龙眼内源激素变化和花芽分化及大小年结果的关系

本文探讨了隔年结果的龙眼花芽和营养芽内源激素在成花过程中的作用及其与结果大小年的关系．结果表明：大年树的细胞分裂素ｉＰＡ明显地高于小年树,而ＧＡ和ＡＢＡ含量明显低于小年树,说明细胞分裂素有利于龙眼花芽分化,ＧＡ和ＡＢＡ不利于花芽分化．龙眼大年树细胞分裂素和赤霉素的比值显著高于小年树．外施ＰＰ３３３能促进花芽分化,具有缩短花序、提高着果率和增加产量的作用．     

The process of embryo abortion of stenospermocarpic grape and it develops into plantlet in vitro using embryo rescue

Plant Cell, Tissue and Organ Culture (PCTOC) - The fruit of ‘Dangshansuli’ pear is yellowish green in colour, while that of its mutant ‘Xiusu’ is russet in colour. A...     

Fast protein sequencing of monoclonal antibody by real-time digestion on emitter during nanoelectrospray

Growth in the pharmaceutical industry has led to an increasing demand for rapid characterization of therapeutic monoclonal antibodies. The current methods for antibody sequence confirmation (e.g., N-terminal Edman sequencing and traditional peptide mapping methods) are not sufficient; thus, we developed a fast method for sequencing recombinant monoclonal antibodies using a novel digestion-on-emitter technology. Using this method, a monoclonal antibody can be denatured, reduced, digested, and sequenced in less than an hour. High throughput and satisfactory protein sequence coverage were achieved by using a non-specific protease from Aspergillus saitoi, protease XIII, to digest the denatured and reduced monoclonal antibody on an electrospray emitter, while electrospray high voltage was applied to the digestion mixture through the emitter. Tandem mass spectrometry data was acquired over the course of enzyme digestion, generating similar information compared to standard peptide mapping experiments in much less time. We demonstrated that this fast protein sequencing method provided sufficient sequence information for bovine serum albumin and two commercially available monoclonal antibodies, mouse IgG1 MOPC21 and humanized IgG1 NISTmAb. For two monoclonal antibodies, we obtained sequence coverage of 90.5–95.1% for the heavy chains and 98.6–99.1% for the light chains. We found that on-emitter digestion by protease XIII generated peptides of various lengths during the digestion process, which was critical for achieving sufficient sequence coverage. Moreover, we discovered that the enzyme-to-substrate ratio was an important parameter that affects protein sequence coverage. Due to its highly automatable and efficient design, our method offers a major advantage over N-terminal Edman sequencing and traditional peptide mapping methods in the identification of protein sequence, and is capable of meeting an ever-increasing demand for monoclonal antibody sequence confirmation in the biopharmaceutical industry.     

Phosphoproteomic analysis reveals site-specific changes in GFAP and NDRG2 phosphorylation in frontotemporal lobar degeneration

Frontotemporal lobar degeneration (FTLD) is a progressive neurodegenerative disease characterized by behavioral abnormalities, personality changes, language dysfunction, and can co-occur with the development of motor neuron disease. One major pathological form of FTLD is characterized by intracellular deposition of ubiquitinated and phosphorylated TAR DNA binding protein-43 (TDP-43), suggesting that dysregulation in phosphorylation events may contribute to disease progression. However, to date systematic analysis of the phosphoproteome in FTLD brains has not been reported. In this study, we employed immobilized metal affinity chromatography (IMAC) followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify phosphopeptides from FTLD and age-matched control post-mortem human brain tissue. Using this approach, we identified 786 phosphopeptides in frontal cortex (control and FTLD), in which the population of phosphopeptides represented approximately 50% of the total peptides analyzed. Label-free quantification using spectral counts revealed six proteins with significant changes in the FTLD phosphoproteome. N-myc-Downstream regulated gene 2 (NDRG2) and glial fibrillary acidic protein (GFAP) had an increased number of phosphospectra in FTLD, whereas microtubule associated protein 1A (MAP1A), reticulon 4 (RTN4; also referred to as neurite outgrowth inhibitor (Nogo)), protein kinase C gamma (PRKCG), and heat shock protein 90 kDa alpha, class A member 1(HSP90AA1) had significantly fewer phosphospectra compared to control brain. To validate these differences, we examined NDRG2 phosphorylation in FTLD brain by immunoblot analyses, and using a phosphoserine-13 (pSer13) GFAP monoclonal antibody we show an increase in pSer13 GFAP levels by immunoblot concomitant with increased overall GFAP levels in FTLD cases. These data highlight the utility of combining proteomic and phosphoproteomic strategies to characterize post-mortem human brain tissue.     

蜀南竹海毛竹林土壤物理性质空间异质性

土壤空间异质性是土壤的重要综合属性,对土壤的结构与功能以及植被的空间格局有重要影响。以蜀南竹海核心区毛竹林土壤为研究对象,采用1∶10000地形图为工作底图,按公里网格布点,应用地统计学方法,分析了土壤物理性质的空间异质性,旨在为研究区毛竹林的可持续经营提供基础数据和理论参考。结果表明:除土壤最大持水量和最小持水量外,研究区土壤粘粒、容重、非毛管孔隙度、毛管孔隙度和毛管持水量等5个土壤物理性质空间变异性明显,其半变异函数用指数、高斯和球状模型拟合,变程范围在1252.3—11887.1 m,块金值和基台值比值范围为0.03%—52.07%,表明各物理性质有不同程度的空间自相关性;土壤持水特性有较大的块金值,说明存在小尺度的空间异质性;普通克里格插值及物理性质空间分布图表明,土壤物理性质在空间分布上具有斑块性,在不同土层深度的空间分布有一致性。人为干扰是影响研究区土壤物理性质空间分布格局的一个主要原因。减少人为干扰,优化林分结构,维护与提高毛竹林生产力。     

无核香味葡萄胚挽救育种研究

为创制无核香味葡萄新种质,该研究以7个无核葡萄品种为母本,4个玫瑰香味品种为父本,配置了9个杂交组合。田间杂交后,通过胚挽救技术离体培养杂种胚珠,使胚珠中幼胚继续发育,继而在幼胚萌发成苗后利用无核标记对F1代株系进行无核性状的早期检测,同时对杂种幼苗进行温室炼苗,最终移栽大田。结果表明:(1)利用胚挽救技术获得365个胚挽救株系,移栽大田并成活182株杂种苗。(2)对其中72个株系利用无核基因探针GSLP1-569与无核分子标记SCF27-2000进行无核性状检测,分别检测出8个和38个杂种后代携带无核特异性条带,初步确定为无核株系。(3)杂交组合中以‘红宝石无核’、‘火焰无核’、‘昆香无核’为母本的组合成苗率相对较高,较适宜做母本;香味品种中‘玫瑰香’更适合做为父本选育香味无核种质。(4)以‘火焰无核’和‘昆香无核’为母本的杂交组合分别在授粉后的43d和51d采样,胚发育率(6.96%~8.54%和16.92%~18.13%)和成苗率(6.45%和8.75%~10%)较高;‘红宝石无核’ב玫瑰香’、‘莫莉莎无核’ב玫瑰香’和‘波尔莱特’ב玫瑰香’3个组合分别在授粉后59~60d、54d、52d采样,胚挽救育种效果较好;杂交组合‘克瑞森无核’ב巨玫瑰’和‘奇妙无核’ב昆香无核’胚败育时期分别为授粉后39d和48d。(5)幼胚发育至子叶型胚时最易成苗,延长胚珠离体暗培养时间显著增加了畸形胚比率。研究认为,葡萄胚挽救育种过程中,胚珠离体培养时间在8~10周为宜。     

Quantitative proteomics of intracellular Porphyromonas gingivalis

Whole-cell quantitative proteomic analyses were conducted to investigate the change from an extracellular to intracellular lifestyle for Porphyromonas gingivalis, a Gram-negative intracellular pathogen associated with periodontal disease. Global protein abundance data for P. gingivalis strain ATCC 33277 internalized for 18 h within human gingival epithelial cells and controls exposed to gingival cell culture medium were obtained at sufficient coverage to provide strong evidence that these changes are profound. A total of 385 proteins were overexpressed in internalized P. gingivalis relative to controls; 240 proteins were shown to be underexpressed. This represented in total about 28% of the protein encoding ORFs annotated for this organism, and slightly less than half of the proteins that were observed experimentally. Production of several proteases, including the classical virulence factors RgpA, RgpB, and Kgp, was decreased. A separate validation study was carried out in which a 16-fold dilution of the P. gingivalis proteome was compared to the undiluted sample in order to assess the quantitative false negative rate (all ratios truly alternative). Truly null (no change) abundance ratios from technical replicates were used to assess the rate of quantitative false positives over the entire proteome. A global comparison between the direction of abundance change observed and previously published bioinformatic gene pair predictions for P. gingivalis will assist with future studies of P. gingivalis gene regulation and operon prediction.     

‘火州黑玉’葡萄杂交后代果实性状遗传倾向分析

为了探索‘火州黑玉’葡萄果实性状遗传规律,2013~2014年,对‘火州黑玉’为母本的3个组合198个杂交后代的果实性状(包括核性、果色、香味、质地、可溶性固形物含量、果粒重和果粒形状)进行了分析研究。结果表明:后代中胚败育型株系占77.3%,有香味株系仅1个;杂交后代的果色、质地、可溶性固形物含量、果粒重、果形指数呈现数量遗传性状且连续广泛分离;果粒表现无核、深果色、脆肉、小粒、圆形等性状具有遗传倾向;可溶性固形物含量遗传较复杂。研究认为,‘火州黑玉’葡萄能将无核、深果色、脆质地、圆果形等性状以很强优势传递给F1代,但很难出现有香味株系;果粒呈变小趋势,但存在有选择大果型株系的潜力。