The Function Of Orphan Gene OsCPY In Rice Growth Promotion

In the early stage of the laboratory,a new gene was found in a cDNA library of drought and salt-stressed rice roots.Compared with the protein database,this gene has no homologous protein and it is a typical orphan gene,we named it OsCPY.under normal conditions,We found the plants with down-regulated expression of OsCPY grew significantly faster than wild-type ZH11 at seedling stage,and under water flooding-stressed,the plants with down-regulated expression of OsCPY gene grew significantly faster than wild-type ZH11.In this study,we investigated the relationship between the expression of OsCPY and the rapid growth of rice seedlings and the primary molecular network regulation pathways.The main results were as follows:1.After germination of wild-type Zhonghua 11(ZH11),mutant(cpy),RNA interference(RNAi)and overexpressed(OE)and functional complementary(com),the growth of rice seedlings was observed.The results showed that in the 2 days,compared with ZH11,the aboveground and underground parts of cpy and RNAi plants increased by 0.50,0.85,0.25 and 1.3 times.The aboveground and underground parts of the OE plants were reduced by 0.41 times and 0.30 times.in the 3 days,compared with ZH11,the aboveground and underground parts of the cpy and RNAi plants increased by 0.53,0.61,0.41 and 0.52 times,and the aboveground and underground parts of the OE plants decreased by 0.38 and 0.74 times,There was no significant difference between com plants and ZH 11.Indicating that the expression of OsCPY was associated with the rapid growth of rice seedlings.2.To grow 3 days of the wild-type Zhonghuall(ZH11)and mutant(cpy)of the aboveground and underground part extracting RNA transcriptome sequencing analysis respectively,the results show that compared with ZH11,cpy aboveground parts of plants have raised expression of 563 genes,657 genes were expressed,underground part have raised 443 gene expression,gene expression by 1247.The functional annotation of these differentially expressed genes showed that they were mainly involved in transcriptional regulation,enzyme and regulation.We further analyzed the transcriptional regulation relationship between different genes in the aboveground and underground parts.In the aboveground part,there were 24 transcription factors related to the regulation of expression of 523 genes,while in the underground part,there were 11 transcription factors related to the regulation of transcription of 826 genes.3.In order to study the growth of the genetically modified materials under flooded conditions,the wild-type Zhonghua11(ZH11)and mutant(cpy),RNA interference(RNAi),overexpressed(OE)germination under flooded conditions,we found that the germination of 5 days,compared with ZH11,cpy,RNA interference plants of coleoptile increased 0.58 times and 0.47 times,while the coleoptile OE plants reduced 0.90 times,show that the expression of OsCPY and flooded rice seeds under the condition of rapid growth.4.Use online website PsRNATarget analysis,we can predict rice microRNA414 can controlled the expression of OsCPY,in order to verify whether there is a regulation relationship between the two,we builded microRNA414 knock out the transgenic plants,eventually won the expression of microRNA414 quantity significantly lower TO generation knockout plants CR-2 and no significant difference of CR-1 plant,and the above two kinds of genetically modified material in seedling growth were observed,in the 2 days and 3 days,The aboveground and underground parts of CR-2 plants were superior to the control group,while there was no significant difference between CR-1 plants and the control group.5.Using CPY as bait protein,we screened the candidate proteins interacting with CPY from the yeast two-hybrid library of rice seedlings,and finally obtained 15 candidate proteins interacting with CPY.The yeast two-hybrid point-to-point experiment verified that the proteins expressed by 0s02g0827500 interacted with CPY protein.6.Wild-type Zhonghua11(ZH11),mutant(cpy),RNA interference(RNAi)and overexpressed(OE)and functional complementary(com)were planted in a paddy field with a suitable growth environment after normal germination,and the plant height was measured after the rice plant was mature.The results showed that compared with ZH11,cpy plant height increased by 7.23%,RNA interference plant height increased by 7.33%,5.42%and 6.59%,and overexpressed plant height decreased by 11.70%,9.78%,10.53%and 9.78%,There was no significant difference between com plants and ZH11.