Functional Analysis Of Maize Gene,ZmLBD16 Affecting Lateral Organ Differentiation In Arabidopsis Thaliana

LBD(Lateral Organ Boundaries Domain)gene family at the N-terminus has a conserved LOB domain,is a higher plant-specific transcription factor.The gene family is involved in the development of plant organ boundaries,affecting callus formation and is affected by plant hormones and the environment.In this study,a long 880 bp sequence was obtained from maize by homologous cloning,encoding 261 amino acids,and the typical class I LBD gene family was named Zm LBD16.The overexpression vector of this gene was constructed and transferred into the Arabidopsis mustard.On the basis of this study,the expression characteristics of Zm LBD16 in maize were analyzed.The phenotypic determination of transgenic Arabidopsis thaliana was carried out.The expression of Zm LBD16 transgenic Arabidopsis thaliana in different tissues and different developmental stages was analyzed.Stress and hormone treatment of transgenic Arabidopsis thaliana,Zm LBD16 gene expression patterns were analyzed,this study to further reveal the Zm LBD16 gene in maize lay the foundation for the function.The main results of this study are as follows:1.The results showed that the expression of ZmLBD16 gene was in the order of root> stem> leaf in the inbred line,and the expression of Zm LBD16 gene in the inbred line was analyzed by fluorescence quantitative RT-PCR analysis.The expression of Zm LBD16 gene in N04 was as follows: stem> root> leaf.2.Zm LBD16 gene was transformed into Arabidopsis thaliana,the leaves of transgenic plants were curled down,the leaves were smaller,the plant height was decreased,and the plant height was significantly different from that of wild type Arabidopsis.The length of transgenic Arabidopsis thaliana was small,The difference between grain weight and 1000-grain weight was significant.The transgenic Arabidopsis thaliana and wild-type seedlings were cultured vertically,The difference between the root length and the root number of the transgenic Arabidopsis thaliana was significantly different.Zm LBD16 gene affects the growth and development of roots,stems,leavesand embryos of Arabidopsis thaliana and has a great influence on the growth of lateral roots.3.The expression of Zm LBD16 gene was increased first and then increased again at the 7th week,and the expression level of Zm LBD16 gene was the highest at the 7th week,followed by the 5th week,week.RT-PCR analysis of Zm LBD16 transgenic Arabidopsis thaliana showed that the expression of Zm LBD16 gene in different tissues was as follows: stem> leaf> fruit> flower.4.The leaves of Zm LBD16 transgenic Arabidopsis thaliana were cultured on B5 medium containing 2,4-D.When 2,4-D concentration was 0.15 mg / L,transgenic Arabidopsis leaves could form callus and wild Type of Arabidopsis thaliana leaves can not form callus,2,4-D concentration increased transgenic Arabidopsis leaves formation of callus growth status better.Thus,this suggests that maize LBD homologous gene Zm LBD16 affects callus formation and may function similarly to Arabidopsis thaliana LBD16 gene.5.The results showed that the expression of Zm LBD16 gene was increased and the growth status of transgenic Arabidopsis thaliana was better than that of wild type when Na Cl concentration was higher than 0.15 M.The germination rate of wild type Arabidopsis thaliana was 80 ~ 90% when mannitol concentration was 0.15 M,and the germination rate of Zm LBD16 transgenic Arabidopsis was only 40 ~ 50%,and the germination rate of Zm LBD16 transgenic Arabidopsis thaliana was 80 ~ The The expression of Zm LBD16 gene was increased and the seedlings were dry and killed in the transgenic seedlings at 40 ? for 3h.6.The results showed that when the concentration of 6-BA was 0.05 M,the expression of Zm LBD16 gene was higher than that of untreated control,and the expression level of Zm LBD16 gene was lower than that of 0M The results showed that the gene was regulated by 6-BA.The transgenic Arabidopsis thaliana was treated with four 2,4-D concentrations.The expression of Zm LBD16 gene was higher than that of CK at 0.15 M,indicating that 2,4-D and Zm LBD16 gene was negatively regulated.With different concentrations of IAA,the expression of Zm LBD16 gene was higher than that of control at the concentration of less than 0.1M.When the concentration was more than 0.15 M,the expression level was lower than that of untreated.The results showed that low concentration of IAA promoted Zm LBD16 gene The expression of Zm LBD16 gene was significantly different from that of control at different concentrations of NAA,and the expression of Zm LBD16 gene was significantlyhigher than that of control at the concentration of 0.10.15 M,and the concentration of Zm LBD16 was higher than that of control at other concentrations The expression level was lower than that of the control,indicating that the gene was regulated by NAA.