Preparation of the pure diastereomeric forms of S- (5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2- cyclopentene and their evaluation as irreversible inhibitors of S-adenosylmethionine decarboxylase from Escherichia coli

The conformationally restricted S-adenosylmethionine analogue AdoMac (S-(5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2-cyclopentene has been shown to act as an enzyme activated, irreversible inhibitor of theEscherichia coli form of the enzyme S-adenosylmethionine decarboxylase. Inactivation of the enzyme is presumably initiated by formation of an imine linkage between the inhibitor and the terminal pyruvate of the enzyme, followed by base-catalyzed elimination of methylthioadenosine and generation of a latent electrophile. Removal of the driving force for the elimination of methylthioadenosine resulted in a reversibly binding inhibitor. Thus, the thioether analogue corresponding to AdoMac, and the corresponding dihydro derivative (H₂-AdoMac), reversibly inhibit the enzyme. AdoMac was resolved into its four pure diastereomeric forms, and each diastereomer was evaluated as an irreversible inhibitor of the enzyme. The K_I values for the individual diastereomers range between 3.83 and 39.6 μM, with the cis-1S,4R diastereomer being the most potent inhibitor. However, the k_inact values for the four diastereomers are not significantly different, suggesting that the binding of each diastereomer to the enzyme is configuration-dependent, while the subsequent inactivation likely proceeds through a single intermediate which is formed from each of the four diastereomers. Since each pure diastereomer represents a distinct conformational mimic exhibiting restricted sidechain rotation, the data suggests that these and related analogues may be useful as conformational probes for the catalytic site of AdoMet-DC.     

辽宁省爬行动物名录及地理区划更新

基于此前辽宁省爬行动物调查资料和分类学研究,结合近年来野外调查进展,对辽宁省爬行动物名录和区系进行修订,并报道辽宁省蛇类新记录二种——乌梢蛇（Ptyasdhumnades）和黑头剑蛇（Sibynophischinensis）。根据从辽宁省凌源市野外获得的影像资料进一步确认黑眉锦蛇（Elaphe taeniura）在辽宁省的分布。另依据采集自葫芦岛市的1号王锦蛇（E. carinata）标本对该种在辽宁省的分布予以补充描述。截至2023年5月,辽宁省记录爬行动物2目11科21属37种,其中,龟鳖目3科4属5种,有鳞目蜥蜴亚目3科5属10种,蛇亚目5科12属22种。爬行动物中,国家一级重点保护野生动物3种,国家二级重点保护野生动物6种。此外,有15个物种被《中国生物多样性红色名录脊椎动物第三卷爬行动物》评估为受威胁物种。根据辽宁省爬行动物最新分布信息,将辽宁省划分为6个动物地理省,分别为辽东山地丘陵省、辽东半岛山地丘陵省、辽河平原省、努鲁儿虎山北麓丘陵台地及西辽河沙地省、辽西山地丘陵省和冀辽山地省,其中,冀辽山地省为本研究中新增动物地理省。扩大原辽东半岛丘陵省涵盖范围,并将其名称修订为“辽...     

Reduced Application of Nitrogen Fertilizer Affects the Carbon Metabolism of Leaves and Maintains the Number of Flowers in Coreopsis tinctoria

This study examined the effects of nitrogen (N) fertilizer reduction on the carbon (C) metabolism and yield of Coreopsis tinctoria. A two-year (2020–2021) hydroponic experiment was conducted in accordance with a randomized complete group design with five N levels [0.875 mM Ca(NO₃)₂ (N1), 1.750 mM Ca(NO₃)₂ (N2), 3.500 mM Ca(NO₃)₂ (N3), 7.000 mM Ca(NO₃)₂ (N4), and 14.000 mM Ca(NO₃)₂ (N5)] and three replications. The results showed that low N significantly affected the functional leaf weight, C metabolism, and flower bud (or flower) numbers of C. tinctoria at harvest. Lower-N levels, especially those of the N2 treatment, significantly increased Rubisco, sucrose synthase (SS), sucrose phosphate synthase (SPS), soluble acid invertase (SAI), glucose 6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) activity and maintained the flower number of C. tinctoria. In addition, the balance of carbohydrates (sucrose, starch, glucose, and fructose) and ATP contents was more efficiently maintained under relatively low-N levels. These findings might suggest that reduced application of N fertilizer affects the C metabolism of leaves and maintains the number of flowers in Coreopsis tinctoria. Applying relatively low-N fertilizer levels is also a promising cultivation strategy for C. tinctoria.

     

金银花花形基因的发掘及生物信息学分析

金银花作为我国重要的中药材,具有消炎、抗菌、抗病毒、抗氧化、防癌等多种功效。随着金银花市场供需矛盾日益加剧,通过分子标记辅助选择育种方法来培育高产优质品种势在必行。通过NCBI的Blast工具扫描金银花蛋白组数据发掘花形候选基因,并执行候选基因的亲缘关系分析、结构域分析、表达模式分析、理化性质分析、蛋白质结构预测等一系列生物信息学分析。依据拟南芥调控花形的ABE类基因,通过NCBI-Blast工具扫描金银花氨基酸序列,筛选出包含MADS结构域的8个调控花形的金银花候选基因。经LjaFGD表达模式分析发现,金银花的花中GWHGAAZE016592和GWHGAAZE014905表达量显著高于其他部位,可能正向调控金银花花形。GWHGAAZE014905是一个包含MADS结构域的调控花器官发育的B类基因;GWHGAAZE016592是AP3同源基因。生物信息学分析发现,GWHGAAZE016592和GWHGAAZE014905均是稳定的亲水蛋白,属于非分泌蛋白,包括Motif1、Motif3、Motif4、Motif2、Motif6和Motif5,蛋白质三级结构模板为6byy.2.A和4ox0.2.C。GWHGAAZE014905被定位到细胞核上,而GWHGAAZE016592被定位到叶绿体上,且包含1个位于151~173 bp的跨膜螺旋区域,属于膜蛋白。研究结果为分子标记辅助选择方式培育道地高产优质金银花品种提供了基因资源和分子标记。     

Soil amendment with biochar and manure alters wood stake decomposition and fungal community composition

Biochar and manure can be used for sustainable land management. However, little is known about how soil amendments might affect surface and belowground microbial processes and subsequent wood decomposition. In a split-split-split plot design, we amended soil with two rates of manure (whole plot; 0 and 9 Mg ha⁻¹) and biochar (split plot; 0 and 10 Mg ha⁻¹). Wood stakes of three species (hybrid poplar, triploid Populus tomentosa Carr.; aspen, Populus tremuloides Michx.; and pine, Pinus taeda L.) were placed in two positions (horizontally on the soil surface, and inserted vertically in the mineral soil), which served as a substrate for fungal growth. In 3 years, the decomposition rate (density loss), moisture content, and fungal community (via high-throughput sequencing methods) of stakes were evaluated. Results indicated that biochar and/or manure increased the wood stake decomposition rates, moisture content, and operational taxonomic unit abundance. However, the richness and diversity of fungi were dependent on wood stake position (surface > mineral), species (pine > the two Populus), and sample dates. This study highlights that soil amendment with biochar and/or manure can alter the fungal community, which in turn can enhance an important soil process (i.e., decomposition).     

A short-chain carbonyl reductase mutant is an efficient catalyst in the production of (R)-1,3-butanediol

R-1,3-butanediol (R-1,3-BDO) is an important chiral intermediate of penem and carbapenem synthesis. Among the different synthesis methods to obtain pure enantiomer R-1,3-BDO, oxidation–reduction cascades catalysed by enzymes are promising strategies for its production. Dehydrogenases have been used for the reduction step, but the enantio-selectivity is not high enough for further organic synthesis efforts. Here, a short-chain carbonyl reductase (LnRCR) was evaluated for the reduction step and developed via protein engineering. After docking result analysis with the substrate 4-hydroxy-2-butanone (4H2B), residues were selected for virtual mutagenesis, their substrate-binding energies were compared, and four sites were selected for saturation mutagenesis. High-throughput screening helped identify a Ser154Lys mutant which increased the catalytic efficiency by 115% compared to the parent enzyme. Computer-aided simulations indicated that after single residue replacement, movements in two flexible areas (VTDPAF and SVGFANK) facilitated the volumetric compression of the 4H2B-binding pocket. The number of hydrogen bonds between the stabilized 4H2B-binding pocket of the mutant enzyme and substrate was higher (from four to six) than the wild-type enzyme, while the substrate-binding energy was decreased (from −17.0 kJ/mol to −29.1 kJ/mol). Consequently, the catalytic efficiency increased by approximately 115% and enantio-selectivity increased from 95% to 99%. Our findings indicate that compact and stable substrate-binding pockets are critical for enzyme catalysis. Lastly, the utilization of a microbe expressing the Ser154Lys mutant enzyme was proven to be a robust process to conduct the oxidation–reduction cascade at larger scales.     

Correlation between fibronectin and its receptor in chick myoblast differentiation

Alterations in the amount of fibronectin and in the number of its receptors during myoblast differentiation of chicken embryo were investigated. The amount of fibronectin in the cell surface pool as measured by immunoblotting decreased during myogenesis To identify and characterize the fibronectin receptors on the myoblasts, the interactions of the 28,000 dalton (28 kDa) amino terminal fragment and 85,000 dalton (85 kDa) cell-binding fragment of fibronectin with my-oblasts were examined. The binding of the 28 kDa fragment was found to be time-dependent and reached a maximum level within 60 min. The unlabeled 28 kDa fragment inhibited the binding of the radioiodinated 28 kDa fragment, whereas the unlabeled 85 kDa fragment and antibody to integrin did not inhibit it, suggesting that the 28 kDa fragment interacts with the matrix assembly receptors but not with the cell adhesion receptors. There was a single class of 3.4 × 10⁵ binding sites per cell with an apparent dissociation constant of 1.4 × 10^?7 M on 30 hr old myoblasts. The specific binding of the radioiodinated 28 kDa fragment to myoblasts decreased as the fusion proceeded. This decrease of binding was consistent with the decrease in the amount of fibronectin. Furthermore, the levels of fibronectin and binding of the radioiodinated 28 kDa fragment in the fusion-blocked myoblasts by EGTA treatment appeared to remain constant. These results suggest that the decrease and/or loss of fibronectin during myoblast fusion is closely correlated with the alteration of fibronectin receptors and with the fusion of myoblasts.     

kar交配法转移YACs至新宿主的研究

利用改进的kar交配法,将一个含有340kb人基因组DNA的YAC片段的供体酵母菌株YAC23与受体菌株YLB504进行交配,以选择平板对所形成的候选YAC导入菌进行筛选。经PCR分析,候选YAC导入菌在404bp处有一个扩增带,即具有受体菌株的交配型(MAT α)。进一步用脉冲电泳进行核型鉴定,证实YACs己成功地进入受体,实现了YACs从一个宿主到另一个宿主之间的转移。     

人工栽培条件下三叶木通座果及果实生长特性研究

用栽培三叶木通为试材,研究了其座果及果实生长的特性。结果表明：人工栽培条件下,三叶木通座果（雌花）率可以提高到１３．５％,是野生三叶木通的２ １００％;以先年第１、２次攀援茎第３￣３０节位结果为主;３月底开花,花期３０ｄ左右。花后第２０￣５０ｄ（４月中旬至５月上旬）花序、雌花及幼果（子房）大量脱落,出现明显的脱落高峰。三叶木通果实纵向生长呈双Ｓ曲线布,生长期１５０ｄ左右,可划分成５个时期：受精结实