Influence Of Infection By Paulownia Witches'broom-phytoplasma On Enzyme Activities Of Paulownia In Vitro Cultured Plantlets And The Clone Of Related Gene Fragment

This paper comprises three parts. The first part is about the influences on activities of paulowniaplantlet' s POD and IAAO by exterior reagents. The second part is about purification of enzyme related to the infection of paulowina witch' s broom-phytoplasma. The third part is about the clone of POD gene fragment related to the infection of paulowina witch' s broom-phytoplasma.The healthy and diseased paulownia plantlets were treated by exterior plant growth regulators (NAA and 6-BA) and a kind of phenol (SA). The results as following: one week after the treatment of NAA, fitable concentration could decrease the activities increase of POD caused by hurt . Two and four weeks later, the activities of POD were increased as the NAA concentration thicked in generally .There were large differences between two clones in the change of IAAO activities .The plantlets grew better after treatment of NAA. When treated with 6-BA , the activities of POD and IAAO decreased in every week .The healthy plantlets appeared diseased symptoms and the diseased one appeared heavier symptoms .When treated with SA the activities of POD increased but the activities of IAAO decreased in both healthy and diseased plantlets .At the range of treatment concentration , SA could restrain the growth of healthy planglets but accelerated diseased ones .The activities of POD were increased after phenols were removed with PVP-P but IAAO decreased a little .Using the plantlets of paulownia, the specific protein had been iaolated .The enzyme purification steps included homogenization , (NH4)2SO4 precipitation and consective chromatography on DEAE-Sephadex A50 and CM-52. This protein is specific increased POD in diseased plantlet.The result of HPLC indicated that the content percent is more than 99. This protein was characterized having a specific activity of 371U/mg .A fragment of induced expression of POD gene was cloned from diseased paulownia plantlets. This fragment of POD gene was derived from diseased plantlets but healthy one not.The length was 871 bp .This sequence is a 3' -terminal fragment without inexon . It was predicted from these sequence that the protein was synthesized of 219 amino acid residues.The theoretical Mr was 24074 Da. This putative protein probably is the result of infection of phytoplasma .