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

是否有可能通过分生组织培养获得无病毒的百合球茎？

• 批准号: 06660030
• 负责人: NIIMI Yoshiji
• 金额: $ 1.15万
• 依托单位: Niigata university
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660030/
• 关键词: Lily virus; ELISA; DIBA; meristem culture; simplified meristem culture; enzyme-immunohistochemistry; thermotherapy; LSV; TBV-L; CMV; 染色液; ほ場栽培

(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.

(1)DIBA（点免疫结合试验）被发现是一种简单、可靠的检测LSV（百合无症状病毒）、TBV-L（郁金香破百合病毒）和CMV（黄瓜花叶病毒）以及间接ELISA（酶联免疫吸附试验）的方法。 (2)在对18个田间百合品种的DIBA检测中,约70%的百合检出LSV,部分百合中还检出TBV-L、CMV或两者混合感染。感染LSV的百合植株生长无症状，但感染LSV和TBV-L或CMV或感染三种病毒的百合植株出现卷叶、花叶或形状和颜色异常的花朵。 (3)当母球茎感染三种病毒时，几乎所有鳞片培养的鳞茎中均检出这三种病毒，但‘卡萨布兰卡’和L.speciosum除外，其中约60%的鳞片培养的鳞茎中未检测到这三种病毒。 (4)将鳞茎切下的分生组织与规模化培养中偶然发育的鳞茎原基(简化分生组织培养物)进行培养，获得脱毒鳞茎；前一种方法切除的分生组织70%以上，后一种方法切除的分生组织80%以上发育成鳞茎。 (5)酶免疫组化法检测鳞片培养物鳞茎分生组织尖端和鳞茎原基中LSV的分布：鳞茎原基及周围组织中未检测到LSV；在第二叶原基和第一叶原基基部观察到LSV，但在分生组织尖端未观察到LSV。 (6)两种方法均获得了较多的脱毒鳞茎，但脱毒比例取决于所检测的百合品种和病毒种类。 (7)当感染LSV和CMV的鳞茎在35℃培养10天以上时，鳞茎中检测不到CMV，但LSV仍然存在。与从25℃培养的鳞茎切下的分生组织相比，从35℃培养的鳞茎切下的分生组织以更高的比例产生无病毒鳞茎。