竹叶去硅方法的筛选及其细胞学研究

doi:10.3969/j.issn.1000-2006.201903003

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南京林业大学学报（自然科学版） ›› 2019, Vol. 43 ›› Issue (5): 74-80.doi: 10.3969/j.issn.1000-2006.201903003

竹叶去硅方法的筛选及其细胞学研究

杨雅运¹^,²(), 傅华君¹^,², 苏佳露¹^,², 万雅雯¹^,², 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

收稿日期:2019-03-01 修回日期:2019-05-07 出版日期:2019-10-08 发布日期:2019-10-08
基金资助:
“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)

Desilication of bamboo leaves for paraffin slices preparation and their cytological study on eleven bamboo species

YANG Yayun¹^,²(), FU Huajun¹^,², SU Jialu¹^,², WAN Yawen¹^,², ZHANG Chunxia¹, DING 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

Received:2019-03-01 Revised:2019-05-07 Online:2019-10-08 Published:2019-10-08

摘要/Abstract

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。

关键词: 竹叶, 石蜡切片, 去硅, 解剖结构

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.

Key words: bamboo leaves, paraffin section, desiliconization, anatomical structure

中图分类号:

S795

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.

YANG Yayun, FU Huajun, SU Jialu, WAN Yawen, ZHANG Chunxia, DING Yulong, LIN Shuyan. 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(5): 74-80.DOI: 10.3969/j.issn.1000-2006.201903003.

图/表 4

图1

图2

图3

表1

11种竹的竹叶形态解剖特征比较"

竹种 bamboo species	竹叶类型 bamboo leaf type	厚度/μm thickness					截面积/μm² sectional area
竹种 bamboo species	竹叶类型 bamboo leaf type	叶片 blade	上表皮 upper epidermal	下表皮 lower epidermal	乳突 mastoid	叶肉细胞 mesophyll	泡状细胞 motor cell	主脉维管束鞘细胞 main bundle sheath	侧脉维管束鞘细胞 lateral bundle sheath
翠竹 Pleioblastus pygmaeus	功能叶	86.01±6.96^cde	10.81±1.95^cdefgh	9.31±1.15^ab	6.10±1.30^defgh	13.90±2.45^abcd	1 672.19±317.36^hi	128.85±48.73^def	88.85±29.85^defg
	嫩叶	77.89±6.56^f	11.6±1.27^bcde	10.2±1.59^a	5.38±1.09^fghij	11.25±1.97^efg	2 448.07±244.43^def	93.37±28.47^ef	87.37±23.66^dcef
	平均值	81.95±6.76	11.21±1.61	9.76±1.37	5.74±1.195	12.58±2.21	2 060.13±280.90	111.11±38.6	88.11±26.76
月月竹 Menstruocalamus sichuanensis	功能叶	103.59±9.76^a	10.37±2.64^defgh	7.95±0.92^bcdefg	7.19±1.61^def	14.35±1.91^abc	2 098.09±310.67^fgh	293.79±71.24^a	132.58±36.58^abc
	嫩叶	93.65±12.80^b	10±2.39^efghi	7.1±1.38^efgh	7.85±2.69^cd	11.21±1.97^efg	3 101.73±458.85^ab	248.63±29.95^efg	121.73±33.19^bcd
	平均值	98.62±11.28	10.19±2.52	7.53±1.15	7.52±2.15	12.78±1.94	2 599.91±384.76	271.21±50.6	127.16±34.89
乌哺鸡竹 Phyllostachys vivax	功能叶	86.42±4.70^f	7.56±1.13^j	6.89±1.03^fgh	17.37±4.09^a	13.59±2.99^abcd	1 710.76±220.79^ghi	250.36±16.09^ab	160.88±45.77^a
	嫩叶	78.16±3.26^cde	12.2±2.14^bcd	8.76±1.76^bcd	6.82±1.90^def	11.44±1.64^efg	2 427.51±406.97^def	244.63±38.85^ab	140.35±26.84^abc
	平均值	82.29±3.98	9.88±1.64	7.83±1.40	12.10±3.00	12.52±2.32	2 069.14±313.88	247.5±27.47	150.62±36.31
淡竹 Phyllostachys glauca	功能叶	78.11±4.18^f	9.75±2.09^efghi	7.29±0.91^defgh	7.30±1.39^de	13.52±1.51^bcd	1 681.61±302.27^hi	240.6±80.86^ab	87.74±23.66^defg
	嫩叶	73.21±3.50^g	8.91±1.38^hij	7.49±1.28^cdefg	9.63±3.04^b	13.17±1.94^bcde	1 369.39±328.81ⁱ	135.65±47.36^def	74.71±21.06^ef
	平均值	75.66±3.84	9.33±1.74	7.39±1.10	8.47±2.22	13.35±1.73	1 525.5±315.54	188.13±64.11	81.23±22.36
毛竹 Phyllostachys edulis	功能叶	69.68±3.38^ghi	8.25±0.95^ij	7.28±1.46^defgh	6.64±1.10^defg	14.12±1.55^abcd	1 443.23±423.17ⁱ	101.48±42.52^ef	60.72±9.99^f
	嫩叶	58.37±1.94^k	9.66±1.10^fghi	8.19±1.67^bcdef	4.66±0.92^hijk	13.11±1.88^bcde	1 725.62±456.64^ghi	93.63±23.96^ef	53.92±11.56^f
	平均值	64.03±2.66	8.96±1.10	7.74±1.57	5.65±1.01	13.62±1.72	1 584.43±439.91	97.56±33.24	57.32±10.78
菲黄竹 Pleioblastus auricoma	功能叶	88.39±5.94^cd	10.43±1.25^defgh	8±2.46^bcdefg	5.63±1.24^efghi	15.63±3.18^a	2 814.07±963.67^abcd	203.49±24.49^bc	168.23±24.14^a
	嫩叶	78.31±5.34^f	9.9±1.46^efghi	7.83±0.98^bcdefg	6.06±1.05^defgh	10.71±1.99^fg	1 655.61±426.05^hi	168.23±24.14^cd	153.14±37.89^ab
	平均值	83.35±5.64	10.17±1.36	7.92±1.72	5.85±1.15	13.17±2.59	2 234.84±694.86	185.86±24.32	160.69±31.02
菲白竹 Pleioblastus fortunei	功能叶	69.08±5.99^hi	11.21±2.17^bcdefg	8±1.26^bcdefg	4.92±0.80^ghijk	12.22±1.44^defg	2 554.87±438.61^def	206.77±15.25^bc	113.94±26.19^cde
	嫩叶	66.5±8.40^ij	10.33±1.14^defgh	8.59±0.83^bcde	4.54±0.88^hijk	10.51±1.18^g	2 144.28±590.63^fgh	152.11±32.76^cde	68.55±25.53^ef
	平均值	67.79±7.20	10.77±1.66	8.3±1.05	4.73±0.84	11.37±1.31	2 349.58±514.62	179.44±24.01	91.25±25.86
凤尾竹 Bambusa multiplex f. fernleaf	功能叶	63.51±2.60^j	11.92±1.8^bcd	5.93±0.98^h	3.75±0.82^jk	13.22±1.52^bcde	1 661.61±443.56^hi	94.38±31.72^ef	78.94±79.94^def
	嫩叶	63.47±2.67^j	14.59±2.39^a	7.6±1.42^cdefg	3.46±0.47^k	10.34±1.27^g	1 732.54±192.55^ghi	83.36±33.41^f	71.54±41.67^ef
	平均值	63.49±2.64	13.26±2.10	6.77±1.20	3.61±0.65	11.78±1.40	1 697.08±318.06	88.87±32.57	75.24±60.81
红壳竹 Phyllostachys iridescens	功能叶	86.27±4.87^cde	11.4±1.46^bcdef	7.72±1.93^cdefg	9.00±2.61^bc	14.75±1.10^ab	2 605.30±546.24^cdef	260.29±70.03^ab	119.43±58.38^bcd
	嫩叶	85.94±7.04^cde	9.33±2.33^ghi	8.68±2.03^bcd	6.54±2.59^defg	12.57±1.16^cdef	2 676.27±732.23^bcde	258.08±119.84^ab	103.50±26.59^cdef
	平均值	86.11±5.96	10.37±1.90	8.20±1.98	7.77±2.60	13.66±1.13	2 640.79±639.24	259.19±94.94	111.47±42.49
孝顺竹 Bambusa multiplex	功能叶	89.1±3.75^c	12.47±2.43^bc	9.00±1.30^abc	5.56±1.45^efghi	15.21±2.31^ab	3 056.69±442.96^abc	237.37±69.16^ab	136.23±14.31^abc
	嫩叶	83.96±4.34^e	10.73±1.38^cdefgh	8.39±1.04^bcdef	3.88±1.37^ijk	10.95±1.61^fg	2 208.47±338.79^efg	109.34±47.84^def	62.49±16.68^f
	平均值	86.53±4.05	11.60±1.91	8.70±1.17	4.72±1.41	13.08±1.96	2 632.58±390.88	173.36±58.50	99.36±15.5
金丝慈竹 Bambusa affinis	功能叶	84.22±2.85^gh	10.51±1.34^defgh	6.57±1.36^gh	3.68±0.65^jk	13.83±1.62^abcd	1 717.33±812.86^ghi	291.58±125.08^a	105.40±55.93^cdef
	嫩叶	71.38±3.45^de	13.01±1.73^ab	7.66±1.30^cdefg	3.57±0.82^jk	14.18±3.44^abcd	3 188.70±810.06^a	241.84±96.50^ab	69.19±31.03^ef
	平均值	77.80±3.15	11.76±1.54	7.12±1.33	3.63±0.74	14.01±2.53	2 453.02±811.46	266.71±110.79	87.30±43.48

表1

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竹叶去硅方法的筛选及其细胞学研究

Desilication of bamboo leaves for paraffin slices preparation and their cytological study on eleven bamboo species

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