A Reliable Regeneration Method in Genome-Editable Bell Pepper ‘Dempsey’

A reliable regeneration technique is critical for the improvement of pepper traits in the genome editing era. Recently, we reported that peppers were successfully and specifically edited using CRISPR tools, CRISPR/Cas9 and CRISPR/Cas12a (LbCpf1). Although genome-editing tools can be applied to modify peppers at the cellular level, feasible pepper regeneration techniques have not been developed. Therefore, we studied a pepper regeneration protocol for Capsicum annuum L. ‘Dempsey’, a bell pepper species that has been proven to be genome-editable. Three explant types were used in this study, including the first leaves, cotyledons and hypocotyls of pepper seedlings. The shoot buds of the tested explants were produced using 8 mg/L 6-benzylaminopurine (BAP)- and 6 mg/L indole-3-acetic acid (IAA)-containing shoot induction medium (SIM). The first leaves of the ‘Dempsey’ seedlings showed an average shooting rate of 69.8%, whereas the hypocotyls and cotyledons had approximately 25.5% and 19.5% shooting rates, respectively. The regenerated ‘Dempsey’ plants exhibited no alterations in fruit and fertile seed phenotypes. Furthermore, the parent ‘Dempsey’ and progenies of the regenerants were cytogenetically stable with the same chromosome numbers (2n = 24). Therefore, this regeneration protocol enables the precise molecular breeding of ‘Dempsey’ peppers when coupled with CRISPR tools.

Understanding the cone scale in Cupressaceae: insights from seed-cone teratology in Glyptostrobus pensilis

Both wild-type and teratological seed cones are described in the monoecious conifer Glyptostrobus pensilis and compared with those of other Cupressaceae sensu lato and other conifers. Some Cupressaceae apparently possess a proliferation of axillary structures in their cone scales. In our interpretation, in Glyptostrobus each bract of both typical and atypical seed cones bears two descending accessory shoots, interpreted here as seed scales (ovuliferous scales). The primary seed scale is fertile and forms the ovules, the second is sterile and forms characteristic tooth-like structures. The bract and the two axillary seed scales are each supplied with a single distinct vascular bundle that enters the cone axis as a separate strand; this vasculature also characterises the descending accessory short shoots in the vegetative parts of the crown. In wild-type seed cones, the fertile seed scale is reduced to its ovules, and the ovules are always axillary. In contrast, the ovules of some of the teratological seed cones examined were located at the centre of the cone scale. An additional tissue found on the upper surface of the sterile lower seed scale is here interpreted as the axis of the fertile seed scale. Thus, the central position of the ovules can be explained by recaulescent fusion of the upper fertile and lower sterile seed scales. In several teratological cone scales, the ovules were enveloped by an additional sterile tissue that is uniseriate and represents an epidermal outgrowth of the fertile seed scale. Close to the ovules, the epidermis was detached from lower tissue and surrounded the ovule completely, except at the micropyle. These teratological features are potentially significant in understanding seed-cone homologies among extant conifers.

Shallots in Croatia – genetics, morphology and nomenclature

- The term 'shallot' in Croatia denotes three genetically and morphologically different, vegetatively reproduced relatives of the common onion, Allium cepa L., which are mainly traditionally cultivated for consumption and as a spice: A. cepa Aggregatum group, (2n = 2x = 16), A. × proliferum (Moench) Schrad. (2n = 2x = 16) and A. × cornutum Clementi ex Vis. (2n = 3x = 24). This paper reviews the results of studies of their morpho- -anatomical characteristics and genetic structure. Although all three taxa were determined as varieties of the common onion, only the shallot A. cepa Aggregatum group (syn. A. ascalonicum L.) belongs to that species. The shallot A. × proliferum represents a hybrid between the two closely related species, A. cepa and A. fistulosum L. The third form of shallot, A. × cornutum is a still incompletely understood triploid hybrid between A. cepa and one or two closely related Allium species, whose identity has not been fully elucidated. In contrast to shallot A. cepa Aggregatum group, which has normal meiosis and produces fertile seed, hybrid shallots A. × proliferum and A. × cornutum are sterile, and reproduce exclusively vegetatively by underground bulbs or bulbils from the inflorescence.

Serotiny in southern hemisphere conifers

Serotiny is a widespread trait in angiosperms in the southern hemisphere; however, it is less common in conifers and has been little examined in the only two genera of southern conifers (Callitris and Widdringtonia) that have serotinous cones. There is variation across the family in the size of cones, the amount of seed contained and the time over which the cones stay closed on the plant. Cones from most of the species were collected in the field and various morphometric measurements made including cone wet and dry weight, the number of seeds contained and their likely viability. Cones from a selection of species with different cone sizes were heated to increasing temperatures, to investigate the ability of cones to protect the contained seeds from heat. In comparison to the flowering plants, serotiny has developed comparatively recently in southern conifers (in the last 10–20 million years). In Widdringtonia, serotiny is relatively weak, whereas in Callitris, it varies from strong to non-existent. Cone size and fertile-seed production across the two genera varies and the number of fertile seeds produced is positively related to the size of the cone. In some species, there are sterile seed-like bodies. These may have developed to confuse seed predators, so fertile seeds have a better chance of survival. Larger (heavier) cones are more effective in protecting the contained seeds from the heat of fires than are smaller ones. There is no simple relationship between the cone size and type of environment occupied by the species. In regions where fire is unlikely, predictable but mild or completely unpredictable, the species tend to be non-serotinous. In temperate regions where hot fires are likely to have been a selective agent, the species tend to be more strongly serotinous, although fire is not essential to open the cones. The community and environment in which a species has evolved is likely to have influenced the development of the degree of serotiny for each species and this may still be a variable property among populations of some species, depending on the fire regime of the area in which they grow.

Rootstocks for Cherries from the Department of Fruit Science Budapest

Cherry rootstock breeding started at the Department of Fruit Science, SSU Budapest by the late 50-s and the activity can be divided into three main groups. In the first stage the activity was focused on collection of native mahaleb cherry (Prunus mahaleb L.) varieties lead by L. Sebők. After evaluation in the nursery and orchard tests there are four promising rootstock cultivars selected from this material: 'Korponay' used as self fertile seed tree, its seedlings are recommended for sour cherries. The mahaleb varieties 'Bogdány' (vigorous), 'Egervár' and 'Magyar' (medium vigour) are propagated by cuttings. The next project has started in 1979 with the aim to select self fertile mahaleb seed trees producing homogeneous seedling populations with reduced vigour. Inbred populations from isolated flowering self fertile trees were produced and planted out in 1980. The inbreeding of 'Korponay' self fertile P. mahaleb variety resulted in specimens with different fruit colour (yellow, red, black), fruit shape and size. From among them self fertile trees were selected with various growth characteristics. Seedlings of that self fertile mother trees (S2 population) were tested in seedbed, they showed homogeneous phenotype characteristics as liners in the nursery. As rootstock of 'Érdi bőtermő' sour cherry in the orchard most of the S2 lines proved to be less vigorous in comparison to SI popuplations. 'Érdi bőtermő' trees budded on certain S2 lines in the orchard are more productive than those on S1 ('Korponay' seedling). Characteristics of the S2 generation as seed tree were studied as well. We expect to get morphologically homogeneous seedling populations with different growth vigour and good productivity in the later inbred generations. In the last couple of decades the research activity concerning ground-cherry and its hybrids resulted in dwarfing rootstocks. Prunus fruticosa Pall. hybrids from the natural flora of Hungary were collected and artificial hybrids were created between P. fruticosa and mahaleb cherry. Most of them are in the initial tests, only one of them is before registration, named 'Prob', which is a dwarf rootstock for sweet cherry. By the screening of new hybrids medium vigorous or semi dwarfing and precocious rootstocks seem to be promising for the cherry industry.

Rootstocks for Cherries from the Department of Fruit Science Budapest

Cherry rootstock breeding started at the Department of Fruit Science, SSU Budapest by the late 50-s and the activity can be divided into three main groups. In the first stage the activity was focused on collection of native mahaleb cherry (Prunus mahaleb L.) varieties lead by L. Sebők. After evaluation in the nursery and orchard tests there are four promising rootstock cultivars selected from this material: 'Korponay' used as self fertile seed tree, its seedlings are recommended for sour cherries. The mahaleb varieties 'Bogdány' (vigorous), 'Egervár' and 'Magyar' (medium vigour) are propagated by cuttings. The next project has started in 1979 with the aim to select self fertile mahaleb seed trees producing homogeneous seedling populations with reduced vigour. Inbred populations from isolated flowering self fertile trees were produced and planted out in 1980. The inbreeding of 'Korponay' self fertile P. mahaleb variety resulted in specimens with different fruit colour (yellow, red, black), fruit shape and size. From among them self fertile trees were selected with various growth characteristics. Seedlings of that self fertile mother trees (S2 population) were tested in seedbed, they showed homogeneous phenotype characteristics as liners in the nursery. As rootstock of 'Érdi bőtermő' sour cherry in the orchard most of the S2 lines proved to be less vigorous in comparison to SI popuplations. 'Érdi bőtermő' trees budded on certain S2 lines in the orchard are more productive than those on S1 ('Korponay' seedling). Characteristics of the S2 generation as seed tree were studied as well. We expect to get morphologically homogeneous seedling populations with different growth vigour and good productivity in the later inbred generations. In the last couple of decades the research activity concerning ground-cherry and its hybrids resulted in dwarfing rootstocks. Prunus fruticosa Pall. hybrids from the natural flora of Hungary were collected and artificial hybrids were created between P. fruticosa and mahaleb cherry. Most of them are in the initial tests, only one of them is before registration, named 'Prob', which is a dwarf rootstock for sweet cherry. By the screening of new hybrids medium vigorous or semi dwarfing and precocious rootstocks seem to be promising for the cherry industry.