The N-terminal region of the starch-branching enzyme from Phaseolus vulgaris L. is essential for optimal catalysis and structural stability

Starch-branching enzymes (SBEs) play a pivotal role in determining the fine structure of starch by catalyzing the syntheses of alpha-1,6-branch points. They are the members of the alpha-amylase family and have four conserved regions in a central (beta/alpha)8 barrel, including the catalytic sites. Although the role of the catalytic barrel domain of an SBE is known, that of its N- and C-terminal regions remain unclear. We have previously shown that the C-terminal regions of the two SBE isozymes (designated as PvSBE1 and PvSBE2) from kidney bean (Phaseolus vulgaris L.) have different roles in branching enzyme activity. To understand the contribution of the N-terminal region to catalysis, six chimeric enzymes were constructed between PvSBE1 and PvSBE2. Only one enzyme (1Na/2Nb)-II, in which a portion of the N-terminal region of PvSBE2 was substituted by the corresponding region of PvSBE1, retained 6% of the PvSBE2 activity. The N-terminal truncated form (DeltaN46-PvSBE2), lacking 46 N-terminal residues of PvSBE2, lost enzyme activity and stability to proteolysis. To investigate the possible function of this region, three residues (Asp-15, His-24, and Arg-28) among these 46 residues were subjected to site-directed mutagenesis. The purified mutant enzymes showed nearly the same K(m) values as PvSBE2 but had lower V(max) values and heat stabilities than PvSBE2. These results suggest that the N-terminal region of the kidney bean SBE is essential for maximum enzyme activity and thermostability.     

A secondary xylan-binding site enhances the catalytic activity of a single-domain family 11 glycoside hydrolase

Bacillus circulans xylanase (BcX) is a single-domain family 11 glycoside hydrolase. Using NMR-monitored titrations, we discovered that an inactive variant of this enzyme, E78Q-BcX, bound xylooligosaccharides not only within its pronounced active site (AS) cleft, but also at a distal surface region. Chemical shift perturbation mapping and affinity electrophoresis, combined with mutational studies, identified the xylan-specific secondary binding site (SBS) as a shallow groove lined by Asn, Ser, and Thr residues and with a Trp at one end. The AS and SBS bound short xylooligosaccharides with similar dissociation constants in the millimolar range. However, the on and off-rates to the SBS were at least tenfold faster than those of kon approximately 3x10(5) M(-1) s(-1) and koff approximately 1000 s(-1) measured for xylotetraose to the AS of E78Q-BcX. Consistent with their structural differences, this suggests that a conformational change in the enzyme and/or the substrate is required for association to and dissociation from the deep AS, but not the shallow SBS. In contrast to the independent binding of small xylooligosaccharides, high-affinity binding of soluble and insoluble xylan, as well as xylododecaose, occurred cooperatively to the two sites. This was evidenced by an approximately 100-fold increase in relative Kd values for these ligands upon mutation of the SBS. The SBS also enhances the activity of BcX towards soluble and insoluble xylan through a significant reduction in the Michaelis KM values for these polymeric substrates. This study provides an unexpected example of how a single domain family 11 xylanase overcomes the lack of a carbohydrate-binding module through the use of a secondary binding site to enhance substrate specificity and affinity.     

Harvest prediction in ‘Algerie’ loquat

Plant phenology is in great measure driven by air temperature. To forecast harvest time for ‘Algerie’ loquat accurately, the growing degree days (GDD) needed from bloom to ripening were determined using data from nine seasons. The methods proposed by Zalom et al. (Zalom FG, Goodell PB, Wilson LT, Barnett WW, Bentley W, Degree-days: the calculation and use of heat units in pest management, leaflet no 21373, Division Agriculture and Natural Resources, University of California 10 pp, 1983) were compared as regards their ability to estimate heat summation based on hourly records. All the methods gave remarkably similar results for our cultivation area, although the double-sine method showed higher performance when temperatures were low. A base temperature of 3°C is proposed for ‘Algerie’ loquat because it provides a coefficient of variation in GDD among seasons of below 5%, and because of its compatibility with loquat growth. Based on these determinations, ‘Algerie’ loquat requires 1,715 GDD from bloom to harvest; under our conditions this heat is accumulated over an average of 159 days. Our procedure permits the ‘Algerie’ harvest date to be estimated with a mean error of 4.4 days (<3% for the bloom-harvest period). GDD summation did not prove superior to the use of the number of calendar days for predicting ‘Algerie’ harvest under non-limiting growing conditions. However, GDD reflects the developmental rate in water-stressed trees better than calendar days. Trees under deficit irrigation during flower development required more time and more heat to ripen their fruits.     

次生林概念与生态干扰度

近一个世纪强烈的人为干扰使世界范围内的原始林面积锐减,次生林已成为中国乃至世界森林资源的主体.虽然在文献与现实中"次生林（Secondary forest）"被广泛使用,但次生林的概念在各个国家,甚至同一国家或地区以及各种不同文献中存在相互矛盾的情况,这给次生林研究与经营实践带来诸多不便;而不准确的次生林定义也为与各个层次的决策制定者及公众之间的交流带来了障碍.本文在查阅大量国内外关于次生林概念文献的基础上,结合近年来关于次生林生态与经营研究实践,综合分析了次生林概念的不确定性,同意以往次生林定义中的基本内涵,并认为：次生林是由于人为破坏性干扰或异常自然干扰使原始林固有的林分结构、物种组成或基本功能发生了显著变化,经过天然更新或人工诱导天然更新恢复形成的林分.但该定义中仍存在着很多不确定性, 如:怎样的干扰为"人为破坏性干扰"和"异常自然干扰";"林分结构、物种组成或基本功能"发生了怎样的变化为"显著变化";"人工诱导天然更新"中"人工"参与的成分比例如何等.次生林概念是在20世纪初由植物演替学家提出,当时未考虑干扰的持续性;而实际上,无论是次生林还是原始林,均是森林生态系统演替过程中的某一状态,在现代森林生态系统研究中,应重新规范"次生林"的概念.考虑到"次生林"定义的不确定性或困难性,建议使用"森林自然度"或"森林生态系统成熟度"或"森林干扰度"来表达现实森林所处的状态,但该方面的研究与实践尚需加强.     

一类无穷时滞微分系统的周期解和全局吸引性

考虑如下具有无穷时滞的微分系统:x′(t)=A(t,x(t))x(t) (integral from -∞to 0)f(t,s,x(s t))ds (sum from i=1 to p) fi(t,x(t-T_i(t)))的周期解.利用重合度理论和构造适当的Lyapunov泛函得到上述系统周期解存在性和全局吸引性的充分条件,推广了已有的结论,得到了新的结果.     

秦岭南坡锐齿栎次生林空间结构特征分析

该研究采用坐标定位方法对秦岭南坡不同坡向锐齿栎次生林样地内胸径DBH≥5cm的乔木进行调查,记录树种名称、坐标、胸径、高度、冠幅、生长状况等因子,运用角尺度、大小比数和混交度3个指数分析不同坡向锐齿栎次生林的空间结构,探寻不同坡向锐齿栎林空间结构的形成和差异。结果显示:(1)调查样地内物种多样性丰富,锐齿栎重要值最大,长势良好,占据主要生态空间。(2)两种坡向中,角尺度取值为0.5的个体数均过半,表明随机分布是锐齿栎次生林的主要分布方式。(3)优势木、亚优势木、中庸木、劣态木和绝对劣态木所占比重均为20%左右,表明林木大小差异不明显,在空间结构单元中,林木较稳定,阳坡优势木个体数高于阴坡。(4)两种坡向中,混交度取值大于0.5林木个体分别为84%和73%,表明多数林木个体处于中度、强度和极强度混交状态。研究表明,秦岭南坡锐齿栎次生林聚集度不高、混交良好、分化程度较高,锐齿栎种群优势度明显,林分整体暂时处于相对稳定状态。     

全程无缝隙护理模式在护理质量管理中的应用

目的 探讨全程无缝隙护理模式对肿瘤科护理工作质量的影响。方法 2013年1月-12月我院肿瘤科采用无缝隙护理模式的患者120例为观察组,2012年1月-12月采用常规护理的120例肿瘤科患者为对照组,对比两组患者的心理舒适度、患者对护理工作的满意度。结果 观察组患者的安全感、满足感、尊重感所占比例均高于对照组,差异均有统计学意义（P＜0.05）。观察组患者对护士的工作能力、关爱与沟通、服务热情主动、病区管理、健康教育满意度均高于对照组,差异均有统计学意义（P＜0.05）。结论 全程无缝隙护理模式应用于肿瘤科护理,能够有效改善患者不良情绪,提高护理工作质量及患者满意度。     

Expression of a bacterial 3‐dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency

Lignin confers recalcitrance to plant biomass used as feedstocks in agro‐processing industries or as source of renewable sugars for the production of bioproducts. The metabolic steps for the synthesis of lignin building blocks belong to the shikimate and phenylpropanoid pathways. Genetic engineering efforts to reduce lignin content typically employ gene knockout or gene silencing techniques to constitutively repress one of these metabolic pathways. Recently, new strategies have emerged offering better spatiotemporal control of lignin deposition, including the expression of enzymes that interfere with the normal process for cell wall lignification. In this study, we report that expression of a 3‐dehydroshikimate dehydratase (QsuB from Corynebacterium glutamicum) reduces lignin deposition in Arabidopsis cell walls. QsuB was targeted to the plastids to convert 3‐dehydroshikimate – an intermediate of the shikimate pathway – into protocatechuate. Compared to wild‐type plants, lines expressing QsuB contain higher amounts of protocatechuate, p‐coumarate, p‐coumaraldehyde and p‐coumaryl alcohol, and lower amounts of coniferaldehyde, coniferyl alcohol, sinapaldehyde and sinapyl alcohol. 2D‐NMR spectroscopy and pyrolysis‐gas chromatography/mass spectrometry (pyro‐GC/MS) reveal an increase of p‐hydroxyphenyl units and a reduction of guaiacyl units in the lignin of QsuB lines. Size‐exclusion chromatography indicates a lower degree of lignin polymerization in the transgenic lines. Therefore, our data show that the expression of QsuB primarily affects the lignin biosynthetic pathway. Finally, biomass from these lines exhibits more than a twofold improvement in saccharification efficiency. We conclude that the expression of QsuB in plants, in combination with specific promoters, is a promising gain‐of‐function strategy for spatiotemporal reduction of lignin in plant biomass.     

N-酰化低聚壳聚糖衍生物还原能力的研究

对低聚壳聚糖进行N-酰化改性,制得取代度相同的N-马来酰低聚壳聚糖(NMCOS),N-琥珀酰低聚壳聚糖(NSCOS),N-邻苯二甲酰低聚壳聚糖(NPCOS),其中NMCOS1、NSCOS1、NPCOS1的取代度均为0.25;NMCOS2、NSCOS2、NPCOS2的取代度均为0.49。考察了6种N-酰化低聚壳聚糖衍生物的还原能力。结果表明:当取代度相同时,N-邻苯二甲酰低聚壳聚糖的还原能力最强,其次是N-马来酰低聚壳聚糖,N-琥珀酰低聚壳聚糖的还原能力最差。这可能是由取代基的性质不同所致。     

苏南地区典型城镇建设用地扩展的时空分异

选择江苏省常熟市南部的辛庄镇为研究区,采用该区1980、1991、2001和2009年4期高分辨率遥感影像,利用GIS技术,结合现有的建设用地扩展指标构建了综合扩展程度指数模型,定量分析了研究区1980-2009年间3个时段建设用地扩展的总体特征和时空分异特征.结果表明:随着农村城镇化和工业化进程的加快,辛庄镇建设用地显著扩展,1980-2009年间共增加19.24 km2,其中,2001-2009年该区建设用地进入高速扩展期,扩展面积、扩展贡献率和扩展强度均最大.研究区建设用地扩展具有明显的空间分异特征:1980-1991年,新增建设用地主要集中于镇区所在村域,1991年后,辛庄镇建设重心逐渐向工业发达的村庄转移.研究期间,辛庄镇的新增建设用地主要源于水田和旱地,二者共占建设用地总增加面积的88.1%,其余土地类型对建设用地的转入贡献率相对较小.