Study On Theexpressionofgene Associatedwith Sympathy In Wheat Indocing Para-Nodules

Leguminous plants and rhizobial bacteria symbiotically generate nodules,thereby carrying out symbiotic nitrogen fixation,which is the most important form of biological nitrogen fixation.Rhizobia produce nodulation factor(NF)after being stimulated by plant flavonoids,NF binds to specific receptors on the root surface of legumes,auses inter-conduction of intracellular calcium signal and other series of signals.Eventually lead to the formation of infestation lines and eventually nodule formation.The intracellular conduction pathway that the root cells of plants respond to rhizobia nodulation factors and eventually produce root nodules is the SYM pathway.The symbiotic signal pathway exists in wheat and rice and other gramineous plants,and can form symbiotic interactions with arbuscular mycorrhizal fungi through some signal molecules in the pathway.The present study compared the effects of 2,4-D,flavonoids,chloroanthraquinone,and Doxorubicin on induction of nodulation in wheat and analyzed the changes inwheat physiological parameters after adding inducer.At the same time,we are using bioinformatics data comparison and real-time fluorescence quantitative analysis,we analyzed and verified the nodulation signal recognition genes TaNSP1,Nodulation factor receptor kinase TaNFR1 and TaNFR5 and auxin transport factors TaPIN1 in the wheat SYM pathway with addition of 2,4-D and flavonoids.Chlorsulform and Chloramben four kinds of inducers when their expression levels change.The experimental results show that all four inducers can induce wheat nodule production.Lateral comparisons showed that the efficiency of wheat nodules induced by2,4-D and chloroanthraquinone was higher,but Chlorsulform had a greater effect on wheat growth and development.The other two inducers induce low nodulation efficiency.In comprehensive comparison,2,4-D is more suitable as an inducer used in actual production.The other two inducers induce low nodulation efficiency.In comprehensive comparison,2,4-D is more suitable as an inducer used in actual production.Using laser confocal scanning microscopy to observe the rhizobial colonization of root nodules produced by wheat after inducer and found that Azorhizobium caulinodans ORS571 can colonize wheat.The RT-qPCR technique was used to detect the relative expression levels of the upstream nodulation recognition genes TaNSP1,Nodulation factor receptor kinase TaNFR5,TaNFR1 and downstream auxin transporter TaPIN1 in wheat SYM pathway.The results showed that after the inducer was added without addition of Rhizobium,the relative expression levels of the auxin transporter TaPIN1 were increased to a certain extent except that induced by 2,4-D.After the addition of rhizobia,the expression of each gene was up-regulated in different ranges.The above results indicate that the addition of inducer does not activate the relevant genes in the SYM pathway,and that only auxin stimulated the expression of auxin transport-related genes.After the addition of rhizobia,relative expression levels of key genes TaPIN1 and nodulation receptor receptor kinases TaNFR1,TaNFR5 and the nodulation signal recognition genes TaNSP1 were up-regulated on the SYM pathway,indicating that inducers and Azotobacter inducing the expression of related genes on the wheat SYM pathway.It shows that the related genes in wheat SYM pathway are affected by the inducer and rhizobia.