摘要
以花果发育周期2年的麻栎(Quercus acutissima)为试验材料,测试不同组织制片软化方案,摸索适合石蜡切片显微制片方法,并对不同软化处理后的材料进行切片和花粉管荧光染色,以确定对下游试验的影响。结果表明,对于雌花发育早期,花柱及子房壁木质化程度较低的材料,采用1.0 mol/L NaOH、1.0 mol/L HCl和10%Na2SO3高温高压软化各1.5 h,均可在1 d内完成软化及解剖,但对木质化程度高的雌花材料效果不佳。甘油乙醇软化剂、冰醋酸软化2种方法软化木质化程度高的雌花材料有一定软化效果,但耗时较长,处理需14~180 d。采用5%乙二胺溶液,40℃处理对高木质化雌花进行软化效果最佳,仅需2~4 d即可达到预期。经乙二胺软化后材料褐化明显,需要进一步漂白,对下游组织染色有一定影响,但仍为高木质化雌花软化的最佳处理方法。该研究为高木质化花材料的显微制片软化方案提供重要解决方案。
Quercus acutissima,a biannual fruiting species was used to optimize the tissue softening protocols that were suitable for paraffin section.Moreover,the pollen tubes using Aniline Blue were stained to assess the possible side-effects of the softening to the downstream experiments.The results showed that the softening on the early developing stage pistillate with lignification could be well achieved using 1.0 mol/L NaOH or 1.0 mol/L HCl or using 10%Na2SO3 to autoclave for 1.5 hours,respectively,but these methods did not work well on the highly lignified pistillate flowers at later developing stage.The traditional glycerol-ethanol softening and glacial acetic acid soften methods took a long time(more than 14 days up to 180 days)to soft the flower,which was unpractical.Using 5%ethylenediamine solution at 40℃ could efficiently soften the high-lignified pistillate female flowers within 2-4 days,which was the best time-cost efficient method to soften the high lignified flowers.However,this protocol would cause severe tissue browning,which required further whitening treatment using 0.3%bleach.This paper provided an comprehensive solution to proceed the microtome study on highly lignified flowers at different developing stage.
作者
史成成
宁馨
李颖
曹建国
SHI Cheng-cheng;NING Xin;LI Ying(College of Life Sciences,Shanghai Normal University,Shanghai 200234;Shanghai Chenshan Plant Science Research Center,Chinese Academy of Sciences,Shanghai Chenshan Botanical Garden,Shanghai 201602;The Laboratory of Forestry Genetics,Central South University of Forestry and Technology,Changsha,Hunan 410004)
出处
《安徽农业科学》
CAS
2020年第5期4-8,共5页
Journal of Anhui Agricultural Sciences
基金
上海市绿化和市容管理局科技攻关项目(G162404)
关键词
栎属
高度木质化雌花
组织软化方法
显微制片
Quercus
Lignified pistillate flowers
Tissue softening protocol
Microtome
作者简介
史成成(1994-),女,山东德州人,硕士研究生,研究方向:植物受精生物学。
