高、低氮浓度对2株真眼点藻的生长和油脂积累的影响

【目的】研究氮浓度对真眼点藻纲(Eustigmatophyceae)的2株高产油微藻大真眼点藻(Eustigmatos magnus,EM)和波氏真眼点藻(Eustigmatos polyphem,EP)的细胞形态、生长、总脂含量、脂质组成和脂肪酸组成与含量的时序变化规律。【方法】利用高氮(18.0 mmol/L NO3?-N)和低氮(3.6 mmol/L NO3?-N)浓度培养微藻。【结果】形态观察结果表明,大真眼点藻(E. magnus)和波氏真眼点藻(E. polyphem)营养细胞具有1个周生的裂叶状叶绿体,细胞质中有液泡,内含能够振动的颗粒物,以及一个较为明显的红色色素体;生殖方式通过形成2个D形或4个四角形的似亲孢子;随着培养周期的延伸和营养盐的消耗,细胞中油体逐步形成,其数量不断增加,体积不断增大。实验结果表明,初始氮浓度对2种微藻的总脂积累及生长均有显著影响(P<0.05),低氮浓度下2种微藻的生物质浓度分别为9.0 g/L和8.5 g/L,均低于高氮浓度下的生物质浓度。而低氮浓度下2种微藻的总脂、中性脂和总脂肪酸的含量以及总脂、中性脂与总脂肪酸的单位体积产率均明显高于高氮浓度组,其最高值分别为：59.10%、51.90%、46.95%和0.28、0.24、0.22 g/(L·d) (EM);64.20%、56.80%、50.01%和0.32、0.28、0.25 g/(L·d) (EP)。脂肪酸分析结果表明,两种微藻的脂肪酸主要成分均为棕榈酸(C16:0)、棕榈油酸(C16:1)、油酸(C18:1)和二十碳五烯酸(C20:5,EPA),四者的总含量(占总脂肪酸)分别达到85.83%和85.48%,其中棕榈油酸的含量最高。【结论】低氮浓度胁迫有利于大真眼点藻和波氏真眼点藻细胞内油脂的积累,两种微藻均为适合于生产生物柴油的油脂生产藻株。     

Optogenetic manipulation of cyclic guanosine monophosphate to probe phosphodiesterase activities in megakaryocytes

Cyclic guanosine monophosphate (cGMP) signalling plays a fundamental role in many cell types, including platelets. cGMP has been implicated in platelet formation, but mechanistic detail about its spatio-temporal regulation in megakaryocytes (MKs) is lacking. Optogenetics is a technique which allows spatio-temporal manipulation of molecular events in living cells or organisms. We took advantage of this method and expressed a photo-activated guanylyl cyclase, Blastocladiella emersonii Cyclase opsin (BeCyclop), after viral-mediated gene transfer in bone marrow (BM)-derived MKs to precisely light-modulate cGMP levels. BeCyclop-MKs showed a significantly increased cGMP concentration after illumination, which was strongly dependent on phosphodiesterase (PDE) 5 activity. This finding was corroborated by real-time imaging of cGMP signals which revealed that pharmacological PDE5 inhibition also potentiated nitric oxide-triggered cGMP generation in BM MKs. In summary, we established for the first-time optogenetics in primary MKs and show that PDE5 is the predominant PDE regulating cGMP levels in MKs. These findings also demonstrate that optogenetics allows for the precise manipulation of MK biology.     

锰矿区大叶胡枝子灌丛土壤的丛枝菌根真菌多样性

大叶胡枝子(Lespedeza davidii)为一种豆科灌木,具有较强耐受重金属胁迫及固氮能力,也是矿区一种重要修复植物。丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)是一类植物共生真菌,它在矿区与植物共生可促进植物生长、提高植物的抗逆性;但其种类、分布及其影响机制尚不够清楚。以大叶胡枝子灌丛土壤AMF群落为研究对象,采集矿区与非矿区大叶胡枝子冠下及其相邻空旷地土壤进行内部转录间隔区(ITS)测序,分析土壤理化性质,揭示不同样地土壤AMF群落的多样性及其影响因素。研究结果表明:(1)土壤样本共得到2961个可操作分类单元(OTU),其中属于AMF的OTU为66个;Silva数据库比对发现AMF有7目10科16属24种。AMF总体物种丰度为矿区相邻空旷地土壤>矿区冠下土壤>非矿区冠下土壤>非矿区相邻空旷地土壤,Shannon指数和Simpson指数与该趋势保持一致。(2)矿区土壤AMF群落以内养囊霉属(Entrophospora)等为优势属,非矿区冠下以斗管囊霉属(Funneliformis)等为优势属。稀有内养囊霉(Entrophospora infrequens)是4个样地共有的优势种,该种为广谱生态型种;Archaeospora sp.和Paraglomerales sp.为矿区冠下的优势种,它们对锰胁迫有较强的耐受性;Funneliformis sp.为非矿区冠下的优势种。(3)冗余分析(RDA)表明,AMF的种类和数量受到锰含量、土壤pH值以及全磷含量的显著影响。本研究结果说明AMF种类及多样性受到土壤理化性质的影响;矿区一些耐性AMF的存在提高了AMF多样性,它们有利于逆境条件下植物的生长。     

Structural and dynamic basis of DNA capture and translocation by mitochondrial Twinkle helicase

Twinkle is a mitochondrial replicative helicase which can self-load onto and unwind mitochondrial DNA. Nearly 60 mutations on Twinkle have been linked to human mitochondrial diseases. Using cryo-electron microscopy (cryo-EM) and high-speed atomic force microscopy (HS-AFM), we obtained the atomic-resolution structure of a vertebrate Twinkle homolog with DNA and captured in real-time how Twinkle is self-loaded onto DNA. Our data highlight the important role of the non-catalytic N-terminal domain of Twinkle. The N-terminal domain directly contacts the C-terminal helicase domain, and the contact interface is a hotspot for disease-related mutations. Mutations at the interface destabilize Twinkle hexamer and reduce helicase activity. With HS-AFM, we observed that a highly dynamic Twinkle domain, which is likely to be the N-terminal domain, can protrude ∼5 nm to transiently capture nearby DNA and initialize Twinkle loading onto DNA. Moreover, structural analysis and subunit doping experiments suggest that Twinkle hydrolyzes ATP stochastically, which is distinct from related helicases from bacteriophages.