[1]杨雅运,傅华君,苏佳露,等.竹叶去硅方法的筛选及其细胞学研究[J].南京林业大学学报(自然科学版),2019,43(05):074-80.[doi:10.3969/j.issn.1000-2006.201903003]
 YANG Yayun,FU Huajun,SU Jialu,et al.Desilication of bamboo leaves for paraffin slices preparation and their cytological study on eleven bamboo species[J].Journal of Nanjing Forestry University(Natural Science Edition),2019,43(05):074-80.[doi:10.3969/j.issn.1000-2006.201903003]
点击复制

竹叶去硅方法的筛选及其细胞学研究
分享到:

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
43
期数:
2019年05期
页码:
074-80
栏目:
研究论文
出版日期:
2019-09-20

文章信息/Info

Title:
Desilication of bamboo leaves for paraffin slices preparation and their cytological study on eleven bamboo species
文章编号:
1000-2006(2019)05-0074-08
作者:
杨雅运12傅华君12苏佳露12万雅雯 12张春霞 1丁雨龙1林树燕1*
(1.南京林业大学,南方现代林业协同创新中心,南京林业大学竹类研究所,江苏 南京 210037; 2.南京林业大学生物与环境学院,江苏 南京 210037)
Author(s):
YANG Yayun12 FU Huajun12 SU Jialu12 WAN Yawen12 ZHANG Chunxia1DING Yulong1 LIN Shuyan1*
(1.Co-Innovation Center for the Sustainable Forestry in Southern China, Bamboo Research Institute, Nanjing Forestry University, Nanjing 210037, China; 2. College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, China)
关键词:
竹叶 石蜡切片 去硅 解剖结构
Keywords:
bamboo leaves paraffin section desiliconization anatomical structure
分类号:
S795
DOI:
10.3969/j.issn.1000-2006.201903003
文献标志码:
A
摘要:
【目的】竹类植物是我国重要的经济植物资源,竹叶表皮细胞富含硅质,导致制片过程中切片易破碎,不能获得清晰完整的竹叶切片。开发竹叶切片处理方法,对于研究竹类植物叶片解剖结构具有重要的实践和应用价值。筛选有效的氢氟酸处理浓度以及时间是观察竹叶内部解剖结构的前提。【方法】采用石蜡制片的方法,以竹叶为材料,首先优化去硅环节,筛选出适用于竹叶的最佳氢氟酸浓度和时间; 之后采用此最佳处理方法制片,观察竹叶内部细胞结构并结合去硅效果进行分析。【结果】通过对竹叶叶表皮和基本组织的形态解剖结构进行比较,发现真正影响竹叶完整石蜡切片制备的结构是叶片上下表皮的硅质细胞和主脉,这些结构使得制片易破碎,无法得到完整竹叶内部解剖结构。通过试验发现经过体积分数为25%的氢氟酸去硅处理2 d后再进行石蜡制片可以使竹叶上下表皮细胞和叶脉结构完整,得到理想的石蜡切片。【结论】竹叶去硅处理最佳氢氟酸体积分数为25%,处理时间以2 d为宜。
Abstract:
【Objective】 Bamboo is an important economic plant in China. As the epidermis cells of bamboo leaves are rich in silicon, paraffin slices of bamboo epidermal cells are easily broken and thus rarely provide a clear and complete sample. Nevertheless, the paraffin method for preparation of bamboo leaf sample has important practical applications in the study of bamboo leaf anatomical structure. Therefore, it is very necessary to desiliconize with hydrofluoric acid before paraffin sectioning. Determining the optimum concentration of hydrofluoric acid for treatment and treatment duration is crucial for observing the internal anatomical structure of bamboo leaves. 【Method】 In this study, we used the paraffin method to prepare leaf samples of 11 bamboo species. First, we determined the optimum hydrofluoric acid concentration and treatment duration for the desilication of bamboo leaves; the other 10 bamboo species were then subjected to this treatment. The bamboo leaf cell structure as well as the effect of desilication were observed and analyzed. 【Result】 By comparing the morphological and anatomical structures of the leaf epidermis with those of leaf basic tissues, the structures that markedly affected the quality of the paraffin slice were determined to be the siliceous cells of the upper and lower epidermis and the main vein. These structures render the traditional paraffin slice easily breakable and make it difficult to obtain a paraffin slice of the whole internal anatomical structure of bamboo leaves. In this experiment, we found that with desilication of the upper and lower epidermal cells and veins of bamboo leaves using 25% hydrofluoric acid for 2 days resulted in the successful preparation of complete paraffin sections. 【Conclusion】 The optimum hydrofluoric acid concentration for desilication is 25% and the treatment duration is 2 days.

参考文献/References:

[1] 郑钧,吴仁武,史琰,等. 竹类植物的主要环境效应研究进展[J]. 浙江农林大学学报, 2017, 34(2): 374-380. DOI: 10.11833/j.issn.2095-0756.2017.02.024. ZHENG J, WU R W, SHI Y, et al. Research progress on environmental effects of bamboo: a review [J]. Journal of Zhejiang A&F University, 2017, 34(2): 374-380. [2] 李岚,朱霖,朱平. 中国竹资源及竹产业发展现状分析[J]. 南方农业, 2017, 11(1): 6-9. DOI: 10.19415/j.cnki.1673-890x.2017.1.002. LI L, ZHU L, ZHU P. Analysis on the present situation of bamboo resources and bamboo industry in China[J]. South China Agriculture, 2017, 11(1): 6-9. [3] 杨丽,娄永峰,彭镇华,等. 毛竹、麻竹光合途径类型分析[J]. 南京林业大学学报(自然科学版), 2015, 39(5): 169-173. DOI: 10.3969 /j.issn.1000-2006.2015.05.028. YANG L, LOU Y F, PENG Z H, et al. A preliminary study on the type of photosynthetic pathway in Phyllostachys edulis and Dendrocalamus latiflorus [J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2015, 39(5): 169-173. [4] 陈晓亚,胡成华. 方竹属(竹亚科)叶片表皮微形态特征[J]. 植物分类学报, 1993, 31(3): 227-235. CHEN X Y, HU C H. Micromorphological characteristics of leaf epidermis of Chimonobambusa(Bambusoideae)[J]. Journal of Plant Classification, 1993, 31(3): 227-235. [5] 丁雨龙,赵奇僧,陈志银,等. 竹叶结构的比较解剖及其对系统分类意义的评价[J]. 南京林业大学学报, 1994, 18(3): 1-6.DOI:10.3969/j.issn.1000-2006.1994.03.01. DING Y L, ZHAO Q S, CHEN Z Y, et al. Comparative anatomy of bamboo leaf structure and its evaluation on systematic classification [J]. Journal of Nanjing Forestry University, 1994, 18(3): 1-6. [6] 白琼,于丽霞,鄢波. 竹类植物分类研究进展[J]. 河南科技, 2014, 5(9): 188-189. BAI Q, YU L X, YAN B. Research advances in bamboo taxonomy [J]. Journal of Henan Science and Technology, 2014, 5(9): 188-189. [7] 朱振贤,张芬耀,宋盛,等. 竹亚科植物分类研究进展[J]. 世界林业研究, 2017, 30(3): 35-40. DOI: 10.13348/j.cnki. sjlyyj.2017.0033.y. ZHU Z X, ZHANG F Y, SONG S, et al. Research advances in bamboo taxonomy [J]. World Forestry Research, 2017, 30(3): 35-40. [8] 丁雨龙. 刚竹属(Phyllostachys)系统分类的研究[D]. 南京:南京林业大学, 1998. DING Y L. Studies on the systematic taxonomy of the genus Phyllostachys [D]. Nanjing: Nanjing Forestry University, 1998. [9] 钱领元,方伟. 国产16种竹叶的比较解剖研究[J]. 竹子研究汇刊, 1986, 5(2): 78-86. QIAN L Y, FANG W. Comparative anatomy of 16 species of bamboo leaves in China[J]. Journal of Bamboo Research, 1986, 5(2): 78-86. [10] 徐德克,吕厚远,李泉. 竹亚科植硅体形态学研究及其生态学意义[J]. 第四纪研究, 2005, 25(6): 777-784. XU D K, LÜ H Y, LI Q. Morphological study of phytolith in phyllostachys pubescens and its ecological significance [J]. Quaternary Sciences, 2005, 25(6): 777-784. [11] ZHANG C, WANG L, ZHANG W, et al. Do lignification and silicification of the cell wall precede silicon deposition in the silica cell of the rice(Oryza sativa L.)leaf epidermis? [J]. Plant and Soil, 2013, 372(1/2): 137-149. DOI: 10.1007/s11104-013-1723-z. [12] 何蕊,邱坚,罗蓓,等. 慈竹植硅体形态及其发育变化研究[J]. 西北农林科技大学学报(自然科学版), 2018, 46(4): 68-74. DOI: 10.13207/j.cnki.jnwafu.2018.04.008. HE R, QIU J, LUO B, et al. Developmental changes and morphology of phytolith in Bambusa emeiensis [J]. Journal of Northwest A&F University(Natural Science Edition), 2018, 46(4): 68-74. [13] 曹尚银,陈利娜,薛辉,等. 石榴花芽石蜡切片制作方法的改良[J]. 安徽农业科学, 2017, 45(2): 1-3. DOI: 10.13989/j.cnki.0517-6611.2017.02.001. CAO S Y, CHEN L N, XUE H, et al. Improvements of traditional paraffin section for pomegranate floral bud [J]. Journal of Anhui Agricultural Science, 2017, 45(2): 1-3. [14] 龙春玲,刘腾飞,于芬,等. 厚壁毛竹与毛竹叶片的光学解剖结构比较分析[J]. 安徽农业大学学报, 2015, 42(1): 39-44. DOI: 10.13610/j.cnki.1672-352x.20141225.011. LONG C L, LIU T F, YU F, et al. Comparative anatomy of leaves between Phyllostachys edulis ‘Pachyloen' and Phyllostachys edulis [J]. Journal of Anhui Agricultural University, 2015, 42(1): 39-44. [15] 蔡联炳,郭延平. 禾本科植物叶片表皮结构细胞主要类型的演化与系统分类和发育途径的探讨(续)[J]. 西北植物学报, 1996, 16(1): 65-72. CAI L B, GUO Y P. Studies on constituent cells of leaf epidermis, systematics and phylogenetic path of the family Poaceae(continued)[J]. Journal of Northwest Botany, 1996, 16(1): 65-72. [16] 尤华明. 4种竹叶超薄切片技术的研究[J]. 福建林学院学报, 2000, 20(1): 31-34. DOI: 10.13207/j.cnki.jnwafu.2018.04.008. YOU H M. Study on four kinds of ultra-thin section techniques of bamboo leaves [J]. Journal of Fujian Forestry University, 2000, 20(1): 31-34. [17] 魏欣悦,苏源,杨静,等. 微波快速石蜡切片法观察水稻叶片组织[J]. 云南农业大学学报(自然科学版), 2011, 26(4): 454-457. DOI: 10.3969/j.issn.1004-390X(n).2011.04.004. WEI X Y, SU Y, YANG J, et al. Microwave paraffin section in rice leaf tissue [J]. Journal of Yunnan Agricultural University(Natural Science Edition), 2011, 26(4): 454-457. [18] 苏佳露,林树燕,史无双,等. 6个竹种竹叶的解剖形态观察与三维构建[J]. 南京林业大学学报(自然科学版), [2019-09-16]. http://kns.cnki.net/kems/detail/32.1161.S.20190404.1015.002.html. DOI: 10.3969/j.issn.1000-2006.201810044. SHU J L, LIN S Y, SHI W S, et al. Anatomical observation and three-dimensional construction of leaf blades from six bamboos [J]. Journal of Nanjing Forestry University(Natural Sciences Edition), [2019-09-16]. http://kns.cnki.net/kems/detail/32.1161.S.20190404.1015.002.html. [19] VIEIRA R C, GOMES D M, SARAHYBA L S, et al. Leaf anatomy of three herbaceous bamboo species [J]. Braz J Biol, 2002, 62(4B): 907-922. [20] 刘济明,李丽霞,赵晓鹏,等. 小蓬竹不同天然居群解剖形态差异[J]. 江苏农业科学, 2016, 23(5): 231-233. DOI: 10.15889/j.Issn.1002-1302.2016.05.066. LIU J M, LI L X, ZHAO X P, et al. Differences in anatomical morphology of different natural populations of Drepanostachyum luodianense [J]. Jiangsu Agricultural Science, 2016, 23(5): 231-233.

相似文献/References:

[1]马自超,吴伟志,彭洪斌,等.由竹叶制取叶绿素铜钠盐的研究[J].南京林业大学学报(自然科学版),1991,15(01):064.[doi:10.3969/j.jssn.1000-2006.1991.01.012]
 Ma Zichao Wu Weizhi Chen YiyuenPeng Hongbin Liao Yiming Zhuan Quanhuo(Nanjing Forestry University).A STUDY ON THE PREPARATION OF THE SODIUM COPPER CHLOROPHYLLIN FROM BAMBOO LEAVES[J].Journal of Nanjing Forestry University(Natural Science Edition),1991,15(05):064.[doi:10.3969/j.jssn.1000-2006.1991.01.012]
[2]丁雨龙,赵奇僧,陈志银,等.竹叶结构的比较解剖及其对系统分类意义的评价[J].南京林业大学学报(自然科学版),1994,18(03):001.[doi:10.3969/j.jssn.1000-2006.1994.03.001]
 Ding Yulong,Zhao Qiseng(Nanjing Forestry Forestry University Nanjing 007)Cheng Zhiyin,Wang Quansheng(Forestry Bureau of Jiangsu Province).STUDIES ON THE COMPARATIVE ANATOMY OF BAMBOO LEAVES AND ITS SIGNIFICANCE FOR BAMBOO SYSTEMATIC TAXONOMY[J].Journal of Nanjing Forestry University(Natural Science Edition),1994,18(05):001.[doi:10.3969/j.jssn.1000-2006.1994.03.001]

备注/Memo

备注/Memo:
收稿日期:2019-03-01 修回日期:2019-05-07 基金项目:“十二五”国家科技支撑计划(2015BAD04B0305); 农业科技自主创新资金项目(CX(16)1005-2-2); 科技支撑计划(农业)重点项目(BE2016304); 江苏高校优势学科建设工程资助项目(PAPD)。 第一作者:杨雅运(1036654249@qq.com)。*通信作者:林树燕(lrx@njfu.com.cn),教授,博士,ORCID(0000-0001-6326-2516)。 引文格式:杨雅运,傅华君,苏佳露,等. 竹叶去硅方法的筛选及其细胞学研究[J]. 南京林业大学学报(自然科学版),2019,43(5):74-80.
更新日期/Last Update: 2019-10-08