GroEL Gene And Double-Strand Rna Mediated Virus Resiatance To Tylcv In Tomato

Tomato(Lycopersicon esculentum Mill) is an important kind of vegetable crops in Jiangsu province.It plays a very important role in the year-round supply. However, tomao yellow curl disease (TYLCD) is so severe that the growth, flowering and fruit setting are damaged, thus, the quality and yield of tomato are influnenced. It has caused large losses in tomato production areas.Tomato yellow leaf virus (TYLCV) belongs to begomovirus Genera of Geminiviridae. It transmitted by the tobacco whitefly. The crop breeding of TYLCV resistance is limited by lacking the resistance resource, the presence of resistance resource in different areas and the diversity of virus genome caused by the hereditary isolation of different plant species, virus recombinant and variation. The occurrence regularity, resistance mechanism and resistant materials screening are founded by conventional breeding technology and biotechnology. In this research, tomato plants were transformed by transgenic approach to obtain resistance materials.In order to generate engineered resistance, we isolated the GroEL gene from Bemisa tabaci and constructed two different constructs:marker vector (SUC2-GroEL) and non-antibiotic gene marker vector (CaMV35S-PDRB10). At the same time, the inverted repeat sequence vector were constructed by using AC1(replication-relate) and AC2(movement) of TYLCV. The vectors were transformed into Agrobacterium tumefaciens, resistance of these transgenic tomato plants were evaluated. The results are shown as follows:1.The regenerated tomato plants of SUC2-GroEL and CaMV35S-PDRB10vector were obtained by Agrobacterium-mediated transformation. We obtained11plants of SUC2-GroEL vector and9plants of CaMV35S-PDRB10vector tomato plants resistant to kanamycin. The trangenic tomato plants were further analyzed the expression of GroEL gene at transcription level through RT-PCR, six plants of SUC2-GroEL vector and two plants of CaMV35S-PDRB10vector were both found to express in transcription level. The presence of trangene tomato plants by virus infection and viruliferous whiteflies inoculation were observed that three plants (3、4and5) of SUC2-Groel construct and one plant (5) of CaMV35S-PDRB10construct remained symptomless. These results suggest that it is feasible to use GroEL gene against TYLCV and the virus resistant of SUC2-GroEL vector is more effective than CaMV35S-PDRB10vector.2.The inverted repeat sequence vector pBI121-AC1-AC2was constructed by using AC1and AC2gene of TYLCV. The regenerated tomato plants were obtained by Agrobacterium-mediated transformation. We obtained13plants resistant to kanamycin. It confirmed that AC1and AC2repeat sequence had been inserted into tomato genome DNA. The presence of trangene tomato plants by virus infection and viruliferous whiteflies inoculation were observed that two plants remained symptomless.