Wang Keri, Conn Kenneth, Lazarovits George
Southern Crop Protection and Food Research Center, Agriculture and Agri-Food Canada, 1391 Sandford St., London, ON, Canada N5V 4T3.
Appl Environ Microbiol. 2006 Jan;72(1):760-8. doi: 10.1128/AEM.72.1.760-768.2006.
Burkholderia sp. strain PsJN stimulates root growth of potato explants compared to uninoculated controls under gnotobiotic conditions. In order to determine the mechanism by which this growth stimulation occurs, we used Tn5 mutagenesis to produce a mutant, H41, which exhibited no growth-promoting activity but was able to colonize potato plants as well as the wild-type strain. The gene associated with the loss of growth promotion in H41 was shown to exhibit 65% identity at the amino acid level to the nadC gene encoding quinolinate phosphoribosyltransferase (QAPRTase) in Ralstonia solanacearum. Complementation of H41 with QAPRTase restored growth promotion of potato explants by this mutant. Expression of the gene identified in Escherichia coli yielded a protein with QAPRTase activities that catalyzed the de novo formation of nicotinic acid mononucleotide (NaMN). Two other genes involved in the same enzymatic pathway, nadA and nadB, were physically linked to nadC. The nadA gene was cotranscribed with nadC as an operon in wild-type strain PsJN, while the nadB gene was located downstream of the nadA-nadC operon. Growth promotion by H41 was fully restored by addition of NaMN to the tissue culture medium. These data suggested that QAPRTase may play a role in the signal pathway for promotion of plant growth by PsJN.
与无菌条件下未接种的对照相比,伯克霍尔德氏菌属菌株PsJN能刺激马铃薯外植体的根系生长。为了确定这种生长刺激发生的机制,我们使用Tn5诱变产生了一个突变体H41,该突变体没有生长促进活性,但能够像野生型菌株一样定殖于马铃薯植株。结果表明,与H41中生长促进作用丧失相关的基因在氨基酸水平上与茄科劳尔氏菌中编码喹啉酸磷酸核糖基转移酶(QAPRTase)的nadC基因具有65%的同一性。用QAPRTase对H41进行互补恢复了该突变体对马铃薯外植体的生长促进作用。在大肠杆菌中鉴定出的该基因的表达产生了一种具有QAPRTase活性的蛋白质,该蛋白质催化了烟酸单核苷酸(NaMN)的从头合成。参与同一酶促途径的另外两个基因nadA和nadB与nadC在物理上相连。在野生型菌株PsJN中,nadA基因与nadC作为一个操纵子共转录,而nadB基因位于nadA-nadC操纵子的下游。通过向组织培养基中添加NaMN,H41的生长促进作用完全恢复。这些数据表明,QAPRTase可能在PsJN促进植物生长的信号通路中发挥作用。