Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: TILLING for allergen reduction and improvement of quality traits in peanut (Arachis hypogaea L.) | Knoll et al. BMC Plant Biology 2011 11 81 http 1471-2229 11 81 BMC Plant Biology RESEARCH ARTICLE Open Access TILLING for allergen reduction and improvement of quality traits in peanut Arachis hypogaea L. 1 1 2 2 3 Joseph E Knoll M Laura Ramos Yajuan Zeng C Corley Holbrook Marjorie Chow Sixue Chen Soheila Maleki4 Anjanabha Bhattacharya1 and Peggy Ozias-Akins1 Abstract Background Allergic reactions to peanuts Arachis hypogaea L. can cause severe symptoms and in some cases can be fatal but avoidance is difficult due to the prevalence of peanut-derived products in processed foods. One strategy of reducing the allergenicity of peanuts is to alter or eliminate the allergenic proteins through mutagenesis. Other seed quality traits could be improved by altering biosynthetic enzyme activities. Targeting Induced Local Lesions in Genomes TILLING a reverse-genetics approach was used to identify mutations affecting seed traits in peanut. Results Two similar copies of a major allergen gene Ara h 1 have been identified in tetraploid peanut one in each subgenome. The same situation has been shown for major allergen Ara h 2. Due to the challenge of discriminating between homeologous genes in allotetraploid peanut nested PCR was employed in which both gene copies were amplified using unlabeled primers. This was followed by a second PCR using gene-specific labeled primers heteroduplex formation CEL1 nuclease digestion and electrophoretic detection of labeled fragments. Using ethyl methanesulfonate EMS as a mutagen a mutation frequency of 1 SNP 967 kb 3 420 M2 individuals screened was observed. The most significant mutations identified were a disrupted start codon in Ara h and a premature stop codon in Ara h . Homozygous individuals were recovered in succeeding generations for each of these mutations and elimination of Ara h protein was confirmed. Several Ara h 1 protein isoforms were eliminated or reduced according to 2D gel analyses. TILLING
