Construction of a BAC library for buckwheat genome research - an application to positional cloning of agriculturally valuable traits.
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We have constructed a BAC library for common buckwheat Fagopyrum esculentum Moench. The library includes 142,005 clones with an average insert size of approximately 76 kb, equivalent to approximately a 7 to approximately 8-fold coverage of the genome. Polymerase chain reaction based screening of the library with AGAMOUS and FLORICAULA/LEAFY primers, has identified 7 and 9 BACs, respectively, which are consistent with the genome coverage. This library represents the first large insert genomic library for F. esculentum and it can be served as a genetic resource facilitating agricultural, pharmacological, physiological, and evolutionary studies of the species. To demonstrate the utilization of the library for characterizing agriculturally valuable traits, we developed a sequence tagged site marker tightly linked to the dwarf E locus as well as to the self-incompatibility complex locus and screened the library to initiate positional cloning of the causative genes. (+info)
Use of buckwheat seed protease inhibitor gene for improvement of tobacco and potato plant resistance to biotic stress.
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The possibility to use agrobacterial transformation of leaf discs to produce resistance to bacterial infections in tobacco and potato plants by introduction of a single gene encoding the serine proteinase inhibitor BWI-1a (ISP) from buckwheat seeds is shown. All studied PCR-positive transgenic plants exhibited antibacterial activity in biotests. It was shown that the presence of just a single gene of serine proteinase inhibitor provides sufficient protection at least against two bacterial phytopathogens, Pseudomonas syringae pv. tomato and Clavibacter michiganensis sbsp. michiganensis. The biotest including tobacco plant infection by the white wings butterfly in the green house has also demonstrated the existence of protective effect in transgenic tobacco plants. Significant genotypic variations in the protection efficiency were found between members of different genera of the same family (potato and tobacco) as well as between different lines of the same species. Northern blot analysis of four transgenic potato lines and three tobacco lines transformed by a vector plasmid containing the ISP gene of serine proteinases BWI-1a from buckwheat seeds has shown the presence of the expected size mRNA transcript. (+info)
Selenium and its species distribution in above-ground plant parts of selenium enriched buckwheat (Fagopyrum esculentum Moench).
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Common buckwheat (Fagopyrum esculentum Moench) was foliarly sprayed with a water solution containing 10 mg Se(VI) L(-1) at the beginning of flowering. The total Se content in plant parts in the untreated group was low, whereas in the Se-sprayed group it was approximately 50- to 500-fold higher, depending on the plant part (708-4231 ng Se g(-1) DM(-1) (DM: dry matter)). We observed a similar distribution of Se in plant parts in both control and treated groups, with the highest difference in Se content being in ripe seeds. Water-soluble Se compounds were extracted by enzymatic hydrolysis with protease XIV, resulting in above 63% of soluble Se from seeds, approximately 14% from stems, leaves and inflorescences and less than 1% from husks. Se-species were determined in enzymatic extracts using HPLC-UV-HG-AFS (HPLC-hydride generation-atomic fluorescence spectrometry with UV treatment). The main Se species found in seeds was SeMet ( approximately 60% according to total Se content), while in stems, leaves and inflorescences the only form of soluble Se present was Se(VI) (up to 10% of total Se). In husks no Se-species were detected. We observed an instability of Se(IV) in seed extracts as a possible consequence of binding to the matrix components. Therefore, special care concerning sample extraction and the storage time of the extracts should be taken. (+info)
Purification, crystallization and preliminary X-ray analysis of a deletion mutant of a major buckwheat allergen.
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Electromyographic measurement of eating behaviors for buckwheat noodles.
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The objective of this study was to analyze human eating behaviors in chewing and slurping buckwheat noodles. We used electromyography to measure the activity of the jaw-closing, jaw-opening, and lip-closing muscles while healthy adults ate one mouthful of buckwheat noodles. Slurping the noodles required a longer mastication period but smaller muscle activity per movement than chewing the same samples. Total muscle activity was greater in slurping. Slurping also showed a longer average cycle time but greater variances in the cycle time than rhythmical chewing. The mechanical properties of buckwheat noodles significantly differed between the noodle types (half-raw and dry), but the human mastication variables for the two types of noodles were not significantly changed within a subject. Both types of noodles kept for 10 min at 23 degrees C after being cooked could be consumed with less mastication effort than those immediately served, and this observation corresponded to softening of the noodles during the standing time. (+info)
Light stimulation triggered expression of genes coding for vacuolar proton-pump enzymes V-ATPase and V-PPase in buckwheat.
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Although coloration in plants is ascribable to both the accumulation of anthocyanin pigments in vacuoles and to the acidification of vacuolar pH, the environmental factors causing the decrease in vacuolar pH are unknown. We found that blue-light irradiation of buckwheat seedlings using light-emitting diodes caused reddening on the surface of the hypocotyls. It has also been reported that light stimulation induces an accumulation of anthocyanin pigments. However, here we confirmed for the first time on the basis of real-time PCR analysis that light stimulation simultaneously triggers expression of the genes coding for subunit A of vacuolar H+-ATPase (V-ATPase) and vacuolar H(+)-pyrophosphatase (V-PPase). (+info)
Genotypic differences in Al resistance and the role of cell-wall pectin in Al exclusion from the root apex in Fagopyrum tataricum.
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