Identification and Characterization of an Efficient acyl-CoA:Diacylglycerol Acyltransferase 1 (\u3cem\u3eDGAT1\u3c/em\u3e) Gene from the Microalga \u3cem\u3eChlorella ellipsoidea\u3c/em\u3e

Background: Oil in the form of triacylglycerols (TAGs) is quantitatively the most important storage form of energy for eukaryotic cells. Diacylglycerol acyltransferase (DGAT) is considered the rate-limiting enzyme for TAG accumulation. Chlorella, a unicellular eukaryotic green alga, has attracted much attention as a potential feedstock for renewable energy production. However, the function of DGAT1 in Chlorella has not been reported. Results: A full-length cDNA encoding a putative diacylglycerol acyltransferase 1 (DGAT1, EC 2.3.1.20) was obtained from Chlorella ellipsoidea. The 2,142 bp open reading frame of this cDNA, designated CeDGAT1, encodes a protein of 713 amino acids showing no more than 40% identity with DGAT1s of higher plants. Transcript analysis showed that the expression level of CeDGAT1 markedly increased under nitrogen starvation, which led to significant triacylglycerol (TAG) accumulation. CeDGAT1 activity was confirmed in the yeast quadruple mutant strain H1246 by restoring its ability to produce TAG. Upon expression of CeDGAT1, the total fatty acid content in wild-type yeast (INVSc1) increased by 142%, significantly higher than that transformed with DGAT1s from higher plants, including even the oil crop soybean. The over-expression of CeDGAT1 under the NOS promoter in wild-type Arabidopsis thaliana and Brassica napus var. Westar significantly increased the oil content by 8–37% and 12–18% and the average 1,000-seed weight by 9–15% and 6–29%, respectively, but did not alter the fatty acid composition of the seed oil. The net increase in the 1,000-seed total lipid content was up to 25–50% in both transgenic Arabidopsis and B. napus. Conclusions: We identified a gene encoding DGAT1 in C. ellipsoidea and confirmed that it plays an important role in TAG accumulation. This is the first functional analysis of DGAT1 in Chlorella. This information is important for understanding lipid synthesis and accumulation in Chlorella and for genetic engineering to enhance oil production in microalgae and oil plants

Identification and characterization of an efficient acyl-CoA: diacylglycerol acyltransferase 1 (DGAT1) gene from the microalga Chlorella ellipsoidea

RT-PCR detection of DGAT1 genes in transgenic yeast (INVSc1). The yeast actin was used as an internal control. 1, The yeast transformed with pYES2.0; 2–5, the yeast expressing AtDGAT1, GmDGAT1, BnDGAT1 and CeDGAT1, respectively. (DOCX 55 kb

Measurements of NO2 and analysis of relationship between stratospheric NO2 and O3 over Zhongshan Station, Antarctica

We have already continually taken the measurement of total column amount of O3, NO2 as well as stratospheric O3, NO2 over Zhongshan Station, Antarctica. This paper analyzes the seasonal variation of NO2 and its relationship to O3 during the Antarctic Ozone Hole in 1993 - 1995 and points out that the decrease of atmospheric NO2 is one of the important reason to the strengthening of Antarctic Ozone Hole

Newly Identified Essential Amino Acids Affecting <i>Chlorella ellipsoidea</i> DGAT1 Function Revealed by Site-Directed Mutagenesis

Diacylglycerol acyltransferase (DGAT) is a rate-limiting enzyme in the synthesis of triacylglycerol (TAG), the most important form of energy storage in plants. Some residues have previously been proven to be crucial for DGAT1 activity. In this study, we used site-directed mutagenesis of the CeDGAT1 gene from Chlorella ellipsoidea to alter 16 amino acids to investigate effects on DGAT1 function. Of the 16 residues (L482R, E542R, Y553A, G577R, R579D, Y582R, R596D, H603D, H609D, A624R, F629R, S632A, W650R, A651R, Q658H, and P660R), we newly identified 5 (L482, R579, H603, A651, and P660) as being essential for DGAT1 function and 7 (E542, G577, R596, H609, A624, S632, and Q658) that significantly affect DGAT1 function to different degrees, as revealed by heterologous expression of the mutants in yeast strain INVSc1. Importantly, compared with CeDGAT1, expression of the mutant CeDGAT1Y553A significantly increased the total fatty acid and TAG contents of INVSc1. Comparison among CeDGAT1Y553A, GmDGAT1Y341A, AtDGAT1Y364A, BnDGAT1Y347A, and BoDGAT1Y352A, in which tyrosine at the position corresponding to the 553rd residue in CeDGAT1 is changed into alanine, indicated that the impact of changing Y to A at position 553 is specific for CeDGAT1. Overall, the results provide novel insight into the structure and function of DGAT1, as well as a mutant gene with high potential for lipid improvement in microalgae and plants

Calibration of Small-Grain Seed Parameters Based on a BP Neural Network: A Case Study with Red Clover Seeds

In order to enhance the accuracy of discrete element numerical simulations in the processing of small-seed particles, it is essential to calibrate the parameters of seeds within the discrete element software. This study employs a series of physical tests to obtain the physical and contact parameters of red clover seeds. A discrete element model of red clover seeds is established. Plackett–Burman Design, steepest ascent, and Central Composite Design experiments are sequentially conducted. The simulation deviation of the resting angle of red clover seeds is employed as the evaluation criterion for parameter optimization. The results indicate that the coefficients of static friction between red clover seeds, the coefficients of rolling friction between red clover seeds, and the coefficients of static friction between red clover seeds and the steel plates significantly influence the resting angle. Modeling was performed using a backpropagation neural network, a genetic algorithm–optimized BP network, particle swarm optimization, and simulated annealing. It was found that GA-BP ensured both accuracy and stability. Compared to the traditional response surface methodology, GA-BP showed better fitting performance. For the optimized red clover seed simulation, the error between the angle of repose and the physical experiment was 0.98%. This research provides new insights into the calibration of small-grain seed parameters, demonstrating the value of GA-BP for precision modeling

The Influence of the Air Cargo Network on the Regional Economy under the Impact of High-Speed Rail in China

There is little research on the impact of air cargo networks on regional economic development, which is especially notable considering that Chinese airlines gradually adjusted their networks after the introduction of high-speed rail (HSR). This empirical study aims to fill this research gap. Firstly, we used the Ordinary Least Squares (OLS) method to study the effect of the air cargo network on the regional economy. The results show that, in eastern and central China, the higher the clustering coefficient of the domestic air cargo network, the more significant their promotion effect becomes on the GDP per capita, with cities in eastern China benefitting the most from this effect. However, for super-scale cities, the clustering coefficient of the domestic air cargo network has a significant negative effect on the GDP per capita, which is likely because both the air and HSR passenger services crowd out the development opportunities for air cargo. Secondly, we applied the Difference-in-Difference (DID) method in order to measure the impact on the regional economy caused by air cargo under the impact of HSR. The results show that the aviation network adjusted for the impact of HSR produces heterogeneous effects on cities for different regions and scales, and that the international aviation network has greater impacts on cities than the domestic network. In eastern China, HSR and air cargo (both international and domestic networks) promote economic growth simultaneously; in central China, only domestic air cargo has a positive effect on the regional economy; in western China, neither HSR nor air cargo has an obvious effect on the regional economy. Policy implications&mdash;such as encouraging the cooperation of HSR and civil aviation&mdash;are discussed, and could help bring the functions of the air cargo network in regional economic development into full play