Production and characterization of new triploid seedless progenies for mandarin improvement

Eighty-six triploid Citrus plants were recovered from interploid crosses between a natural tetraploid selection of the tangerine ‘Dancy’ (Citrus reticulata Blanco, cultigroup ‘Tangerine’), used as the pollen parent, and two seedy selections of diploid mandarins (C. reticulata Blanco) as well as one clementine (Citrus clementina Hort. ex Tan.): ‘Fortune’ mandarin, ‘Wilking’ mandarin and ‘Monreal’ clementine. Flow cytometric analysis was used for screening the triploid plantlets and the ISSR-PCR technique was used to characterize the obtained triploids through a double approach, confirming the hybrid nature of the offspring and allowing the analysis of the genetic pool obtained. Selection among triploid genotypes generated from elite seedy parents will give us a better chance to obtain superior mandarin cultivars characterized by true seedlessness.     

Monoclonal Antibody Capture Fluorometric Enzyme-Linked Immunosorbent Assay for Detection of Equine Growth Hormone in Plasma

Veterinary Research Communications -     

Effects of the Gpc‐B1 locus on high grain protein content introgressed into Argentinean wheat germplasm

Wheat grain protein content (GPC) is important for human nutrition and has a strong influence on the quality of pasta and bread. The objective of this study was to analyse the introduction of the Gpc‐B1 allele into two Argentinean bread wheat cultivars. Near‐isogenic lines were developed in ‘ProINTA Oasis’ and ‘ProINTA Granar’ using marker‐assisted selection. Gpc‐B1 lines showed a significant (P = 0.01) increase in GPC and a significant (P = 0.001) decrease in grain weight in comparison with control lines without Gpc‐B1. Differences in yield were not significant (P = 0.49) between lines. Gpc‐B1 lines significantly reduced (P = 0.02) straw nitrogen concentration at maturity and significantly increased (P = 0.02) the nitrogen harvest index. When data were analysed by genotype and environment, differences in some analysed parameters were found, indicating that Gpc‐B1 expression may be affected by different genetic backgrounds and environmental conditions. These results suggest that the introgression of the Gpc‐B1 allele into Argentinean wheat germplasm could be a valuable resource for improving GPC with no detrimental effect on grain yield.     

Morphology of the Lingual Dorsal Surface and Oral Taste Buds in Italian Lizard (Podarcis sicula)

With 4 figures The Italian lizard (Podarcis sicula) is the most diffused reptile in Italy, but it is also present in other European countries. This lizard belongs to the Lacertidae family, lives near walls, slants and along the borders of the paths; its diet includes bugs and aracnids. No data are so far available in literature about the three‐dimensional morphology of the tongue of Podarcis sicula, therefore the aim of the present paper was to study by scanning electron and light microscopy the three‐dimensional characteristics of the dorsal lingual surface and moreover the presence of chemosensory receptors like the taste buds in the oral cavity. Our results demonstrate that the Podarcis sicula tongue is a triangular muscular membranous organ, dorsoventrally flattened and that three different areas can be observed: a bifid apex, a body and a root. No papillae were observed in the apex, characterized by a flattened mucosa and by two deep median pouches. In the body cylindrical papillae with a flat surface are present, aborally gradually substituted by imbricated papillae. Foliate‐like papillae were observed in the lateral parts of the tongue body. No sensory structures were showed on the lingual dorsal surface, while they were numerous in the oral cavity, particularly on the gingival epithelium. The light microscopy shows, on the dorsal surface, a stratified pavimentous not keratinized epithelium, conversely keratinized along the ventral surface. Many caliciform cells on the lateral parts of the papillae, deputed to the secretion of mucus, were also observed. Therefore, the results obtained in this paper could give a contribution to the knowledge of the tongue anatomy in a species widely diffused in different European countries and could be of help for clinical purposes in reptiles.     

Assessment of the origin of new citrus tetraploid hybrids (2<Emphasis Type="Italic">n</Emphasis> = 4x) by means of SSR markers and PCR based dosage effects

We report the accurate determination of the allelic configurations of a total of eight new citrus tetraploid hybrids by means of SSR analysis, coupled with capillary electrophoresis, and PCR based dosage effects. Tetraploid hybrids were spontaneously obtained from different interploid crosses (2x × 4x) between diploid ‘Femminello’ lemon and the allotetraploid somatic hybrid (2n = 4x = 36) ‘Key’ lime + ‘Valencia’ orange, and between diploid ‘Wilking’ and ‘Fortune’ mandarins and an autotetraploid ‘Dancy’ mandarin (2n = 4x = 36). To understand the opportunity to employ them in further backcross programs, the cytological mechanisms underlying their ploidy level were unambiguously determined using six SSR primers. PCR conditions were optimized and skewness in template/product ratios were verified. Tetraploid allelic configurations were determined from PCR based dosage effects using electropherogram peak heights to estimate the copy number per allele. In all the tetraploid hybrids we found out that diploginy (2n eggs) has occurred, contributing the extra haploid genome in the tetraploids. According to the marker genotypes, it was further inferred that the 2n eggs in ‘Femminello’ lemon resulted from first division restitution (FDR), while in ‘Wilking’ and ‘Fortune’ mandarins 2n eggs occurred in second division restitution (SDR). These new genotypes, with their improved genetic female background, can be therefore considered very valuable in our citrus genetic improvement program as pollen donors in backcrosses suitable to eliminate negative traits.     

The Lingual Dorsal Surface of the Blue-Tongue Skink (Tiliqua scincoides)

The blue-tongue lizard ( Tiliqua scincoides ) is a variety of large skink common throughout Australia. There are seven species of Tiliqua and all of them have long bodies, short limbs and short and robust tails. T. scincoides occurs in a wide range of habitats; its diet is omnivorous. When threatened, it opens the mouth and protrudes its characteristic large fleshy cobalt blue tongue. It is currently found as a popular species and also as a pet animal in the European countries. No data are available in literature about the morphology of the tongue of T. scincoides ; therefore, the aim of the present study was to investigate by means of scanning electron microscopy and light microscopy, the anatomy of the dorsal lingual surface. Our results demonstrate the presence of a tongue tip with a smooth surface without papillae. The foretongue was characterized by a stratified epithelium with foliate-like papillae and deep inter-papillar spaces in the middle part and cylindrical papillae with a flat surface in the lateral parts. All the posterior area of the tongue was characterized by more compacted papillae and the inter-papillar spaces were very narrow. Light microscopy showed the presence of melanin throughout the tongue. No taste buds were recognized on the lingual dorsal surface. Therefore, the papillae probably have a mechanical function showing an important role in the swallowing phase. The morphology of the tongue surface can be correlated to the diet and, different roles, as in other examined species, can be hypothesized for different areas.     

The tongue of Leopard Gecko (Eublepharis macularius): LM,SEM and confocal laser study

The leopard gecko is a crepuscular and insectivorous reptile. The role of the tongue in this reptile is fundamental for the prey capture and ingestion and is not related with eyes cleaning as usual in other geckos. The elongated tongue can be divided into a foretongue with a slightly bifurcated apex and a hindtongue. Scanning electron microscopy demonstrated that several different papillae are present on the dorsal surface, foliate and dome-shaped in the foretongue, becoming thicker and stouter with reduced interpapillary spaces in the lateral parts. The hindtongue is characterised by wide foliate papillae with indented margins and deep fissures of the mucosa. Light microscopy showed the presence of a stratified slightly keratinized squamous epithelium in the apex of the foretongue, a stratified non-keratinized squamous epithelium in the fore and in the hindtongue. In the foretongue, numerous muciparous caliciform cells were observed. Moreover, the presence of taste buds on the tongue ventral surface was demonstrated for the first time in this species and the confocal laser study revealed a strong immunoreactivity for the S-100 protein in the sensory cells. Therefore, the results obtained could give a contribution to the knowledge of the tongue anatomy and are a basis for eventual further studies regarding the feeding habits in a reptile become a popular pet.     

The Tongue Dorsal Surface in Fish: A Comparison Among Three Farmed Species

The oral cavity of vertebrates has a very important role in many functions strictly related to the food processing. Two species, sea bass Dicentrarchus labrax and seabream Sparus aurata, are mainly involved in the aquaculture industry in the mediterranean area; moreover, the white seabream Diplodus sargus sargus was recently chosen as the best candidate for the diversification of species of commercial interest. This investigation was carried out, using the standard procedures for light and scanning electron microscopy, to analyse the morphology of the tongue dorsal surface in order to show whether relationships are present between the tongue morphology and the nutritional choices of these species. In all the three investigated species, three different areas were observed on the dorsal lingual surface: an apex, a body and a root. In the sea bass D. labrax, numerous caninelike teeth, organized in pads, were present along the dorsal surface. The presence of numerous taste buds, scattered on the tongue, was shown. In the seabream S. aurata, the apical part of the tongue is inserted in a pouch with a characteristic medial ridge on the body surface. Numerous taste buds were scattered on the whole surface. In the white seabream D. sargus sargus, a pouch partially covering the apex was clearly observed. In the pouch, filiform‐like and foliate‐like papillae can be distinguished. The results show the important role of the fish tongue in the food ingestion process, demonstrating the interaction of food processing, taste and morphological characteristics.     

Orexin A Expression in the Ovary of Dog and Cat

Orexin A and B, also known as hypocretin A and B, are hypothalamic neuropeptides arising from a precursor to the 130 amino acid, called pre–pro orexin. They are synthesized mainly in lateral and posterior hypothalamus and are involved in different functions such as regulation of food intake and energy balance. Orexins and orexin receptors were previously described also in different tissues and organs outside the brain. The aim of this study was to demonstrate by means of the immunofluorescence technique, the presence of orexin A in the ovary of cat and dog, to support the hypothesis of the role of this substance also at the level of the female genital system. The presence of orexin A in the ovary either in dog or in cat is in agreement with previous data on the presence and role of orexins in the female genital system of other species.     

Testicular leiomyoma and spermatogenic failure syndrome in a seabass from broodstock

Testicular leiomyomas and gonadal dysfunctions are not commonly reported in teleost and particularly in broodstock fish. In the present work, a testicular leiomyoma related to an unusual case of spermatogenic failure syndrome, in a broodstock seabass coming from an Italian aquaculture farm, is reported. At gross pathology, the circumscribed neoplasia showed several white nodular masses, originating from the gonad, deforming the anatomo-physiological shape. From light microscopy, the presence of neoplastic tissue islands with spindle cells with a whirling pattern and the low presence of connective tissue were evident and originated from smooth muscle cells. Seminiferous tubules showed severe azoospermia with the absence of sperm cells even closer to the neoplastic area. Immunohistochemical analysis was positive for alpha-SMA, desmin and vimentin but negative for S-100, confirming a diagnosis of leiomyoma. This is the first report of spermatogenic failure syndrome associated with a testicular leiomyoma in fish from broodstock. More efforts should be made in studying broodstock fish pathologies related to fish maintenance and hormonal treatments that could economically affect aquaculture production.