Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.     

Use of molecular markers to accelerate the breeding of common bean lines resistant to rust and anthracnose

The main goal of this work was to introduce resistance genes for rust, caused by Uromyces appendiculatus, and anthracnose, caused by Colletotrichum lindemuthianum, in an adapted common bean cultivar through marker-assisted backcrossing. DNA fingerprinting was used to select plants genetically closer to the recurrent parent which were also resistant to rust and to race 89 of C. lindemuthianum. DNA samples extracted from the resistant parent (cv. Ouro Negro), the recurrent parent (cv. Rudá), and from BC₁, BC₂ and BC₃ resistant plants were amplified by the RAPD technique. The relative genetic distances in relation to the recurrent parent varied between 9 and 59% for BC₁, 7 and 33% for BC₂, and 0 and 7% for BC₃ resistant plants. After only three backcrosses, five lines resistant to rust and anthracnose with, approximately, 0% genetic distance in relation to the recurrent parent were obtained. These lines underwent field yield tests in two consecutive growing seasons and three of them presented a good yield performance, surpassing in that sense their parents and most of the reference cultivars tested.     

铜绿微囊藻对苯酚的富集与降解

以苯酚为培养介质中的唯一碳源,研究铜绿微囊藻的生长和对苯酚的去除效果。结果表明,铜绿微囊藻能利用苯酚的碳进行光合作用,加快其生长;同时,铜绿微囊藻对苯酚有明显的富集与降解作用,富集量２４ｈ最大达２７．８５ｍｇ／ｇ,此后随时间的延长逐渐降低。铜绿微囊藻对苯酚的降解符合一级动力学方程,５ｄ降解率为６７．６％。     

Relationships among American and Spanish populations of maize

Summary Two experiments were carried out with two objectives. First, to establish the phenetic relationships among the maize (Zea mays L.) landraces from Galicia (Northwestern Spain) maintained at the Misión Biológica de Galicia. Second, to assess the resemblance between a collection of Spanish populations (including the landraces from Galicia) and a set of US Corn Belt varieties. For the first objective 73 varieties from Galicia, along with 9 hybrid checks, were grown in 9×9 simple lattices at two locations for two years. For the second objective 131 populations from the US Corn Belt and Spain, along with 9 hybrid checks, were grown for three years in unreplicated experiments. Cluster analyses were carried out with the first principal components that accounted for a significant amount of the total variation. Four groups were found among the landraces from Galicia. The populations from Spain and America were classified as belonging to nine main groups. The replicated experiment was more accurate than the unreplicated one. However, it is concluded that an unreplicated test grown in several environments is accurate enough to detect the main groups, although some inaccuracies should be expected.     

Damage Constitutive Equation and Experiment of Layered Rock

The microcosmical study on many speciments of sedimentary rock by SEM and petrography theory show that rock is formed of microcrystalline and micropore with all kinds of forms.Therefrom the microstructure model of rock is established.Further the three dimensional nonlinear damage constitutive equation and damage evolution equation of layered rock including respective process of damage evolution,mechanical parameter,effective stresses,and unequal biaxial stress in different rocks are derived.The results of the experiment indicate that the theoretical curve is fairly close to the experimental curve.     

Chromosome localisation of genes for resistance to Heterodera schachtii, Cercospora beticola and Polymyxa betae using sets of Beta procumbens and B. patellaris derived monosomic additions in B. vulgaris

Beet cyst nematodes (BCN, Heterodera schachtii), Cercospora beticola, and rhizomania, caused by the beet necrotic yellow vein virus (BNYVV) and vectored by the soil-borne fungus Polymyxa betae, are the most serious diseases of sugar beet ( Beta vulgaris subsp. vulgaris). The wild Beta species of section Procumbentes are known to be completely resistant to H. schachtii, C. beticola and P. betae. Alien monosomic additions (2n=19), plants of cultivated beet (2n=18) carrying different individual chromosomes of B. procumbens (2n=18) or B. patellaris (2n=36), were tested in greenhouse experiments for resistance to these pathogens. Gene(s) conferring full resistance to the beet cyst nematode in B. patellaris are located on chromosome 1.1, and the other tested chromosomes of B. patellaris are not involved in the expression of resistance. Artificial inoculation under greenhouse conditions, with in vitro produced inoculum of C. beticola and spot-percentage rating of the disease intensity, showed that the high level of resistance that was observed in the wild species B. procumbens and B. patellaris was not found in any of the monosomic additions tested. It was suggested that genes on various chromosomes of the wild species are needed to express full resistance, and that the chromosomes of group 7 of B. patellaris and chromosome 7 of B. procumbens have the largest effect. The greenhouse tests for resistance to P. betae in B. patellaris derived monosomic additions showed that the addition families of group 4.1 have a strong partial resistance, while the addition families of group 8.1 appeared to be completely resistant to the pathogen. Resistance to P. betae in the two wild species as well as in the two resistant addition types did not exclude infection with BNYVV, but resulted in a considerable reduction of the virus concentration. It was concluded that resistance to the vector would complement virus resistance, and may provide a more effective and durable control of rhizomania. This revised version was published online in July 2006 with corrections to the Cover Date.