The Correlation Gene Clone And SNP Polymorphism Analysis Of The Stomatal Traits In Rice

Stomata is distributed on the surface of the air-borne organs of plants,and is an important channel of gas exchange between plants and the environment.Carbonic anhydrase(CA1,CA4),FLP and some genes which act on the EPF-TMM-SDD approach are considered to be important regulator affecting the stomatal character of plants.Japonica rice and indica rice are two subspecies of Rice,and they have great significance in stomatal character difference.However,the mechanism leading to the difference is still unclear.We isolate genes related to stomatal traits in rice,It is helpful to further understand the molecular mechanism of stomatal character development in rice.In this study,japonica rice and indica rice were used as research materials.Then six genes about stoma OsCA1,OsCA4,OsFLP OsSTO,OsTMM,and OsSDD1 were cloned by the method of homologous cloning and comparative genomics.We analyze the amino acid structure and molecular evolution.In addition,we identify SNP sites in japonica and indica rice,and verify some genes function in overexpression.The main results as follows:The OsCA1 gene with 819 bp is from chromosome 1,which encodes 273 amino acids.The genes from japonica rice and indica rice are identical in length.The length of the OsCA4 gene is 1002 bp,which encodes 344 amino acids.The results of gene comparison between japonica rice and indica rice showed that,the 273 th base of Os CA4 gene in japonica rice is T,but it is C in indica rice,The 504 th base of this gene in japonica rice is G,while it is C in indica rice.The mutation from G to C does not resulted in the change of relative amino acids.The total length of OsFLP gene is 1617 bp,which encodes 539 amino acids.The genes from japonica rice and indica rice are identical in length.The 1396 th base of Os FLP gene in japonica rice is C,but it is A in indica rice.The mutation from G to C resulted in the change of relative amino acid,The 466 th amino acid is R(arginine)in japonica rice,while it is S in indica rice.The OsSTO gene with 896 bp is from chromosome 1,and coding region lenth is 372 bp,it encodes 124 amino acids.The results of gene comparison between japonica rice and indica rice showed that,the 283 th base of Os STO gene in japonica rice is T,but it is C in indica rice,the 300 th base of this gene in japonica rice is A,while it is T in indica rice,but it’s all found on introns.The OsTMM gene with 1485 bp is from chromosome 1,which encodes 494 amino acids.The results of gene comparison between japonica rice and indica rice showed that,the 173 th base of OsTMM gene in japonica rice is T,but it is C in indica rice;The 1194 th base of this gene in japonica rice is C,while it is G in indica rice.The mutation from C to G resulted in the change of relative amino acid,it is Q(glutamine)in japonica rice,while it is E(glutamate)in indica rice.The OsSDD1 gene with 2192 bp is from chromosome 12,which encodes 733 amino acids.The results of gene comparison between japonica rice and indica rice showed that,the 414 th base of Os SDD1 gene in japonica rice is A,but it is G in indica rice.The mutation from A to G resulted in the change of relative amino acid,it is S(serinein)in japonica rice,while it is G(glycine)in indica rice.In this study,we also construct eukaryotic expression vector for genes related to the EPF-TMM-SDD1 pathway.To get the corresponding transgenic plants,we construct the eukaryotic expression vector of OsSTO,OsTMM,OsSDD1 gene,and import to Nipponbare using the method of Agrobacterium-mediated transformation.We determine the stomatal characters of transgenic plants and found that: Overexpression of the related genes in EPF-TMM-SDD approach,they decrease the stomatal density of transgenic plants,but they increase the stomatal length,while the stomata width is not significant,and only SDD1 gene decreases stomatal width slightly,it is proved that the cloned gene has the function of stomatal characters,and it lays a theoretical foundation for further analyzing the molecular mechanism of stomatal difference between japonica rice and indica rice.