Identifikacija alela gena za puroindolin i njihov utjecaj na teksturu zrna pšenice (Triticum aestivum L.)

Grain hardness is one of the most important quality characteristics of wheat (Triticum aestivum L.). It is a significant property of wheat grains and relates to milling quality and end product quality. Grain hardness is caused by the presence of puroindoline genes (Pina and Pinb). A collection of 25 genotypes of wheat with unusual grain colour (blue aleurone, purple and white pericarp, yellow endosperm) was studied by polymerase chain reaction (PCR) for the diversity within Pina and Pinb (alleles: Pina-D1a, Pina-D1b, Pinb-D1a, Pinb- -D1b, Pinb-D1c and Pinb-D1d). The endosperm structure was determined by a non-destructive method using light transfl ectance meter and grain hardness by a texture analyser. Genotype Novosibirskaya 67 and isogenic ANK lines revealed hitherto unknown alleles at the locus for the annealing of primers of Pinb-D1. Allele Pinb-D1c was found to be absent from each genotype. The mealy endosperm ranged from 0 to 100 % and grain hardness from 15.10 to 26.87 N per sample.Tvrdoća zrna jedno je od najvažnijih svojstava pšenice (Triticum aestivum L.) koje utječe na mlinarsku kakvoću i svojstva krajnjeg proizvoda, a određuju ju geni za puroindolin (Pina i Pinb). Metodom lančane reakcije polimeraze određena je prisutnost različitih alela gena Pina i Pinb (Pina-D1a, Pina-D1b, Pinb-D1a, Pinb-D1b, Pinb-D1c i Pinb-Did) u 25 genotipova pšenice neobične boje zrna (plavi aleuron, ljubičasti i bijeli perikarp, žuti endosperm), a nakon toga su ispitani struktura i tvrdoća zrna. U genotipu Novosibirskaya 67 i izogenoj liniji ANK otkriveni su dosad nepoznati aleli na mjestu sparivanja početnica gena Pinb-D1. Utvrđeno je da genotipovi nisu sadržavali alel Pinb-D1c. Udjel je brašnastog endosperma u uzorku zrna pšenice bio od 0 do 100 %, a tvrdoća zrna od 15,10 do 26,87 N

Testing of DNA isolation for the identification of hemp

Hemp is diploid organism (2n = 2x = 20, genome size 534 Mb) with nine pairs of autosomes plus XX (♀) or XY (♂) chromosomes. Cannabis sativa L. is an important economic plant for the production of food, fibre, oils, and intoxicants. Genotypes (varieties or chemovar) of hemp with low Δ9-tetrahydrocannabinol content are used for industrial applications. Varieties with high Δ9-tetrahydrocannabinol or high cannabidiol content are used for medicinal applications. Biochemical and molecular methods can be used for identification and classification. An important step for molecular biology methods is to obtain the matrix of the native and sufficiently pure DNA. We tested two different experimental variant of samples (20 mg and 100 mg) of seeds, oilcake and dried flowers for analysis of the Italian variety Carmagnola for analysis (harvested in 2014, Hempoint Ltd., Czech Republic). The DNeasy® Plant Mini Kit (Qiagen, GE) was used to isolate the DNA. The DNA concentration and purity was assessed by agarose electrophoresis and via a spectrophotometer. Samples of lower weight yielded lower values of DNA concentration (average 16.30 - 38.90 ng.µL-1), but with better purity than samples of higher weight (ratio A260nm/A280nm for low-weight samples was near 1.80). To test the applicability of DNA analysis, we used two SSR markers (CAN1347 and CAN2913). PCR products were separated on 1% agarose and on 8% polyacrylamide electrophoresis. DNA samples obtained from samples of higher weight exhibited less PCR amplification than samples of lower weight. We found no effect of sample weight on the formation of non-specific amplification products during the PCR reaction. Based on our results we can be recommended for practical isolation procedure using DNeasy® Plant Mini Kit with lower of sample weight (20 mg). In future work the procedure for DNA isolating from wheat-cannabis products, e. g. breads, rolls or pasta, will be optimized

The effect of feeding wheat with purple pericarp on the growth of carp

This study assessed and compared the influence of feeding wheat with purple pericarp (variety Konini) and standard coloured wheat (red variety Bohemia) on the growth characteristics of fingerling carp (Cyprinus carpio L.) of the "Amurský lysec" line. The total content of anthocyanins converted to cyanidin 3-glucoside in the control Bohemia wheat was 24.95 mg.kg-1 and in the Konini purple wheat 41.70 mg.kg-1. Two experimental variants for feed were evaluated: dipped wheat grain and crushed wheat grain. The feed dose for wheat was 1.5% of the fish stock weight and for natural food (frozen Chironomid larvae) was 0.2% of fish stock weight to all variants. Growth parameters (body length, body weight, Fulton's condition factor and feed conversion ratio) of the fish were evaluated after one month of administration. The feed consumption and physico-chemical parameters (temperature, oxygen saturation, pH, N-NH4 +, N-NO2-, N-NO3- and Cl-) of the environment were observed. During the feeding test, no major differences in food consumption among variations feeding on either wheat and on Chironomid larvae were noted. Satisfying results for mas and length gain were achieved in V2 wheat with purple pericarp (Konini variety - dipped grain), where the average total body length was 156.56 mm and the average unit mass was 60.81 g. In this variant, higher values of the parameters were achieved compared to the control group (100.6%, resp. 104.2%). A positive impact of wheat with purple pericarp on the evaluated parameter of fish condition factor was demonstrated. This trend was confirmed in all variants. No effect was demonstrated for mechanical disruption of kernels on the level of utilization of nutrients. In further experiments on growth characteristics we would like to determine antioxidant parameters in the blood and liver of fry

Identification of Alleles of Puroindoline Genes and Their Effect on Wheat (Triticum aestivum L.) Grain Texture

Grain hardness is one of the most important quality characteristics of wheat (Triticum aestivum L.). It is a significant property of wheat grains and relates to milling quality and end product quality. Grain hardness is caused by the presence of puroindoline genes (Pina and Pinb). A collection of 25 genotypes of wheat with unusual grain colour (blue aleurone, purple and white pericarp, yellow endosperm) was studied by polymerase chain reaction (PCR) for the diversity within Pina and Pinb (alleles: Pina-D1a, Pina-D1b, Pinb-D1a, Pinb- -D1b, Pinb-D1c and Pinb-D1d). The endosperm structure was determined by a non-destructive method using light transfl ectance meter and grain hardness by a texture analyser. Genotype Novosibirskaya 67 and isogenic ANK lines revealed hitherto unknown alleles at the locus for the annealing of primers of Pinb-D1. Allele Pinb-D1c was found to be absent from each genotype. The mealy endosperm ranged from 0 to 100 % and grain hardness from 15.10 to 26.87 N per sample