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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.
В работе описаны исследования коагуляции в зеленом соку из люцерны при помощи нефелометра. Исследовались процессы коагуляции при pH натуральном и при pH, доведенном до 9,4, в диапазоне температур 20-85°С. Исследовательский материал получено во время сжима проб люцерны под соответствующей нагрузкой. Для выяснения проблематики коагуляции использовано методы статической физики. Полученные результаты указывают на изменчивость исследуемых признаков в зависимости от разных факторов.
In the present work it was shown that total saponins originated from M. hybrida and M. sativa substantially limited mycelium growth of F. oxysporum f. sp. tulipae and symptoms of fusariosis on tulip bulbs. Out of 15 individual tested saponins originated from M. arabica, M. hybrida and M. sativa, four compounds: 3-O-[β-D-glucopyranosyl (1→2)α-L-arabinopyranosyl] hederagenin, hederagenin 3-O-β-D-glucopyranoside, medicagenic acid, medicagenic acid 3-O-β-D-glucopyranoside had the strongest inhibitory effect on mycelium growth of Fusarium oxysporum f. sp. tulipae on PDA medium. The total saponins from M. arabica, M. hybrida and M. sativa inhibited the number of colony forming units of Fusarium oxysporum f. sp. tulipae in artifi cially infested substrate. The use of saponins originated from Medicago as a fungicide is suggested.
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.
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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.
Flavonoids are a group of secondary metabolites found in most families. They are known to have important physiological functions in plants by protecting them against biotic stresses. Liquid chromatography (HPLC) was used to determine the flavonoid profiles, especially apigenin glycosides, their total concentration, as well as changes in the amount of six flavones found in the aerial parts of alfalfa (Medicago sativa L.) (Fabaceae) Radius cv. for three vegetative stages, uninfested and infested by the pea aphid (Acyrthosiphon pisum Harris) (Homoptera: Aphididae). It has been shown that both control and infested green aerial parts of alfalfa plants had similar flavonoid profiles. The dominant flavonoid of alfalfa was compound 7-O-[2-O-feruloyl- β-D-glucuronopyranosyl(1→2)-O-β-D-glucuronopyranosyl]-4’-O-β-D-glucuronopyranosideapigenin. Compound 4’-O-β-D-glucuronopyranosideapigenin was present in the smallest amounts. The total concentration of flavones was rather high and ranged from 10.32 to 12.28 mg/g d.m., but there were no significant differences between uninfested and infested alfalfa plants. There was a negative correlation between the concentration of total apigenin glycosides in the alfalfa plants and pea aphid abundance and phloem sap ingestion. This finding may indicate the importance of apigenin glycoside forms as nutritional compounds.
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