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The aim of this research was to characterize somaclonal variation in populations derived from embryos cultured on two types of induction medium (supplemented with either 2,4-D or dicamba), as well as to select and characterize several somaclonal lines. The sexual progenies of 40 R0 regenerants - A somaclones (derived on the medium with 2,4-D) and B somaclones (derived on the medium with dicamba) - were analysed according to the following traits: plant height, total number of tillers, number of productive tillers, spike length, number of spikelets per spike, spike compactness, number of normally developed grains per spike, weight of grains per spike, and the weight of 1000 grains. The results for twenty-two R1 plants surpassed the variability range for the control. The transmission of positive changes to the next generation was proved in the case of 8 originally chosen R1 plants: 7 plants selected from the A somaclones and one plant from the B somaclones. Five out of the eight created somaclonal lines proved to be stable somaclonal variants. The absolute rate of the efficiency of positive somaclonal changes was calculated as 0.64%.
Several methods of transformation are currently available for delivering exogenous DNA to plant cells. Agrobacterium-mediated transformation, microprojectile bombardment and direct protoplast transformation are routinely used today. However, each of them has certain disadvantages, which led to research into the development of novel alternative systems such as infiltration, electroporation of cells and tissues, electrophoresis of embryos, microinjection, pollen-tube pathway, silicon carbide- and liposome-mediated transformation. The low efficiency of transformation is considered to be the main reason for the limited popularity of the alternative transformation methods, other than infiltration and silicon carbide-mediated transformation, which seem to be the most promising ones for practice.
The genetic basis of the regeneration process in cultured immature embryos of rye (Secale cereale L.) was analyzed. The experiments were designed to reveal differences between the in vitro culture responses of two inbred lines: L318 (a high regeneration ability) and L9 (a low potential for regeneration). The rye ortologues of plant genes previously recognized as crucial for somatic embryogenesis and morphogenesis in vitro were identified. Using oligonucleotide primers designed to conserved regions of the genes Somatic Embryogenesis Receptor-like Kinase (SERK), Leafy Cotyledon 1 (LEC1), Viviparous 1 (VP1) and NiR (encoding ferredoxin-nitrite reductase), it was possible to amplify specific homologous sequences from rye RNA by RT-PCR. The transcript levels of these genes were then measured during the in vitro culture of zygotic embryos, and the sites of expression localized. The expression profiles of these genes indicate that their function is likely to be correlated with the in vitro response of rye. In line L9, increased expression of the rye SERK ortologue was observed at most stages during the culture of immature embryos. The suppression of ScSERK expression appears to start after the induction of somatic embryogenesis and lasts up to plant regeneration. The rye ortologues of the LEC1 and VP1 genes may function in a complimentary manner and have a negative effect on the production of the embryogenic callus. The expression of the rye NiR ortologue during in vitro culture reveals its importance in the process of plant regeneration.
Microsatellites (SSR - simple sequence repeats, STR - short tandem repeats, SSLP - simple sequence length polymorphism, VNTR - variable number of tandem repeats) are the class of repetitive DNA sequences present in all living organisms. Particular characterstics of microsatellites, such as their presence in the genomes of all living organisms, high level of allelic variation, co-dominant mode of inheritance and potential for automated analysis make them an excellent tool for a number of approaches like genotyping, mapping and positional clonig of genes. The three most popular types of markers containing microsatellite sequences that are presently used are: (1) SSR (simple sequence repeats), generated by amplifying in a PCR reaction with the use of primers complementary to flanking regions; (2) ISSR (inter-simple sequence repeats), based on the amplification of regions between inversely oriented closely spaced microsatellites; and (3) SAMPL (selective amplification of microsatellite polymorphic loci), which utilises AFLP (amplified fragment-length polymorphism) methodology, with one exception - for the second amplification, one of the starters is complementary to the microsatellite sequence. The usefulness of the three above-mentioned markers for numerous purposes has been well documented for plants.
This study was conducted in order to identify quantitative trait loci (QTLs) for the in vitro culture response of winter rye (Secale cereale L.) immature embryos and immature inflorescences. A genetic linkage map comprising 67 SSRs, 9 ISSRs, 13 SAMPLs, 7 RAPDs, 2 SCARs and one EST marker was created based on the analyses of 102 recombinant inbred lines from the cross between lines L318 (which has a good response in tissue cultures) and L9 (which is unable to regenerate plants from somatic tissues and anthers). The map spans 979.2 cM, and the average distance between markers is 9.9 cM. Two characteristics were evaluated: callus induction (CI) and somatic embryogenesis ability (SE). They were expressed as the percentage of immature embryos/inflorescences producing callus (designated ECI/ICI) and the percentage of explants producing somatic embryos (ESE/ISE). All the analysed traits showed continuous variation in the mapping population but a non-normal frequency distribution. We identified nine putative QTLs controlling the tissue culture response of rye, explaining up to 41.6% of the total phenotypic variation: two QTLs for ECI — eci-1, eci-2; 4 for ESE — ece-1, ese-2, ese-3, ese-4; 2 for ICI — ici-1, ici2; and 1 for ISE — ise-1. They were detected on chromosomes 1R, 4R, 5R, 6R and 7R.
Benzoxazinoids (BXs) are important compounds in plant defense. Their allelopathic, nematode suppressive and antimicrobial properties are well known. BXs are found in monocot plants and in a few species of dicots. Over 50 years of study have led to the characterization of the chromosomal locations and coding sequences of almost all the genes involved in BX biosynthesis in a number of cereal species: ZmBx1–ZmBx10a7c in maize, TaBx1–TaBx5, TaGT and Taglu in wheat, ScBx17ScBx5, ScBx6-like, ScGT and Scglu in rye. So far, the ortholog of the maize Bx7 gene has not been identified in the other investigated species. This review aims to summarize the available data on the genetic basis of BXs biosynthesis in cereals.
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