CAJ | 학술논문

从烟草(Nicotiana tabacum L.)中克隆了3个病原物诱导性启动子PPP1、PPP2和PPP3,它们都含受细菌诱导的反应元件PPP1和PPP2中,另含有受生物激发子及水杨酸(SA)的诱导元件;PPP1内部还含重复的两段111bp的序列,而在PPP2K ,这个重复序列号中的一个111bpr的片段被定点删除.分别构建了含这三个启动子及花椰菜花叶病毒35S启动子的转化单元,用它们分别转化拟南芥(Arabidopsis thaliana L.)获得了转基因植株.PCR证明,几个启动子和所带的gus基因(uidA)已经整合到拟南芥基因组中.用青枯病菌接种转基因第二代植株,组织染色表明三个PPP启动子在拟南芥中都可以受青枯病菌诱导,说明克隆的PPP启动子是活性的.随后分别用SA、来自水稻白叶枯病菌的蛋白质激发子harpinx00以及harpinx00的3个具有不同功能的片段DEG(促进生长)、DIR(诱导抗病)喷雾处理转基因植株,通过GUS 的荧光定量分析,检测了启动子的活性.结果显示,表枯病菌诱导PPP1、PPP2和PPP3活性分别为35S启动子活性的53、39和25倍.PPP1和PPP2可以受SA、harpinx00、DEG、DIR和DPR的诱导,而PPP3则不能.这些结果说明了有关元件的可能作用.另外,PPP1 的活性比PPP2高3倍,表明11bp重复序列可以影响启动子活性水平.
종연초(Nicotiana tabacum L.)중극륭료3개병원물유도성계동자PPP1、PPP2화PPP3,타문도함수세균유도적반응원건PPP1화PPP2중,령함유수생물격발자급수양산(SA)적유도원건;PPP1내부환함중복적량단111bp적서렬,이재PPP2K ,저개중복서렬호중적일개111bpr적편단피정점산제.분별구건료함저삼개계동자급화야채화협병독35S계동자적전화단원,용타문분별전화의남개(Arabidopsis thaliana L.)획득료전기인식주.PCR증명,궤개계동자화소대적gus기인(uidA)이경정합도의남개기인조중.용청고병균접충전기인제이대식주,조직염색표명삼개PPP계동자재의남개중도가이수청고병균유도,설명극륭적PPP계동자시활성적.수후분별용SA、래자수도백협고병균적단백질격발자harpinx00이급harpinx00적3개구유불동공능적편단DEG(촉진생장)、DIR(유도항병)분무처리전기인식주,통과GUS 적형광정량분석,검측료계동자적활성.결과현시,표고병균유도PPP1、PPP2화PPP3활성분별위35S계동자활성적53、39화25배.PPP1화PPP2가이수SA、harpinx00、DEG、DIR화DPR적유도,이PPP3칙불능.저사결과설명료유관원건적가능작용.령외,PPP1 적활성비PPP2고3배,표명11bp중복서렬가이영향계동자활성수평.
Harpins are bacterial proteins that can enhance plant growth and detense against pathogens and insects. To elaborate whether harpins perform the diverse functions in coordination with the activation of specific promoters that contain particular elements, we cloned pathogen-inducible plant promoters PPP1, PPP2, and PPP3 from tobacco and investigated their responses to harpinXoo or its truncated fragments DEG, DIR, and DPR (domains for enhancing plant growth, insect resistance and pathogen resistance).PPP1 contains an internal repeat composed of two tandem 111 bp fragments; 111 bp in the repeat was deleted in PPP2. PPP3 contains a bacteria-inducible element; PPP1 and PPP2 additionally contain TAC-1 and Eli boxes inducible correspondingly by salicylic acid (SA) and elicitors. Function of cloned PPPs was confirmed based on their activation in transgenic Arabidopsis plants by Ralstonia solanacearum (Ralston)or SA. HarpinXoo, DEG, DIR, or DPR activated PPP1 and PPP2 but not PPP3, consistent with the presence of Eli boxes in promoters. PPP1 was ca. 3-fold more active than PPP2, suggesting that the internal repeat affects levels of the promoter activation.