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A simplification of a haemolytic micromethod is presented. In the orginal method, alfalfa (Medicago media Pers.) leaf sap is spotted on the plate covered with a blood-gelatine suspension. In the presented method, the mashed alfalfa pulp is used instead of sap. Due to saponin diffusion and the reaction with erythrocytes, a haemolytic ring appears, which has a width proportional to the concentration of toxic saponins. It is shown that the width of haemolytic ring does not depend on the sample weight ranging from 20 to 100 mg. This allows for the omission of laborious sap pressing and sample quantification. Individual alfalfa plants with different saponin contents were tested using leaf sap and leaf pulp for analyses. Good agreement was found with sap and leaf pulp methods. The correlation obtained by both methods was high, r = 0.87. The modified method requires only a small amount of plant material and makes the analyses of large numbers of individual plants per day possible. The method is especially recommended for breeding purposes.
The aim of this study was to determine the composition of nitrogen fractions in alfalfa and red clover, which differ in proteolytic activity, and to evaluate the effect of wilting on changes in nitrogen fractions in alfalfa and red clover herbage. Total nitrogen was divided into protein and non-protein nitrogen, and the amino acid profile of protein was analyzed. Buffer-soluble nitrogen (BSN), including buffer-soluble protein nitrogen (BSPN) and non-protein buffer-soluble nitrogen (NPBSN), was determined. The NPBSN fraction was further subdivided into peptide nitrogen, amino acid nitrogen, neutral detergent-insoluble nitrogen (NDIN) and acid detergent-insoluble nitrogen (ADIN). Wilting in the field to 40% dry matter content (the swath was tedded once) did not reduce the total nitrogen content of alfalfa and red clover herbage, although it affected the concentration of the BSNfraction, in particular NPBSN, and free amino acid nitrogen. During alfalfa wilting, the soluble protein content decreased and the concentrations of non-protein nitrogen compounds increased, mostly due to an increase in free amino acid nitrogen. A reverse trend was observed during red clover wilting – the concentrations of non-protein nitrogen compounds decreased and soluble protein content increased. A decrease was also noted in peptide nitrogen, NDIN and ADIN. Wilting of alfalfa and red clover had no adverse effect on the amino acid profile of protein. The concentrations of essential amino acids that limit milk protein synthesis, i.e. Lys, Met, His and Arg, did not decrease. Legume wilting in the field contributes to an increase in the concentrations of soluble nitrogen in the plant material intended for ensiling.
Bacterial wilt caused by Clavibacter michiganensis subsp. insidiosus (Cmi) is an integral part of chronic vascular wilt of alfalfa. The aim of the presented study was to determine the virulence of Cmi strains and later on to evaluate the level of resistance of alfalfa varieties registered in the Czech Republic and of some standard check varieties. Three methods of inoculation were compared at greenhouse conditions. The severity of the disease was determined on the basis of symptoms appearing on the cross section of the roots. The Cmi strain 12/5/98 and its re-isolates showed a high level of virulence. Freeze-dried isolates lost their virulence. The best differentiation of tested varieties according to the level of resistance was obtained by using successive inoculation on cotyledons, stems and roots. Of tested alfalfa varieties, Vernal and Roamer were classified as resistant and Nitranka, Orca and Europe as the most susceptible. Alfalfa varieties (Czech and European) registered in the Czech Republic recently proved lower disease severity in comparison with ones registered formerly (r = –0.538, p (0.05) = 0.553). The above mentioned fact evidences the progress of Cmi resistant alfalfa breeding in Europe
Poor seed set is a limiting factor in alfalfa breeding, as it slows the selection response. One strategy used to overcome this problem is to search for mutations of inflorescence morphology. Long-peduncle (lp), branched-raceme (br) and top-flowering (tf) inflorescence mutations increase the number of flowers per inflorescence, but they do not improve seed set per flower. Here we assessed pollen tube growth in styles of those inflorescence mutants and we observed embryo and endosperm development in seeds 1 to 16 days after pollination (DAP). The number of pollen tubes penetrating the style and the ovary was similar in all tested mutants and in the reference cultivar Radius. At 2 DAP, fertilized ovules were 2.7–3.9 times less numerous in certain inflorescence mutants than in the short-raceme cv. Radius. Ovule degeneration progressed at 2–4 DAP in all analyzed plants. Most ovules were not properly developed in the control cultivar (62%), nor in the forms with mutated inflorescence morphology (69–86%). The number of seeds per pod was lowest in the tf form despite its having the highest number of ovules per ovary. It appears that the number of ovules per pistil is not a crucial factor in seed set in alfalfa when fertilization efficiency is very low. Both poor fertilization and gradual ovule degeneration were factors causing poor seed set in the investigated alfalfa genotypes.
Value of beets (varieties: sugar beet AJ-Poly 1 and fodder beets: Poly Past, Albus and Cyklop) and lucerne forage or lucerne and timothy forage for production of combined silages for growing pigs was determined. The obtained silages were of very good quality and relatively high content of dry matter (about 40%). The balance studies indicated that there was no significant difference in digestibility of nutrients and in nitrogen balance related to variety of beets as well as to lucerne forage. Slightly better results were obtained in case of using of lucerne and timothy and also Poly Past beets. However, in terms of nutrients crop per 1 ha, the variety Albus (12362 oat units ) was the most efficient - except of sugar beets.
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The inspiration to reassess method and timing for establishing alfalfa (Medicago sativa L.) is meteorological data documenting that current Polish winters are milder than several decades ago, when management recommendations were worked out for this crop. Additionally, prolonged periods of spring drought occur more frequently, posing significant challenges to the development of alfalfa seedlings under a companion crop. The hypothesis was made that as climatic conditions change, the traditional practice of spring sowing with a barley (Hordeum vulgare L.) companion crop may become higher risk, and direct sowing later in the season when rainfall is more dependable would provide advantages. From 2010 to 2014 three series of experiments were carried out over the establishment year and first production year. Establishment success and yield performance of alfalfa sown with a barley companion crop in April was compared to alfalfa direct sown in May, June, July and August. Plant density, overwinter survival, and diameter of the upper taproot from sowings performed in the period April to late July were similar. Plant density with August sowing was significantly higher, but taproot diameter was smaller than in plants sown earlier in spring or summer. Plants from all sowing treatments overwintered well, with the greatest stand loss of only 15% occurring with August sowing. In the first production year, mean dry matter yields were similar for alfalfa sown in the period from April to late July, averaging 11.7 Mg. ha⁻¹. First production year yield associated with August sowing was significantly lower, at 8.88 Mg. ha⁻¹. In the years and locations of this study, the alternative of direct sowing alfalfa up to late July resulted in similar establishment success to the traditional practice of sowing with a barley companion crop in early spring.
The authors investigated the impact of the Sinorhizobium inoculum density on the plant development of alfalfa, nodulation and nitrogenase activity. It was found that plants inoculated with a 10% inoculant (4.9 x 10⁶ CFU) were characterized by the best growth, more profuse fresh material and a very well developed root system and, additionally, they revealed higher nitrogenase activity.
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