After replanting apple (Malus domestica Borkh.) on the same site severe growth suppressions, and a decline in yield and fruit quality are observed in all apple producing areas worldwide.

Apple replant disease (ARD) is a soil-borne disease, which is of particular importance for fruit tree nurseries and fruit growers. The disease manifests by a poor vegetative development, stunted growth, and reduced yield in terms of quantity and quality, if apple plants (usually rootstocks) are replanted several times at the same site. Genotypespecific differences in the reaction of apple plants to ARD are documented, but less is known about the genetic mechanisms behind this symptomatology. Recent transcriptome analyses resulted in a number of candidate genes possibly involved in the plant response. In the present study, the expression of 108 selected candidate genes was investigated in root and leaf tissue of four different apple genotypes grown in untreated ARD soil and ARD soil disinfected by g-irradiation originating from two different sites in Germany. Thirty-nine out of the 108 candidate genes were differentially expressed in roots by taking a p-value of < 0.05 and a fold change of > 1.5 as cutoff. Sixteen genes were more than 4.5-fold upregulated in roots of plants grown in ARD soil. The four genes MNL2 (putative mannosidase); ALF5 (multi antimicrobial extrusion protein); UGT73B4 (uridine diphosphate (UDP)-glycosyltransferase 73B4), and ECHI (chitinbinding) were significantly upregulated in roots. These genes seem to be related to the host plant response to ARD, although they have never been described in this context before. Six of the highly upregulated genes belong to the phytoalexin biosynthesis pathway. Their genotype-specific gene expression pattern was consistent with the phytoalexin content measured in roots. The biphenyl synthase (BIS) genes were found to be useful as early biomarkers for ARD, because their expression pattern correlated well with the phenotypic reaction of the Malus genotypes investigated.

The generation of transgenic apple plants relies on the molecular analysis of transgene integration and expression based on polymerase chain reaction (PCR) analysis, blotting techniques and enzymatic assays on vitro leaves of putative transgenic regenerates. In order to assess the uniformity and the stability of transfer DNA (T-DNA) integration and gene expression, we studied 26 transgenic apple lines carrying the attacin E

Knowledge of pollen movement and frequency of interspecific hybridization in fragmented populations of rare species is a prerequisite for the implementation of conservation measures. In a large-scale study area (14,000 hectares) we analysed 297 Malus sylvestris trees with nine nuclear microsatellite markers. After open pollination of 564 offspring from 51 mother trees located in seven harvesting sites were investigated and genetic paternity analysis was performed. The paternal parent was identified for 213 offspring and the pollen dispersal distances between mother and pollen source were calculated. A large proportion of detected pollination events (42.4%) were observed within a radius of 50 m of the mother tree. The comparison of different tree densities indicated that with decreasing density the pollen dispersal distances increase. We observed pollination over long distances with a maximum of 10.7 km which is probably one of the reasons for a low spatial genetic structure within the M. sylvestris population and a stable genetic diversity in the offspring. Incorporating microsatellite data of 21 apple cultivars, a hybridization frequency of nearly 8% was determined. With decreasing tree density the number of hybridization events increased. Based on the results of our study an enhancement of the density of existing M. sylvestris populations is recommend to reduce the likelihood of hybridization. The production of young plants originated from seeds collected after open pollination is not advisable. Instead of that the seedlings for further reintroduction measures should be produced by controlled crossings in seed orchards to ensure ‘true type’ M. sylvestris individuals.

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