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This thesis consists of two parts. (A) bisporus browning polyphenol oxidase (PPO) (1) bisporus during storage prone to browning affect its food value and economic value. In this study, through research bisporus browning during storage, total phenolic content, polyphenol oxidase (PPO) and benzene alanine ammonia lyase (PAL) enzyme activity changes, explore the Browning Mechanism to find suppression its browning ways. The results showed that the PPO bisporus browning closely linked, but the browning mechanism needs further study. (2) is separated from the bisporus and cloned two PPO homologous genes in genomic sequence of PPO to 3 and PPO4, length of 2080bp and 2189bp, respectively, the sequence information was analyzed and found that PPO ~ 3 and PPO4 the connection zone 7 exons and 6 introns, exons and introns are highly conserved eukaryotic gene GT-AG splicing rule, and contains a typical CuA, CuB domain. (3) the use of technology were cloned two PPO gene promoter sequence length greater than 1.5 kb chromosome walking analysis found that contains the core promoter of conservative elements, such as TATA-Box and CAAT-Box, and I-box , G-Box, AE-box, WUN-motif, ARE-motif, MYB some important cis-acting elements. (4) The semi-quantitative results proved once again that the relationship between the PPO gene bisporus browning. (B) the establishment of the mushrooms EST-SSR markers EST-SSR markers based on EST or cDNA sequences in public databases developed a molecular marker. EST-SSR markers development mushrooms available on the genetic breeding of edible fungi of great significance. This study bioinformatics data mining techniques, the identification and analysis of public sequence databases mushrooms EST SSR loci, established mushrooms EST-SSR markers, and markup application shiitake spawn genetic relationship obtained the following results: (1) downloaded from NCBI 12,184 the mushrooms EST sequences, full-length 3681177bp 4684 Unigene treated. Excavations in which a total of 142 SSR distribution in 120 Unigene occurrence frequency of 2.99.% (1/25.429kb). , Single, two, trinucleotide repeat major repeat type, A / T, AG / CT, ACG / CTG is a single, two, three nucleotides repeated motif, accounted for all of EST-SSR 78.57%, 79.49%, and 25.42%, respectively. (2) using primer5.0 40 primer is designed, and in the 35 mushroom species detected. The results 36 primer amplification, accounting for 90% of all primers, 22 pairs of primers for polymorphic primers polymorphic rate was 55%. (3) application NTSYS software clustering analysis showed that 35 were genetic similarity coefficient range of 0.125-0.781 PIC = 0.056 - 0.826, 35 mushrooms species have certain genetic diversity. Can be further applied to the practical application of the mushrooms genetic breeding.
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