Purification method for grafting and inoculating of tomato chlorotic virus by taking tobacco as rootstock and application

Fecha de publicación: 27/03/2020
Fuente: Wipo tomato
The invention belongs to the technical field of plant virus purification, and discloses a purification method for grafting and inoculating of tomato chlorotic virus by taking tobacco as a rootstock and application of the purification method, which comprises the following steps of: grafting a scion onto the rootstock by taking diseased Nicotiana benthamiana as the rootstock and taking a tomato plant as the scion by utilizing a cleft grafting method; and after the scion tomato grows to a leaf stage, cutting off from the tomato root, reserving the overground part as the scion, obliquely cutting apart of phloem at each of the left and right sides of the lower part of the scion tomato, inserting into a Nicotiana benthamiana plant incision, and fixing by using a grafting clamp. The ToCV infective clone disclosed by the invention can successfully infect the Nicotiana benthamiana and is obvious in morbidity symptom, the ToCV infective clone is directly inoculated to a tomato plant, the infection rate is extremely low, no symptom is shown, and no virus fragment is detected in vivo after bemisia tabaci eats the tomato plant for 48 hours; the incidence rate of the tomato plants grafted by the cleft grafting method is as high as 79.08%, and the method is more suitable for grafting the tomato plants with the nicotiana benthamiana as the rootstocks.