1. 毕业设计（论文）的内容和要求

1．文献查阅 查阅间歇浸没反应器(tibs)、金钗石斛原球茎培养培养以及石斛碱(dendrobine)检测方法的相关文献。

2．阅读文献及完成开题 阅读相关文献，了解金钗石斛原球茎培养的研究现状以及非生物与生物因子对石斛碱生物合成的影响，完成开题报告。

3．明确实验任务，制定实验计划 根据所查阅文献的相关内容，确定实验内容及方案，制定科学可行的研究计划。

2. 参考文献

[1] 张露月. 金钗石斛总黄酮与石斛碱的提取分离及总黄酮抗氧化活性研究[D]. 江南大学, 2016. [2] 张晓敏, 孙志蓉, 陈龙,等. 金钗石斛的化学成分和药理作用研究进展[J]. 中国现代应用药学, 2014, 31(7):895-899. [3] 张宇斌, 乙引, 陈训,等. 铁皮石斛原胚体高效诱导、增殖及多糖含量累积的研究[J]. 吉林农业, 2012(8):46-47. [4] Lu S X, He T G, Wang A Q, et al. Study on extraction method of chemical components of Dendrobium candidum.[J]. Southwest China Journal of Agricultural Sciences, 2010, 23(2):523-527. [5] Luo C, He T, Chun Z. Discrimination and chemical phylogenetic study of seven species of Dendrobium, using infrared spectroscopy combined with cluster analysis[J]. Journal of Molecular Structure, 2013, 1037(14):40-48. [6] Sukphan P, Sritularak B, Mekboonsonglarp W, et al. Chemical constituents of Dendrobium venustum and their antimalarial and anti-herpetic properties[J]. Natural Product Communications, 2014, 9(6):825-827. [7] Chen X, Zhang S, Ma J, et al. A simple, rapid and reliable UFLC-MS/MS method for the determination of dendrobine in rat plasma and its application to a pharmacokinetic study[J]. Analytical Methods, 2014, 6(4):1197-1202. [8] Chen K K, Chen A L. The pharmacological action of dendrobine. The alkaloid of Chin-shih-hu[J]. Journal of Pharmacology Experimental Therapeutics, 1935, 55(3):319-325. [9] Onaka T, Kamata S, Maeda T, et al. THE STRUCTURE OF DENDROBINE.[J]. Chemical Pharmaceutical Bulletin, 1964, 12(12):506-512. [10] Glomqvist L, Brandauml;nge S, Gawell L, et al. Studies on orchidaceae alkaloids. XXXVII. Dendrowardine, a quaternary alkaloid from Dendrobium wardianum Wr[J]. Acta Chemica Scandinavica, 1973, 27(4):1439. [11] Hiroshi Morita, Masako Fujiwara, Naotoshi Yoshida,等. New Picrotoxinin-type and Dendrobine-type Sesquiterpenoids from Dendrobium, Snowflake ”Red Star#8217;[J]. Tetrahedron, 2000, 56(32):5801-5805. [12] 张光浓，毕志明，王峥涛等. 石斛属植物化学成分研究进展[J]. 中草药，2003，34(6): 5-8 [13] 金蓉鸾，孙继军，张远名. 11种石斛的总生物碱的测定[J]. 中国药科大学学报，1981(1): 9-13 [14] Wang Y H, Avula B, Abe N, et al. Structural characterization of dendrobine-type sesquiterpene alkaloids from the stems of Dendrobium nobile using LC-MS/MS[C]// Meeting of the American-Society-Of-Pharmacognosy. 2014:802-802. [15] 丁亚平, 杨道麒, 吴庆生,等. 安徽霍山三种石斛总生物碱的测定及其分布规律研究[J]. 安徽农业大学学报, 1994(4):503-506. [16] 罗建平, 查学强, 姜绍通. 药用霍山石斛原球茎的液体悬浮培养[J]. 中国中药杂志, 2003, 28(7):611-614. [17] 刘进平, 莫饶. 热带植物组织培养[M]. 科学出版社, 2006. [18] 查学强, 罗建平, 石玮,等. 金属离子对霍山石斛类原球茎增殖及植株再生的影响[J]. 园艺学报, 2006, 33(1):179-181. [19] 蒋海侠. 铁皮石斛原球茎的扩繁及其与内生菌共培养[D]. 南京工业大学, 2016. [20] 涂红艳, 刘伟, 叶庆生. 细茎石斛拟原球茎生长与有效成分积累的关系[J]. 现代生物医学进展, 2004, 4(3):37-41. [21] Kutney J P, Samija M D, Hewitt G M, et al. Anti-inflammatory oleanane triterpenes from Tripterygium wilfordii, cell suspension cultures by fungal elicitation[J]. Plant Cell Reports, 1993, 12(6):356. [22] Nef C,Trouslot MF,Trouslot P,etal. Long-term effeet of Pythium elieitor treatmenton the growth and alkaloi Produetion of Catharanthus roseus ceellsus Pensions[J],PlantMed,1994,60:149-152. [23] Singh G J, Oakey J S, Curtis W R. Interaction of methyl jasmonate, wounding and fungal elicitation during sesquiterpene induction in Hyoscyamus muticus in root cultures[J]. Plant Cell Reports, 1998, 17(5):391-395. [24] 胡燕花，张本厚，贾明良，陈集双，间歇浸没式生物反应器培养铁皮石斛组培苗，中国农业科技导报, 2016 , 18 (3) :190-194 [25] 赵望锋，王力华，间歇浸没式生物反应器在大规模组织培养中的应用研究，安徽农业科学, 2007 , 35 (2) :317-320 [26] Song JY, Zhang YL,Qi JJ. Biotechnology of Salvia miltiorrhiza[J]. NatUral Prod-uct Research and Development,1998,7(20):86-89. [27] CuiTB, GuoY, LinWT. The ways to enhance the Produetion of sceondary metab-olites by Plant cell culture[J]. Plant Physiology Communications, 2001, 37(5):479-482. [28] Guerrero M F, Puebla P, Mart#237;n M L, et al. Inhibitory effect of N(G)-nitro-L-arginine methyl ester on the anti-adrenergic response elicited by ayanin in the pithed rat.[J]. Planta Medica, 2002, 68(04):322-325. [29] 陈晓梅,郭顺星.4种内生真菌对金钗石斛无菌苗生长及其多糖和总生物碱含量的影响[J].中国中药杂志,2005(04):14-18. [30] 宋经元,郭顺星,肖培根.氮源和真菌诱导子对铁皮石斛原球茎悬浮培养的影响[J].云南植物研究,2008(01):105-109. [31] 张长平, 李春, 元英进,等.真菌诱导子对悬浮培养南方红豆杉细胞态势及紫杉醇合成的影响[J].生物工程学报,2001,17(4):436-440

3. 毕业设计（论文）进程安排

2018.12.20-2019.1.13：确定课题，查找文献，提交开题报告 2019.2.20-2019.3.15：金钗石斛原球茎的增殖培养 2019.3.16-2019.4.16：设计两种不同非生物诱导子单因素实验探究其对石斛碱积累的影响； 2019.5.18-2019.5.30：撰写毕业论文，准备论文答辩