Study On The Function Of Cunninghamia Lanceolate(Lamb.)Hook Transcription Factor ClMYB44

The MYB family(v-myb avian myeloblastosis viral oncogene homolog,short for MYB)is a class of transcription factors with highly conserved DNA binding domains,and has important biological functions in plant growth,development and abiotic stress responses.In addition,it's also an important factor affecting the regulation of lignin biosynthesis,and is generally involved in the plant metabolism control network.Phosphorus is a large number of elements necessary for plant growth and development.It is also one of the most important abiotic stresses faced by plants.Lack of phosphorus will cause traits such as short plants and yellow leaves.At the same time,phosphorus is also an important factor affecting lignin biosynthesis.Chinese fir(Cunninghamia lanceolata(Lamb.)Hook)is an important timber tree species in China,and an important part of China's artificial plantation forest.As its' excellent wood characteristics,it has a wide range of uses.However,the ClMYB444 transcription factor of Chinese fir has not yet been cloned,and its biological function in Chinese fir is remain unclear.Therefore,using the transcriptome data of C.lanceolata,this paper cloned the gene of C.lanceolata CIMYB44 and carried out the following research contents:1.The gene sequence of the transcription factor ClMYB44 was obtained by transcriptome sequence database.Specific primers with restriction sites were designed.Single-stranded cDNA was synthesized by reverse transcription of Chinese fir RNA as a template to amplify the gene of Chinese fir transcription factor ClMYB44.The full length of the gene is 1239bp,coded 413 aa.Bioinformation analysis result indicated that the secondary structure of ClMYB44 is a-helix and Random coil.2.We treated one-year old Cunninghamia lanceolata(Lamb.)Hook by using different concentration phosphorus for 15d,30d,45d,60d.The expression level of ClMYB44 in Chinese fir after phosphorus treatment was detected by qRT-PCR.It was found that ClMYB44 gene had the highest expression level at 8mg/L under different phosphorus concentrations,and too high or too low phosphorus concentration might inhibit the expression of ClMYB44 gene in Chinese fir.3.The pGBKT7-ClMYB44 yeast expression vector was constructed by enzyme digestion and ligation.The pGBKT7-ClA4YB44 was introduced into the activated yeast strain AH 109.The yeast one-hybrid experiment confirmed that ClMYB44 had transcriptional activity and confirmed that our cloned ClMYB44 was a transcription factor.4.The pBI121-ClMYB44 vector was constructed by enzymatic digestion and ligation method.The ClMYB44 transgenic poplar callus was obtained by Agrobacterium-mediated gene transformation method,and is inducing its rooting and seedling growth.Those results lay the foundation for further study on the function of ClMYB44in Chinese fir under phosphorus treatment,and provide technical support for molecular breeding and targeted variety improvement of Chinese fir.