Somatic embryogenesis of Vitis vinifera L. (cv. Sugraone) from stigma and style culture

Somatic embryo and plant regeneration were induced from stigma and style culture of grapevine (Vitis vinifera L. cv. Sugraone). To obtain somatic embryogenesis, explants were cultured on Nitsch and Nitsch basal medium (NN) supplemented with 88 mM sucrose and various combinations of the auxin β-naphthoxyacetic acid (NOA, 0-10 μM) and the cytokinin 6-benzylaminopurine (BA, 0-9 μM). Growth regulators (BA and NOA) in the culture medium were essential for induction of somatic embryogenesis since explants incubated on hormone-free medium never regenerated somatic embryos. Usually, the regenerated somatic embryos become visible as small white globular structures on the surface of the callus 3-4 months after culture initiation. The best embryogenic response (27 %) was obtained when stigma and style explants were cultured on NN medium supplemented with 5 μM NOA and 9 μM BA. Somatic embryos developed into plantlets when transferred to a hormone-free semisolid NN medium. 35 % of primary somatic embryos showed secondary embryogenesis

Influence of Culture Conditions on In Vitro Asymbiotic Germination of Anacamptis longicornu and Ophrys panormitana (Orchidaceae)

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

Leaf water relation traits in typical Sicilian varieties of Vitis vinifera L.

In Italy, grapevines are extensively cultivated, with Sicily representing one of the most significant wine regions. The high number of autochthonous grapevine varieties represents an important source of genetic diversity, and the many Sicilian varieties have anatomical and physiological traits that allow them to resist to different levels of drought stress. We investigated the water relation parameters of four cultivars of Vitis vinifera L. (Catarratto, Corinto, Nero d'Avola and Zibibbo) and characterized their leaf hydraulics. Measurements were conducted during summer on plants growing in the experimental field of the IBBR-CNR near Palermo. Daily patterns of leaf water potential (Yleaf) and stomatal conductance (gs) were measured in the field. Pressure-volume curves were constructed by the bench dehydration method to obtain leaf water potential at turgor loss point (Ytlp), osmotic potential at full rehydration (p0) and bulk modulus of elasticity (εmax). Leaf samples were collected to determine vein density using ImageJ. Major vein density was measured on digitally scanned leaves, while minor vein density was measured on photomicrographs of cleared and stained leaf portions

Urgent need for preservation of grapevine (Vitis vinifera L. subsp. vinifera) germplasm from small circum-Sicilian islands as revealed by SSR markers and traditional use investigations

Since the last decades grapevine germplasm is undergoing a process of rapid genetic erosion. This process is of particular concern in minor circum-Sicilian islands, because of the sharp reduction of the cultivated surfaces and the shift of their economy from agriculture to tourism. Aiming at valorising and preserving the surviving varieties we collected 185 accessions during several surveys since 2007. Six nuclear microsatellite markers were used for germplasm characterization, yielding 75 different genetic profiles. We found out that most genetic profiles (39) were not listed in national and international grapevine databases, confirming that the Sicilian minor islands represent underexplored hotspots of genetic diversity for grapevine. We also identified several synonymies, often due to geographic isolation, having 20 varieties at least two names. Conversely, 18 homonyms collectively indicated 34 genetically different accessions. Interviews with farmers provided information on current and past usage, and the origin and type of cultivation practices as well. The study also shows the urgent need for preservation of local grapevine germplasm, due to the disappearance of the elder caretakers of these traditional varieties. For rare germplasm preservation most part of the collected grapevine cultivars were introduced in an ex situ collection field

Regeneration of Algerian

Stigma/style somatic embryogenesis is one of the efficient methods in plant regeneration of most Citrus ssp., without inducing somaclonal variations. Furthermore, somatic embryogenesis from style/stigma proved to be effective in the elimination of the main citrus virus and virus-like diseases. This technique was applied on Algerian citrus collection. Different Citrus species [Citrus sinensis (L.) Osbeck, C. limon (L.) Burm, C. reticulata Blanco, C. paradisi Macfad, C. reshni Hort. ex Tan., C. jambhiri Lush and C. maxima (Burm.) Merrill] were chosen and tested for the presence of the main virus and virus-like agents. Most of the genotypes showed to be infected, mainly by viroid agents. Closed flowers were collected and in vitro cultured on a Murashige and Skoog (MS) medium supplemented with 6- benzylaminopurine. All explants produced callus about 4 to 9 days after culture initiation, whereas embryogenesis occurred after 38 to 150 days in most of the cultured genotypes. Formed embryos were cultured in a single tube before in vivo acclimatization. After sanitary assays, regenerated plants were shown to be free from the agents detected in the mother trees.Key words: Algeria, citrus germplasm, plant regeneration, sanitation, somatic embryogenesis

Autotetraploid Emergence via Somatic Embryogenesis in Vitis vinifera Induces Marked Morphological Changes in Shoots, Mature Leaves, and Stomata

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