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2020 Volume 42 Issue 12
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LI Qiao-ling, XIAO Bo, DENG Cai-fu, et al. Collection, Preservation, Evaluation and Optimization of Germplasm Resources of Gastrodia in Its Main Producing Areas[J]. Journal of Southwest University Natural Science Edition, 2020, 42(12): 54-59. doi: 10.13718/j.cnki.xdzk.2020.12.006
Citation: LI Qiao-ling, XIAO Bo, DENG Cai-fu, et al. Collection, Preservation, Evaluation and Optimization of Germplasm Resources of Gastrodia in Its Main Producing Areas[J]. Journal of Southwest University Natural Science Edition, 2020, 42(12): 54-59. doi: 10.13718/j.cnki.xdzk.2020.12.006
shu

Collection, Preservation, Evaluation and Optimization of Germplasm Resources of Gastrodia in Its Main Producing Areas

  • The ResearchInstitute of Medicine Plantation of Chongqing/Chongqing Centerforthe Selection and Evaluation of Traditional Chinese Medicine Breeding , Nanchuan Chongqing 408435 , China

More Information
  • Corresponding author: YANG Yi
  • Received Date: 28/10/2019
    Available Online: 20/12/2020

  • MSC: S567.2

  • Gastrodia elata is a traditional Chinese herbal medicine with remarkable medicinal effects. However, a great variety of Gastrodia germplasms differ considerably in their appearance and production performance, on the one hand, and in the intrinsic quality of the medicinal materials, on the other. Therefore, it is of great significance to screen out the excellent Gastrodia germplasm resources and provide basic materials for the Chinese Gastrodia germplasm resource database and new variety breeding. In a study reported herein, 27 Gastrodia germplasms were collected from the main Gastrodia production areas of China, and they were introduced to the Dafoyan (Lecun Forest Farm) in Nanchuan of Chongqing. After sexual reproduction, the Gastrodia tubers were harvested at maturity, and the gastrodin and p-hydroxybenzyl alcohol were extracted by the ultrasonic method and chromatography, and their contents were determined by ultraviolet spectrophotometry. The results indicated that Gastrodia germplasms of different origins differed significantly in their appearance traits and yield and in the contents of gastrodin and p-hydroxybenzyl alcohol. Of the 27 Gastrodia germplasms studied, YX from Yuexi of Anhui and CG from Chenggu of Shaanxi showed outstanding performance in terms of traits, yield and active medicinal components. The average fresh weight, average dry weight, yield and the total content of gastrodin and p-hydroxybenzyl alcohol were 184.87±9.79 g, 35.97±3.52 g, 6.53±0.41 kg/m2 and (0.468±0.05)% for YX, and 157.64±11.31 g, 26.08±2.24 g, 5.74±0.76 kg/m2 and (0.565±0.08)% for CG. In conclusion, the quality and yield of YX and CG were significantly better than those of the other germplasms. Their total contents of gastrodin and p-hydroxybenzyl alcohol were higher than 0.25% (Pharmacopoeia standard), and their theoretical yield was above 3 500 kg/mu (15 mu=1 hectare). Extension of the cultivation of these two excellent varieties is one of the most effective ways to improve the quality of Gastrodia in Chongqing.
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Collection, Preservation, Evaluation and Optimization of Germplasm Resources of Gastrodia in Its Main Producing Areas

    Corresponding author: YANG Yi
  • Received Date: 28/10/2019
  • Available Online: 20/12/2020

Abstract: Gastrodia elata is a traditional Chinese herbal medicine with remarkable medicinal effects. However, a great variety of Gastrodia germplasms differ considerably in their appearance and production performance, on the one hand, and in the intrinsic quality of the medicinal materials, on the other. Therefore, it is of great significance to screen out the excellent Gastrodia germplasm resources and provide basic materials for the Chinese Gastrodia germplasm resource database and new variety breeding. In a study reported herein, 27 Gastrodia germplasms were collected from the main Gastrodia production areas of China, and they were introduced to the Dafoyan (Lecun Forest Farm) in Nanchuan of Chongqing. After sexual reproduction, the Gastrodia tubers were harvested at maturity, and the gastrodin and p-hydroxybenzyl alcohol were extracted by the ultrasonic method and chromatography, and their contents were determined by ultraviolet spectrophotometry. The results indicated that Gastrodia germplasms of different origins differed significantly in their appearance traits and yield and in the contents of gastrodin and p-hydroxybenzyl alcohol. Of the 27 Gastrodia germplasms studied, YX from Yuexi of Anhui and CG from Chenggu of Shaanxi showed outstanding performance in terms of traits, yield and active medicinal components. The average fresh weight, average dry weight, yield and the total content of gastrodin and p-hydroxybenzyl alcohol were 184.87±9.79 g, 35.97±3.52 g, 6.53±0.41 kg/m2 and (0.468±0.05)% for YX, and 157.64±11.31 g, 26.08±2.24 g, 5.74±0.76 kg/m2 and (0.565±0.08)% for CG. In conclusion, the quality and yield of YX and CG were significantly better than those of the other germplasms. Their total contents of gastrodin and p-hydroxybenzyl alcohol were higher than 0.25% (Pharmacopoeia standard), and their theoretical yield was above 3 500 kg/mu (15 mu=1 hectare). Extension of the cultivation of these two excellent varieties is one of the most effective ways to improve the quality of Gastrodia in Chongqing.

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  • 中药材种质资源的研究是提高中药质量的关键和源头，种质的优劣是影响中药材产量和质量的重要因素，而保证中药材优质是中药产业发展的根本措施.对中药材种质资源进行调查、收集与评价，筛选优质中药材种质资源，有利于中药材品种的品质改良，进而从源头上提高我国中药产品质量和中药材生产技术水平[1].

    天麻为我国传统名贵中药材，药用功效显著，在《神农本草经》和《本草纲目》里，均有天麻“除百病益寿延年”的记载[2].天麻具有祛风湿、止痛、行气活血之功效，主要用于治疗头痛眩晕、肢体麻木、癫痫及心脑血管疾病等[3-4].近年来，天麻除了传统入药外，也被广泛用于食品、药品的开发，市场需求量极大[5].由于天麻有着较为明显的经济效益优势，其种植区域大、种植种类多，通过自然和人为杂交方式也培育出了许多不同的地方品种.但众多天麻品种一方面在药材外观性状、生产性能等方面存在差异，另一方面药材内在质量也存在差别[6].因此，筛选出优质高产的天麻种质资源成为提高天麻品质和经济效益的必要途径.

    本研究将收集到的来自湖北、陕西、重庆、云南等不同产地的天麻种质资源27份，引种到南川区水江镇辉煌村2组大佛岩乐村林场建立种质资源圃.通过实地的有性生产繁殖，比较不同产地天麻种质资源生产性能、经济性状差异.依据《中国药典》2015年版，以天麻浸出物、灰分、天麻素和对羟基苯甲醇为品质研究指标，评价各天麻种质的品质特性.综合产量与品质特性，对不同产地天麻种质资源进行综合评价，筛选出经济性状较优的天麻品种，为重庆地区天麻优良品种的选育及规范化栽培打下前期基础.

1.   实验地概况

  • 实验地位于重庆市南川区水江镇辉煌村2组大佛岩乐村林场，地理坐标为北纬29°15′47.79″，东经107°21′3.45″.森林覆盖率达90%以上，立体海拔在1 050~1 835 m之间.该地属于亚热带湿润季风气候，具有冬短、春早、夏长、雨热同季、气候垂直变化明显的特点，年降雨量在1 250 mm左右，雾日260 d左右，年均气温12.5~14.5 ℃，年均无霜期270~287 d.土壤肥沃，以山地黄壤土和腐殖土为主[7].

2.   材料与方法

    2.1.   材料

  • 2016年2-3月，分别从全国天麻主产区湖北省宜昌市下堡坪乡、陕西省西乡县城关镇、陕西省城固县五堵镇、陕西省宁强县汉源镇、四川省青川县茶坝乡、安徽省岳西县巍岭乡、云南省昭通市彝良县小草坝乡、四川省绵阳市江油市大康镇、重庆市南川区水江镇乐村林场等共9个样地收集天麻种质资源(表 1)，每个样地分别收集3份，共计27份天麻种质资源.所有样品经重庆市药物种植研究所研究员邓才富鉴定为兰科植物天麻(Gastrodia elata Bl.).

    天麻有性繁殖方法具体参照张志明等[8].将引种回来的箭麻于3月上旬栽植，4月开花时采取人工授粉.授粉后约15~20 d左右，蒴果成熟采收.将采收的成熟蒴果同萌发菌石斛小菇共同播种，繁育不同原产地的零代天麻.次年将零代天麻米麻按照700 g/m2的密度播种，每个品种3个重复，随机布置.每个点布置面积为1.3 m2，产量测定时全部采收称量.

    • 2.2.   实验方法

    2.2.1.   天麻外观性状及产量测定

  • 天麻外观性状测定：将收集到的9个不同产区天麻种质资源经过有性繁殖后，次年12月采收天麻块茎进行测量.每组随机抽取20个天麻样品，分别测定其长度、直径、直径/长度、鲜质量和干质量.

    产量测定：测定单位面积天麻的鲜质量和干质量.

    • 2.2.2.   不同天麻种质资源成分含量测定

  • 新鲜天麻洗净泥沙后，采用YEN50液体真空浓缩煎药机蒸汽蒸制20 min，50 ℃烘箱干燥1夜切厚片2~4 mm，继续50 ℃烘干，粉碎过筛放冰箱备用，按《中国药典》2015年版一部天麻标准规定测定其总灰分、浸出物、有效成分(天麻素、对羟基苯甲醇).天麻素对照品和对羟基苯甲醇对照品均购自中国食品药品检定研究院，批号分别为110807-201608，111970-201501.

    • 2.2.3.   数据处理

  • 用Microsoft Excel 2010软件计算各外观性状平均值和标准差，用SPSS 20.0软件进行方差分析.

3.   实验结果

    3.1.   天麻外观性状

  • 影响天麻经济价值因素除了市场价格外，主要为质量和产量.天麻鲜品的市场价格与其大小息息相关，大小不同则价格差异巨大，而鲜天麻的大小又与其农艺性状密切相关[9]. 9个产区的天麻成熟块茎性状的统计结果(表 2)显示：9份天麻材料中以安徽岳西的种源直径、直径/长度、鲜质量、干质量最大，分别为6.25 cm，0.56，184.87 g和35.97 g，陕西城固和四川青川的次之; 湖北宜昌种源的直径、直径/长度、鲜质量、干质量最小，分别为3.05 cm，0.26，70.19 g和11.12 g，但其长度值最大，为细长类型.而重庆南川本地天麻从单株产量方面讲，处于居中位置. 9份天麻种质资源在重庆南川地区栽培后，直径、直径/长度、鲜质量、干质量差异有统计学意义，这些差异可能是由各种质资源的长期遗传变异及栽培地地理环境因素造成的[10].

    • 3.2.   天麻产量测定

  • 将引种来的9份天麻种质资源通过实地产量测定，结果表明不同种源天麻产量差异存在统计学意义，最高的安徽岳西天麻样品产量为6.53 kg/m2，最低的陕西宁强天麻样品产量为2.69 kg/m2，两者相差2.43倍，不同种源间单位面积鲜质量产量经方差分析差异有统计学意义(表 3).

    • 3.3.   含量测定

  • 《中国药典》2015年版中天麻项下规定了天麻的水分≤15.0%，总灰分≤4.5%，醇浸出物≥15.0%. 9个天麻样品中，水分介于4.65%~10.24%之间，均符合《中国药典》标准; 总灰分介于2.01%~3.24%之间，均值2.53%，相对标准偏差(RSD)为17.61%，均低于《中国药典》规定的4.5%标准; 稀乙醇热浸出物介于20.33%~33.17%，均值27.13%，RSD为14.11%，均远高于《中国药典》规定的15.0%标准. 3个样品YC，NQ和NC为《中国药典》标准规定的2倍以上，达到30.0%以上.各产地所采收的天麻样品在水分、总灰分和浸出物方面都达到了《中国药典》标准(表 4).

    9个天麻主产区的天麻样品中天麻素、对羟基苯甲醇及二者总质量分数测定结果(表 5)显示，不同产地天麻的天麻素质量分数差异有统计学意义，天麻素质量分数均值为0.283%(0.157%~0.381%)，最高的安徽岳西种源是最低的湖北宜昌种源的2.43倍; 对羟基苯甲醇质量分数介于0.067%~0.191%之间，均值0.126%，以陕西城固和云南彝良的种源最优，分别是陕西西乡种源的2.85倍和2.78倍; 不同种源的天麻素和对羟基苯甲醇二者总质量分数均符合《中国药典》规定的0.25%标准，以陕西城固种源的总质量分数最高，四川江油和云南彝良、安徽岳西的种源次之.因此，陕西城固、安徽岳西等优质种源的开发利用，对于当前提高重庆地区天麻质量具有重要的现实意义.

4.   讨论

  • 中药材大多来源于药用植物，由于人为的过度采挖，我国野生药用植物资源不断减少，有的甚至濒临灭绝，目前许多中药材的来源多依赖于人工种植.在中药材的人工种植过程中，如何保证质量和提高产量已经成为生产中的关键问题[11].种质是决定中药材质量与产量的关键因素，虽然中药材的种植品种不断增加、面积逐年扩大，但有关药用植物种质资源方面的研究工作仍相对滞后[12].主要体现在如下几个方面：许多地道中药材的种质混乱，缺乏系统的搜集和整理; 中药材优良种质的选育进展缓慢，生产中缺乏具有推广价值的优良品种; 缺乏合理的品种规范化留种技术，部分中药材出现种质退化现象.以上存在的问题是中药材在人工种植过程中产量低、质量差的主要原因，也是实现中药材生产规范化、规模化与产业化的重要障碍[13].对现有的中药材种质资源进行系统的调查、收集、整理与评价，选育优良种质并推广应用，是保证中药资源及其产业可持续发展的重要基础.

    本文比较分析了我国天麻主产区9个种源的外观性状、产量和内在化学成分.从天麻外观性状来看，不同产地天麻块根在长度、直径、单株鲜质量、干质量等方面差异有统计学意义，其中以安徽岳西的种源直径、直径/长度、鲜质量、干质量最优，陕西城固和四川青川的种源次之; 而重庆南川本地天麻从单株鲜质量方面讲，处于居中位置.不同产区天麻种源产量结果显示，安徽岳西天麻样品产量最高，为6.53 kg/m2，是陕西宁强种源的2.43倍.从化学成分来看，结合现行2015版《中国药典》天麻标准，各产区的天麻样品的总灰分和稀乙醇浸出物均符合且远优于《中国药典》规定标准; 天麻素和对羟基苯甲醇两个活性成分的总质量分数均符合《中国药典》规定标准，且多数样品远高于《中国药典》标准，相关质量指标测试结果都较理想.综合相关质量、产量及外观性状指标，陕西城固和安徽岳西2个种源天麻明显优于其他种源.因此，在重庆及其类似的天麻产区推广应用这2个优良种源，是现有技术条件下提高天麻质量的最有效途径.

    从2018年社会经济发展状况及天麻市场行情来看，种植天麻具有较好的经济效益和一定的社会效益.天麻的市场容量有限，生产技术相对成熟，在天麻菌材资源丰富、气候适宜的地方，可合理间伐菌材，适度发展天麻产业[14-15].然而，天麻产业的可持续发展同样面临野生资源破坏、种质退化等问题[16]，所以我们对天麻野生资源应坚持开发与保护并重、及时选育优良品种进行推广以及合理改进栽培措施以促进天麻产业的健康可持续发展.

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