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

doi:10.3969/j.issn.1000-2006.201903003

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2019, Vol. 43 ›› Issue (5): 74-80.doi: 10.3969/j.issn.1000-2006.201903003

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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
Contact: LIN Shuyan E-mail:1036654249@qq.com;lrx@njfu.com.cn

Abstract

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

CLC Number:

S795

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, 2019, 43(5): 74-80.

Figures/Tables 4

Fig.1

Fig.2

Fig.3

Table 1

Comparison of morphological and anatomical features of leaves of 11 bamboo species"

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

Table 1

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

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