木本植物非均质化组织石蜡切片制作方法
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摘要
在常规石蜡切片技术的基础上,针对木本植物茎段木质化程度高、硬度大以及各部分组织硬度不均匀等特点,选取核桃(Juglans regia)茎段以及芽接愈合区域组织为实验材料,对固定、软化和脱水等关键步骤进行改进,获得结构完整且染色清晰的茎段组织和嫁接愈合区域组织切片,可清晰地观察到各部分组织的形态特征和愈合过程中的发育特征,且缩短了制片周期。采用改良后的实验流程成功获得了苹果(Malus pumila)、桃(Amygdalus persica)、杏(Armeniaca vulgaris)、李(Prunus salicina)和杨(Populus tomentosa)的茎段横截面切片。该方法为从解剖学上研究林木茎段生长机制和形态发育变化提供技术基础,为非均质化植物材料的石蜡切片制作提供参考。
In view of the characteristics of highly lignified, large hardness and heterogeneous of various parts of woody plants, the stem segments and healing areas of bud grafting of Juglans regia were used as materials with the conventional method of paraffin sectioning. The aspects of fixation, softening, dehydration and other critical steps were modified to obtain cross-section slices with complete organizational structure and clear staining. The morphological and developmental characteristics of the healing period could be observed clearly and shortened the experimental period. The paraffin sections of stem segments of apple, peach, apricot, plum and poplar were obtained by the improved experimental process. The study provides a technical basis for studying the growth mechanism and morphological development of stem segments of woody plants and provides a reference for making paraffin sections of heterogeneous plant materials.
In view of the characteristics of highly lignified, large hardness and heterogeneous of various parts of woody plants, the stem segments and healing areas of bud grafting of Juglans regia were used as materials with the conventional method of paraffin sectioning. The aspects of fixation, softening, dehydration and other critical steps were modified to obtain cross-section slices with complete organizational structure and clear staining. The morphological and developmental characteristics of the healing period could be observed clearly and shortened the experimental period. The paraffin sections of stem segments of apple, peach, apricot, plum and poplar were obtained by the improved experimental process. The study provides a technical basis for studying the growth mechanism and morphological development of stem segments of woody plants and provides a reference for making paraffin sections of heterogeneous plant materials.
引文
邓传远,郑俊鸣,张万超,郭素枝,薛秋华,叶露莹,孙建文(2015).红海榄木材结构的生态解剖.植物生态学报39,604-615.
丁平海,郗荣庭,李金空(1989).核桃愈伤组织形成条件及形成部位的解剖观察.河北农业大学学报12(1),27-33.
冯金玲(2011).油茶芽苗砧嫁接体愈合机理研究.博士论文.福州:福建农林大学.pp.22-28.
贺冰,李志岗,郝晓娟,贺运春(2014).植物材料快速石蜡制片方法.植物学报49,203-208.
侯春春,徐水(2009).浅析影响石蜡切片质量的关键因素.中国农学通报25(23),94-98.
黄坚钦,章滨森,陆建伟,付敢伟(2001).山核桃嫁接愈合过程的解剖学观察.浙江林学院学报18(2),111-114.
李和平(2009).植物显微技术(第2版).北京:科学出版社.pp.9-34.
李正理(1987).植物制片技术(第2版).北京:科学出版社.pp.138-148.
林月惠,李寒冰,贺新强(2001).高度木质化材料的冰冻切片技术.植物学通报18,118-120.
刘昊,宋晓波,周乃富,马庆国,裴东(2017).吲哚丁酸对核桃嫩枝扦插生根及内源激素变化的影响.浙江农林大学学报34,1038-1043.
刘杏娥,汪佑宏,江泽慧,费本华,任海青,田根林,徐鑫(2010).黄藤材发育过程中维管束的变化.西北林学院学报25(2),152-155.
鲁丹,张瑞,彭方仁,任莺,华宏(2013).红桤木扦插繁殖技术及生根过程的解剖学观察.西南林业大学学报33(2),24-28.
牛田,王厚新,李承秀,张林,于永畅,孙芳,王长宪(2012).华北五角枫嫩枝扦插生根形态解剖学分析.山东林业科技42(5),19-21.
权金娥,朱海兰,张春霞,张胜,赵忠(2014).四倍体刺槐茎段组织石蜡切片的制作方法.西北林学院学报29(3),140-144.
王欣,隋方功,初庆刚(2015).成熟玉米秆组织结构切片方法的比较研究.西北植物学报35,1052-1056.
吴华,陈娉婷,袁玲,徐永荣,陈龙清(2011).蕨类植物石蜡切片制作技术探讨.湖北农业科学50,3767-3769,3774.
吴清韩,赵庆芳,朱慧,马瑞君(2012).植物高度木质化叶片表皮细胞制备方法.植物学报47,422-426.
闫绍鹏(2011).欧美山杨杂种扦插生根的理化与分子机理研究.博士论文.哈尔滨:东北林业大学.pp.23-26.
杨虎彪,李晓霞,罗丽娟(2009).植物石蜡制片中透明和脱蜡技术的改良.植物学报44,230-235.
杨捷频(2006).常规石蜡切片方法的改良.生物学杂志23,45-46.
曾月星(1991).硬重木材的软化方法.木材工业5(2),45-46,32.
Coates PJ(1991).Paraffin section molecular biology:review of current techniques.J Histotechnol 14,263-269.
Day RC(2010).Laser microdissection of paraffin-embedded plant tissues for transcript profiling.In:Hennig L,K?hler C,eds.Plant Developmental Biology:Methods and Protocols.Totowa:Humana Press.pp.321-346.
Takahashi H,Kamakura H,Sato Y,Shiono K,Abiko T,Tsutsumi N,Nagamura Y,Nishizawa NK,Nakazono M(2010).A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection.JPlant Res 123,807-813.
丁平海,郗荣庭,李金空(1989).核桃愈伤组织形成条件及形成部位的解剖观察.河北农业大学学报12(1),27-33.
冯金玲(2011).油茶芽苗砧嫁接体愈合机理研究.博士论文.福州:福建农林大学.pp.22-28.
贺冰,李志岗,郝晓娟,贺运春(2014).植物材料快速石蜡制片方法.植物学报49,203-208.
侯春春,徐水(2009).浅析影响石蜡切片质量的关键因素.中国农学通报25(23),94-98.
黄坚钦,章滨森,陆建伟,付敢伟(2001).山核桃嫁接愈合过程的解剖学观察.浙江林学院学报18(2),111-114.
李和平(2009).植物显微技术(第2版).北京:科学出版社.pp.9-34.
李正理(1987).植物制片技术(第2版).北京:科学出版社.pp.138-148.
林月惠,李寒冰,贺新强(2001).高度木质化材料的冰冻切片技术.植物学通报18,118-120.
刘昊,宋晓波,周乃富,马庆国,裴东(2017).吲哚丁酸对核桃嫩枝扦插生根及内源激素变化的影响.浙江农林大学学报34,1038-1043.
刘杏娥,汪佑宏,江泽慧,费本华,任海青,田根林,徐鑫(2010).黄藤材发育过程中维管束的变化.西北林学院学报25(2),152-155.
鲁丹,张瑞,彭方仁,任莺,华宏(2013).红桤木扦插繁殖技术及生根过程的解剖学观察.西南林业大学学报33(2),24-28.
牛田,王厚新,李承秀,张林,于永畅,孙芳,王长宪(2012).华北五角枫嫩枝扦插生根形态解剖学分析.山东林业科技42(5),19-21.
权金娥,朱海兰,张春霞,张胜,赵忠(2014).四倍体刺槐茎段组织石蜡切片的制作方法.西北林学院学报29(3),140-144.
王欣,隋方功,初庆刚(2015).成熟玉米秆组织结构切片方法的比较研究.西北植物学报35,1052-1056.
吴华,陈娉婷,袁玲,徐永荣,陈龙清(2011).蕨类植物石蜡切片制作技术探讨.湖北农业科学50,3767-3769,3774.
吴清韩,赵庆芳,朱慧,马瑞君(2012).植物高度木质化叶片表皮细胞制备方法.植物学报47,422-426.
闫绍鹏(2011).欧美山杨杂种扦插生根的理化与分子机理研究.博士论文.哈尔滨:东北林业大学.pp.23-26.
杨虎彪,李晓霞,罗丽娟(2009).植物石蜡制片中透明和脱蜡技术的改良.植物学报44,230-235.
杨捷频(2006).常规石蜡切片方法的改良.生物学杂志23,45-46.
曾月星(1991).硬重木材的软化方法.木材工业5(2),45-46,32.
Coates PJ(1991).Paraffin section molecular biology:review of current techniques.J Histotechnol 14,263-269.
Day RC(2010).Laser microdissection of paraffin-embedded plant tissues for transcript profiling.In:Hennig L,K?hler C,eds.Plant Developmental Biology:Methods and Protocols.Totowa:Humana Press.pp.321-346.
Takahashi H,Kamakura H,Sato Y,Shiono K,Abiko T,Tsutsumi N,Nagamura Y,Nishizawa NK,Nakazono M(2010).A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection.JPlant Res 123,807-813.