黄瓜果瘤石蜡切片制片技术

Paraffin Section Technology of Fruit Tumor Tissues in Cucumber(Cucumis sativus L.)

果瘤是黄瓜(Cucumis sativus L.)果实重要外观品质性状之一,与黄瓜的经济价值密切相关。为了获得黄瓜果实果瘤石蜡切片制作方法,研究其性状形态建成的细胞学机制,本研究以黄瓜有果瘤品种S52和无果瘤品种S42 (大刺)/S06 (小刺)为材料,在传统石蜡切片方法的基础上,对脱水、染色、脱蜡等重要环节进行改良。结果表明:脱水时增加乙醇浓度梯度,可缩短每级处理时间至30 min;染色用埃氏苏木精(Ehrlich’s Hematoxylin)整染法染色3 d,便于后续操作;最优切片厚度为8μm;脱蜡时经两次纯二甲苯后可显微镜检精选,缩短获得高质量切片的时间。通过优化的制片方法可获得切片组织完整,结构清晰的高质量切片,是观察果瘤解剖学结构的有效方法。本研究为黄瓜果瘤切片技术提供一套完整、高效的操作步骤,为黄瓜其他器官及结构相似性状的解剖学研究提供技术参考。

Fruit tumor is one of the important appearance quality traits in cucumber(Cucumis sativus L.), which is closely related to the economic value of cucumber. In order to obtain the paraffin section technology and study the cytological mechanism of morphological formation for fruit tumor in cucumber, we used the samples of warty fruit line(S52) and non-warty fruit lines(S42 with big fruit spine and S06 with small fruit spine) as materials, and improved some key links such as dehydration, dyeing and dewaxing etc. on the basis of traditional paraffin section technology. Results demonstrated that increasing ethanol concentration gradient during dehydration can shorten the treatment time of each stage to 30 minutes. Samples were stained with Ehrlich’s hematoxylin for three days, which was convenient for subsequent operation. The optimal thickness of the section was 8 μm. After twice pure xylene dewaxing, sections could be examined and selected by microscope, which shortened the time for obtaining high-quality section. High quality sections with complete tissue and clear structure can be obtained by the optimized paraffin section technology, which is an effective method to observe the cytological structure of fruit tumor. This study provides a set of complete and efficient operation steps of paraffin section technology for cucumber fruit tumor, which will provide technical reference for anatomical research on other organs in cucumber and structural similar characteristics of other plants.