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草业学报  2014 

马铃薯SGT3基因表达及其启动子功能分析

DOI: 10.11686/cyxb20140224, PP. 196-206

Keywords: SGT3,基因表达,启动子,功能分析

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Abstract:

鼠李糖转移酶(rhamnosyltransferaseSGT3)是植物糖苷生物碱合成的关键酶,它主要调控α-茄碱和α-查茄碱从其β形式的转化。本文研究马铃薯栽培种SGT3基因的表达特点及其启动子的功能。实时定量PCR结果发现在红光照射24h后,SGT3的表达量是黑暗处理的26.8倍,说明红光显著诱导了SGT3的表达;为进一步分析该基因的光调控机理,本项研究克隆到SGT3上游长度为2449bp的启动子序列,通过分析发现该启动子的转录起始位点位于翻译起始位点上游-152bp,同时确定了该启动子核心序列及上游增强子、抑制子序列,受病原菌、损伤、干旱、ABA激素及一系列光调控的顺式元件。构建不同长度(349,572,979,1312和1870bp)的该启动子驱动报告基因GUS的植物表达载体并转化烟草。结果证明,不同长度的SGT3启动子都可以启动GUS表达,但没有CMV35S启动的GUS表达量高;其中在P572和P979的表达强度较高,这可能与该片段含有启动子的正调控元件(GATABOX,5′UTRPY-RICHSTRETCH)有关,GUS表达强度在P1312和P1870中明显减弱,预测到该区段存在抑制基因表达的负调控元件(WRKY710S);SGT3启动子的组织特异性实验表明GUS染色主要集中在烟草叶片的叶脉部分,茎中的表皮、韧皮部及木质部,但髓部几乎不表达,根中主要分布在根冠、分生区以及维管束组织中。上述结果为将来研究SGT3在糖苷生物碱合成过程中的调节功能提供了依据。

References

[1]  Xie Z, Allen E, Wilken A. DICER-LIKE 4 functions in trans-acting small interfering RNA biogenesis and vegetative phase change in Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the USA, 2005, 102(36): 12984-12989.
[2]  Detling J K. Mammalian herbivores: ecosystem-level effects in two grassland national parks. Wildlife Society Bulletin, 1998, 26: 438-448.
[3]  McCue K, Allen P, Shephers L. The primary in vivo steroidal alkaloid glucosyltransferase from potato. Phytochemistry, 2006, 67: 1590-1597.
[4]  Daraselia N D, Tarchevskaya S, Narita J O. The promoter for tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase gene 2 has unusual regulatory elements that direct high-level expression. Plant Physiology, 1996, 112: 727-733.
[5]  Rockhold D R, Chang S, Taylor N, et al. Structure of two Solanum bulbocastanum polyubiquitin genes and expression of their promoters in transgenic potatoes. American Journal of Potato Research, 2008, 85: 219-226.
[6]  安惠惠, 马晖玲, 李坚, 等. 农杆菌介导的Lyz-GFP基因对匍匐翦股颖Penn A-1转化和表达的研究. 草业学报, 2012, 21(3): 141-148.
[7]  van Staalduinen M A, Werger M J A. Marmot disturbances in a Mongolian steppe vegetation. Journal of Arid Environments, 2007, 69: 344-351.
[8]  David G G, Tony E G, Janice N, et al. Measurement of DNA copy number at microsatellite loci using quantitative PCR analysis. Cancer Research, 2000, 60: 5405-5409.
[9]  Nakashima K, Fujita Y, Katsura K, et al. Transcriptional regulation of ABI3-and ABA-responsive genes including RD29B and RD29A in seeds, germinating embryos, and seedlings of Arabidopsis. Plant Molecular Biology, 2006, 60: 51-68.
[10]  张宁, 王蒂. 农杆菌介导的烟草高效遗传转化体系研究. 甘肃农业科技, 2004, 9: 11-13.
[11]  张宁. 应用甜菜碱醛脱氢酶基因工程提高马铃薯抗逆性的研究. 兰州: 甘肃农业大学, 2004.
[12]  Eviner V T, Chapin III F S. Selective gopher disturbance influences plant species effects on nitrogen cycling. Oikos, 2005, 109: 154-166.
[13]  段光明, 刘加, 李霞. 马铃薯糖苷生物碱的生物学作用及开发利用. 资源开发与市场, 1995, 11(2): 61-65.
[14]  Elmayan T, Tepfer M. Evaluation in tobacco of theorgan specificity and strength of the rolD promoter, domain A of the 35S promoter and the 35S2 promoter. Transgenic Research, 1995, 4: 388-396.
[15]  Keddie J S, Hubner G, Slocombe S P, et al. Cloning and characterisation of an oleosin gene from Brassica napus. Plant Molecular Biology, 1992, 19: 443-453.
[16]  Jones C C, Halpern C B, Niederer J. Plant succession on gopher mounds in western cascade meadows: consequences for species diversity and heterogeneity. American Midland Naturalist, 2008, 159: 275-286.
[17]  Shirsat A, Wilford N, Croy R, et al. Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco. Molecular & General Genetics, 1989, 215: 326-331.
[18]  Xu X, Chen C, Fan B, et al. Physical and functional interactions between pathogen-induced Arabidopsis WRKY18, WRKY40, and WRKY60 transcription factors. Plant Cell, 2006, 18: 1310-1326.
[19]  Reichman O J, Seabloom E W. The role of pocket gophers as subterranean ecosystem engineers. Trends in Ecology and Evolution, 2002, 17: 44-49.
[20]  Reyes J C, Muro-Pastor M I, Florencio F J. The GATA family of transcription factors in Arabidopsis and rice. Plant Physiology, 2004, 134(4): 1718-1732.
[21]  Luo H, Song F, Goodman R M, et al. Up-regulation of OsBIHDI, a rice gene encoding BELL homeodomain transcriptional factor, in disease resistance responses. Plant Biology, 2005, 7(5): 459-468.
[22]  Canals R M, Herman D J, Firestone M K. How disturbance by fossorial mammals alters N cycling in a California annual grassland. Ecology, 2003, 84: 875-881.
[23]  Xie Z, Allen E, Wilken A. DICER-LIKE 4 functions in trans-acting small interfering RNA biogenesis and vegetative phase change in Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the USA, 2005, 102(36): 12984-12989.
[24]  Diaz-De-Leon F, Klotz K L, Lagrimini M. Nucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene. Plant Physiology, 1993, 101: 1117-1118.
[25]  Wright J P, Jones C G. Predicting effects of ecosystem engineers on patch-scale species richness from primary productivity. Ecology, 2004, 85: 2071-2081.
[26]  张堰铭, 刘季科. 地下鼠生物学特征及其在生态系统中的作用. 动物学报, 2002, 22: 144-154.
[27]  Daraselia N D, Tarchevskaya S, Narita J O. The promoter for tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase gene 2 has unusual regulatory elements that direct high-level expression. Plant Physiology, 1996, 112: 727-733.
[28]  Abe H, Tatsuno I, Tobe T, et al. Bicarbonate ion stimulates the expression of locus of enterocyte effacement-encoded genes in enterohemorrhagic Escherichia coli O157:H7. Infection and Immunity, 2002, 70(7): 3500-3509.
[29]  Zhang Y, Zhang Z, Liu J. Burrowing rodents as ecosystem engineers: the ecology and management of plateau zokors Myospalax fontanierii in alpine meadow ecosystems on the Tibetan plateau. Mammal Review, 2003, 33: 284-294.
[30]  Abe H, Urao T, Ito T, et al. Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell, 2003, 15: 63-78.
[31]  Nakashima K, Fujita Y, Katsura K, et al. Transcriptional regulation of ABI3-and ABA-responsive genes including RD29B and RD29A in seeds, germinating embryos, and seedlings of Arabidopsis. Plant Molecular Biology, 2006, 60: 51-68.
[32]  Fan N, Zhou W, Wei W, et al. Rodent pest management in the Qinghai-Tibet alpine meadow ecosystem. In: Singleton G R, Hinds L A, Leirs H, et al (eds.). Ecologically-based Management of Rodent Pests. ACIAR Monograph No. 59. Canberra: Australian Centre for International Agricultural Research, 1999: 285-304.
[33]  王权业, 周文扬, 张堰铭, 等. 高原鼢鼠挖掘行为观察. 动物学报, 1994, 14: 203-208.
[34]  Urao T, Yamaguchi-Shinozaki K, Urao S, et al. An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell, 1993, 5: 1529-1539.
[35]  Elmayan T, Tepfer M. Evaluation in tobacco of theorgan specificity and strength of the rolD promoter, domain A of the 35S promoter and the 35S2 promoter. Transgenic Research, 1995, 4: 388-396.
[36]  Gubler F, Jacobsen J V. Gibberellin-responsive elements in the promoter of a barley high-pI alpha-amylase gene. Plant Cell, 1992, 4(11): 1435-1441.
[37]  Díaz S, Cabido M. Vive la différence: plant functional diversity matters to ecosystem processes. Trends in Ecology and Evolution, 2001, 16: 646-655.
[38]  Xu X, Chen C, Fan B, et al. Physical and functional interactions between pathogen-induced Arabidopsis WRKY18, WRKY40, and WRKY60 transcription factors. Plant Cell, 2006, 18: 1310-1326.
[39]  Terzaghi W B, Cashmore A R. Light-regulated transcription. Annual Review of Plant Physiology and Plant Molecular Biology, 1995, 46: 445-474.
[40]  Abe H, Tatsuno I, Tobe T, et al. Bicarbonate ion stimulates the expression of locus of enterocyte effacement-encoded genes in enterohemorrhagic Escherichia coli O157:H7. Infection and Immunity, 2002, 70(7): 3500-3509.
[41]  Bonet A. Secondary succession of semi-arid Mediterranean old-fields in south-eastern Spain: insights for conservation and restoration of degraded lands. Journal of Arid Environments, 2004, 56: 213-233.
[42]  Thiesen H J, Bach C. Target Detection Assay (TDA): a versatile procedure to determine DNA binding sites as demonstrated on SP1 protein. Nucleic Acids Research, 1990, 18(11): 3203-3209.
[43]  Abe H, Urao T, Ito T, et al. Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell, 2003, 15: 63-78.
[44]  Jiang X L, Zhang W G, Wang G. Effects of different components of diversity on productivity in artificial plant communities. Ecological Research, 2007, 22: 629-634.
[45]  何俊龄. 土壤种子库与鼢鼠土丘植被恢复的关系. 兰州: 兰州大学, 2007.
[46]  Symstad A J. A test of the effects of functional group richness and composition on grassland invisibility. Ecology, 2000, 81: 99-109.
[47]  Urao T, Yamaguchi-Shinozaki K, Urao S, et al. An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell, 1993, 5: 1529-1539.
[48]  Logemann E, Parniske M, Hahlbrock K. Modes of expression and common structural features of the complete phenylalanine ammonia-lyase gene family in parsley. Proceedings of the National Academy of Sciences of the USA, 1995, 92(13): 5905-5909.
[49]  Sherrod A K, Seastedt T R. Effects of the northern pocket gopher (Thomomys talpoides) on alpine soil characteristics. Biogeochemistry, 2001, 55: 195-218.
[50]  Gubler F, Jacobsen J V. Gibberellin-responsive elements in the promoter of a barley high-pI alpha-amylase gene. Plant Cell, 1992, 4(11): 1435-1441.
[51]  Thum K E, Kim M, Christopher D A, et al. Cryptochrome 1, cryptochrome 2, and phytochrome a co-activate the chloroplast psbD blue light-responsive promoter. Plant Cell, 2001, 13(12): 2747-2760.
[52]  Odum E P. The strategy of ecosystem development. Science, 1969, 164: 262-270.
[53]  Morita A, Umemura T, Kuroyanagi M, et al. Functional dissection of a sugar-repressed α-amylase gene (Ramy1A) promoter in rice embryos. Febs Lett, 1998, 423: 81-85.
[54]  Chan C S, Guo L, Shih M C. Promoter analysis of the nuclear gene encoding the chloroplast glyceraldehyde-3-phosphate dehydrogenase B subunit of Arabidopsis thaliana. Plant Molecular Biology, 2001, 46 (2), 131-141.
[55]  邢智峰, 张安世, 徐存栓, 等. 毛白杨4CL基因启动子的克隆及初步功能分析. 河南师范大学学报, 2007, 35: 142-145.
[56]  季彦林, 王旺田, 王蒂. 不同光质对马铃薯块茎糖苷生物碱积累的诱导效应. 江苏农业学报, 2010, 26(1): 40-45.
[57]  Shirsat A, Wilford N, Croy R, et al. Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco. Molecular & General Genetics, 1989, 215: 326-331.
[58]  牛继平, 张金文, 王旺田, 等. 马铃SGAs合成代谢途径末端SGT酶基因克隆及序列分析. 草业学报, 2012, 21(3): 106-116. 浏览
[59]  Fields M J, Coffin D P, Gosz J R. Burrowing activities of kangaroo rats and patterns in plant species dominance at a shortgrass steppe-desert grassland ecotone. Journal of Vegetation Science, 1999, 10: 123-130.
[60]  Reyes J C, Muro-Pastor M I, Florencio F J. The GATA family of transcription factors in Arabidopsis and rice. Plant Physiology, 2004, 134(4): 1718-1732.
[61]  McCue K, Allen P V, Shepherd L V T, et al. Potato glycosterol rhamnosyltransferase, the terminal step in triose side-chain biosynthesis. Phytochemistry, 2007, 68: 327-334.
[62]  Archer S, Garrett M G, Detling J K. Rates of vegetation change associated with prairie dog (Cynomys ludovicianus) grazing in North American mixed-grass prairie. Vegetatio, 1987, 72: 159-166.
[63]  Morita A, Umemura T, Kuroyanagi M, et al. Functional dissection of a sugar-repressed α-amylase gene (Ramy1A) promoter in rice embryos. Febs Lett, 1998, 423: 81-85.
[64]  Hooper D U, Ewel J J, Hector A, et al. Effects of biodiversity on ecosystem functioning: a consensus of current knowledge and needs for future research. Ecological Monograph, 2005, 75: 3-35.
[65]  Luo H, Song F, Goodman R M, et al. Up-regulation of OsBIHDI, a rice gene encoding BELL homeodomain transcriptional factor, in disease resistance responses. Plant Biology, 2005, 7(5): 459-468.
[66]  Tatematsu K, Ward S, Leyser O, et al. Identification of cis-elements that regulate gene expression during initiation of axillary bud outgrowth in Arabidopsis. Plant Physiology, 2005, 138(2): 757-766.
[67]  Ward S E, Bardgett R D, McNamara N P, et al. Plant functional group identity influences short-term peatland ecosystem carbon flux: evidence from a plant removal experiment. Functional Ecology, 2009, 23: 454-462.
[68]  Brown C. Are functional guilds more realistic management units than individual species for restoration Weed Technology, 2004, (Supplement): 262-267.
[69]  Diaz-De-Leon F, Klotz K L, Lagrimini M. Nucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene. Plant Physiology, 1993, 101: 1117-1118.
[70]  陈婷婷, 杨青川, 丁旺, 等. 紫花苜蓿WRKY转录因子基因的克隆与亚细胞定位. 草业学报, 2012, 21(4): 159-167. 浏览
[71]  Connell J H. Diversity in tropical rain forests and coral reefs-high diversity of trees and corals is maintained only in a non-equilibrium state. Science, 1978, 199: 1302-1310.
[72]  Tatematsu K, Ward S, Leyser O, et al. Identification of cis-elements that regulate gene expression during initiation of axillary bud outgrowth in Arabidopsis. Plant Physiology, 2005, 138(2): 757-766.
[73]  Millar A, Kay S. Integration of circadian and photo transduction pathways in the network controlling CAB gene transcription in Arabidopsis. Proceedings of the National Academy of Sciences of the USA, 1996, 93: 15491-15496.
[74]  Rogers W E, Hartentt D C, Elder B. Effects of plains pocket gopher (Geomys bursarius) disturbances on tallgrass-prairie plant community structure. The American Midland Naturalist Journal, 2001, 145: 344-357.
[75]  Terzaghi W B, Cashmore A R. Light-regulated transcription. Annual Review of Plant Physiology and Plant Molecular Biology, 1995, 46: 445-474.
[76]  Terzaghi W, Cashmore A. Light-regulated transcription. Annual Review of Plant Physiology and Plant Molecular Biology, 1995, 46: 445-474.
[77]  Sherrod S K, Seastedt T R, Walker M D. Northern pocket gopher (Thomomys talpoides) control of alpine plant community structure. Arctic, Antarctic, and Alpine Research, 2005, 37: 585-590.
[78]  Thiesen H J, Bach C. Target Detection Assay (TDA): a versatile procedure to determine DNA binding sites as demonstrated on SP1 protein. Nucleic Acids Research, 1990, 18(11): 3203-3209.
[79]  Degenhardt J, Tobin E. A DNA binding activity for one of two closely defined phytochrome regulatory elements in an Lhcb promoter is more abundant in etiolated than in green plants. Plant Cell, 1996, 8: 31-41.

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