Is it Possible or not to Obtain Virus-free Bulblets of Lilium sp by Meristem Culture?

（1）迪巴法（DIBA）是检测百合花叶病毒（LSV）、郁金香花叶病毒（TBV-L）和黄瓜花叶病毒（CMV）的一种简便、可靠的方法。(2)In对18个百合品种进行迪巴检测，约70%的百合检测到LSV，部分百合还检测到TBV-L、CMV或两者共感染。LSV感染的百合植株生长不受影响，而LSV与TBV-L、CMV或三种病毒感染的百合植株叶片卷曲、花叶或花形、花色异常。（3）三种病毒感染母种鳞茎后，除“卡萨布兰卡”和美丽百合的三种病毒在鳞片培养的小鳞茎中约60%未检出外，其余几种病毒在鳞片培养的小鳞茎中均检出。（4）将小鳞茎原基的分生组织和鳞片培养的小鳞茎原基的分生组织（简化的分生组织培养）进行分离培养，前者70%以上的分生组织发育成小鳞茎，后者80%以上的分生组织发育成小鳞茎。（5）用酶免疫组织化学方法检测了LSV在鳞片培养的小鳞茎和小鳞茎原基的分生组织顶端的分布：小鳞茎原基及其周围组织中未检测到LSV，而在第二叶原基和第一叶原基基部中检测到LSV，但在分生组织顶端未检测到LSV。（6）两种方法都能获得大量的脱毒小鳞茎，但脱毒率因品种和病毒种类而异。（7）接种LSV和CMV的小鳞茎在35 ℃下培养10天以上，未检测到CMV，但LSV仍有残留。在35 ℃下培养的小鳞茎的分生组织比在25 ℃下培养的小鳞茎的分生组织的脱毒率高。

(1)DIBA (dot-immunobinding assay) was found to be a simple, reliable method to detect LSV (lily symptomless virus), TBV-L (tulip breaking virus-lily), and CMV (cucumber mosaic virus) as well as indirect ELISA (enzyme-linked immunosorbent assay). (2)In the DIBA testing of 18 lily varieties growing in the field, LSV was detected in ca.70% lilies, and co-infection with TBV-L,CMV or both were also detected in several lilies. Lily plants infected with LSV were symptomless in their growth but those with LSV and TBV-L or CMV or with three viruses had curling leaves, mosaic leaves or abnormal flowers in shape and color. (3)When mother bulbs were infected with three viruses, those viruses were detected in almost all bulblets grown from the scale cultures, except for those of'Casablanca'and L.speciosum in which three viruse were not detected in ca.60% bulblets grown from the scale cultures. (4)Meristem excised from bulblets and those from bulblet primordia developing adventitiously on scale cultures (a simplified meristem culture) were cultured to obtain virus-free bulblets ; more than 70% of the meristems excised in the former method and more than 80% of them in the latter one developed into bulblets. (5)The distribution of LSV in the meristem tip of bulblets and bulblet primordia on scale cultures was examined by enzyme-immunohistochemistry : no LSV was detected in the bulblet primordium and the surrounding tissue ; and LSV was observed in the second leaf primordium and the base of the first one but not in the meristem tip. (6)many virus-free bulblets were obtained by two methods, but the percentages depended on the varieties of lily examined and the types of viruses. (7)When bulblets infected with LSV and CMV were incubated at 35ﾟC for more than 10 days, CMV was not detected in the bulblets, but LSV remained. Meristems excised from the bulblets incubated at 35ﾟC yielded virus-free bulblets at higher rate compared with those excised from bulblets cultured at 25ﾟC.