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转病毒来源发夹RNA小麦表现对大麦黄矮病毒的抗性 被引量:6

Transgenic wheat expressing virus-derived hairpin RNA is resistant to Barley yellow dwarf virus
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摘要 将大麦黄矮病毒GPV株系的复制酶基因片段和CP基因片段构建成可在植物细胞内表达含有双链复制酶RNA(茎)和反义CPRNA(环)的复合发夹RNA结构,希望能够诱发植物体针对病毒的RNA干扰作用,从而达到抗病毒目的。利用基因枪法将该结构导入小麦幼胚愈伤组织细胞后,通过在幼苗再生阶段进行以叶片为模板的快速PCR来加速阳性植株的筛选过程,最终共获得基因组整合有外源基因的小麦再生植株21株。对再生植株接种不同剂量的病毒,其中9株对BYDV-GPV有低度抗性,表现在低接毒量时无症状,接毒量提高时发病且严重;6株具中度抗性,表现在低接毒量时无症状,接毒量提高时局部有不严重症状;6株具高度抗性,两种情况下均无症状。抗性实验结果表明,hpRNA介导对BYDV的抗性可能受到BYDV含量的影响,具有剂量效应的特点。 An expression vector expected to induce RNA interference against Barley yellow dwarf virus (BYDV), which expressed a composite hpRNA with the dsRNA stem homologous of BYDV GPV replicase gene and the antisense RNA loop homologous of coat protein gene, was designed without marker gene. The vector was transferred into callus cells from wheat (Triticum aestivum L.) immature embryos by particle bombardment. To select the positive transformants as early as possible, a rapid PCR, which does not need extract wheat DNA instead of few leaves, was used at regeneration stage of plantlets. Totally 21 plants proved to contain alien sequence. Antivirus test with high dose infected virus revealed that, 9 plants showed low level of resistance to BVDV, 6 plants showed moderate resistance and 6 plants showed high level of resistance. Interestingly, both low and moderate levels of resistance plants were no symptoms when infected by viruses at low dose. It suggests the dose- dependent effect of the resistance mediated by hpRNA to BYDV-GPV.
出处 《遗传》 CAS CSCD 北大核心 2007年第1期97-102,共6页 Hereditas(Beijing)
关键词 发夹RNA RNA干扰 大麦黄矮病毒 转基因 小麦 hairpin RNA RNA interference Barley yellow dwarf virus transgenic wheat
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参考文献14

  • 1燕飞,成卓敏.RNA干扰机制研究进展[J].遗传,2005,27(1):167-172. 被引量:7
  • 2Meister G,Tuschl T.Mechanisms of gene silencing by double-stranded RNA.Nature,2004,431(7006):343-349.
  • 3燕飞,成卓敏.病毒编码的转录后沉默抑制蛋白[J].生命的化学,2004,24(4):281-284. 被引量:4
  • 4Qu F, Morris TJ. Suppressors of RNA silencing encoded by plant viruses and their role in viral infections. FEBS Lett, 2005, 579(26): 5958--5964.
  • 5燕飞,成卓敏.RNA干扰技术在几项研究领域的应用[J].中国生物工程杂志,2004,24(8):14-19. 被引量:2
  • 6Wang MB, Abbott DC, Waterhouse PM. A single copy of a virus-derived transgene encoding hairpin RNA gives immunity to Barley yellow dwarf virus. Mol Plant Pathol,2000, 1(6): 347--356.
  • 7Tenllado F, Diaz-Ruiz JR. Double-stranded RNA-mediated interference with plant virus infection. J Virol, 2001,75(24): 12288-- 12297.
  • 8Kalantidis K, Psaradakis S, Tabler M, Tsagris M. The occurrence of CMV-specific short RNAs in transgenic tobacco expressing virus-derived double-stranded RNA is indicative of resistance to the virus. Mol Plant Microbe Interact, 2002, 15(8): 826--833.
  • 9Wang MB, Metzlaff M. RNA silencing and antiviral defense in plants. Curr Opin Plant Biol, 2005, 8(2): 216--222.
  • 10Cheng ZM, He XY, Chen CC, Zhang J, Xiao H, Zhou GH.Barley yellow dwarf virus coat protein gene and transgenic wheat plants obtained by pollen tube path way. Prog Nat Sci, 1994, 4(2): 235--240.

二级参考文献92

  • 1[4]Steiner J J,Poklemba C J,Fjellstyom R G, Elliott L F. A rapid one-tube genomic DNA extraction process for PCR and RAPD analyses[J]. Nucleic Acids Research,1995,23:2569~2570.
  • 2[6]Mocouch S R, Kohert G, Yu Z H, Wang Z Y, Khush G S,Coffi man W R, Tanksley S D. Molecular mapping of rice chromo- somes[J]. TAG, 1988,76 : 815 ~ 829.
  • 3[7]HE Guanghua,HOU Lei,XIAO Yuehua,LUO Xiaoying,NIU Guoqing, YANG Guangwei, PEI Yan. A common sequence dof ference between cytoplasmic male sterile lines and their main tainer lines existing in rice(Oryza satival. )chloroplast tRNA leu gene region[J]. Euphytica,2003,131(3):269~274.
  • 4[8]Sonneveld T,Robbins T P,Boskovic R,Tobutt K R. Cloning of six cherry self-incompatibility alleles and development of allele-specific PCR detection[J]. Theo Appl Gen,2001,102:1046~1058.
  • 5[9]Gu W K,Weeden N F,Yu J,Wallace D H. Large scale cost-ef fective screening of PCR products in marker-assisted selection applications[J]. Theor Appl Genet, 1995,91: 465~ 470.
  • 6Semizarov D, Frost L, Sarthy A, Kroeger P, Halbert D N, Fesik S W.Specificity of short interfering RNA determined through gene expression signatures.Proc Natl Acad Sci USA, 2003, 100(11):6347~6352.
  • 7Jackson A L, Bartz S R, Schelter J, Kobayashi S V, Burchard J, Mao M, Li B, Cavet G, Linsley P S.Expression profiling reveals off-target gene regulation by RNAi.Nat Biotechnol, 2003, 21(6):635~637.
  • 8Chi J T, Chang H Y, Wang N N, Chang D S, Dunphy N, Brown P O.Genomewide view of gene silencing by small interfering RNAs.Proc Natl Acad Sci USA, 2003, 100(11): 6343~6346.
  • 9Uhlirova M, Foy B D, Beaty B J, Olson K E, Riddiford L M, Jindra M.Use of Sindbis virus-mediated RNA interference to demonstrate a conserved role of Broad-Complex in insect metamorphosis.Proc Natl Acad Sci USA, 2003, 100(26):15607~15612.
  • 10Van der Giessen K, Di-Marco S, Clair E, Gallouzi I E.RNAi-mediated HuR depletion leads to the inhibition of muscle cell differentiation.J Biol Chem, 2003, 278(47):47119~47128.

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