The different effects of intramuscularly-injected lactate on white and brown adipose tissue in vivo

Molecular Biology Reports - Lactate is an important product of glycolysis metabolism during exercise and has long been recognized as an important metabolic signaling molecule involved in inhibiting...     

GenoBaits Soy40K: a highly flexible and low-cost SNP array for soybean studies

Science China Life Sciences -     

天山一号冰川表面冰尘和底部沉积层中可培养酵母菌系统发育类群的分布及生态生理特征

【目的】揭示乌鲁木齐河源天山1号冰川表面冰尘(CS)和底部沉积层(DS)可培养酵母菌系统发育类群及其结构组成差异,分析低温酵母菌代表菌株之间的生态、生理生化特性。【方法】利用4种培养基分离天山1号冰川可培养酵母菌,采用ITS基因序列分析确定菌种的系统进化地位。对分离菌株的最适生长温度、耐盐性和产酶等生态、生理学特性进行分析。【结果】从冰尘和底部沉积层中共分离出152株酵母菌菌株,通过ITSrRNA基因序列的NCBI比对和Rep-PCR指纹分型,结果表明酵母菌类群包括担子菌门(Basidiomycota)和子囊菌(Ascomycota),分属于14个属26种,其中担子菌门柄锈菌亚门(Pucciniomycotina)88株、伞菌亚门(Agariomycotina)24株,子囊菌门40株,冰川广布酵母菌Vishniacozyma victoriae为优势菌株(占比21.84%)。17种酵母的最适生长温度为15°C、2种为10°C、6种为20°C。25株代表酵母菌株产酶分析显示,产脂肪酶、淀粉酶、蛋白酶菌株分别为11株、11株、5株,6株3种酶都不产。【结论】天山1号冰川冰尘及底部沉积层可培养低温酵母系统发育类群结构存在差异,产低温酶活性高、稳定性好,为今后冰川低温酵母菌的研究提供有价值的数据支持。     

Front Cover Image,Volume 116, Number 11, November 2019

CRISPR/Cas9-guided cytidine deaminase enables C:G to T:A base editing in bacterial genome without introduction of lethal double-stranded DNA break, supplement of foreign DNA template, or dependence on inefficient homologous recombination. However, limited by genome-targeting scope, editing window, and base transition capability, the application of base editing in metabolic engineering has not been explored. Herein, four Cas9 variants accepting different protospacer adjacent motif (PAM) sequences were used to increase the genome-targeting scope of bacterial base editing. After a comprehensive evaluation, we demonstrated that PAM requirement of bacterial base editing can be relaxed from NGG to NG using the Cas9 variants, providing 3.9-fold more target loci for gene inactivation in Corynebacterium glutamicum. Truncated or extended guide RNAs were employed to expand the canonical 5-bp editing window to 7-bp. Bacterial adenine base editing was also achieved with Cas9 fused to adenosine deaminase. With these updates, base editing can serve as an enabling tool for fast metabolic engineering. To demonstrate its potential, base editing was used to deregulate feedback inhibition of aspartokinase via amino acid substitution for lysine overproduction. Finally, a user-friendly online tool named gBIG was provided for designing guide RNAs for base editing-mediated inactivation of given genes in any given sequenced genome ( www.ibiodesign.net/gBIG ).     

Two stochastic processes shape diverse senescence patterns in a single‐cell organism

Despite advances in aging research, a multitude of aging models, and empirical evidence for diverse senescence patterns, understanding of the biological processes that shape senescence is lacking. We show that senescence of an isogenic Escherichia coli bacterial population results from two stochastic processes. The first process is a random deterioration process within the cell, such as generated by random accumulation of damage. This primary process leads to an exponential increase in mortality early in life followed by a late age mortality plateau. The second process relates to the stochastic asymmetric transmission at cell fission of an unknown factor that influences mortality. This secondary process explains the difference between the classical mortality plateaus detected for young mothers’ offspring and the near nonsenescence of old mothers’ offspring as well as the lack of a mother–offspring correlation in age at death. We observed that lifespan is predominantly determined by underlying stochastic stage dynamics. Surprisingly, our findings support models developed for metazoans that base their arguments on stage‐specific actions of alleles to understand the evolution of senescence. We call for exploration of similar stochastic influences that shape aging patterns beyond simple organisms.     

Detoxification of insecticides,allechemicals and heavy metals by glutathione S‐transferase SlGSTE1 in the gut of Spodoptera litura

Insect glutathione S‐transferases (GSTs) play important roles in detoxifying toxic compounds and eliminating oxidative stress caused by these compounds. In this study, detoxification activity of the epsilon GST SlGSTE1 in Spodoptera litura was analyzed for several insecticides and heavy metals. SlGSTE1 was significantly up‐regulated by chlorpyrifos and xanthotoxin in the midgut of S. litura. The recombinant SlGSTE1 had V_max (reaction rate of the enzyme saturated with the substrate) and K_m (michaelis constant and equals to the substrate concentration at half of the maximum reaction rate of the enzyme) values of 27.95 ± 0.88 μmol/min/mg and 0.87 ± 0.028 mmol/L for glutathione, respectively, and V_max and K_m values of 22.96 ± 0.78 μmol/min/mg and 0.83 ± 0.106 mmol/L for 1‐chloro‐2,4‐dinitrobenzene, respectively. In vitro enzyme indirect activity assay showed that the recombinant SlGSTE1 possessed high binding activities to the insecticides chlorpyrifos, deltamethrin, malathion, phoxim and dichloro‐diphenyl‐trichloroethane (DDT). SlGSTE1 showed higher binding activity to toxic heavy metals cadmium, chromium and lead than copper and zinc that are required for insect normal growth. Western blot analysis showed that SlGSTE1 was induced in the gut of larvae fed with chlorpyrifos or cadmium. SlGSTE1 also showed high peroxidase activity. All the results together indicate that SlGSTE1 may play an important role in the gut of S. litura to protect the insect from the toxic effects of these compounds and heavy metals.     

DomeTree: a canonical toolkit for mitochondrial DNA analyses in domesticated animals

Mitochondrial DNA (mtDNA) is widely used in various genetic studies of domesticated animals. Many applications require comprehensive knowledge about the phylogeny of mtDNA variants. Herein, we provide the most up‐to‐date mtDNA phylogeny (i.e. haplogroup tree or matrilineal genealogy) and a standardized hierarchical haplogroup nomenclature system for domesticated cattle, dogs, goats, horses, pigs, sheep, yaks and chickens. These high‐resolution mtDNA haplogroup trees based on 1240 complete or near‐complete mtDNA genome sequences are available in open resource DomeTree ( http://www.dometree.org ). In addition, we offer the software MitoToolPy ( http://www.mitotool.org/mp.html ) to facilitate the mtDNA data analyses. We will continuously and regularly update DomeTree and MitoToolPy.     

基于细胞三维培养的气道平滑肌细胞收缩效应检测方法的建立与应用

气道平滑肌细胞(airway smooth muscle cells,ASMCs)是引起哮喘患者气道收缩狭窄、呼吸阻力增加的主要效应细胞。ASMCs收缩效应检测是研究哮喘病理生理机制、评估或研发新的支气管舒张药物的重要实验依据。而常用的活体组织张力检测以及单层培养ASMCs显微镜形态观察等方法存在样品取材或测量误差等问题。该研究利用胶原蛋白凝胶构建气道平滑肌细胞的三维立体培养模型,将凝胶与培养孔壁分离,在不同的时间点记录细胞收缩力作用下凝胶面积的变化值,以此反映ASMCs的收缩效应。结果显示,0.1,1,10 mmol/L的乙酰胆碱(acetylcholine,Ach)刺激后,凝胶面积均显著缩小,随着剂量增加ASMCs的收缩反应增强。提前加入肌球蛋白ATP酶抑制剂BDM(butanedione monoxime),能明显抑制Ach诱导的ASMCs收缩反应,说明该方法的可重复性和准确性好。     

Deep sequencing reveals a novel closterovirus associated with wild rose leaf rosette disease

A bizarre virus‐like symptom of a leaf rosette formed by dense small leaves on branches of wild roses (Rosa multiflora Thunb.), designated as ‘wild rose leaf rosette disease’ (WRLRD), was observed in China. To investigate the presumed causal virus, a wild rose sample affected by WRLRD was subjected to deep sequencing of small interfering RNAs (siRNAs) for a complete survey of the infecting viruses and viroids. The assembly of siRNAs led to the reconstruction of the complete genomes of three known viruses, namely Apple stem grooving virus (ASGV), Blackberry chlorotic ringspot virus (BCRV) and Prunus necrotic ringspot virus (PNRSV), and of a novel virus provisionally named ‘rose leaf rosette‐associated virus’ (RLRaV). Phylogenetic analysis clearly placed RLRaV alongside members of the genus Closterovirus, family Closteroviridae. Genome organization of RLRaV RNA (17 653 nucleotides) showed 13 open reading frames (ORFs), except ORF1 and the quintuple gene block, most of which showed no significant similarities with known viral proteins, but, instead, had detectable identities to fungal or bacterial proteins. Additional novel molecular features indicated that RLRaV seems to be the most complex virus among the known genus members. To our knowledge, this is the first report of WRLRD and its associated closterovirus, as well as two ilarviruses and one capilovirus, infecting wild roses. Our findings present novel information about the closterovirus and the aetiology of this rose disease which should facilitate its control. More importantly, the novel features of RLRaV help to clarify the molecular and evolutionary features of the closterovirus.