Function Researches Of Rtps From Bouganinvillea Glabra In Transgenic Tomatoes To Improve The Resistances To Diseases And Insects

Ribosome inactivating proteins(RIPs)are a kind of defensively toxic protein widely found in higher plants.The glycosidic bond is specifically hydrolyzed by RNA N-glycosidase activity at the A4234 position between ribose and adenine base of ribosomes and then the ribosome structure is destroyed to achieve the purpose of inhibiting protein synthesis.RIPs have a wide range of polymorphisms and a variety of biological functions.At present,RIPs have been paid attention to the research and application of anti-cancer,anti-virus and other medical fields,while their resistance to plant diseases and insect pests are widely reported.In this study,Bougainvillea was used as material.Primers were designed according to the cDNA sequence of RIPs gene of Bougainvillea spectabilis Willd,which was reported in the literature.Ten fragments of RIPs homologous gene were amplified by RT-PCR using B.glabra Choisy cDNA as template.The fragments were named separately and the number of Bou-J was identical to the original fragment that had been amplified in the early stage of the laboratory,and Bou-K has same sequence as the B.spectabilis Willd RIPs gene which reported in the literature.The fragments of Bou-J and Bou-K were selected by constructing the expression vector,and the target gene fragments Bou-J and Bou-K were introduced into tomato plants by Agrobacterium tumefaciens.We obtainded 18 strains of Bou-J and 6 strains of Bou-K positive transgenic tomato plants after the positive identification.We conducted a preliminary study on the resistance to diseases and pest insects of transgenic tomato plants.After the study,we get the following results:Bou-J and Bou-K transgenic tomato plants showed significant resistance to aphids,and Bou-J group was stronger than Bou-K group.Bou-J and Bou-K transgenic plants showed resistance to botrytis cinerea and the resistance of the two transgenic plants was basically same.Bou-K transgenic plants showed the resistance to tobacco mosaic virus,while the Bou-J group did not show any resistance to tobacco mosaic virus.This study shows that:the resistance of transgenic tomato confirmed Bougainvillea RIPs have the resistance to insect and properties,which can be applied to plant transgenic breeding for disease and insect pest.Bougainvillea RIPs have more resistance at the same time,as Bougainvillea RIPs in the plant genetic resources and breeding new plant varieties resistant to disease and insect pests have provided the theoretical basis and guidance.In this study,a number of RIPs homologous gene fragments were obtained,which was consistent with the previous study on the polymorphism of Bougainvillea RIPs gene of our laboratory.Bou-J and Bou-K transgenic tomatoes were different,which indicated that the RIPs function was also polymorphic.Therefore,when we use the Bougainvillea RIPs for plant transgenic breeding,we should screen the target genes to obtain transgenic plants with strong resistance and antibacterial and insect-resistant wide spectrum.