Age estimation of different tree species using a special kind of an electrically recording resistance drill

The determination of tree age is an important issue for urban green planning, forestry and dendrology; finding non-destructive and quasi-non-destructive methods for this purpose is of great theoretical and practical importance. The resistance drilling method is quasi-non-destructive because the average diameter of an opening that remains after drilling does not exceed 3 mm. Do electrically recording resistance drills allow precise assessment of tree age? The aim of this study is to assess the accuracy of determining the number of tree rings based on an examination of this special kind of drilling resistance profiles for three tree species, the pine Pinus sylvestris L., the oak Quercus robur L., and the birch Betula pendula ROTH. In 2015 and 2016, 15 pine trees, 15 oak trees, and 15 birch trees were randomly selected. For each studied tree, a measurement was conducted using the electrically recording resistance drill IML-Resi E400 with a flat-tipped 1.5/3 mm steel needle (research sample), and an increment core was taken (reference sample). The drill used was not a real Resistograph^®. The analysis of the E400-profiles underestimated the number of tree rings; the mean bias error (MBE) values were –6.5, –2.5, and –6.0 years for pine, oak, and birch, respectively. The proportion of investigated trees with less than five years difference between the research and reference samples varied from 38.4 (birch) to 66.7 (oak) percent. The accuracy of tree age determination was lowest for birch and highest for oak. The binomial generalised linear model (GLM) revealed that the most accurate tree age assessments were obtained from tree rings wider than 2 mm. The measurements clearly showed that the electrically recording resistance drill IML-Resi E400 enables a quick, although approximate, tree age assessment. Future research should concentrate on electronically regulating and recording drills, providing a higher spatial and signal resolution, and a stronger correlation to wood density.     

引种越桔果实中维生素E,SOD,氨基酸及无机元素含量

对从国外引种栽培的１０个越桔优良品种果实的成分分析表明，越桔果实中维生素Ｅ含量和超氧化物歧化酶（ＳＯＤ）活性较高，分别在２．７～９．５μｇ／ｇ和（５．３９～５５．３９）×１６．６７×１０－９ｍｏｌ／ｓ；果实中无机元素Ｃａ、Ｆｅ、Ｍｎ、Ｐ、Ｚｎ含量较丰富；果实中含有１５种氨基酸，总氨基酸含量在１．０８５％～２．５４９％之间。不同品种之间，果实中营养成分含量差异较大。综合评价认为，越桔果实为一优良的名优稀特果品。     

油料作物维生素E含量的研究进展

介绍了维生素E的类型、来源及理化性质;概述了生育酚在油料作物中的含量、组成及遗传变异研究进展和生育酚相关基因的研究动态;展望了提高油料作物维生素E含量的发展前景。     

低温诱导堆型艾美耳球虫3-1E基因的可溶性表达

设计合成特异引物应用RT-PCR技术扩增堆型艾美耳球虫的3-1E基因。通过EcoRⅠ和NotⅠ双酶切,基因连接到表达载体PET28a上,转化入Escherichia coli BL21(DE3)感受态细菌,挑取阳性菌落。经过双酶切和测序鉴定,证明含目的基因的表达载体PET28a构建成功。低温(16℃)诱导表达,经过SDS-PAGE和We-stren Bolt鉴定,表明此蛋白以可溶蛋白在细菌内表达。在诱导50h,IPTG浓度为0.4mmol/L时目的蛋白表达量最大。     

狂犬病病毒核蛋白基因在大肠杆菌中稳定表达条件的优化及纯化

将狂犬病病毒SRV9株核蛋白基因按正确的读码框克隆至GST融合表达载体pGEX-4T-1中,转化至大肠杆菌Rosetta株,IPTG诱导表达。表达的融合蛋白经SDS-PAGE分析显示,相对分子量约为82 kD,与预期大小一致。Western-blot检测结果表明,融合蛋白能与多克隆阳性血清发生特异性反应。为获取大量ELISA包被用核蛋白,试验还借助SDS-PAGE方法对重组目的基因的表达条件进行了优化,比较了诱导温度、菌密度I、PTG浓度、诱导时间等参数对重组基因表达的影响,以确定最佳诱导表达条件。结果表明:27~32℃,OD5500.3~0.4,IPTG 0.06 mmol/L、诱导至OD550值不再增加时为最佳诱导表达条件。优化后经扫描分析显示,所表达融合蛋白占菌体总蛋白的20%以上。经包涵体纯化和亲和层析纯化,可获得纯度较高的GST融合蛋白。     

可可泛素结合酶基因家族的生物信息学初步分析

泛素结合酶(E2s)促进底物泛素化或者与E3s链接,是靶蛋白泛素化的关键酶,在泛素-蛋白酶体途径中起重要作用。利用可可(Theobroma cacao L.)全基因组测序数据,共鉴定出45个E2s基因家族成员,包括39个UBC和6个UEV基因。通过生物信息学方法,对可可E2s家族的基本理化性质、基因结构、二级结构预测、亚细胞定位、进化关系等方面进行初步分析。结果表明:可可E2s基因CDS长度在441(Tc UEV3)~3 651 bp(Tc UBC7)之间,对应的编码蛋白氨基酸数目在146(Tc UEV3)~1 216 aa(Tc UBC7)之间,编码蛋白分子量在16.39~134.71 ku之间。E2s基因外显子在1~11之间,多数基因外显子数目在5~7之间。E2s基因在10条染色体上均有分布,1号染色体为7个,数量最多;6号染色体2个基因,数量最少。可可E2s蛋白大多为不稳定蛋白,且均为亲水性蛋白,其二级结构以α-螺旋和无规则卷曲为主要构成元件。大多数可可E2s蛋白定位在细胞核,少数定位在内质网或者细胞质。进化树分析表明:可可E2s蛋白被分为20个亚家族,包括16个UBC亚家族和4个UEV亚家族,第Ⅵ亚家族E2s数目最多为9个;E2s家族蛋白在物种进化过程中具有高度保守性。     

逆转录病毒载体介导的猪瘟病毒E2基因在真核细胞中表达及动物免疫试验

利用DNA重组技术将猪瘟病毒(CSFV)石门株囊膜蛋白E2基因插入逆转录病毒载体pBABE-puro 中构建成重组逆转录病毒载体pBABE-puro-E2,该重组逆转录病毒表达载体与pVSVg质粒经磷酸钙共转染法将其转入293GP细胞中包装逆转录病毒假病毒。用包装的假病毒感染SP2/0细胞,经嘌呤霉素筛选阳性细胞后进行流式细胞技术(FACS)分析,结果表明CSFV E2基因在SP2/0细胞膜上成功表达。将表达E2蛋白的SP2/0细胞腹腔免疫BALB/c小鼠,用流式细胞仪检测证明,成功诱导小鼠产生了抗E2蛋白的抗体。为下一步利用细胞免疫小鼠研制抗猪瘟病毒单克隆抗体打下基础。     

Genomic and Pathogenic Studies on a Glycoprotein E Variant Field Isolate of Bovine Herpesvirus 1

Glycoprotein E-negative (gE–) laboratory strains of bovine herpesvirus 1 (BHV-1) were recently introduced as novel marker vaccines, allowing serological discrimination between vaccinated and naturally infected animals on the basis of lack or presence of antibodies against gE epitopes. The applicability of this approach is based on the genetic stability of the gE. However, mutant field variants of BHV-1 with a variable response in anti-gE ELISA have been isolated. The molecular characterization of a gE variant field isolate (Salwa strain) is presented here. By comparing the gE nucleotide and amino acid sequences of the Salwa strain with those of the wild strain Jura, ten mutated bases were found in the gE strain of Salwa, six of which alter the amino acid sequence, leading to changes in five amino acids. Both strains caused respiratory disease in experimentally infected calves, but Salwa generated slightly milder signs. Both viruses were excreted in nasal and ocular discharges, and were reactivated by dexamethasone treatment. In conclusion, the rather close similarities observed in the gE gene structure and pathogenicity features of the gE mutant and of the wild strain of BHV-1 confirm the genetic stability of gE. The findings indicate that the Salwa isolate is virulent, but less virulent than wild strains. Our data support the use of gE-negative marker vaccines in eradication programmes.     

鸭大肠杆菌超广谱β-内酰胺酶的检测及药物敏感性分析

分析鸭大肠杆菌的耐药性与超广谱β-内酰胺酶的关系,为临床相关感染的治疗提供理论依据。采用CLSI推荐的初步筛选试验和表型确证试验方法,检测了临床分离的12株鸭大肠杆菌产超广谱β-内酰胺酶的情况,用微量稀释法测定了环丙沙星等9种抗菌药对其的抗菌活性。临床分离的12株鸭大肠杆菌中,有4株产超广谱β-内酰胺酶,检出率为33%。产超广谱β-内酰胺酶的菌株对多种药物耐药,对氟喹诺酮类药物之间产生交叉耐药,对氟喹诺酮类、氨基糖苷类、头孢菌素类产生多重耐药性,产酶菌的耐药性明显大于非产酶菌。产超广谱β-内酰胺酶是鸭大肠杆菌对抗菌药物产生耐药性的主要机制之一。     

淡紫拟青霉E7菌株固体放大发酵产孢培养基和工艺条件研究

为探讨淡紫拟青霉E7菌株固体放大发酵因子间的关系以获得大量孢子用于田间试验,通过正交试验设计优化固体发酵培养基成分和培养条件组合,并测定外加碳源、氮源、无机盐和装料方式对产孢量的影响。结果表明：以玉米粉+甘蔗渣+麸皮+壳聚糖组成的正交2号配方为最佳复合培养基质,分生孢子产量达7.13×109个/g,以发酵液pH+液相发酵终点+温度+初始含水量组成的正交4号配方为优适培养条件,分生孢子产量达8.97×109个/g;该菌株在优化后的培养基配方中添加0.4％蔗糖、0.2％蛋白胨、0.002％硫酸锰,以双层纱布袋装料按优化后的培养条件放大培养,产孢量最高可达到8.22×1010个/g。优化后的E7菌株发酵产孢培养基基质用量少,发酵成本低,适合于固体放大发酵生产淡紫拟青霉孢子。