Effects of DHRS3 in C2C12 Myoblast Differentiation and Mouse Skeletal Muscle Injury

Myoblast differentiation is an essential process during skeletal muscle development. C2 C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro. Dehydrogenase/reductase(SDR family) member 3(DHRS3) is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol. Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation. However, the effect of DHRS3 on mouse muscle cell differentiation was unclear. The objective of current study was to determine if DHRS3 affected muscle cell differentiation, and if DHRS3 was involved in muscle regeneration. Protein expression was determined by western blot and immunofluorescence analysis. The activation and inhibition of DHRS3 increased and decreased C2 C12 myoblast differentiation respectively, which indicated that DHRS3 could affect C2 C12 myoblast differentiation. DHRS3 expression was significantly changed during muscle regeneration, with the regeneration of muscle injury, the expression of DHRS3 tended to increase first and then decrease. It suggested that DHRS3 might be involved in muscle regeneration. In summary, this study confirmed the involvement of DHRS3 in C2 C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development.     

基于GF-1和Sentinel-2时序数据的茶园识别

由于茶园大多分布在地形复杂的山区,地块破碎,分布零散,形状差异大、植被混杂且茶园所处环境长期受到云雨的影响,增加了茶园遥感识别的难度与不确定性,针对这一问题,该研究提出了利用高分1号(GF-1)和哨兵2号(Sentinel-2)时序数据提取茶园的方法,以浙江省武义县王宅镇为研究区,采用GF-1号为主要数据源,并利用MODIS地表反射率产品和Sentinel-2反射率数据,基于时空融合算法得到时间分辨率5 d的10 m Sentinel-2完整的时序数据。综合利用GF-1在空间细节方面的优势和重建的Sentinel-2高观测频率时序数据在反映茶树生长过程方面的优势,分别基于GF-1的光谱和纹理特征及GF-1的光谱、纹理特征和Sentinel-2时序特征两种特征组合方式,采用随机森林算法提取茶园。结果表明,GF-1光谱、纹理信息结合Sentinel-2时序信息分类结果的准确率、错误率、精确率、召回率和F1分数分别为96.91%、3.09%、89.00%、83.09%和0.86,仅基于GF-1光谱和纹理信息的分类准确率、错误率、精确率、召回率和F1分数分别为94.72%、5.28%、73.09%、84.61%和0.78,添加时序信息分类结果总体优于未添加时序信息的分类结果。表明高空间分辨率结合高频率时序遥感数据是提高茶园分类精度的有效手段。     

过表达ZmIBH1-1提高玉米苗期抗旱性

【目的】干旱是严重影响玉米生长发育进程的一个重要因素。挖掘玉米抗旱相关基因,通过转基因功能验证和转录组分析,解析关键基因在响应干旱胁迫过程中的分子调控机制,为抗旱分子育种和遗传改良提供理论依据。【方法】以玉米自交系B104（WT）为背景材料,利用农杆菌介导方法构建过表达ZmIBH1-1转基因株系（ZmIBH1-1-OE）;通过对转基因植株进行草铵膦抗性筛选、标记基因和目的基因PCR检测,以及运用实时荧光定量PCR检测目的基因的表达情况,鉴定阳性植株和株系;以WT和ZmIBH1-1-OE转基因株系为材料,通过干旱处理（20% PEG6000）,进行表型鉴定和耐旱生理生化指标测定,验证ZmIBH1-1的抗旱功能;通过对干旱胁迫下玉米4叶期转录组的比较分析,鉴定出差异表达的基因（differentially expressed genes,DEGs）;结合DAP-seq（DNA affinity purification sequencing）分析,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的下游靶基因,利用基因组可视化软件IGV（integrative genomics viewer）分析ZmIBH1-1蛋白结合候选靶基因的位置,然后通过Dual-Luciferase试验验证ZmIBH1-1蛋白与靶基因的调控关系。【结果】通过玉米遗传转化获得12个转化事件;T₃代中,能同时检测到标记基因Bar和目的基因ZmIBH1-1的植株有458个,实时荧光定量PCR检测结果表明,ZmIBH1-1-OE中ZmIBH1-1的表达量显著高于WT,株系3和株系8表达量最高,将其自交获得T₄代转基因株系用于后续试验。在干旱胁迫条件下,ZmIBH1-1-OE株系存活率、叶片相对含水量、叶绿素含量、可溶性蛋白含量及其生理生化指标（超氧化物歧化酶、过氧化物酶、过氧化氢酶活性）均显著高于WT,说明玉米中过量表达ZmIBH1-1赋予玉米更高的耐旱性。转录组分析结果表明,WT与ZmIBH1-1-OE株系在干旱胁迫下有1 214个差异表达基因;Gene Ontology（GO）功能富集分析结果表明,差异表达基因主要涉及生物过程、细胞组分和分子功能,如在生物过程中主要涉及到光合作用、应激响应、脱水响应等;KEGG富集分析表明,差异表达基因主要参与植物激素信号传导、新陈代谢等过程。结合转录组显著差异表达基因和DAP-Seq分析所得到ZmIBH1-1蛋白的靶基因,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的11个候选靶基因,包括2个钙信号相关基因、3个半胱氨酸代谢相关基因、1个bHLH转录因子、1个应激响应蛋白、1个谷胱甘肽转移酶、1个氧化还原过程蛋白和2个乙烯响应因子;基因组可视化结果显示ZmIBH1-1蛋白可以结合靶基因启动子区;随后通过Dual-Luciferase试验进一步表明,ZmIBH1-1蛋白可以直接作用于11个候选靶基因,其中,ZmIBH1-1蛋白可以促进ZmCa-M、ZmSYCO、ZmbHLH54、ZmGlu-r1、ZmCLPB3和ZmP450-99A2的表达,抑制ZmAGD12、ZmCYS、ZmCYSB、ZmERF-107和ZmEIN3的表达。此外,在干旱胁迫下NAC、WRKY、MYB等转录因子在ZmIBH1-1-OE和WT株系中也存在差异表达。【结论】ZmIBH1-1的过表达可以增强玉米苗期的耐旱性;ZmIBH1-1蛋白通过直接调控乙烯信号通路中的ZmERF-107和ZmEIN3的表达提高玉米的耐旱性;ZmIBH1-1蛋白通过直接调控钙信号相关基因ZmCa-M和ZmAGD12增强玉米的耐旱性;ZmIBH1-1蛋白可能通过间接调控NAC、WRKY、MYB等转录因子响应干旱胁迫。     

高产稳产大豆新品种兆丰2号的选育与应用

介绍了大豆新品种兆丰2号的选育过程、生物学特征特性、品质、抗性及产量表现,并总结了其配套栽培技术。     

CnAβ基因对大鼠RBL-2H3细胞脱颗粒的影响

过敏是人和养殖动物常见的疾病，而且发生的频率呈现逐渐增加的趋势。为了深入了解过敏反应的机制，本研究利用CRISPR/Cas9基因编辑技术，构建钙调磷酸酶A beta （calcineurin A beta，CnAβ）基因敲除的大鼠RBL-2H3细胞株，并探讨CnAβ基因对RBL-2H3细胞生长及脱颗粒的影响。选取大鼠CnAβ基因第一外显子为敲除靶点，设计并合成3个单导向RNA （single guide RNA，sgRNA），构建pX459-CnAβ-sgRNA质粒，并用脂质体3000将构建好的质粒转染到RBL-2H3细胞内；利用嘌呤霉素对转染细胞进行筛选，通过DNA测序验证获得CnAβ基因敲除的RBL-2H3细胞株；并检测CnAβ基因缺失对细胞增殖和脱颗粒的影响。结果表明，成功构建了CnAβ单基因敲除的大鼠RBL-2H3细胞株；CnAβ基因缺失对RBL-2H3细胞增殖、细胞的颗粒形成以及颗粒含量无显著影响，但显著抑制由细胞表面受体介导的RBL-2H3细胞脱颗粒作用。该研究结果有助于深入了解动物过敏性疾病的发生机制，为动物过敏性疾病的预防提供了理论基础。     

黄曲霉毒素B1、玉米赤霉烯酮和呕吐毒素的累加细胞毒性研究

由于饲料中多种霉菌毒素并存的几率比较高，本研究以仔猪肠上皮细胞（IPEC-J2）为模型，研究黄曲霉毒素B1（AFB1）、玉米赤霉烯酮（ZEA）和呕吐毒素（DON）的叠加细胞毒性。细胞毒性试验选用AFB1、ZEA和DON三种毒素作为响应面Box-Behnke设计的三个因素，以AFB1：10、20、30 μg/L，ZEA：150、300、450 μg/L，DON：500、1000、1500 μg/L作为Box-Behnke设计的三个编码水平。利用响应面设计构建得到17组复合霉菌毒素组合，以其对IPEC-J2细胞活力的影响作为参考指标，得到对细胞损伤程度最高和最低的霉菌毒素添加比例。结果表明：经方程预测后，得到细胞活力最低（霉菌毒素毒性最高）的AFB1、ZEA和DON组合为30、150 μg/L和1500 μg/L，经测定细胞活力为32.32%|得到细胞活力最高（霉菌毒素毒性最低）的AFB1、ZEA和DON组合为10、150 μg/L和600 μg/L，经测定细胞活力为53.01%。该结果为多种霉菌毒素叠加毒性的研究提供了依据。 [关键词] IPEC-J2细胞|黄曲霉毒素B1|玉米赤霉烯酮|呕吐毒素|细胞毒性     

Establishment and application of an accurate identification method for fragrant soybeans

In order to screen the aroma characteristics of soybean,a new method was established which can quickly quantify the content of 2-acetyl-1pyrroline (2-AP),an important compound related to soybean aroma,using gas chromatography-mass spectrometry (GC-MS).Based on peak profile,total peak area and retention time as test indexes,an accurate identification method for fragrant soybeans was established.The optimum parameters of the protocol consisted of column temperature70°C,sample injector temperature 180°C,optimum extraction alcohol content 1 m L,Na Cl content 0.1 g,ultrasonication time10 min,and extraction time 1 h,which were established by using the orthogonal test of single factors and three factors with four levels (L_9(3)~4).2-AP content of leaves had significant correlations with seeds,which were easier to measure.The protocol was simple and easy to carry out,consumed only small amounts of reagents,and provided accurate and reliable results with good reproducibility.A total of 101 soybean genotypes from different geographical sources were analyzed using this protocol.The results showed that the average content of 2-AP was 0.29 mg L~(–1),ranging from 0.094 to 1.816 mg L~(–1),and the genetic diversity index was 0.54.Among all genotypes-tested,they were classified into three grades,including seven elite genotypes identified as"grade one fragrant soybeans",which were Zhonglong 608,Heinong 88,Ha13-2958,Hongmiandou,Heinong 82,Huangmaodou,and Jiyu 21.These results provide both an identification technique and several elite aroma genotypes for gene discovery and good quality breeding in soybean.     

Evaluation on the fitness and population projection of Nilaparvata lugens in response to elevated CO2 using two-sex life table

Abstract

The life history and fitness of Nilaparvata lugens being reared under ambient level (current CO₂ concentration), medially elevated (550?µL/L) and highly elevated (750?µL/L) CO₂ concentration for long-term generation were compared using two-sex life table instead of traditional age-specific life table. The results showed that significantly longer larval duration and lower fecundity of N. lugens were observed in 750?µL/L relative to 550?µL/L treatment (P?<?0.05). Accordingly, 550?µL/L CO₂ significantly enhanced population parameters of N. lugens, including significantly higher intrinsic rate of increase (r), finite rate (λ) and net reproductive rate (R_O), but not for 750?µL/L CO₂. Taken together, N. lugens performs enhanced development rate, fecundity and survival in individual life history and higher potential in population multiplication under 550?µL/L CO₂ level, while only indicates the less enhanced development rate and survival without significant increased capacity of population expansion under 750?µL/L CO₂ level. These results should facilitate predicting the fitness and potential population damage of N. lugens, which is valuable for the integrated control of N. lugens in the future increasing CO₂ concentration.     

渠堤化城镇河岸带的植被特征及其用地影响

渠堤化是干扰城镇河流的常见建设方式，掌握植物特征及对其产生显著影响的用地因素，对于优化渠化的城镇河岸植被带具有重要意义。采用样带全面调查的方法，对北京段已基本全线渠堤化潮白河河岸带植物进行调查，通过多重比较分析河岸带植物丰富度在不同空间位置的差异，并利用NMDS非约束排序拟合解译的周边缓冲区土地利用数据，探讨对渠堤化河岸带植物丰富度和种类分布产生显著影响的因素。结果表明:1）河岸带植物种类174种（乔、灌、草分别为52、29、90种），以草本为主；2）全线渠堤化城镇河流河岸带植物群落种类及丰富度均存在建成区和非建成区的显著差异，总体植物、乔木和灌木植物丰富度在建成区显著大于非建成区，但入侵植物沿渠堤化河岸带不存在空间位置的差异，入侵可能沿渠堤蔓延；3）周边缓冲区道路、其他不透水地表面积是城镇地区全线渠堤化河岸带植物丰富度的显著正相关影响因子，对主要观赏灌木和乔木种类分布存在积极影响，对原生灌木和草本种类分布存在消极影响，这种分布特征受人工干扰影响。     

单端孢霉烯族毒素生物脱毒技术研究进展

单端孢霉烯族毒素在粮食和饲料中污染严重,主要是T-2毒素和脱氧雪腐镰刀烯醇(DON)等,其毒性作用强,对人和动物产生严重危害。因此,单端孢霉烯族毒素的脱毒方式逐渐成为研究热点,尤其是生物脱毒。很多微生物被证实能够降解霉菌毒素,包括细菌、真菌和酵母菌,可将毒素降解成低毒或无毒的产物。论文以生物脱毒为出发点,从细菌、真菌和酵母菌三个角度概述了单端孢霉烯族毒素中T-2毒素和DON的脱毒研究现状,以期为后续开发并挖掘更加安全高效的生物脱毒方式提供参考。