[Objective] The research aimed to construct the fusion protein expression vector of α-galactosidase-EGFP （enhanced green fluorescent protein） in cucumber controlled by CaMV35S promoter.[Method] CaMV35S promoter sequence and the coding region of EGFP were amplified by polymerase chain reactions （PCR） with vector pCambia 1303 as the template.Using reverse transcript PCR technology,with total RNAs of cucumber as template,the coding region of acid α-galactosidase Ⅰ in cucumber was amplified.The above three fragments were inserted into the multiple cloning sites of expression vector pCambia 1381c.The fusion expression vector of α-galactosidase-EGFP located at the C-terminal of the target genes was constructed.[Result] After enzyme digestion and sequencing,the fusion expression of α-galactosidase-EGFP in cucumber was constructed successfully.[Conclusion] The research laid the experimental basis for further study on the subcellular localization of α-galactosidase in cucumber.
The fruit yield of cucumber are associated with high input of nitrogen,which poses a risk of pollution to the environment.The EU-Rotate_N model has been used widely for its high performance in the simulation of vegetable growth,water and nitrogen dynamics.However,whether the underground water level affects the performance of the EU-Rotate_N model is unclear.In this study,we modified the groundwater level algorithms to the original model and named the modified model'the HG EU-Rotate_N model'.Experiments over two years on greenhouse cucumber with four different nitrogen(N)treatments(N1-N4)were conducted in Jiangsu Province,China,which has a high groundwater level(area_(HG)).Both original and modified models were used to simulate cucumber growth,water movement and N fate.For the soil water content,the measured values were significantly larger than the simulated values of the original model(value_(O))and closer to those of the HG model(value_(HG));for the soil available nitrogen concentration(SNC),the measured values were significantly higher and lower than value_(O) in 0−10 cm and 10−30 cm soil layers,respectively,and were also closer to those of the values_(HG).The higher SNC in the 0-10 cm soil layer indicated that a high groundwater level might increase the upwards movement of water and nitrogen in the 0-30 cm soil layer.The root mean square error,Nash Sutcliffe Efficiency and difference values show that the HG model was more applicable for areaHG than the original model.In this study,the nitrogen dosage of the N3 treatment was sufficient to meet the requirements of cucumber growth,indicating that the fertilization recommendation according to nitrogen nutrient balance was applicable in this area.
