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植物內生型抗蟲機制及抗蟲基因轉殖技術
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文心蘭開花生理的基因調控
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植物耐旱機制的基因選殖及基因表現研究
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ESTs library of Chinese cabbage roots colonized with Piriformospora indica
研究室簡介
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本實驗室是以甘藷為主要的實驗材料。因為甘藷具有病蟲害少、耐旱以及耐高溫等特性,這些都是極具發展性以及應用性的優點。因此,本實驗室一直以來便積極地自甘藷中尋找跟抗蟲、逆境相關而且具應用價值的基因。經過多年來的努力,我們找到了幾個重要的基因:
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抗蟲基因(SPORAMIN)-- SPORAMIN是甘藷塊根中含量非常豐富的儲存性蛋白質,其表現具有組織專一性,主要是在塊根中表現。經由實驗證明SPORAMIN具有胰蛋白酶抑制因子(trypsin inhibitor)的活性。之後,我們將SPORAMIN轉殖到菸草以及十字花科蔬菜當中,結果證實了這些轉殖株具有抵抗斜紋夜盜蛾幼蟲的啃食的能力(見圖一),目前我們正致力於利用SPORAMIN基因的MAR(matrix attachment region)來幫助轉殖的基因穩定並高度地在轉殖植物中表現,以避免基因靜默(gene silencing)的發生。除此之外,對於SPORAMIN基因啟動子的研究,我們也發現到SPORAMIN基因啟動子不但在轉殖菸草以及阿拉伯芥中具有受傷誘導的能力(見圖二),在轉殖馬鈴薯中也具有器官專一性表現的能力,主要是在塊莖中表現。這對我們在做植物基因轉殖上將會有很大的貢獻。
圖一
圖二
抗逆境基因(SPLTI)-- SPLTI是在甘藷未展開的幼葉中表現的蛋白質,在成熟的葉片中平常是不表現的,但是當甘藷受到乾旱或缺水的逆境時,在成熟的葉部則會表現。此外,SPLTI也是具有胰蛋白酶抑制因子(trypsin inhibitor)活性的蛋白質。另一方面,對於SPLTI基因啟動子的研究,我們經由序列的比對也發現到此啟動子上有許多跟植物逆境相關的順向活性因子(cis-acting element),如MYB、MYC以及DRE……等。經由實驗證明,在轉殖阿拉伯芥受到乾旱、低溫、高鹽度以及病菌感染的情況下,都有誘發此啟動子的作用。因此,目前我們正積極地研究轉殖SPLTI的阿拉伯芥對於抵抗逆境的表現情形。
幾年前,本實驗室也接下了一個文心蘭的國家型計畫。本實驗室主要負責的部分是研究文心蘭開花時,影響花序發育的基因。我們利用subtration以及cDNA microarray的方式,大量地篩選文心蘭花序發育前以及發育之後表現有差異性的基因,藉由此方法釣取出關鍵性的基因。目前我們已證實花序的發育和文心蘭假球莖中澱粉、葡萄醣跟蔗糖的含量有關,而且超氧化酶(peroxidase)可能扮演著關鍵性的角色。目前我們正在建構文心蘭的cDNA library,期望將超氧化酶的家族成員通通找出來,以利往後的研究。
目前實驗室中成員包括了:葉開溫教授(現為台大植科所所長)、一位博士後研究員、一位博士班學生、兩位研究助理、四位碩士班學生以及兩位大學部學生,共12名。我們誠摯地邀請您加入我們的研究團隊,一起為台灣的生技產業努力。
發表著作
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Meents, A. K., Chen, S. P., Reichelt, M., Lu, H. H., Bartram, S., Yeh, K. W.*, Mithofer, A. (2019) Volatile DMNT systemically induces jasmonate-independent direct anti-herbivore defense in leaves of sweet potato (Ipomoea batatas) plants. Sci. Rep. 9:17431.
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Chin, D. C., Senthilkumar R., Suen, C. S., Chien, C. Y., Hwang, M. J., Hsu, C. H., Xuhan, X., Lai, Z. X., Yeh, K. W. (2019) Plant Cytosolic Ascorbate Peroxidase with Dual Catalytic Activity Modulates Abiotic Stress Tolerances. iScience. 16: 31- 49.
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Ko, S. S., Liu, Y. C., Chung, M. C., Shih, M. C., Mohammadmehdi H., Oguchi T., Watanabe K. N., Yeh K. W. (2019) Environmental biosafety assessment on transgenic Oncidium orchid modified by RNA interference of Phytoene Synthase genes. Plant Biotechnol. 36: 181- 185.
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Liu, H., Senthilkumar, R,, Ma, G., Zou, Q., Zhu, K., Shen, X., Tian, D., Hua, M. S., Oelmüller, R., Yeh, K. W. (2019) Piriformospora indica-induced phytohormone changes and root colonization strategies are highly host-specific. Plant Signal Behav. 14 (9): 1632688.
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Lin, H. F., Xiong, J., Zhou, H. M., Chen, C. M., Lin, F. Z., Xu, X. M., Oelmüller, R., Xu, W. F., Yeh, K. W. (2019) Growth promotion and disease resistance induced in Anthurium colonized by the beneficial root endophyte Piriformospora indica. BMC Plant Biol. 19 (1) : 40.
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Liu, Y. C., Yeh, C. W., Chung, J. D., Tsai, C. Y., Chiou, C. Y., Yeh, K. W. (2019) Petal‐specific RNAi‐mediated silencing of the phytoene synthase gene reduces xanthophyll levels to generate new Oncidium orchid varieties with white‐colour blooms. Plant Biotechnol. J. 17 (11) : 2035- 2037.
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Chih-Wei Kao, Madhunita Bakshi,Irena Sherameti, Sheqin Dong, Michael Reichelt, Ralf Oelmuller and Kai-Wun Yeh*. (2016). A Chinese cabbage (Brassica campetris subsp. Chinensis) τ-type glutathione-S-transferase stimulates Arabidopsis development and primes against abiotic and biotic stress. Plant Mol Biol. [Epub ahead of print]
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Shi-Peng Chen, Chih-Hsien Kuo, Hsueh-Han Lu, Hui-Shan Lo and Kai-Wun Yeh*. (2016). The Sweet Potato NAC-Domain Transcription Factor IbNAC1 Is Dynamically Coordinated by the Activator IbbHLH3 and the Repressor IbbHLH4 to Reprogram the Defense Mechanism against Wounding. PLoS Genet. 12(10): e1006397
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Rajendran Senthil Kumar, Chin-Hui Shen, Pei-Yin Wu, Subbiah Suresh Kumar, Moda Sang Hua and Kai-Wun Yeh*. (2016). Nitric oxide participates in plant flowering repression by ascorbate. Scientific Reports. 6: 35246.
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Shi-Peng Chen, I Winnie Lin, Xuanyang Chen, Yin-Hao Huang, Shiao-Chi Chang, Hui-Shan Lo, Hseuh-Han Lu and Kai-Wun Yeh*. (2016). Sweet potato NAC transcription factor, IbNAC1, upregulates sporamin gene expression by binding the SWRE motif against mechanical wounding and herbivore attack.Plant J. 86(3): 234-48.
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Dan-Chu Chin, Chia-Chi Hsieh, Hsin-Yi Lin and Kai-Wun Yeh*. (2016). A Low Glutathione Redox State Couples with a Decreased Ascorbate Redox Ratio to Accelerate Flowering in Oncidium Orchid. Plant Cell Physiol.57(2): 423-36. doi: 10.1093/pcp/pcv206.
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Chii-Jeng Wang, Yuan-Li Chan, Chin Hui Shien and Kai-Wun Yeh*. (2015). Molecular characterization of fruit-specific class III peroxidase genes in tomato (Solanum lycopersicum). Journal of Plant Physiology. doi:10.1016/j.jplph.
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Jian-Xin Liu, Chung-Yi Chiou, Chin-Hui Shen, Peng-Jen Chen, Yao-Chung Liu, Chin-Der Jian,Xiao-Lan Shen, Fu-Quan Shen and Kai-Wun Yeh*. (2014). RNA interference-based gene silencing of phytoene synthase impairs growth, carotenoids, and plastid phenotype in Oncidium hybrid orchid. SpringerPlus. 3: 478
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Yuan-Li Chan, Yong He, Tsen-Tsz Hsiao, Chii-Jeng Wang, Zhihong Tian and Kai-Wun Yeh*. (2014). Pyramiding taro cystatin and fungal chitinase genes driven by a synthetic promoter enhances resistance in tomato to root-knot nematode Meloidogyne incognita. Plant Science. 231: 74-81.
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Chin DC, Shen CH, SenthilKumar R and Yeh KW*. (2014). Prolonged Exposure to Elevated Temperature Induces Floral Transition via Up-Regulation of Cytosolic Ascorbate Peroxidase 1 and Subsequent Reduction of the Ascorbate Redox Ratio in Oncidium Hybrid Orchid. Plant Cell Physiol. 55(12): 2164-76.
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SenthilKumar Rajendran, I-Winnie Lin, Mei-Ju Chen, Chien-Yu Chen and Kai-Wun Yeh*. (2014). Differential activation of sporamin expression in response to abiotic mechanical wounding and biotic herbivore attack in the sweet potato. BMC Plant Biology.14: 112
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Peng-Jen Chen, Rajendran Senthilkumar, Wann-Neng Jane, Yong He, Zhihong Tian and Kai-Wun Yeh.(2014).Transplastomic Nicotiana benthamiana plants expressing multiple defence genes encoding protease inhibitors and chitinase display broad-spectrum resistance against insects, pathogens and abiotic stresses. Plant Biotechnology Journal. DOI:10.1111/pbi.12157
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Wei Ye, Chin-Hui Shen, Yuling Lin, Peng-Jen Chen, Xuming Xu, Ralf Oelmuller, Kai-Wun Yeh, Zhongxiong Lai.(2014). Growth Promotion-Related miRNAs in Oncidium Orchid Roots Colonized by the Endophytic Fungus Piriformospora indica. PLoS One. 9(1): e84920.
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Dong SQ, Tian ZH, Chen PJ, Senthilkumar R, Shen CH, Cai D, Oelmullar R, Yeh KW.(2013).The maturation zone is an important target of Piriformospora indica in Chinese cabbage roots. Journal of Experimental Botany(doi:10.1093/jxb/ert265)
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Rajendran Senthilkumar and Kai-Wun Yeh.(2012). Multiple biological functions of sporamin related to stress tolerance in sweet potato (Ipomoea batatas Lam). Biotechnology Advances [Epub ahead of print]
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Xiao-Jing Liu, Yao-Nung Chuang, Chung-Yi Chiou, Dan-Chu Chin, Fu-Quan Shen, and Kai-Wun Yeh. (2012). Methylation effect on chalcone synthase gene expression determines anthocyanin pigmentation in floral tissues of two Oncidium orchid cultivars. Planta [Epub ahead of print]
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Ing-Feng Chang, Peng-Jen Chen, Chin-Hui Shen, Tsung-Ju Hsieh, Ya-Wen Hsu, Bau-Lian Huang, Ching-I Kuo, Yu-Ting Chen, Hsiu-An Chu, Kai-Wun Yeh, Li-Chun Huang (2010)Proteomic profiling of proteins associated with the rejuvenation of Sequoia sempervirens (D. Don) Endl. Proteome Science 8: 64 (doi: 10.1186/1477-).
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Yin-Chen Lee, Joy Michal Johnson, Ching-Te Chien, Chao Sun, Daguang Cai, Binggan Lou, Ralf Oelmuller, and Kai-Wun Yeh (2011) Growth Promotion of Chinese Cabbage and Arabidopsis by Piriformospora indica Is Not Stimulated by Mycelium-Synthesized Auxin. Molecular Plant-Microbe Interactions Vol. 24, No. 4, 2011, pp. 421–431.
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Y.-L. Chan, D. Cai, P. W. J. Taylor, M.-T. Chan and K.W. Yeh (2010) Adverse effect of the chitinolytic enzyme PjCHI-1 in transgenic tomato on egg mass production and embryonic development of Meloidogyne incognita. Plant Pathology Volume 59, Issue 5, pages 922–930
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Chung-Yi Chiou, Hsin-An Pan, Yao-Nung Chuang and Kai-Wun Yeh (2010) Differential expression of carotenoid-related genes determines diversified carotenoid coloration in floral tissues of Oncidium cultivars. Planta, 232:937-948 (SCI).
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Yuan-Li Chan, Ming-Tsair Chan, Daguang Cai, Kai-Wun Yeh (2010) A novel chitinolytic enzyme, PjCHI-1, causes aberrant embryonic development and egg mass production of Meloidogyne incognita in transgenic tomato. Plant pathology (in press)
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Yuan-Li Chan, Ai-Hwa Yang, Jen-Tzu Chen, Kai-Wun Yeh , Ming-Tsair Chan (2010) Heterologous expression of taro cystatin protects transgenic tomato against Meloidogyne incognita infection by means of interfering sex determination and suppressing gall formation. Plant Cell Reports 29:231-238
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Senthil Kumar, Cheng C.P. and Yeh K.W.(2010) Genetically pyramiding protease inhibitor genes for dual broad-spectrum resistance against insect and phytopathogens in transgenic tobacco. Plant Biotechnology Journal 8:65-75
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Chin-Hui Shen and Kai-Wun Yeh (2010) The signal network of ascorbate homeostasis. Plant Signaling & Behavior 5:1-3
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Shen C.H and Yeh K.W.(2010) Hydrogen peroxide mediates the expression of ascorbate-related genes in response to methanol stimulation in Oncidium. Journal of plant physiology 167(5):400-407
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Shen C. H., Krishnamurthy R. and Yeh K.W. (2009) Decreased L -Ascorbate Content Mediating Bolting is Mainly Regulated by the Galacturonate Pathway in Oncidium. Plant Cell Physiol. 50(5): 935-946 (SCI).
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Wang K.M., Rajendran S.K. Cheng Y.S., Shripathi V., Yang A.H. and Yeh K.W. (2008) Characterization of inhibitory mechanism and antifungal activity between group-1 and group-2 phytocystatin from taro (Colocasia esculenta). FEBS Journal 275: 4980-4989 (SCI).
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Chiou C.Y., Wu K.Q. and Yeh K.W. (2008) Characterization and promoter activity of chromoplast specific carotenoid associated gene (CHRC) from Oncidium Gower Ramsey. Biotechnology Letters 30: on-line (SCI).
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Ku A.T., Huang Y.S. Wang Y.S., Ma D. and Yeh K.W. (2008) IbMADS1 (Ipomoea batatas MADS-box 1 gene) is involved in tuberous root initiation in sweet potato. Annals of Botany 102: 57-67 (SCI).
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Chiou C.Y. and Yeh K.W. (2008) Differential expression of MYB gene (OgMYB1) determines color patterning in floral tissue of Oncidium Gower Ramsey. Plant Molecular Biology, 66:379-388 (SCI).
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Wang C.Y., Chiou C.Y., Wang H.L., Ramanarayan Krishnamurthy, Shripathi Venkatagiri, Tan J. and Yeh K.W. (2007) Carbohydrate mobilization and gene regulatory profile in the pseudobulb of Oncidium orchid during the flowering process. Planta, 227:1063-1077 (SCI).
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Chen C.C., Kumar H.G., Kumar S., Tzean S.S., Yeh K.W. (2007) Molecular cloning, characterization, and expression of a chitinase from the entomopathogenic fungus Paecilomyces javanicus. Curr. Microbiol. 55:8-13.
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Kumar, A., Ku, A.T. and Yeh, K.W. (2007) An efficient and rapid plant regeneration system for sweet potato [I. batatas(L.) Lam]. European J. Hort. Sci. 72(2) 85-89 (SCI)
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Chen, H. J., Wang, S. J., Chen. C. C. and Yeh, K.W. (2006) New gene construction strategy in T-DNA vector to enhance expression level of sweet potato sporamin and insect resistance in transgenic brassica oleracea. Plant Science 171:367-374
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A. H. Yang, and Yeh, K.W. (2005) Molecular cloning, recombinant gene expression,and anti (Colocasia esculenta cv. Kaosiung no. 1)fungal activity of cystatin from taro. Planta 221(4):493-501(SCI)
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A. H. Yang, and Yeh, K.W.(2005) Survey on gene expression of phytocystatin from local taro (Colocasia esculenta) plants. Crop, Environment & Bioinformatics,2:123-130
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Tan, J., Wang, H. L. and Yeh, K.W. (2005) Analysis of organ-specific, expressed genes in Oncidium orchid by subtractive expressed sequence tags library. Biotechnology Letters 27:1517-1528
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Wang, S. J., Yeh, K.W., and Tasi, C. Y. (2004) Circadian control of sweet potato granule-bound starch synthase I gene in Arabidopsis plants. Plant Growth Regulation 42:161-168(SCI)
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Cai, D., Lange, T., Tian, Y.Y., Thurau, T., Jung, C. and Yeh, K.W. (2003) Sporamin- mediated Resistance to Cyst Nematode (Heterodera schachtii Schm.) Is dependenet on Trypsin inhibitory Activity. Plant Molecular biology . Plant Mol. Biol.51:839-849 (SCI).
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Chen C.W. , Lei, B.C., Yeh K.W. and Duan K.J. (2003) Recombinant Sweet potato Sporamin Production via Glucose/pH Control in Fed-batch Cultures of Saccharomyces cerevisiae. Process Biochemistry 11:50-56(SCI).
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Wang, H.Y., Huang, Y.C., Chen, S.F. and Yeh, K.W. (2003) Molecular cloning, characterization and gene expression of a water deficiency and chilling induced proteinase inhibitor I gene family from sweet potato (Ipomoea batatas Lam.) leaves. Plant Science. 165:191-203(SCI)
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Y. C. Lan, S. F. Chen, S. J. Wang, Yeh, K. W. (2002) Wounding-responsive regulation of the promoter region of sporamin gene in sweet potato. Plant Mol. Biol. 48:223-231(SCI)
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Yao, P.L., Hwang, M. J.,Chen Y.M., Yeh, K. W. (2001) Site-directed mutagenesis evidence for a negatively charged trypsin inhibitory loop in sweet potato sporamin. FEBS letters. 496: 134-138 (SCI)
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Wang, C.K.,Duan, K.J., Yeh, K. W., Chen, W.C.(2001) Production of a fusion protein of sweet potato sporamin from recombinant E. coli XL1 Blue by fed-batch fermentations. Biotechnology Letters. 23:475-479(SCI).
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Wang, S.J., Yeh, K. W., Tsai, C.Y. (2001) Regulation of starch granule-bound starch synthase I gene expression by circadian clock and sucrose in the source tissue of sweet potato. Plant Science. 161:635-644 (SCI)
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Wang, S.J., Yeh, K. W., Tsai, C.Y. (1999) Molecular characterization and expression of a starch granule-bound starch synthase gene in the sink and source tissues of sweet potato. Physiologia Plantarum 106: 253-261(SCI).
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Ding, L.C., Hu, C.Y., Yeh, K.W. and Wang, P. J. (1998) Development of insect-resistant transgenic cauliflower plants expressing the trypsin inhibitor gene isolated from local sweet potato. Plant Cell Reports 17: 854-860 (SCI)
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Yu, Y.L., Yeh, K.W., and Lin, C.P. (1998) An antigenic protein genes of a phytoplasma associated with sweet potato witches' broom. Microbiology 144: 1257-1262(SCI)
開設課程
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分子生物學
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植物訊息傳導學
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基因定點突變學
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植物功能性基因組學及實驗法