白菜型油菜幼苗自发荧光及其克服的研究

Study on Autofluorescence of Brassica campestris Seedlings and Its Overcoming

本研究以洋葱表皮为对照植物,白菜型油菜为实验植物,研究洋葱表皮与白菜型油菜幼苗自发荧光现象,将黄皮洋葱和紫皮洋葱的内表皮和外表皮制作成临时装片观察其蓝、绿、红色三种自发荧光,研究观察到两种洋葱的内、外表皮均不显现三种荧光;活体材料观察白菜型油菜幼苗分为正常光照、黑暗处理和卡那霉素处理三种方式,三种处理方式的根、茎、叶组织均可观察到蓝色荧光,但光照条件下生长的叶与茎组织会出现红色荧光覆盖蓝色或绿色荧光的现象;三种处理方式的根均可发出微弱的绿色荧光,黑暗中水培法处理的油菜幼苗茎和叶片组织的绿色荧光强于根组织,因荧光较弱,叶绿素的红色荧光常掩盖叶和茎组织的蓝色或绿色荧光;三种处理方式的油菜幼苗根、茎、叶组织均可观察到红色荧光,其中在光照条件下水培法处理的油菜幼苗根、茎、叶片组织的红色荧光强度最强,卡那霉素溶液处理的白化幼苗仅在150~200 μg/ml浓度下处理6 d时表现出能克服红色荧光对其他颜色荧光的干扰,黑暗处理的油菜幼苗根、茎、叶组织荧光最弱,将在光照培养条件下生长良好的油菜幼苗,浸泡在含有果胶酶(5%)、纤维素酶(5%)的0.1 mol/L的PBS溶液中浸泡处理24 h后,幼苗根、茎、叶细胞的绿色荧光消除,蓝色、红色荧光强度降低。

In this study, onion epidermis was used as the control plant and Brassica campestris was used as the experimental plant to study the autofluorescence phenomenon of onion epidermis and Brassica campestris seedlings. The inner and outer epidermises of yellow-epidermisned onion and purple-epidermisned onion were made into temporary sheets to observe their blue, green and red autofluorescence. It was observed that the inner and outer epidermises of the two onions did not show three kinds of fluorescence. Materials in vivo observation of Brassica campestris seedlings can be divided into three ways: normal light, dark treatment and kanamycin treatment. Blue fluorescence can be observed in roots, stems and leaves of the three ways, but red fluorescence covers blue or green fluorescence in leaves and stems grown under light conditions. The roots of the three treatments can emit weak green fluorescence. The green fluorescence of stems and leaves of Brassica campestris seedlings treated by hydroponics in the dark is stronger than that of root tissues. Because of the weak fluorescence, the red fluorescence of chlorophyll often covers the blue or green fluorescence of leaves and stems. Red fluorescence can be observed in the roots, stems and leaves of Brassica campestris seedlings treated by three methods, and the red fluorescence intensity of roots, stems and leaves of Brassica campestris seedlings treated by hydroponics under light conditions is the strongest. The albino seedlings treated with kanamycin solution can only overcome the inter-ference of red fluorescence on other color fluorescence when treated with 150~200 μg/ml concentration for 6 days. The fluorescence of roots, stems and leaves of Brassica campestris seedlings treated in darkness is the weakest. When Brassica campestris seedlings that grow well in light cul-ture are soaked in 0.1 mol/L PBS solution containing pectinase (5%) and cellulase (5%) for 24 hours, the green fluorescence of roots, stems and leaves of the seedlings disappears, and the intensity of blue and red fluorescence decreases.