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The effect of explant type on somatic embryogenesis induction in Pisum sativum (cv. Oskar and an unregistered line HM-6) was studied. Shoot apices, leaf primordia, and epicotyl fragments of axenically grown, etiolated seedlings, as well as embryonic axes and cotyledon fragments isolated from zygotic embryos at different stages of development, were used as explants. Somatic embryogenesis was induced essentially as described by Griga in 1998 – MS salts and sucrose, B5 Gamborg vitamins, picloram (2.5 μM). After induction period (14 days) all cultures were transferred to the differentiation medium (basal medium as above, auxin omitted). Both in Oskar and HM-6, only shoot apices developed somatic embryos and (with significantly lower frequency) adventitious shoots.
The aim of this study was to compare the physiological responses of six plant species (popular crops or plants recommended as indicators of soil pollution) to a wide range of glyphosate concentrations (0, 1, 3, 7, 10, 40, 80, 120, 180, 240, 400, 750, 1,000, 1,500, 1,700 and 2,000 μM). Percent germination, root length, seedling dry mass and myo-inositol content, as well as seedling leachate electroconductivity were determined in Lepidium sativum, Sinapis alba, Sorghum saccharatum, Brassica napus, Lupinus luteus and Avena sativa. Percent seed germination, seedling dry mass and electroconductivity of seedling leachates were not clearly affected by the herbicide and could not be used as indicators of its phytotoxicity. An metabolite induced by abiotic stresses in many plants, myo-Inositol, was very strongly stimulated by glyphosate at doses above 10 or 40 μM, depending on plant species. The sensitivity of analyzed plants to glyphosate, as manifested by root length, differed clearly. In Avena sativa the relationship between root length and glyphosate concentration was fairly linear over a wide range of herbicide doses (up to 240-400 μM). The most distinct drop in root growth at low herbicide doses was visible in Sorghum saccharatum. The results show that a mild stress affecting root length may not clearly modify seedling myo-inositol levels, that respond distinctly to stronger stresses. Not all indicator plants are equally suitable for analysis of biological activity of glyphosate residues. Sorghum saccharatum seems particularly sensitive.
Seeds of Lathyrus sativus cv. Derek and Krab were used as biological material for induced mutagenesis. Three mutant lines were obtained from seeds of grass pea cv. Derek and 15 lines from mutagenised seeds of cv. Krab. Twelve ethanol-soluble carbohydrates were identified in the seeds. We have selected grass pea mutant lines with high oligosaccharides content (lines D4, K56, K25, and K7) and lines with low raffinose family oligosaccharides (RFO) content (lines K12, K29 and K13). Mutations changing the levels of RFO have not affected the contents of galactosyl cyclitols.
W pracy oceniono żywotność oraz zawartość węglowodanów rozpuszczalnych w nasionach bobiku, grochu, łubinu i soczewicy przechowywanych przez 22 lata w temperaturze -14°C, 0°C oraz w temperaturze pokojowej (około +20°C). Przechowywanie nasion w temperaturze pokojowej powodowało, że nasiona nie kiełkowały, natomiast przechowywane w temperaturze -14°C i 0°C kiełkowały od 86 do 98% w zależności od gatunku. Nasiona przechowywane w temperaturze +20°C miały niższą, w porównaniu do nasion przechowywanych w temperaturze -14°C, 0°C, zawartość oligosacharydów rodziny rafinozy. Podczas przechowywania węglowodany te ulegały degradacji, a proces ten obejmował przede wszystkim werbaskozę i stachiozę.
The gene fusion system was used to study UV light-control of PS PAL1 and PS PAL2 genes encoding phenylalanine ammonia-lyase of pea. The induction of pea PAL promoters was analysed in transgenic tobacco plants. Binary plasmids (derivatives of pBI101.2 vector) containing 5’ regulatory fragments of PS pAl1 and PS PAL2 linked to reporter genes (GUS, LUC) were constructed. The analyses were performed with the use of single constructs (containing one variant of PS PAL promoter and one reporter gene) and dual constructs (containing both PS PAL1 and PS PAL2 promoters connected with different reporter genes). The use of dual constructs enabled the evaluation of both PS PAL promoters activity in the same plant. The analyses of in vitro grown plants have shown that both PAL promoters are strongly induced in leaves subjected to UV radiation. In some cases, the UV-stimulated expression exceeded the exposed areas. This phenomenon was observed more often in the leaves of plants containing the PS PAL1:: GUS than PS PAL2::GUS construct. Removal ofboxes 2, 4, 5 from PS PAL1 promoter and deletion of its 5’end region (-339 to -1394) decreases the level of gene expression but does not eliminate its responsiveness to UV.
The aim of this study was to compare the accumulation of soluble carbohydrates in embryos of two lupin species: cultivated Lupinus luteus (cv. Juno) and wild L. pilosus, developing on plants grown under normal soil humidity and soil drought. All analysed seeds accumulated soluble carbohydrates, including: monosaccharides, sucrose, cyclitols, galactosyl cyclitols and raffinose family oligosaccharides. Soil drought caused a nearly two-fold increase of soluble carbohydrate contents in both species. L. pilosus embryos however, responded to water deficiency by increasing the accumulation of cyclitols and galactosyl cyclitols, whereas L. luteus embryos enhanced accumulation of cyclitols and raffinose family oligosaccharides.
Germinability and the content of soluble carbohydrates were analysed in cereal seed (winter rye, cv. Warko; spring wheat, cv. Santa; hexaploid winter triticale, cv. Fidelio and cv. Woltario). Seed moisture content (mc) was equilibrated over silica gel to 0.08 g H2O/g dry mass and stored in a desiccator at 20oC for up to 205 weeks or were equilibrated to mc 0.06, 0.08 or 0.10 g H2O/g dm and subjected to artificial aging at 35oC in air-tight laminated aluminium foil packages for 205 weeks. It was shown that the rate of seed aging depended on the species and seed moisture content. The fastest decrease of germinability upon storage was observed in seed with the highest moisture level. Complete germinability loss for winter rye, winter triticale cv. Fidelio, winter triticale cv. Woltario and spring wheat seed with mc 0.10 g H2O/g dm3 occurred after 81, 81, 101 and 133 weeks, respectively. Fructose, glucose, galactose, myo-inositol, sucrose, galactinol, raffinose, stachyose and verbascose were the main soluble carbohydrates found in the seed. The obtained data on the contents of specific sugars and the composition of soluble sugars fraction in seed of rye, wheat and triticale did not corroborate any profound effect of reducing sugars, sucrose and oligosaccharides on seed longevity.
Physiological parameters (vigour and viability, total protein, albumin and globulin contents) were studied in yellow lupin seeds stored for 20 years at -14ºC, 0ºC and at room temperature (approx. 20ºC). The storage of yellow lupin cv. Iryd seeds, at room temperature resulted in viability decrease to 2% and increased seed leachate. No evident differences could be observed in vigour and viability of seeds stored at lowered (0ºC and -14 ºC) temperatures. Protein content was the lowest in seeds stored at +20 ºC and the highest in seeds stored at -14ºC.
Seed vigour, viability, the contents of soluble carbohydrates, total protein, albumins, and globulins, as well as seed coat structure, were analysed in yellow lupin (Lupinus luteus L.) cv. Iryd seeds stored for 20 years at -14oC, 0oC or at room temperature (approx. +20oC). Seed storage at room temperature reduced viability (to 2%) and increased seed leachate electroconductivity. Determinations of total proteins showed that protein content was significantly reduced in seeds stored at +20oC compared to the other storage regimens. Raffinose family oligosaccharides were the main soluble carbohydrates in seeds stored at 0oC and -14oC, whereas sucrose dominated in seeds stored at room temperature. Scanning electron microscopy (SEM) of seed surface and seed coat sections revealed appearance of an amorphic layer on the surface of seeds stored at room temperature (not observed in other seeds) and distinct shrinking of macrosclereid layer in seeds stored at -14oC. Macrosclereids layer in all seeds was 100 um thick and accounted for 60% of seed coat thickness. The obtained results suggest that for long term storage of lupin seeds at 0oC is the most advisable temperature if both costs of storage and seed storability are considered.
Seed sculpture, seed coat thickness and soluble carbohydrate profiles were compared in three lupin species, each represented by three different genetic lines, from sections Pilosus (L. pilosus) and Atlanticus (L. atlanticus, L. cosentinii). Seed coat thickness varied most in the three lines of L. pilosus (330-1000 µm), while in the other species it varied little from 400 µm thickness. The studied seeds accumulated approximately 100 mg/g d.m. soluble carbohydrates. The obtained results together with published data suggest that the soluble carbohydrate composition of seeds is clearly section-specific in Lupinus sections Pilosus and Atlanticus. Sect. Pilosus was characterized by a high level of galactosyl cyclitols, while seeds of sect. Atlanticus contained much higher amounts of the raffinose-family oligosaccharides. In all species, however, the dominating soluble carbohydrate was stachyose. The results on the biochemical parameters analyzed in this paper confirm that L. atlanticus and L. cosentinii are more closely related to each other than to L. pilosus. The seed coat roughness/smoothness criterion seems inadequate for building a classification of lupin species, as one of the analyzed lines of L. pilosus, formally ascribed to the rough-seeded lupins, produces smooth seeds.
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