Tetraploids of daylily have taken a leading position among the daylily cultivars due to desirable traits such as vigorous growth and flowers with more intense colour. In our previous studies, several tetraploids of daylily cultivars ‘Blink of an Eye’ and ‘Berlin Multi’ were obtained using in vitro techniques with different antimitotic agents (colchicine, oryzalin, trifluralin, and amiprophos methyl). The purposes of this study were to evaluate changes in daylily tetraploids in relation to their diploid counterparts and to assess variation among diploids and tetraploids derived from genetically homogenous plant material treated with antimitotic agents. In the first year of the ex vitro cultivation, growth of tetraploids was poorer in comparison with diploids, but in the second year, tetraploid growth was much more vigorous. Compared to diploids, in tetraploids of both cultivars percentage of flowering plants was lower, flowering was delayed by 8 days and 1 month in ‘Blink of an Eye’ and ‘Berlin Multi’, respectively, and bud number per scape was lower by approximately 20 and 40 %, respectively. Tetraploid leaves and flowers of both cultivars were significantly larger, chlorophyll concentration index was higher by approximately 40 %, and stomata were longer by 35 %. In ‘Berlin Multi’ tetraploids, inflorescence stems were shorter by 20 %. In ‘Blink of an Eye’, variation in flower colour tone and shape and stamen malformation rate was detected both within the unconverted diploids and tetraploids but was more evident in tetraploids. In both cultivars, variation was also found in the nuclear DNA content, which ranged in diploids and tetraploids, respectively, 8.02–8.53 and 16.01–17.13 pg in ‘Berlin Multi’ and 8.28–8.71 and 15.93–17.36 pg in ‘Blink of an Eye’. Since the variation, while less evident, also occurred in the diploids (regenerated from the antimitotic treated material), we suppose that these variations could be due to antimitotic agents that can induce not only chromosome doubling but also chromosomal and gene mutations. The extent and character of these changes can be related to parental genotype and/or antimitotic agent. Further research is required at the cytological and molecular level to explain the character of changes, epigenetic and/or genetic.

This work focuses on the achievements of Polish researchers in the field of vegetative reproduction of plants under in vitro conditions. For more than 50 years, micropropagation methods have been used in Poland whenever vegetative reproduction is necessary. Most perennial horticultural crops, such as fruit plants, the majority of ornamental geophytes, and some vegetables (e.g., rhubarb and horseradish), require clonal reproduction owing to their high heterozygosity, i.e., their offsprings when reproduced from seeds, do not repeat the parental characteristics. Various goals have been pursued in the development of regenerative and in vitro propagation systems for each of the aforementioned groups of plants, such as pathogen elimination, reproduction of healthy plants, rapid multiplication of newly obtained valuable breeding lines and cultivars, and breeding to obtain polyploids, haploids, and doubled haploids. Owing to the growing interest of researchers and plant producers in environmentally friendly technologies, one of the sections is devoted to the issue of biotization of micropropagated plants.

Polyploidization is an important source of variability for plant breeding. Polyploids are often characterised by increased resistance to biotic and abiotic stresses. Since drought and pathogen attack are the main threats to apple cultivation, obtaining new sources of resistance is an important issue for apple breeding. The newly obtained autotetraploid clones of apple cv. ‘Redchief’ showed superior resistance to fire blight. The aim of the presented research was the in-depth phenotypic characterisation of ‘Redchief’ tetraploids and assessment of their response to drought at the physiological and genetic level. The growth of own-rooted five-year-old trees of ‘Redchief’ tetraploids was poor compared with diploids; all growth parameters—the number and length of current season shoots, the total length of current season shoots per tree and the cross-section area of the trunk—were reduced in tetraploid clones. Grafting on M9 rootstock improved the growth characteristics of ‘Redchief’ tetraploids. Compared with diploid plants, the leaves of tetraploids were thicker, with altered shape, higher chlorophyll content, and larger stomata, but the stomatal density decreased. The leaf anatomical structure of tetraploids was changed, the adaxial and abaxial epidermis and both types of mesophyll were significantly thicker than in diploids. Moreover, the pollen grains of tetraploids were larger, but their viability and germination were reduced. Under conditions of limited water supply, the reduction in growth parameters was smaller and the physiological parameters were higher in the ‘Redchief’ tetraploid clone 4x-25 than in diploid plants. The expression of APX gene was higher in tetraploids than in diploids 15 days after drought stress induction. The results suggest the enhanced drought tolerance of the studied ‘Redchief’ autotetraploid clone compared with its diploid counterpart.

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.

There are substantial variations in bulbing (bulb formation) efficiency among micropropagated tulip cultivars. The mechanisms involved are poorly understood, but presumably involve cytokinins (CKs) for several reasons. Therefore, we explored CK profiles and dynamics in ‘Blue Parrot’ and ‘Prominence’ cultivars (which have low and high bulbing efficiency, respectively) during the in vitro propagation stages: the last shoot multiplication subculture extended to 14 weeks (S1–S2), the shoot cooling at 5 °C for induction of bulb formation (S3–S4) and the bulb growth initiation after the end of cooling (S5–S6). The CK thidiazuron (TDZ) is routinely used in tulip micropropagation at the shoot multiplication stage, but replacing it with isopentenyladenine (iP) during the last multiplication subculture substantially changed CK dynamics in later stages, and significantly increased bulb formation rates in both cultivars. Generally, the most abundant CKs in both cultivars were the isoprenoid CK types, trans-zeatin (tZ), iP, cis-zeatin and dihydrozeatin. However, ‘Prominence’ shoots had much lower cis- to trans-Z-type CK ratios than ‘Blue Parrot’ shoots, and generally higher levels of physiologically active CKs (free bases tZ, iP and their ribosides) until the last phase of bulb formation, S6 (bulb growth initiation, i.e. swelling of shoot bases), 6 weeks after the end of cold treatment. In this phase total active CK and O-glucoside contents sharply declined in ‘Prominence’ shoots, but not in ‘Blue Parrot’ shoots pretreated with iP. In contrast, the low bulbing ability observed in ‘Prominence’ shoots pretreated with TDZ and ‘Blue Parrot’ shoots pretreated with either TDZ or iP was associated with a gradual rise in active CK and O-glucoside contents after the end of cooling. The results suggest that low bulbing efficiency may be related to down-regulation of tZ biosynthesis, and high bulbing efficiency to a transient increase in active CK forms (mainly tZs) in response to cold treatment during the bulb induction phase, S4 (at the end of cold treatment), followed by a rapid decrease during bulb formation, S6 (6 weeks after the end of cooling).