Production of Homozygous Red Beet (Beta vulgaris L. subsp. vulgaris) Plants by Ovule Culture.

Hybrid varieties dominate the red beet market. The breeding process necessary to produce these cultivars is very difficult and time consuming. The application of in vitro gynogenesis can reduce the time needed to produce the corresponding homozygous pure lines to a few months. Our research team has developed a method to obtain red beet doubled haploid plants by gynogenesis. The best medium for gynogenesis induction is the B5 medium with the addition of 0.5 mg/L IAA, 0.2 mg/L BA, and 322 mg/L putrescine, whereas the best medium for shoot induction from embryos proved to be the MS medium supplemented with 0.1 mg/L NAA, 0.1 mg/L BA, and 0.5 mg/L putrescine. The shoots obtained were rooted on MS medium containing half the concentration of microelements and 3 mg/L NAA, 160 mg/L putrescine, and 20 g/L sucrose. Ploidy evaluation of gynogenetic plants was performed by flow cytometry and homozygosity or heterozygosity was determined by two isoenzymatic systems: PGI and AAT.

Pomological Characteristics and Ploidy Levels of Japanese Plum (Prunus salicina Lindl.) Cultivars Preserved in Poland.

Research on the resistance to frost, susceptibility to sharka, flowering biology, fruit setting, yield, and ploidy levels of 36 Japanese plum cultivars (mostly hybrids of Prunus salicina with Prunus cerasifera) were carried out in 2015-2020 at the Experimental Orchard located in Dabrowice near Skierniewice. Relatively mild winters with sporadic temperature drops to nearly -21 °C in January of 2017 and 2018 caused slight damage to several cultivars of Japanese plum insufficiently resistant to frost. The trees of most cultivars remained healthy, with no signs of damage. 'Barkhatnaya' and 'Tatyana' cultivars turned out to be very susceptible to sharka. 'Herkules' trees were the most vigorous. 'Barkhatnaja', 'Blue Gigant', 'Shater', and 'Tatyana' trees were characterized by weak growth. The trees of Japanese plum started flowering early, usually in the first or second decade of April. Most of the cultivars belonged to early season cultivars, the fruits of which ripened in July. Based on the assessment of tree productivity, 'Barkhatnaya', 'Inese', 'Shater', 'Tatyana', and 'Vanier' are the best for growing in the climate of Central Europe. 'Tsernushka', 'Chuk', 'Dofi Sandra', 'Early Golden', 'Ewierch Rannyj', 'Yevraziya', 'Gek', 'General', 'Kometa', 'Kometa Late', 'Maschenka', and 'Naidyona' trees also yielded well. 'Blue Gigant', 'Black Amber', and 'Herkules' had the largest fruits, and 'Chuk' and 'Inese' cultivars produced the smallest fruits. Among the assessed Japanese plum cultivars, those with round fruit, dark skin with various shades of purple, yellow flesh, and A cytometric analysis showed that almost all cultivars are diploid, except for 'Herkules' (possibly pentaploid) and 'Yevraziya' (possibly hexaploid or aneuploid).

Apple Autotetraploids with Enhanced Resistance to Apple Scab (Venturia inaequalis) Due to Genome Duplication-Phenotypic and Genetic Evaluation.

Among the fungal diseases of apple trees, serious yield losses are due to an apple scab caused by Venturia inaequalis. Protection against this disease is based mainly on chemical treatments, which are currently very limited. Therefore, it is extremely important to introduce cultivars with reduced susceptibility to this pathogen. One of the important sources of variability for breeding is the process of polyploidization. Newly obtained polyploids may acquire new features, including increased resistance to diseases. In our earlier studies, numerous tetraploids have been obtained for several apple cultivars with 'Free Redstar' tetraploids manifesting enhanced resistance to apple scab. In the present study, tetraploids of 'Free Redstar' were assessed in terms of phenotype and genotype with particular emphasis on the genetic background of their increased resistance to apple scab. Compared to diploid plants, tetraploids (own-rooted plants) were characterized with poor growth, especially during first growing season. They had considerably shorter shoots, fewer branches, smaller stem diameter, and reshaped leaves. In contrast to own-rooted plants, in M9-grafted three-year old trees, no significant differences between diplo- and tetraploids were observed, either in morphological or physiological parameters, with the exceptions of the increased leaf thickness and chlorophyll content recorded in tetraploids. Significant differences between sibling tetraploid clones were recorded, particularly in leaf shape and some physiological parameters. The amplified fragment length polymorphism (AFLP) analysis confirmed genetic polymorphism of tetraploid clones. Methylation-sensitive amplification polymorphism (MSAP) analysis showed that the level of DNA methylation was twice as high in young tetraploid plants as in a diploid donor tree, which may explain the weaker vigour of neotetraploids in the early period of their growth in the juvenile phase. Molecular analysis showed that 'Free Redstar' cultivar and their tetraploids bear six Rvi genes (Rvi5, Rvi6, Rvi8, Rvi11, Rvi14 and Rvi17). Transcriptome analysis confirmed enhanced resistance to apple scab of 'Free Redstar' tetraploids since the expression levels of genes related to resistance were strongly enhanced in tetraploids compared to their diploid counterparts.

Micropropagation of tulip: production of virus-free stock plants.

We describe here a new tulip micropropagation method based on the cyclic shoot multiplication in presence of the thidiazuron (TDZ), which enables the production of virus-free stock plants, speeds up breeding, and provides new genotypes for the market. In our novel protocol, cyclic shoot multiplication can be performed for 2-3 years by using TDZ instead of other cytokinins, as 6-benzylaminopurine (BAP) and N(6)-(-isopentyl)adenine (2iP). It makes possible to produce 500-2,000 microbulbs from one healthy plant. There are six main stages of tulip micropropagation. Stage 0 is the selection of true-to-type and virus-free plants, confirmed by ELISA. Fragments of flower stems isolated from bulbs are used as initial explants. Shoot multiplication is based on the regeneration of adventitious shoots, which are sub-cultured every 8 weeks. In the Stage 3, the specially prepared shoots are induced by low temperature treatment to form bulbs which finally develop on a sucrose-rich medium at 20 degrees C. Bulbs are then dried for 6 weeks and rooted in vivo. The number of multiplication subcultures should be limited to 5-10 cycles in order to lower the risk of mutation. Virus indexing should be repeated 3-4 times, at the initial stage and then during shoot multiplication. Genetic stability of micropropagated shoots can be confirmed using molecular markers.