Inheritance of resistance to the papaya ringspot virus-watermelon strain from two different accessions of winter squash Cucurbita maxima Duch.
Inheritance of resistance to the papaya ringspot virus-watermelon strain (= PRSV-W, syn. watermelon mosaic virus-1 = WMV-1) was studied for two Cucurbita maxima accessions – ABL-010 and Redlands Trailblazer – considered to be resistant to the potyvirus. ABL-010 (ABL) derives its resistance from Brazilian accessions of C. maxima. Redlands Trailblazer (RT) is an Australian cultivar of C. maxima, the resistance of which traces back to C. ecuadorensis. In both cases, virus could be recovered from the inoculated (symptomless) plants via back inoculation onto susceptible C. pepo cultivar Caserta, indicating a tolerance type of resistance. Both ABL and RT were crossed to each other and to the standard PRSV-W-susceptible cultivar Buttercup (B). For each one of the crosses (ABL ×> B), (ABL × RT) and (RT × B), parental lines, F1, F2 and backcrosses to both the parents (BC1P1 and BC1P2) were tested for PRSV-W resistance after mechanical inoculation. Seedlings were inoculated at the first true leaf stage with a known PRSV-W isolate in 0.02 M phosphate buffer (pH 7.0) plus sodium sulphite 0.01%. Plants were scored for virus symptoms 28 days after inoculation; symptom severity being rated with a scale from 0 (= no symptoms) to 5 (= plants with severe mosaic and leaf distortions). Broad and narrow-sense heritabilities of PRSV-W resistance were 0.44/0.36, 0.59/0.59 and 0.58/0.58 for crosses (ABL × B), (ABL × RT) and (RT × B), respectively. Resistance imparted by ABL in cross with susceptible B could be explained by the action of three genes with partial dominance. Resistance imparted by RT in cross with B was found to be under control of at least two genes with additive effects. Susceptible plants were found in segregating populations of the cross ABL × RT (transgressive segregation), indicating that at least one of the loci involved in the control of resistance in ABL was not allelic to their counterpart loci in RT. The resistance imparted by RT appears to be of higher level than that imparted by ABL.
Sodium sulphite inhibition of potato and cherry polyphenolics in nucleic acid extraction for virus detection by RT-PCR.
Phenolic compounds from plant tissues inhibit reverse transcription-polymerase chain reaction (RT-PCR). Multiple-step protocols using several additives to inhibit polyphenolic compounds during nucleic acid extraction are common, but time consuming and laborious. The current research highlights that the inclusion of 0.65 to 0.70% of sodium sulphite in the extraction buffer minimizes the pigmentation of nucleic acid extracts and improves the RT-PCR detection of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in potato (Solanum tuberosum) tubers and Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV) in leaves and bark in the sweet cherry (Prunus avium) tree. Substituting sodium sulphite in the nucleic acid extraction buffer eliminated the use of proteinase K during extraction. Reagents phosphate buffered saline (PBS)-Tween 20 and polyvinylpyrrolidone (PVP) were also no longer required during RT or PCR phase. The resultant nucleic acid extracts were suitable for both duplex and multiplex RT-PCR. This simple and less expensive nucleic acid extraction protocol has proved very effective for potato cv. Russet Norkotah, which contains a high amount of polyphenolics. Comparing commercially available RNA extraction kits (Catrimox and RNeasy), the sodium sulphite based extraction protocol yielded two to three times higher amounts of RNA, while maintaining comparable virus detection by RT-PCR. The sodium sulphite based extraction protocol was equally effective in potato tubers, and in leaves and bark from the cherry tree.
Reference values for some haematological, haematochemical, and electrophoretic parameters in the Girgentana goat
The purpose of this study was to determine haematolog ical, haematochemical, and electrophoretic reference values for the Girgentana goat in order to form a basis for clinical interpretation. The study included 348 female Girgentana goats aged from 1 to 6 years. The animals were divided into 3 groups: goats aged 1-2 years (group A), goats aged 3-4 years (group B), and goats aged 5-6 years (group C). Blood samples were collected from each animal every morning at the same time (0800) by means of jugular venipuncture, and red blood cell (RBC), white blood cell (WBC), neutrophil (NEU), lymphocyte (LYM), monocyte (MONO), eosinophil (EOS), and basophil (BASO) counts, and haemoglobin (Hb), packed cell volume percent (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and red cell distribution width (RDW) were recorded. Additionally, the concentration of some haematochemical parameters, including alanine-aminotransferase (ALT), aspartate-aminotransferase (AST), γ-glutamine-transferase (γ-GT), creatinine, β-hydroxybutyrate, urea, non- esterified fatty acids (NEFA), potassium, sodium, and chloride, and the electrophoretic profile (albumin, α1-globulins, α2-globulins, β-globulins, γ-globulins, total proteins and albumin:globulins ratio) were determined. Statistical analysis (one-way analysis of variance (ANOVA)), followed by Bonferroni's test, showed that there were significant differences only in RBC, MCV, EOS, ALT, AST, creatinine, γ-GT, sodium, urea, NEFA, γ-globulins, and the albumin:globulins ratio (P < 0.05 was considered statistically significant). The values determined in the present study contribute to the knowledge of the reference ranges for the Girgentana goat and can be used for monitoring health and disease diagnosis.