ABSTRACT
Sweet potato is an important food crop that can also be used as an industrial raw material. Sucrose is the main form of long-distance carbohydrate transport in plants, and sucrose transporter (SUT) regulates the transmembrane transport and distribution of sucrose during plant growth and metabolism. Moreover, SUT plays a key role in phloem mediated source-to-sink sucrose transport and physiological activities, supplying sucrose for the sink tissues. In this study, the full-length cDNA sequences of IbSUT62788 and IbSUT81616 were obtained by rapid amplification of cDNA ends (RACE) cloning according to the transcripts of the two SUT coding genes which were differentially expressed in sweet potato storage roots with different starch properties. Phylogenetic analysis was performed to clarify the classification of IbSUT62788 and IbSUT81616. The subcellular localization of IbSUT62788 and IbSUT81616 was determined by transient expression in Nicotiana benthamiana. The function of IbSUT62788 and IbSUT81616 in sucrose and hexose absorption and transport was identified using yeast functional complementarity system. The expression pattern of IbSUT62788 and IbSUT81616 in sweet potato organs were analyzed by real-time fluorescence quantitative PCR (RT-qPCR). Arabidopsis plants heterologous expressing IbSUT62788 and IbSUT81616 genes were obtained using floral dip method. The differences in starch and sugar contents between transgenic and wild-type Arabidopsis were compared. The results showed IbSUT62788 and IbSUT81616 encoded SUT proteins with a length of 505 and 521 amino acids, respectively, and both proteins belonged to the SUT1 subfamily. IbSUT62788 and IbSUT81616 were located in the cell membrane and were able to transport sucrose, glucose and fructose in the yeast system. In addition, IbSUT62788 was also able to transport mannose. The expression of IbSUT62788 was higher in leaves, lateral branches and main stems, and the expression of IbSUT81616 was higher in lateral branches, stems and storage roots. After IbSUT62788 and IbSUT81616 were heterologously expressed in Arabidopsis, the plants grew normally, but the biomass increased. The heterologous expression of IbSUT62788 increased the soluble sugar content, leaf size and 1 000-seed weight of Arabidopsis plants. Heterologous expression of IbSUT81616 increased starch accumulation in leaves and root tips and 1 000-seed weight of seeds, but decreased soluble sugar content. The results obtained in this study showed that IbSUT62788 and IbSUT81616 might be important genes regulating sucrose and sugar content traits in sweet potato. They might carry out physiological functions on cell membrane, such as transmembrane transport of sucrose, sucrose into and out of sink tissue, as well as transport and unloading of sucrose into phloem. The changes in traits result from their heterologous expression in Arabidopsis indicates their potential in improving the yield of other plants or crops. The results obtained in this study provide important information for revealing the functions of IbSUT62788 and IbSUT81616 in starch and glucose metabolism and formation mechanism of important quality traits in sweet potato.
Subject(s)
Ipomoea batatas/metabolism , Arabidopsis/metabolism , Sucrose/metabolism , Saccharomyces cerevisiae/metabolism , DNA, Complementary , Phylogeny , Plants, Genetically Modified/genetics , Membrane Transport Proteins/metabolism , Starch/metabolism , Plant Proteins/metabolism , Gene Expression Regulation, PlantABSTRACT
Objective To explore the teaching effect of acute poisoning course among students majoring in rescue medicine. Methods From March to June 2013, 30 students majoring in rescue medicine were randomly divided into two groups for the clinical practice. Case-based learning was used in experimental group and cases was tracked from one department to another in chronological order or in reverse order to explore the cause of poisoning and the development of disease and to ob-tain the complete medical records of patients. Traditional rotating internship was used in control group. Poisoning cases in the department were taken as the starting point and end point and the medical records of patients were obtained. The clinical teaching effect was compared between the two groups through questionnaire survey and final examination. The data were calculated by SAS 9.2 software, mean comparison between the two groups was conducted by t test and enumeration data were analyzed by chi-square test. P0.05),however, theory examination results were increased significantly in this year compared with those of previous year((88.1±6.0) vs. (79.3±12.8), P<0.01). Questionnaire survey demonstrated that teachers and students' satisfaction were 95.3%(143/150) and 93.3%(140/150) respectively regarding the ten advantages of the track-ing type internship. Conclusions Tracking type internship with poisoning case-based learning is a good combination of lecture-based learning and problem-based learning. It is helpful to cultivate and exercise students' ability to apply the knowledge.