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

(1)斑点免疫结合法是检测百合无症状病毒、郁金香破裂病毒L和黄瓜花叶病毒的简便、可靠的方法，也是检测黄瓜花叶病毒的间接酶联免疫吸附试验。(2)对大田种植的18个百合品种进行的DIBA检测中，约有70%的百合品种检测到LSV，也有几个百合品种检测到与黄瓜花叶病毒L、黄瓜花叶病毒的混合感染。侵染LSV的百合植株生长无症状，而LSV、烟草花叶病毒L、黄瓜花叶病毒及3种病毒的百合植株均出现卷叶、花叶或花色异常。(3)用3种病毒侵染鳞茎时，除‘Casablanca’和L.Speciosum的鳞茎外，几乎所有鳞茎都检测到这3种病毒，约60%的鳞茎未检测到3种病毒。(4)将鳞茎切取的分生组织和鳞茎原基的不定芽(简化分生组织培养)培养成脱毒小鳞茎，前者切取的分生组织70%以上，后者切取的分生组织中80%以上发育成小鳞茎。(5)用酶免疫组织化学方法检测了鳞茎培养小鳞茎和小鳞茎原基分生组织顶端的LSV分布：小鳞茎原基及其周围组织未检测到LSV，第二叶原基和第一叶原基的根部未检测到LSV，分生组织顶端未检测到LSV。(6)两种方法均可获得较多的脱毒小鳞茎，但脱毒百分率与百合品种和病毒种类有关。(7)在35ﾟ℃下培养10天以上，未检测到黄瓜花叶病毒，但黄瓜花叶病毒仍存在。与在25ﾟC培养的小鳞茎相比，从35ﾟC培养的小鳞茎中分离出的小鳞茎产生脱毒小鳞茎的比率更高。

(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.