The present study was conducted to establish a protocol for the regeneration of virus-free garlic plants through somatic embryogenesis of two Croatian garlic ecotypes. Basal parts of cloves from mother plants were cultured on a full Murashige and Skoog (MS) or modified MS medium (¼ of KNO3 and NH4NO3 and 2xMgSO4) containing 0.1 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) or 1 mg L−1 2,4-D + 0.5 mg L−1 kinetin (Kin) and representing four different treatments. Plants were regenerated in MS medium containing 0.1 mg L−1 2,4-D and rooted in a medium containing 0.05 mg L−1 1-naphthaleneacetic acid (NAA) + 0.005 mg L−1 6-(γ,γ-dimethylallylamino)purine (2iP). The presence of viruses (i.e., sanitary status) of the mother plants and regenerants was checked by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The mother plants were infected with onion yellow dwarf virus (OYDV) and leek yellow stripe virus (LYSV). In addition, the presence of garlic common latent virus (GCLV) was confirmed in four mother plants. Embryogenic callus developed in all four treatments with success ranging from 55% to 81% depending on treatment and ecotype. Plant conversion was significantly higher in somatic embryos developed in media containing 0.1 mg L−1 2,4-D than those developed in media containing 1 mg L−1 2,4-D + 0.5 mg L−1 Kin. Virus elimination success ranged from 13.3% up to 62.5% depending on garlic ecotype and treatment. The overall rate of virus elimination by somatic embryogenesis for both treatments and ecotypes were 20.7%, 22.9%, and 30.5% for OYDV, GCLV, and LYSV, respectively. Based on these results, somatic embryogenesis has been shown to be equally or more successful in eliminating garlic viruses compared to other in vitro methods.
Caper (Capparis orientalis Veill.) is a species rich in bioactive compounds, with positive effects on human health. It has a great adaptability to harsh environments and an exceptional ability to extract water from dry soils. In Croatia, the caper grows as a wild plant, and its cultivation is insignificant, which is probably due to propagation difficulties. Micropropagation could be a solution for this. The aim of this study was to investigate the success of the micropropagation, in vitro rooting, and acclimatization of Capparis orientalis Veill. Shoot proliferation was tested in a Murashige and Skoog (MS) medium, with sucrose or glucose, and in 13 treatments, presenting the combined effect of different cytokinins and their concentrations. The success of rooting was examined in relation to the impact of various auxins, durations of rooting, and carry-over effects. A better proliferation was achieved when sucrose was used. The highest number (18) of shoots/explants was obtained in the medium supplemented with 0.6 mg·L−1 meta-topolin, while the rooting was equally efficient in the media supplemented with 2 mg·L−1 of indole-3-acetic acid or indole-3-butyric acid, or in hormone-free rooting medium. A prolonged time in the media increased the rooting efficiency, while the carry-over effect had no influence. The acclimatization rate reached 66%. Additional efforts should be made to find out how to speed upthe rooting and enhance the acclimatization rate of caper grown in Croatia.
Leaf samples of 226 cultivated olive trees were collected from traditionally managed olive orchards and genotyped with eleven consensual SSR markers. The proportion of shared allele distance was used for the estimation of distances between olive genotypes. Cluster analyses were performed using a Fitch–Margoliash least-squares algorithm. The number of different genetic subgroups of olive genotypes (K) was investigated using STRUCTURE analysis. The standardization of allele lengths was performed to enable the comparison SSR profiles of Croatian olive genotypes with olive profiles obtained with the same SSR primers in OleaDB and WOGB databases. Overall, 73 SSR profiles of known Croatian varieties and 53 profiles of unknown olive genotypes were differentiated. Synonyms were detected in 18 varieties, and we found intra-varietal differences in 15 varieties. Three genetic subgroups of olive genotypes were determined. Following allele length standardization achieved using nine referral samples, the genetic profiles of 126 cultivated olive genotypes were compared to OleaDB and WOGB databases, out of which 92 genotypes were found to be unique to Croatian olive germplasm. The results revealed the wide genetic diversity of olive germplasm beyond the known, registered varieties. The FAZ_oliveDB database containing the profiles of 126 Croatian olive genotypes was created and made available for public use.
Fragaria nilgerrensis Schlecht. (wild strawberry) is widely distributed in Southwest China, characterized by stress tolerance and the fruits of a notable peach aroma. So far there is only limited knowledge of variability and genetic structure in this species. Using AFLP markers, we investigated the genetic variability of 37 plants of F. nilgerrensis sampled in six main mountain areas of Sichuan Province and analyzed their genetic structure. Genetic similarity according to Nei and Li was used for cluster analysis based on UPGMA method and Agglomerative Hierarchical Clustering. Stratification of plants into more distinctive genetic groups was determined using Bayesian structure analysis. Six primer combinations produced a total of 1302 fragments of which 818 (62.8%) were polymorphic. Bayesian analysis showed the 37 plants of F. nilgerrensis grouped into five distinctive genetic groups. Most of the plants from the same mountain area clustered into the same genetic group, indicating each area as an area with the unique genetic profile. The genetic parameters analyzed here indicate a huge genetic variability of F. nilgerrensis in Sichuan Province. Our results provide reference data for surveying and protecting the germplasm resources of F. nilgerrensis that could be used in strawberry breeding programs.