菊花抗白色锈病相关基因CmDREBa-2的克隆及表达分析
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摘要
以菊花(Chrysanthemum morifolium)免疫白色锈病品种‘C029’为试验材料,基于白色锈病病原菌诱导的菊花转录组数据库,采用RACE(rapid-amplification of c DNA ends)方法克隆得到菊花DREB(Dehydration responsive element binding protein)基因,命名为CmDREBa-2。对其进行生物信息学分析和表达分析,结果表明:CmDREBa-2开放阅读框(ORF)全长291 bp,编码96个氨基酸;预测CmDREBa-2蛋白相对分子质量为11 159.04,p I为11.34,是一个定位于细胞核、亲水的不稳定蛋白;氨基酸序列和结构分析显示该蛋白含有1个由49个氨基酸残基组成的AP2保守结构域,所以该蛋白属于AP2/EREBP家族;进化树分析表明Cm DREBa-2属于DREB家族的A-1组,CmDREBa-2蛋白与菊花的其他两个DREB类蛋白Cm DREBa和CmDREBb的同源性较高;菊花‘C029’接种白色锈病病原菌6 h后,CmDREBa-2表达量达到最高,约为未接菌对照的34倍;用水杨酸(SA)、茉莉酸甲酯(Me JA)和乙烯利(ETH)处理‘C029’菊花叶片,结果表明CmDREBa-2受3种激素的诱导表达。
Based on the information of transcriptome database derived from‘C029',a Puccinia horiana Henn. immune Chrysanthemum morifolium cultivar,the c DNA sequence of DREB(Dehydration responsive element binding protein)was cloned. This DREB gene was named CmDREBa-2. Sequence analysis showed that the open reading frame(ORF)of CmDREBa-2 was 291 bp,encoding 96 amino acids. The molecular weight of the predicted protein was 11 159.04 and the p I value was 11.34. The protein was located in the nucleus,and it was a hydrophilic and unstable protein. The transcription factor of CmDREBa-2 contained the AP2 DNA binding domain which was composed by 49 amino acids. The results of phylogenetic analysis showed that the CmDREBa-2 belonged to the DREB family of A-1 and it had a close relationship with CmDREBa and Cm DREBb from C. morifolium. The expression level of CmDREBa-2 reached a maximum at 6 h after Puccinia horiana Henn. infection,with 34 times as much as the control. We also found that the expression of CmDREBa-2 was induced by SA,Me JA and ETH.
Based on the information of transcriptome database derived from‘C029',a Puccinia horiana Henn. immune Chrysanthemum morifolium cultivar,the c DNA sequence of DREB(Dehydration responsive element binding protein)was cloned. This DREB gene was named CmDREBa-2. Sequence analysis showed that the open reading frame(ORF)of CmDREBa-2 was 291 bp,encoding 96 amino acids. The molecular weight of the predicted protein was 11 159.04 and the p I value was 11.34. The protein was located in the nucleus,and it was a hydrophilic and unstable protein. The transcription factor of CmDREBa-2 contained the AP2 DNA binding domain which was composed by 49 amino acids. The results of phylogenetic analysis showed that the CmDREBa-2 belonged to the DREB family of A-1 and it had a close relationship with CmDREBa and Cm DREBb from C. morifolium. The expression level of CmDREBa-2 reached a maximum at 6 h after Puccinia horiana Henn. infection,with 34 times as much as the control. We also found that the expression of CmDREBa-2 was induced by SA,Me JA and ETH.
引文
Bonde M R,Murphy C A,Bauchan G R,Luster D G,Palmer C L,Nester S E.2015.Evidence for systemic infection by Puccinia horiana,causal agent of chrysanthemum white rust,in chrysanthemum.Phytopathology,105(1):91-98.
Bonde M R,Palmer C L,Luster D G,Nester S E,Revell J M,Berger D K.2014.Viability of Puccinia horiana teliospores under various environmental conditions.Plant Health Progress,15(1):25-28.
Chen J Q,Dong Y,Wang Y J,Liu Q,Zhang J S,Chen S Y.2003.An AP2/EREBP-type transcription-factor gene from rice is cold-inducible and encodes a nuclear-localized protein.Theoretical and Applied Genetics,107(6):972-979.
Chen Y X,Huang L K,Yan H D,Zhang X Q,Xu B,Ma X.2016.Cloning and characterization of an ABA-independent DREB transcription factor gene,HcD REB2,in Hemarthria compressa.Hereditas,153(1):3.
Chini A,Grant J J,Seki M,Shinozaki K,Loake G J.2004.Drought tolerance established by enhanced expression of the CC-NBS-LRR gene,ADR1,requires salicylic acid,EDS1 and ABI1.Plant Journal for Cell&Molecular Biology,38(5):810-822.
Ding Shi-min,Xi Dun-qin.2001.Occurrence regularity and chemical control of chrysanthemum white rust.Plant Protection,27(2):20-22.(in Chinese)丁世民,席敦芹.2001.菊花白锈病发生规律与药剂防治.植物保护,27(2):20-22.
Dong L,Huang Z Q,Liu D,Zhu P F,LüS J,Li N,Mao H Y.2018.Transcriptome analysis of chrysanthemum in responses to white rust.Scientia Horticulturae,233:421-430.
Dubouzet J G,Sakuma Y,Ito Y,Kasuga M,Dubouzet E G.2003.Os DREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought-high-salt-and cold-responsive gene expression.Plant Journal for Cell&Molecular Biology,33(4):751-763.
Fan Wen-zhong,Wei Guo-xian,Sun Yan-mei.2002.Chrysanthemum white rust occurred seriously in Jilin.Plant Protection,28(2):20.(in Chinese)范文忠,魏国先,孙艳梅.2002.菊花白锈病在吉林市严重发生.植物保护,28(2):20.
Feng Jian-xun,Liu Di,Pan Yi,Gong Wei,Ma Li-geng,Luo Jing-chu,Deng Xing-wang,Zhu Yu-xian.2005.An annotation update via cD NAsequence analysis and comprehensive profiling of developmental,hormonal or environmental responsiveness of the Arabidopsis AP2/EREBPtranscription factor gene family.Plant Molecular Biology,59:853-868.
Herath V.2016.Small family,big impact:in silico analysis of DREB2 transcription factor family in rice.Computational Biology&Chemistry,65:128-139.
Hong Bo,Tong Zheng,Ma Nan,Li Jian-ke,Gao Jun-ping.2006.The heterologous expression of At DREB1A gene in Chrysanthemum morifolium enhanced the tolerance of plants to drought and salinity stress.Chinese Science C series(Life Sciences),36(3):223-231.(in Chinese)洪波,仝征,马男,李建科,高俊平.2006.At DREB1A基因在菊花中的异源表达提高了植株对干旱和盐渍胁迫的耐性.中国科学C辑(生命科学),36(3):223-231.
Ji Ai-jia,Luo Hong-mei,Xu Zhi-chao,Zhang Xin,Song Jing-yuan,Chen Shi-lin.2015.Research and perspectives on AP2/ERF transcription factors in medicinal plants.Chinese Science Bulletin,(14):1272-1284.(in Chinese)季爱加,罗红梅,徐志超,张鑫,宋经元,陈士林.2015.药用植物转录因子AP2/ERF研究与展望.科学通报,(14):1272-1284.
Lai Y,Dang F,Lin J,Yu L,Shi Y,Xiao Y,Huang M,Lin J,Chen C,Qi A,Liu Z,Guan D,Mou S,Qiu A,He S.2013.Overexpression of a Chinese cabbage Br ERF11 transcription factor enhances disease resistance to Ralstonia solanacearum in tobacco.Plant Physiology Biochemistry,62(1):70-78.
Li Yong-peng,Zhang Jia-jia,Zhang Li-wei,Tan Ji,Chen Meng-ya,Du Li.2016.Isolation of two DREB1 transcription factor genes and their responses to different stresses in Toona sinensis.Acta Horticulturae Sinica,43(4):743-751.(in Chinese)李勇鹏,张佳佳,张力维,谭计,陈梦雅,杜丽.2016.香樟两个DREB1转录因子基因的分离及其对不同胁迫的响应分析.园艺学报,43(4):743-751.
Liang Chun-bo,Huang Xu-tang,Li Cen,Zhou Fei,Wang Wen-jun,Ma Jun,Chen Hui-rong,Guo Yong-li.2015.Cloning and expressing analysis of a DREB transcription factor in‘Sunflower’.Ningxia Journal of Agriculture and Forestry Science and Technology,56(7):31-34.(in Chinese)梁春波,黄绪堂,李岑,周菲,王文军,马军,陈惠荣,郭永利.2015.向日葵逆境应答转录因子DREB的克隆与表达分析.宁夏农林科技,56(7):31-34.
Liang Wei.2003.Investigation results of rust resistance of introduced chrysanthemum varieties in Lanzhou area and measures to prevent rust.Gansu Agricultural Science and Technology,(9):41-43.(in Chinese)梁伟.2003.兰州地区引进菊花品种的抗锈性调查结果及防锈病措施.甘肃农业科技,(9):41-43.
Licausi F,Ohme-Takagi M,Perata P.2013.APETALA2/ethylene responsive factor(AP2/ERF)transcription factors:mediators of stress responses and developmental programs.New Phytologist,199(3):639-649.
Liu L Q,Zhu K,Yang Y F,Wu J,Chen F D,Yu D Y.2008.Molecular cloning,expression profiling and trans-activationproperty studies of a DREB2-like gene from chrysanthemum(Dendranthema vestitum).Journal of Plant Research,121(2):215-226.
Liu Xiao-ying,Chen Li-yuan,Zhang Jing-qiu,Li Jia-wei,Gao Yue,Wang Zhen-ying.2013.Isolation and functional analysis of a new DREBtranscription factor(Bp DREB1)from Brassica pekinensis.Acta Agronomica Sinica,39(2):230-237.(in Chinese)刘晓颖,陈丽媛,张竞秋,李嘉玮,高越,王振英.2013.白菜脱水应答转录因子Bp DREB1基因的克隆及功能研究.作物学报,39(2):230-237.
Liu Zhi-wei,Xiong Yang-yang,Li Tong,Yan Ya-jun,Han Hong-run,Wu Zhi-jun,Zhuang Jing.2015.The cloning of transcription factor gene Cs DREB-A4 and the response to temperature stress in Camellia sinensis.Journal of Tea Science,35(1):24-34.(in Chinese)刘志薇,熊洋洋,李彤,严雅君,韩洪润,吴致君,庄静.2015.茶树转录子基因Cs DREB-A4的克隆与温度胁迫响应的分析.茶叶科学,35(1):24-34.
Lu J,Robert C A,Riemann M,Cosme M,Mène-SaffranéL,Massana J,Stout M J,Lou Y,Gershenzon J,Erb M.2015.Induced jasmonate signaling leads to contrasting effects on root damage and herbivore performance.Plant Physiology,167(3):1100-1116.
Nakano T,Suzuki K,Ohtsuki N,Tsujimoto Y,Fujimura T,Shinshi H.2006.Identification of genes of the plant-specific transcription-factor families cooperatively regulated by ethylene and jasmonate in Arabidopsis thaliana.Journal of Plant Research,119(4):407-413.
Pernas M,Sánchez-Monge R,Salcedo G.2000.Biotic and abioticstresses can induce cystatin expression in chestnut.Febs Letters,467(2-3):206-210.
Sakuma Y,Liu Q,Dubouzet J G,Abe H,Shinozaki K,Yamaguchi-Shinozaki K.2002.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression.Biochemical and Biophysical Research Communications,290:998-1009.
Sriram S,Chandran N K,Kumar R,Reddy M K.2015.First report of Puccinia horiana causing white rust of chrysanthemum in India.New Disease Reports,32:8.
Stockinger E J,Gilmour S J,Thomashow M F.1997.Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE,a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit.Proceedings of the National Academy of Sciences,94(3):1035-1040.
Tian Xiu-ling,Xia Bing-qiang,Luo feng-xia,Deng li-qin.1999.A preliminary study on the pathogen and infection pathway of white rust in Chrysanthemum.Journal of Shenyang Aricultural University,30(3):379-380.(in Chinese)田秀玲,夏炳强,罗凤霞,邓丽琴.1999.菊花白色锈病的病原菌和侵染途径的初步研究.沈阳农业大学学报,30(3):379-380.
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Xie Deng-lei,Cui Jiang-hui,Chang Jin-hua.2013.Cloning and expression analysis of Sb DREB2 gene from Sorghum bicolor.Acta Agronomica Sinica,39(8):1352-1359.(in Chinese)谢登雷,崔江慧,常金华.2013.高粱中Sb DREB2基因的克隆与表达分析作.作物学报,39(8):1352-1359.
Yang Ying-jie,Li Chun-shui,Zhang Xiao-jiao,Wei Qian,Gao Jun-ping,Hong Bo.2013.Drought stress tolerance of the hybrid offspring produced by At DREB1A transgenic chrysanthemum.Journal of Agricultural Biotechnology,21(2):148-157.(in Chinese)杨英杰,李春水,张晓娇,魏倩,高俊平,洪波.2013.转At DREB1A基因菊花杂交后代优株水分胁迫耐性分析.农业生物技术学报,21(2):148-157.
Yao W J,Fu Y R,Zhang Y F,Li H.2016.Cloning of four DREB genes from Tibetan Sophora moorcroftiana and analysis of their expression during abiotic stress.Journal of Forestry Research,27(3):675-683.
Yu Yang,Wu Ping,Li Chun-shui,Hong Bo.2011.Drought tolerance and water saving ability in DREB1A transgenic plants of ground cover chrysanthemum‘Fall Color’.Journal of Northeast Forestry University,39(8):33-39.(in Chinese)于洋,吴萍,李春水,洪波.2011.转DREB1A基因地被菊耐旱节水性.东北林业大学学报,39(8):33-39.
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Zhang Yong,Tang hao-ru,Luo Ya,Wang Xiao-rong,Chen Qing,Liu Ze-jing.2014.Cloning and expression analysis of strawberry Fa CBF1 gene.Acta Horticulturae Sinica,41(2):240-248.(in Chinese)张勇,汤浩茹,罗娅,王小蓉,陈清,刘泽静.2014.草莓Fa CBF1基因的克隆及表达分析.园艺学报,41(2):240-248.
Bonde M R,Palmer C L,Luster D G,Nester S E,Revell J M,Berger D K.2014.Viability of Puccinia horiana teliospores under various environmental conditions.Plant Health Progress,15(1):25-28.
Chen J Q,Dong Y,Wang Y J,Liu Q,Zhang J S,Chen S Y.2003.An AP2/EREBP-type transcription-factor gene from rice is cold-inducible and encodes a nuclear-localized protein.Theoretical and Applied Genetics,107(6):972-979.
Chen Y X,Huang L K,Yan H D,Zhang X Q,Xu B,Ma X.2016.Cloning and characterization of an ABA-independent DREB transcription factor gene,HcD REB2,in Hemarthria compressa.Hereditas,153(1):3.
Chini A,Grant J J,Seki M,Shinozaki K,Loake G J.2004.Drought tolerance established by enhanced expression of the CC-NBS-LRR gene,ADR1,requires salicylic acid,EDS1 and ABI1.Plant Journal for Cell&Molecular Biology,38(5):810-822.
Ding Shi-min,Xi Dun-qin.2001.Occurrence regularity and chemical control of chrysanthemum white rust.Plant Protection,27(2):20-22.(in Chinese)丁世民,席敦芹.2001.菊花白锈病发生规律与药剂防治.植物保护,27(2):20-22.
Dong L,Huang Z Q,Liu D,Zhu P F,LüS J,Li N,Mao H Y.2018.Transcriptome analysis of chrysanthemum in responses to white rust.Scientia Horticulturae,233:421-430.
Dubouzet J G,Sakuma Y,Ito Y,Kasuga M,Dubouzet E G.2003.Os DREB genes in rice,Oryza sativa L.,encode transcription activators that function in drought-high-salt-and cold-responsive gene expression.Plant Journal for Cell&Molecular Biology,33(4):751-763.
Fan Wen-zhong,Wei Guo-xian,Sun Yan-mei.2002.Chrysanthemum white rust occurred seriously in Jilin.Plant Protection,28(2):20.(in Chinese)范文忠,魏国先,孙艳梅.2002.菊花白锈病在吉林市严重发生.植物保护,28(2):20.
Feng Jian-xun,Liu Di,Pan Yi,Gong Wei,Ma Li-geng,Luo Jing-chu,Deng Xing-wang,Zhu Yu-xian.2005.An annotation update via cD NAsequence analysis and comprehensive profiling of developmental,hormonal or environmental responsiveness of the Arabidopsis AP2/EREBPtranscription factor gene family.Plant Molecular Biology,59:853-868.
Herath V.2016.Small family,big impact:in silico analysis of DREB2 transcription factor family in rice.Computational Biology&Chemistry,65:128-139.
Hong Bo,Tong Zheng,Ma Nan,Li Jian-ke,Gao Jun-ping.2006.The heterologous expression of At DREB1A gene in Chrysanthemum morifolium enhanced the tolerance of plants to drought and salinity stress.Chinese Science C series(Life Sciences),36(3):223-231.(in Chinese)洪波,仝征,马男,李建科,高俊平.2006.At DREB1A基因在菊花中的异源表达提高了植株对干旱和盐渍胁迫的耐性.中国科学C辑(生命科学),36(3):223-231.
Ji Ai-jia,Luo Hong-mei,Xu Zhi-chao,Zhang Xin,Song Jing-yuan,Chen Shi-lin.2015.Research and perspectives on AP2/ERF transcription factors in medicinal plants.Chinese Science Bulletin,(14):1272-1284.(in Chinese)季爱加,罗红梅,徐志超,张鑫,宋经元,陈士林.2015.药用植物转录因子AP2/ERF研究与展望.科学通报,(14):1272-1284.
Lai Y,Dang F,Lin J,Yu L,Shi Y,Xiao Y,Huang M,Lin J,Chen C,Qi A,Liu Z,Guan D,Mou S,Qiu A,He S.2013.Overexpression of a Chinese cabbage Br ERF11 transcription factor enhances disease resistance to Ralstonia solanacearum in tobacco.Plant Physiology Biochemistry,62(1):70-78.
Li Yong-peng,Zhang Jia-jia,Zhang Li-wei,Tan Ji,Chen Meng-ya,Du Li.2016.Isolation of two DREB1 transcription factor genes and their responses to different stresses in Toona sinensis.Acta Horticulturae Sinica,43(4):743-751.(in Chinese)李勇鹏,张佳佳,张力维,谭计,陈梦雅,杜丽.2016.香樟两个DREB1转录因子基因的分离及其对不同胁迫的响应分析.园艺学报,43(4):743-751.
Liang Chun-bo,Huang Xu-tang,Li Cen,Zhou Fei,Wang Wen-jun,Ma Jun,Chen Hui-rong,Guo Yong-li.2015.Cloning and expressing analysis of a DREB transcription factor in‘Sunflower’.Ningxia Journal of Agriculture and Forestry Science and Technology,56(7):31-34.(in Chinese)梁春波,黄绪堂,李岑,周菲,王文军,马军,陈惠荣,郭永利.2015.向日葵逆境应答转录因子DREB的克隆与表达分析.宁夏农林科技,56(7):31-34.
Liang Wei.2003.Investigation results of rust resistance of introduced chrysanthemum varieties in Lanzhou area and measures to prevent rust.Gansu Agricultural Science and Technology,(9):41-43.(in Chinese)梁伟.2003.兰州地区引进菊花品种的抗锈性调查结果及防锈病措施.甘肃农业科技,(9):41-43.
Licausi F,Ohme-Takagi M,Perata P.2013.APETALA2/ethylene responsive factor(AP2/ERF)transcription factors:mediators of stress responses and developmental programs.New Phytologist,199(3):639-649.
Liu L Q,Zhu K,Yang Y F,Wu J,Chen F D,Yu D Y.2008.Molecular cloning,expression profiling and trans-activationproperty studies of a DREB2-like gene from chrysanthemum(Dendranthema vestitum).Journal of Plant Research,121(2):215-226.
Liu Xiao-ying,Chen Li-yuan,Zhang Jing-qiu,Li Jia-wei,Gao Yue,Wang Zhen-ying.2013.Isolation and functional analysis of a new DREBtranscription factor(Bp DREB1)from Brassica pekinensis.Acta Agronomica Sinica,39(2):230-237.(in Chinese)刘晓颖,陈丽媛,张竞秋,李嘉玮,高越,王振英.2013.白菜脱水应答转录因子Bp DREB1基因的克隆及功能研究.作物学报,39(2):230-237.
Liu Zhi-wei,Xiong Yang-yang,Li Tong,Yan Ya-jun,Han Hong-run,Wu Zhi-jun,Zhuang Jing.2015.The cloning of transcription factor gene Cs DREB-A4 and the response to temperature stress in Camellia sinensis.Journal of Tea Science,35(1):24-34.(in Chinese)刘志薇,熊洋洋,李彤,严雅君,韩洪润,吴致君,庄静.2015.茶树转录子基因Cs DREB-A4的克隆与温度胁迫响应的分析.茶叶科学,35(1):24-34.
Lu J,Robert C A,Riemann M,Cosme M,Mène-SaffranéL,Massana J,Stout M J,Lou Y,Gershenzon J,Erb M.2015.Induced jasmonate signaling leads to contrasting effects on root damage and herbivore performance.Plant Physiology,167(3):1100-1116.
Nakano T,Suzuki K,Ohtsuki N,Tsujimoto Y,Fujimura T,Shinshi H.2006.Identification of genes of the plant-specific transcription-factor families cooperatively regulated by ethylene and jasmonate in Arabidopsis thaliana.Journal of Plant Research,119(4):407-413.
Pernas M,Sánchez-Monge R,Salcedo G.2000.Biotic and abioticstresses can induce cystatin expression in chestnut.Febs Letters,467(2-3):206-210.
Sakuma Y,Liu Q,Dubouzet J G,Abe H,Shinozaki K,Yamaguchi-Shinozaki K.2002.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression.Biochemical and Biophysical Research Communications,290:998-1009.
Sriram S,Chandran N K,Kumar R,Reddy M K.2015.First report of Puccinia horiana causing white rust of chrysanthemum in India.New Disease Reports,32:8.
Stockinger E J,Gilmour S J,Thomashow M F.1997.Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE,a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit.Proceedings of the National Academy of Sciences,94(3):1035-1040.
Tian Xiu-ling,Xia Bing-qiang,Luo feng-xia,Deng li-qin.1999.A preliminary study on the pathogen and infection pathway of white rust in Chrysanthemum.Journal of Shenyang Aricultural University,30(3):379-380.(in Chinese)田秀玲,夏炳强,罗凤霞,邓丽琴.1999.菊花白色锈病的病原菌和侵染途径的初步研究.沈阳农业大学学报,30(3):379-380.
Whipps J M.1993.A review of white rust(Puccinia horiana Henn.)disease on chrysanthemum and the potential for its biological control with Verticillium lecanii(Zimm.)Viégas.Annals of Applied Biology,122(1):173-187.
Xie Deng-lei,Cui Jiang-hui,Chang Jin-hua.2013.Cloning and expression analysis of Sb DREB2 gene from Sorghum bicolor.Acta Agronomica Sinica,39(8):1352-1359.(in Chinese)谢登雷,崔江慧,常金华.2013.高粱中Sb DREB2基因的克隆与表达分析作.作物学报,39(8):1352-1359.
Yang Ying-jie,Li Chun-shui,Zhang Xiao-jiao,Wei Qian,Gao Jun-ping,Hong Bo.2013.Drought stress tolerance of the hybrid offspring produced by At DREB1A transgenic chrysanthemum.Journal of Agricultural Biotechnology,21(2):148-157.(in Chinese)杨英杰,李春水,张晓娇,魏倩,高俊平,洪波.2013.转At DREB1A基因菊花杂交后代优株水分胁迫耐性分析.农业生物技术学报,21(2):148-157.
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