丹霞地貌铁皮石斛及其人工繁育苗性状比较分析

包英华; 韩伟; 张自斌

doi:10.19303/j.issn.1008-0384.2021.05.007

丹霞地貌铁皮石斛及其人工繁育苗性状比较分析

doi: 10.19303/j.issn.1008-0384.2021.05.007

包英华^1,,
韩伟¹,
张自斌^2, ,

1.
韶关学院英东生物与农业学院，广东　韶关　512005
2.
广西壮族自治区农业科学院花卉研究所，广西　南宁　530007

基金项目: 韶关市科技计划项目（2019sn053）；韶关学院新课程专项（SYJY20181918）；广西重点研发计划项目（桂科AB16380061）

详细信息

作者简介:
包英华（1973−），女，博士，副教授，主要从事珍稀植物种质保育及开发利用研究（E-mail：byhm@163.com）

通讯作者:
张自斌（1978−），男，博士，副教授，主要从事珍稀花卉植物保育和开发应用研究（E-mail：candou154@126.com）

中图分类号: Q 949
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出版历程
- 收稿日期: 2020-07-22
- 修回日期: 2021-03-26
- 网络出版日期: 2021-04-20
- 刊出日期: 2021-05-31

Characteristics of Seedlings of Wild and Cultivated Dendrobium officinale from Areas of Danxia Landform

BAO Yinghua^1
,,
HAN Wei¹,
ZHANG Zibin^{2
, ,}

1.
Henry Fok College of Biology and Agriculture, Shaoguan University, Shaoguan Guangdong　512005, China
2.
Flower Research Institute, Guangxi Academy of Agricultural Sciences, Nanning Guangxi　530007, China

摘要

摘要: 目的以丹霞地貌铁皮石斛和人工繁育苗的2年生和3年生茎作为材料，研究其性状显微特征和多糖含量变化，分析丹霞地貌铁皮石斛在人工繁育和栽培过程中有无品质变化，以期为丹霞地貌铁皮石斛资源的开发利用和人工繁育苗的质量标准制定提供科学依据。方法采用显微性状鉴别、组织化学定位和含量测定方法，对丹霞地貌铁皮石斛与人工繁育苗的性状特征、茎显微构造和多糖的组织化学定位及含量特征进行比较研究。结果丹霞地貌铁皮石斛的茎呈紫红色，叶为紫绿色，节间不明显；人工繁育苗的茎则呈深绿色，叶为浅绿色，节间明显，其株高、茎粗和节间长度均大于丹霞地貌铁皮石斛。两者的角质层厚度、表皮细胞直径、皮下层细胞直径和层数、鞘纤维直径和数目、维管束直径和数目、木质部和韧皮部厚度、草酸钙针晶长度、导管直径、木薄壁细胞直径、鞘细胞壁厚度等均存在显著性差异。丹霞地貌铁皮石斛和人工繁育苗茎的基本组织中分布着大量多糖颗粒，经染色后呈深紫红色，人工繁育苗的多糖颗粒呈现聚集状，被显色更深；多糖颗粒直径与含量间均存在显著差异，丹霞地貌铁皮石斛的总多糖含量高于人工繁育苗。结论丹霞地貌铁皮石斛与人工繁育苗在性状显微特征和代谢产物量上均存在显著差异。其性状特征、显微结构特征、多糖组织化学定位和含量特征均可作为鉴别丹霞地貌铁皮石斛和人工繁育苗的鉴别与评价指标。
- 丹霞地貌铁皮石斛 /
- 人工繁育苗 /
- 性状显微特征 /
- 多糖含量
Abstract: Objective Microstructures and polysaccharide contents in the wild Dendrobium officinalein in Danxia landform areas and artificially propagated seedlings were compared for product differentiation and cultivation of the valuable herbal medicinal material. Methods At areas of Danxia landform, stems from wild and 2-3 year old cultivated D. officinalein plants were collected for microscopic morphological examination as well as histochemical and chemical analyses. Results The reddish-purple stems of the wild D. officinalein plants bore dark greenish-purple leaves with no visible internodes. In contrast, the artificially propagated seedlings had light green leaves on dark green stems with internodes. In addition, the plant height, stem diameter, and internode distance of the cultured seedlings were greater than those of the plants found in the wild. There were also significant differences between them in the cuticle thickness, epidermal cell diameter, subcutaneous cell diameter and layer number, sheath fiber diameter and number, vascular bundle diameter and number, xylem and phloem thickness, calcium oxalate needle length, catheter diameter, wood parenchyma cell diameter, and sheath cell wall thickness of the plants. A large amount of polysaccharide granules existed in aggregation in the tissues of the cultivated seedlings which turned its deep reddish purple color by staining darker than those in the wild D. officinalein. The granules differed in size as well. However, the wild D. officinalein had a higher content of polysaccharides than the propagated seedlings. Conclusion With respect to the microstructure and metabolites of the stems, there were significant differences between the D. officinalein in the wild and the cultivated seedlings. The morphology, microstructure, histochemical localization, and polysaccharide content of the medicinal material could be adequate indicators in identifying, evaluating, and differentiating the products of different origins.
- Dendrobium officinalein Danxia landform /
- artificially propagated seedlings /
- microstructure /
- polysaccharide content

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图 1 丹霞地貌铁皮石斛和人工繁育苗的形态特征

注：1：丹霞地貌铁皮石斛茎；2：丹霞地貌铁皮石斛叶；3：人工繁育苗茎；4：人工繁育苗叶。

Figure 1. Morphology of propagated and wild D. officinale plants from areas of Danxia landform

Note: 1: stem of wild D. officinale; 2: leaf of wild D. officinale; 3: stem of propagated D. officinale seedlings; 4: leaf of propagated D. officinale seedlings.

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图 2 丹霞地貌铁皮石斛和人工繁育苗茎显微结构特征

注：1～3：丹霞地貌铁皮石斛2年生茎；4～6：丹霞地貌铁皮石斛3年生茎；7～9：人工繁育苗2年生茎；10～12：人工繁繁育3年生茎。cu：角质层；ep：表皮；sc：皮下层细胞；gt：基本组织；v：维管束；1、4、7、10：10×4（目镜和物镜）；2、3、5、6、8、9、11、12：10×40（目镜和物镜）。

Figure 2. Microstructures of propagated and wild D. officinale plants from areas of Danxia landform

Note: 1-3: stem of 2-year-old wild D. officinale; 4-6: stem of 3-year-old wild D. officinale; 7-9: stem of 2-year-old propagated D. officinale seedlings; 10-12: stem of 3-year-old propagated D. officinale seedlings. cu: cuticle; ep: epidermal; sc: subcutaneous cell; gt: elementary tissue; v: vascular bundles; 1, 4, 7, and 10: observed under 10×4 (eyepiece × objective); 2, 3, 5, 6, 8, 9, 11, and 12: observed under 10×40 (eyepiece × objective).

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图 3 丹霞地貌铁皮石斛和人工繁育苗茎粉末显微结构特征

注：1：丹霞地貌铁皮石斛2年生茎；2：丹霞地貌铁皮石斛3年生茎；3：人工繁育苗2年生茎；4：人工繁育苗3年生茎。A：草酸钙针晶；B：导管；C：薄壁细胞；D：鞘细胞；E：淀粉粒。

Figure 3. Microstructures of powdered stem of propagated and wild D. officinale plants from areas of Danxia landform

Note: 1: stem of 2-year-old wild D. officinale; 2: stem of 3-year-old wild D. officinale; 3: stem of 2-year-old propagated D. officinale seedlings; 4: stem of 3-year-old propagated D. officinale seedlings. A: calcium oxalate crystal; B: vessel; C: parenchyma cell; D: sheath cell; E: starch granule.

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图 4 丹霞地貌铁皮石斛和人工繁育苗茎多糖组织化学定位

注：1～3：丹霞地貌铁皮石斛2年生茎；4～6：丹霞地貌铁皮石斛3年生茎；7～9：人工繁育苗2年生茎；10～12：人工繁育苗3年生茎。p：多糖颗粒；1、4、7、10：10×4（目镜和物镜）；2、3、5、6、8、9、11、12：10×40（目镜和物镜）。

Figure 4. Histochemical localization of polysaccharides in stems of propagated and wild D. officinale plants from areas of Danxia landform

Note: 1-3: stem of 2-year-old wild D. officinale; 4-6: stem of 2-year-old wild D. officinale; 7-9: stem of 2-year-old propagated D. officinale seedlings; 10-12: stem of 3-year-old propagated D. officinale seedlings. P: polysaccharide; 1, 4, 7, and 10: observed under 10×4 (eyepiece × objective); 2, 3, 5, 6, 8, 9, 11, and 12: observed under 10×40 (eyepiece × objective).

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表 1 丹霞地貌铁皮石斛和人工繁育苗的形态特征分析

Table 1. Morphology of propagated and wild D. officinale plants from areas of Danxia landform

材料 Material	株高 Plant height/cm	茎直径 Stemdiameter/ mm	节间长度 Internode length/cm	叶片长度 Leaf blade length/cm	叶片宽度 Leaf blade width/cm
2年生丹霞地貌铁皮石斛苗 2 years old（Dendrobium officinale in Danxia landform）	13.91±2.21 b	3.74±0.76 b	0.22±0.11 b	3.85±0.58	1.10±0.21
3年生丹霞地貌铁皮石斛苗 3 years old（Dendrobium officinale in Danxia landform）	12.15±1.70 b	3.30±0.49 b	0.18±0.09 b	—	—
2年生人工繁育苗 2 years old（artificially bred）	12.15±1.70 b	9.79±1.21 a	0.62±0.24 a	6.77±1.54	2.14±0.36
3年生人工繁育苗 3 years old（artificially bred）	18.71±1.67 a	9.42±1.10 a	0.55±0.18 a	—	—
注：同列数据后不同小写字母表示各处理之间的差异显著性（P＜0.05）。下同。 Note: Data with different lowercase letters on the same column indicate significance of the difference at P＜0.05.The same as below.

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表 2 丹霞地貌铁皮石斛和人工繁育苗的茎显微结构特征比较分析

Table 2. Microstructures of stems of propagated and wild D. officinale plants from areas of Danxia landform

材料 Material	角质层厚度 Cuticle thickness/ µm	表皮细胞直径 Epidermal cell diameter/ µm	皮下层细胞直径 Subcutaneous cell diameter/ µm	皮下层细胞层数 The number of layers of subcutaneous cell	鞘纤维直径 Sheath fiber diameter/ µm	维管束直径µm Vascular bundle diameter/ µm	鞘纤维数目 Number of sheath fibers	维管束数目 Numberof vascular bundles	木质部厚度 Xylem thickness/ µm	韧皮部厚度 Phloem thickness/ µm
2年生丹霞地貌铁皮石斛苗 2 years old（Dendrobium officinale in Danxia landform）	8.58±1.98 c	17.85±4.84 c	28.56±8.16 c	1.67±0.51 b	28.99±11.07 ab	13.55±2.28 b	151.15±0.62 c	54.4±9.46 a	31.21±11.79 b	34.81±8.34 b
3年生丹霞地貌铁皮石斛苗 3 years old（Dendrobium officinale in Danxia landform）	10.85±4.18 a	22.23±4.48 a	28.56±8.16 c	1.55±0.65 b	30.66±8.63 a	15.22±3.02 a	171.53±40.5 a	42.75±11.14 b	38.88±15.35 a	44.12±22.47 a
2年生人工繁育苗 2 years old（artificially bred）	9.11±2.07 bc	20.85±4.41 ab	35.15±8.71 b	1.57±0.62 b	26.44±7.92 b	14.67±3.06 b	141.36±37.94 ab	39.1±8.24 c	32.21±2.1 b	35.33±12.44 b
3年生人工繁育苗 3 years old（artificially bred）	9.86±1.58 b	20.16±4.18 b	39.84±10.53 a	2.02±0.34 a	28.34±5.03 ab	14.02±2.09 b	151.06±23.39 bc	34.35±10.7 d	32.82±5.74 ab	40.02±8.77 b

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表 3 丹霞地貌铁皮石斛和人工繁育苗茎粉末显微结构特征比较分析

Table 3. Microstructures of powdered stems of propagated and wild D. officinale plants from areas of Danxia landform

材料 Material	草酸钙针晶长度 Calcium oxalate c rystal length/ µm	草酸钙针晶直直径 Calcium oxalate crystaldiameter/ µm	导管直径 Vessel diameter/ µm	木薄壁细胞直径 Wood parenchyma cell diameter /µm	鞘细胞壁厚度 Thickness of sheath cell wall/ µm	淀粉粒直径 Starch grain diameter/ µm
2年生丹霞地貌铁皮石斛苗 2 years old （Dendrobium officinale in Danxia landform）	52.25±24.06 c	2.07±0.57 a	9.64±4.63 b	59.08±29.64 a	5.34±1.07 b	7.95±1.28 a
3年生丹霞地貌铁皮石斛苗 Th3 years oldr（Dendrobium officinale in Danxia landform）form）	58.81±5.09 ab	2.20±0.44 a	8.55±2.82 b	49.79±17.13 ab	5.45±1.14 b	7.28±2.10 a
2年生人工繁育苗 2 years old（artificially bred）	70.13±7.87 a	1.93±0.59 a	11.81±4.71 ab	34.48±5.98 bc	5.97±1.41 ab	7.90±2.34 a
3年生人工繁育苗 3 years old（artificially bred）	68.79±12.44 ab	1.79±0.63 a	13.85±5.91 a	34.01±17.76 c	6.39±1.20 a	7.93±1.70 a

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表 4 丹霞地貌铁皮石斛和人工繁育苗茎多糖的组织化学定位与含量分析

Table 4. Histochemical localization and polysaccharide content in stems of propagated and wild D. officinale plants from areas of Danxia landform

材料 Material	多糖颗粒直径 Polysaccharide diameter/µm	总多糖含量 Total polysaccharide content/%
2年生丹霞地貌铁皮石斛苗 2 years old（Dendrobium officinale in Danxia landform）	10.98±6.97 c	35.46±2.11 a
3年生丹霞地貌铁皮石斛苗 3 years old（Dendrobium officinale in Danxia landform）	12.90±6.88 c	32.37±2.02 b
2年生人工繁育苗 2 years old（artificially bred）	18.73±21.96 b	12.24±0.69 d
3年生人工繁育苗 3 years old（artificially bred）	28.61±15.32 a	18.02±0.60 c

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参考文献(15)

[1]	中华人民共和国药典委员会. 中国药典: 一部[M]. 北京: 化学工业出版社, 2015: 282.
[2]	中国科学院植物研究所. 中国高等植物图鉴: 第5册[M]. 北京: 科学出版社, 1980.
[3]	中国科学院中国植物志编辑委员会. 中国植物志: 第19卷[M]. 北京: 科学出版社, 1999.
[4]	包英华, 韩伟, 白音, 等. ‘丹霞’铁皮石斛的组织培养与快速繁殖 [J]. 分子植物育种, 2019, 17(5):1616−1623. BAO Y H, HAN W, BAI Y, et al. Tissue culture and rapid propagation of ‘Danxia’ Dendrobium officinale [J]. Molecular Plant Breeding, 2019, 17(5): 1616−1623.(in Chinese)
[5]	黄民权, 卢应京. 石斛愈伤组织培养物的药用前景探讨 [J]. 中药材, 1998(11):543−545. doi: 10.3321/j.issn:1001-4454.1998.11.002 HUANG M Q, LU Y J. Prospect on the callus culture of dentrobium candidum used as drugs [J]. Journal of Chinese Medicinal Materials, 1998(11): 543−545.(in Chinese) doi: 10.3321/j.issn:1001-4454.1998.11.002
[6]	范俊安, 王继生, 张艳, 等. 铁皮石斛组培品与野生品的形态组织学和多糖含量比较研究 [J]. 中国中药杂志, 2005(21):1648−1650, 1659. doi: 10.3321/j.issn:1001-5302.2005.21.002 FAN J A, WANG J S, ZHANG Y, et al. A comparison of tissue formation and the content of polysaccharide between wild and cultured Dendrobium candidum [J]. China Journal of Chinese Materia Medica, 2005(21): 1648−1650, 1659.(in Chinese) doi: 10.3321/j.issn:1001-5302.2005.21.002
[7]	黎万奎, 胡之璧, 周吉燕, 等. 人工栽培铁皮石斛与其他来源铁皮石斛中氨基酸与多糖及微量元素的比较分析 [J]. 上海中医药大学学报, 2008(4):80−83. doi: 10.3969/j.issn.1008-861X.2008.04.021 LI W K, HU Z B, ZHOU J Y, et al. Comparative analysis of amino acids, polysaccharides and trace elements in Dendrobium officinale from different sources [J]. Acta Universitatis Traditionis Medicalis Sinensis Pharmacologiaeque Shanghai, 2008(4): 80−83.(in Chinese) doi: 10.3969/j.issn.1008-861X.2008.04.021
[8]	付涛, 王志龙. GC-MS法分析不同部位仿野生与组培快繁铁皮石斛中的挥发性成分 [J]. 中成药, 2015, 37(12):2702−2709. FU T, WANG Z L. Analysis of volatile constituents in different parts of imitative wild and tissue culture and rapid propagation of Dendrobium officinale by GC-MS [J]. Chinese Traditional Patent Medicine, 2015, 37(12): 2702−2709.(in Chinese)
[9]	包英华, 白音, 石海英. 金钗石斛多糖的组织化学定位研究 [J]. 时珍国医国药, 2012, 23(12):3026−3028. doi: 10.3969/j.issn.1008-0805.2012.12.035 BAO Y H, BAI Y, SHI H Y. Histochemistry localization for polysaccharide in Dendrobium nobile lindl [J]. Lishizhen Medicine and Materia Medica Research, 2012, 23(12): 3026−3028.(in Chinese) doi: 10.3969/j.issn.1008-0805.2012.12.035
[10]	赵菊润, 张治国. 铁皮石斛产业发展现状与对策 [J]. 中国现代中药, 2014, 16(4):277−279, 286. ZHAO J R, ZHANG Z G. Development status, issues and countermeasures of Dendrobium officinale industry [J]. Modern Chinese Medicine, 2014, 16(4): 277−279, 286.(in Chinese)
[11]	周玉飞, 康专苗, 彭竹晶. 珍稀濒危铁皮石斛的研究进展 [J]. 基因组学与应用生物学, 2018, 37(4):1629−1635. ZHOU Y F, KANG Z M, PENG Z J. Research progress of rare and endangered plant Dendrobium officinale [J]. Genomics and Applied Biology, 2018, 37(4): 1629−1635.(in Chinese)
[12]	秦文, 李旭梅, 卫子皎, 等. 铁皮石斛的性状和显微鉴别研究 [J]. 时珍国医国药, 2017, 28(8):1913−1915. QIN W, LI X M, WEI Z J, et al. Study on characters and microscopic identification of Dendrobium officinale [J]. Lishizhen Medicine and Materia Medica Research, 2017, 28(8): 1913−1915.(in Chinese)
[13]	刘富裕, 郭庆梅. 激光显微切割技术在药用植物学研究中的应用 [J]. 植物生理学报, 2019, 55(11):1563−1569. LIU F Y, GUO Q M. Research progress of laser microdissection technique for medicinal botany [J]. Plant Physiology Journal, 2019, 55(11): 1563−1569.(in Chinese)
[14]	林丽, 施晓龙, 高素芳, 等. 显微组织化学定位在中药研究中的应用 [J]. 中国现代中药, 2014, 16(6):505−509. LIN L, SHI X L, GAO S F, et al. The application of microscopic histochemical localization in traditional Chinese medicine study [J]. Modern Chinese Medicine, 2014, 16(6): 505−509.(in Chinese)
[15]	龚庆芳, 周浩, 王新桂, 等. 不同产地铁皮石斛的品质比较研究 [J]. 北方园艺, 2013(8):162−165. GONG Q F, ZHOU H, WANG X G, et al. Comparative study on quality of Dendrobium officinale from different habits [J]. Northern Horticulture, 2013(8): 162−165.(in Chinese)