Isolation And F Unctional Analysis Of Plant Architecture-related Genes TαOTUB1s In Triticum Aestivum L.

Wheat(Triticum aestivum L.)is one of the most important food crops in the world.With the increase of the world population and the decrease of the cultivated area,it is very important to increase the wheat yield.Identifition of the superior plant architecture-related genes will be valuable for molecular design breeding in wheat.In rice,the functional loss of the deubiquitinating enzyme OsOTUB1 leads to the accumulation of protein encoding OsIPA1/OsSPL14 that regulates of ideal plant type,resulting in fewer tillers,sturdier culms and larger panicles.To understand the molecular mechanism of plant type regulation of wheat,this study used bioinformatics methods to find and analyze the TaOTUBls gene,isolated the TaOTUBls gene and made a preliminary analysis of its function.The results are as followsThe bioinformatics technology was used to identify 50 members belonging to the Peptidase_C65 Otubain family in wheat.The conserved structural domain,gene structure,chromosome localization and gene expression analyses of the Peptidase C65 Otubain family were carried out in wheat,Further the TaOTUBls gene identified.The protein sequences of TaOTUBls and OTUB1s were compared,and it was found that OTUB1 proteins were highly conserved.TαOTUB1s have three alleles of TaOTUB1-A,TaOTUB1-B and TaOTUB1-D.The evolutionary tree analysis showed that TaOTUB1-A had a higher genetic relationship with OTUB1 in barley(Hordezun vulgare L.),TaOTUB1-A was choosed as the research object of this study.Transient expression results showed that TaOTUB1-A protein was located in the nucleus and cytoplasm.Quantitative PCR analysis results showed that TαOTUB1s was expressed in the shoot apexs at single and double-edged stages,axillary buds,spikelets and roots.In situ hybridization results show that TαOTUB1s is expressed in leaf primordium,young leaves,SAM and spikelets.Meanwhile,we analyzed the promoter cis-acting elements of TαOTUB1s,and found that TaOTUBls may be regulated by hormones and environmental factors.To study the effect of TaOTUBls on wheat plant architecture,we constructed pUbi TaOTUBl-RNAi vector.Further,the vector was transformed to the young embryos of Fielder wheat by agrobacterium-mediated method.Until now,we have obtained TaOTUB1-RNAi plants.Preliminary analysis results showed that tiller number of TaOTUB1-RNAi T2 generation 102#,405#and 513#lines was significantly decreased at seedling,jointing and heading stage compared with that of control Fielder plants.Meanwhile,Quantitative PCR analysis showed that the expression levels of TαOTUB1s were decreased while the expression of TaSPL17 was increased significantly in TaOTUB1-RNAi T2 linesThese results preliminarily indicate that TaOTUBls may be involved in the regulation of plant architecture in wheat.The conclusion of this study can provide some theoretical basis for investigating the mechanism of plant architecture regulation in wheat.