A set of primers for analyzing chloroplast DNA diversity in Citrus and related genera

Chloroplast simple sequence repeat (cpSSR) markers in Citrus were developed and used to analyze chloroplast diversity of Citrus and closely related genera. Fourteen cpSSR primer pairs from the chloroplast genomes of tobacco (Nicotiana tabacum L.) and Arabidopsis were found useful for analyzing the Citrus chloroplast genome (cpDNA) and recoded with the prefix SPCC (SSR Primers for Citrus Chloroplast). Eleven of the 14 primer pairs revealed some degree of polymorphism among 34 genotypes of Citrus, Fortunella, Poncirus and some of their hybrids, with polymorphism information content (PIC) values ranging from 0.057 to 0.732, and 18 haplotypes were identified. The cpSSR data were analyzed with NTSYS-pc software, and the genetic relationships suggested by the unweighted pair group method based on arithmetic means (UPGMA) dendrogram were congruent with previous taxonomic investigations: the results showed that all samples fell into seven major clusters, i.e., Citrus medica L., Poncirus, Fortunella, C. ichangensis Blanco, C. reticulata Swingle, C. aurantifolia (Christm.) Swingle and C. grandis (L.) Osbeck. The results of previous studies combined with our cpSSR analyses revealed that: (1) Calamondin (C. madurensis Swingle) is the result of hybridization between kumquat (Fortunella) and mandarin (C. reticulata), where kumquat acted as the female parent; (2) Ichang papeda (C. ichangensis) has a unique taxonomic status; and (3) although Bendiguangju mandarin (C. reticulata) and Satsuma mandarin (C. reticulata) are similar in fruit shape and leaf morphology, they have different maternal parents. Bendiguangju mandarin has the same cytoplasm as sweet orange (C. sinensis), whereas Satsuma mandarin has the cytoplasm of C. reticulata. Seventeen PCR products from SPCC1 and 21 from SPCC11 were cloned and sequenced. The results revealed that mononucleotide repeats as well as insertions and deletions of small segments of DNA were associated with SPCC1 polymorphism, whereas polymorphism generated by SPCC11 was essentially due to the variation in length of the mononucleotide repeats.     

Genetic diversity in Tunisian olive accessions and their relatedness with other Mediterranean olive genotypes

Eighty-four olive accessions obtained from the National Conservatory of Boughrara-Sfax (Tunisia), previously evaluated for morphological traits, were analysed with 47 random amplified polymorphic DNA (RAPD) markers. They were compared with other olive genotypes originated from Eastern or Western Mediterranean. The highest and lowest similarities between genotypes, estimated by simple matching algorithm, were 0.98 and 0.40, respectively. A dendrogram based on Ward's method and a factorial correspondence analysis (FCA) showed that most of Tunisian accessions are closely related to olive genotypes originating from the Eastern Mediterranean and some are clustering with genotypes originated from the Western Mediterranean. These findings suggested multiple and complex origin of Tunisian olive. A comparative study between a previous morphological analysis and current RAPD assay was carried out and discussed.     

Temperature and storage period over spermatic parameters of jundiá, Rhamdia quelen (Quoy & Gaimard, 1824)

The aim of this work was to evaluate the spermatic parameters of jundia, Rhamdia quelen, semen stored for short periods under different temperatures. Fifteen males were used in a time factorial experimental design (13 × 3 × 3 × 3). The sperm was stored at: 15; 25 and 35°C and activated in water at: 15; 25 and 35°C each, respectively, within storage periods of: 0; 2; 4; 6; 8; 10; 12; 16; 20; 24; 32; 40 and 48 h after collection. The treatments were performed in triplicates and in sequential protocols every 50 h. The motility parameters were evaluated using Pearson's correlation analysis and the significant parameters (P < 0.05) were summarized using principal component analysis forming two predicted groups. Group 01 composed the following parameters: curvilinear velocity, wobble and linearity; group 02 comprised average path velocity, straight line velocity, beating cross frequency and motility rate. Effects (P < 0.05) of treatments were observed only in group 02, with interaction between time and temperature of exposure and between exposure time and water temperature. Superior results of spermatic parameters of group 02 were observed right after collection. However, the exposure and spermatic activation at 15°C assured significant results 48 h after collection when compared with 25 and 35°C.     

Masculinization of the Native Bay Snook,Petenia splendida,Using Oral Administration of the Synthetic Steroid 17α‐methyltestosterone

The possibility of inducing sex reversal in bay snook, Petenia splendida, was evaluated to generate a protocol aimed at producing single‐sex populations (100% males) and improving farming conditions for this species. This popular staple food is a cichlid fish native to Central America, with aquaculture potential. We evaluated oral administration of the synthetic steroid 17α‐methyltestosterone (MT) to determine optimal dosage (30, 45, and 60 mg/kg of food) and days of feeding (30, 45, and 60) to attain efficient masculinization. The statistical analysis indicates a highly significant effect of both factors as well as for the interaction (P < 0.01). We registered significant differences in growth at the end of the experiment; fish fed with 60 mg of MT for 60 d reached the largest size, while fish fed with no MT were smaller. No significant differences were observed for survival (P > 0.05), ranging from 55.7 to 75.3% in the treatments. We recommend the use of 60 mg/kg of MT for 30 d for achieving successful sex reversal. This study provides valuable information supporting the inclusion of new species in aquaculture, reducing time and costs associated with sex reversal previously proposed for the species.