Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Seed dormancy and germination in the summer annual Galeopsis speciosa

This study examined germination and dormancy in Galeopsis speciosa (Lamiaceae), a common summer annual weed in cold‐temperate areas. Seeds collected in southern Sweden were subjected to several experiments. The seeds were dormant at maturity. Seeds sown outdoors after collection produced a small number of seedlings that emerged early in the spring. After long cold stratification or stratification outdoors over two winters, the maximum germination was 40–50%; germination occurring over a wide range of temperatures. Warm stratification preceding cold stratification had no effect on germination, but repeated warm and cold periods seemed to promote germination. Gibberellic acid (GA) stimulated germination, but full germination was only achieved after more than 2 months of incubation at the most suitable temperature regime tested. Excised embryos grew and developed into normal seedlings. With these results, the species does not fit into the currently used system for seed dormancy classifications. The response to GA and the growth of excised embryos indicate non‐deep or intermediate physiological dormancy, but dormancy alleviation by stratification was not in line with the guiding principles for these classifications. Galeopsis speciosa has a strong dormancy that is sufficiently alleviated during the winter to allow germination of only part of a seed batch each year; hence a stepwise germination pattern occurs over a period of several years.     

Chemical Composition of Green Tea (<Emphasis Type="Italic">Camellia sinensis</Emphasis>) Infusions Commercialized in Portugal

To evaluate the potential benefits and risks associated with tea consumption it is important to identify the constituents of this beverage. Levels of some minerals, caffeine and catechins in green tea samples commercialized in Portugal were evaluated. Potassium is the metal present in larger amount (92–151 mg/l). The content of sodium, calcium, fluoride, aluminium, manganese and iron were 35–69, 1.9–3.5, 0.80–2.0, 1.0–2.2, 0.52–1.9, 0.020–0.128 mg/l, respectively. Chromium and selenium were not detected. The resulting data showed considerable variability in catechins content. The levels of epigallocatechin gallate (EGCG) ranged from 117 to 442 mg/l, epicatechin 3-gallate (EGC) from 203 to 471 mg/l, epigallocatechin (ECG) from 16.9 to 150 mg/l, epicatechin (EC) from 25 to 81 mg/l and catechin (C) from 9.03 to 115 mg/l. Caffeine contents in the green tea infusions studied were between 141–338 mg/l. Green tea infusions provide significant amounts of catechins and could be an important source of some minerals.     

Genetic characterization of banana cultivars (Musa spp.) from Brazil using microsatellite markers

Flavonoids, in particular the anthocyanins,are responsible for flower colour in manyspecies. The dihydroflavonols represent abranch point in flavonoid biosynthesis,being the intermediates for production ofboth the coloured anthocyanins, through theaction of the enzyme dihydroflavonol4-reductase (DFR), and the colourlessflavonols, produced by flavonol synthase(FLS). In this study the white-flowered,flavonol accumulating Mitchell line ofpetunia was used as a model to examine theinteraction between DFR and FLS enzymeactivities and possibilities forredirecting flavonoid biosynthesis awayfrom production of flavonols and towardsanthocyanins. Introduction of a 35SCaMV-DFR sense transgene construct causedthe production of anthocyanins, resultingin a pink-flowered phenotype. Furthermore,inhibition of FLS production throughintroduction of an FLS antisense RNAconstruct also led to anthocyaninproduction and a pink-flowered phenotype. A combination of both transgenes gave thehighest level of anthocyanin formation. Anthocyanins were produced in the DFR-senseand FLS-antisense transgenic lines in spiteof the greatly reduced levels of geneexpression in the Mitchell line for threeenzymes late in anthocyanin biosynthesis,anthocyanindin synthase, UDP-glucose:flavonoid 3-O-glucosyltransferase andUDP-rhamnose: anthocyanidin-3-glucosiderhamnosyltransferase. Thus, the level ofgene activity required for visibleanthocyanin formation is much lower thanthe high levels normally induced duringpetal development. Altering the balancebetween the DFR and FLS enzyme activities,using genetic modification, may be a usefulstrategy for introducing or increasinganthocyanin production in target ornamentalspecies.     

The impact of United States recreational fisheries on marine fish populations

We evaluated the commercial and recreational fishery landings over the past 22 years, first at the national level, then for populations of concern (those that are overfished or experiencing overfishing), and finally by region. Recreational landings in 2002 account for 4% of total marine fish landed in the United States. With large industrial fisheries excluded (e.g., menhaden and pollock), the recreational component rises to 10%. Among populations of concern, recreational landings in 2002 account for 23% of the total nationwide, rising to 38% in the South Atlantic and 64% in the Gulf of Mexico. Moreover, it affects many of the most-valued overfished species-including red drum, bocaccio, and red snapper-all of which are taken primarily in the recreational fishery.     

Genetic diversity of Musa diploid and triploid accessions from the Brazilian banana breeding program estimated by microsatellite markers

Banana (Musa spp.) is one of the most consumed fruits worldwide. Production is based mainly on triploid cultivars, and most genetic improvement programs aim to generate tetraploid hybrids obtained from the crossing of established triploid cultivars with a diploid parent genotype, improved or wild, exhibiting the trait of interest, normally resistance to biotic factors. Microsatellites were used to investigate the genetic variability and relationships between 58 Musa genotypes, including 49 diploids and nine triploid cultivars maintained at the Musa germplasm collection of the Brazilian dessert banana breeding program. Thirty-three primer pairs developed for banana were tested, and nine amplified reproducible and discrete fragments, producing a total of 115 alleles. The average number of alleles amplified per primer was 12.8, ranging from 10 to 15. The diploid genotypes presented the largest genetic variability, demonstrated by the large number of alleles detected, and the low similarity between the clones. The phenetic analysis clustered the triploid cultivars in a separated group, with the exception of the Nanica and Gros Michel cultivars, which showed high similarity with the diploid cultivar Mambee Thu. It was not possible to separate the wild diploid genotypes from the cultivated ones, indicating a common origin of these genotypes. A high proportion of duplicated alleles and/or loci was observed for diploid and triploid genotypes. The information gathered about the similarity between diploid and triploid accessions will help to define potential crosses to maximize the recovery of the typical fruit qualities required in Brazil (AAB, Pome and Silk dessert banana).