Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/23067
標題: 同時表現藍綠菌ictB基因與水稻OsCKX2 small interference RNA於台農67號以提升產量
Simultaneous expression of cyanobacterial ictB gene and rice OsCKX2 small interference RNA in TNG67 to improve its productivity
作者: 葉上寧
Yeh, Shang-Ning
關鍵字: TNG67
台農67號
ictB
CX5
productivity
ictB
CX5
產量
出版社: 生命科學系所
引用: Ashikari, M., Sakakibara, H., Lin, S., Yamamoto, T., Takashi, T., Nishimura, A., Angeles, E.R., Qian, Q., Kitano, H., and Matsuoka, M. (2005). Cytokinin oxidase regulates rice grain production. Science 309, 741-745. Berner, R.A. (1990). Atmospheric carbon dioxide levels over phanerozoic time. Science 249, 1382-1386. Bonfil, D.J., Ronen-Tarazi, M., Sultemeyer, D., Lieman-Hurwitz, J., Schatz, D., and Kaplan, A. (1998). A putative HCO3- transporter in the cyanobacterium Synechococcus sp. strain PCC 7942. FEBS Lett. 430, 236-240. Buick, R. (1992). The antiquity of oxygenic photosynthesis: evidence from stromatolites in sulphate-deficient Archaean lakes. Science 255, 74-77. FAO. (2009). How to Feed the World in 2050. available at http://www.fao.org/wsfs/forum2050/wsfs-background-documents/hlef-issues-briefs/en/. Galuszka, P., Frebortova, J., Werner, T., Yamada, M., Strnad, M., Schmulling, T., and Frebort, I. (2004). Cytokinin oxidase/dehydrogenase genes in barley and wheat. European Journal of Biochemistry 271, 3990-4002. Lieman-Hurwitz, J., Rachmilevitch, S., Mittler, R., Marcus, Y., and Kaplan, A. (2003). Enhanced photosynthesis and growth of transgenic plants that express ictB, a gene involved in HCO3- accumulation in cyanobacteria. Plant Biotechnol. J. 1, 43-50. McGaw, B.A., and Horgan, R. (1983). Cytokinin catabolism and cytokinin oxidase. Phytochemistry 22, 1103-1105. Omata, T., Price, G.D., Badger, M.R., Okamura, M., Gohta, S., and Ogawa, T. (1999). Identification of an ATP-binding cassette transporter involved in bicarbonate uptake in the cyanobacterium Synechococcus sp. strain PCC 7942. PNAS of the U.S.A. 96, 13571-13576. Pearson, P.N., and Palmer, M.R. (2000). Atmospheric carbon dioxide concentrations over the past 60 million years. Nature 406, 695-699. Price, G.D., Badger, M.R., Woodger, F.J., and Long, B.M. (2008). Advances in understanding the cyanobacterial CO2-concentrating-mechanism (CCM): functional components, Ci transporters, diversity, genetic regulation and prospects for engineering into plants. J. Exp. Bot. 59, 1441-1461. Price, G.D., Woodger, F.J., Badger, M.R., Howitt, S.M., and Tucker, L. (2004). Identification of a SulP-type bicarbonate transporter in marine cyanobacteria. PNAS of the U.S.A. 101, 18228-18233. Schmulling, T., Werner, T., Riefler, M., Krupkova, E., and Bartrina y Manns, I. (2003). Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species. Journal of Plant Research 116, 241-252. Shibata, M., Katoh, H., Sonoda, M., Ohkawa, H., Shimoyama, M., Fukuzawa, H., Kaplan, A., and Ogawa, T. (2002). Genes essential to sodium-dependent bicarbonate transport in cyanobacteria. Journal of Biological Chemistry 277, 18658-18664. Werner, T., Motyka, V., Laucou, V., Smets, R., Van Onckelen, H., and Schmulling, T. (2003). Cytokinin-deficient transgenic arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity. Plant Cell 15, 2532-2550. Zhang, Q. (2007). Strategies for developing green super rice. PNAS of the U.S.A. 104, 16402-16409.
摘要: 即使科技不斷進步,世界上依然存在糧食缺乏的問題,且隨著人口的膨脹以及生質能源的發展,未來糧食的缺口將逐漸擴大,為了在有限的土地面積上提供更多糧食,因此各國皆致力於高產作物的育種。本試驗透過基因轉殖,在水稻上同時表現藍綠菌中參與碳酸根運輸的相關蛋白基因ictB與水稻內生OsCKX2 的外源干擾RNA (interference RNA)基因CKX5,一方面提高轉殖株的固碳能力,以累積較多的生物量(biomass);另一方面抑制細胞分裂素氧化酶(cytokinin oxidase)的基因表現,以提升細胞分裂素(cytokinin)含量,進而促進水稻的分化與發育,並期望綜合兩個基因之特性以增加水稻作物產量。 首先從篩選T1轉殖株的外源基因開始,經由報導蛋白GUS染色面積的比較與種子在hygromycin篩選中的發芽比例,取得可能含有完整T-DNA的七個T2轉殖品系(Line 3-2-1、3-2-4、5-1-18、5-1-20、6-2-1、6-2-10和7-1-5),作為後續檢測的材料。經過專一性引子的PCR檢測,以及專一性探針進行ictB和CKX5基因的偵測後,再一次以葉片GUS染色和種子hygromycin發芽試驗,篩選出含有完整T-DNA的T2轉殖株。經南方墨點法檢測結果,顯示品系3-2-1-7、3-2-4-1和3-2-4-2含單套T-DNA,品系6-2-1-7、6-2-10-1和7-1-5-5含雙套T-DNA。進一步以TAIL-PCR找出T-DNA的插入位置,並由後續的PCR結果中顯示,品系3-2-1-7、3-2-4-1、3-2-4-2、6-2-10-1和7-1-5-5為同質合子,6-2-1-7則為異型合子;但在含有hygromycin的培養基中6-2-1-7仍有較高的發芽率,因此推測其另一套T-DNA亦可能為同質合子。綜合以上的結果顯示本研究已從轉殖株中,篩選到單套同質合子,而含雙套T-DNA的轉殖株只能確定至少有一套為同質合子,且位於T-DNA兩端的GUS和hptII都能正常表現,故推測含於T-DNA中段的ictB和CKX5也應可表現。 在生長特性上,初步的田間試驗結果發現,T2品系3-2-4-1、3-2-4-6、6-2-1-10以及7-1-5-4的每株穗數(有效分蘗數)與種子粒數都較台農67號有顯著的提升,尤其6-2-10-1提升超過100%最為可觀,此外該品系上在地表總乾物重和實粒重極顯著的增加將近一倍;7-1-5-4也得到類似的結果,唯差異較不明顯,而品系 3-2-4-1和3-2-4-6則只有在總粒數與穗數上有明顯的提高,顯示轉殖株在生長量上有較佳的表現,且可增加單株水稻的實粒重,由此可證實ictB與CKX5的同時表現的確可以提高水稻產量。
URI: http://hdl.handle.net/11455/23067
其他識別: U0005-1708201016513700
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1708201016513700
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