转基因抗烟嘧磺隆水稻的研究
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摘要
烟嘧磺隆(Nicosulfuron)是一种超高效、广谱的磺酰脲类除草剂,大量用于玉米田苗后的杂草防治。对于玉米田中发生的多数一年生禾本科杂草、部分阔叶杂草及部分莎草科杂草具有较好的防治效果。烟嘧磺隆易被植物的根、叶吸收,在木质部和韧皮部传导,其作用靶标是乙酰乳酸合成酶(acetolactate synthetase, ALS),而ALS是植物合成支链氨基酸撷氨酸、异亮氨酸、亮氨酸的一个关键酶,由于ALS的活性严重受到抑制,导致缬氨酸、异亮氨酸、亮氨酸缺乏,影响植物细胞的有丝分裂,从而导致植物死亡。我国于20世纪90年代初期开始推广烟嘧磺隆,由于其除草效果突出、可混性强、用量低等优点而广受欢迎。
本论文通过生物信息学技术,从玉米自交系B73基因组中选择了5个可能参与烟嘧磺隆降解的基因。其中一个基因导入到水稻以后使水稻获得了抗烟嘧磺隆的能力。这个基因被命名为253。转基因抗烟嘧磺隆水稻的抗性通过在大田六块田间区域内,以40mL/m2的使用量分别喷施80mg/L、160mg/L、320mg/L、480mg/L、640mg/L浓度的烟嘧磺隆和清水对照获得。结果表明253基因水稻植株在正常的玉米田内烟嘧磺隆除草剂使用浓度(80mg/L)下生长不受影响,部分转基因株系对烟嘧磺隆的抗性水平甚至可以达到640mg/L,大大超过了田间杂草防治的使用浓度。
本研究获得的转基因水稻可以在苗后利用烟嘧磺隆防治杂草。烟嘧磺隆杀草效果好、成本低,因此本研究获得的转基因水稻具有重要应用价值。
Nicosulfuron is sulfonylurea herbicide. It is an ultra-efficient, broad-spectrum postemergence corn herbicide, and widely used for control of annual grass weeds, broadleaf weeds and some Cyperaceae in Maize fields. Nicosulfuron can be easily absorbed in plant roots, leaves, and is transported in the xylem and phloem. Nicosulfuron is an inhibitor of acetolactare synthase(ALS), which is a key Enzyme for the biosynthesis of all the branched-chain amino acid, valine, isoleucine, and leucine. Nicosulfuron is a widely used herbicide since the early 90's.
Five possible P450 genes were selected by bioinformatic analysis of the genomic sequences of the maize inbred line B73. All the 5 genes were transformed into rice by Agribacterium-medicated transformation. The transformed rice events from one gene, name 253, were found to be tolerant to nicosulfuron. From field trials of nicosulfuron, showed that all 253 transgenic rice events can grow well within the normal nicrosulfuron herbicide use concentration(80mg/L) of com, part of the transgenic rice events of nicosulfuron resistance level can reach 640mg/L.
This study created nicosulfuron tolerant transgenic rice. We believe these transgenic rice could be utilized for weed control mangerment for rice field.
本论文通过生物信息学技术,从玉米自交系B73基因组中选择了5个可能参与烟嘧磺隆降解的基因。其中一个基因导入到水稻以后使水稻获得了抗烟嘧磺隆的能力。这个基因被命名为253。转基因抗烟嘧磺隆水稻的抗性通过在大田六块田间区域内,以40mL/m2的使用量分别喷施80mg/L、160mg/L、320mg/L、480mg/L、640mg/L浓度的烟嘧磺隆和清水对照获得。结果表明253基因水稻植株在正常的玉米田内烟嘧磺隆除草剂使用浓度(80mg/L)下生长不受影响,部分转基因株系对烟嘧磺隆的抗性水平甚至可以达到640mg/L,大大超过了田间杂草防治的使用浓度。
本研究获得的转基因水稻可以在苗后利用烟嘧磺隆防治杂草。烟嘧磺隆杀草效果好、成本低,因此本研究获得的转基因水稻具有重要应用价值。
Nicosulfuron is sulfonylurea herbicide. It is an ultra-efficient, broad-spectrum postemergence corn herbicide, and widely used for control of annual grass weeds, broadleaf weeds and some Cyperaceae in Maize fields. Nicosulfuron can be easily absorbed in plant roots, leaves, and is transported in the xylem and phloem. Nicosulfuron is an inhibitor of acetolactare synthase(ALS), which is a key Enzyme for the biosynthesis of all the branched-chain amino acid, valine, isoleucine, and leucine. Nicosulfuron is a widely used herbicide since the early 90's.
Five possible P450 genes were selected by bioinformatic analysis of the genomic sequences of the maize inbred line B73. All the 5 genes were transformed into rice by Agribacterium-medicated transformation. The transformed rice events from one gene, name 253, were found to be tolerant to nicosulfuron. From field trials of nicosulfuron, showed that all 253 transgenic rice events can grow well within the normal nicrosulfuron herbicide use concentration(80mg/L) of com, part of the transgenic rice events of nicosulfuron resistance level can reach 640mg/L.
This study created nicosulfuron tolerant transgenic rice. We believe these transgenic rice could be utilized for weed control mangerment for rice field.
引文
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Cao M X, Huang J Q, Wei Z M, Yao Q H, Wan C Z and Lu J A. Engineering higher yield and herbicide resistance in rice by A grobacterium-mediated multiple gene transformation. Crop Sci.2004,44(6):2206-2213
Diehl K E and E W Stoller. Effect of Simulated Rainfall, Insecticide Formulation, and Insecticide Application Method on the Interaction between Nicosulfuron and Terbufosin Corn (Zea mays). Weed Tech.1995,9:80-85
Devine, Malcolm D. Mechanisms of resistance to acetyl-coenzyme a carboxylase inhibitors. Pesticide Science.1998,51:259-264
Danidle Werch-Reichhart, Alain Hehn and Luc Didierjean. Cytochromes P450 for engineering herbicide tolerance. Trends in plant science.2000,5(3):116-123
Duggleby R G, Pang S S. Acetohydroxyacid Synthase. JBiochem Mol Biol.2000,33: 1-36
Deng L H, Yu Y J, Li B J, Xiao G Y. Construction on plant expression vector containing multiple insect-resistant genes with Epsps as selection marker. Biotechnology.2008,18(1):9-13
Green J M and J F Ulrich. Response of Corn (Zea mays L.) Inbreds and Hybrids to Sulfonylurea Herbicides. Weed Sci.1993,41:508-516
Gnozalez J M and L Ukrainczyk. Adsorption and Desorption of Nicosulfuron in Soils. J. Environ. Qual.1996,25:1186-1192
Gaston S, Ribas-Carbo M, Busquets S. Changes in mitochondrial electron partitioning in response to herbcides inhibiting branched-chain amino acid biosynthesis in soybean. Plant Physiology.2003,133:1351-1359
Gang Pan. Xianyin Zhang, Kede Liu, Jiwen Zhang, Xiaozhi Wu, Jun Zhu, and Jumin Tu. Map-based cloning of a novel rice cyto chrome P450 gene CYP81A6 that confers resistance to two different classes of herbicides. Plant Molecular Biology. 2006,61(6):933-943
Hill C M, Duggleby R G. Mutagenesis of Escherichia coli acetohydroxy acid synthase 2 and characterization of three herbicide-resistant forms. Biochem J.1998,335: 653-661
Hideo Ohkawa, Hisae Tsujii, Yasunobu Ohkawa. The use of cytochrome P450 genes to introduce herbicide tolerance in crops:a review. Pesticide Science.1999,55: 867-874
Heap I. The international survey of herbicide resistant weeds [EB/OL]. [2009-12-20]. http://www.weedscience.com.
James V, Hay. Chemistry of sulfonylurea herbicides. Pesticide Science.1990,3 (29): 247-261
Jun Y, Songnian H, Jian W, et al. A draft sequence of the rice genome (Oryza saliva L. ssp indica). Science.2002,296:79-92
Kling J. Could transgenic super crops one day breed super weeds? Science.1996, 274:180-181
Kawahigashi H, Hirose S, Inui H, Ohkawa H and Ohkawa Y. Enhanced herbicide cross-tolerance in transgenic rice plants co-expressing human CYP1A1, CYP2B6, and CYP2C19, Plant Sci.2005,168(3):773-781
Lou zhongxun, wu Ray. A simple method for the transformation of rice is via the pollen-tube pathway. Plant molecular Reporter.1998,6(3):165-174
Lu C G, Zhou J S. Breeding and utilization of two-line inter-subspecific hybrid rice Liangyoupcijiu. Hybrid Rice.2000,15(2):4-5.
Lee S H, Park H J, Cho S Y, Chun H K, Park Y H, Jeong M H and Park S H. Evaluation of nutritional safety for the herbivide-resistant rice in growing male rats. Korean J. Nutr.2003,36(10):1030-1035
Lee Y, Jung S and Back K. Expression of human protoporphyrinogen oxidase in transgenic rice induces both a photodynamic response and oxyfluorfen resistance. Pest Biochem Physiol.2004,80(2):65-74
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