【作者】 马翠萍；

【导师】 孙仲序；

【作者基本信息】 山东农业大学 ， 果树学， 2002， 硕士

【摘要】 本试验于2000～2002年在山东农业大学和泰安山口格林农业生物技术有限公司进行。试验试材主要有果树(樱桃colt砧木、草莓、冬枣、石榴)、花卉(月季、一品红、蔷薇)、林木(八里庄杨、速生杨、日本构树)。针对当前植物组织培养中污染率过高，在科研和生产中造成人力、物力和财力巨大浪费的现状，对植物组织培养污染进行了系统的研究。主要研究结果如下： 1．根据微生物的菌落特征、光学显微镜观察、昆虫的形态特征和生活习性进行了污染物种类的鉴定，认为污染物主要有真菌(青霉菌、镰刀菌、黑曲霉)、细菌(革兰氏阳性和革兰氏阴性)和昆虫(螨和蓟马)。 2．在植物组织培养条件下，以菌落直径为指标，测定了不同真菌的繁殖生长进程。利用分光光度计采用比浊法测定了细菌的繁殖生长状况。从真菌和细菌的生长曲线可以看出，两者都有一个快速生长阶段。 3．对培养瓶和外植体各部位的菌类分布，做了系统调查，试验结果表明：外植体各部位的菌类分布有一定的差异，新叶和芽带菌较少；培养瓶的瓶壁和封口纸都能携带污染菌，成为污染源之一。 4．对培养室和接种室采用紫外线和臭氧发生器两种灭菌方法进行灭菌比较试验，认为臭氧比紫外灯方式杀菌能力强，消毒效果好。 5．试验证明植物导管直径大于细菌100倍以上，细菌完全能够通过外植体导管进入到外植体内部，成为外植体的内生菌，引起接种后的组培苗污染。 6．提出植物组培苗污染防治的四项措施：第一，在温室中采的外植体接种成活率高于大田中采的外植体，同时对大田中的外植体亲本进行预处理也能提高接种成活率。第二，对组培苗要勤检查，及时挖除污染培养基和转移未污染的外植体。第三，在培养基中加入防腐剂以及抗生素和医用杀菌剂混合物，进行综合防治。第四，已污染的组培苗采用滤纸桥瓶外生根法挽救。更多还原

【Abstract】 Studies on the contamination of plant tissue culture was studied in Shandong Agricultural University and Tai’an Agricultural Science and Technology Company during 2000-2002,using some kinds of fruit trees (P sernlata Ledeb; C erasiis vulgavis MilL Fradaria vesca L^ Ziziphus jujube mill; Z saliva Gaevtn^ Punica granat um L.),flowes (Rosa rugosa^ Euphorbia pulcherrima^ R.multiflora) and forest trees (Balizhuang-yang^ Populus L. -. Broussonetiapapyrifera L.) as materials. The main results were as follows:1. According to colony character of the microbe, taking photographs by electron microscope, shape character, habit and characteristics of live of insect , the identification is made. As is thought that contamination was caused mainly by fungus (Fusarium > Aspergillus niger^ Penicillium ).bacterium(G^ and G~ ) and insect (acarus and thrips ).2. Under the condition of plant tissue culture, having the diameter of fungus community as guide line, the propagation and growth of different fungus is measured. Status of propagation and growth of bacterial is measured by spectrophotometer. From the growth status of fungus and bacteria, a rapid growth phase could be seen in both of them.3. The distributing of fungoid both explant and culture bottle was investigated in system, the result of the experiment showed that there was some difference in the distributing of fungoid between every part of explant, and there was less fungoid in the newly born leaves and buds; The wall of the culture bottle and the seal paper could also carry fungoid which brought about contamination of plant tissue culture, and both of them became one of the fountainhead of contamination.4. The fungoid on the culture room and inoculation room was extinguished by using two kinds of method including ultraviolet radiation and ozonizer. As was thought that it was better to extinguish the fungoid using ozonizer than ultraviolet radiation, the effect of ozonizer was better and it could thoroughly extinguish the fungoid without leaving any dead angle.5. When the diameter of the plant trachea was 100 times bigger than that of bacteria,the experiment showed that bacterial could thoroughly enter into the inside of the explant from the outside of the explant and turned into the endogenesis fungus, then that caused the contamination of the plantlet after the inoculation.6. Four steps was summed up on the prevention and cure about the contamination of plant tissue culture: First, the material used as inoculation should be collected in spring and autumn when the propagation of fungoid was slow; sprouting inside the room by collecting rigid branch outside the room in winter, then inoculating with young buds; avoiding as possible inoculation in summer, and spraying a certain concentration antiseptic medicine before collecting the material from the female parent. Second, check-up of the plantlet should be made frequently, and the contaminated medium should be excavated and the explant uncontaminated should be transferred in time. Third, integration of prevention and cure could be made by adding antibiotic and antiseptic used in medicine; Fourth, the plantlet having been contaminated could be saved by rooting in filter-paper bridge outside the bottle.更多还原