The Cloning And Genetic Transformation Of Photoperiod Controlling Gene ZmSOC1 In Maize

Tropical and subtropical maize germplasm from the diversity center of corn contains abundant hereditary variation, which can broaden the genetic base of the temperate zone maize breeding. However, the photoperiod sensitivity severely of different tropical and subtropical maize germplasm limits the use of them in temperate zone on a large scale. Therefore, it is theoretical and practical importance to use the tropical and subtropical maize germplasm and regulate photoperiod sensitivity of photoperiod response of maize further, isolating and cloning the photoperiod regulator, study the function and expression profile in the photoperiod response.We cloned the ZmSOCl gene through orthologus gene cloning in maize inbreeding line 18-599, S37,271-1 and 274-1, which was the ortholog of SOC1 gene of Arabidopsis; We constructed the over expression vectors pCAMBIA1302-SOCl ox of ZmSOCl from maize inbreeding line 18-599 and 274-1 respectively, transformed them into the Arabidopsis through Floral Dip method, and obtained several transformants respectively; We constructed the over expression vectors pCAMBIA1302-SOCl ox and RNAi vector pJawohl8-SOC7 RNAi of ZmSOCl (inserted fragment is from maize inbreeding line 18-599), and transformed them into maize embryogenic callus of inbreeding line 18-599 by bombardment, and obtained several transformants of pCAMBIA1302-,SOC1 ox; It was checked the reliability of the operation of bombardment method and efficiency of gold grain coated with the plasmid 35S..GUS which transient expressed in the maize embryogenic callus after bombardment In addition, we approached the inductivity of embryogenic callus of different sizes of immature embryo, the selection of different embryogenic callus with different concentration of Basta, and the survival of regeneration plant of transgenic. The main results are as follows:1 According to the method of operation of particle gun in the materials and methods, the GUS gene was imported to the maize embryogenic callus, and expressed in high efficiency after the plasmid of 35S::GUS bombarded the maize embryogenic callus.2 Three different sizes of immature embryo of inbreeding line 18-599,0.5-0.8 mm, 0.8-1.2 mm and 1.2-2.0 mm induced embryogenic callus.96%was the best inductivity when the size of the immature embryo was 0.8-1.2 mm.3 Different maize inbreeding line 18-599(W),18-599(R), and 416051 adapted to different concentration of Basta. It was indicated that best selection concentration of the three inbreeding line is 2-3 mg/L,10 mg/L and 2-3 mg/L, respectively.4 We constructed the over expression vector pCAMBIA13O2-SOCl ox and loss-of-function expression vector pJawohl8-SOC1 RNAi of ZmSOCl successfully. The PCR result indicated the over expression vector had been transformed into Arabidopsis successfully. The over expression vector and loss-of-function expression vector were also transformed into embryogenic callus of maize inbreeding line 18-599. The PCR dectection and sequencing indicated that only the over expression vector was transformed into maize embryogenic callus successfully.