体外重构STUB1介导α-1抗胰蛋白酶突变体Z蛋白泛素化修饰反应体系

α-1抗胰蛋白酶Z型突变体蛋白(α-1 antitrypsin Z-mutant protein, ATZ)是引发α-1抗胰蛋白酶缺陷症(α-1 antitrypsin deficiency, AATD)的主要原因,研究ATZ蛋白的泛素化修饰和降解对于治疗AATD具有重要意义。STUB1是一种重要的E3泛素连接酶,参与调节多种蛋白质的泛素化修饰。然而,STUB1是否参与ATZ的泛素化修饰尚未明确。本研究首先将ATZ和STUB1的编码基因克隆到pET28a质粒,构建了这2个蛋白的表达质粒。随后,将重组质粒转入大肠杆菌表达系统,在优化诱导条件实现了重组蛋白的异源表达。通过金属螯合亲和层析技术纯化得到目的蛋白,并通过蛋白质谱分析验证了其氨基酸序列的准确性。利用纯化的ATZ和STUB1重组蛋白,构建了一个体外泛素化修饰反应体系。实验结果显示,在ATP、E1泛素激活酶和E2泛素结合酶的协同作用下,STUB1成功催化了ATZ的泛素化修饰。本研究提供了一种体外获得Z型突变体ATZ纯化蛋白的方法,并确认了STUB1介导ATZ的泛素化修饰功能,推进了对α-1抗胰蛋白酶Z型突变体蛋白在细胞内降解过程的调控机制的理解。     

不同老化程度的不结球白菜种子活力指标变化及其相关分析

以不结球白菜品种‘高梗白’种子为材料,采用高温(42℃)高湿(相对湿度100%)人工加速老化处理,研究不同老化程度下种子活力指标的变化及其相关性。结果显示:(1)发芽指标(除不正常苗率)和出苗指标均随种子老化程度的加深而显著下降,不正常苗率显著上升。(2)随老化程度的加深,种子中超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性显著下降;种子的超氧阴离子(O2.-)产生速率先增高后降低,过氧化氢(H2O2)含量逐渐增加,丙二醛(MDA)含量下降,种子浸出液相对电导率升高;种子的可溶性糖含量、可溶性蛋白含量和脱氢酶活性下降,α-淀粉酶活性先升高后降低。(3)老化种子的发芽势、发芽率、SOD活性、H2O2含量、相对电导率和可溶性糖含量与出苗指标间均存在极显著相关性。研究表明,不结球白菜种子的发芽势、发芽率、SOD活性、H2O2含量和可溶性糖含量随老化程度加深的变化规律一致,且指标间以及与出苗指标均有极显著相关性,可以作为检验不结球白菜种子活力的候选指标。     

云南省农田生态系统碳足迹时空变化及其影响因素

农田碳足迹研究对农田生态系统管理与农业可持续发展具有重要意义,也可表征农田扩展的生态影响程度。利用县域尺度统计数据与空间分析,对云南省农田生态系统近30年的碳足迹的时空演变进行研究。结果表明:1985—2015年期间,云南省农田生态系统碳排放年均增幅为13.9%,化肥施用引起的碳排放贡献率最大,为56%,2015年的化肥单位面积碳排放达到331.6kg/hm2。云南省农田生态系统碳吸收年均增幅为3.04%,稻谷的碳吸收比例最大,为41%,然而,玉米的碳吸收的增幅最大,为8.76%。云南省农田生态系统存在碳生态盈余,且碳足迹总体呈现增长趋势,年均增长率为16.8%,单位面积碳足迹随年份增加不断增长。从空间上看,云南省农田生态系统碳排放、碳吸收在空间上均呈东南高、西北低的分布格局,而碳足迹在空间上呈现东西部高、中部低的分布格局,三者的空间差异和变化幅度差异都较大。     

野生大豆种子蛋白含量差异的生理及结构基础的探讨

利用ＳＤＳ－聚丙烯酰胺凝胶电泳、电子显微镜、蛋白及酰脲含量测定等技术,对高蛋白含量（５０．７％ ）的５０３５９ 和低蛋白含量（４０．８％ ）的５０３０５ 两个野生大豆在种子发育过程中贮藏蛋白积累的速率、蛋白组分合成的起始时间、蛋白体发育的进程以及幼茎的酰脲含量进行了比较研究。结果表明：野生大豆５０３５９的高蛋白含量是与其种子发育过程中较高的植株酰脲含量、较早较快的贮藏蛋白合成及积累速率,液泡中高效的蛋白贮藏方式以及蛋白体在子叶细胞中占有较大体积相联系的     

Typha as a wetland food resource: evidence from the Tianluoshan site,Lower Yangtze Region,China

Vegetation History and Archaeobotany - Wetlands have been attractive environments for early communities worldwide. In China, wetlands offered natural ecological settings for the start of rice...     

Transformation of five grape rootstocks with plant virus genes and a virE2 gene from Agrobacterium tumefaciens

Summary To facilitate the development of transgenic grapevines that are resistant to grapevine fanleaf virus (GFLV), grapevine leafroll-associated closterovirus (GLRaV-3) and crown gall diseases, we developed a rapid system for regenerating root-stocks: Couderc 3309, Vitis riparia ‘Gloire de Montpellier’, Teleki 5C, Millardet et De Grasset 101-14, and 110 Richter via somatic embryogenesis. Embryo culture and grape regeneration were accomplished with four media. Embryogenic calluses from anthers were induced in the initiation medium [MS basic medium containing 20 g sucrose per L, 1.1 mg 2,4-dichlorophenoxyacetic acid (2,4-D) per L, 0.2 mg N⁶-benzyladenine (BA) per L, and 0.8% Noble agar). The percentage of anthers that developed into embryogenic calli ranged from 2 to 16.3% depending on the rootstock. Calluses with early globular stage embryos were cocultivated with Agrobacterium tumefaciens strain C58Z707 containing the gene constructs of interest. The genes were sense-oriented translatable and antisense coat protein genes from GFLV and GLRaV-3, a truncated HSP90-related gene of GLRaV-3 (43K), and a virE2 del B gene from A. tumefaciens strain C58. Twenty independent transformation experiments were performed on five rootstocks. After 3–4 mo. under kanamycin selection, secondary embryos were recovered on differentiation medium (1/2 MS salts with 10 g sucrose per L, 4.6 g glycerol per L, and 0.8% Noble agar). Embryos that were transformed were regenerated on a medium containing MS salts with 20 g sucrose per L, 4.6 g glycerol per L, 1 g casein hydrolysate per L, and 0.8% Noble agar. Elongated embryos were then transferred to a rooting medium supplemented with 0.1 mg BA per L, 3 g activated charcoal per L, 1.5% sucrose, and 0.65% Bacto agar. A total of 928 independent putative transgenic plants were propagated in the greenhouse. All plants were tested for neomycin phosphotransferase II expression by enzyme-linked immunosorbent assay (ELISA). The presence of transgenes was assessed by polymerase chain reaction and Southern analysis. ELISA revealed various levels of expression of GFLV coat protein in transgenic plants of Couderc 3309. The transgenic rootstocks that have been generated are being screened to determine whether transgenes have conferred resistance to the virus and crown gall diseases.     

鄂尔多斯高原西部几种荒漠灌丛群落种间联结关系的研究

采用ＰＣ、ＰＣＣ、ＡＣ、Ｘ２和ｒ等公式对鄂尔多斯高原西部５种典型荒漠灌丛群落种间联结系数的求算,通过这些种间的相互关系,阐述了这些物种对生境要求的差异和分布特点,本文的研究结果对与建立在种群关系基础上的荒漠植被的研究具有重要的意义。     

两个污水处理系统的能值与经济综合分析

运用改进的能值评价指标和经济指标对构建的两个污水处理综合系统("污水处理厂处理+脱水污泥填埋"及"污水处理厂处理+脱水污泥填埋+中水回用")进行了环境可持续性和经济竞争力的综合分析。改进的能值指标(能值产出率SEYR、环境负载率SELR、环境可持续发展指数SESI和能值受益率SEBR)是在考虑研究对象废物及产物能值对环境影响的基础上提出的,更好地反映了系统的特征。结果表明,中水回用系统的增加有利于提升系统的环境可持续发展能力;"污水处理厂处理+脱水污泥填埋+中水回用"系统较"污水厂处理+脱水污泥填埋"系统的经济竞争力差,但却拥有更强的可持续发展能力,有利的经济政策能够刺激其更好的推广运用。     

Genome‐wide detection of CNVs associated with beak deformity in chickens using high‐density 600K SNP arrays

Beak deformity (crossed beaks) is found in several indigenous chicken breeds including Beijing‐You studied here. Birds with deformed beaks have reduced feed intake and poor production performance. Recently, copy number variation (CNV) has been examined in many species and is recognized as a source of genetic variation, especially for disease phenotypes. In this study, to unravel the genetic mechanisms underlying beak deformity, we performed genome‐wide CNV detection using Affymetrix chicken high‐density 600K data on 48 deformed‐beak and 48 normal birds using penncnv . As a result, two and eight CNV regions (CNVRs) covering 0.32 and 2.45 Mb respectively on autosomes were identified in deformed‐beak and normal birds respectively. Further RT‐qPCR studies validated nine of the 10 CNVRs. The ratios of six CNVRs were significantly different between deformed‐beak and normal birds (P < 0.01). Within these six regions, three and 21 known genes were identified in deformed‐beak and normal birds respectively. Bioinformatics analysis showed that these genes were enriched in six GO terms and one KEGG pathway. Five candidate genes in the CNVRs were further validated using RT‐qPCR. The expression of LRIG2 (leucine rich repeats and immunoglobulin like domains 2) was lower in birds with deformed beaks (P < 0.01). Therefore, the LRIG2 gene could be considered a key factor in view of its known functions and its potential roles in beak deformity. Overall, our results will be helpful for future investigations of the genomic structural variations underlying beak deformity in chickens.