接种不同AM真菌对滇重楼幼苗功能基因表达的影响

为探究接种不同外源性从枝菌根(Arbuscular mycorrhizae,AM)真菌对滇重楼幼苗基因表达的影响,本研究以灭菌土壤为生长基质,将滇重楼(Paris polyphylla var. yunnanensis)新鲜种子与28种AM真菌于室温盆栽条件下共培养,采用实时荧光定量PCR的方法检测鲨烯环氧酶基因(Squalene epoxidase,SE)、共生受体类似激酶基因(Symbiosis-receptor-like kinase,SYMRK)、产生钙离子振荡的通道蛋白基因(Doesn’t making fections 1,DMI1)、钙/钙调依赖性蛋白激酶基因(Calcium/calmodulin-dependent protein kinase,CCa MK)4个功能基因在滇重楼幼苗的差异表达情况。结果表明:28株AM菌株可不同程度的影响SE、SYMRK、DMI1、CCa MK 4个功能基因的表达,其中薄壁两性囊霉(Ambisporaleptoticha,Ale)和崔氏原囊霉(Archaeospora trappei,Atr)可以显著增加4个功能基因的表达量。隐类球囊霉(Paraglo-mus occultum,Po)和透明盾巨孢囊霉(Scutellospora pellucida,Spe)可以增加SE、SYMRK的表达,细凹无梗囊霉(Acau-lospora scrobiculata,Asc),亮色盾巨孢囊霉(Racocetra fulgida,Rfu),哥伦比亚内养囊霉(Entrophospora colombiana,Ec),明球囊霉(Rhizophagus clarus,Rcl),根内球囊霉(Rhizophagus intraradices,Rin)可以增加DMI1的表达。综上所述,接种的28株AM真菌菌株中Ale和Atr能显著的提高4种功能基因的表达,结合前期与滇重楼无菌播种幼苗进行共生培养分析AM真菌对种子萌发及幼苗化学成分的影响,推测这两株菌可望能作为培育滇重楼菌根化苗的理想菌株,人工接种AM真菌可为保护和提高滇重楼开辟了一条新的途径。

Effects of inoculation with different AM fungi on functional gene expression in seedlings of Paris polyphylla var. yunnanensis

The aim of this study was to investigate the expression of function gene in the seedling of Paris polyphylla var.yunnanensis inoculated by different foreign arbuscular mycorrhizal(AM)fungi species.With sterilized soil as the growth of the substrate,the fresh seeds of P.polyphylla var.yunnanensis were co-cultured with 28 AM fungi species under the condition of pot culture at room temperature.The differential expression of four functional genes including Squalene epoxidase(SE),Symbiosisreceptor-like kinase(SYMRK),Doesn’t making fections 1(DMI1),Calcium/calmodulin-dependent protein kinase(CCaMK),were detected by real-time fluorescence quantitative PCR.The results showed that the 28 strains of AM strains could affect the expression of four functional genes SE,SYMRK,DMI1 and CCaMK in different extent.Ambispora leptoticha(Ale)and Archaeospora trappei(Atr)can significantly increase the expression of the four functional genes.Paraglomus occultum,(Po)and Scutellospora pellucida(Spe)can increase the expression of SE,SYMRK;Acaulospora scrobiculata(Asc),Racocetra fulgida(Rfu),Entrophospora colombiana(Ec),Rhizophagus clarus(Rcl)and Rhizophagus intraradices(Rin)can increase the expression of DMI1.In conclusion,the expression of four functional genes was significantly increased by Ale and Atr in 28 strains of AM fungi.There was obvious influence that the seed germination and seedling chemical constituents of Paris polyphylla var.yunnanensis inoculated by different foreign arbuscular mycorrhizal fungi species in our previous study.It is presumed that these two strains(Ale and Atr)are expected to serve as the ideal strains for the mycorrhizal seedlings cultivation of Paris polyphylla var.yunnanensis.Artificial inoculation of AM fungi can open up a new way for the protection and improvement of Paris polyphylla var.yunnanensis.