用CEV直接感染柑桔愈伤组织的接种方法的研究

STUDIES ON INOCULATING METHODS OF DIRECT INFECTION OF CITRUS CALLI WITH CEV

根据ＣＥＶ机械传染及高温诱导发病的特性，用三种方法直接接种柑桔愈伤组织，结合形态学观察、增殖率比较和电泳检测，结果表明，带毒切伤浸泡是ＣＥＶ离体感染柑桔愈伤组织的有效方法。３０℃培养条件下，感染ＣＥＶ的愈伤组织大约在８ｄ后发生明显褐变，并会逐渐加深，同时，感染后的愈伤组织的质地也趋坚硬，且表现出特殊的病理生长特性，前期（０—２０ｄ）显著地较健康对照生长快，后期（２０—４０ｄ）则明显减缓甚至停止生长。

Three methods of direct inoculation of in vitro citrus calli with CEV weredescribed in this paper, based on its characteristics that CEV can be mechanically transmi-tted and that its pathogenesis can be stimulated by relative high temperature. The resultsshowed that inoculation through cutting and soaking with CEV-RNA extracts was the mostefficient method of in vitro CEV-infection of citrus calli, according to morphologicalobservation, comparison of the rate of increase(RI) and two-dimensional electrophoreticanalysis of inoculated calli. Compared with control(both cultured in 30℃), CEV-infectedcalli grew brown evidently from 8 days after inoculation, and their texture was gettingstiff. At the same time, infected calli had special pathological rate of increase that theygrew more quickly in early phase(0 20 days)and became much slower and almost stoppedincreasing is subcultural period(20 40 days).