Analysis On Interrection Pathways Of ZmCLA2-2 Controlling Leaf Angle In Maize

Corn is not only an important food crop,but also an important grass crops and important industrial raw materials.With the decrease of cultivated land area and the increasing demand of maize,one of the most important ways to increase maize yield is to increase the density of maize.Leaf angle is one of the most important traits to determine the density of maize.Leaf angle development is the key trait of maize high-yielding breeding in current and future,It is important to study the key genes that influence the angle of corn leaf and construct the regulation network of key genes,which not only has important scientific value,but also has important significance for breeding new varieties with dense maize.In this study,the laboratory uses the compact inbred Yu 82 and the loose inbred sink Shen 137 as the material,cloning of candidate gene CLA2-2 by map-based cloning,this gene plays an important role in the transmission of brassinolide.The maize genetic transformation and fluorescence q RT-PCR technique proved that the gene regulated the leaf angle.On this basis,to further analyze the mechanism of action of the gene,in this study,yeast two-hybrid technique was used to screen proteins that interacted with CLA2-2,and the interaction genes of the leaves were further verified by bimolecular fluorescence complementary technique.The main results are as follows:1.According to the yeast two hybrid,Zm CLA2-2 c DNA was used as bait,a total of 38 positive clones interacted with Zm CLA2-2 were screened out of the Y82 yeast library,which has the correct reading frame ORF was 17.The bioinformatics analysis showed that: two b HLH transcription factors,two mitogen-activated protein kinasesmay be related to leaf angle development.The interaction between Zm CLA2-2 and b HLH transcription factors has been demonstrated in rice and Arabidopsis,to explore the new path of regulating corn leaf angle,The mitogen-activated protein kinase(MAPK)gene was subjected to bimolecular fluorescence complementation and rotary yeast validation.The results show that Zm CLA2-2 has a real interaction with candidate genes.2.MAPK was used as bait,a total of 36 positive clones interacted with MAPK were screened out of the Y82 yeast library,which has the correct reading frame ORF was 14,the bioinformatics analysis showed that: transcription initiation factor TFIID subunit A(EER4)is associated with regulation of cell division in cytokinin signaling pathways.Indicating that the gene may be related to leaf angle development.The gene was subjected to bimolecular fluorescence complementation and rotary yeast validation.The results show that MAPK has a real interaction with EER4.In summary,in the regulation of corn leaf angle formation of genes,brassin lactone signal transduction gene Zm CLA2-2 through MAPK pathway and cytokinin signal transduction gene EER interaction to control the size of leaf angle.That is,Zm CLA2-2 interacts with MAPK,MAPK interacts with EER4.Therefore,Zm CLA2-2 may control the size of corn leaf angle through MAPK,EER4.