臺灣博碩士論文加值系統

春石斛蘭(Nobile-type Dendrobium)是新興蘭科作物之一，並在出口市場上具有經濟價值和外銷潛力。本實驗室以30 MS培養基(30 g/L蔗糖+0.4 mg/L BA + 0.1 mg/L NAA 之MS培養基)對多個春石斛品系進行繼代培養後，部分品系的小植株呈現矮小或黃褐化等情形。因此，本研究目的以30 MS培養基為基礎，挑選12個春石斛蘭品系之莖節培養並改良培養基配方；調整大量元素濃度、使用澱粉作為替代碳源、添加有機添加物和活性碳等，改善春石斛組織培養之培養基。結果顯示，大量元素鹽濃度調整試驗，Den. ‘9013’培養於四個處理組(全量、3/4、2/4、1/4)之30MS培養基中，3/4處理組以根系發育較其他處理組為佳，植株存活率高達90%以上；而另以四個品系609 (09129)、FL-009 (11281)、NPUD-27 (10061)、NPUD-66 (11322)於兩個處理組(全量、3/4)30 MS培養基中，不定芽數、株高、葉和根數均無差異，但FL-009 (11281)的不定芽產生於3/4處理組較為快速；替代碳源試驗，Den. ‘9013’於添加10 g/L馬鈴薯澱粉+15 g/L蔗糖之3/4 MS培養基中，根系發育較另外兩組(糯米粉、蓬萊米粉)為佳；並以馬鈴薯澱粉作為替代碳源，命名為PS培養基；另以上述四個品系於有無馬鈴薯澱粉(PS、30MS)之培養基中，四個品系的整體植株發育均無差異，但於PS培養基中根系較短小、培植體無黃褐化；上述五個品系於有無有機添加物和活性碳之培養基中，均於PPS+AC培養基(1 g/L Peptone 和1 g/L AC之PS培養基)中植株發育較為佳；並依此結果，另以七個品系於PPS+AC培養基中；其中以NPUD-20 (09074-910-14)、NPUD-9 (09001-14-3)、NPUD-2 NPUD-71 (11241)植株生長發育為最佳。

The Dendrobium（Nobile-type）is a new genus of orchids, and it has great potential to be economically significant and important in the export market. In our laboratory, Den. nobile strains were subcultured in 30 MS (MS medium with 30 g/L sucrose, 0.4 mg/L BA and 0.1 mg/L NAA ). Although this medium supported rapid growth, however, some strains showed signs of abnormal growth. Therefore, in this study, select 12 strains of Dendrobium's shoots were cultured and improved 30 MS medium recipe. Includes adjustment of macro salts strength, added starch as an alternative carbon source, as well as the addition of organic matter and activated carbon. The result in the adjustment of macro salts strength test showed that the Den. ‘9013’ cultivated in four treatments (Full, 3/4, 2/4, 1/4) of 30MS medium, 3/4 30 MS had a better root development than other treatments, and survival rate of explants reached up to 90%. Another four strains 609 (09129), FL-009 (11281), NPUD-27 (10061), NPUD-66 (11322) were cultivated in (Full, 3/4) 30 MS medium showed no significant difference in growth, with the exception of the bud regeneration of FL-009 (11281) was faster in the 3/4 treatment. In the alternative carbon source test, Den.‘9013’was cultivated in 3/4 30MS medium containing 15 g/L sucrose, and 10 g/L potato starch(PS medium), root development is better than the other two treatments (glutinous rice, japonica rice). Therefore, the use of potato starch can be an alternative carbon source. In addition, the above four strains cultivated in PS and 30 MS medium(with or without potato starch) showed no significant difference in growth, however, the explants’ roots were relatively shorter. Furthermore, there was no sign of browning in the PS medium. The above five strains were cultivated in four mediums (with or without organic matter and activated carbon, AC). The results showed that PPS+AC medium (PS medium with 1 g/L Peptone and 1 g/L AC) yielded the best growth development. Based on this result, another 7 strains were cultivated in PPS+AC medium for comparison. The growth development of NPUD-20 (09074-910-14), NPUD-9 (09001-14-3), and NPUD-2NPUD-71 (11241) were relatively better in this medium.

摘要 I
Summary II
謝誌 IV
目錄 V
圖表目錄 VIII
培養基代號 X
壹、前言 1
貳、前人研究 2
一、 石斛蘭屬之介紹 2
二、 春石斛蘭之介紹 2
(一)植株型態 2
(二)生育特性 3
(三)產業概況 3
(四)繁殖 4
三、 培養基 7
(一)氮源 7
(二)碳源 8
(三)維生素和氨基酸 9
(四)植物生長調節劑 9
(五)有機添加物 10
(六)其他物質 11
參、材料與方法 13
一、試驗材料 13
二、植株材料準備 13
(一)扦插消毒與高芽誘導 13
(二)高芽繼代 13
(三)莖節培植體 14
(四)培養基備製和培養容器 14
(五)培養室條件 14
二、不同培養基成分對春石斛品系之芽體增殖影響 14
(一) 大量元素鹽濃度調整之試驗 14
(二) 食用澱粉作為替代碳源之試驗 15
(三) 添加有機添加物之試驗 16
(四) 添加活性碳之試驗 17
四、 試驗設計和統計分析 18
肆、結果 20
一、大量元素鹽濃度調整之試驗 20
(一)春石斛‘9013’於不同大量元素鹽濃度之30 MS培養基中芽體增殖之影響 20
(二) 四個春石斛品系於兩種大量元素鹽濃度(4/4、3/4)之30 MS培養基中芽體增殖之比較 20
二、食用澱粉作為替代碳源之試驗 21
(一)春石斛‘9013’於不同馬鈴薯澱粉濃度之3/4 30 MS培養基中芽體增殖之影響 21
(二)春石斛‘9013’於不同澱粉於3/4 30MS培養基中芽體增殖之影響 22
(三)四個春石斛品系於有無馬鈴薯澱粉之培養基中芽體增殖之比較 22
三、添加有機添加物之試驗 23
(一)春石斛‘9013’於不同蛋白腖濃度之PS培養基中芽體增殖之影響 23
(二)五個春石斛品系於不同有機添加物之PS培養基中芽體增殖之影響 23
四、添加活性碳之試驗 25
(一)五個春石斛品系於有無添加活性碳之PPS培養基中芽體增殖之影響25
(二)七個春石斛品系於PPS+AC培養基中芽體增殖之影響 26
伍、討論 27
陸、結論 31
柒、參考文獻 64
捌、作者簡介 75
 

王美琪、林瑞松. 2016. 培植體分生組織的褐化與壞死之因素. 興大園藝 41 (3): 75-86.
市橋正一、蔡娟婷. 2011. 日本之春石蘭花產業及基礎生理研究. 植物種苗 13(3): 1-18.
史宗義. 2012. 蘭花翻盆分株操作規程. 中國花卉園藝 2: 34-38.
何松林、孔德政、楊秋生、張啟翔、田中道男. 2003. 碳源和有機添加物對文心蘭原球莖增殖的影響. 河南農業大學學報 37(2): 154-157.
吳金濱、何惠雅、胡雯茹、廖容君、陳忠川、北中進. 2003. 石斛的組織培養. 中臺灣醫學科學雜誌 8(s_1): s22-s28.
李淑真、廖芳心、鄭隨和. 2003. 金草蘭微體繁殖技術之研究. 桃園區農業改良場研究彙報 56: 18-25.
李亮、張冬敏、雷華輝、鐘鳳林. 2012. 植物组織培養中有機添加物應用研究. 寧夏農林科技 53(2): 28-30.
李淑真、廖芳心. 2014. 培養基組成分對台灣風蘭及厚葉風蘭種子發芽與小苗生育之影響. 桃園區農業改良場研究彙報 75: 31-46.
肖善軍. 2015. 蘭花分株繁殖與盆栽護理技術. 現代農業科技 9: 165-168.
林月金、賴本智. 1991. 蘭花農場之經營管理. 台灣農業經營研討會專集 80: 225-238.
孟志霞、房慧勇、郭順星、陳曉梅、肖培根. 營養因子對鐵皮石斛幼苗生長的影養. 2008. 中國藥學染志 43(9): 665-668.
金石文、呂廷森、陳福旗, 2010. 春石斛育種及種苗生產技術之研發.花卉研究團隊研究現況與展望研討會專刊，第49-61頁. 農業試驗所特刊第154號，行政農委會，5頁.
林育聖、董怡君、王昭月、陳俊麟、曾夢蛟. 2014. 春石斛蘭品種(系)之親緣關係的研究. 興大園藝 39 (4): 67-82.
明興加、馮婷婷. 2010. 中國石斛屬植物種苗繁育技術的研究進展.安徽農業科學 38(6): 2899 -2902.
徐懷恩、林瑞松. 文心蘭生育習性之研究. 興大園藝 22(2): 123-134.
徐红、劉俊、王峥濤、徐德然、丁家宜. 2001. 鼓槌石斛组織培養研究. 中國中藥染志 26(6): 304-306.
修景潤、朴炫春、姚睿、高日、廉美蘭. 2012. 培養基種類、BA濃度和蔗糖濃度對春石斛组培苗增殖的影響. 北方園藝 12: 143-145.
郭宏遠. 2002. 世界蘭花貿易概況. 台灣花卉園藝月刊 185: 37-40.
郭孄婷、陳尚謙、陳思吟、劉明宗. 2016. 蘭花品種選育與產期調節技術開發. 行政院農委會種苗改良繁殖場年報 105: 55-57.
康國章、王永華、郭天財、朱雲集、官春雲. 2014. 植物澱粉合成中的關鍵酶. Hereditas (Beijing) 28(1): 110-116.
許嘉錦. 2014. 春石斛蘭產業發展與育種方向. 臺中區農業專訊 84: 4-7.
許嘉錦. 2015. 春石斛蘭之栽培技術春石斛蘭之栽培技術. 臺中區農業專訊 88: 4-7.
張孟錦、陳文貞、楊志娟、江秀娜、林漢銳. 2011. 春石斛生物學特性及栽培技術研究進展. 中國農業通報 27(6): 35-39.
張祥樂、蔣戰國. 2011不同品種馬鈴薯澱粉和還原糖含量測定. 中國科技論文在線網站. 2011年6月11日. 取自http://www.paper.edu.cn/.
張珈錡、廖玉珠. 2014. 種苗繁殖及栽培技術研究. 行政院農委會種苗改良繁殖場年報 102: 25-50.
曾銀位、葉茂生. 2000. 花生花藥培養的研究Ⅲ.ABA及不同種類澱粉對花藥癒合組織分化之影響. 農林學報 49(3): 25-38.
黃柄龍. 2011. 無機鹽對藥用植物欒樨試管苗生長之影響. 高雄區農業改良場研究彙報 22(1): 22-31.
游雲飛. 2018. 秋石斛莖段離體再生體系的建立. 亞熱帶農業研究 14(4): 271-275.
劉曉青、周建濤、王燕敏. 2004. 高檔盆栽蘭花春石斛栽培管理技術. 安徽農業科學 32(5): 1093-1094.
劉海、羅鳴、黄竹榮、楊琳、吳明開. 2014. 流蘇石斛種質資源保存及组培苗與高芽苗差異研究. 貴州中藥現代化科技 33(11): 61-63.
陳福旗、陳采晴. 1998. 無機鹽類濃度及有機添加物對文心蘭擬原球體及組培苗生長之影響. 中國園藝 44(4): 403-412.
陳威臣、夏奇鈮、葉茂生、蔡新聲. 2004. 鹽類濃度、蔗糖、植物生長素及培養容器瓶口覆蓋物對高氏柴胡組培苗發根與馴化之影響. 中華農業研究 53: 249-260.
陳燕. 2007. 春石斛的組織培養和快速繁殖研究. 江西農業學報 19(10): 96-97.
陳美齡、洪惠娟. 2013. MS培養基及N6-Benzylaminopurine濃度對春石斛蘭莖節培養及植株. 臺中區農業改良場研究彙報 118: 1-10.
魏芳明、洪惠娟、楊旻憲. 2011. 春石斛蘭栽培管理技術春石斛蘭栽培管理技術. 臺中區農業專訊 74: 4-6.
魏芳明. 2012. 春石斛蘭產業及研發概況. 臺中區農業改良場特刊 111:
139-143.
顏志勳、陳加忠. 2001. 組織培養瓶培養基分注機構之研發. 農林學報
50(4): 29-41.
鄭寶强、鄭茜玫、李奎、繆崑、王雁. 2017. 不同溫度處理對石斛蘭花芽分化和發育的影響. 林農科学研究 30(3): 460-464.
Atwood, J. T. 1986. The size of the Orchidaceae and systematic distribution of epiphytic orchids. Selbyana 9: 171-186.
Arditti, J., 1997. Clonal Propagation of Orchids by Means of Tissue Culture-A Manual. In Orchid Biology: Reviews and Perspectives Ⅶ, ed. J. Arditti (Cornell University Press, Ithaca, New York). p. 203-294.
Akter, S., K. M. Nasiruddin and A. B. M. Khaldun. 2007. Organogenesis of Dendrobium orchid using traditional media and organic extracts. J. Agric. Rural Develop. 5: 30-35.
Aslantas, R., R. Cakmakci and F. Sahin. 2007. Effect of plant growth promoting rhizobacteria on young apple tree growth and fruit yield under orchard conditions. Sci. Hortic. 111: 371-377.
Al-Khateeb, A. A. 2008. Regulation of in vitro bud formation of date palm (Phoenix dactylifer L.) cv. Khanezi by different carbon sources. Bioresource Technol. 99: 655-655.
Asghar, S., T. Ahmad, I. A. Hafiz and M. Yaseen 2011. In vitro propagation of orchid (Dendrobium nobile) var. Emma white. Afr. J. Biotechnol. 10: 3097-3103.
Aggarwal, S. and C. Nirmala. 2012. Utilization of coir fibers as an eco friendly substitute for costly gelling agents for in vitro orchid seed germination. Sci. Hortic. 133: 89-92.
Bensaddek, L., F. Gillet, J. E. N. Saucedo, M. A. Fliniaux. 2001. The effect of nitrate and ammonium strengths on growth and alkaloid accumulation of Atropa belladonna hairy roots. J. Biotechnol. 85: 35-40.
Bhatia, S. and R. Dahiya. 2015. Concepts and techniques of plant tissue culture science. Modern applications of plant biotechnology in pharmaceutical sciences. Academic press, Elsevier. p. 121-156.
Bhatia, S. and K. Sharma. 2015. Plant Tissue culture-based industries. In: Modern applications of plant biotechnology in pharmaceutical sciences. Academic press, Elsevier. p. 405-417.
Chen, F. C., Y. W. Huang, H. H. Wu, P. J. Wen, C. C. Hsu, J. Y. Tsao, and T. M. Shen.2009. Phalaenopsis cloning: strategy for PLB or shoots? Proc. Taiwan International Orchid Symposium 2009. pp.8-31, Taiwan Orchid Growers’Association.
Chugh, S., S. Guha and I. U. Rao. 2009. Micropropagation of orchids: A review on the potential of different explants. Sci. Hortic. 122: 507-520.
Cheruvathur, M. K., J. Abraham, B. Mani and T. D. Thomas. 2010. Adventitious shoot induction from cultured internodal explants of Malaxis acuminata D. Don, a valuable terrestrial medicinal orchid. Plant Cell Tiss. Organ Cult. 101: 163-170.
Colombo, R. C., V. Favetta and R. T. Faria. 2015. Cutting size in the vegetative propagation of Dendrobium nobile Lindl. Agron. Sci. Biotechnol. 1(2): 73-76.
Chin, C. K., Z. H. Lee, S. A. Mubbarakh, J. J. J. Antony, B. L. Chew and S. Subramaniam. 2019. Effects of plant growth regulators and activated charcoal on somaclonal variations of protocorm-like bodies (PLBs) of Dendrobium Sabin Blue orchid. Biocatal. Agric. Biotechnol. 22: 2-8.
Da Silva, J. A. T., D. T. T. Giang, M. T. Chan, Sanjaya, A. Norikane, M. L. Chai, J. C. Ruíz, S. Penna, T. Granström, M. Tanaka. 2007. The Influence of different carbon sources, photohetero-, photoauto- and photomixotrophic conditions on protocorm like body organogenesis and callus formation in thin cell layer culture of Hybrid Cymbidium (Orchidaceae). Orchid Sci. Biotechnol. 1(1): 15-23.
Dake, Z., H. Guangwan, C. Zhiying, S. Yana, Z. Li, T. Anjun and L. Chunlin. 2013. Micropropagation and in vitro flowering of Dendrobium wangliangii: A critically endangered medicinal orchid. J. Med. Plants Res. 7(28): 2098-2110.
Da Silva, J. A. T. 2013. The effect of ethylene inhibitors (AgNO3, AVG), an ethyleneliberating compound (CEPA) and aeration on the formation of protocorm-like bodies of hybrid Cymbidium (Orchidaceae). Front. Biol. 8(6): 606-610.
Da Silva, J. A. T., S. Aceto, W. Liu, H. Yu and A. Kanno. 2014. Genetic control of flower development, color and senescence of Dendrobium orchids. Sci. Hortic. 175: 74-86.
De, L. C., P. K. Rajeevan, P. Pathak. 2014. Global Orchid Industry. Bot. J. Linn. Soc. 186: 13-19.
David, D., R. Jawan, H. Marbawi and J. A. Gansau. 2015. Organic additives Improves the in vitro growth of native orchid Vanda belvola Blume. Not. Sci. Biol. 7(2): 192-197.
Da Silva, J. A., J. C. Cardoso, J. Dobranszki and S. Zeng. 2015. Dendrobium micropropagation: a review. Plant Cell Rep. 34: 671-704.
Dirks, M. A., R. Butot, P. V. Schaik, J. W. Wijnands, R.V. D. Berg, L. Krol, S. Doebar, K. V. Kooperen, H. D. Boerl, E. M. Kramer, E. F. Smets, R. A. Vos, A. Vrijdaghs and B. Gravendeel, 2017. Exploring the evolutionary origin of floral organs of Erycina pusilla, an emerging orchid model system. Evol. Biol. 17: 89-107.
Dulić, J., M. Ljubojević, V. Ognjanov, G. Barać and T. Dulić. 2019. In vitro germination and seedling development of two European orchid species, Himantoglossum jankae Somlyay, Kreutz & Óvári and Spiranthes spiralis (L.) Chevall. In Vitro Cell Dev. Biol. Plant 55: 380-391.
Dong, S. and D. M. Beckles. 2019. Dynamic changes in the starch-sugar abiotic stress response. J. Plant Physiol. 234: 80-93.
Eymar, E., J. Alegre, M. Toribio, D. L´opez-Vela. 2000. Effect of activated charcoal and 6-benzyladenine on in vitro nitrogen uptake by Lagerstroemia indica. Plant Cell, Tissue Organ Cult. 63: 57-65.
Efferth, T. 2019. Biotechnology applications of plant callus cultures. Engineering. 5: 50-59.
Gamboge, O. L., R. A. Miller and K. Ojimai. 1968. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151-158.
Gnasekaran, P., X. Rathinam, U. R. Sinniah, S. Subramaniam. 2010. A study on the use of organic additives on the protcorm-like bodies (PLBs) growth of Phalaenopsis violacea orchid. J. Phytol. 2(1): 29-33.
Guo1, B., B. H. Abbasi, A. Zeb, L. L. Xu and Y. H. Wei. 2011. Thidiazuron: A multi-dimensional plant growth regulator. Afr. J. Biotechnol. 10: 8984-9000.
Hew, C. S. 2001. Ancient Chinese orchid cultivation a fresh look at an age-old practice. Sci. Hortic. 87: 1-10.
Hwang, J. S., S. A. Lee, S. S. Hong, X. H. Han, C. Lee, S. J. Kang, D. Lee, Y. Kim , J. T. Hong, M. K. Lee and B. Y. Hwang. 2010. Phenanthrenes from Dendrobium nobile and their inhibition of the LPS-induced production of nitric oxide in macrophage RAW 264.7 cells. Bioorg. Med. Chem. Lett. 20: 3785-3787.
He, W. and C. Wei. 2017. Progress in C-type starches from different plant sources. Food Hydrocoll. 73: 162-175
Jheng, F. Y., Y. Y. Do, Y. W. Liauh, J. P. Chung and P. L. Huang. 2006. Enhancement of growth and regeneration efficiency from embryogenic callus cultures of Oncidium ‘Gower Ramsey’ by adjusting carbohydrate sources. Plant Sci. 170: 1133-1140.
Knudson K. 1946. A new nutrient solution for germination of orchid seeds. Am. Orchid Soc. Bull. 15: 214-217.
Kananont, N., R. Pichyangkura, S. Chanprame, S. Chadchawan and P. Limpanavech. 2010. Chitosan specificity for the in vitro seed germination of two Dendrobium orchids (Asparagales: Orchidaceae). Sci. Hortic. 124: 239-247.
Khatua, H., M. M. Khatun, M. S. Biswas, M. R. Kabir and M. Al-Amin. 2010. In vitro growth and development of Dendrobium hybrid orchid. Bangladesh J. Agric. Res. 35(3): 507-514.
Kaur, S. and K. K. Bhutani. 2012. Organic growth supplement stimulants for in vitro multiplication of Cymbidium pendulum (Roxb.) Sw. HortScience 39(1): 47-52.
Lopez, R. G. and E. S. Runkle. 2005. Environmental physiology of growth and flowering of orchids. HortScience 40(7): 1969-1973.
Luo, J. P., C. Wawrosch and B. Kopp. 2009. Enhanced micropropagation of Dendrobium huoshanense C.Z. Tang et S.J. Cheng through protocorm-like bodies: The effects of cytokinins, carbohydrate sources and cold pretreatment. Sci. Hortic. 123: 258-262.
Lembrechts, R., N. Ceusters, M. P. D. Proft and J. Ceusters. 2017. Sugar and starch dynamics in the medium-root-leaf system indicate possibilities to optimize plant tissue culture. Sci. Hortic. 224: 226-231.
Murashige,T. and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15: 473-497.
Martin, K. P., J. Geevarghese, D. Joseph and J. Madassery. 2005. In vitro propagation of Dendrobium hybrids using flower stalk node explants. Indian J. Exp. Biol. 43: 280-285.
Moraes L. and R. T. Faria 2005. Activated charcoal for in vitro propagation of Brazilian orchids. Acta. Hortic. 683: 383-390.
Nambiar, N., C. S.Tee and M. Maziah. 2012. Effects of organic additives and different carbohydrate sources on proliferation of protocormlike bodies in Dendrobium Alya Pink. Plant Omics J. 5(1): 10-18.
Novak, S. D., L. J. Luna and R. N. Gamage. 2014. Role of auxin in orchid development. Plant Signal. Behav. 9(10): 1-8.
Pan M.J. and J. van Staden. 1998. The use of charcoal in in vitro culture – A review. Plant Growth Regul. 26: 155-163.
Pornpienpakdee, P., R. Singhasurasak, P. Chaiyasap, R. Pichyangkura, R. Bunjongrat, S. Chadchawan and P. Limpanavech. 2010. Improving the micropropagation efficiency of hybrid Dendrobium orchids with chitosan. Sci. Hortic. 124: 490-499.
Pang, S., K. Pan, Y. Wang, W. Li, L. Zhang and Q. Chen. 2012. Floral morphology and reproductive biology of Dendrobium jiajiangense (Orchidaceae) in Mt. Fotang, southwestern China. Flora 207: 469-474.
Panwar, D., K. Ram, Harish and N. S. Shekhawat. 2012. In vitro propagation of Eulophia nuda Lindl., an endangered orchid. Sci. Hortic. 139: 46-52.
Prizão, E. C., L. D. M. Gonçalves, M. A. M. Gutierre, C. A. Mangolin and M. D. F. P. D. S. Machado. 2012. Activated charcoal and graphite for the micropropagation of Cattleya bicolor Lindl. and a orchid double-hybrid ‘BLC Pastoral Innocence’. Acta Sci. Agron. 34(2): 157-161.
Pradhan, B. M. and D. M. Bajracharya. 2014. Distribution pattern of Dendrobium swartz (orchidaceae) in Nepal. Pleione. 8(2): 311-319.
Parthibhan, S., M. V. Rao and T. S. Kumar. 2015. In vitro regeneration from protocorms in Dendrobium aqueum Lindley - An imperiled orchid. J. Genet. Eng. Biotechnol. 13: 227-233.
Saad, A. I. M. and A. M. Elshahed. 2012. Plant Tissue Culture Media. In: Recent advances in plant in vitro culture. Intech, Croatia, p. 29-40.
Sailo, N., D. Rai, L. C. De. 2014. Physiology of temperate and tropical orchids-an overview. Int. J. Sci. Res. 3: 3-8.
Shekarriz, P., M. Kafi, S. D. Deilamy, M. Mirmasoumi. 2014. Coconut water and peptone improve seed germination and protocorm like body formation of hybrid Phalaenopsis. Agric. Sci. Dev. 3(10): 317-322.
Thomas, T. D. 2008. The role of activated charcoal in plant tissue culture. Biotechnol. Adv. 26: 618-631.
Tsai, W. C., Y. Y. Hsiao, Z. J. Pan, C. S. Kuoh, W. H. Chen and H. H. Chen. 2008. The role of ethylene in orchid ovule development. Plant Sci. 175:98-105.
Tavares, A. R., J. L. M.Young, S. S. Ori, S. Kanashiro, G. P. P. Lima, R. M. Suzuki and E. P. Chu. 2012. Orchid in vitro growth as affected by nitrogen levels in the culture medium. Hortic. Bras. 30: 119-124.
Tharapan, S., C. Thepsithar, K. Obsuwa. 2014. An effect of organic supplements on stimulating growth of Dendrobium protocorms and seedlings. Int. Sch. Sci. Res. Tnnov. 8(7): 699-704.
Udomdee, W., P. J. Wen, S. W. Chin and F. C. Chen. 2015. Effect of carbon source on protocorm-like body induction, proliferation and regeneration in Dendrobium Snowflake ‘Red Star’. Acta Hortic. 107(8): 113-120.
Utami1, E. S. W., S. Hariyanto and Y. S. W. Manuhara. 2017. In vitro propagation of the endangered medicinal orchid, Dendrobium lasianthera J. J. Sm through mature seed culture. Asian Pac. J. Trop. Biomed. 7(5): 406-410.
Vacin, E. and Went, F. 1949. Some pH changes in nutrient solution. Bot. Gardens. Conserv. 110: 605-613.
Venturieri, G. A. and F. J. Pickscius. 2013. Propagation of nobile Dendrobium (Dendrobium nobile Lindl.) by cutting. Acta Sci. Agron. 35(4): 501-504.
Vichiato, M. R. D. M., M. Vichiato, M. Pasqual, F. A. Rodrigues and D. M. D. Castro. 2014. Morphological effects of induced polyploidy in Dendrobium nobile Lindl. (Orchidaceae). Crop. Breed. Appl. Biotechnol. 14: 154-159.
Winkle, S. C. V. and G. S. Pullman. 2005. Achieving desired plant growth regulator levels in liquid plant tissue culture media that include activated carbon. Plant Cell Rep. 24: 201-208.
Wang, H. Z., S. G. Feng, J. J. Lu, N. N. Shi and J. J. Liu. 2009. Phylogenetic study and molecular identification of 31 Dendrobium species using inter-simple sequence repeat (ISSR) markers. Sci. Hortic. 122: 440-447.
Winarto, B., F. Rachmawati, A. Santi and J. A. T. Da Silva. 2013. Mass propagation of Dendrobium ‘Zahra FR 62’, a new hybrid used for cut flowers, using bioreactor culture. Sci. Hortic. 161: 170-180.
Wang, J. and Z. Tang. 2014. The regulation of soluble sugars in the growth and development of plants. Bot. Res. 3: 71-76.
Yap, Y. M., C. S. Loh and B. L. Ong. 2008. Regulation of flower development in Dendrobium crumenatum by changes in carbohydrate contents, water status and cell wall metabolism. Sci. Hortic. 119: 59-66.
Yen, C. Y. T. and T. W. Starman. 2008. Effects of cooling temperature and duration on flowering of the nobile Dendrobium Orchid. HortScience 43: 1765-1769.
Zheng, L. L., Y. Lin, S. Lin and E. A. Cossis. 1992. The polyglutamate nature of plant folates. Phytochemistry 31(7): 2277-2282.
Zettler, L. W., C. J. Hofer. 1998. Propagation of the little club-spur orchid (Platanthera clavellata) by symbiotic seed germination and its ecological implications. Environ. Exp. Bot. 39: 189-195.
Zha, X. Q., J. P. Luo, S. T. Jiang and J. H. Wang. 2007. Enhancement of polysaccharide production in suspension cultures of protocorm-like bodies from Dendrobium huoshanense by optimization of medium compositions and feeding of sucrose. Process Biochem. 42: 344-351.
Zeng, S., J. Wang, K. Wu, J. A. T. Da Silva, J. Zhang and J. Duan. 2013. In vitro propagation of Paphiopedilum hangianum Perner & Gruss. Sci. Hortic. 151: 147-156.
Zahara, M., A. Datta, P. Boonkorkaew and A. Mishra. 2017. The effects of different media, sucrose concentrations and natural additives on plantlet growth of Phalaenopsis Hybrid ‘Pink’. Braz. Arch. Biol. Technol. 60: 1-15.
Zhang, B., S. Sarsaiya, X. Pan, L. Jin, D. Xu, B. Zhang, G. J. Dunsa, J. Shib, J. Chena. 2018. Optimization of nutritional conditions using a temporary immersion bioreactor system for the growth of Bletilla striata pseudobulbs and accumulation of polysaccharides. Sci. Hortic. 240: 155-161.
Zhang, S., Y. Yang, J. Li, J. Qin, W. Zhang, W. Huang, H. Hu. 2018. Physiological diversity of orchids. Plant Divers. 40: 196-208.
Zheng, S. G., Y. D. Hu, R. X. Zhao, S. Yan, X. Q. Zhang, T. M. Zhao and Z. Chun. 2018. Genome-wide researches and applications on Dendrobium. Planta 248: 769-784.