ABSTRACT
Rocket is the common designation for two baby-leaf salad crops of the Brassicaceae family: Eruca sativa (L.) Cav., usually referred to as annual garden rocket, and Diplotaxis tenuifolia (L.) DC. commonly named to as perennial wild rocket. E. sativa is used for human consumption since antiquity. However, the growing consumer preference for D. tenuifolia is being accompanied by the fast increase in its production area and commercialization of new cultivars. Nevertheless, the worldwide number of wild rocket accessions maintained in germplasm collections is very reduced, the solution for which situation the project "REMIRucula" intends to contribute, establishing a germplasm collection at the INIAV, Oeiras, Portugal. Herein, we report on the establishment via next generation sequencing (NGS) of the first genome assembly of D. tenuifolia and the identification of specific single sequence repeat (SSR) and single nucleotide polymorphisms (SNP) loci for the establishment of specific DNA-markers for this species. A representative set of 87 D. tenuifolia and 3 E. sativa accessions were assessed by 5 SSR and 9 SNP-CAPS markers, allowing a drastic discrimination between both species and the establishment of unequivocal molecular fingerprints for the analyzed accessions. The non-discrimination within six pairs and one trio of D. tenuifolia accessions is discussed.
ABSTRACT
The technique of laparoscopic oocyte aspiration has been increasingly used in animals; however, there are few records of its use in buffaloes. To describe this technique, six suckling Murrah buffaloes aged between 3 and 5 months were used. Three laparoscopic ovum pick-ups were performed in each animal, with intervals of 15 days between surgeries, completing a total of 18 procedures. The technique used three surgical ports with optics and a high-definition video camera. The introduction of the first portal and insufflation of the abdomen was performed through the open technique, with aspiration using a 20 G needle transabdominally and a vacuum pump calibrated at 50 mmHg. The mean complete surgical time from anesthesia to the removal of the animal from the litter was 49 ± 9.8 min. There were 27.8% cases of insufflation on the wrong side of the omentum. The oocyte recovery rate of 60.3% remained within the normal range. However, the rate of viable oocytes recovered was low, with only 40.8% of those recovered undergoing in vitro embryo production (IVEP). These data demonstrate that this simple, minimally invasive technique is an excellent reproductive tool for the genetic improvement of buffalo species.
ABSTRACT
We describe the construction of a BAC contig and identification of a minimal tiling path that encompass the dominant and monogenically inherited downy mildew resistance locus Pp523 of Brassica oleracea L. The selection of BAC clones for construction of the physical map was carried out by screening gridded BAC libraries with DNA overgo probes derived from both genetically mapped DNA markers flanking the locus of interest and BAC-end sequences that align to Arabidopsis thaliana sequences within the previously identified syntenic region. The selected BAC clones consistently mapped to three different genomic regions of B. oleracea. Although 83 BAC clones were accurately mapped within a â¼4.6 cM region surrounding the downy mildew resistance locus Pp523, a subset of 33 BAC clones mapped to another region on chromosome C8 that was â¼60 cM away from the resistance gene, and a subset of 63 BAC clones mapped to chromosome C5. These results reflect the triplication of the Brassica genomes since their divergence from a common ancestor shared with A. thaliana, and they are consonant with recent analyses of the C genome of Brassica napus. The assembly of a minimal tiling path constituted by 13 (BoT01) BAC clones that span the Pp523 locus sets the stage for map-based cloning of this resistance gene.
