Method for observing transferring of fusion protein to rice leafbud at long distance

Abstract

The invention discloses a method for observing the transferring of a fusion protein to a rice leafbud at long distance. The method comprises the following steps of: treating a root tip of rice by using a GFP (Green Fluorescent Protein) fusion protein; then, freezing a leafbud of a rice plant with a treated root tip, and slicing; next, dyeing by using a PI (Propidium Iodide) fluorescent dye; and directly observing the transferring condition of the GFP fusion protein from the root tip to the rice leafbud by using a confocal microscopy. By using the method, the position, 0.5cm away from the root tip, of the rice is treated by using the GFP fusion protein, the leafbud is frozen and sliced and then dyed, and the transferring condition of the GFP fusion protein from the root tip to the rice leafbud is observed by using the confocal microscopy, so that the long-distance transferring condition of the GFP fusion protein in the rice plant can be directly and qualitatively known about. The position, 0.5cm away from the root tip of the rice, is directly treated by using the GFP fusion protein, the transferring of the GFP fusion protein to the leafbud is observed by using the confocal microscopy, and thus, a very powerful evidence can be further provided for the fact that an effect protein can induce the defensive reaction of a rice system. The method is simple, convenient, accurate and rapid.

Description

technical field [0001] The present invention relates to a method of using GFP fusion protein to treat rice root tips to observe the long-distance transport of the fusion protein to rice leaf buds, and in particular to a method for treating rice root tips at 0.5 cm by using the prokaryotic expression product of the rice blast fungus protein gene fused with GFP, The invention discloses a laser confocal microscopy method for observing the translocation of GFP fusion protein from root tip 0.5cm to rice leaf buds, belonging to the fields of plant protection and cell biology. Background technique [0002] Pathogenic bacteria transport effectors through the type III secretion system, and its transport mechanism has been studied relatively clearly. But for filamentous fungi, how effector proteins are transported into host cells remains unknown, although specialized infection structures such as haustoria are involved in the transport of effector proteins. Oomycete effector proteins ...

AI Technical Summary

Problems solved by technology

The transport of Magnaporthe oryzae effector proteins in rice root tissues has been reported, but there is no report on the transport of Magnaporthe oryzae proteins in rice plants, that is, from rice root tips to leaf buds