RESUMEN
beta-1,3-glucanase(BG2) is one of the pathogensis-related-proteins(PR). Study of these proteins and their related genes is one of the hot points in plant genetic engineering of disease resistance for a long time. In this research, specific primers were designed with the enzyme cleavage site of Spe I in its forward one and Not I site in the backward according to the BG2 gene sequence. Using this pair of primers, BG2 gene, which was contained in the plasmid of pRTL2, was amplified and confirmed by sequencing the amplified fragment inserted into T-easy vector. The positive clone containing BG2 gene was digested with the enzymes of Spe I/Not I and then BG2 gene was inserted into the Xba I/Not I sites of super expression binary vector pATC940. The reconstructed expression vector named as pATCBG2 was introduced into the wheat of Longfumai10 and Longfumai3 (Triticum aestivum L. em. Thell) through the particle gun transformation method. The Kanamysin resistant (Km') transformants were obtained. PCR, Dot-blotting and PCR-Southern hybridization analysis showed that the BG2 gene was integrated into the genome of wheat. Result of pathogen inoculation assay on the transgenic plants showed that the transgenic plants had a higher resistant disease score of 1-2 grade than the control.
Asunto(s)
Vectores Genéticos/genética , Glucano 1,3-beta-Glucosidasa/genética , Triticum/genética , Ascomicetos/crecimiento & desarrollo , Southern Blotting , Clonación Molecular/métodos , ADN de Plantas/genética , ADN de Plantas/metabolismo , Desoxirribonucleasa EcoRI/metabolismo , Desoxirribonucleasa HindIII/metabolismo , Inmunidad Innata/genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Plantas Modificadas Genéticamente , Transformación Genética , Triticum/microbiologíaRESUMEN
Southern corn rust (SCR) is a destructive disease in maize. The inbred line Qi319 is highly resistant to southern corn rust. SSR technique was employed to preliminary mapping of the resistance gene. Bulked segregant analysis revealed that two primers, phi 118 and phi 041, amplified polymorphic bands. SSR analysis on populations indicated the two primers were linked to the rust resistance gene, which was mapped on the short arm of chromosome 10. In addition, comparative analysis of the amplification bands among different populations revealed that the amplification products with the same primer in different populations were dissimilar. This result indicates that the genetic background may affect results of gene mapping and tagging. So, it is important to select suitable population to performing molecular marker analysis and gene mapping.
Asunto(s)
Basidiomycota/crecimiento & desarrollo , Mapeo Cromosómico/métodos , Zea mays/genética , Cromosomas de las Plantas/genética , ADN de Plantas/genética , Inmunidad Innata/genética , Repeticiones de Microsatélite/genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Polimorfismo de Longitud del Fragmento de Restricción , Zea mays/microbiologíaRESUMEN
Study of beta-1,3-glucanase is one of the hot points in plant genetic engineering of disease resistance,big progress has been made in the past few years. This paper briefly reviewed the main biological characters and mechanism involved in the disease resistance,the classification,structure,function and the transformation of beta-1,3-glucanase genes.