摘要
为对红松胚性愈伤组织诱导过程进行组织细胞学观察,探索针对带有愈伤组织的红松种子的石蜡切片制作方法,本研究以带有愈伤组织的红松未成熟种子为实验材料,在传统石蜡切片方法的基础上,针对制片方法、脱水、粘片、染色等重要环节进行改良。研究结果表明,片染方法明显好于整染方法,可以保证组织切片的完整性;此外,筛选出适宜的乙醇脱水时间,优化了染色方案,确定蛋白-甘油粘片剂作为首选的粘片剂。优化后的石蜡切片方法有效地解决了含愈伤的红松种子切片愈伤组织破碎、细胞结构不清晰和蜡带脱落等问题,最终成功获得组织完整、细胞结构清晰的高质量切片。该方法可为红松胚性愈伤诱导过程中的解剖学研究提供技术参考。
In order to realize the histocytological observation of the embryogenic callus induction process of Pinus koraiensis,we explored the method of making paraffin sections for the seeds of P.koraiensis with calli.In this study,we used the immature seeds of P.koraiensis with calli as experimental materials,and improved some key links such as paraffin section making method,dehydration,sticking and dyeing etc on the basis of traditional paraffin section technique.The results demonstrated that the method of piece tissue dyeing was obviously better than the whole tissue dyeing method,which can ensure the integrity of the tissue section.In addition,we screened out the appropriate ethanol dehydration time,optimized the dyeing scheme,and determined protein-glycerin sticky tablet as the preferred sticky tablet.The optimized paraffin section method effectively solved the problems of broken calli,unclear cell structure,and wax band shedding in the paraffin sections of P.koraiensis seeds containing callus,and finally successfully obtained high-quality sections with intact tissue and clear cell structure.This method can provide technical reference for the anatomical research in the process of embryogenic callus induction of P.koraiensis.
作者
梁艳
刘德强
潘朋
张妍
唐美璇
Liang Yan;Liu Deqiang;Pan Peng;Zhang Yan;Tang Meixuan(School of Life Sciences and A&F,Qiqihar University,Qiqihar,161006)
出处
《分子植物育种》
CAS
北大核心
2022年第13期4455-4461,共7页
Molecular Plant Breeding
基金
国家自然科学基金项目(31800515
31772334)
齐齐哈尔市科技局项目(NYGG-201913)
黑龙江省教育厅基本业务专项(145109137)
大学生创新创业训练计划项目(202010232203)共同资助。
关键词
红松
种子
愈伤组织
石蜡切片
Pinus koraiensis
Seeds
Callus
Paraffin section
