Polyploidy plays an important role in plant adaptation to biotic and abiotic stresses. Alterations of the ploidy in grapevine plants regenerated via somatic embryogenesis (SE) may provide a source of genetic variability useful for the improvement of agronomic characteristics of crops. In the grapevine, the SE induction process may cause ploidy changes without alterations in DNA profile. In the present research, tetraploid plants were observed for 9.3% of ‘Frappato’ grapevine somatic embryos regenerated in medium supplemented with the growth regulators β-naphthoxyacetic acid (10 µM) and N⁶-benzylaminopurine (4.4 µM). Autotetraploid plants regenerated via SE without detectable changes in the DNA profiles were transferred in field conditions to analyze the effect of polyploidization. Different ploidy levels induced several anatomical and morphological changes of the shoots and mature leaves. Alterations have been also observed in stomata. The length and width of stomata of tetraploid leaves were 39.9 and 18.6% higher than diploids, respectively. The chloroplast number per guard cell pair was higher (5.2%) in tetraploid leaves. On the contrary, the stomatal index was markedly decreased (12%) in tetraploid leaves. The observed morphological alterations might be useful traits for breeding of grapevine varieties in a changing environment.

Somatic embryogenesis (SE) provides alternative methodologies for the propagation of grapevine (Vitis spp.) cultivars, conservation of their germplasm resources, and crop improvement. In this review, the current state of knowledge regarding grapevine SE as applied to these technologies is presented, with a focus on the benefits, challenges, and limitations of this method. The paper provides a comprehensive overview of the different steps involved in the grapevine SE process, including callus induction, maintenance of embryogenic cultures, and the production of plantlets. Additionally, the review explores the development of high-health plant material through SE; the molecular and biochemical mechanisms underlying SE, including the regulation of gene expression, hormone signaling pathways, and metabolic pathways; as well as its use in crop improvement programs. The review concludes by highlighting the future directions for grapevine SE research, including the development of new and improved protocols, the integration of SE with other plant tissue culture techniques, and the application of SE for the production of elite grapevine cultivars, for the conservation of endangered grapevine species as well as for cultivars with unique traits that are valuable for breeding programs.

O. ficus-indica (prickly pear cactus) is an important forage and food source in arid and semiarid ecosystems and is the most important cactus species in cultivation globally. The high degree of apomixis in the species is a hindrance in plant breeding programs where genetic segregation is sought for the selection of superior genotypes. To understand if in ovulo embryo rescue could increase the proportion of zygotic seedlings, we compared the mature seed-derived seedlings with those regenerated from in vitro embryo rescue at 20, 25, 30, 35, and 40 post-anthesis days (PADs) in four Italian cultivars. The seedlings were classified as apomictic or zygotic based on molecular marker analysis using inter-sequence single repeat (ISSR) primers. Multiple embryos were recovered from all the cultured immature ovules, and plantlets were regenerated and acclimatized to the field post hardening, with success rates ranging from 62% (‘Senza spine’) to 83% (‘Gialla’). The level of polyembryony differed among cultivars and recovery dates, with the highest being ‘Rossa’, producing 4.8 embryos/ovule at 35 PADs, and ‘Gialla’, the lowest, with 2.7 at 40 PADs. The maximum number of embryos observed within a single ovule was 14 in ‘Trunzara bianca’. ISSR analysis revealed that ovule culture at 35 PADs produced the highest percentage of zygotic seedlings in all the cultivars, from 51% (‘Rossa’) to 98% (‘Gialla’), with a high genotype effect as well. Mature seeds produced much fewer seedlings per seed, ranging from 1.2 in ‘Trunzara bianca’ to 2.0 in ‘Rossa’ and a lower percentage of zygotic seedlings (from 14% in ‘Rossa’ to 63% in ‘Gialla’). Our research opens a pathway to increase the availability of zygotic seedlings in O. ficus-indica breeding programs through in ovulo embryo culture.

This study is the first approach to in vitro asymbiotic germination of two species of Sicilian threatened terrestrial orchids, Anacamptis longicornu and Ophrys panormitana. Seeds were collected in the wild and cultured in two different media—Orchimax medium (OM) and Murashige and Skoog (MS)—and exposed to different photoperiods and temperatures to evaluate the best conditions for the specific stages of development. The germination of A. longicornu was very high on OM (95.5%) and lower on MS medium (21.4%), whereas O. panormitana germinated only on OM medium, with significantly lower percentages (12.0%), compared with A. longicornu. This difference is caused by variation in quality and quantity of nutrients used, primarily by nitrogen source. The results show that temperature and photoperiod widely affect seed germination and development. Although further investigations on asymbiotic and symbiotic germination are needed for the improvement of conservation of Mediterranean terrestrial orchids, our results contribute to the conservation of this group of plants.

Several taxa of Cactaceae are endangered by overcollection for commercial purposes, and most of the family is included in the Convention on International Trade in Endangered Species of Fauna and Flora (CITES). Micropropagation may play a key role to keep the pressure off wild populations and contribute to ex situ conservation of endangered taxa. One of the limits of micropropagation is the species-specific requirement of plant regulators for each taxon and sometimes even for different genotypes. With the micrografting technique the rootstock directly provides the scion with the necessary hormonal requirements. In this paper we present data on in vitro grafting of Pelecyphora aselliformis Ehrenberg, an Appendix I CITES listed species critically endangered and sought after by the horticultural trade, on micropropagated Opuntia ficus-indica Miller. Apical and sub-apical scions of P. aselliformis were used to perform micrografting with a successful rate of 97 and 81 % respectively. Survival rate after ex vivo transfer was 85 %. We hypothesize that this method could be applied to other endangered, slow growing taxa of Cactaceae thus contributing to the conservation of this endangered family.