杉木木质化过程特异表达基因文库的构建与分析

研究木质部木质化过程及其生物学特性对于了解木本植物水分和矿物质的吸收及运输机制、木材形成的遗传控制、改良材性及提高木材生长量具有十分重要的意义。为了获得杉木木质化过程中特异表达的基因,本研究利用抑制差减杂交技术,以杉木突变体独干杉无性系为测试方(Tester),正常的句容0号无性系为驱动方(Driver),构建了正向和反向两个差减文库,分别获得了618和409个克隆。利用通用引物T7和SP6进行PCR及EcoRⅠ酶切鉴定文库重组子,结果证明所构建的文库符合要求,具有代表性。该文库的成功构建为获得杉木木质化过程中特异表达的基因及深入了解木本植物木质化的机理奠定了基础。     

东方百合鳞茎快速增长的组培体系研究

以东方百合栽培品种‘Sorbonne’和‘Siberia’的组培苗为试验材料,研究了蔗糖、6-BA、IAA、PP333及活性炭（AC）的添加浓度对组培苗小鳞茎增重和直径生长的影响;结果表明：80g／L蔗糖浓度有利于‘Sorbonne’和‘Siberia’组培苗小鳞茎增重;6-BA浓度为0．2mg／L、IAA浓度为0．5和1．0mg／L时‘Sorbonne’和‘Siberia’组培苗小鳞茎平均直径增加最大,分别为120．7％和138．0％;PP333浓度达3．2mg／L和1．6mg／L时,‘Sorbonne’和‘Siberia’的组培苗小鳞茎平均直径增加最大,分别达到505．7％和211．3％;AC浓度达到2．0g／L时‘Sorbonne’试管苗小鳞茎平均直径增加最大,浓度达到4．0g／L‘Siberia’组培苗小鳞茎平均直径增加最大,分别为194．7％和220．3％。     

丛枝菌根真菌调控土壤氧化亚氮排放的机制

氮素是陆地生态系统初级生产力的主要限制因子,自Haber-Bosch反应以来,氮肥的生产和施用极大地提高了粮食产量.然而过量施用氮肥导致氮肥利用率低,并造成了严重的环境污染,包括氮沉降、硝态氮淋洗以及N2O排放等.微生物直接参与土壤氮素循环,固氮微生物、氨氧化和反硝化微生物分别在土壤固氮、铵态氮转化和硝态氮转化过程中起...     

Genomic footprints of wheat evolution in China reflected by a Wheat660K SNP array

Common wheat(Triticum aestiuum L.)is one of the most important crops because it provides about 20%of the total calories for humans.T.aestiuum is an excellent modern species for studying concerted evolution of sub-genomes in polyploid species,because of its large chromosome size and three well-known genome donors.Establishment of common wheat genome reference sequence and development of high-density SNP chips provide an excellent foundation to answer questions of wheat evolution and breeding at the genomic level.By genotyping more than 600 accessions of common wheat and their diploid and tetraploid ancestors using a Wheat660 K SNP array,we found dramatic genome changes due to tetraploidization and hexaploidization,in contrast to weaker influences of domestication and breeding on them.Further,since common wheat was introduced in China in 1500 BCE,Chinese landraces formed two subgroups(T.aestiuum-L1 and T.aestiuum-L2)with considerably diverse geographic distributions and agronomic traits.T.aestiuum-L2,mainly distributed in central and east China is found to have more but smaller oval grains with early maturity characteristics.We found that variation and selection in intergenic regions of the A and B sub-genomes dominated this differentiation,in which chromosomes 7 A and 3 B took the leading roles due to the existence of putative genes related to defense responses and environmental adaption in the highly differentiated regions.Large haplotype blocks were detected on 3 B(232.6-398.3 Mb)and 7 A(211.7-272.9 Mb)in the landraces,forming two distinct haplotypes,respectively.We discovered that artificial crosses in breeding promoted recombination in the whole genome,however,this recombination and differentiation was highly asymmetric among the three sub-genomes in homoeologous regions.In addition,we found that the wide use of European and northern American cultivars in breeding at early era,led dramatic changes in Chinese wheat genome,whereas,the recent breeding functioned to optimize it.This study will provide the insight for reconsideration of wheat evolution and breeding,and a new strategy for parent selection in breeding.     

Generation of a uniform thymic malignant lymphoma model with C57BL/6J p53 gene deficient mice

Lymphoma is the third most common cancer diagnosed in children, and T-cell lymphoma has the worst prognosis based on clinical observations. To date, a lymphoma model with uniform penetrance has not yet been developed. In this study, we generated a p53 deficient mouse model by targeting embryonic stem cells derived from a C57BL/6J mouse strain. Homozygous p53 deficient mice exhibited a higher rate of spontaneous tumorigenesis, with a high spontaneous occurrence rate (93.3%) of malignant lymphoma. Because tumor models with high phenotypic consistency are currently needed, we generated a lymphoma model by a single intraperitoneal injection of 37.5 or 75 mg/kg N-methyl-N-nitrosourea to p53 deficient mice. Lymphoma and retinal degeneration occurred in 100% of p53^+/− mice administered with higher concentrations of N-methyl-N-nitrosourea, a much greater response than those of previously reported models. The main anatomic sites of lymphoma were the thymus, spleen, bone marrow, and lymph nodes. Both induced and spontaneous lymphomas in the thymus and spleen stained positive for CD3 antigen, and flow cytometry detected positive CD4 and/or CD8 cells. Based on our observations and previous data, we hypothesize that mice with a B6 background are prone to lymphomagenesis.     

花鲈虹彩病毒交叉引物恒温扩增检测方法的建立

【目的】花鲈虹彩病毒(Lateolabrax maculatus iridovirus,LMIV)严重威胁花鲈养殖业安全,无特效防控药物,早期诊断在LMIV防控中发挥极其重要的作用。建立一种简便、快捷、准确的现场快速诊断方法,可为LMIV的基层诊断提供技术支撑。【方法】利用交叉引物恒温扩增技术(Cross priming amplification,CPA),针对LMIV ATPase基因高保守区设计1套单交叉引物。以构建的ATPase重组质粒作为阳性模板,对反应体系中的引物浓度比,Bst DNA聚合酶、Betaine、MgSO₄、dNTP浓度,以及反应温度和反应时间进行优化;结合一次性核酸试纸条,建立可视化检测LMIV-CPA的方法。【结果】最优引物浓度比组合为交叉引物CPF1.0μmol/L,引物F3和B3均为0.4μmol/L,探针引物B1(FAM)和B2(Biotin)均为0.8μmol/L;MgSO4浓度为6 mmol/L、Betaine浓度为0.4 mol/L、dNTP浓度为0.6 mmol/L、Bst DNA聚合酶浓度为0.256 U/μL;最佳反应...     

PBL模式《小动物透明标本制作与技术改良》建设与教学探索

《小动物透明标本制作与技术改良》课程引入国际流行的PBL(Problem based learning)教学模式,通过教师引导,学生自主学习、讨论制定实验方案,主动动手操作,并总结汇报实验成果的教学流程,探讨小动物透明标本的制作方法及技术改良手段,以及创新性实验课程的开设模式.教学实践表明:基于PBL的实验教学模式不仅...     

生物矿化合成木材-SiO2复合材料的XPS分析

利用生物矿化原理制备木材—SiO2复合材料,采用溶胶—凝胶法制备硅溶液,用不同质量分数的硅溶液对喜树进行浇灌和滴注试验。经过1个月的试验得到样品,利用光电子能谱仪(XPS)分析素材和SiO2复合材样品中硅、碳的质量比,证实喜树成功吸收硅元素,并且存活,植株根茎附近生长较好,枝叶较多,顶部附近枝叶较少。比较浇灌、滴注2种方法:①浇灌过程相对简单,树木对硅的吸收相对较少;使用质量分数越高的药品对树木进行浇灌,树木对硅的吸收越好。②滴注过程相对复杂,树木对硅的吸收相对较多;使用质量分数越高的药品进行滴注,树木对硅的吸收越好;但吸收不均匀,呈滴注孔口附近往树梢方向减少趋势。前期研究结果表明,对树木进行浇灌和滴注,药品硅的质量分数过高树木会死亡,过低则达不到要求,因此,最佳质量分数配制还有待于进一步研究确定。总的来说,利用生物矿化原理模拟木材—SiO复合材是可行的。     

一种基于边缘形状的边缘连接方法

在图像处理领域,边缘检测是一个重要的步骤,在数字图像分割、立体匹配、目标识别等领域里有着重要的作用。在检测出来的边缘中,有很多间断的部分,这使图像分割变得更加困难,为使分割更加理想,需要将间断部分连接起来。本文提出一种应用差分原理,基于边缘形状的边缘连接方法。通过对新方法进行理论分析和对比实验,此方法能有效连接间断边缘。     

桃蛀螟对青霉菌侵染苹果的产卵选择与行为趋向反应

为探明桃蛀螟Conogethes punctiferalis偏好于在青霉菌Penicillium侵染的苹果上产卵的特点与机理,从苹果、玉米和柑橘上分离、纯化获得15株青霉菌,通过形态学和rDNA-ITS序列分析鉴定后,以健康苹果为对照,测试不同青霉菌菌株侵染的苹果对桃蛀螟产卵选择的影响,并利用四臂嗅觉仪测试桃蛀螟对青霉菌诱导的苹果挥发物的行为趋向反应。结果表明,15株青霉菌分别属于皮落青霉P. crustosum、橘青霉P. citrinum、苏门答腊青霉P. sumatrense和指状青霉P. digitatum的不同株型;桃蛀螟对其中的12株青霉菌侵染的苹果的产卵选择率均极显著高于健康苹果,尤其对皮落青霉CO5菌株、苏门答腊青霉OR3菌株和指状青霉OR5菌株侵染的苹果的产卵选择率最高,分别达到67.73%、61.50%和68.39%;桃蛀螟对感染皮落青霉CO5菌株和指状青霉OR5菌株的苹果挥发物的选择率分别为31.37%和29.19%,均显著高于对感染橘青霉OR1菌株和苏门答腊青霉OR3菌株的苹果挥发物的选择率18.14%和21.31%,且前二者之间、后二者之间均无显著差异。表明青霉菌可通过影响寄主植物挥发物释放,从而影响桃蛀螟对寄主植物的产卵选择和行为趋向反应。