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渗调蛋白(osmotin,OSM)是植物中普遍存在的多功能蛋白,研究已明确OSM是胁迫响应蛋白,由参与胁迫响应的功能推测,OSM通过参与生长发育,调控植物对胁迫的响应,但OSM参与生长发育的研究报道极为有限。本研究在分离了马铃薯(Solanum tuberosum)StOSM-3b基因cDNA的基础上,用p Cambia1305-35S载体,通过基因工程技术,构建了抑制渗调蛋白表达的反义StOSM-3b基因cDNA载体,农杆菌(Agrobacterium tumefaciens)介导途径转化反义StOSM-3b至马铃薯体内,筛选获得β-葡萄糖苷酸酶(β-glucuronidase,GUS)染色阳性转化株系,阳性株系叶片StOSM-3b mRNA积累和渗调蛋白酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)定量数据分析显示,抑制株系叶片StOSM-3b mRNA积累是对照的25%,渗调蛋白含量是对照株系的52%,这表明反义StOSM-3b部分抑制了StOSM-3b表达,下调了马铃薯渗调蛋白的体内积累;下调渗调蛋白表达株系表现为生长缓慢、个体矮小、且生育期显著缩短、不能正常结薯的表型,且生长节奏失调。该研究结果为渗调蛋白参与生长和发育过程提供了直接证据。
Osmotin (OSM) is a ubiquitous multifunctional protein in plants. It has been well established that OSM is a stress response protein. It is speculated that the function of OSM is involved in the response to stress. OSM regulates plant response to stress by participating in growth and development, but OSM Research reports involved in growth and development are extremely limited. In this study, we isolated the cDNA of StOSM-3b from potato (Solanum tuberosum), and constructed the cDNA vector of antisense StOSM-3b gene that inhibits the expression of osmotic protein by using p Cambia1305-35S vector. (Agrobacterium tumefaciens) -mediated pathway to transform the antisense StOSM-3b into potato. The positive transformants stained with β-glucuronidase (GUS) were screened. The accumulation and infiltration of StOSM-3b mRNA in the leaves of positive plants Quantification of enzyme-linked immunosorbent assay (ELISA) showed that accumulation of StOSM-3b mRNA was 25% of the control and the amount of the osmotic protein was 52% of that of the control, indicating that antisense StOSM -3b partially inhibited the expression of StOSM-3b and down-regulated the in vivo accumulation of potato osmotic protein; down-regulated osmotic protein expression showed slow growth, short individuals and shortened growth period, Rhythm of growth disorders. The findings provide direct evidence that the osmotic proteins are involved in the growth and developmental processes.