微生物肥料及其生产应用中的问题

微生物肥料和微生物制剂是新世纪实行绿色农业的重要技术保障,关键是因其中含有大量的有效微生物的生命活动产生特定的肥效或其他生理功能,导致增产,微生物肥料包括根瘤菌肥,解磷菌肥、解钾菌肥、5406菌肥,植物根际促生菌,VA菌根等等,有机肥料堆制剂中含有多种降解农业有机废料的菌种,可缩短堆肥周期,提高养份利用率,由于微生物本身的特殊性,相关市场监督机制不健全,微生物肥料的生产,应用领域存在很多问题,甚至出现明显的伪科学,科研、生产、行政各有关部门应加强基础研究,提高全民科普水平,健全市场监督机制,依靠专家设计严格试验,优选可靠技术,保证良好的经济效益和社会效益。     

不同土地利用类型对丹江口库区土壤氮矿化的影响

氮(N)素是陆地生态系统净初级生产力的重要限制因子, 土地利用类型的变化对生态系统氮循环过程有着重要的影响。采用PVC顶盖埋管原位培养的方法, 对丹江口库区清塘河流域相邻的侧柏(Platycladus orientalis)人工林、人工种植灌木林地和农田3种土地利用类型的氮素矿化和硝化作用进行了研究。结果表明, 侧柏人工林、灌木林地和农田的NH₄⁺-N浓度(mg·kg^-1)依次为1.33 ± 0.20、1.67 ± 0.17和1.62 ± 0.13, 不同土地利用类型间的NH₄⁺-N浓度无显著性差异; 而3种土地利用类型下土壤NO₃^--N浓度(mg·kg^-1)差异显著, 农田NO₃^--N浓度(9.00 ± 0.73)显著高于侧柏人工林(1.27 ± 0.18)和灌木林地(3.51 ± 0.11)。NO₃^--N在灌木林地和农田中分别占土壤无机氮库的67.8%和84.8%, 是土壤无机氮库的主要存在形式; 而侧柏人工林中NO₃^--N和NH₄⁺-N浓度则基本相等。土壤硝化速率(mg·kg^-1·30 d^-1)从农田(7.13 ± 2.19)、灌木林地(2.56 ± 1.07)到侧柏人工林(0.85 ± 0.10)显著性降低。侧柏人工林、灌木林地和农田的矿化速率(mg·kg^-1·30 d^-1)依次为0.98 ± 0.12、2.52 ± 1.25和6.58 ± 2.29。矿化速率和硝化速率显著正相关, 但是矿化速率在不同的土地利用类型间差异不显著。培养过程中灌木林地和农田NH₄⁺-N的消耗大于积累, 氨化速率为负值, 导致灌木林地和农田矿化速率小于硝化速率。氮素的矿化和硝化作用受土壤含水量和土壤温度的影响, 并对土壤含水量更为敏感。土壤C:N与土壤矿化和硝化速率显著负相关。研究结果表明: 土地利用类型的变化会改变土壤微环境和土壤C:N, 进而会影响到土壤氮循环过程。     

木本植物非结构性碳水化合物变化及其影响因子研究进展

非结构性碳水化合物(NSC)是植物新陈代谢过程中重要的供能物质,NSC库的动态变化不仅反映了植物体内的碳收支状况,还影响着植物的生理代谢活动.为了预测树木甚至整个森林生态系统对未来气候变化的响应和适应能力,本文综述了树木NSC研究的最新进展,重点介绍了NSC库的季节和区域性变化,分析总结了影响树木NSC含量和分配格局变化的主要环境因素.最后还对未来气候变化背景下树木NSC库可能的响应策略和适应状况进行了讨论,展望了未来树木NSC领域的研究方向.     

Nonenzymatic chromophore attachment in biliproteins: conformational control by the detergent Triton X-100

While chromophore attachment to alpha-subunits of cyanobacterial biliproteins has been studied in some detail, little is known about this process in beta-subunits. The ones of phycoerythrocyanin and C-phycocyanin each carry two phycocyanobilin (PCB) chromophores covalently attached to cysteins beta84 and beta155. The differential nonenzymatic reconstitution of PCB to the apoproteins, PecA, PecB, CpcA and CpcB, as well as to mutant proteins of the beta-subunits lacking either one of the two binding cysteins, was studied using overexpression of the respective genes. PCB adds selectively to Cys-84 of CpcA, CpcB, PecA, and PecB, but the bound chromophore has a nonnative configuration, and in the case of CpcA, is partly oxidized to mesobiliverdin (MBV). The oxidation is independent of thiols but can be suppressed by ascorbate. The addition to Cys-beta84 is suppressed in the presence of detergents like Triton X-100, in favor of an addition to Cys-beta155 yielding the correctly bound chromophore. Triton X-100 also inhibits oxidation of the chromophore during addition to CpcA. The effect of Triton X-100 was studied on the isolated components of the reconstitution system. Absorption, fluorescence and circular dichroism spectra indicate a major conformational change of the chromophore upon addition of the detergent, which probably controls the site selectivity of the addition reaction, and inhibits the oxidation of PCB to MBV.     

Pluripotency-associated miR-290/302 family of microRNAs promote the dismantling of naive pluripotency

The molecular mechanism controlling the dismantling of naive pluripotency is poorly understood. Here we show that microRNAs (miRNAs) have important roles during naive to primed pluripotency transition. Dgcr8^−/− embryonic stem cells (ESCs) failed to completely silence the naive pluripotency program, as well as to establish the primed pluripotency program during differentiation. miRNA profiling revealed that expression levels of a large number of miRNAs changed dynamically and rapidly during naive to primed pluripotency transition. Furthermore, a miRNA screen identified numerous miRNAs promoting naive to primed pluripotency transition. Unexpectedly, multiple miRNAs from miR-290 and miR-302 clusters, previously shown as pluripotency-promoting miRNAs, demonstrated the strongest effects in silencing naive pluripotency. Knockout of both miR-290 and miR-302 clusters but not either alone blocked the silencing of naive pluripotency program. Mechanistically, the miR-290/302 family of miRNAs may facilitate the exit of naive pluripotency in part by promoting the activity of MEK pathway and through directly repressing Akt1. Our study reveals miRNAs as an important class of regulators potentiating ESCs to transition from naive to primed pluripotency, and uncovers context-dependent functions of the miR-290/302 family of miRNAs at different developmental stages.     

水牛CD14 cDNA的克隆、表达及鉴定

目的:克隆水牛白细胞分化抗原14(buffalo cluster of differentiation antigen14,bCD14)基因,表达bCD14蛋白,并进行Western Blot鉴定.方法:采用RT-PCR方法从水牛外周血白细胞中扩增bCD14基因,构建重组质粒pET28a-bCD14,转化入E coli BL21,IPTG诱导表达,对表达蛋白进行可溶性分析及Western blot鉴定.结果:bCD14基因含有一个1 122bp的开放阅读框,编码373个氨基酸;与印度水牛、挪威大鼠和人CD14的cDNA序列同源性分别为97.95%、68.78%、78.60%,氨基酸同源性分别为96.78%、61.27%、72.34%;主要以包涵体形式表达,表达蛋白经Western Blot鉴定,得到了一条约46 kD的特异性条带.结论:该文成功克隆了bCD14基因,表达了bCD14蛋白,为进一步揭示水牛抵抗革兰氏阴性菌感染的免疫机制奠定了基础.     

Cyclosporin A通过MEK/ERK1/2信号通路调节滋养细胞titin表达

探讨MEK/ERK1/2信号通路在Cyclosporin A(CsA)诱导滋养细胞表达titin中的作用。应用RT-PCR、Western blot检测CsA诱导的滋养细胞titin的表达水平,Western blot检测CsA作用于滋养细胞后ERK1/2的活化程度,并观察MEK特异性抑制剂U0126对其mRNA转录的影响。发现CsA以时间和剂量依赖方式诱导titin表达,并刺激滋养细胞ERK1/2的活化,U0126以剂量依赖方式抑制CsA诱导的titin表达。结果表明CsA通过活化MEK/ERK1/2信号通路诱导滋养细胞titin 的表达,改变其生物学行为,从而有利于胚胎着床及早期发育。     

大豆C3HC4型RING锌指蛋白基因GmRZFP1克隆与表达分析

锌指蛋白在调节植物防卫基因表达和抗性反应上起关键作用。目前,对大豆中C3HC4型RING锌指蛋白基因的研究不多。本研究利用核蛋白筛选系统(NTT)筛选大豆(铁丰8号)干旱处理5h的cDNA文库,获得一个RING锌指蛋白基因。该基因全长927bp,编码308个氨基酸,含有C3HC4-type RING锌指结构域,命名为GmRZFP1。系统进化树分析显示,Gm-RZFP1属于C3HC4-type锌指亚家族。Real-time PCR结果表明,GmRZFP1基因受干旱、高盐、高温、低温、乙烯和ABA等胁迫诱导表达,表明该蛋白涉及多种胁迫相关的信号传导途径。亚细胞定位结果表明,163hGFP-GmRZFP1融合蛋白定位于细胞核中。本研究结果有助于研究该类基因在大豆逆境应答反应中的作用,阐明大豆抗逆分子机制。     

Metabolism in 1,3‐propanediol fed‐batch fermentation by a D‐lactate deficient mutant of Klebsiella pneumoniae

Klebsiella pneumoniae HR526, a new isolated 1,3‐propanediol (1,3‐PD) producer, exhibited great productivity. However, the accumulation of lactate in the late‐exponential phase remained an obstacle of 1,3‐PD industrial scale production. Hereby, mutants lacking D ‐lactate pathway were constructed by knocking out the ldhA gene encoding fermentative D ‐lactate dehydrogenase (LDH) of HR526. The mutant K. pneumoniae LDH526 with the lowest LDH activity was studied in aerobic fed‐batch fermentation. In experiments using pure glycerol as feedstock, the 1,3‐PD concentrations, conversion, and productivity increased from 95.39 g L^?1, 0.48 and 1.98 g L^?1 h^?1 to 102. 06 g L^?1, 0.52 mol mol^?1 and 2.13 g L^?1 h^?1, respectively. The diol (1,3‐PD and 2,3‐butanediol) conversion increased from 0.55 mol mol^?1 to a maximum of 0.65 mol mol^?1. Lactate would not accumulate until 1,3‐PD exceeded 84 g L^?1, and the final lactate concentration decreased dramatically from more than 40 g L^?1 to <3 g L^?1. Enzymic measurements showed LDH activity decreased by 89–98% during fed‐batch fermentation, and other related enzyme activities were not affected. NADH/NAD⁺ enhanced more than 50% in the late‐exponential phase as the D ‐lactate pathway was cut off, which might be the main reason for the change of final metabolites concentrations. The ability to utilize crude glycerol from biodiesel process and great genetic stability demonstrated that K. pnemoniae LDH526 was valuable for 1,3‐PD industrial production. Biotechnol. Bioeng. 2009; 104: 965–972. © 2009 Wiley Periodicals, Inc.     

马来酸氟伏沙明片联合氨磺必利治疗精神分裂症患者的疗效及安全性分析

目的:分析马来酸氟伏沙明片联合氨磺必利治疗精神分裂症患者的临床疗效及安全性。方法:选择我院2014年2月~2018年2月收治的182例精神分裂症患者,按随机数字表法分为对照组99例和研究组83例。对照组采用氨磺必利治疗,研究组在对照组基础上联合马来酸氟伏沙明片治疗。比较两组临床疗效,治疗前后血脂代谢、总胆汁酸(TBA)水平,阳性与阴性症状量表(PANSS)评分,生活质量,及不良发生情况。结果:治疗后,研究组总有效率为91.57%,显著高于对照组(P<0.05);两组治疗前后空腹血糖(FPG)、糖化血红蛋白(HbA1c)、甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白-C(LDL-C)、高密度脂蛋白-C(HLD-C)、总胆汁酸(TBA)水平比较均无统计学意义(P>0.05);两组治疗后PANSS评分均较治疗前显著下降,生活质量评分较治疗前均明显上升,且研究组PANSS评分显著低于对照组,而生活质量评分明显高于对照组,差异均有统计学意义(P<0.05)。治疗过程中,两组均有体重增加、口渴及便秘发生,两组不良反应发生情况比较差异无统计学意义(P>0.05)。结论:马来酸氟伏沙明片联合氨磺必利治疗能够提高精神分裂症患者疗效,对机体糖脂代谢及肝功能影响较小,有良好的用药安全性。