show gametophytic self-incompatibility, a trait encoded by a single multiallelic locus, termed S-locus. The S-gene products in styles are pistil ribonucleases, while the pollen component is an F-box protein suggesting that the ubiquitin-mediated protein degradation system has a pivotal role in self/non-self recognition. Using non-equilibrium pH gradient electrophocusing and PCR analysis with several S-gene specific degenerate primer pairs allowed to identify nine new alleles in apricot. Their schematic structure was determined and confirmed to be different from the previously described alleles. Monitoring of different self-(in)compatibility alleles were also successful using a combination of the two techniques. Preliminary results for the molecular background of fruit set behaviour in apricot are discussed

Artificial frost treatment methods with stone fruits

Routine application of artificial frost treatment needs a detailed, exact technology, which would ensure the reliability of the results obtained. Buds being on spurs loose their frost resistance sooner because of their quick development. According to these results to examine samples containing both types of buds can lead to significant mistakes. Considering our experiences it seems that a sample containing 200 buds provides the reliable correctness. Further increasing of sample size decreases the value of deviation but it is not proportional to the work needed for the experiment. The following treatments are suggested: deep dormancy phase: -24 - -26°C, directly after deep dormancy:-21 - -24°C. beginning of February: -19 - -21°C, two weeks before blossoming -11- -12°C. two -days before blossoming: -4 - -6°C

Genetikai diverzitás vizsgálatok őshonos közép-európai Pinus cembra L. populációknál: populáció-ökológiai, populáció-genetikai, erdészeti, táj-, és vegetációtörténeti vonatkozások. = Genetic diversity studies on native Central-European populations of Pinus cembra L.: ecological, genetic, forestry, landscape-, and vegetation historical correlates

Genetikai vizsgálatokat végeztünk az Európában védett cirbolyafenyő populációinál. A vizsgálatok elsősorban a Kárpátok populációira terjedtek ki, majd a Kárpátok populációit összehasonlítottuk a faj központi areaján található, svájci Alpok populációival is. A Kárpátokból 10, az Alpokból 19 populációt vizsgáltunk meg kloroplasztisz DNS SSR markerekkel. 7 primert teszteltünk, ebből 5 variábilis volt. Az Alpok állományával való összehasonlítást 3 primerrel végeztük el. A vizsgált populációk genetikai diverzitása általánosan magas volt. Legvariábilisabb a Retyezát populációi és a szlovákiai Tátra állománya. Genetikailag legszegényebbek a Radnai havasok állománya illetve a Cindrel hegység kispopulációja. A Kárpátok populációi, bár egyedszámukat tekintve sokkal kisebbek, esetenként nagyobb genetikai variabilitást őriztek meg mind az Alpok populációi! A populációk közötti divergencia mértéke nagyobb volt a Kárpátokban mint az Alpokban. A két földrajzi terület (Alpok?Kárpátok) genetikailag elkülönült egymástól, de régiókon belül a genetikai- és a földrajzi távolság között nem volt szignifikáns korreláció. Vélhetőleg, a fragmentálódás óta eltelt idő rövid volt ahhoz, hogy a markáns genetikai különbségek jelentkezzenek. Vizsgálataink rámutattak arra, hogy a Kárpátok populációi értékes genetikai bázist képviselnek, amely elkülönül az Alpok populációitól. Felhívjuk a figyelmet ezen populációk védelmére, mert ez elengedhetetlen záloga lehet a faj túlélésének. | According to the aim of our study genetic analysis were made on Pinus cembra, protected species of the European forest ecosystems. Our study was primarily focused on the genetic diversity of populations from the Carpathians, later we compared Carpathian populations with those from the Swiss Alps, representing the central part of the species area. We included in our study 10 populations from the Carpathians and 19 populations from the Alps. 7 chloroplast SSR markers (simple sequence repeats) were tested, 5 worked and show variability. Diversity of populations, fixation indices (Fst) were calculated. Despite the peripheral locations and a presumed high degree of isolation, Carpathian populations have been able to maintain high diversity level! This is most likely owing to their high outcrossing rate and extensive gene flow. The highest diversity values exhibit populations from the Retezat Mountains and from the Slovakian Tatra, lowest values we found in the Rodnei mountains and Cindrel. The differentiation was lower among populations of the Alps compared to the populations of the Carpathians. Our study proved that Carpathian populations including the Tatra are valuable because they still preserve a rich gene-stock, different that from theAlps. Therefor we think its really important to focus on the Carpathians and to protect all these populations, because they will contribute esentially to the survival of the species along the European range

Genetic Diversity Within and Among Populations of Roseroot (Rhodiola rosea L.) Based on Molecular Markers

Rhodiola rosea L. is a perennial adaptogenic medicinal plant found in cool climate of the northern hemisphere. The species is very diverse both in terms of morphological characteristics and in the content of the pharmacologically active substances. The genetic diversity of four geographically distant roseroot populations was studied with ISSR and SSR markers. Using 7 ISSR primers 64 DNA fragments were generated and 85,94% of those were found to be polymorphic, indicating high genetic variability at the species level (gene diversity = 0.33, Shannon index = 0.48). Lower level of diversity was detected at the population level (Shannon-index ranged from 0.2173 to 0.2696). Only four out of the eight SSR markers used were informative during this study. The primer pairs for these four SSR markers produced 25 fragments with an average of 6.25 putative alleles per locus. Observed heterozygosity ranged from 0.4 to 1.0, whereas expected heterozygosity ranged from 0.47 to 0.84. Cluster analysis based on both markers revealed the same groups, individuals clustered according to their geographic origin. The Southern-Uralian population was the most genetically isolated. ITS analysis was used for the determination whether these Southern-Uralian individuals belong to the same species