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Variety	Phenotypic characteristic	Average yield	Quality characteristic	Variety advantage	Source	Breeding method
Tianhu 1^[43]	SH 20-60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots	0.91 kg·m^-2 (20 months growth, fresh weight)	DR 15.5%, PC 15.6%	Good comprehensive characters, higher quality, better low- temperature and disease resistant	Domestication of wild resources	Selective breeding
Xianhu 1^[45]	SH 20-30 cm, up to 60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots	2.29 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 24.6%, PC 30.4%	Higher yield and PC, better adverse resistance	Domestication of wild resources Sxg-2	Selective breeding
Xianhu 2^{[46, 47]}	SH 30-50 cm, SD 6-10 mm, ID 15-25 mm, stem with purple spots; LL 4-7 cm, LW 1-2.5 cm, LT 0.6-0.9 mm, LS with purple spots	2.49 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 21.1%, PC 29.0%	Higher yield and PC, better low- temperature and disease resistant	Domestication of wild resources Xianzi 1	Selective breeding
Xianhu 3^[48]	SH 8-21 cm, SD 2-6 mm, ID 8-14 mm; LL 2-7 cm, LW 0.8-2.2 cm, edge and main vein of leaves often with purple spots	2.52 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 33.4%, EEC 13.85%, MC 21.75%, R_M/G 4.42	Higher yield, better adaptability and adverse resistance	MP: Xianhu1; FP: No. 514	Cross breeding and mutation breeding
Senshan 1^[49]	SH 14-36.6 cm, SD 3.58-6 mm; LL 3.8-5.3 cm, LW 1.7-2.2 cm, leaf back and LS often with purple spot	1.35 kg·m^-2 (2 years growth)	PC 24.45%	Higher yield, better adaptability	Domestication of wild resources collected from Yunnan	Selective breeding
Guihu 1^{[50, 51]}	SH 15-60 cm, SD 4-7.2 mm, stem with purple red spots/patches; LL 2-7 cm, LW 0.5-2.5 cm, leaf and LS with purple red spots/patches	0.23-0.64 kg·m^-2 (18-20 months growth, the first harvest); 0.38-0.76 kg·m^-2 (harvest once every 12 to 14 months after the first harvest, the second and third one)	PC 28.0%- 35.9% (fresh stems)	Better drought and root rot resistance	Selection of good individual from wild seedlings (Nazuo Miao Nationality Township, Xilin County, Guangxi)	Selective breeding
Baishishan 1^[52]	SH 12-65 cm, SD 4-8 mm, ID 1.5-2.5 cm, stem with purple spots; LL 4-6.5 cm, LW 1.3-2.5 cm, LT 7-9 mm, leaf edge often pale purple, leaf back often with pale purple spots	0.30-0.68 kg·m^-2 (18-24 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)	PC > 29% (dry stem)	/	Selection of good individual from test tube seedlings (purple stem of wild adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)	Selective breeding
Baishishan 2^[53]	SH 15-70 cm, SD 4-9 mm, ID 2-3 cm, no spots on stem; LL 4-6.5 cm, LW 1.2-2.5 cm, LT 7-9 mm	0.30-0.76 kg·m^-2 (18-20 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)	PC about 31%	/	Selection of good individual from wild seedlings (green stem of adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)	Selective breeding
Qinggu 1^[54]	SH 35-37 cm, SD 10.5-11.5 mm, stem full with purple spots; leaf edge purple, leaf back with purple spots	0.69 kg·m^-2 (fresh stems, 2 years growth) 0.72 kg·m^-2 (fresh stems, 3 years growth)	PC 47.7% (2 years growth); PC 47.2% (3 years growth)	/	Collection of wild resources from Yunnan; then selection good single-plant	Selective breeding
Zhongke 1^{[55, 56]}	SH up to 50 cm, SD up to 7 mm; leaves rectangular and lanceolate, LL 5-6 cm, LW 2-3 cm, LS with rust spots	1.5 kg·m^-2 (18 months growth of tissue cultured seedling)	PC about 30% (dry stems)	Higher yield, better high-temperature and disease resistance	T13 (Guangnan County, Yunnan), after multiple generations of self-breeding	Selective breeding
Shuanghui 1^[57-59]	SH 30-50 cm, SD 5-8 mm, ID 1.5-2.5 cm; leaves spindle, LL 6.0 cm, LW 2.0 cm	1.4 kg·m^-2 (fresh stem, 2 years growth)	PC 32.7%, MC 19.2%	Higher yield, better disease and adverse resistance	FP: red stem (Yandang Mountains, Zhejiang), MP: green stem (Guangnan County, Yunnan); artificial cross-pollination, single- plant selection of offspring, multiple generations of selfing purification	Cross breeding
Zhongke Congdu^{[60, 61]}	SH up to 55 cm and SD 6 mm after 2 years growth; leaves rectangular and lanceolate, LL 4-6 cm, LW 2-3 cm, LS with rust spots	1.2 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 32.1%, MC 23.7% (dry stem)	Higher PC, better adverse resistance	Introduction of the plants from Yachang orchid plant nature reserve (Leye County, Guangxi); then selection from multiple generations of self-breeding	Selective breeding
Zhongke Congdu 2^[62]	SH 34.3 cm and SD 6 mm after 2 years growth	1.36 kg·m^-2 (fresh stems, 18 months growth of tissue cultured seedling)	PC 42.4%, MC 29.4%, EEC 13.0%, R_M/G 2.6 (dry stem)	Higher PC, better disease resistance	Wild resources collected from Dao County, Hunan; selection after 4 generations of self-breeding	Selective breeding
Yanchuixue 3^{[62, 63]}	SH 25.5 cm and SD 6 mm after 2 years growth, stem with rust spots; LL 4.8 cm, LW 1.5 cm	0.62 kg·m^-2 (fresh stems)	PC 57.5%, MC32.6%, R_M/G 2.3 (dry stem)	Better disease resistance	Hybrid of green stem (Tiantai Mountains) and red stem (Yandang Mountains) plants	Cross breeding
Zhongke 3^[64]	SH 36.2 cm and SD 4.7 mm after 2 years growth, stem with rust spots; leaves narrow ovate, LL 5.1 cm, LW 1.6 cm	1.45 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 39.4%, MC 18.6%, EEC 6.6% (dry stem)	Better disease and adverse resistance	Wild resource T636 (Dao County, Hunan); selection after 4 generations of self-breeding	Selective breeding
Zhongke 4^[64]	SH 48.0 cm and SD 5 mm after 2 years growth, stem with rust spots; leaves ovate, LL 5.7 cm, LW 1.9 cm	1.56 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 39.0%, MC 20.1%, EEC 6.6% (dry stem)	Better disease and adverse resistance	Wild resource T709 (Lang Mountains, Xinning County, Hunan); selection after 4 generations of self-breeding	Selective breeding
Fuhu 1^{[65, 66]}	SH 20.8 cm, SD 5.1 mm and ID 1.5 cm after 2 years growth, stem with pale purple spots; leaves long elliptic, LL 5 cm, LW 1.6 cm	0.48 kg·m^-2 (fresh stem)	PC 51.8%, MC 29.3%, R_M/G 2.5 (dry stem)	Higher yield and quality, lower incidence of black spot and anthracnose	Selection of wild resource L-2 from Guaizhai Mountains, Liancheng County, Fujian	Selective breeding

Variety	Phenotypic characteristic	Average yield	Quality characteristic	Variety advantage	Source	Breeding method
Tianhu 1^[43]	SH 20-60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots	0.91 kg·m^-2 (20 months growth, fresh weight)	DR 15.5%, PC 15.6%	Good comprehensive characters, higher quality, better low- temperature and disease resistant	Domestication of wild resources	Selective breeding
Xianhu 1^[45]	SH 20-30 cm, up to 60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots	2.29 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 24.6%, PC 30.4%	Higher yield and PC, better adverse resistance	Domestication of wild resources Sxg-2	Selective breeding
Xianhu 2^{[46, 47]}	SH 30-50 cm, SD 6-10 mm, ID 15-25 mm, stem with purple spots; LL 4-7 cm, LW 1-2.5 cm, LT 0.6-0.9 mm, LS with purple spots	2.49 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 21.1%, PC 29.0%	Higher yield and PC, better low- temperature and disease resistant	Domestication of wild resources Xianzi 1	Selective breeding
Xianhu 3^[48]	SH 8-21 cm, SD 2-6 mm, ID 8-14 mm; LL 2-7 cm, LW 0.8-2.2 cm, edge and main vein of leaves often with purple spots	2.52 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 33.4%, EEC 13.85%, MC 21.75%, R_M/G 4.42	Higher yield, better adaptability and adverse resistance	MP: Xianhu1; FP: No. 514	Cross breeding and mutation breeding
Senshan 1^[49]	SH 14-36.6 cm, SD 3.58-6 mm; LL 3.8-5.3 cm, LW 1.7-2.2 cm, leaf back and LS often with purple spot	1.35 kg·m^-2 (2 years growth)	PC 24.45%	Higher yield, better adaptability	Domestication of wild resources collected from Yunnan	Selective breeding
Guihu 1^{[50, 51]}	SH 15-60 cm, SD 4-7.2 mm, stem with purple red spots/patches; LL 2-7 cm, LW 0.5-2.5 cm, leaf and LS with purple red spots/patches	0.23-0.64 kg·m^-2 (18-20 months growth, the first harvest); 0.38-0.76 kg·m^-2 (harvest once every 12 to 14 months after the first harvest, the second and third one)	PC 28.0%- 35.9% (fresh stems)	Better drought and root rot resistance	Selection of good individual from wild seedlings (Nazuo Miao Nationality Township, Xilin County, Guangxi)	Selective breeding
Baishishan 1^[52]	SH 12-65 cm, SD 4-8 mm, ID 1.5-2.5 cm, stem with purple spots; LL 4-6.5 cm, LW 1.3-2.5 cm, LT 7-9 mm, leaf edge often pale purple, leaf back often with pale purple spots	0.30-0.68 kg·m^-2 (18-24 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)	PC > 29% (dry stem)	/	Selection of good individual from test tube seedlings (purple stem of wild adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)	Selective breeding
Baishishan 2^[53]	SH 15-70 cm, SD 4-9 mm, ID 2-3 cm, no spots on stem; LL 4-6.5 cm, LW 1.2-2.5 cm, LT 7-9 mm	0.30-0.76 kg·m^-2 (18-20 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)	PC about 31%	/	Selection of good individual from wild seedlings (green stem of adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)	Selective breeding
Qinggu 1^[54]	SH 35-37 cm, SD 10.5-11.5 mm, stem full with purple spots; leaf edge purple, leaf back with purple spots	0.69 kg·m^-2 (fresh stems, 2 years growth) 0.72 kg·m^-2 (fresh stems, 3 years growth)	PC 47.7% (2 years growth); PC 47.2% (3 years growth)	/	Collection of wild resources from Yunnan; then selection good single-plant	Selective breeding
Zhongke 1^{[55, 56]}	SH up to 50 cm, SD up to 7 mm; leaves rectangular and lanceolate, LL 5-6 cm, LW 2-3 cm, LS with rust spots	1.5 kg·m^-2 (18 months growth of tissue cultured seedling)	PC about 30% (dry stems)	Higher yield, better high-temperature and disease resistance	T13 (Guangnan County, Yunnan), after multiple generations of self-breeding	Selective breeding
Shuanghui 1^[57-59]	SH 30-50 cm, SD 5-8 mm, ID 1.5-2.5 cm; leaves spindle, LL 6.0 cm, LW 2.0 cm	1.4 kg·m^-2 (fresh stem, 2 years growth)	PC 32.7%, MC 19.2%	Higher yield, better disease and adverse resistance	FP: red stem (Yandang Mountains, Zhejiang), MP: green stem (Guangnan County, Yunnan); artificial cross-pollination, single- plant selection of offspring, multiple generations of selfing purification	Cross breeding
Zhongke Congdu^{[60, 61]}	SH up to 55 cm and SD 6 mm after 2 years growth; leaves rectangular and lanceolate, LL 4-6 cm, LW 2-3 cm, LS with rust spots	1.2 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 32.1%, MC 23.7% (dry stem)	Higher PC, better adverse resistance	Introduction of the plants from Yachang orchid plant nature reserve (Leye County, Guangxi); then selection from multiple generations of self-breeding	Selective breeding
Zhongke Congdu 2^[62]	SH 34.3 cm and SD 6 mm after 2 years growth	1.36 kg·m^-2 (fresh stems, 18 months growth of tissue cultured seedling)	PC 42.4%, MC 29.4%, EEC 13.0%, R_M/G 2.6 (dry stem)	Higher PC, better disease resistance	Wild resources collected from Dao County, Hunan; selection after 4 generations of self-breeding	Selective breeding
Yanchuixue 3^{[62, 63]}	SH 25.5 cm and SD 6 mm after 2 years growth, stem with rust spots; LL 4.8 cm, LW 1.5 cm	0.62 kg·m^-2 (fresh stems)	PC 57.5%, MC32.6%, R_M/G 2.3 (dry stem)	Better disease resistance	Hybrid of green stem (Tiantai Mountains) and red stem (Yandang Mountains) plants	Cross breeding
Zhongke 3^[64]	SH 36.2 cm and SD 4.7 mm after 2 years growth, stem with rust spots; leaves narrow ovate, LL 5.1 cm, LW 1.6 cm	1.45 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 39.4%, MC 18.6%, EEC 6.6% (dry stem)	Better disease and adverse resistance	Wild resource T636 (Dao County, Hunan); selection after 4 generations of self-breeding	Selective breeding
Zhongke 4^[64]	SH 48.0 cm and SD 5 mm after 2 years growth, stem with rust spots; leaves ovate, LL 5.7 cm, LW 1.9 cm	1.56 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 39.0%, MC 20.1%, EEC 6.6% (dry stem)	Better disease and adverse resistance	Wild resource T709 (Lang Mountains, Xinning County, Hunan); selection after 4 generations of self-breeding	Selective breeding
Fuhu 1^{[65, 66]}	SH 20.8 cm, SD 5.1 mm and ID 1.5 cm after 2 years growth, stem with pale purple spots; leaves long elliptic, LL 5 cm, LW 1.6 cm	0.48 kg·m^-2 (fresh stem)	PC 51.8%, MC 29.3%, R_M/G 2.5 (dry stem)	Higher yield and quality, lower incidence of black spot and anthracnose	Selection of wild resource L-2 from Guaizhai Mountains, Liancheng County, Fujian	Selective breeding

铁皮石斛种质资源和遗传育种研究进展

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陈晓梅 ^* , 田丽霞 , 单婷婷 , 孙乐 , 郭顺星 ^*

药学学报 | 综述 2018,53(9): 1493-1503

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药学学报 | 综述 2018, 53(9): 1493-1503

铁皮石斛种质资源和遗传育种研究进展

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陈晓梅^*, 田丽霞, 单婷婷, 孙乐, 郭顺星^*

作者信息

中国医学科学院、北京协和医学院药用植物研究所, 北京 100193

通讯作者:

* 陈晓梅, Tel:86-10-57833241, E-mail:cxm_implad@163.com;

郭顺星, Tel:86-10-57833231, E-mail:sxguo1986@163.com

Advances in germplasm resources and genetics and breeding of Dendrobium officinale

Xiao-mei CHEN^*, Li-xia TIAN, Ting-ting SHAN, Le SUN, Shun-xing GUO^*

Affiliations

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China

出版时间: 2018-09-12 doi: 10.16438/j.0513-4870.2018-0286

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摘要

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铁皮石斛是一种兰科石斛属药用植物，其干燥茎为名贵中药铁皮石斛，具有益胃生津、滋阴清热的功效。铁皮石斛自然资源濒临枯竭，依赖人工种植生产药材，目前种植面积已经超过10万亩，年产量万吨以上。优良品种是保障中药品质的前提，种质资源是选育优良品种的基础。本文归纳了现有主要铁皮石斛品种的特点，对铁皮石斛种质资源和遗传育种的研究进展进行了综述，期望能够为这两个方面的深入研究提供科学依据。

关键词

铁皮石斛 / 种质资源 / 遗传育种 / 品种

Abstract

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Dendrobium officinale is a member of the family Orchidaceae. The dried stem of D. officinale is used as a valuable traditional Chinese medicine, known as Dendrobii Officamlis Caulis (called TiepiShihu in Chinese). According to Chinese Pharmacopoeia, Dendrobii Officamlis Caulis has effects of tonifying stomach, promoting fluid, nourishing Yin and clearing heat. At present, the planting area of D. officinale is over 100 000 Mu (over 6 670 hm²) and the annual output of its fresh stem is in excess of 10 000 tons. Good variety is the guarantee of herbal medicine's quality, while germplasm resource is the base for breeding excellent variety. In this paper, we summed the characteristics of present main varieties of D. officinale and reviewed the progress on germplasm resources and genetics and breeding of the plant, in order to provide a scientific basis for the further research.

Key words

Dendrobium officinale / germplasm resources / genetics and breeding / variety

引用本文

陈晓梅, 田丽霞, 单婷婷, 孙乐, 郭顺星. 铁皮石斛种质资源和遗传育种研究进展. 药学学报, 2018 , 53 (9) : 1493 -1503 . DOI: 10.16438/j.0513-4870.2018-0286

Xiao-mei CHEN, Li-xia TIAN, Ting-ting SHAN, Le SUN, Shun-xing GUO. Advances in germplasm resources and genetics and breeding of Dendrobium officinale[J]. Acta Pharmaceutica Sinica, 2018 , 53 (9) : 1493 -1503 . DOI: 10.16438/j.0513-4870.2018-0286

正文

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铁皮石斛(Dendrobium officinale Kimura et Migo)是一种兰科石斛属的濒危药用植物, 其干燥茎为中药铁皮石斛, 具有益胃生津、滋阴清热的功效。药理学研究表明铁皮石斛具有抗肿瘤、增强机体免疫力、有效减轻肝损伤等作用^[1]。由于受到自然和人为因素的影响, 铁皮石斛野生资源遭到严重破坏, 已濒临灭绝。随着铁皮石斛人工繁育技术的不断成熟和发展, 从上世纪90年代至今, 铁皮石斛在我国的种植范围迅速扩大, 种植面积已经超过10万亩(1亩= 660 m²), 年产量万吨以上。在种植业的带动下, 铁皮石斛产业得以逐渐形成并快速发展, 目前的产业规模已达到百亿元^[2]。

铁皮石斛种植业存在盲目引种、种质混杂的问题, 导致铁皮石斛质量、产量差异大, 已经影响了产业的正常、有序发展。Tian等^[1]报道除农艺性状的差异外, 红杆和绿杆两种表型铁皮石斛的醇溶性浸出物、多糖和联苄化合物含量, 以及它们对菌根真菌的响应能力都有明显差异。与种植的快速发展相比, 铁皮石斛种质资源和品种选育相关的研究起步晚, 发展较缓慢。一方面, 铁皮石斛野生资源储量的急剧下降及分布范围的迅速缩小, 对系统开展铁皮石斛种质资源的收集、保存和评价等工作造成严重影响; 另一方面, 铁皮石斛遗传基础的研究薄弱, 缺乏从育种角度评价和利用野生资源的基础性研究, 导致品种选育和改良的工作缺少理论支持, 育种目标不明确^{[3, 4]}。本文归纳了现有主要铁皮石斛品种的特征特性, 对铁皮石斛种质资源和遗传育种的研究进展进行了综述, 期望为这两方面的深入研究提供参考, 进而为铁皮石斛种植业以及铁皮石斛产业的持续、快速发展提供有力支撑。

1 铁皮石斛种质资源研究

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种质资源是发现、选择和培育优良品种的基础。研究药用植物种质资源具有特殊意义, 它是保障中药材质量, 保证中药临床疗效和用药安全的前提。近年来, 受气候条件变化、生存环境恶化和人为过度采挖等因素的影响, 野生铁皮石斛种质资源急剧减少, 这将对铁皮石斛的优良品种选育、规范化生产和临床安全使用造成影响。铁皮石斛种质资源的研究归纳起来主要有三个方面:表型性状、种质亲缘关系和种质鉴别技术。表型性状的研究能为人工栽培铁皮石斛的选种和育种提供一定参考。亲缘关系和种质鉴别技术的研究可以为铁皮石斛核心种质资源的筛选和保护, 以及优良品种的选育、保护和推广使用奠定科学基础, 提供技术保障。这两方面的研究主要是依靠分子生物学和生物信息学的技术方法来完成的, 包括了基因型稳定性和多态性检测、遗传多样性评价和物种的种内及种间关系研究等内容^[5]。

1.1 铁皮石斛表型性状研究

铁皮石斛表型性状多样性较高, 通常考察的指标包括:茎长、茎粗、叶长、叶宽、叶面积等。叶面积是影响植物光合速率的主要因素, 是选择优良家系时的重要参考指标之一。Zhu等^[6]研究发现铁皮石斛单片叶面积与叶长宽积之间存在良好的线性关系, 判定系数0.987 7, 茎叶片数与茎总叶面积之间存在幂函数关系, 判定系数0.793 1;建立叶面积回归模型可以在铁皮石斛生长过程中快速估算杂交F₁代家系的叶面积, 为品种选育提供依据。将其用于铁皮石斛种质遗传改良的早代测验, 可以提高后期选择的针对性, 节省时间和人力。铁皮石斛以茎入药, 单位面积的茎数量与单株茎重量是决定产量的重要因素。Zhang等^[7]报道, 铁皮石斛茎长的变异系数最高, 达到44.0%, 茎节数和叶面积的变异系数30%~35%, 节距、茎粗、叶长和叶形指数的变异系数20%~30%, 叶宽的变异系数最小, 为18.8%;产量相关的两个性状指标, 茎长和茎粗, 都与叶片宽度呈显著正相关, 茎长还与茎节数、节距、叶片长度、叶面积和叶形指数呈显著正相关, 茎粗与茎长和节距呈显著负相关; 茎较长的种质叶形细长, 节距较长, 茎较细; 茎较粗的种质叶片较宽, 节距较短。

栽培环境对铁皮石斛的表型性状和有效成分含量会产生较大影响。在野外环境中生长有利于铁皮石斛形成并积累次生代谢产物。相同品系的铁皮石斛, 分别在设施盆栽、岩壁附生和梨树附生的条件下生长1年后取样观察, 与设施盆栽铁皮石斛相比, 岩壁附生和梨树附生铁皮石斛植株粗壮、矮小, 地上部分逐渐变成紫红色, 生物量明显降低; 叶片稀少, 叶伸展角度减小, 叶片厚度、叶肉厚度和上表皮厚度明显增加, 气孔密度明显增大, 气孔周长减小; 根系更发达, 根被细胞层数减少, 细胞壁明显加厚, 根被层内细胞较小, 但排列紧密; 岩壁附生铁皮石斛醇溶性浸出物含量和多糖含量均最高, HPLC指纹图谱分析醇溶性物质的化学成分最复杂, 其次为梨树附生铁皮石斛^[8]。

“口嚼无渣, 黏性强”是优质铁皮石斛的传统判定标准, 据此可以将人工栽培铁皮石斛分为“软脚”和“硬脚”两类。“软脚” (或“F型”、“枫斗型”)铁皮石斛茎较短, 柔软, 具黏性, 基本组织富含多糖, 无下皮层或下皮层不发达, 横切面上维管束数目较少, 维管束鞘纤维不发达, 纤维素含量较低, 适宜加工“枫斗”; “硬脚” (或“H型”、非枫斗型)铁皮石斛茎较长, 黏性差, 具厚壁细胞组成的发达下皮层, 横切面上维管束数目较多, 维管束鞘纤维发达, 纤维素含量较高, 不适宜加工“枫斗”^{[9, 10]}。通过对F型与H型居群铁皮石斛rDNA ITS区碱基序列的测定和分析, 在ITS1区及5.8S区各发现1个序列差异位点^[11], 证明铁皮石斛的表型差异具有遗传基础。

《药典》规定的与铁皮石斛质量相关的考察指标主要有醇溶性浸出物含量, 以及糖类相关的多糖含量、甘露糖含量、甘露糖与葡萄糖色谱峰面积比等。Luo等^[12]分析不同种源铁皮石斛农艺性状、糖类成分含量、叶绿素含量发现:叶长与茎粗呈显著正相关; 茎粗和叶长两项性状指标与多糖含量和甘露糖含量两项质量指标之间存在一定的负相关性; 环境适应性好、叶绿素含量高的种质的质量较高。Zhang等^[13]分析30份铁皮石斛种质的农艺性状与多糖含量的关系, 发现茎长对多糖含量的直接影响最大, 直接通径系数1.568;节数、叶宽、平均节间距对多糖含量的间接影响较大, 间接通径系数分别是1.360、1.168、1.020。该研究提示, 选育铁皮石斛品种时, 通过考察茎长、节数、叶宽等农艺性状指标在一定程度上就能兼顾多糖含量指标。

1.2 铁皮石斛种质亲缘关系研究

Liu等^[14]分析云南、浙江、广西和广东4省36个铁皮石斛居群的农艺性状和化学成分的多样性, 发现同一铁皮石斛居群, 依据农艺性状的聚类结果与依据多糖和生物碱含量的聚类结果不一致, 证明了依据农艺性状和化学成分的含量差异来推断居群间的亲缘关系是不可靠的。目前, 绝大多数铁皮石斛种质的亲缘关系研究都是在分子水平开展的。研究发现, 铁皮石斛在物种水平有较高的遗传多样性, 但种群之间遗传多样性较低; 这主要是由于生境缺失或生境碎片化, 使铁皮石斛自然居群分布较分散, 居群之间的基因交流有限造成的^[15]。

铁皮石斛主要分布在我国浙江、安徽、江西、云南、广东和广西等省。铁皮石斛的地域分布与种质的亲缘关系是否存在相关性, 不同研究者报道的结果不一致, 目前尚无定论。Shen等^[16]利用RAMP标记将江西、浙江、广东、湖南、云南、贵州和广西7省的9个铁皮石斛野生居群分成3个类群, 亲缘关系与居群的地理分布存在相关性; Yuan等^[17]利用RAPD标记研究浙江省和云南省14个铁皮石斛人工栽培居群的亲缘关系, Dong等^[18]利用SSR标记研究安徽省和云南省11个铁皮石斛人工栽培居群的亲缘关系, 获得了类似的结果, 亲缘关系与居群地理种源显著相关。

Hou等^[19]通过对铁皮石斛及其4个近源种:黄石斛(D. tosaense)、曲茎石斛(D. ﬂexicaule)、滇桂石斛(D. scoriarum)及始兴石斛(D. shixingense), 合计34个居群叶绿体DNA和核DNA的谱系地理学研究, 认为云贵高原南部极有可能是这些物种的起源中心; 从云贵高原南部开始, 铁皮石斛依次向云贵高原东部、南岭山脉、武夷山脉、大别山脉和雁荡山脉不断扩张繁衍。Xu等^[20]利用SSR标记研究云南、浙江、广东、广西4省36个铁皮石斛人工栽培居群的结果表明, 亲缘关系与种源区域的联系并不紧密, 大多数来自不同区域的铁皮石斛被分在了同一类群中, 而相同种源地的铁皮石斛却被分在了不同的类群。

除了研究的技术方法、样本来源及数量不同外, 近年来铁皮石斛人工栽培范围扩大, 频繁的种质引进及交流加强了资源融合, 弱化了区域差异, 丰富了铁皮石斛种质的遗传多样性。与之相似, Ding等^[21]利用ISSR和RAPD标记研究广西、广东、湖南、浙江、云南、贵州和江西7省9个铁皮石斛自然居群, Li等^[22]利用AFLP标记研究广西、广东、湖南、浙江、云南、贵州、江西和福建8省12个铁皮石斛自然居群, 均发现居群间的遗传距离与居群的地理分布缺乏显著相关性。居群内个体间的遗传差异, 在前一项研究中占到总遗传变异性的78%以上, 在后一项研究中占到73%以上。研究人员推测是自然栖息地的破坏和人类的过度采挖导致了较严重的遗传漂移。

Bao等^[23]利用SRAP标记分析广东、云南、浙江、广西、安徽、贵州6省8个铁皮石斛自然分布居群的遗传多样性, 发现Nei总基因多样性系数(Ht)与Nei居群内基因多样性系数(Hs)差别较大, 不同居群铁皮石斛已经发生了一定程度的遗传分化, 居群内的遗传分化高于居群间的遗传分化。Cao等^[24]分析38份福建泰宁野生铁皮石斛叶绿体DNA rps16序列, 它们的种内K2P遗传距离为0~0.003 4, 5个多态性位点包括: 1位和29位的单个碱基缺失, 41位的单个碱基插入, 以及38位和136位的单个碱基颠换。Jiang等^[25]利用ISSR标记分析19份福建泰宁野生铁皮石斛种质和15份来自福建、浙江、云南等6省的栽培铁皮石斛种质的亲缘关系, 结果表明:泰宁野生铁皮石斛与人工栽培铁皮石斛的亲缘关系较远, 存在较大的遗传距离; 源自不同地理位置的野生种质倾向于聚集在独立的分支上; 依据地理来源将野生种质划分为4个种群, 与人工栽培种群相比, 野生种群具有更丰富的多样性和更高的种群分化系数。Zhou等^[26]利用SRAP标记研究黔西南州13份野生铁皮石斛种质与8份人工栽培种质的亲缘关系, 得出了类似的结果, 野生铁皮石斛遗传多样性水平比人工栽培的更高。为确保铁皮石斛种群的持续生存, 并维护种群的进化潜力, 迫切需要开展的工作包括:保护铁皮石斛生存环境, 保存种质, 提高种群的个体数量, 特别是提高遗传多样性丰富的种群的个体数量。

1.3 铁皮石斛种质鉴别的分子标记研究

利用分子标记鉴别植物物种是近年来发展较快的一项技术, 其用于区分石斛属植物的研究报道较多。Ding等^[27]研究认为rDNA ITS区因具备在细胞核中高度重复、不易被降解、种间信息位点丰富并且稳定等特点, 可被成功用于枫斗类石斛的种间鉴别。Jin等^[28]报道了一条全长302 bp的铁皮石斛特异性片段DS-302, 它作为SCAR标记, 能够快速、准确地区分铁皮石斛与细茎石斛(D. moniliforme)、霍山石斛(D. huoshanense)等10个同属近似物种。Dong等^[29]报道ITS序列60位和417位是铁皮石斛特有的SNP位点, 利用它们设计的铁皮石斛双阻滞位点特异性PCR引物, 能够特异性地鉴别铁皮石斛及其近源物种。利用DNA条形码技术鉴定石斛属植物的研究中, 推荐作为石斛属候选条形码的序列有:叶绿体基因组matK^[30]、psbA-trnH^[31]、petA-psbJ^[32]和psbK-psbI^[33]序列, 以及rDNA ITS^[34]和ITS2序列^[35]等。这类研究同时发现, 与利用单一序列相比, 利用组合序列可以提高石斛属植物的鉴别率。已报道的可以作为鉴别石斛属植物及其混伪品的DNA条形码序列组合包括: ITS+matK^{[36, 37]}和psbA-trnH+ 5S rRNA^[38]等。

利用分子标记技术开展的铁皮石斛种质资源研究也取得了一些有意义的结果。Ding等^[39]利用RAPD分子标记技术研究湖南、浙江、云南、福建等6省区8个铁皮石斛野生居群的遗传多样性, 共筛选出10个有效引物, 其中1条引物能有效区分8个野生居群, 且结果重现性较好。Cao等^[24]研究了50份不同来源铁皮石斛种质, 提出叶绿体DNA rps16序列可以作为研究铁皮石斛种内变异的DNA条形码序列之一, rps16-F: GGTGTAGATATGATCGAAAT, rps16-R: CCGATAAAGAATCAAACTTA。Hu等^[40]利用15份铁皮石斛种质资源筛选出4对可以用于铁皮石斛种质资源遗传多样性分析和指纹图谱构建的EST-SSR引物, 其中DN4-F: GGAGACATCGAAGAA GACAAGC, DN4-R: AAGAGTAAAATGCCACTGC ACA; DN10-F: TCTCTCACCAGAAAACAAAGCA, DN10-R: AAGAGTAAAATGCCACTGCACA; DN39-F: TTTACGACTGCCCTACCTTCAT, DN39-R: ACTCTT GTTTCCGAGAGATTGC; DN105-F: GGTTCGCCA TTCCAATATCTTA, DN105-R: ACCCACTGCAACT CTTCTCAAT。Fu等^[32]利用叶绿体DNA条形码技术研究15份不同采集地的铁皮石斛种质资源, 认为核心条形码matK + rbcL搭配petA-psbJ是适合铁皮石斛种质资源分子鉴定的条形码组合; 其中matK-F: CCC RTYCATCTGGAAATCTTGGTT, matK-R: GCTRTRA TAATGAGAAAGATTTCTGC; rbcL-1F: ATGTCACC ACAAACAGAAAC, rbcL-724R: TCGCATGTACCTG CAGTAGC; petA: AACARTTYGARAAGGTTCAATT, psbJ: ATAGGTACTGTARCYGGTATT。Ding等^[41]基于铁皮石斛ITS序列80位和132位的SNP位点设计了2对等位基因特异性PCR (AS-PCR, allele-specific PCR)引物, 能够成功鉴别广东韶关和福建顺昌两个自然居群的样品; 其中JB01-F: 5-CTTGCTGCTGAG ATGAAATCCACTGA-3, JB01-R: 5-TTATTGATATGC TTAAACTCAGCGGG-3; JB02-F: 5-GTGTGGACGT GATGAAGGATGGATGG-3, JB02-R: 5-TTATTGATA TGCTTAAACTCAGCGGG-3。

2 铁皮石斛遗传育种研究

收起

铁皮石斛在野外条件下的自然结实率不到5%, 生产上以人工异花授粉结实为主, 结实率大于90%^[42]。铁皮石斛的人工栽培历史不过三四十年, 目前广泛采用无菌播种的方式快速繁殖, 种质的优劣直接影响中药的品质与疗效。由于居群内个体间的遗传分化度较高, 加之栽培过程中忽视选育, 铁皮石斛已产生了多种遗传变异类型, 这是导致其栽培整齐度差、产量低、品质不稳定的主要原因。现有的种质资源研究结果表明, 铁皮石斛有着丰富的遗传多样性, 适应环境变化的能力强, 这预示出铁皮石斛育种和遗传改良的巨大潜力。

铁皮石斛良种选育和遗传育种工作起步晚, “天斛1号”是2006年由浙江省农作物品种审定委员会审定并发布的第一个品种^[43]。铁皮石斛品种选育初期, 产量是唯一的育种目标。对铁皮石斛农艺性状, 特别是与产量相关的农艺性状的研究, 如单株鲜条重、茎长、茎粗等, 能够直接为选育高产种质提供参考^[44]。随着种植范围和种植面积的扩大, 对化学、药理学等基础研究的深入, 以及药材质量标准的提高, 提升药材品质成为铁皮石斛品种选育的一个重要目标。但是目前在表型性状及化学成分调控机制的遗传学研究方面, 铁皮石斛仍然几乎是空白。

浙江省是最早开展铁皮石斛人工栽培和遗传育种研究的省份。尽管设施化栽培的普及和人工栽培技术的成熟使得铁皮石斛的栽培范围不断扩大, 但其种植地区仍以我国南方为主。本文归纳了现有的17个种源明确、特征特性描述详细的铁皮石斛品种的基本信息(表 1)^{[43, 45-66]}, 其中70%以上的品种出自广东和浙江2省, 它们大多具有产量高、多糖含量高等优势。铁皮石斛遗传育种的方法主要有:选择育种、杂交育种、诱变育种、生物技术育种等。现有铁皮石斛品种都是通过传统的选择育种和杂交育种获得的。尽管这两种方法存在周期长、效率低的缺点, 但它们仍是目前应用最有效的遗传育种方法。

2.1 选择育种

选择育种是从现有种质资源群体中选出具有优良性状的自然变异个体, 再使其繁育后代而获得新品种的方法。有助于提高选择效率的手段包括:利用科学而先进的选择方法确定适宜的种质群体; 全面评价育种基础材料的遗传性状, 确定各性状的遗传力和配合力; 利用适宜的光照和温度加快世代进度; 以及利用分子标记辅助育种等。长期以来, 铁皮石斛以野生居群生长, 个体之间的遗传性状差异大, 通过系统选育的方法, 可以从中获得具有高产、优质、抗病性强等优良性状的个体。表 1中有14个铁皮石斛品种是通过选择育种的方法获得的。

Chen等^[67]收集了12个不同种源的铁皮石斛种质, 分别从形态、产量、适应性3个方面选取了14个评价指标, 利用层次分析法筛选出适宜广西地区种植的品种, 并建立起铁皮石斛种质的综合评价体系。该系统中, 适应性是铁皮石斛种质资源评价和选育的最重要因子, 其次为形态因子; 筛选出的铁皮石斛种质资源评价指标, 按其重要性由高到低排序依次为:抗病性、管理难度、口感、萌芽能力和单条鲜重。

2.2 杂交育种

杂交育种是利用性状不同的父本与母本进行杂交以产生基因重组的后代, 然后根据育种目标选择出性状符合要求的后代, 再进行繁殖以获得新品种的方法。杂交育种可以将双亲本的控制不同性状的优良基因结合于一体, 也可以将双亲本中控制同一性状的不同微效基因积累起来, 产生在该性状上超过亲本的子代。正确选择亲本并予以合理组配是杂交育种的关键。石斛属植物种内杂交和种间杂交都有较好的亲和性, 因此杂交育种是该属植物育种的主要方法^{[5, 68]}。铁皮石斛人工异花授粉结实率高, 杂交育种是创新铁皮石斛种质资源, 提高其产量、质量及抗逆性的重要手段。表 1中的仙斛3号、双晖1号和雁吹雪3号是通过杂交育种方式获得的铁皮石斛品种。

Liu等^[69]通过系统分析铁皮石斛4个种源26个亲本的26个交配组合F1代的12个农艺性状发现, 全同胞F1代家系的植株高于半同胞F1代家系, 全同胞F1代家系内主要性状较半同胞F1代稳定, 提示在利用铁皮石斛杂交优势时, 开展对全同胞F1代的选择与利用会获得更高的遗传增益和更好的家系内一致性; 经主成分分析, 第1主成分的变异来源为叶面积、叶长和叶宽等叶片相关性状, 变异贡献率占38.18%;叶片宽的铁皮石斛在苗期表现出茎中部粗壮、苗高、分支少等较好的茎部性状。Zhang等^[70]测定11个铁皮石斛F1代的多糖含量后发现, 多数多糖含量低的亲本通过与多糖含量高的亲本杂交, 可以显著提高子代多糖含量, 杂交优势明显; Zhang等^[71]分析13个铁皮石斛F1代全同胞家系农艺性状与产量的相关性, 发现株高、茎粗、茎节数量、节间距、叶片数量、叶片长度、叶片宽度与产量成显著正相关, 萌蘖数小于4.5株/丛时与产量呈显著正相关; 逐步回归分析结果表明, 茎粗、叶片长度和叶片数量是选育高产铁皮石斛品种时应重点关注的农艺性状; 萌蘖数、茎粗和叶形指数3个性状在植株定植当年就比较稳定, 可用于早期选择。

2.3 诱变育种

诱变育种是指用物理或化学因素诱导植物的遗传特性发生变异, 再根据育种目标从变异群体中选择单株或个体, 进而培育成新的品种或种质的育种方法。诱变育种是继选择育种和杂交育种之后发展起来的一项现代育种技术。有益突变频率较低, 变异方向和性质难以控制, 是目前诱变育种面临的主要问题。

物理诱变是利用物理因素诱发生物体DNA发生变异, 其中应用最多的是辐射诱变。Xie等^[72]报道, γ射线照射可以使铁皮石斛原球茎DNA发生变异; SSR和ISSR分子标记分析, 辐照后差异条带比率1.01%~2.53%, 变异率与辐射剂量存在一定正相关; 但γ射线照射对铁皮石斛多糖含量没有显著影响, 经1.0 Gy·min^-1剂量率下50 Gy~110 Gy照射的铁皮石斛原球茎, 再经过10个月的继代、分化培养, 辐照幼苗茎的多糖含量与未经辐照幼苗茎的没有显著差异。空间诱变是我国上世纪80年代开始的一种物理诱变育种方法, 它是将种子送到太空, 利用太空中高真空、微重力、高辐射、弱地磁等特殊条件及其综合效应使种子发生基因变异, 再返回地面播种, 选择、培育获得新品种。“仙斛3号”铁皮石斛是以“仙斛1号”为父本, “514号”为母本, 先杂交得到F₁代蒴果, 再对F₁代蒴果进行航天搭载, 最后对诱变蒴果进行人工筛选获得的一个新品种^[50]。

化学诱变是利用化学诱变剂处理植物材料来诱导遗传物质发生突变。在铁皮石斛育种研究中应用较多的是秋水仙素倍性育种。植物中普遍存在多倍体现象。多倍体植物具有器官加大, 环境适应性较强, 抗逆性提高等优势。铁皮石斛体细胞染色体为二倍体, 2n = 38。Zhan等^[73]用秋水仙素处理铁皮石斛种胚原球茎, 获得了茎短粗、分支, 叶片小而厚的4倍体植株, 变异率最高达到85%。Zhang等^[74]用秋水仙素诱导铁皮石斛丛生芽, 获得茎增粗, 叶片增厚、颜色加深, 气孔保卫细胞增大、数量减少的4倍体植株, 最高变异率达48%。由体细胞诱变获得的无性多倍体植株, 其后代存在变异率低, 同质性增加, 易出现嵌合体及混倍体的不足。2n配子, 又称为未减数配子, 是自然界植物产生多倍体的主要途径。利用2n配子育种, 可以大大提高后代变异率, 加速对良种后代的选育^[75]。Wang等^[76]用秋水仙素处理铁皮石斛花蕾, 诱导获得了2n花粉, 最高诱导率达6.22%, 为创新铁皮石斛有性多倍化种质提供了参考。

2.4 生物育种

生物育种技术突破了传统育种技术中育种周期长、过程不可预测的局限, 能够提高育种的精确性、高效性、可控性和可预见性, 并为加快兰科植物的发育速度和种质的快速识别与鉴定提供了新的可能性^{[77, 78]}。生物育种技术主要包括:转基因育种、分子标记辅助选择育种、分子设计育种和智能不育杂交育种等。前两种生物育种技术已经用于铁皮石斛的育种研究。

转基因育种又被称为遗传修饰育种, 是利用基因工程技术, 将体外重组后的外源基因导入受体细胞, 通过外源基因在受体细胞内表达, 筛选出符合要求的新品种。与传统育种方法相比, 转基因育种可以针对目标性状精确改良, 且不受种属限制, 充分利用遗传资源, 但存在引起生态危机的风险, 且技术难度大。目前报道的铁皮石斛转基因育种方法主要包括:基因枪法、农杆菌介导法和花粉管通道法。

基因枪法是通过动力系统将带有基因的金属颗粒以一定速度射进靶细胞, 实现基因的稳定转化。该方法操作简单, 安全性高, 应用范围较广, 可一次处理多个细胞, 缺点是转化效率较低。Yang等^[79]用基因枪法将大麦抗旱耐盐基因lea3导入铁皮石斛原球茎中, 获得的转基因植株的耐盐胁迫能力显著增强。该研究报道, 采取以对数生长期前期的原球茎为转化受体, 转化前进行光照预分化处理, 以及轰击后进行过度恢复培养等措施后能显著提高抗性原球茎的得率; 尽管如此, 获得抗性再生植株的转化率仍比较低, 仅1%左右。

农杆菌介导法主要以植物的分生组织或生殖器官作为外源基因导入的受体, 通过真空渗透、浸蘸、注射等方法使农杆菌与受体材料接触完成可遗传细胞的转化。拷贝数低, 重复性好, 较少发生基因沉默现象, 转育周期较短, 能转化较大片段是农杆菌介导法的独特优势。为促进人工栽培铁皮石斛营养生长, 减少开花造成的养分消耗, 提高药材产量和品质, Wu等^[80]用农杆菌介导1-氨基环氧烷-1-羧酸(ACC)氧化酶的反义基因进入铁皮石斛原球茎细胞, 以抑制ACC氧化酶基因的表达, 减少植物体内乙烯的合成, 从而调控植物的生长发育过程。该研究中最高转化率为32.50%。

花粉管通道法是以植物授粉后形成的花粉管为通道, 使外源基因转化植物的受精卵或精子、卵子细胞而将其整合到受体基因组中。这种方法不受受体物种和基因型, 以及目的基因来源等因素的限制, 也无需诱导再生等人工培养过程, 是一种操作简单、快速高效的转基因育种方法。Xian等^[81]的研究发现, 携带GFP和GUS基因的质粒和农杆菌均可以作为载体, 通过花粉管通道法对铁皮石斛进行遗传转化; 以质粒和农杆菌为载体获得的结实率分别为62.3%和58.6%;经计算, 含GFP和GUS基因质粒处理的种子的最高转化率分别为7.8%和6.0%, 含GFP和GUS基因质粒的农杆菌处理的种子的最高转化率分别为5.0%和1.5%;与农杆菌相比, 质粒为载体的转化效果更好。

分子标记辅助育种是利用与目标性状紧密连锁的分子标记, 或目标性状基因序列本身的分子标记, 通过检测分子标记即可检测到目的基因的存在, 以达到选择目标性状的目的。分子标记辅助育种具有快速、准确, 不受环境和季节条件限制, 不受基因表达影响的优点^[82]。发现与目标基因紧密连锁的分子标记是提高育种效率的基础。RFLP、SSR、SNP等随机分子标记已经被广泛用于铁皮石斛种质资源研究。

2009年由Collard和Mackill^[83]开发的来源保守DNA序列多态性(conserved DNA-derived polymorphism, CDDP)是基于单引物扩增的一种目的分子标记技术, 它针对植物基因家族中的保守氨基酸序列设计引物, 能产生与目标性状连锁的功能性分子标记, 并且在不同物种间可以通用。植物的抗逆性往往不是由单一因子决定的, 如转录因子是通过调控一系列与逆境相关的功能基因的表达来提高植物的抗逆性。Wu等^[84]用针对转录因子WRKY, 生物与非生物因素胁迫应答相关基因MYB和在逆境胁迫中起调控作用的ERF转录因子设计的引物, 从43份野生和栽培的铁皮石斛种质资源中初步筛选出8份具有潜在优良抗性的植株, 为铁皮石斛抗性育种提供参考。

多糖是中药铁皮石斛的主要有效成分。培育多糖含量高的品种, 是铁皮石斛品种选育的重要目标之一。研究与铁皮石斛品质相关的数量性状的遗传规律, 可以为改善数量性状的遗传操作创造条件, 提高目标数量性状优良基因选择的可能性、准确性和预见性。Lu等^[85]利用细茎石斛(母本)和铁皮石斛(父本)及它们的111个F₁代个体, 构建了包含8 573个特异性位点扩增片段标记、涵盖19个连锁群、较好覆盖石斛属基因组的高密度单核苷酸多态性(SNP)综合遗传图谱; 鉴定出与茎总多糖含量相关的5个数量性状位点(quantitative trait locus, QTL), 表型方差解释率9.2%~11.8%, 分布在3个基因连锁群上; 5个QTLs区域中共有19个SNPs候选标记, 通过比对铁皮石斛基因组数据, 确定3个QTLs区域中的8个SNPs标记定位在铁皮石斛的4个基因上, 基因编号为: Dendrobium_GLEAN_10032348、Dendrobium_ GLEAN_10047849、Dendrobium_GLEAN_10074377和Dendrobium_GLEAN_10120125。

3 展望

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根据笔者的不完全统计, 目前已经有20多个铁皮石斛品种, 其中绝大多数以“非主要农作物品种”的形式发布, 少数以“园艺植物新品种”的形式发布。发布至今, 这些品种都还没有被用于铁皮石斛的产业化种植, 说明它们的技术优势不明显, 缺乏经济价值。发展高产、优质、抗逆的优良品种是铁皮石斛规范化生产的必然趋势。对此, 当务之急是集中力量深入开展铁皮石斛遗传基础研究, 揭示遗传物质在铁皮石斛进化过程中流动、传递的本质, 阐明种内遗传关系, 构建铁皮石斛遗传连锁图谱, 为相关数量性状的基因定位和分离奠定基础, 进而揭示品种性状的遗传规律, 为选育优良品种提供理论依据。

规范化生产的目的是获得质量稳定的中药材, 优良品种是药材质量稳定的基础。因此, 质量指标应作为铁皮石斛品种选育的优先目标, 其次是产量及抗性指标。以此为原则, 根据铁皮石斛生物学特性, 利用系统、科学的方法筛选品种选育指标, 构建选育体系、评价体系及繁育体系。在产量与当前栽培品种相仿的前提下, 可以将多糖含量、多糖的单糖组成及比例、小分子特征性成分含量等与质量品质相关的指标, 以及抗当地1种以上主要病害或耐受当地主要不良环境因子等抗性指标纳入到铁皮石斛品种选育的目标中。

基金

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中国医学科学院医学与健康科技创新工程项目(2017-I2M-03-013)
国家自然科学基金资助项目(81573526)

参考文献

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文献

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2018年第53卷第9期

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doi: 10.16438/j.0513-4870.2018-0286

接收时间：2018-04-02
首发时间：2026-01-15
出版时间：2018-09-12

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收稿日期：2018-04-02
修回日期：2018-05-10

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中国医学科学院医学与健康科技创新工程项目(2017-I2M-03-013)

国家自然科学基金资助项目(81573526)

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中国医学科学院、北京协和医学院药用植物研究所, 北京 100193

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* 陈晓梅, Tel:86-10-57833241, E-mail:cxm_implad@163.com;

郭顺星, Tel:86-10-57833231, E-mail:sxguo1986@163.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Variety	Phenotypic characteristic	Average yield	Quality characteristic	Variety advantage	Source	Breeding method
Tianhu 1^[43]	SH 20-60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots	0.91 kg·m^-2 (20 months growth, fresh weight)	DR 15.5%, PC 15.6%	Good comprehensive characters, higher quality, better low- temperature and disease resistant	Domestication of wild resources	Selective breeding
Xianhu 1^[45]	SH 20-30 cm, up to 60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots	2.29 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 24.6%, PC 30.4%	Higher yield and PC, better adverse resistance	Domestication of wild resources Sxg-2	Selective breeding
Xianhu 2^{[46, 47]}	SH 30-50 cm, SD 6-10 mm, ID 15-25 mm, stem with purple spots; LL 4-7 cm, LW 1-2.5 cm, LT 0.6-0.9 mm, LS with purple spots	2.49 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 21.1%, PC 29.0%	Higher yield and PC, better low- temperature and disease resistant	Domestication of wild resources Xianzi 1	Selective breeding
Xianhu 3^[48]	SH 8-21 cm, SD 2-6 mm, ID 8-14 mm; LL 2-7 cm, LW 0.8-2.2 cm, edge and main vein of leaves often with purple spots	2.52 kg·m^-2 (24 months growth, fresh weight of stems and leaves)	DR 33.4%, EEC 13.85%, MC 21.75%, R_M/G 4.42	Higher yield, better adaptability and adverse resistance	MP: Xianhu1; FP: No. 514	Cross breeding and mutation breeding
Senshan 1^[49]	SH 14-36.6 cm, SD 3.58-6 mm; LL 3.8-5.3 cm, LW 1.7-2.2 cm, leaf back and LS often with purple spot	1.35 kg·m^-2 (2 years growth)	PC 24.45%	Higher yield, better adaptability	Domestication of wild resources collected from Yunnan	Selective breeding
Guihu 1^{[50, 51]}	SH 15-60 cm, SD 4-7.2 mm, stem with purple red spots/patches; LL 2-7 cm, LW 0.5-2.5 cm, leaf and LS with purple red spots/patches	0.23-0.64 kg·m^-2 (18-20 months growth, the first harvest); 0.38-0.76 kg·m^-2 (harvest once every 12 to 14 months after the first harvest, the second and third one)	PC 28.0%- 35.9% (fresh stems)	Better drought and root rot resistance	Selection of good individual from wild seedlings (Nazuo Miao Nationality Township, Xilin County, Guangxi)	Selective breeding
Baishishan 1^[52]	SH 12-65 cm, SD 4-8 mm, ID 1.5-2.5 cm, stem with purple spots; LL 4-6.5 cm, LW 1.3-2.5 cm, LT 7-9 mm, leaf edge often pale purple, leaf back often with pale purple spots	0.30-0.68 kg·m^-2 (18-24 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)	PC > 29% (dry stem)	/	Selection of good individual from test tube seedlings (purple stem of wild adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)	Selective breeding
Baishishan 2^[53]	SH 15-70 cm, SD 4-9 mm, ID 2-3 cm, no spots on stem; LL 4-6.5 cm, LW 1.2-2.5 cm, LT 7-9 mm	0.30-0.76 kg·m^-2 (18-20 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)	PC about 31%	/	Selection of good individual from wild seedlings (green stem of adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)	Selective breeding
Qinggu 1^[54]	SH 35-37 cm, SD 10.5-11.5 mm, stem full with purple spots; leaf edge purple, leaf back with purple spots	0.69 kg·m^-2 (fresh stems, 2 years growth) 0.72 kg·m^-2 (fresh stems, 3 years growth)	PC 47.7% (2 years growth); PC 47.2% (3 years growth)	/	Collection of wild resources from Yunnan; then selection good single-plant	Selective breeding
Zhongke 1^{[55, 56]}	SH up to 50 cm, SD up to 7 mm; leaves rectangular and lanceolate, LL 5-6 cm, LW 2-3 cm, LS with rust spots	1.5 kg·m^-2 (18 months growth of tissue cultured seedling)	PC about 30% (dry stems)	Higher yield, better high-temperature and disease resistance	T13 (Guangnan County, Yunnan), after multiple generations of self-breeding	Selective breeding
Shuanghui 1^[57-59]	SH 30-50 cm, SD 5-8 mm, ID 1.5-2.5 cm; leaves spindle, LL 6.0 cm, LW 2.0 cm	1.4 kg·m^-2 (fresh stem, 2 years growth)	PC 32.7%, MC 19.2%	Higher yield, better disease and adverse resistance	FP: red stem (Yandang Mountains, Zhejiang), MP: green stem (Guangnan County, Yunnan); artificial cross-pollination, single- plant selection of offspring, multiple generations of selfing purification	Cross breeding
Zhongke Congdu^{[60, 61]}	SH up to 55 cm and SD 6 mm after 2 years growth; leaves rectangular and lanceolate, LL 4-6 cm, LW 2-3 cm, LS with rust spots	1.2 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 32.1%, MC 23.7% (dry stem)	Higher PC, better adverse resistance	Introduction of the plants from Yachang orchid plant nature reserve (Leye County, Guangxi); then selection from multiple generations of self-breeding	Selective breeding
Zhongke Congdu 2^[62]	SH 34.3 cm and SD 6 mm after 2 years growth	1.36 kg·m^-2 (fresh stems, 18 months growth of tissue cultured seedling)	PC 42.4%, MC 29.4%, EEC 13.0%, R_M/G 2.6 (dry stem)	Higher PC, better disease resistance	Wild resources collected from Dao County, Hunan; selection after 4 generations of self-breeding	Selective breeding
Yanchuixue 3^{[62, 63]}	SH 25.5 cm and SD 6 mm after 2 years growth, stem with rust spots; LL 4.8 cm, LW 1.5 cm	0.62 kg·m^-2 (fresh stems)	PC 57.5%, MC32.6%, R_M/G 2.3 (dry stem)	Better disease resistance	Hybrid of green stem (Tiantai Mountains) and red stem (Yandang Mountains) plants	Cross breeding
Zhongke 3^[64]	SH 36.2 cm and SD 4.7 mm after 2 years growth, stem with rust spots; leaves narrow ovate, LL 5.1 cm, LW 1.6 cm	1.45 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 39.4%, MC 18.6%, EEC 6.6% (dry stem)	Better disease and adverse resistance	Wild resource T636 (Dao County, Hunan); selection after 4 generations of self-breeding	Selective breeding
Zhongke 4^[64]	SH 48.0 cm and SD 5 mm after 2 years growth, stem with rust spots; leaves ovate, LL 5.7 cm, LW 1.9 cm	1.56 kg·m^-2 (18 months growth of tissue cultured seedling)	PC 39.0%, MC 20.1%, EEC 6.6% (dry stem)	Better disease and adverse resistance	Wild resource T709 (Lang Mountains, Xinning County, Hunan); selection after 4 generations of self-breeding	Selective breeding
Fuhu 1^{[65, 66]}	SH 20.8 cm, SD 5.1 mm and ID 1.5 cm after 2 years growth, stem with pale purple spots; leaves long elliptic, LL 5 cm, LW 1.6 cm	0.48 kg·m^-2 (fresh stem)	PC 51.8%, MC 29.3%, R_M/G 2.5 (dry stem)	Higher yield and quality, lower incidence of black spot and anthracnose	Selection of wild resource L-2 from Guaizhai Mountains, Liancheng County, Fujian	Selective breeding

Variety

Phenotypic characteristic

Average yield

Quality characteristic

Variety advantage

Source

Breeding method

Tianhu 1^[43]

SH 20-60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots

0.91 kg·m^-2 (20 months growth, fresh weight)

DR 15.5%, PC 15.6%

Good comprehensive characters, higher quality, better low- temperature and disease resistant

Domestication of wild resources

Selective breeding

Xianhu 1^[45]

SH 20-30 cm, up to 60 cm, SD 5-10 mm; LL 4-7 cm, LW 1-2 cm, edge and main vein of leaves pale purple; LS with purple spots

2.29 kg·m^-2 (24 months growth, fresh weight of stems and leaves)

DR 24.6%, PC 30.4%

Higher yield and PC, better adverse resistance

Domestication of wild resources Sxg-2

Selective breeding

Xianhu 2^{[46, 47]}

SH 30-50 cm, SD 6-10 mm, ID 15-25 mm, stem with purple spots; LL 4-7 cm, LW 1-2.5 cm, LT 0.6-0.9 mm, LS with purple spots

2.49 kg·m^-2 (24 months growth, fresh weight of stems and leaves)

DR 21.1%, PC 29.0%

Higher yield and PC, better low- temperature and disease resistant

Domestication of wild resources Xianzi 1

Selective breeding

Xianhu 3^[48]

SH 8-21 cm, SD 2-6 mm, ID 8-14 mm; LL 2-7 cm, LW 0.8-2.2 cm, edge and main vein of leaves often with purple spots

2.52 kg·m^-2 (24 months growth, fresh weight of stems and leaves)

DR 33.4%, EEC 13.85%, MC 21.75%, R_M/G 4.42

Higher yield, better adaptability and adverse resistance

MP: Xianhu1; FP: No. 514

Cross breeding and mutation breeding

Senshan 1^[49]

SH 14-36.6 cm, SD 3.58-6 mm; LL 3.8-5.3 cm, LW 1.7-2.2 cm, leaf back and LS often with purple spot

1.35 kg·m^-2 (2 years growth)

PC 24.45%

Higher yield, better adaptability

Domestication of wild resources collected from Yunnan

Selective breeding

Guihu 1^{[50, 51]}

SH 15-60 cm, SD 4-7.2 mm, stem with purple red spots/patches; LL 2-7 cm, LW 0.5-2.5 cm, leaf and LS with purple red spots/patches

0.23-0.64 kg·m^-2 (18-20 months growth, the first harvest); 0.38-0.76 kg·m^-2 (harvest once every 12 to 14 months after the first harvest, the second and third one)

PC 28.0%- 35.9% (fresh stems)

Better drought and root rot resistance

Selection of good individual from wild seedlings (Nazuo Miao Nationality Township, Xilin County, Guangxi)

Selective breeding

Baishishan 1^[52]

SH 12-65 cm, SD 4-8 mm, ID 1.5-2.5 cm, stem with purple spots; LL 4-6.5 cm, LW 1.3-2.5 cm, LT 7-9 mm, leaf edge often pale purple, leaf back often with pale purple spots

0.30-0.68 kg·m^-2 (18-24 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)

PC > 29% (dry stem)

Selection of good individual from test tube seedlings (purple stem of wild adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)

Selective breeding

Baishishan 2^[53]

SH 15-70 cm, SD 4-9 mm, ID 2-3 cm, no spots on stem; LL 4-6.5 cm, LW 1.2-2.5 cm, LT 7-9 mm

0.30-0.76 kg·m^-2 (18-20 months growth, the first harvest); 0.45-1.52 kg·m^-2 (harvest once every 12 months after the first harvest, the second and third one)

PC about 31%

Selection of good individual from wild seedlings (green stem of adult plant, Baishi Mountains, Matong Township, Guiping City, Guangxi)

Selective breeding

Qinggu 1^[54]

SH 35-37 cm, SD 10.5-11.5 mm, stem full with purple spots; leaf edge purple, leaf back with purple spots

0.69 kg·m^-2 (fresh stems, 2 years growth) 0.72 kg·m^-2 (fresh stems, 3 years growth)

PC 47.7% (2 years growth); PC 47.2% (3 years growth)

Collection of wild resources from Yunnan; then selection good single-plant

Selective breeding

Zhongke 1^{[55, 56]}

SH up to 50 cm, SD up to 7 mm; leaves rectangular and lanceolate, LL 5-6 cm, LW 2-3 cm, LS with rust spots

1.5 kg·m^-2 (18 months growth of tissue cultured seedling)

PC about 30% (dry stems)

Higher yield, better high-temperature and disease resistance

T13 (Guangnan County, Yunnan), after multiple generations of self-breeding

Selective breeding

Shuanghui 1^[57-59]

SH 30-50 cm, SD 5-8 mm, ID 1.5-2.5 cm; leaves spindle, LL 6.0 cm, LW 2.0 cm

1.4 kg·m^-2 (fresh stem, 2 years growth)

PC 32.7%, MC 19.2%

Higher yield, better disease and adverse resistance

FP: red stem (Yandang Mountains, Zhejiang), MP: green stem (Guangnan County, Yunnan); artificial cross-pollination, single- plant selection of offspring, multiple generations of selfing purification

Cross breeding

Zhongke Congdu^{[60, 61]}

SH up to 55 cm and SD 6 mm after 2 years growth; leaves rectangular and lanceolate, LL 4-6 cm, LW 2-3 cm, LS with rust spots

1.2 kg·m^-2 (18 months growth of tissue cultured seedling)

PC 32.1%, MC 23.7% (dry stem)

Higher PC, better adverse resistance

Introduction of the plants from Yachang orchid plant nature reserve (Leye County, Guangxi); then selection from multiple generations of self-breeding

Selective breeding

Zhongke Congdu 2^[62]

SH 34.3 cm and SD 6 mm after 2 years growth

1.36 kg·m^-2 (fresh stems, 18 months growth of tissue cultured seedling)

PC 42.4%, MC 29.4%, EEC 13.0%, R_M/G 2.6 (dry stem)

Higher PC, better disease resistance

Wild resources collected from Dao County, Hunan; selection after 4 generations of self-breeding

Selective breeding

Yanchuixue 3^{[62, 63]}

SH 25.5 cm and SD 6 mm after 2 years growth, stem with rust spots; LL 4.8 cm, LW 1.5 cm

0.62 kg·m^-2 (fresh stems)

PC 57.5%, MC32.6%, R_M/G 2.3 (dry stem)

Better disease resistance

Hybrid of green stem (Tiantai Mountains) and red stem (Yandang Mountains) plants

Cross breeding

Zhongke 3^[64]

SH 36.2 cm and SD 4.7 mm after 2 years growth, stem with rust spots; leaves narrow ovate, LL 5.1 cm, LW 1.6 cm

1.45 kg·m^-2 (18 months growth of tissue cultured seedling)

PC 39.4%, MC 18.6%, EEC 6.6% (dry stem)

Better disease and adverse resistance

Wild resource T636 (Dao County, Hunan); selection after 4 generations of self-breeding

Selective breeding

Zhongke 4^[64]

SH 48.0 cm and SD 5 mm after 2 years growth, stem with rust spots; leaves ovate, LL 5.7 cm, LW 1.9 cm

1.56 kg·m^-2 (18 months growth of tissue cultured seedling)

PC 39.0%, MC 20.1%, EEC 6.6% (dry stem)

Better disease and adverse resistance

Wild resource T709 (Lang Mountains, Xinning County, Hunan); selection after 4 generations of self-breeding

Selective breeding

Fuhu 1^{[65, 66]}

SH 20.8 cm, SD 5.1 mm and ID 1.5 cm after 2 years growth, stem with pale purple spots; leaves long elliptic, LL 5 cm, LW 1.6 cm

0.48 kg·m^-2 (fresh stem)

PC 51.8%, MC 29.3%, R_M/G 2.5 (dry stem)

Higher yield and quality, lower incidence of black spot and anthracnose

Selection of wild resource L-2 from Guaizhai Mountains, Liancheng County, Fujian

Selective breeding