无机碳源对零价铁介导的自养脱氮序批式反应器处理高氨氮废水的影响

无机碳源作为自养微生物的能量来源,是影响自养脱氮细菌富集的重要因素。文章通过添加不同量的KHCO3作为ZVI介导的自养脱氮体系中无机碳源,研究反应体系的脱氮效果影响趋势以及适宜的无机碳源添加量。结果如下:1)KHCO3作为无机碳源可以显著提高NH^+4-N去除率,KHCO3添加量越多,其NH^+4-N去除率越高。KHCO3添加量分别为2 g,1 g,0.5 g的SBR反应器R2,R1,R0.5的NH^+4-N去除率分别为98.39%,65.18%,44.56%,相较不添加KHCO3的R0分别提高86.5%,53.29%,32.67%。ZVI的添加会降低KHCO3对NH^+4-N氧化的促进作用。2)KHCO3可明显提高TIN去除效果,促进总氮脱除的高低顺序是R1>R0.5>R2,SBR反应器R2,R1,R0.5的平均TIN去除率分别为19.01%,32.04%,27.62%,相较于空白组R0分别提高了10.60%,23.63%,19.21%。3)KHCO3可中和硝化反应产生的H^+,对反应体系具有缓冲作用,可使微生物处于较适宜的酸碱环境中,更有利于反应器的稳定运行。4)适量的无机碳源KHCO3可以促进氨氧化细菌和厌氧氨氧化细菌的活性。该研究探讨了无机碳源在零价铁脱氮体系中的影响趋势,为铁型脱氮技术提供了理论支撑。     

Cf-2/Rcr3pim番茄品系对南方根结线虫的抗性测定

Cf-2/Rcr3~(pim)基因型番茄不仅能够抵御番茄叶霉菌的侵染,而且对马铃薯金线虫的寄生也有一定的抑制效果。为挖掘根结线虫的新抗性资源,本研究采用室内人工接种法测定了Cf-0/Rcr3~(pim)、Cf-2/Rcr3-3和Cf-2/Rcr3~(pim)基因型番茄品系对南方根结线虫的抗感性。抗性评价结果显示,Cf-0/Rcr3~(pim)品系对南方根结线虫表现高感,Cf-2/Rcr3-3品系为中感,而Cf-2/Rcr3~(pim)品系则为感病。与Cf-0/Rcr3~(pim)和Cf-2/Rcr3-3基因型相比,Cf-2/Rcr3~(pim)基因型番茄品系虽然对南方根结线虫侵染的敏感性略低,但是不能阻止线虫在根系上的大量繁殖,不适于根结线虫的防控应用。     

ROCK promotes high glucose-induced cardiomyocyte apoptosis by inhi-biting PI3K/Akt signaling pathway

AIMTo investigate whether Rho-associated coiled-coil kinase (ROCK) is involved in high glucose-induced apoptosis of primary cardiomyocytes by regulating PI3K/Akt signaling pathway. METHODSPrimary Wistar rat cardiomyocytes were cultured and identified by α-sarcomeric actin (α-SCA) immunohistochemistry. Cardiomyocytes were treated with 5.5, 33 and 40 mmol/L glucose for 48 h. The cell viability was measured by MTT assay, and the mRNA expression of ROCK1 and ROCK2 in the cardiomyocytes was detected by RT-qPCR. Flow cytometry was used to analyze the apoptosis of the cardiomyocytes. The protein levels of ROCK1, ROCK2, cleaved caspase-3, Bcl-2, PI3K, Akt and p-Akt were determined by Western blot. In order to confirm the regulatory effect of ROCKs on PI3K/Akt signaling pathway, the cells were divided into control group (5.5 mmol/L glucose), high glucose group (33 mmol/L glucose) and high glucose+Y27632 (ROCK inhibitor) group. Western blot was used to detect the protein levels of ROCK1, ROCK2, PI3K, Akt and p-Akt. RESULTSAfter 48 h of high glucose exposure, the values of relative cell viability in 33 and 40 mmol/L glucose groups were (79.71±2.43)% and (68.41±7.49)%, respectively, both of which were significantly decreased compared with normal control group (P<0.05). After 48 h of high glucose exposure, the relative mRNA levels of ROCK1 and ROCK2 in 33 and 40 mmol/L glucose groups were significantly increased compared with normal control group (P<0.05). Compared with normal control group, the apoptotic rate in 33 and 40 mmol/L glucose groups was increased significantly (P<0.05). Compared with normal control group, the protein expression of ROCK1, ROCK2 and cleaved caspase-3 in 33 and 40 mmol/L glucose groups was increased (P<0.05), while the protein expression of Bcl-2 was decreased (P<0.05). No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed, while the protein level of p-Akt in 33 and 40 mmol/L glucose groups was decreased compared with normal control group (P<0.05). Compared with high glucose group, the expression of ROCK1 and ROCK2 was decreased in high glucose+Y27632 group. No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed. Compared with normal control group, the protein level of p-Akt in high glucose group was decreased, and the protein level of p-Akt in high glucose+Y27632 group was increased significantly compared with high glucose group. CONCLUSION Under high glucose environment, ROCK may reduce the level of p-Akt by inhibiting the PI3K/Akt signaling pathway， thus promoting the apoptosis of cardiomyocytes.     

利用CRISPR/Cas9系统敲除葡萄中VviEDR2提高对白粉病的抗性

利用CRISPR/Cas9系统定点编辑葡萄白粉病感病基因VviEDR2（Enhanced disease resistance 2），在VviEDR2的DUF1336结构域设计靶位点VviEDR2-T1，构建CRISPR/Cas9敲除载体，通过农杆菌介导法转化‘无核白’葡萄胚性愈伤组织。对PCR阳性植株进行靶位点扩增测序，结果表明，共有8个转基因植株在靶位点处发生不同类型的双等位基因突变，编辑效率为32%；突变体植株生长势较弱，叶片较小，茎秆丛生、细弱。进一步对突变体植株进行抗病检测，结果表明，接种葡萄白粉菌（Erysiphe necator Schw.）5 d后，突变体植株叶片上白粉菌孢子仅能萌发出少量较短初级菌丝，表皮细胞产生大量明显的H2O2，而野生型叶片中白粉菌萌发出大量初级菌丝、次级菌丝和吸器，无明显H2O2产生。这些结果表明，可以利用CRISPR/Cas9技术编辑葡萄感病基因VviEDR2，提高葡萄白粉菌抗性。     

新型缓控释增效肥研究进展和发展前景

传统肥料面临着利用率低、产能过剩等问题,新型缓控释肥通过新型包膜材料或者纳米技术,实现养分释放速度的可控性;通过添加信号物质,实现植物和化肥的互动,将植物养分需求规律作为主导因素,实现肥料的按需供给,达到减少化肥用量,提高肥料利用效率的目的。从定义、研发背景、分类及发展趋势等方面介绍了新型缓控释肥的研究进展,并对其未来发展进行了展望。     

不同植被措施下排土场边坡细沟发育时空特征

为明确不同恢复年限植被措施对煤矿排土场边坡水土流失的防治效应,采用样方调查法,以内蒙古准格尔旗永利煤矿矿区排土场边坡为研究对象,以裸露边坡(CK)为对照,研究不同恢复年限(1a、3a、5 a)的沙柳方格+沙棘+沙打旺(SHA)和沙柳方格+沙打旺(SA)2种措施对细沟发育的影响。结果表明:(1)1a时CK、SHA与SA措施细沟宽度均集中分布在4～8 cm,细沟深度均集中分布在2～4 cm;3 a时CK、SHA与SA措施细沟宽度则集中分布在4～8 cm、8～12 cm、4～8 cm,细沟深度均集中分布在4～6 cm;5 a时CK与SHA措施细沟宽度均集中分布在8～12 cm,而细沟深度则集中分布在4～6 cm和8～14 cm;(2)CK(1～5 a)、SHA措施(1～5 a)和SA措施(1～3 a)的细沟平均宽分别为7.57～11.35 cm、7.58～13.31 cm和5.57～6.14 cm,细沟平均深分别为3.38～6.23 cm、4.19～10.34 cm、2.59～4.24 cm,三者的细沟平均宽深比分别为2.39、2.12和2.05,平均细沟密度分别为1.52～5.25 m·m–2、1.42～1.68 m·m–2和1.88～2.25 m·m–2;(3)1 a时CK、SHA和SA措施的细沟宽深比随坡长变化幅度较大,随着恢复年限增加,宽深比则呈下降趋势,CK、SHA措施和SA措施的细沟密度和细沟侵蚀量均随坡长增加呈增大趋势;(4)与CK相比,1 a时SHA和SA措施边坡细沟侵蚀模数分别减小25.0%和25.86%,两种措施减蚀效果差别不大,而3 a时则分别减小了61.73%和35.31%,SHA措施减蚀效果显著增强。研究结果可以为矿区排土场边坡的植被合理布设提供科学依据与理论指导。     

Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.     

Hormonal changes play important roles in the key period of superior and inferior earshoot differentiation in maize

The upper earshoots with higher superiority usually have higher yield potential and higher efficiency. To determine the key period for the asynchronous differentiation of superior and inferior earshoots and how hormones are involved in this process, a two-year experiment was designed using two maize hybrids: Suyu 41 (S41, single-ear hybrid) and AN 101 (A101, double-ear hybrid). The results showed that the lag of lower earshoot differentiation was not only caused by the delay of the differentiation starting time but also related to extension of the duration in spikelet differentiation (stage II) and sexual organ formation stage (stage IV). From 12 days before silking (DBS), the contents of indole-3-acetic acid (IAA), zeatin riboside (ZR)+zeatin (ZT), and gibberellic acid (GA₃) in both upper and lower earshoots of the two hybrids increased dramatically and then decreased quickly. ABA slightly increased in the two hybrids and then decreased slowly in S41, while it was maintained at a high level in A101. At 8 DBS, i.e., the transition period from floret differentiation to sexual organ formation stage, not only the growth of upper-to-lower earshoot difference (ULED), but also the values for ULED of IAA, ZR+ZT and GA₃ were all significantly higher in S41 than in A101. Furthermore, the upper-to-lower hormone ratios IAA_U//AA_L and (ZR+ZT)_U/(ZR+ZT)_L were also much higher in the single-ear hybrid than in the double-ear hybrid, while the GA_3U/GA_3L and ABA_U/ABA_L had no significant differences. In addition, the time course of ULED_hormone/ULED_{earshoot growth rate} also suggested that the hormones work in different ways in earshoot superiority/inferiority formation. The delayed differentiation of lower ear shoots was conclusively related to the later initiation of differentiation and the longer durations of specific differentiation stages. Compared with the regulating roles of IAA and ZR+ZT in the key period (8 DBS) of superiority/inferiority differentiation, GA₃ seems to be affected earlier, while ABA contributes little to this process.     

‘72杨’韧皮部的构造与理化性能

将‘72杨’、杉木和毛竹的木质部进行对比试验,观察并测试‘72杨’韧皮部的解剖构造以及理化特性,为其高值化利用提供基础数据。使用场发射环境扫描电镜、X射线衍射仪、傅里叶变换红外光谱仪等设备和NREL标准对‘72杨’韧皮部的微观结构、结晶度、化学成分等物化性质进行测定与分析。研究结果表明,‘72杨’韧皮部中韧皮薄壁细胞和筛管分子占细胞总面积的(81.9±1.8)%,结构相对简单。‘72杨’韧皮部气干密度为0.358 g/cm3,绝干密度为0.321 g/cm3;而木质部的相应密度较高,分别为0.497和0.482 g/cm3;‘72杨’韧皮部结晶度仅为19.4%,比木质部低8.7%。‘72杨’韧皮部纤维素、半纤维素、木质素的含量分别为28.7%,11.1%,24.1%,均低于木质部中相应成分的含量,且木质化程度低,半纤维素以木糖为主。此外,由红外谱图发现‘72杨’韧皮部含有单宁、酚类、胼胝质等物质。‘72杨’韧皮部具有低密多孔、结构疏松、结晶度低、木质素含量低、抽提物含量高等特点。因此,‘72杨’韧皮部特别有利于机械(能耗低)或化学(抗降解屏障低)降解以及物化改性(多孔、可及性强),可提取酚类、单宁、胼胝质等物质用于工业应用,研究结果可为‘72杨’韧皮部的高值化利用提供重要的理论依据。     

利用CRISPR/Cas9技术编辑BADH2-1/BADH2-2创制香米味道玉米新种质

【目的】香味是作物的重要食味品质之一。2-乙酰-1-吡咯啉（2-acetyl-1-pyrroline,2-AP）是主要香味物。BADH2是控制水稻等作物香味性状的关键基因,敲除该基因可以产生香味稻米。利用CRISPR/Cas9基因编辑技术在北京市农林科学院自育的玉米骨干亲本京724上敲除BADH2同源基因,以期获得有香米味道的玉米新种质材料。【方法】利用Ensembl数据库在线BLAST工具,将水稻OsBADH2蛋白序列在拟南芥、水稻和玉米蛋白序列数据库中进行序列比对,获得上述3个物种的BADH基因家族成员,并利用UniProt蛋白数据库中的结构域信息进行验证。进一步使用MEGA软件进行系统进化分析,获得玉米BADH2同源基因作为候选编辑靶标。基于CRISPR/Cas9基因编辑技术的原理,在候选基因的外显子处设计特异性靶点,并构建入CRISPR/Cas9基因编辑载体。再以玉米自交系京724为受体,利用农杆菌介导的遗传转化方法,通过磷酸甘露糖异构酶基因（phosphomannose isomerase,PMI）抗性筛选获得阳性转基因植株。转基因株系经测序明确其在靶基因中产生的突变类型。利用气相色谱质谱联用仪（gas chromatography-mass spectrometry,GC-MS）检测基因编辑株系T₁籽粒中香米主要香味物质2-AP的含量,以确认京724在基因编辑前后2-AP含量的变化。【结果】系统进化分析发现,玉米中存在2个BADH2同源基因,分别命名为ZmBADH2-1和ZmBADH2-2。ZmBADH2-1位于第4染色体,ZmBADH2-2位于第1染色体。2个基因均包含15个外显子和14个内含子,第4外显子间的核苷酸序列高度同源。在2个基因的第4外显子区域设计靶点并构建入CRISPR/Cas9基因编辑载体,通过遗传转化获得28株转基因株系。PCR扩增及测序分析结果显示,其中10株材料的2个ZmBADH2s在靶点区域均发生突变,突变基因型包括双等位突变和多等位突变,突变类型为不同数量的碱基缺失和插入。质谱检测结果显示玉米ZmBADH2双基因突变体籽粒中存在与香稻同样成分的2-AP。随机选取的4个T₁代基因编辑株系籽粒中,2-AP平均含量分别为438.29、404.63、348.65和161.82 μg·kg^-1,而未经过编辑的京724中未检测到2-AP。【结论】利用CRISPR/Cas9技术对玉米ZmBADH2-1和ZmBADH2-2同时进行定点敲除,创制出籽粒中具有香米味道的玉米骨干亲本新种质材料。