红壤溶磷菌的筛选及溶磷机制

采用以磷酸铝为磷源的蒙金娜(PVK)液体培养基研究了从红壤土中筛选出的4种溶磷菌的溶磷效果,选出其中的优势菌株B1,并对其溶磷机理做出初步探讨。结果表明,所筛选出的4株溶磷菌在液体培养条件下均有显著的溶磷效果,其中菌株B1在培养4 d后有效溶磷量最大,达到292.8 mg L-1。各处理培养液pH在培养期间均有显著下降,pH从7.0下降至3.2~4.7。高效液相色谱测定发现,各菌株培养液中有机酸的种类与含量随培养时间变化而不同,其中菌株B1主要分泌草酸和苹果酸,培养1 d后有机酸总量可达到5 mmol L-1;通过添加有机酸对磷酸铝活化的试验表明,分泌有机酸溶磷仅是菌株B1溶磷机制之一,可能还存在其他溶磷机制。菌株B1生长的适宜pH范围为5~9,最适培养温度为30℃,100 ml三角瓶的最适装液量为30~40 ml。经鉴定,菌株B1与苏云金芽孢杆菌有99.9%的相似性。     

蜡样芽孢杆菌及其生物被膜对有机酸及乙醇的抗性比较

为比较蜡样芽孢杆菌及其生物被膜对环境压力的抗性,该文主要研究了4种有机酸（乙酸、柠檬酸、乳酸和苹果酸）和乙醇对蜡样芽孢杆菌及其生物被膜存活率的影响。结果表明：生物被膜态蜡样芽孢杆菌对乙酸的抗性高于浮游态菌。扫描电镜结果显示,经乙酸处理后,浮游态蜡样芽孢杆菌的细胞表面严重受损,而生物被膜态菌的表面形态未发生明显变化。在柠檬酸、乳酸、苹果酸和乙醇存在的条件下,生物被膜态蜡样芽孢杆菌比浮游态菌表现出更强的压力抗性,特别是在高浓度（有机酸16%～20%,乙醇50%～60%）的情况下,该现象尤为显著。因此,在食品工业中控制被膜态蜡样芽孢杆菌对预防和阻止食品腐败非常重要。该研究结果为实际生产中有效控制蜡样芽孢杆菌及其生物被膜的形成提供参考。     

Environmental effects on soil NO concentrations and root N uptake in beech and spruce forests

This study aimed to investigate the shifts in net nitrogen (N) uptake and N compounds of fine roots over the vegetation period (i.e., spring, summer, autumn) and correlate this with NO concentration in the soil. Soil NO concentration was measured using gas lysimeters for collection and a chemiluminescence analyzer for quantification. Net N uptake by the roots was determined using the ¹⁵N enrichment technique. N pools were quantified using spectrophotometric techniques. Soil NO concentrations at beech and spruce forest sites were highest in spring (June), and lowest in winter (December). Total N of the roots was similar during the seasons and between the two years under study despite considerable variation of different N compounds. Net N uptake generally increased with higher N supply. Correlation analysis revealed a positive relationship between soil NO concentration and net N uptake only for spruce trees. This relationship seemed to be modulated by environmental factors and tree species.     

基于氨基酸组成的黄酒酒龄电子舌鉴别

该研究采用电子舌结合化学计量学方法用于黄酒酒龄的快速鉴别。为确证黄酒样品酒龄,采用氨基酸分析仪分析了1年陈、3年陈和5年陈黄酒中20种氨基酸,并利用主成分分析（principal component analysis,PCA）对氨基酸数据进行了分析。采用电位型电子舌采集了不同酒龄黄酒样品的味觉指纹信息,并采用判别分析（discriminant analysis,DA）方法结合味觉指纹信息建立黄酒酒龄快速鉴别模型。采用偏最小二乘法（partial least squares regression,PLSR）建立电子舌响应信号与氨基酸含量之间的相关关系。氨基酸数据结合PCA分析表明所有样品均标注正确;电子舌结合DA所建黄酒酒龄鉴别模型可将3个年份预测集样品正确区分;异亮氨酸（Ile）、天门冬氨酸（Asp）、酪氨酸（Tyr）和缬氨酸（Val）与电子舌相关性高,模型的相对分析误差（Residual predictive deviation, RPD）高于2。研究表明电位型电子舌结合判别分析是黄酒龄鉴别的稳健方法。     

普鲁兰酶氮端氨基酸的截短对其酶学性质的影响

淀粉是丰富的生物质资源,已被广泛地应用到食品、酿酒、医药等各个相关的领域.普鲁兰酶(pullulanase,pulA,EC 3.2.1.41)作为淀粉脱支酶,可以水解淀粉中的α-1,6糖苷键,最大限度地降解淀粉原料,提高淀粉利用率.因此,寻找一条国产化的道路开发我国自己的普鲁兰酶,具有长远的意义.课题组前期从淀粉厂附近的土壤中筛到一株普鲁兰酶的高产菌株变栖克雷伯氏菌(Klebiella variicola)strain 7,克隆获得pulA基因(GenBank登录号:KJ146839.1)后在大肠杆菌(Escherichia coli) BL21 (DE3)中异源表达.为提高该蛋白的表达量和分泌效率,同时改善普鲁兰酶的性质,本研究利用基因工程的手段构建了普鲁兰酶去信号肽突变体M1和氮端31个氨基酸的截短突变体M2,结果显示M1和M2最适温度均为45 ℃,二者的最适pH分别是6.0和5.6;M2的半衰期是37 min,是M1的6.17倍;M2的比酶活是582.204U/mg,是M1的1.6倍.动力学分析显示,以普鲁兰糖为底物时,M2的Vmax、Kcat、Km和Kcat/Km分别为0.001 3μmoL/(mL·s)-1、191.80-1 s.-1、0.30 mg/mL、693.30,与M1相比,M2的底物亲和能力增加,同时催化效率是M1的2倍.本研究通过普鲁兰酶氮端氨基酸的截短,为提高酶的比酶活和半衰期提供了新的方法和思路.     

菜园土壤不同施肥模式下小青菜生长和品质及其生态化学计量学特征

利用盆栽试验研究了不同施肥处理对小青菜生长和品质的影响,并在此基础上利用生态化学计量学方法研究了小青菜生长和品质与其生态化学计量学特征之间的相关关系。结果表明在盆栽的条件下施氮量为180kg/hm2是小青菜生长较为合适的水平,能达到节氮保产的效果。在氮水平为180kg/hm2下,有机无机肥配施模式1(含有氨基酸肥料)提高了小青菜地上部的生长、碳浓度、光合速率、可溶性总糖、蔗糖、游离氨基酸含量和根系鲜重,且这些指标与其地上部C/N(碳氮比)和C/P(碳磷比)之间存在显著正相关关系,而与施肥模式中C/N和C/P之间存在正相关趋势,但相关关系不显著。此外,有机无机肥配施模式1降低了小青菜的地上部硝酸盐含量,且该指标与其地上部和施肥模式中N/P(氮磷比)、C/N、C/P和P/K(磷钾比)之间均不存在相关关系。因此,有机无机肥配施模式1促进了小青菜地上部和根系生长,提高了光合速率,改善了小青菜地上部的品质,是一种科学的施肥措施。     

Soil texture affects soil microbial and structural recovery during grassland restoration

Many biotic and abiotic factors influence recovery of soil communities following prolonged disturbance. We investigated the role of soil texture in the recovery of soil microbial community structure and changes in microbial stress, as indexed by phospholipid fatty acid (PLFA) profiles, using two chronosequences of grasslands restored from 0 to 19 years on silty clay loam and loamy fine sand soils in Nebraska, USA. All restorations were formerly cultivated fields seeded to native warm-season grasses through the USDA’s Conservation Reserve Program. Increases in many PLFA concentrations occurred across the silty clay loam chronosequence including total PLFA biomass, richness, fungi, arbuscular mycorrhizal fungi, Gram-positive bacteria, Gram-negative bacteria, and actinomycetes. Ratios of saturated:monounsaturated and iso:anteiso PLFAs decreased across the silty clay loam chronosequence indicating reduction in nutrient stress of the microbial community as grassland established. Multivariate analysis of entire PLFA profiles across the silty clay loam chronosequence showed recovery of microbial community structure on the trajectory toward native prairie. Conversely, no microbial groups exhibited a directional change across the loamy fine sand chronosequence. Changes in soil structure were also only observed across the silty clay loam chronosequence. Aggregate mean weighted diameter (MWD) exhibited an exponential rise to maximum resulting from an exponential rise to maximum in the proportion of large macroaggregates (>2000 μm) and exponential decay in microaggregates (<250 μm and >53 μm) and the silt and clay fraction (<53 μm). Across both chronosequences, MWD was highly correlated with total PLFA biomass and the biomass of many microbial groups. Strong correlations between many PLFA groups and the MWD of aggregates underscore the interdependence between the recovery of soil microbial communities and soil structure that may explain more variation than time for some soils (i.e., loamy fine sand). This study demonstrates that soil microbial responses to grassland restoration are modulated by soil texture with implications for estimating the true capacity of restoration efforts to rehabilitate ecosystem functions.     

农林废弃物超低酸水解装置及试验研究

自行设计了木质纤维素类生物质超低酸水解装置,该装置可以间歇进料方式实现生物质的超低酸水解。在该装置上分别以木聚糖和定量滤纸为模化物得到了半纤维素和纤维素水解的最佳工况,进而对白松、速生杨和玉米秸秆这三种在我国分布较为广泛的原料进行了超低酸两步水解研究,分别得到了51.24%、78.27%和84.10%的可水解原料转化率和48.19%、57.96%和37.48%总还原糖得率,并以GCMS定性还原糖外的其他产物,最后对水解残渣做了分析研究。     

Phytochemical and antioxidant properties of anthocyanin-rich Morus nigra and Morus rubra fruits

In this study, phytochemical and antioxidant properties of anthocyanin-rich mulberry species of Morus nigra L. (black mulberry) and Morus rubra L. (red mulberry) fruits harvested from across Turkey were investigated. Fruit color, total phenolics (TP), total monomeric anthocyanin (TMA), titratable acidity (TA), and individual sugar and organic acid compositions were determined. Total antioxidant capacity (TAC) of fruits was assessed by both the trolox-equivalent antioxidant capacity (TEAC) and the ferric reducing antioxidant power (FRAP) assays. Black mulberry exhibited higher TP, TMA, TAC and TA when compared to red mulberry. The average TP contents of M. nigra and M. rubra were 2737 and 1603 μg gallic acid equivalent in g fresh weight basis (GAE/g fw), respectively. M. nigra had the richest amount of anthocyanin with an average of 571 μg cy-3-glu/g fw. Overall, TAC averaged 10.5 and 12.0 mmol TE/L by the TEAC and FRAP methods, respectively. We found that FRAP, TEAC, TP and TMA were significantly correlated (r = 0.64–0.99) with each other. Fructose (5.27 g/100 mL) and glucose (5.81 g/100 mL) were determined to be the major sugars in both mulberries. M. nigra displayed a higher TA (2.05 g/100 mL) than M. rubra (0.78 g/100 mL), with citric acid as the major acid.     

Effects of fruit bagging on coloring and related physiology,and qualities of red Chinese sand pears during fruit maturation

Red Chinese sand pears (Pyrus pyrifolia Nakai) are particular to China. In order to determine the effects of fruit bagging treatments (including bag types, bag removal patterns and dates) on fruit qualities and to understand the mechanism of coloring of red Chinese sand pears, two experiments were carried out. In the first experiment, fruit of ‘Meirensu’ were firstly covered by light-impermeable paper bags with different levels of light permeable liners during their early development stage, then, the whole bag were/not removed or only the outer layer of bags were removed 3, 2 or 1 weeks before harvest. Thus, the fruit were/not totally re-exposed or were under different levels of sunlight transmission (80.31% or 34.71%). Non-bagged fruit were used as the control. Bagging treatments significantly affected the concentration of anthocyanin and the visual qualities of pear fruit. Compared to control, fruit re-exposed totally for 2 or 3 weeks accumulated the largest amount of anthocyanin and fruit receiving 80.31% and 34.71% of sunlight for 1–3 weeks could synthesize a little anthocyanin, indicating that high light intensity is imperative for coloring in red Chinese sand pears. Bagging treatments did not affect contents of total soluble sugars, but decreased organic acids contents in fruit. In the second experiment, fruit of ‘Meirensu’ and ‘Yunhongli No. 1’ were covered with only one type of light-impermeable bag during the early development stage and totally re-exposed after the bag removal 15 days before harvest. Fruit were then collected at different intervals to trace the time-course of coloring, and related physiology and inner qualities. With increasing time after the bag removal, the concentration of chlorophyll, carotenoid, flavonoid and total phenols changed little, but the concentration of anthocyanin accumulated extremely fast within 10 days after the bag removal in both cultivars and thereafter kept constant. ‘Yunhongli No. 1’ had higher anthocyanin contents and lower hue angle than ‘Meirensu’, indicating a higher potential of anthocyanin synthesis. After the bag removal, the sucrose contents and PAL activities increased gradually and correlation analysis revealed that they were highly correlated with anthocyanin accumulation in two cultivars. This study suggests that anthocyanin biosynthesis in red Chinese sand pears is a highly light dependent process and modified by genotypes. Based on the current results, in order to obtain red Chinese sand pear fruit with attractive appearance and good inner qualities, fruit must be covered with light-impermeable bags at the early stage of fruit development and the bag should be removed totally at least 10 days before harvest.