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Immunological Detection and Localization of CLSV -infected Fuji Apple by Tissue Blotting

Wang Qiaochun(Research Centre for Horticulture Plants. Sichuan Academy of Agricultural Sciences, Chengdu 610066)  
Shoot-tip explants of Fujiapple were established and micropropagated on MS medium. Callus and mi-croshoot with 4 6 weeks old were used for the present study. Sections(lmm)were cut from fresh tissuesby hand using razor blade,Tissue printings were made by pressing the freshly cut tissue surface on nitrocel-lulose membrances. After tissue blottings were blocked,CLSV antigen in tissue blottings were reacted withCISV-specific alkaline-phosphatase-labelled antibody. Then, the reacting results were coloured. The posi-tive reaction on tissue blottings showed a purple colour, while the negetive one produced no colour.Resultsshowed that CLSV in both shoots and calli was readily detected on tissue bolttings. Colour reaction of CLSV-infected callus was more sensitive than that of CLSV-infected shoot.There exsited two types of CLSV distri-bution in callus,a throughout and an uneven distribution. CLSV distribution in shoot was found in threetypes.Firstly, CLSV was located in epidermis,cortex and Vascular svstem, but never in pith.Secondly,CLSV was unevenly distributed with a main concentration in epidermis and only a quite few in cortex andphloem. Thirdly, CLSV distribution was like a" chimerd",i.e. a part of the same tissue was CLSV-infectedand the other were healthy, The suecessful development of tissue blotting technique for CLSV detection andlocalization opened a new way for studies on fruit tree viruses.
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