川芎苓种标准化及种质保存技术的研究
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摘要
川芎为伞形科植物川芎Ligusticum chuanxiong Hort的根茎,为我国的传统中药,也是著名的川产道地药材,具有近两千年的栽培和使用历史,长期以来生产上均以无性方式进行繁殖,其繁殖材料为膨大的地上茎节(苓种)。经过上千年的栽种,加上缺乏选种育种,存在一定程度的种质退化。四川主产区药农对苓种繁育仍然停留在自繁自用,缺乏统一的生产技术规程和科学的组织管理,且没有苓种分级标准和检验规程,导致川芎苓种质量良莠不齐。另外,目前生产上仍然采用传统的自然存放方法贮存苓种,不能久贮；种质保存只能采用传统的田间保存方法,存在耗费人力物力,容易损失等诸多缺点,而相关研究尚属空白。
中药材质量稳定需要中药材生产的规范化,中药材生产规范化需要中药材种子种苗的标准化。优良品种和优质的种子种苗是实现中药材规范化生产的基础和源头。本文针对川芎规范化生产中的薄弱环节,以苓种标准化问题为切入点,开展了川芎苓种的标准化研究和种质保存技术研究,以推动川芎苓种的标准化进程,为川芎规范化生产提供源头保障。以生产出优质、高产、稳定、可控的川芎药材,促进川芎产业的可持续发展。研究结果如下：
1.川芎苓种的调查、收集
在前期全国川芎种质资源调查的基础上,本文重点对四川省内川芎主要产区及苓种繁育地所涉及的5个市县20余个乡镇进行了实地调研,基本弄清了四川川芎苓种的分布和主产区的演迁、当地传统繁育技术、苓种保存方法、质量评判及产销情况等；调查表明,近年来,尤其是5.12汶川大地震后,发生了一些变迁,目前主要集中在都江堰市中兴镇、都江堰市青城后山红岩、都江堰市虹口乡,彭州市小渔洞镇、龙门山镇、通济镇,汶川县水磨镇、三江口镇,崇州市三郎镇,什邡市冰川镇等,海拔在900m-1500m。其中彭州小鱼洞镇的川芎苓种繁育面积较大。
主产区苓种的来源主要是坝区药农定点委托山区药农代为自繁自用,收获的苓种多数供坝区自我栽种,少数流入集市销售供其他栽种所需。川芎苓种市场上质量良莠不齐,无统一标准规范。另外,近年在一些川芎主产区和苓种集市上出现了少量“坝苓种”、“冻苓种”和“山区自栽苓种”,已在四川省内或调往省外作为川芎生产用种。
调查同时,收集了不同产地、不同集市的川芎苓种共34分,部分保留在前期建立的川芎种质资源圃中,部分提供给国家药用植物种质资源库,部分作为课题研究用。
2.川芎苓种贮藏及种质保存技术研究
针对川芎生产用苓种,首次进行了常温保存及低温贮藏技术研究,弄清了苓种在常温条件下存放6～10天,脱去的水分适中,不会影响发芽率,为川芎苓种采收后的短期贮藏提供了科学依据；建立了川芎苓种冷藏保存方法,可使川芎苓种短期保存时间延长至3个月,为该方法的进一步的推广应用,以满足川芎不同栽种地区对川芎苓种不同贮藏时间的需求及改变川芎苓种自繁自用,促使其商品化奠定了基础。
针对川芎种质中长期保存,首次采用限制生长保存法,以川芎茎段为组培材料,以MS为固体培养基,建立了川芎试管苗培养方法,对比了改变培养环境、调整培养基养分水平、添加渗透性化合物和生长抑制剂等实验,得到了川芎种质限制生长保存最佳方案。恢复生长实验表明保存6月后的种质能生长成活,保存苗和对照苗的形态特征与同工酶检测分析,没有发现变异现象,保持了良好的遗传稳定性,为实现川芎种质材料的中长期保存奠定了基础。
同时采用玻璃化法对川芎种质超低温保存的可行性进行探索,找到了适合的川芎玻璃化法的外植体是川芎茎尖,建立了川芎组培再生苗的培养方法,为进一步深入研究打下了基础。
3.川芎新品种选育研究
在前期川芎种质资源鉴定的基础上,构建成了由1300个株系,约8000个单株组成的育种用群体。利用该群体,采用系统选育的方法进行了川芎优良新品种选育,从优良单株选择、优良株系的筛选、品系比较试验、品种(系)多点试验,最终选育出川芎优良新品种“绿芎1号”,并通过四川省农作物品种审定委员会的药用植物新品种审定。该品种具有成株叶色浓绿,持绿期长,生长旺盛,群体整齐性、一致性较好,抗病性强的特点,并表现出良好的丰产性、稳定性和适应性,其药材质量符合《中国药典》的相关规定。为逐步实现川芎品种的标准化奠定了基础,为川芎药材的规范化生产提供了优良种源。
4.川芎苓种繁育技术研究
在前期研究的基础上,深入进行了川芎苓种的繁育技术规程的研究,包括适宜栽种区域、产地环境条件、选地与整地、抚芎的选择与处理、栽种、田间管理、病虫害防治、采收与采后处理等,确定了川芎苓种繁育各环节最适宜的方法和措施。在此基础上制订了川芎苓种繁育技术操作规程(SOP),为山区苓种繁育地优质苓种的生产提供了技术保障。
5.川芎苓种质量标准研究
首次进行了川芎苓种质量检验规程的研究,针对川芎繁殖材料的特殊性,结合产区药农传统经验,创新性的提出了以外观形态、百粒重、苓子系数、混杂率、病虫害等作为川芎苓种质量评价的综合指标,并制定了川芎苓种质量检验规程,内容包括真实性鉴定、形态指标、重量测定方法、苓子系数、混杂率、病虫害。为川芎苓种的质量评判提供了技术支撑。
首次进行了川芎苓种分级标准的研究,通过对比不同等级川芎苓种对其植株生长发育、药材产量和质量的影响,结果表明,不同等级苓种对川芎的生长有显著性的影响,并且在川芎的不同生长时期其影响因素有差异,一、二等苓种的川芎在植株生长发育及药材产量上均明显优于三等苓种的川芎；药材质量方面,不同等级苓种的川芎在药材性状、薄层色谱、主成分含量上无明显差异。在总结药农长期实践经验和前期研究的基础上,根据不同等级苓种对植株生长发育和药材产量、质量影响的实验研究结果,并结合生产实际,拟订了川芎苓种等级划分标准。
在上述研究基础上,起草了《川芎苓种质量标准(草案)》。为川芎苓种生产、流通、检验和使用提供了标准依据。
Chuanxiong Rhizoma, the rhizome of Ligusticum Chuanxiong Hort., is a famous trueborn Chinese medicinal materials in Sichuan, China which has been cultivating and using for two thousands years. Ligusticum Chuanxiong Hort. is an asexual reproductive plant. Its overground swollen node called Lingzhong is used as reproductive material in production. Because of lacking stain selection and breeding, its genetic characterization degenerated to some extent over thousands years of cultivation. In Sichuan, Chuanxiong Rhizoma's main production area, Lingzhong breeding is still absent of unified technology protocol and scientific organization and management. And the absence of classification criterion and testing rules for Lingzhong result in its uneven quality. Meanwhile, traditional natural conservation method is still widely used in production that can not preserve the medicinal herb for long. Traditional field conservation method consumes much manpower and material resources and may lead to the loss of germplasm, while relevant germplasm conservation technology is still lack of research.
Seed and seedling of good variety and good quality are warranties to realize good agricultural practice for Chinese medicinal materials which can ensure herb's quality in return. In view of the bottleneck in standardized production of Chuanxiong Rhizoma, the article took Lingzhong'standardization as the breakthrough point, studied the standardization of seedling of Ligusticum Chuanxiong Hort.(Lingzhong) and its germplasm conservation technology and aimed to promote Lingzhong's standardization, to provide Lingzhong of high quality for production of Chuanxiong Rhizoma, and finally to produce Chuanxiong Rhizoma with high, stable and controllable quality as well as high yield thus to promote the continuable development of Chuanxiong Rhizoma production industry. The research results are as follows:
1Lingzhong search and collecting
On the basis of early germplasm investigation around China, the article did field survey at Chuanxiong Rhizoma's main production and breeding area including more than20villages and towns belonging to5cities and counties of Sichuan. Its distribution, evolution and migration of main production area, local cultivation technology, germplasm breeding and conservation technology, quality evaluation method and marketing situation of Lingzhong in Sichuan are basically made clear. The search showed that its production area had alternated at resent years especially after Wenchuan earthquake. Lingzhong in Sichuan are mainly gathered at Zhongxing town, Hongyan at back peak of Qingcheng mountain, Hongkou village of Dujiangyan city, Xiaoyudong town, Longmenshan town, Tongji town of Pengzhou city, Shuimo town, Sanjiangkou town of Wenchan county, Sanlang town of Pengzhou city, Bingchuan town of shifang city, whose average elevation are between900m and1500m. Among them, Lingzhong's breeding area at Xiaoyudong town of Pengzhou city is bigger.
In Chuanxiong Rhizoma's major production area, Lingzhong is mainly bred by famers in mountainous area who are spot authorized by farmers in plain area. Most collected Lingzhong are planted at plain while few flew into the market for else need. Good and bad Lingzhong at the market are intermingled without unified criterion. Recent years, it was discovered that a few Ba Lingzhong, Dong Lingzhong, Moutain Lingzhong for own need in Chuanxiong Rhizoma's main production area as well as the market had flown into many planting area of Chuanxiong Rhizoma in and out of Sichuan province.
Lingzhong germplasm coming from34production areas and markets are colleted in the investigation which are partly preserved in early-established germplasm bank, partly provided to National Germplam Bank and partly stayed behind for project research.
2Lingzhong storage and germplasm conservation technology
Studied normal temperature storage technology and low temperature storage technology to conserve Lingzhong used for produciton. Results showed that Lingzhong could be conserved for6to10days under normal temperature which will not influence its germination percentage, which provided scientific references for its short term conservation after recovered. Established refrigeration conservation method that could prolong Lingzhong's short term conservation time to3months. This method lay a solid foundation for its further spreading and commercialization which can satisfy the need of different area's farmers for Lingzhong's different storage time.
Growth restriction conservation method was first applied to Chuanxiong tube seedling for its middle term conservation. Taking its stem as explant and MS as solid medium, Chuanxiong tube seedling's culture method was established. The article studied the influences on Chuangxiong tube seedlings by altering culture circumstance, adjusting medium nutrient composition, adding permeable compound and growth inhibitor. And the best growth restriction conservation method had been established. The recovering growth experiment showed that Chuanxiong germplasm could survive after six months'reservation. Variation was not found between preserved seedling and controlled seedling by morphological characteristics comparison and isoenzyme analysis. Conserved seedling had kept good heredity stability. This had been lay a good foundation for Lingzhong's middle term conservation.
Explored the practicability of preserving Lingzhong under ultralow temperature adopting embedding vitrification method. The best explant for embedding vitrification method is the stem tip. This has been established a culture method for Chuanxiong tissue culture seedling and lay a foundation for its further study.
3New variety breeding
Chuanxiong breed colony from1300fine individuals and8000single-plants is constructed on the basis of early Chuanxiong resouce identification. Using this colony, systematic selection method is adopted to breed new Chuanxiong variety of high quality. Over6six years, after good single-plant selection, good stains selection, strain comparison test and variety(stain) muti-location trials, Ivxiong1was finally selected out and certified by the Crops Variety Appraisal Committee of Sichuan Province.
Lvxiong I's mature plant grows dark green leaves that could be kept for a long time. It showed good colony regularity, strong disease-resistance, fine yielding ability, stability and adaptability. Quality of Chuanxiong Rhizoma from the strain fits to pharmacopeia's need. It had lain a foundation for Chuanxiong variety's standardization and provided good provenance for Chuanxiong Rizoma's standardized production.
4Lingzhong breeding technology
On the basis of early research, Lingzhong breeding technical specification including suitable plant area, environmental condition of producing area, field selection and soil preparation, Fuxiong's selection and treatment, planting, field management, extermination of disease and insect pest, processing in production place and so on were deeply researched. The best methods and measures in every process of Lingzhong breeding were determined. Standard operation practicee for Lingzhong breeding is formulated on basis of it, and this provide technical support for the production of fine Lingzhong at mountain breeding area.
5Quality standard study of Lingzhong
Studied the quality testing protocol of Lingzhong. Taking account into Chuanxiong Rhizoma breeding material's particularity and using medicinal herb grower' traditional experience for references, comprehensive indexes were put forward to evaluate Lingzhong's quality which include morphological appearance,100grain weight, seedling coefficient, mixture rate, insect pest and disease. And quality testing protocol of Lingzhong was formulated which contains authenticity identification, shape index, weight determination method, seedling coefficient, impurity rate and insect pest and disease. It provide technical support for the evaluation of quality of Lingzhong.
First studied classification criterion of Lingzhong by compare influences of Lingzhong of different classification on its growth, yield and quality. The result showed that classification significantly influenced its growth and the influence factor varies at its different growth period. The first and second classification apparently excel third classification at Chuanxiong's growth and yield. Different classification showed little differences at the herbal properties, Thin-Layer Chromatography, content of main component. Through summarizing herb farmer's long practical experience and early research achievement, and referring to the research result above as well as actual production, Lingzhong classification criterion was drawn up.
On the basis of above research results, Chuanxiong Lingzhong Quality Criterion (draft) was drawn up. It provide standard criterion for Chuanxiong Lingzhong's production, circulation, examination and application.
中药材质量稳定需要中药材生产的规范化,中药材生产规范化需要中药材种子种苗的标准化。优良品种和优质的种子种苗是实现中药材规范化生产的基础和源头。本文针对川芎规范化生产中的薄弱环节,以苓种标准化问题为切入点,开展了川芎苓种的标准化研究和种质保存技术研究,以推动川芎苓种的标准化进程,为川芎规范化生产提供源头保障。以生产出优质、高产、稳定、可控的川芎药材,促进川芎产业的可持续发展。研究结果如下：
1.川芎苓种的调查、收集
在前期全国川芎种质资源调查的基础上,本文重点对四川省内川芎主要产区及苓种繁育地所涉及的5个市县20余个乡镇进行了实地调研,基本弄清了四川川芎苓种的分布和主产区的演迁、当地传统繁育技术、苓种保存方法、质量评判及产销情况等；调查表明,近年来,尤其是5.12汶川大地震后,发生了一些变迁,目前主要集中在都江堰市中兴镇、都江堰市青城后山红岩、都江堰市虹口乡,彭州市小渔洞镇、龙门山镇、通济镇,汶川县水磨镇、三江口镇,崇州市三郎镇,什邡市冰川镇等,海拔在900m-1500m。其中彭州小鱼洞镇的川芎苓种繁育面积较大。
主产区苓种的来源主要是坝区药农定点委托山区药农代为自繁自用,收获的苓种多数供坝区自我栽种,少数流入集市销售供其他栽种所需。川芎苓种市场上质量良莠不齐,无统一标准规范。另外,近年在一些川芎主产区和苓种集市上出现了少量“坝苓种”、“冻苓种”和“山区自栽苓种”,已在四川省内或调往省外作为川芎生产用种。
调查同时,收集了不同产地、不同集市的川芎苓种共34分,部分保留在前期建立的川芎种质资源圃中,部分提供给国家药用植物种质资源库,部分作为课题研究用。
2.川芎苓种贮藏及种质保存技术研究
针对川芎生产用苓种,首次进行了常温保存及低温贮藏技术研究,弄清了苓种在常温条件下存放6～10天,脱去的水分适中,不会影响发芽率,为川芎苓种采收后的短期贮藏提供了科学依据；建立了川芎苓种冷藏保存方法,可使川芎苓种短期保存时间延长至3个月,为该方法的进一步的推广应用,以满足川芎不同栽种地区对川芎苓种不同贮藏时间的需求及改变川芎苓种自繁自用,促使其商品化奠定了基础。
针对川芎种质中长期保存,首次采用限制生长保存法,以川芎茎段为组培材料,以MS为固体培养基,建立了川芎试管苗培养方法,对比了改变培养环境、调整培养基养分水平、添加渗透性化合物和生长抑制剂等实验,得到了川芎种质限制生长保存最佳方案。恢复生长实验表明保存6月后的种质能生长成活,保存苗和对照苗的形态特征与同工酶检测分析,没有发现变异现象,保持了良好的遗传稳定性,为实现川芎种质材料的中长期保存奠定了基础。
同时采用玻璃化法对川芎种质超低温保存的可行性进行探索,找到了适合的川芎玻璃化法的外植体是川芎茎尖,建立了川芎组培再生苗的培养方法,为进一步深入研究打下了基础。
3.川芎新品种选育研究
在前期川芎种质资源鉴定的基础上,构建成了由1300个株系,约8000个单株组成的育种用群体。利用该群体,采用系统选育的方法进行了川芎优良新品种选育,从优良单株选择、优良株系的筛选、品系比较试验、品种(系)多点试验,最终选育出川芎优良新品种“绿芎1号”,并通过四川省农作物品种审定委员会的药用植物新品种审定。该品种具有成株叶色浓绿,持绿期长,生长旺盛,群体整齐性、一致性较好,抗病性强的特点,并表现出良好的丰产性、稳定性和适应性,其药材质量符合《中国药典》的相关规定。为逐步实现川芎品种的标准化奠定了基础,为川芎药材的规范化生产提供了优良种源。
4.川芎苓种繁育技术研究
在前期研究的基础上,深入进行了川芎苓种的繁育技术规程的研究,包括适宜栽种区域、产地环境条件、选地与整地、抚芎的选择与处理、栽种、田间管理、病虫害防治、采收与采后处理等,确定了川芎苓种繁育各环节最适宜的方法和措施。在此基础上制订了川芎苓种繁育技术操作规程(SOP),为山区苓种繁育地优质苓种的生产提供了技术保障。
5.川芎苓种质量标准研究
首次进行了川芎苓种质量检验规程的研究,针对川芎繁殖材料的特殊性,结合产区药农传统经验,创新性的提出了以外观形态、百粒重、苓子系数、混杂率、病虫害等作为川芎苓种质量评价的综合指标,并制定了川芎苓种质量检验规程,内容包括真实性鉴定、形态指标、重量测定方法、苓子系数、混杂率、病虫害。为川芎苓种的质量评判提供了技术支撑。
首次进行了川芎苓种分级标准的研究,通过对比不同等级川芎苓种对其植株生长发育、药材产量和质量的影响,结果表明,不同等级苓种对川芎的生长有显著性的影响,并且在川芎的不同生长时期其影响因素有差异,一、二等苓种的川芎在植株生长发育及药材产量上均明显优于三等苓种的川芎；药材质量方面,不同等级苓种的川芎在药材性状、薄层色谱、主成分含量上无明显差异。在总结药农长期实践经验和前期研究的基础上,根据不同等级苓种对植株生长发育和药材产量、质量影响的实验研究结果,并结合生产实际,拟订了川芎苓种等级划分标准。
在上述研究基础上,起草了《川芎苓种质量标准(草案)》。为川芎苓种生产、流通、检验和使用提供了标准依据。
Chuanxiong Rhizoma, the rhizome of Ligusticum Chuanxiong Hort., is a famous trueborn Chinese medicinal materials in Sichuan, China which has been cultivating and using for two thousands years. Ligusticum Chuanxiong Hort. is an asexual reproductive plant. Its overground swollen node called Lingzhong is used as reproductive material in production. Because of lacking stain selection and breeding, its genetic characterization degenerated to some extent over thousands years of cultivation. In Sichuan, Chuanxiong Rhizoma's main production area, Lingzhong breeding is still absent of unified technology protocol and scientific organization and management. And the absence of classification criterion and testing rules for Lingzhong result in its uneven quality. Meanwhile, traditional natural conservation method is still widely used in production that can not preserve the medicinal herb for long. Traditional field conservation method consumes much manpower and material resources and may lead to the loss of germplasm, while relevant germplasm conservation technology is still lack of research.
Seed and seedling of good variety and good quality are warranties to realize good agricultural practice for Chinese medicinal materials which can ensure herb's quality in return. In view of the bottleneck in standardized production of Chuanxiong Rhizoma, the article took Lingzhong'standardization as the breakthrough point, studied the standardization of seedling of Ligusticum Chuanxiong Hort.(Lingzhong) and its germplasm conservation technology and aimed to promote Lingzhong's standardization, to provide Lingzhong of high quality for production of Chuanxiong Rhizoma, and finally to produce Chuanxiong Rhizoma with high, stable and controllable quality as well as high yield thus to promote the continuable development of Chuanxiong Rhizoma production industry. The research results are as follows:
1Lingzhong search and collecting
On the basis of early germplasm investigation around China, the article did field survey at Chuanxiong Rhizoma's main production and breeding area including more than20villages and towns belonging to5cities and counties of Sichuan. Its distribution, evolution and migration of main production area, local cultivation technology, germplasm breeding and conservation technology, quality evaluation method and marketing situation of Lingzhong in Sichuan are basically made clear. The search showed that its production area had alternated at resent years especially after Wenchuan earthquake. Lingzhong in Sichuan are mainly gathered at Zhongxing town, Hongyan at back peak of Qingcheng mountain, Hongkou village of Dujiangyan city, Xiaoyudong town, Longmenshan town, Tongji town of Pengzhou city, Shuimo town, Sanjiangkou town of Wenchan county, Sanlang town of Pengzhou city, Bingchuan town of shifang city, whose average elevation are between900m and1500m. Among them, Lingzhong's breeding area at Xiaoyudong town of Pengzhou city is bigger.
In Chuanxiong Rhizoma's major production area, Lingzhong is mainly bred by famers in mountainous area who are spot authorized by farmers in plain area. Most collected Lingzhong are planted at plain while few flew into the market for else need. Good and bad Lingzhong at the market are intermingled without unified criterion. Recent years, it was discovered that a few Ba Lingzhong, Dong Lingzhong, Moutain Lingzhong for own need in Chuanxiong Rhizoma's main production area as well as the market had flown into many planting area of Chuanxiong Rhizoma in and out of Sichuan province.
Lingzhong germplasm coming from34production areas and markets are colleted in the investigation which are partly preserved in early-established germplasm bank, partly provided to National Germplam Bank and partly stayed behind for project research.
2Lingzhong storage and germplasm conservation technology
Studied normal temperature storage technology and low temperature storage technology to conserve Lingzhong used for produciton. Results showed that Lingzhong could be conserved for6to10days under normal temperature which will not influence its germination percentage, which provided scientific references for its short term conservation after recovered. Established refrigeration conservation method that could prolong Lingzhong's short term conservation time to3months. This method lay a solid foundation for its further spreading and commercialization which can satisfy the need of different area's farmers for Lingzhong's different storage time.
Growth restriction conservation method was first applied to Chuanxiong tube seedling for its middle term conservation. Taking its stem as explant and MS as solid medium, Chuanxiong tube seedling's culture method was established. The article studied the influences on Chuangxiong tube seedlings by altering culture circumstance, adjusting medium nutrient composition, adding permeable compound and growth inhibitor. And the best growth restriction conservation method had been established. The recovering growth experiment showed that Chuanxiong germplasm could survive after six months'reservation. Variation was not found between preserved seedling and controlled seedling by morphological characteristics comparison and isoenzyme analysis. Conserved seedling had kept good heredity stability. This had been lay a good foundation for Lingzhong's middle term conservation.
Explored the practicability of preserving Lingzhong under ultralow temperature adopting embedding vitrification method. The best explant for embedding vitrification method is the stem tip. This has been established a culture method for Chuanxiong tissue culture seedling and lay a foundation for its further study.
3New variety breeding
Chuanxiong breed colony from1300fine individuals and8000single-plants is constructed on the basis of early Chuanxiong resouce identification. Using this colony, systematic selection method is adopted to breed new Chuanxiong variety of high quality. Over6six years, after good single-plant selection, good stains selection, strain comparison test and variety(stain) muti-location trials, Ivxiong1was finally selected out and certified by the Crops Variety Appraisal Committee of Sichuan Province.
Lvxiong I's mature plant grows dark green leaves that could be kept for a long time. It showed good colony regularity, strong disease-resistance, fine yielding ability, stability and adaptability. Quality of Chuanxiong Rhizoma from the strain fits to pharmacopeia's need. It had lain a foundation for Chuanxiong variety's standardization and provided good provenance for Chuanxiong Rizoma's standardized production.
4Lingzhong breeding technology
On the basis of early research, Lingzhong breeding technical specification including suitable plant area, environmental condition of producing area, field selection and soil preparation, Fuxiong's selection and treatment, planting, field management, extermination of disease and insect pest, processing in production place and so on were deeply researched. The best methods and measures in every process of Lingzhong breeding were determined. Standard operation practicee for Lingzhong breeding is formulated on basis of it, and this provide technical support for the production of fine Lingzhong at mountain breeding area.
5Quality standard study of Lingzhong
Studied the quality testing protocol of Lingzhong. Taking account into Chuanxiong Rhizoma breeding material's particularity and using medicinal herb grower' traditional experience for references, comprehensive indexes were put forward to evaluate Lingzhong's quality which include morphological appearance,100grain weight, seedling coefficient, mixture rate, insect pest and disease. And quality testing protocol of Lingzhong was formulated which contains authenticity identification, shape index, weight determination method, seedling coefficient, impurity rate and insect pest and disease. It provide technical support for the evaluation of quality of Lingzhong.
First studied classification criterion of Lingzhong by compare influences of Lingzhong of different classification on its growth, yield and quality. The result showed that classification significantly influenced its growth and the influence factor varies at its different growth period. The first and second classification apparently excel third classification at Chuanxiong's growth and yield. Different classification showed little differences at the herbal properties, Thin-Layer Chromatography, content of main component. Through summarizing herb farmer's long practical experience and early research achievement, and referring to the research result above as well as actual production, Lingzhong classification criterion was drawn up.
On the basis of above research results, Chuanxiong Lingzhong Quality Criterion (draft) was drawn up. It provide standard criterion for Chuanxiong Lingzhong's production, circulation, examination and application.
引文
[1]魏建和,陈士林,程惠珍,等.中药材种子种苗标准化工程[J].世界科学技术-中医药现代化,2005,7(6)：104-108
[2]温学森,魏建和,杨世林,等.地黄种质资源的离体保存研究[J].中国中药杂志,2003,28(1)：17-20
[3]国家药典委员会.中华人民共和国药典(2010年版)一部[S].北京：化学工业出版社,2010：38
[4]贾敏如.川芎、川白芷生产质量管理规范(GAP)的研究[M].成都：四川科学技术出版社,2007
[5]王昌华,刘翔,银福军,等.大黄种子质量分级标准的研究[J].时珍国医国药2009,20(7)：1605-1606
[6]孙群,杨力钢,丁自勉,等.乌拉尔甘草种子质量分级标准的研究[J].中国中药杂志,2008,33(10)：1126-1129
[7]贺玉林,李先恩,淡红梅.远志种子质量分级标准研究[J].种子,2007,26(1)：106-107
[8]贺玉林,李先恩,淡红梅,等.远志种子检验规程[J].中国中药杂志,2007,32(15)：1497-1500
[9]吾拉尔古丽,王建华,李先恩.板蓝根种子发芽试验标准化研究[J].种子,2005,24(6)：34-36
[10]胡榜文,薛江涛,李国峰,等.玄参脱毒种苗生产标准操作规程[J].现代中药研究与实践,2008,22(6)：17-19
[11]陈翠,康平德,杨丽云,等.云南重楼种苗繁育技术[J].中国现代中药,2010,12(2)：23-24,28
[12]史永忠,潘瑞炽,王小菁,等.铁皮石斛种质室温离体保存[J].华南师范大学学报,1999,(4)：73-77
[13]黄菊辉,陈世儒.生姜种质资源离体保存[J].西南农业大学学报,1996,13(3)：310-312
[14]李海兵,周娜,赵姣,等.怀山药种质资源的包埋玻璃化超低温保存与植株再生[J].植物学报,2010.45(3)：379-383
[15]孔令婕,巢建国,谷巍,等.茅苍术种质资源的超低温保存[J].南京中医药大学学报,2010,26(1)：56-58
[16]陈辉,陈晓玲,陈龙清,等.百合种质资源限制生长法保存研究[J].园艺学报,2006,33(4)：789-793
[17]雷志强,张寿文,刘华,等.车前种子种苗的分级标准研究[J].江西中医学院学报,2007,19(5)：65-67
[18]张春庆,王建华.种子检验学[M].北京：高等教育出版社,2005
[19]陈火英.现代种子种苗学[M].上海：上海科学技术出版社,1999
[20]洪森荣,郭连金.离体保存技术在植物种质资源保存中的应用[J].上饶师范学院学报,2006,26(3)：92-95
[21]张巧仙,范冬梅.植物种质资源的保存——离体保存.太原师范学院学报(自然科学版[J].2005,4(3)：82-85
[22]陈晓明,刘海龙,蔡玲.桉树种质资源离体保存技术[J].广西林业科学,2009,38(1)：8-13
[23]胡炜,孙拥军.强化对种子质量及指标体系的认识[J].种子科技,2003,(4)：213
[24]黄勇.石斛属植物种质资源离体保存.湖北农业科技[J].2009,48(11)：2716-2718
[25]黄璐琦.分子生药学[M].北京：北京医科大学出版社,2000
[26]Jarret R L,Florlowski W J. In Vitro Active Vs.Field Genbank Maintenance of Sweet Potato Germplasm:Major Costs and Considerrations [J]. Hort Science, 1990,25(2):141
[27]董玉琛.保护作物多样性,发掘有用新基因——试论我国作物种质资源研究当前的任务[J].植物遗传资源科学,2000.1(1)：2
[28]马小军,肖培根.种质资源遗传多样性在药用植物开发中的重要意义[J].中国中药杂志,1998,23(10)：579
[29]周逊,向长萍.植物种质资源缓慢生长离体保存研究进展[J].中国蔬菜,2008,(11)39-41
[30]伊华林,邓秀清.植物种质离体保存技术研究进展[J].植物学通报,1999,16(5)：574-581
[31]付传明,赵志国,黄宁珍,等.药用植物红根草种质资源的离体保存[J].广西植物,2007,27(4)：653-657
[32]刘小莉,杨耀文,钱子刚.多效唑和蔗糖对金铁锁离体保存的影响[J].云南中医学院学报,2009,34-36
[33]李明军,徐鑫,夏民,等.PP333和BA组合对怀地黄试管苗生长发育的影响[J].植物学通报,2006,23(1)：56-59
[34]李峰,付传明,黄宁珍,等.巴戟天种质离体保存研究[J].广西植物,2008,28(1)：95-99
[35]Wang JF,Liu YX,Lin SQ.A study of the preservation in vitro of loquat germplasm Ⅱ:Effect of plant growth inhibitor[J].Subtrop Plant Sci,2002,31(4):1-4
[36]Kartha K K,Mroginski L A,Pahl K K,et al.Gernplassm preservation of coffea (Coffea arabica)by in vitro curure of shoot apical meristems[J].Plant Science Letters,1981,22:411-413
[37]Bertrand Desbnutais A,Noirot M,Charrier A.Slow growth invitro conservation of coffee (Coffea spp) [J]. Plant Cell,Tissue and Organ Culture,1992,31:105-110
[38]张玉进,张兴国,刘佩英.魔芋不定芽的低温保存研究[J].西南农业大学学报,1999,21(4)：303-306
[39]张玉进,张兴国,刘佩英.魔芋花粉的低温和超低温保存[J].园艺学报,2000,27(2)：139-140
[40]徐刚标,何方,陈良昌.银杏种质离体保存的研究[J].经济林研究,1999,17(4)：15-18
[41]徐刚标,何方,黄晓光.银杏种质离体保存的研究Ⅰ.银杏花粉贮存[J].中南林学院学报,2000,20(1)：27-30
[42]刘剑锋,阎秀峰,程云清,等.高山红景天(Rhodiola sachalinensis)试管苗缓慢生长法保存及试管苗DNA含量分析[J].浙江大学学报(农业与生命科学版),2007,33(4)：373-378
[44]刘小莉,杨耀文,钱子刚.多效唑和蔗糖对金铁锁离体保存的影响[J].云 南中医学院学报,2009,32(1)：34-36
[45]李锋,付传明,黄宁珍,等.巴戟天种质离体保存研究[J].广西植物,2008,28(1)：95-99
[46]付传明,黄宁珍,赵志国,等.广西地不容种质离体保存技术研究[J].广西科学,2007,14(2)：155-159
[47]史永忠,潘瑞炽,王小菁,等.铁皮石斛种质室温离体保存[J].华南师范大学学报(自然科学版),1999,(4)：73-77
[48]张晓丽,刘文英,张楠,等.PP333对怀地黄种质离体保存的影响[J].河南师范大学学报(自然科学版),2009,(3)：171-174
[49]唐凤鸾,韦记青,蒋运生,等.黄花蒿组培快繁与种质离体保存的研究[J].热带亚热带植物学报,2008,16(5)：486-490
[50]付传明,黄宁珍,唐凤鸾,等.山银花的组培快繁和种质离体保存研究[J].广西科学,2008,15(3)：304-308
[51]洪森荣,尹明华,柯维忠,等.PP333对野葛离体保存的影响[J].上饶师范学院学报,2007,27(2)：68-72
[52]张玉进,张若国,庞杰,等.魔芋茎尖玻璃化冻存研究[J].作物学报,2001,27(1)：97-102
[53]徐刚标,易文,何方,等.银杏种质离体保存的研究：Ⅱ.银杏胚超低温保存[J].中南林学院学报,2000,20(2)：7-10
[54]曾华兰,叶鹏盛,倪国成,等.川芎主要病虫害及其发生危害规律研究[J].西南农业学报,2009,22(1)：99-101
[55]张萃蓉,杨星勇,张玉芳,等.川芎块茎腐烂病研究[J].云南农业大学学报,1993,8(3)：259-261
[56]张俊,蒋桂华,敬小莉,等.我国药用植物种质资源离体保存研究进展[J].世界科学技术,2011,12(3)23
[57]张俊,蒋桂华,敬小莉,等.我国药用植物育种的最新进展[J].中国现代中药,2010,12(5)：5-9
[58]高农,韩学俭.川芎苓子繁育技术[J].特种经济动植物,2004,10：26
[59]王岚,肖海波,马逾英.川芎道地性的ISSR分析[J].四川大学学报(自 然科学版),2008,45(6)：1472-1476
[60]张丽萍,杨世林,杨春清,等.我国药材种子种苗产业存在的问题及其对策[J].中国中药杂志,1999,24(10)：579-581
[61]张廷红,郭永杰,当归种苗繁殖技术[J].甘肃农业科技,1997,8：14-15
[62]崔秀明,王朝梁,陈中坚.种苗分级对三七生长和产量的影响[J].中药材,1998,21(2)：60-61
[63]朱贞丽,王鼎峰,李勇,等.马蓝优质种苗培养技术的研究[J].海峡药学,2009,21(4)：87-88
[64]田伟,温春秀,周巧梅,等.不同来源远志种子的质量比较[J].现代中药研究与实践,2006,20(5)：19-20
[65]第九届全国人民代表大会常务委员会第十六次会议通过.中华人民共和国种子法.2000,7
[66]洪森荣,李明军.玻璃化法超低温保存怀山药种质的技术研究[J].中草药,2006,37(11)：1715-1718
[67]刘剑锋,程云清,秦小伟.高山红景天茎尖的包埋-玻璃化超低温保存研究[J].,吉林师范大学学报(自然科学版),2009,3：61-65
[68]Leunufina S,Keller E R J.Investigating a new cryopreservation protocol for yams (Diooscorea spp.) [J]. Plant Cell Reports,2003,21 (12):1159-1166
[69]Shibili R A,Moges A D,Karam N S.Cryopreservation of Africa violet (Sain tpaulia Ionantha wendl.) shoot tips [J]. In Vitro Cell Dev Biol Plant,2004,40(4):389-395
[70]Hirai D,Sakai A,Cryopreservation of shoot tips from in vitro plants of sweet potata[Ipomoea botatas (L.)Lam.] by vitrification [J]. Plant Cell Reports,2003,19(7):733-737
[71]Liu Y G, Wang X Y.Study of cyropreservation technique of apple germplasm by vitrification [J].J Shandong Agric Univ:Nat Sci, 2002,33(1):32-36
[72]Matsumoto T,Sakai A,Takahashi C.Cryopreservation of in vitro-grown spical meristems of wasabi (Wasabia japonica) by encapsulation-vitrifacation method[J].Cryo-Letters,1995,16(4):189-196
[73]Matsumoto T,Sakai A,Yamada K.Cryopreservation of in vitro-grown spical meristems of wasabi (Wasabia japonica) by vitrifacation and subsequent high plant regeneration [J]. Plant Cells Reports,1994,13 (8):442-446
[74]Gupta S, Reed B M.Cryopreservation pf shoot tips of blackberry and raspberry by encapsulation-dehudtation and vitrification [J]. Cryo-letters,2006, 27(1):29-42
[75]张志珂,陈彬,杨凤玲,等.枇杷野生种的种质资源离体保存初探[J].复件果树,2008,3：28-32
[76]刘小军.枇杷种质离体保存的初步研究[D].福建：福建农林大学,2002
[77]Zheng S Q.Achievement and Prospect of Loquat Breeding in China[J].Acta Horticulturae,2007,705:85-92
[78]吴卫,郑有良,陈黎,等.川产鱼腥草种质资源的同工酶分析.中草药,2002,25(10)：695-698
[79]王永炎.有关中药材GAP中种质资源的若干问题剖析.第四届北京国际周论坛报告集
[80]中国药材GAP网,首页>GAP研究与实践>药用植物栽培技术.药用植物栽培(四)--药用植物的良种选育
[81]陈兴福,丁德蓉,黄文秀,等.川芎生长发育特性的研究[J].中国中药杂志,1997,22(9)：527-529
[82]黄璐琦,郭兰萍,崔光红,等.中药资源可持续利用的基础理论研究[J].中药研究与信息,2005,7(8)：4-7
[83]蒋桂华,马逾英,侯嘉,等.川芎种质资源的调查收集与保存研究[J].中草药,2008,39(4)：601-604
[84]郭才,霍志军.植物遗传育种及种苗繁育[M].北京：中国农业大学出版社,2006
[85]胡延吉.植物育种学[M].北京：高等教育出版社,2003
[86]张春庆,王建华.种子检验学[M].北京：高等教育出版社,2005
[87]喻方圆,周景莉.林木种苗质量检验技术[M].北京：中国林业出版社,2008
[1]贾敏如.川芎、川白芷生产质量管理规范(GAP)的研究[M].成都：四川科学技术出版社,2007：1
[2]万德光,彭成,赵军宁.四川道地中药材志[M].成都：四川科学技术出版社,2005：4
[3]苏颂.图经本草(胡乃长,王致谱辑注)[M].福建：福建科学技术出版社,1987：129
[4]刘圆,贾敏如.川芎品种、产地的历史考证[J].中药材.2001：24(5)：365
[5]彭县县志办公室.彭县县志(1910-1986)[M].成都：四川人民出版社,1986
[6]郭平,李祖伦,陈红,等.川芎地上部分挥发油化学成分的研究[J].中国中药杂志,1993,18(9)：551-552
[7]任东春,杨念云,钱士辉,等.川芎地上部分化学成分研究[J].中国中药杂志,2007,32(14)：1418-1420
[8]刘友平,李祖伦,张廷模,等.薄层扫描法测定川芎不同部位阿魏酸的含量[J].中国中药杂志,1995,20(1)：9-10
[9]徐杰军.常用中药药理与临床[M].成都：四川科学技术出版社.2005：25-26
[10]凌婧.川芎挥发油对大鼠脑缺血再灌注损伤的保护作用及其机制研究,四川大学硕士论.文,2006年(2003级)
[11]胡荣.川芎挥发油解热作用及机理研究,成都中医药大学硕士论文,2001年4月
[12]蒋桂华,马逾英,侯嘉,等.川芎种质资源的调查收集与保存研究[J].中草药,2008,39(4)：601-604
[13]王岚,肖海波,马逾英,等.川芎道地性的ISSR分析[J].四川大学学报(自然科学版),2008,45(6)：1472-1476
[14]赵勇,傅体华,范巧佳.川芎各器官的生长动态分析[J].西南农业学报.2008,21(4)：
[15]张毅,范巧佳,郑顺林,等.春季追肥对川芎产量、阿魏酸和总生物碱含量的影响[J].中国中医药杂志.2008,33(16)：1944-1947
[16]冯茜,黄云,巩春梅,等.川芎根腐病菌(Fusarium solani)的生物学特性[J].四川农业大学学报.2008,26(1)：24-27
[17]冯茜,何苗,黄云,等.川芎根腐病的症状及病原鉴定[J].植物保护学报.2008,35(4)：377-378
[18]陈要臻,王岚,马逾英,等.川芎简单序列重复间区多态性和扩增片段长度多态性分子 标记技术多态性比较研究[J].时珍国医国药,2009,20(6)：1422-1424
[19]马玎,马逾英,张利,等.三个不同产地的川芎与其近缘植物藁本的挥发油成分对比分析[J].中国现代中药.2009,11(7)：20-22,25
[20]王妙闻,张艺,张静,等.HPLC测定川芎中的总阿魏酸[J].华西药学杂志,2008,23(1)：100-102
[21]侯嘉,马逾英,贾敏如,等.川产道地药材川芎与云南、甘肃产川芎形态组织学研究[J].湖南中医药大学学报,2007,27(S1)：9
[22]蒋卫东,唐琳,陈鹏等. 川芎的组织培养研究. [J]安徽农业科学,2007,35(27)：8448-8450
[23]蒋卫东,唐琳,杨苍劲,等.卡拉霉素对川芎愈伤组织诱导和生长的影响[J].河南农业科学,2007(11)：85-87
[24]詹玉莲,马小军,戴均贵,等.川芎愈伤组织及悬浮培养物化学成分气相色谱-质谱联用分析[J].时珍国医国药, 2006,17(6)：1117-1119
[25]刘玉萍,曹晖,韩桂茹,等.中日产川芎的matK. ITS基因序列及其物种间的亲缘关系[J].药学学报,2002,37(1)：63-68
[26]魏建和,陈士林,程惠珍,等.中药材种子种苗标准化工程[J].世界科学技术-中医药现代化,2005,7(6)：104-108
[27]崔秀明,王朝梁,陈中坚.种苗分级对三七生长和产量的影响[J].中药材,1998,21(2)：60-61
[28]张国荣,张玉进,李生彬.乌拉尔甘草种子种苗分级标准制定[J].现代中药研究与实践,2004,18(4)：14-16
[29]张廷红,郭永杰,当归种苗繁殖技术[J].甘肃农业科技,1997,8：14-15
[30]朱贞丽,王鼎峰,李勇,等.马蓝优质种苗培养技术的研究[J].海峡药学,2009,21(4)：87-88
[31]Lyndsey A, Withers A. In vitro approaches to conversation of plant genetic resources [A]. In:Plant Tissue Culture and Its Agricultural Applications (LyndscyA. Withers and P. G. Aldrrsoneds) [C], London:Butterworths,1986:261-276
[32]Hawkes J G.Germplasm collection, preservation and use [A]. In:Plant Breeding[CKenneth J. Freg ed.]. [C]. New Delhi:Gayatri Offsite Press 1981:57-84
[33]史永忠,潘瑞炽,王小菁,等.铁皮石斛种质室温离体保存[J].华南师范大学学报(自然科学版),1999,(4)：73-77
[34]唐凤鸾,韦记青,蒋运生,等.黄花蒿组培快繁与种质离体保存的研究[J].热带亚热带植物学报,2008,16(5)：486-490
[35]李海兵,周娜,赵姣,等.怀山药种质资源的包埋玻璃化超低温保存与植株再生[J].植物学报,2010.45(3)：379-383
[36]孔令婕,巢建国,谷巍,等.茅苍术种质资源的超低温保存[J].南京中医药大学学报,2010,26(1)：56-58
[37]陈辉,陈晓玲,陈龙清,等.百合种质资源限制生长法保存研究[J].园艺学报,2006,33(4)：789-793
[38]洪森荣,郭连金.离体保存技术在植物种质资源保存中的应用[J].上饶师范学院学报,2006,26(3)：92-95
[39]Mantell S H, Smith H. Plant biotechnology[M]. Cambridge:Cambridge University Press,1983.187-218
[40]张玉进,张若国,庞杰,等.魔芋茎尖玻璃化冻存研究[J].作物学报,2001,27(1)：97-102
[41]徐刚标,何方,黄晓光.银杏种质离体保存的研究：Ⅰ.银杏花粉贮存[J].中南林学院学报,2000,20(1)：27-30
[42]徐刚标,易文,何方,等.银杏种质离体保存的研究：Ⅱ.银杏胚超低温保存[J].中南林学院学报,2000,20(2)：7-10
[43]高山林.药用植物遗传育种的现状与展望[J].世界科学技术：中药现代化,2001,3(6)：58-62
[44]徐昭玺,魏建和,冯秀娟,等.边条人参新品种的系统选育[J].中国医学科学院学报,2001,23(6)：542-546
[45]朱培林,吴永忠,兰冬生,等.江枳壳优良类型选择研究[J].现代中药研究与实践,2004,18(6)：24-26
[46]何先元,郭巧生,徐文斌,等.不同药用白菊花栽培品种田间试验研究[J].安徽农业大学学报,2005,32(3)：385-388
[47]王秋颖,郭顺星.天麻优良品种选育的初步研究[J].中国中药杂志,2001,26(11)：744-746
[48]王锦秀,赵健,黄占明.枸杞与番茄属间远缘杂交研究初报[J].宁夏农林科技,2005,(3)：8-9
[49]吴才祥,杨晟永,葛芝富.天麻远缘杂交育种初报[J].湖南林业科技,2007,34(1)：23-25
[50]唐宁.白术辐射诱变育种研究[J].安徽农学通报,2007,13(23)：84-85
[51]唐宁,张敦林.白术辐射诱变株系抗氧化酶活性研究[J].安徽农学通报,2007,13(24)：54-55
[52]唐宁.白术辐射诱变株系中白术内酯Ⅰ、Ⅲ的含量测定[J].安徽农学通报,2009,15(1)：45-45,19
[53]颉红梅,刘效瑞,李文建,等.甘肃当归新品系DGA2000-02的选育研究[J].原子核物理评论,2008,25(2)：196-200
[54]甄卫军,孙月华,马凤云.低能离子注入麻黄种子生物效应初步研究[J].种子,2003,(2)：26-27,44
[55]潘燕,肖翔,吴李君,等.N+离子注入对干旱胁迫条件下甘草幼苗的SOD酶和CAT酶活性及丙二醛含量的影响[J].激光生物学报,2005,14(6)：442--446
[56]魏胜林,吴李君,谢传晓,等.N+注入对甘草叶片腺体和腺体分泌多糖及叶片多糖的影响[J].核技术,2008,25(2)：196-200
[57]李耀维,武增寿.激光诱变选育雷公藤次生物质高产细胞系[J].激光生物学报,2000,9(4)：281-284
[58]郭斌,尉亚辉,等He-Ne激光诱变选育高产白藜芦醇细胞系[J].光子学报,2002,31(3)：277-280
[59]单成钢,王志芬,苏学合,等.航天诱变黄芩种子对其SP1代的影响[J].核农学报,2008,22(2)：188-191
[60]王志芬,苏学合,闫树林,等.太空搭载桔梗种子SP1代的生物学效应研究[J].核农学报,2004,18(4)：323-324
[61]王志芬,单成钢,苏学合,等.丹参种子航天搭载的诱变效应研究[J].现代中药研究与实践,2007,21(4)：6-8
[62]李金贵,朱奎,沈海玉,等.卫星搭载牛膝种子SP1植株的生物学特性[J].中国农学通报,2007,23(7)：42-45
[63]熊大胜,朱金桃,张自亮,等.三叶木通理化诱变技术及其成熟期变异研究[J].常德师范学院学报(自然科学版),2001,13(4)：79-80,89
[64]张秀省,张荣涛,曹岚,等.EMS诱变的长春花细胞系突变研究[J].中草药,2004,25(11)：1293-1296
[65]孔青,刘林,李华斌.药用植物女贞的快繁[J].山东林业科技,2008,38(3)：69-70
[66]瞿大枞,陈崇顺,王轶,等.药用植物半枝莲的离体快繁[J].安徽农业科学,2007,35 (17)：5065-5066,5095
[67]唐伟斌,石晓云.药用植物桔梗组培快繁体系的建立[J].河北师范大学学报(自然科学版),2006,30(5)：599-601
[68]刘建成,陈先玉.川西獐牙菜的组织培养与快速繁殖[J].植物生理学通讯,2003,39(3)： 237-237
[69]温春秀,谢晓亮,吴志明,等.丹参脱病毒及组培快繁技术研究[J].中草药,2004,35(9)：1057-1059
[70]张寿文,雷志强,刘华,等.彭泽贝母组织培养和高产细胞系筛选研究[J].江西农业大学学报,2007,29(1)：152-155
[71]胡显文,高丽华,胥照平,等.稳定高产黄酮的水母雪莲细胞系的建立[J].高技术通讯,2006,16(6)：610-614
[72]梁清乐,王秋颖,曾念开,等.茯苓灭活原生质体融合育种研究[J].中草药,2006,37(11)：1733-1735
[73]王德强.红豆杉原生质体制备和培养研究[J].安徽工业大学学报,2004,21(3)：202-203,207
[74]詹忠根,徐程,张铭.铁皮石斛叶肉原生质体的分离与培养研究[J].浙江大学学报(理学版),2004,31(2)：193-196
[75]周延清,张根发,贾敬芬.影响决明无菌苗子叶原生质体分离和培养因素的研究[J].广西植物,2003,23(4)：334-338
[76]吴宝成,杭悦宇,周义锋,等.遗传育种技术在药用植物及薯蓣属植物中的应用[J].时珍国医国药,2005,16(12)：1238-1240
[77]周志军,武晓阳,孟义江,等.药用植物育种研究进展[J].时珍国医国药,2008,19(7)：1694-1698
[78]张汉明,许铁峰,等.药用植物的多倍体育种[J].中草药,2002,33(7)：UOO1-U003
[79]王彩霞.杜仲多倍体诱变育种的研究[J].内蒙古林业调查设计,2009,(1)：104-106
[80]肖艳,鲍美华,彭菲,等.紫锥菊多倍体诱导与鉴定[J].基因组学与应用生物学,2009,28(1)：149-152
[81]李鹏飞,赵晓明.牛膝多倍体的诱变及鉴定[J].山西农业大学学报(自然科学版),2009,29(2)：115-118
[82]王帅,李铂,张瑜,等.秋水仙素诱导地黄多倍体的初步研究[J].陕西农业科学,2008,54(6)：47-50
[83]李叶芳,郑思乡,关文灵,等.灯盏花多倍体植株的诱导培养研究[J].西部林业科学,2008,37(4)：71-74
[84]王跃华,胡水君,张海强,等.喜树多倍体诱导研究[J].成都大学学报(自然科学版),2008,27(3)：177-180
[85]乔永刚,赵晓明,宋芸.秋水仙素诱导黄芩多倍体的研究[J].中国医药生物技术,2008,3(5)：389-390
[86]刘丽萍,王丽艳,殷奎德.菘蓝的多倍体诱导研究[J].黑龙江八一农垦大学学报,2008,20(3)：23-26,42
[87]李文文,谢丽琼,王程,等.荒漠肉苁蓉多倍体的诱导与鉴定[J].新疆农业科学,2008,45(2)：337-341
[88]张蜀敏,王晓军,郝秀英,等.新疆雪莲多倍体的诱导初探[J].西北农业学报,2008,17(1)：216-220
[89]刘治国.宁夏育出枸杞新品系99-3[J].专业户,2001,(3)：30
[90]周延清,牛敬媛,郝瑞文,等.发根农杆菌转化怀地黄再生植株[J].分子细胞生物学报,2007,40(4)：223-231
[91]赵寿经,李昌禹,钱延春,等.人参发根的诱导及其适宜培养条件的研究[J].生物工程学报,2004,20(2)：215-220
[92]贺红,韩美丽.农杆菌介导枳壳转化系统的建立及PCR鉴定[J].中草药,2003,34(2)：171-174
[93]彭云滔,唐绍清,李伯林,等.野生罗汉果遗传多样性的ISSR分析[J].生物多样性,2005,13(1)：36-42
[94]左云娟,朱培林,刘强,等.道地药材江枳壳品种遗传学关系的ISSR证据[J].中国中药杂志,2005,30(18)：1416-1419
[95]罗光明,陈岩,李霞,等.枳壳道地产区主流品种遗传多样性的ISSR分析[J].江西农业大学学报,2007,29(1)：124-128
[96]杨红兵,崔光红,詹亚华,等.湖北恩施产厚朴ISSR指纹图谱构建[J].中药材,2009,32(1)：19-22
[97]苏雪,孙坤,陈纹,等.药用植物紫花地丁的分子鉴定及其亲缘关系研究[J].中兽医医药杂志,2009,28(2)：8-10
[98]罗成,刘锦,顾蔚,等.华中五味子AFLP反应体系的建立[J].生物技术,2009,19(2)：37-40
[99]张俊,蒋桂华,敬小莉,等.我国药用植物种质资源离体保存研究进展[J].世界科学技术,2011,12(3)23
[100]张俊,蒋桂华,敬小莉,等.我国药用植物育种的最新进展[J].中国现代中药,2010,12(5)：5-9
[2]温学森,魏建和,杨世林,等.地黄种质资源的离体保存研究[J].中国中药杂志,2003,28(1)：17-20
[3]国家药典委员会.中华人民共和国药典(2010年版)一部[S].北京：化学工业出版社,2010：38
[4]贾敏如.川芎、川白芷生产质量管理规范(GAP)的研究[M].成都：四川科学技术出版社,2007
[5]王昌华,刘翔,银福军,等.大黄种子质量分级标准的研究[J].时珍国医国药2009,20(7)：1605-1606
[6]孙群,杨力钢,丁自勉,等.乌拉尔甘草种子质量分级标准的研究[J].中国中药杂志,2008,33(10)：1126-1129
[7]贺玉林,李先恩,淡红梅.远志种子质量分级标准研究[J].种子,2007,26(1)：106-107
[8]贺玉林,李先恩,淡红梅,等.远志种子检验规程[J].中国中药杂志,2007,32(15)：1497-1500
[9]吾拉尔古丽,王建华,李先恩.板蓝根种子发芽试验标准化研究[J].种子,2005,24(6)：34-36
[10]胡榜文,薛江涛,李国峰,等.玄参脱毒种苗生产标准操作规程[J].现代中药研究与实践,2008,22(6)：17-19
[11]陈翠,康平德,杨丽云,等.云南重楼种苗繁育技术[J].中国现代中药,2010,12(2)：23-24,28
[12]史永忠,潘瑞炽,王小菁,等.铁皮石斛种质室温离体保存[J].华南师范大学学报,1999,(4)：73-77
[13]黄菊辉,陈世儒.生姜种质资源离体保存[J].西南农业大学学报,1996,13(3)：310-312
[14]李海兵,周娜,赵姣,等.怀山药种质资源的包埋玻璃化超低温保存与植株再生[J].植物学报,2010.45(3)：379-383
[15]孔令婕,巢建国,谷巍,等.茅苍术种质资源的超低温保存[J].南京中医药大学学报,2010,26(1)：56-58
[16]陈辉,陈晓玲,陈龙清,等.百合种质资源限制生长法保存研究[J].园艺学报,2006,33(4)：789-793
[17]雷志强,张寿文,刘华,等.车前种子种苗的分级标准研究[J].江西中医学院学报,2007,19(5)：65-67
[18]张春庆,王建华.种子检验学[M].北京：高等教育出版社,2005
[19]陈火英.现代种子种苗学[M].上海：上海科学技术出版社,1999
[20]洪森荣,郭连金.离体保存技术在植物种质资源保存中的应用[J].上饶师范学院学报,2006,26(3)：92-95
[21]张巧仙,范冬梅.植物种质资源的保存——离体保存.太原师范学院学报(自然科学版[J].2005,4(3)：82-85
[22]陈晓明,刘海龙,蔡玲.桉树种质资源离体保存技术[J].广西林业科学,2009,38(1)：8-13
[23]胡炜,孙拥军.强化对种子质量及指标体系的认识[J].种子科技,2003,(4)：213
[24]黄勇.石斛属植物种质资源离体保存.湖北农业科技[J].2009,48(11)：2716-2718
[25]黄璐琦.分子生药学[M].北京：北京医科大学出版社,2000
[26]Jarret R L,Florlowski W J. In Vitro Active Vs.Field Genbank Maintenance of Sweet Potato Germplasm:Major Costs and Considerrations [J]. Hort Science, 1990,25(2):141
[27]董玉琛.保护作物多样性,发掘有用新基因——试论我国作物种质资源研究当前的任务[J].植物遗传资源科学,2000.1(1)：2
[28]马小军,肖培根.种质资源遗传多样性在药用植物开发中的重要意义[J].中国中药杂志,1998,23(10)：579
[29]周逊,向长萍.植物种质资源缓慢生长离体保存研究进展[J].中国蔬菜,2008,(11)39-41
[30]伊华林,邓秀清.植物种质离体保存技术研究进展[J].植物学通报,1999,16(5)：574-581
[31]付传明,赵志国,黄宁珍,等.药用植物红根草种质资源的离体保存[J].广西植物,2007,27(4)：653-657
[32]刘小莉,杨耀文,钱子刚.多效唑和蔗糖对金铁锁离体保存的影响[J].云南中医学院学报,2009,34-36
[33]李明军,徐鑫,夏民,等.PP333和BA组合对怀地黄试管苗生长发育的影响[J].植物学通报,2006,23(1)：56-59
[34]李峰,付传明,黄宁珍,等.巴戟天种质离体保存研究[J].广西植物,2008,28(1)：95-99
[35]Wang JF,Liu YX,Lin SQ.A study of the preservation in vitro of loquat germplasm Ⅱ:Effect of plant growth inhibitor[J].Subtrop Plant Sci,2002,31(4):1-4
[36]Kartha K K,Mroginski L A,Pahl K K,et al.Gernplassm preservation of coffea (Coffea arabica)by in vitro curure of shoot apical meristems[J].Plant Science Letters,1981,22:411-413
[37]Bertrand Desbnutais A,Noirot M,Charrier A.Slow growth invitro conservation of coffee (Coffea spp) [J]. Plant Cell,Tissue and Organ Culture,1992,31:105-110
[38]张玉进,张兴国,刘佩英.魔芋不定芽的低温保存研究[J].西南农业大学学报,1999,21(4)：303-306
[39]张玉进,张兴国,刘佩英.魔芋花粉的低温和超低温保存[J].园艺学报,2000,27(2)：139-140
[40]徐刚标,何方,陈良昌.银杏种质离体保存的研究[J].经济林研究,1999,17(4)：15-18
[41]徐刚标,何方,黄晓光.银杏种质离体保存的研究Ⅰ.银杏花粉贮存[J].中南林学院学报,2000,20(1)：27-30
[42]刘剑锋,阎秀峰,程云清,等.高山红景天(Rhodiola sachalinensis)试管苗缓慢生长法保存及试管苗DNA含量分析[J].浙江大学学报(农业与生命科学版),2007,33(4)：373-378
[44]刘小莉,杨耀文,钱子刚.多效唑和蔗糖对金铁锁离体保存的影响[J].云 南中医学院学报,2009,32(1)：34-36
[45]李锋,付传明,黄宁珍,等.巴戟天种质离体保存研究[J].广西植物,2008,28(1)：95-99
[46]付传明,黄宁珍,赵志国,等.广西地不容种质离体保存技术研究[J].广西科学,2007,14(2)：155-159
[47]史永忠,潘瑞炽,王小菁,等.铁皮石斛种质室温离体保存[J].华南师范大学学报(自然科学版),1999,(4)：73-77
[48]张晓丽,刘文英,张楠,等.PP333对怀地黄种质离体保存的影响[J].河南师范大学学报(自然科学版),2009,(3)：171-174
[49]唐凤鸾,韦记青,蒋运生,等.黄花蒿组培快繁与种质离体保存的研究[J].热带亚热带植物学报,2008,16(5)：486-490
[50]付传明,黄宁珍,唐凤鸾,等.山银花的组培快繁和种质离体保存研究[J].广西科学,2008,15(3)：304-308
[51]洪森荣,尹明华,柯维忠,等.PP333对野葛离体保存的影响[J].上饶师范学院学报,2007,27(2)：68-72
[52]张玉进,张若国,庞杰,等.魔芋茎尖玻璃化冻存研究[J].作物学报,2001,27(1)：97-102
[53]徐刚标,易文,何方,等.银杏种质离体保存的研究：Ⅱ.银杏胚超低温保存[J].中南林学院学报,2000,20(2)：7-10
[54]曾华兰,叶鹏盛,倪国成,等.川芎主要病虫害及其发生危害规律研究[J].西南农业学报,2009,22(1)：99-101
[55]张萃蓉,杨星勇,张玉芳,等.川芎块茎腐烂病研究[J].云南农业大学学报,1993,8(3)：259-261
[56]张俊,蒋桂华,敬小莉,等.我国药用植物种质资源离体保存研究进展[J].世界科学技术,2011,12(3)23
[57]张俊,蒋桂华,敬小莉,等.我国药用植物育种的最新进展[J].中国现代中药,2010,12(5)：5-9
[58]高农,韩学俭.川芎苓子繁育技术[J].特种经济动植物,2004,10：26
[59]王岚,肖海波,马逾英.川芎道地性的ISSR分析[J].四川大学学报(自 然科学版),2008,45(6)：1472-1476
[60]张丽萍,杨世林,杨春清,等.我国药材种子种苗产业存在的问题及其对策[J].中国中药杂志,1999,24(10)：579-581
[61]张廷红,郭永杰,当归种苗繁殖技术[J].甘肃农业科技,1997,8：14-15
[62]崔秀明,王朝梁,陈中坚.种苗分级对三七生长和产量的影响[J].中药材,1998,21(2)：60-61
[63]朱贞丽,王鼎峰,李勇,等.马蓝优质种苗培养技术的研究[J].海峡药学,2009,21(4)：87-88
[64]田伟,温春秀,周巧梅,等.不同来源远志种子的质量比较[J].现代中药研究与实践,2006,20(5)：19-20
[65]第九届全国人民代表大会常务委员会第十六次会议通过.中华人民共和国种子法.2000,7
[66]洪森荣,李明军.玻璃化法超低温保存怀山药种质的技术研究[J].中草药,2006,37(11)：1715-1718
[67]刘剑锋,程云清,秦小伟.高山红景天茎尖的包埋-玻璃化超低温保存研究[J].,吉林师范大学学报(自然科学版),2009,3：61-65
[68]Leunufina S,Keller E R J.Investigating a new cryopreservation protocol for yams (Diooscorea spp.) [J]. Plant Cell Reports,2003,21 (12):1159-1166
[69]Shibili R A,Moges A D,Karam N S.Cryopreservation of Africa violet (Sain tpaulia Ionantha wendl.) shoot tips [J]. In Vitro Cell Dev Biol Plant,2004,40(4):389-395
[70]Hirai D,Sakai A,Cryopreservation of shoot tips from in vitro plants of sweet potata[Ipomoea botatas (L.)Lam.] by vitrification [J]. Plant Cell Reports,2003,19(7):733-737
[71]Liu Y G, Wang X Y.Study of cyropreservation technique of apple germplasm by vitrification [J].J Shandong Agric Univ:Nat Sci, 2002,33(1):32-36
[72]Matsumoto T,Sakai A,Takahashi C.Cryopreservation of in vitro-grown spical meristems of wasabi (Wasabia japonica) by encapsulation-vitrifacation method[J].Cryo-Letters,1995,16(4):189-196
[73]Matsumoto T,Sakai A,Yamada K.Cryopreservation of in vitro-grown spical meristems of wasabi (Wasabia japonica) by vitrifacation and subsequent high plant regeneration [J]. Plant Cells Reports,1994,13 (8):442-446
[74]Gupta S, Reed B M.Cryopreservation pf shoot tips of blackberry and raspberry by encapsulation-dehudtation and vitrification [J]. Cryo-letters,2006, 27(1):29-42
[75]张志珂,陈彬,杨凤玲,等.枇杷野生种的种质资源离体保存初探[J].复件果树,2008,3：28-32
[76]刘小军.枇杷种质离体保存的初步研究[D].福建：福建农林大学,2002
[77]Zheng S Q.Achievement and Prospect of Loquat Breeding in China[J].Acta Horticulturae,2007,705:85-92
[78]吴卫,郑有良,陈黎,等.川产鱼腥草种质资源的同工酶分析.中草药,2002,25(10)：695-698
[79]王永炎.有关中药材GAP中种质资源的若干问题剖析.第四届北京国际周论坛报告集
[80]中国药材GAP网,首页>GAP研究与实践>药用植物栽培技术.药用植物栽培(四)--药用植物的良种选育
[81]陈兴福,丁德蓉,黄文秀,等.川芎生长发育特性的研究[J].中国中药杂志,1997,22(9)：527-529
[82]黄璐琦,郭兰萍,崔光红,等.中药资源可持续利用的基础理论研究[J].中药研究与信息,2005,7(8)：4-7
[83]蒋桂华,马逾英,侯嘉,等.川芎种质资源的调查收集与保存研究[J].中草药,2008,39(4)：601-604
[84]郭才,霍志军.植物遗传育种及种苗繁育[M].北京：中国农业大学出版社,2006
[85]胡延吉.植物育种学[M].北京：高等教育出版社,2003
[86]张春庆,王建华.种子检验学[M].北京：高等教育出版社,2005
[87]喻方圆,周景莉.林木种苗质量检验技术[M].北京：中国林业出版社,2008
[1]贾敏如.川芎、川白芷生产质量管理规范(GAP)的研究[M].成都：四川科学技术出版社,2007：1
[2]万德光,彭成,赵军宁.四川道地中药材志[M].成都：四川科学技术出版社,2005：4
[3]苏颂.图经本草(胡乃长,王致谱辑注)[M].福建：福建科学技术出版社,1987：129
[4]刘圆,贾敏如.川芎品种、产地的历史考证[J].中药材.2001：24(5)：365
[5]彭县县志办公室.彭县县志(1910-1986)[M].成都：四川人民出版社,1986
[6]郭平,李祖伦,陈红,等.川芎地上部分挥发油化学成分的研究[J].中国中药杂志,1993,18(9)：551-552
[7]任东春,杨念云,钱士辉,等.川芎地上部分化学成分研究[J].中国中药杂志,2007,32(14)：1418-1420
[8]刘友平,李祖伦,张廷模,等.薄层扫描法测定川芎不同部位阿魏酸的含量[J].中国中药杂志,1995,20(1)：9-10
[9]徐杰军.常用中药药理与临床[M].成都：四川科学技术出版社.2005：25-26
[10]凌婧.川芎挥发油对大鼠脑缺血再灌注损伤的保护作用及其机制研究,四川大学硕士论.文,2006年(2003级)
[11]胡荣.川芎挥发油解热作用及机理研究,成都中医药大学硕士论文,2001年4月
[12]蒋桂华,马逾英,侯嘉,等.川芎种质资源的调查收集与保存研究[J].中草药,2008,39(4)：601-604
[13]王岚,肖海波,马逾英,等.川芎道地性的ISSR分析[J].四川大学学报(自然科学版),2008,45(6)：1472-1476
[14]赵勇,傅体华,范巧佳.川芎各器官的生长动态分析[J].西南农业学报.2008,21(4)：
[15]张毅,范巧佳,郑顺林,等.春季追肥对川芎产量、阿魏酸和总生物碱含量的影响[J].中国中医药杂志.2008,33(16)：1944-1947
[16]冯茜,黄云,巩春梅,等.川芎根腐病菌(Fusarium solani)的生物学特性[J].四川农业大学学报.2008,26(1)：24-27
[17]冯茜,何苗,黄云,等.川芎根腐病的症状及病原鉴定[J].植物保护学报.2008,35(4)：377-378
[18]陈要臻,王岚,马逾英,等.川芎简单序列重复间区多态性和扩增片段长度多态性分子 标记技术多态性比较研究[J].时珍国医国药,2009,20(6)：1422-1424
[19]马玎,马逾英,张利,等.三个不同产地的川芎与其近缘植物藁本的挥发油成分对比分析[J].中国现代中药.2009,11(7)：20-22,25
[20]王妙闻,张艺,张静,等.HPLC测定川芎中的总阿魏酸[J].华西药学杂志,2008,23(1)：100-102
[21]侯嘉,马逾英,贾敏如,等.川产道地药材川芎与云南、甘肃产川芎形态组织学研究[J].湖南中医药大学学报,2007,27(S1)：9
[22]蒋卫东,唐琳,陈鹏等. 川芎的组织培养研究. [J]安徽农业科学,2007,35(27)：8448-8450
[23]蒋卫东,唐琳,杨苍劲,等.卡拉霉素对川芎愈伤组织诱导和生长的影响[J].河南农业科学,2007(11)：85-87
[24]詹玉莲,马小军,戴均贵,等.川芎愈伤组织及悬浮培养物化学成分气相色谱-质谱联用分析[J].时珍国医国药, 2006,17(6)：1117-1119
[25]刘玉萍,曹晖,韩桂茹,等.中日产川芎的matK. ITS基因序列及其物种间的亲缘关系[J].药学学报,2002,37(1)：63-68
[26]魏建和,陈士林,程惠珍,等.中药材种子种苗标准化工程[J].世界科学技术-中医药现代化,2005,7(6)：104-108
[27]崔秀明,王朝梁,陈中坚.种苗分级对三七生长和产量的影响[J].中药材,1998,21(2)：60-61
[28]张国荣,张玉进,李生彬.乌拉尔甘草种子种苗分级标准制定[J].现代中药研究与实践,2004,18(4)：14-16
[29]张廷红,郭永杰,当归种苗繁殖技术[J].甘肃农业科技,1997,8：14-15
[30]朱贞丽,王鼎峰,李勇,等.马蓝优质种苗培养技术的研究[J].海峡药学,2009,21(4)：87-88
[31]Lyndsey A, Withers A. In vitro approaches to conversation of plant genetic resources [A]. In:Plant Tissue Culture and Its Agricultural Applications (LyndscyA. Withers and P. G. Aldrrsoneds) [C], London:Butterworths,1986:261-276
[32]Hawkes J G.Germplasm collection, preservation and use [A]. In:Plant Breeding[CKenneth J. Freg ed.]. [C]. New Delhi:Gayatri Offsite Press 1981:57-84
[33]史永忠,潘瑞炽,王小菁,等.铁皮石斛种质室温离体保存[J].华南师范大学学报(自然科学版),1999,(4)：73-77
[34]唐凤鸾,韦记青,蒋运生,等.黄花蒿组培快繁与种质离体保存的研究[J].热带亚热带植物学报,2008,16(5)：486-490
[35]李海兵,周娜,赵姣,等.怀山药种质资源的包埋玻璃化超低温保存与植株再生[J].植物学报,2010.45(3)：379-383
[36]孔令婕,巢建国,谷巍,等.茅苍术种质资源的超低温保存[J].南京中医药大学学报,2010,26(1)：56-58
[37]陈辉,陈晓玲,陈龙清,等.百合种质资源限制生长法保存研究[J].园艺学报,2006,33(4)：789-793
[38]洪森荣,郭连金.离体保存技术在植物种质资源保存中的应用[J].上饶师范学院学报,2006,26(3)：92-95
[39]Mantell S H, Smith H. Plant biotechnology[M]. Cambridge:Cambridge University Press,1983.187-218
[40]张玉进,张若国,庞杰,等.魔芋茎尖玻璃化冻存研究[J].作物学报,2001,27(1)：97-102
[41]徐刚标,何方,黄晓光.银杏种质离体保存的研究：Ⅰ.银杏花粉贮存[J].中南林学院学报,2000,20(1)：27-30
[42]徐刚标,易文,何方,等.银杏种质离体保存的研究：Ⅱ.银杏胚超低温保存[J].中南林学院学报,2000,20(2)：7-10
[43]高山林.药用植物遗传育种的现状与展望[J].世界科学技术：中药现代化,2001,3(6)：58-62
[44]徐昭玺,魏建和,冯秀娟,等.边条人参新品种的系统选育[J].中国医学科学院学报,2001,23(6)：542-546
[45]朱培林,吴永忠,兰冬生,等.江枳壳优良类型选择研究[J].现代中药研究与实践,2004,18(6)：24-26
[46]何先元,郭巧生,徐文斌,等.不同药用白菊花栽培品种田间试验研究[J].安徽农业大学学报,2005,32(3)：385-388
[47]王秋颖,郭顺星.天麻优良品种选育的初步研究[J].中国中药杂志,2001,26(11)：744-746
[48]王锦秀,赵健,黄占明.枸杞与番茄属间远缘杂交研究初报[J].宁夏农林科技,2005,(3)：8-9
[49]吴才祥,杨晟永,葛芝富.天麻远缘杂交育种初报[J].湖南林业科技,2007,34(1)：23-25
[50]唐宁.白术辐射诱变育种研究[J].安徽农学通报,2007,13(23)：84-85
[51]唐宁,张敦林.白术辐射诱变株系抗氧化酶活性研究[J].安徽农学通报,2007,13(24)：54-55
[52]唐宁.白术辐射诱变株系中白术内酯Ⅰ、Ⅲ的含量测定[J].安徽农学通报,2009,15(1)：45-45,19
[53]颉红梅,刘效瑞,李文建,等.甘肃当归新品系DGA2000-02的选育研究[J].原子核物理评论,2008,25(2)：196-200
[54]甄卫军,孙月华,马凤云.低能离子注入麻黄种子生物效应初步研究[J].种子,2003,(2)：26-27,44
[55]潘燕,肖翔,吴李君,等.N+离子注入对干旱胁迫条件下甘草幼苗的SOD酶和CAT酶活性及丙二醛含量的影响[J].激光生物学报,2005,14(6)：442--446
[56]魏胜林,吴李君,谢传晓,等.N+注入对甘草叶片腺体和腺体分泌多糖及叶片多糖的影响[J].核技术,2008,25(2)：196-200
[57]李耀维,武增寿.激光诱变选育雷公藤次生物质高产细胞系[J].激光生物学报,2000,9(4)：281-284
[58]郭斌,尉亚辉,等He-Ne激光诱变选育高产白藜芦醇细胞系[J].光子学报,2002,31(3)：277-280
[59]单成钢,王志芬,苏学合,等.航天诱变黄芩种子对其SP1代的影响[J].核农学报,2008,22(2)：188-191
[60]王志芬,苏学合,闫树林,等.太空搭载桔梗种子SP1代的生物学效应研究[J].核农学报,2004,18(4)：323-324
[61]王志芬,单成钢,苏学合,等.丹参种子航天搭载的诱变效应研究[J].现代中药研究与实践,2007,21(4)：6-8
[62]李金贵,朱奎,沈海玉,等.卫星搭载牛膝种子SP1植株的生物学特性[J].中国农学通报,2007,23(7)：42-45
[63]熊大胜,朱金桃,张自亮,等.三叶木通理化诱变技术及其成熟期变异研究[J].常德师范学院学报(自然科学版),2001,13(4)：79-80,89
[64]张秀省,张荣涛,曹岚,等.EMS诱变的长春花细胞系突变研究[J].中草药,2004,25(11)：1293-1296
[65]孔青,刘林,李华斌.药用植物女贞的快繁[J].山东林业科技,2008,38(3)：69-70
[66]瞿大枞,陈崇顺,王轶,等.药用植物半枝莲的离体快繁[J].安徽农业科学,2007,35 (17)：5065-5066,5095
[67]唐伟斌,石晓云.药用植物桔梗组培快繁体系的建立[J].河北师范大学学报(自然科学版),2006,30(5)：599-601
[68]刘建成,陈先玉.川西獐牙菜的组织培养与快速繁殖[J].植物生理学通讯,2003,39(3)： 237-237
[69]温春秀,谢晓亮,吴志明,等.丹参脱病毒及组培快繁技术研究[J].中草药,2004,35(9)：1057-1059
[70]张寿文,雷志强,刘华,等.彭泽贝母组织培养和高产细胞系筛选研究[J].江西农业大学学报,2007,29(1)：152-155
[71]胡显文,高丽华,胥照平,等.稳定高产黄酮的水母雪莲细胞系的建立[J].高技术通讯,2006,16(6)：610-614
[72]梁清乐,王秋颖,曾念开,等.茯苓灭活原生质体融合育种研究[J].中草药,2006,37(11)：1733-1735
[73]王德强.红豆杉原生质体制备和培养研究[J].安徽工业大学学报,2004,21(3)：202-203,207
[74]詹忠根,徐程,张铭.铁皮石斛叶肉原生质体的分离与培养研究[J].浙江大学学报(理学版),2004,31(2)：193-196
[75]周延清,张根发,贾敬芬.影响决明无菌苗子叶原生质体分离和培养因素的研究[J].广西植物,2003,23(4)：334-338
[76]吴宝成,杭悦宇,周义锋,等.遗传育种技术在药用植物及薯蓣属植物中的应用[J].时珍国医国药,2005,16(12)：1238-1240
[77]周志军,武晓阳,孟义江,等.药用植物育种研究进展[J].时珍国医国药,2008,19(7)：1694-1698
[78]张汉明,许铁峰,等.药用植物的多倍体育种[J].中草药,2002,33(7)：UOO1-U003
[79]王彩霞.杜仲多倍体诱变育种的研究[J].内蒙古林业调查设计,2009,(1)：104-106
[80]肖艳,鲍美华,彭菲,等.紫锥菊多倍体诱导与鉴定[J].基因组学与应用生物学,2009,28(1)：149-152
[81]李鹏飞,赵晓明.牛膝多倍体的诱变及鉴定[J].山西农业大学学报(自然科学版),2009,29(2)：115-118
[82]王帅,李铂,张瑜,等.秋水仙素诱导地黄多倍体的初步研究[J].陕西农业科学,2008,54(6)：47-50
[83]李叶芳,郑思乡,关文灵,等.灯盏花多倍体植株的诱导培养研究[J].西部林业科学,2008,37(4)：71-74
[84]王跃华,胡水君,张海强,等.喜树多倍体诱导研究[J].成都大学学报(自然科学版),2008,27(3)：177-180
[85]乔永刚,赵晓明,宋芸.秋水仙素诱导黄芩多倍体的研究[J].中国医药生物技术,2008,3(5)：389-390
[86]刘丽萍,王丽艳,殷奎德.菘蓝的多倍体诱导研究[J].黑龙江八一农垦大学学报,2008,20(3)：23-26,42
[87]李文文,谢丽琼,王程,等.荒漠肉苁蓉多倍体的诱导与鉴定[J].新疆农业科学,2008,45(2)：337-341
[88]张蜀敏,王晓军,郝秀英,等.新疆雪莲多倍体的诱导初探[J].西北农业学报,2008,17(1)：216-220
[89]刘治国.宁夏育出枸杞新品系99-3[J].专业户,2001,(3)：30
[90]周延清,牛敬媛,郝瑞文,等.发根农杆菌转化怀地黄再生植株[J].分子细胞生物学报,2007,40(4)：223-231
[91]赵寿经,李昌禹,钱延春,等.人参发根的诱导及其适宜培养条件的研究[J].生物工程学报,2004,20(2)：215-220
[92]贺红,韩美丽.农杆菌介导枳壳转化系统的建立及PCR鉴定[J].中草药,2003,34(2)：171-174
[93]彭云滔,唐绍清,李伯林,等.野生罗汉果遗传多样性的ISSR分析[J].生物多样性,2005,13(1)：36-42
[94]左云娟,朱培林,刘强,等.道地药材江枳壳品种遗传学关系的ISSR证据[J].中国中药杂志,2005,30(18)：1416-1419
[95]罗光明,陈岩,李霞,等.枳壳道地产区主流品种遗传多样性的ISSR分析[J].江西农业大学学报,2007,29(1)：124-128
[96]杨红兵,崔光红,詹亚华,等.湖北恩施产厚朴ISSR指纹图谱构建[J].中药材,2009,32(1)：19-22
[97]苏雪,孙坤,陈纹,等.药用植物紫花地丁的分子鉴定及其亲缘关系研究[J].中兽医医药杂志,2009,28(2)：8-10
[98]罗成,刘锦,顾蔚,等.华中五味子AFLP反应体系的建立[J].生物技术,2009,19(2)：37-40
[99]张俊,蒋桂华,敬小莉,等.我国药用植物种质资源离体保存研究进展[J].世界科学技术,2011,12(3)23
[100]张俊,蒋桂华,敬小莉,等.我国药用植物育种的最新进展[J].中国现代中药,2010,12(5)：5-9