目的克隆猪苓Polyporus umbellatus菌核无机磷酸盐转运蛋白基因并进行生物信息学和表达模式分析。方法采用RT-PCR技术从猪苓菌核中克隆得到1个无机磷酸盐转运蛋白(inorganic phosphate transporter)基因,利用生物信息学软件预测蛋白的...目的克隆猪苓Polyporus umbellatus菌核无机磷酸盐转运蛋白基因并进行生物信息学和表达模式分析。方法采用RT-PCR技术从猪苓菌核中克隆得到1个无机磷酸盐转运蛋白(inorganic phosphate transporter)基因,利用生物信息学软件预测蛋白的理化性质、结构域、信号肽和跨膜结构等分子特性;采用Clustal W2以及MEGA 6.0分别进行氨基酸多序列比对和进化关系分析;实时荧光定量PCR分析基因表达模式。结果猪苓无机磷酸盐转运蛋白基因命名为Pu Pi T(NCBI登录号:KU179154)。该基因开放读码框全长为1 590 bp,编码530个氨基酸。Pu Pi T蛋白相对分子质量为57 552,等电点6.82。氨基酸序列分析表明,Pu Pi T蛋白具有12个跨膜区。氨基酸序列多重比对及系统发育树结果显示Pu Pi T与Moniliophthora rorer亲缘关系最近,与Moniliophthora roreri、双色蜡蘑Laccaria bicolor、Heterobasidion irregulare有较高的同源性,荧光定量PCR结果表明,Pu Pi T在与蜜环菌共生的猪苓菌核及未与蜜环菌共生的菌核中都有表达,其中共生部分的表达量显著上调,约是未共生部位的12倍,说明其参与猪苓与蜜环菌共生过程。结论 Pu Pi T基因克隆和分子特征为进一步研究揭示其在猪苓菌核磷元素转运及与蜜环菌共生过程中的调控作用奠定基础。展开更多
Arbscular mycorrhiza(AM),a symbiosis between plants and members of fungi Glomeromycota,improves the resistance,nutrition and material metabolism of plant.Arbuscules generated by symbiotic development,is the main site ...Arbscular mycorrhiza(AM),a symbiosis between plants and members of fungi Glomeromycota,improves the resistance,nutrition and material metabolism of plant.Arbuscules generated by symbiotic development,is the main site of nutrient exchange and genetic material recombination,resulting in physiological changes and gene expression regulation.AM regulates pathogenesis-related protein(PRs) genes and antioxidant enzyme genes against biotic and abiotic stresses.Nutrient exchange induced by AM is directly involved in uptake,transformation and utilization of nutrient elements in plants.Importantly,transporter genes play an important role in phosphate,nitrogen and carbon acquisition.In AM interactions,phosphate transporter(PT) genes,from both arbuscular mycorrhizal fungi and plant root,are induced and there product promote phosphate acquisition;increasing expression of ammonium transporter(AMT) genes and arginine biosynthesis/degradation enzyme genes is in charge of nitrogen acquisition;and promotion mechanism of carbon acquisition is involved in up-regulation of sugar transporter genes.In addition,secondary metabolites,functioned as signal moleculars and defense compounds,are increased with development of AM symbiont by up-regulating related synthetic genes based on different promotion mechanism.Taken together,molecular regulation of plant and arbuscular mycorrhizal fungi,induced by infection process,stimulate plant nutrient acquisition and resist to biotic/abiotic stress.展开更多
文摘目的克隆猪苓Polyporus umbellatus菌核无机磷酸盐转运蛋白基因并进行生物信息学和表达模式分析。方法采用RT-PCR技术从猪苓菌核中克隆得到1个无机磷酸盐转运蛋白(inorganic phosphate transporter)基因,利用生物信息学软件预测蛋白的理化性质、结构域、信号肽和跨膜结构等分子特性;采用Clustal W2以及MEGA 6.0分别进行氨基酸多序列比对和进化关系分析;实时荧光定量PCR分析基因表达模式。结果猪苓无机磷酸盐转运蛋白基因命名为Pu Pi T(NCBI登录号:KU179154)。该基因开放读码框全长为1 590 bp,编码530个氨基酸。Pu Pi T蛋白相对分子质量为57 552,等电点6.82。氨基酸序列分析表明,Pu Pi T蛋白具有12个跨膜区。氨基酸序列多重比对及系统发育树结果显示Pu Pi T与Moniliophthora rorer亲缘关系最近,与Moniliophthora roreri、双色蜡蘑Laccaria bicolor、Heterobasidion irregulare有较高的同源性,荧光定量PCR结果表明,Pu Pi T在与蜜环菌共生的猪苓菌核及未与蜜环菌共生的菌核中都有表达,其中共生部分的表达量显著上调,约是未共生部位的12倍,说明其参与猪苓与蜜环菌共生过程。结论 Pu Pi T基因克隆和分子特征为进一步研究揭示其在猪苓菌核磷元素转运及与蜜环菌共生过程中的调控作用奠定基础。
基金financed by the Doctoral Foundation of Ministry of Education of China(20110101110094)
文摘Arbscular mycorrhiza(AM),a symbiosis between plants and members of fungi Glomeromycota,improves the resistance,nutrition and material metabolism of plant.Arbuscules generated by symbiotic development,is the main site of nutrient exchange and genetic material recombination,resulting in physiological changes and gene expression regulation.AM regulates pathogenesis-related protein(PRs) genes and antioxidant enzyme genes against biotic and abiotic stresses.Nutrient exchange induced by AM is directly involved in uptake,transformation and utilization of nutrient elements in plants.Importantly,transporter genes play an important role in phosphate,nitrogen and carbon acquisition.In AM interactions,phosphate transporter(PT) genes,from both arbuscular mycorrhizal fungi and plant root,are induced and there product promote phosphate acquisition;increasing expression of ammonium transporter(AMT) genes and arginine biosynthesis/degradation enzyme genes is in charge of nitrogen acquisition;and promotion mechanism of carbon acquisition is involved in up-regulation of sugar transporter genes.In addition,secondary metabolites,functioned as signal moleculars and defense compounds,are increased with development of AM symbiont by up-regulating related synthetic genes based on different promotion mechanism.Taken together,molecular regulation of plant and arbuscular mycorrhizal fungi,induced by infection process,stimulate plant nutrient acquisition and resist to biotic/abiotic stress.
