大白菜NHX基因家族的鉴定与表达分析

Na+/H+逆向转运蛋白(Na+/H+antiporter,NHX)基因家族在植物响应盐胁迫中发挥重要作用。本研究鉴定了大白菜NHX基因家族成员,并分析了大白菜NHX基因(Brassica rapa ssp.Pekinensis NHX,BrNHXs)响应高温、低温、干旱和盐胁迫等非生物逆境的表达模式。结果表明,在大白菜中共鉴定到9个NHX基因家族成员,分布在大白菜的6条染色体上,其氨基酸数目在513–1154 aa之间,相对分子量集中在56804.22–127856.66 kDa,等电点位于5.35–7.68之间。该基因家族成员主要存在于液泡中,基因结构完整,外显子的数目介于11–22之间。大白菜NHX基因家族编码的蛋白质二级结构都具有α-螺旋、β-转角和不规则卷曲结构,其中α-螺旋发生频率较高。实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)分析显示,该基因家族成员在高温、低温、干旱和盐胁迫下均有不同程度地响应,且在不同时间表达差异显著。以BrNHX02和BrNHX09对这4种胁迫的响应最为显著,表达量在处理72 h时均显著上调,可作为候选基因进一步验证其功能。     

攀西高原不同轮作制度下土壤固氮微生物nifH基因多样性与群落结构特征

[背景]关于高原生境轮作制度对土壤固氮微生物群落组成及多样性的影响研究尚少。[目的]深入认识攀西高原不同轮作制度对农田土壤肥力及土壤固氮微生物nifH基因群落结构与多样性的影响,以期建立合理的轮作制度。[方法]以凉山州冕宁县不同作物轮作制度[包括光叶紫花苕-烤烟(分轮作15年和20年两种,分别为G1和G2)、苦荞-烤烟(KQ)、大麦-烤烟(DM)和撂荒(CK)]的土壤为研究对象,通过化学分析和Illumina MiSeq技术,对土壤理化性质、土壤固氮微生物nifH基因多样性及群落组成进行分析。[结果]撂荒土壤全氮、铵态氮、硝态氮、有机碳和含水量最显著(P<0.05)。KQ轮作下土壤有效磷和速效钾分别提高了43.0%和2.60%,而DM轮作下的土壤理化性质均下降。土壤固氮酶活以撂荒土壤最高,G2轮作最低。土壤固氮微生物nifH基因多样性以G1轮作最高、G2轮作最低,门水平上以变形菌门(Proteobacteria)是优势共有nifH基因类群,相对丰度占群落的63.0%-92.4%;属水平上,偶氮氢单胞菌属(Azohydromonas)是不同轮作制度下的优势物种,慢生根瘤属(Brad...     

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.

     

美国白蛾在我国发生的世代变化分析

美国白蛾是原产于北美地区的世界性植物检疫性害虫,自1979年发现入侵我国以来,严重威胁我国的生态安全。近年来,随着全球气候变暖,美国白蛾疫情不断南扩,其化性在一些地区出现新的变化。本文根据1979—2021年该虫发生扩散情况和各地区监测普查结果,结合部分地区温度变化数据,全面分析了美国白蛾入侵我国以来的发生扩散过程,对比分析了各地区发生世代数及变化情况,阐明部分地区世代数增加的原因。针对美国白蛾世代数演变规律和反复暴发成灾的情况,提出了当前应关注的几个重点问题:世代数增加引发的扩张速率显著上升;扩张潜力与沿长江地区扩散趋势减缓关系;暴发成灾时有发生与本土化趋势不明显。鉴于当前外来入侵物种管理要求提高及多年来美国白蛾防控工作中暴露的一些问题,提出要在守护国家生物安全底线前提下制定防治策略、在统筹生物多样性保护下实现控制目标、在做好应急准备下摸清暴发成灾规律、在强化疫情检疫管理中提高疫情认定效率等对策。     

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.     

九种蹄盖蕨科植物配子体的酯酶同工酶分析

本文培养了蹄盖蕨科９个种即中华蹄盖蕨（Ａｔｈｙｒｉｕｍｓｉｎｅｎｓｅ）带岭蹄盖蕨（Ａ．ｄａｌｉｎｇｅｎｓｅ）、多齿蹄盖蕨（Ａ．ｍｕｌｔｉｄｅｎｔａｔｕｍ）（包括青柄和紫柄２种带岭蹄蕨类型）、东北蛾眉蕨（Ｌｕｎａｔｈｙｒｉｕｍｐｙｃｎｏｓｏｒｕｍ）、朝鲜介蕨（Ｄｒｙｏａｔｈｙｒｉｕｍｃｏｒｅａｎｕｍ）、山冷蕨（Ｃｙｓｔｏｐｔｅｒｉｓｓｕｄｅｔｉｃａ）、假冷蕨（Ｐｓｅｕｄｏｃｙｓｔｏｐｔｅｒｉｓｓｐｉｎｕｌｏｓａ）、欧洲羽节蕨（Ｇｙｍｎｏｃａｒｐｉｕｍｄｒｙｏｐｔｅｒｉｓ）和黑鳞短肠蕨（Ａｌａｎｔｏｄｉａｃｒｅｎａｔａ）的配子体为材料,进行酯酶同工酶的分析,以表明它们的种间差异,其中多齿蹄盖蕨的青柄和紫柄两种类型的酶谱存在明显差异,应考虑紫柄为多齿蹄盖蕨的变型。     

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。