用点杂交方法定量检测植物病毒RNA负荷量

用RNA点杂交与用放射性成像仪直接读取放射性活度相结合的方法定量测定植物病毒RNA相对负荷量 ,检测了 0 0 0 1～ 1 0ppm浓度范围内二氢茉莉酸丙酯(PDJ)对烟草组织中烟草花叶病毒 (TMV)RNA的影响。结果表明 :在PDJ处理 3d时 ,与对照相比低浓度 ( 0 0 0 1ppm)PDJ处理的TMVRNA病毒负荷量无明显差异 ,但随着浓度的升高 ,PDJ对TMVRNA的复制和积累具有促进作用 ;浓度越高 ,促进作用越显著。同时用传统的枯斑接种法验证了上述结果。两种方法相比 ,点杂交结合放射性成像仪读取放射性活度的方法所受的人为误差干扰较小 ,能够相对精确地定量测定植物组织中病毒RNA负荷量。

QUANTITATIVE DETECTION FOR PLANT VIRUS’S RNA-LOADING BY DOT-BLOT

A new method,RNA dot blot combined with direct determination of the radioactivity by BIO Imaging Analyzer(dRH dBIA)was used for detecting RNA of plant virus in infected plant tissue.This method was used for the influence of RNA loading level of tobacco mosaic virus(TMV)in tobacco leave tissues after treatment of a plant hormone relatives(n Propyl dihydro jasmonate,PDJ)in the concentration range of 0 001～10ppm.The results indicate that after PDJ application onto tobacco leaves for 3days all PDJ treatments cause increase of TMV RNA loading level except 0 001ppm treatment,and the higher the concentration,the more obvious increase was observed..This phenomenon was confirmed with semi leaf lesion spot on Nicotiana glutinosa as a local lesion host.The dRH dBIA mehtod is applicable in quantitative determination of RNA without obvious artificatial influence.