Chinese Bulletin of Botany ›› 2018, Vol. 53 ›› Issue (5): 653-660.DOI: 10.11983/CBB18051
• TECHNIQUES AND METHODS • Previous Articles Next Articles
Zhou Naifu1, Zhang Junpei1,*(), Liu Hao2, Zha Weiwei1, Pei Dong1
Received:
2018-03-02
Online:
2018-09-01
Published:
2018-11-29
Contact:
Zhang Junpei
About author:
These authors contributed equally to this paper
Zhou Naifu, Zhang Junpei, Liu Hao, Zha Weiwei, Pei Dong. New Protocols for Paraffin Sections of Heterogeneous Tissues of Woody Plants[J]. Chinese Bulletin of Botany, 2018, 53(5): 653-660.
Figure 1 The intact cross-section slice of walnut stem segment tissue(A) Traditional FAA fixative, according to softening method 1 treatment; (B) Improve fixative, according to softening method 1 treatment; (C) Traditional FAA fixative, according to softening method 2 treatment; (D) Improve fixative, according to softening method 2 treatment. Bars=1 mm
Combination 1 | Combination 2 | Combination 3 | Combination 4 | |
---|---|---|---|---|
FAA fixative solution Softening method 1 | Improved fixative solution Softening method 1 | FAA fixative solution Softening method 2 | Improved fixative solution Softening method 2 | |
Paraffin section | Partial cavity, divided and ripples | Continuous and ripples | Continuous and tidy | Continuous and tidy |
Cortex | Brokenness and incomplete | Rupture and incomplete | Partial rupture and complete | Complete |
Pith | Partial cavity and removed | Partly dehiscent, not detached | Complete | Complete |
Transverse section | Fragmentary and incomplete with unclear outline | Shrinkage deformation, unclear outline | Partly complete with clear outline | Complete and smooth with clear outline |
Table 1 Effect of different types of fixation and softening on paraffin section of Juglans regia
Combination 1 | Combination 2 | Combination 3 | Combination 4 | |
---|---|---|---|---|
FAA fixative solution Softening method 1 | Improved fixative solution Softening method 1 | FAA fixative solution Softening method 2 | Improved fixative solution Softening method 2 | |
Paraffin section | Partial cavity, divided and ripples | Continuous and ripples | Continuous and tidy | Continuous and tidy |
Cortex | Brokenness and incomplete | Rupture and incomplete | Partial rupture and complete | Complete |
Pith | Partial cavity and removed | Partly dehiscent, not detached | Complete | Complete |
Transverse section | Fragmentary and incomplete with unclear outline | Shrinkage deformation, unclear outline | Partly complete with clear outline | Complete and smooth with clear outline |
Key steps of slicing | Traditional slicing method (method 1) | Improved slicing method (method 2) | |||
---|---|---|---|---|---|
Reagent | Time (h) | Reagent | Time (h) | ||
Fixation | FAA fixative | 48 | Improved fixative | 48 | |
Softening | 70% ethanol-glycerol | 288 | Hydrogen peroxide-glacial acetic acid, 70% ethanol-glycerol | 48 | |
Dehydration | Different concentrations of ethanol solution | 8.5 | Different concentrations of tert butanol solution | 6.5 | |
Transparency | Ethanol, xylene | 3 | - | - |
Table 2 Comparison of key steps of two different slicing methods
Key steps of slicing | Traditional slicing method (method 1) | Improved slicing method (method 2) | |||
---|---|---|---|---|---|
Reagent | Time (h) | Reagent | Time (h) | ||
Fixation | FAA fixative | 48 | Improved fixative | 48 | |
Softening | 70% ethanol-glycerol | 288 | Hydrogen peroxide-glacial acetic acid, 70% ethanol-glycerol | 48 | |
Dehydration | Different concentrations of ethanol solution | 8.5 | Different concentrations of tert butanol solution | 6.5 | |
Transparency | Ethanol, xylene | 3 | - | - |
Figure 2 The stem segment tissue slice of walnut with different treatment methods(A), (B) Traditional FAA fixative, according to softening method 1 treatment; (C), (D) Improve fixative, according to softening method 2 treatment. (B) and (D) are local amplification of (A) and (C). (A), (C) Bars=200 μm; (B), (D) Bars=100 μm. CZ: Cambium zone; Ph: Phloem; Xy: Xylem; Pi: Pith; PC: Parenchyma cell
Figure 3 The sliced section of walnut bud healing area using the improved paraffin section method(A)-(C) 6 days after grafted; (D)-(F) 12 days after grafted. (B), (C) and (E), (F) are local amplification of (A) and (D). (A), (D) Bars=1 mm; (B), (C), (E), (F) Bars=500 μm. CF: Callus formation; SC: Scion; ST: Stock; NL: Necrotic layer; DVB: Developing vascular bundles
Figure 4 The sliced sections of walnut, apple, peach, apricot, plum and poplar were obtained by improved paraffin section method(A)-(F) Paraffin sections of stems of walnut, apple, peach, apricot, plum and poplar; (G) and (H), (I) and (J), (K) and (L), (M) and (N), (O) and (P), (Q) and (R) are local amplification of (A)-(F). (A), (D), (E) Bars=1 mm; (G), (M), (O) Bars=500 μm; (H)-(L), (N), (P)-(R) Bars= 200 μm
1 | 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. |
2 | 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. |
3 | 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. |
4 | 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. |
5 | 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. |
6 | 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. |
7 | 李和平 (2009). 植物显微技术(第2版). 北京: 科学出版社. pp. 9-34. |
8 | 李正理 (1987). 植物制片技术(第2版). 北京: 科学出版社. pp. 138-148. |
9 | 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. |
10 | 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. |
11 | 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. |
12 | 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. |
13 | 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. |
14 | 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. |
15 | 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. |
16 | 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. |
17 | 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. |
18 | 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. |
19 | 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. |
20 | 杨捷频 (2006). 常规石蜡切片方法的改良. 生物学杂志 23, 45-46. |
21 | 曾月星 (1991). 硬重木材的软化方法. 木材工业 5(2), 45-46, 32. |
22 | Coates PJ (1991). Paraffin section molecular biology: review of current techniques.J Histotechnol 14, 263-269. |
23 | Day RC (2010). Laser microdissection of paraffin-embedded plant tissues for transcript profiling. In: Hennig L, Köhler C, eds. Plant Developmental Biology: Methods and Protocols. Totowa: Humana Press. pp. 321-346. |
24 | 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.J Plant Res 123, 807-813. |
[1] | Hailing Qi, Pengzhen Fan, Yuehua Wang, Jie Liu. Genetic diversity and population structure of Juglans regia from six provinces in northern China [J]. Biodiv Sci, 2023, 31(8): 23120-. |
[2] | Chun-Xia HE, Ping CHEN, Ping MENG, Jin-Song ZHANG, Hong-Guo YANG. Interspecific water use strategies of a Juglans regia and Isatis tinctoria/Senna tora agroforestry [J]. Chin J Plan Ecolo, 2016, 40(2): 151-164. |
[3] | Bing He, Zhigang Li, Xiaojuan Hao, Yunchun He. A New Technique of Fast Paraffin Sectioning in Plant Tissues [J]. Chinese Bulletin of Botany, 2014, 49(2): 203-208. |
[4] | ZHANG Cui-Ping, MENG Ping, LI Jian-Zhong, WAN Xian-Chong. Interactive effects of soil acidification and phosphorus deficiency on photosynthetic characteristics and growth in Juglans regia seedlings [J]. Chin J Plant Ecol, 2014, 38(12): 1345-1355. |
[5] | Jianxia Li, Chulan Zhang, Xiaofei Xia, Liangcheng Zhao. Cryo-sectioning Conditions and Histochemistry Comparison with Paraffin Sectioning [J]. Chinese Bulletin of Botany, 2013, 48(6): 643-650. |
[6] | ZHANG Jun-Zhou, GOU Xiao-Hua, ZHAO Zhi-Qian, LIU Wen-Huo, ZHANG Fen, CAO Zong-Ying, ZHOU Fei-Fei. Improved method of obtaining micro-core paraffin sections in dendroecological research [J]. Chin J Plant Ecol, 2013, 37(10): 972-977. |
[7] | Hubiao Yang;Xiaoxia Li;Lijuan Luo*. An Improved Clearing and De-waxing Method for Plant Paraffin Sectioning [J]. Chinese Bulletin of Botany, 2009, 44(02): 230-235. |
[8] | CHEN Liang-Hua, HU Ting-Xing, ZHANG Fan, LI Guo-He. GENETIC DIVERSITIES OF FOUR JUGLANSPOPULATIONS REVEALED BY AFLP IN SICHUAN PROVINCE, CHINA [J]. Chin J Plant Ecol, 2008, 32(6): 1362-1372. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||