燕山板栗种质资源AFLP遗传多样性分析

为研究燕山板栗的遗传多样性,用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)技术对44份板栗种质的遗传多样性及遗传距离进行分析,其中36份原产燕山地区的种质中有18份为野生种质和18份为栽培品种,另外8份为原产山东的品种.共检测了384个AFLP位点,有217个是多态性位点,占56.5%,这表明了所检测的板栗材料中存在着明显的遗传多样性.用NT.SYS软件对44份板栗种质进行聚类分析.所有材料聚成4个类群,其中燕山地区原产的板栗材料被聚为3个类群,山东起源的被聚类为1个独立的类群.在所分析的44份材料中,遗传距离在0.0292和0.2214之间.燕山板栗和外来板栗存在着明显的差异,本研究结果与本实验室用RAPD技术分析的结果是一致的.结果对这些板栗材料在育种中的应用具有重要指导意义.     

葡萄果实cDNA文库的构建及鉴定

构建葡萄果实cDNA文库是研究果实发育相关基因表达调控的基础工作.本实验从巨峰葡萄果实中提取了高质量的RNA,利用MMLV反转录酶把总RNA中的mRNA反转录成cDNA第一链,用特异引物进行LD-PCR扩增合成双链cDNA.将分级纯化的带有粘性末端双链cDNA与λTrip Ⅰ Ex2载体连接,重组载体体外包埋成为cDNA原始文库.初步鉴定原始文库的滴度为1.32×106 pfu/mL,文库扩增后滴度达1.45×1010 pfu/mL.从扩增文库中随机挑取250个克隆进行PCR鉴定,结果表明重组率为98.7%,插入片段大小在0.5～2.0 kb之间,因此,本研究成功构建了葡萄果实cDNA文库.     

分子标记辅助选育小麦抗白粉病、优质高分子量麦谷蛋白亚基聚合体

现代小麦育种的目标是选育丰产、综合抗逆性好的优质小麦新品种,将分子标记技术与传统育种方法相结合可以大大提高育种效率.本研究利用与小麦抗白粉病基因Pm21紧密连锁的共显性PCR标记、以及优质高分子量麦谷蛋白亚基1Dx5 1Dy10特异的PCR标记对以小麦品种安农94212、安农92484为优质亲本和以抗白粉病小麦簇毛麦易位系为抗病亲本的杂交高代材料进行标记位点的检测,结合田间抗病性鉴定结果,筛选、培育出聚合有Pm21和1Dx5 1Dy10的抗白粉病小麦聚合体,为小麦抗病、优质育种提供了具有重要利用价值的中间材料.     

大花蕙兰幼苗叶片诱导类原球茎

以大花蕙兰无菌幼苗叶片为外植体,成功诱导类原球茎(PLB).叶片部位是影响诱导的关键,只有叶片基部片段能成功诱导出PLB.不同激素配比以及外植体来源对PLB诱导有重要的影响.低浓度的2,4-D和较高浓度的6-BA对于拟球茎诱导PLB具有显著的促进作用.当2,4-D浓度高于0.6 mg/L时,PLB诱导受到显著抑制.较高的2,4-D水平可能有利于PLB诱导初始阶段,但不利于PLB诱导中后期的进一步发育.不同基因型有不同的最佳激素组合.添加0.2 mg/L的NAA对大花蕙兰PLB诱导没有显著影响,但在有的基因型中对PLB诱导有促进作用.以叶片基部片段诱导大花蕙兰PLB是可行的,为转基因研究提供了再生技术基础.     

OsI2基因的克隆及其植物表达载体的构建

在旱稻IRAT109干旱胁迫下cDNA芯片分析结果的基础上,通过RT-PCR,5'-RACE及3'-RACE方法,从IRAT109总RNA中扩增得到了干旱胁迫下芯片表达谱中上升表达强度第二的基因全长序列,命名为OsI2,全长有523 bp,并对基因序列结构进行了分析.该基因与全长cDNA文库中的CT836140.1有99%同源.OsI2基因编码产物对应1个包含57个氨基酸的开放阅读框(ORF),为一功能未知蛋白.其编码产物也可能对应两个中间相隔19 bp的较小ORFs(43个氨基酸和42个氨基酸),都是功能未知的蛋白.在pBI121载体的基础上,将OsI2基因与CaMV35S连接成功构建了pBl121-OsI2植物表达载体,为进一步研究其功能创造了条件.     

南方红豆杉扦插繁殖技术研究

2003~2004年应用2年生南方红豆杉幼苗进行扦插试验研究,结果表明:不同时间扦插的插穗生根时间最多相差2个月,2月中旬至3月中旬为最适宜扦插季节,扦插后3~5个月的生根率高于其它月份。用河沙与红泥作基质扦插,在不同季节扦插中以河沙作基质的生根效果比红泥的好。用生根剂处理有利于插穗提早生根及增加生根数量,以冬季及早春应用的效果较为明显,应用生根剂的插穗在扦插3~5个月后的生根率和生根数量与对照比有显著差异。不同类型插穗的扦插生根率在不同月份扦插的结果不同,但其差异不显著。     

In Vitro and In Vivo Dendritic Cell Immune Stimulation Effect of Low Molecular Weight Fucoidan from New Zealand Undaria pinnatifida

Low molecular weight fucoidan (LMWF) has been reported to have immunomodulation effects through the increase of the activation and function of macrophages. In this study, the regulating effect of LMWF from Undaria pinnatifida grown in New Zealand on dendritic cells (DCs) was investigated. We discovered that LMWF could stimulate DCs’ maturation and migration, as well as CD4+ and CD8+ T cells’ proliferation in vitro. We proved that this immune promoting activity is activated through TLR4 and its downstream MAPK and NF–κB signaling pathways. Further in vivo (mouse model) investigation showed that LMWF has a strong immunological boosting effect, such as facilitating the proliferation of immune cells and increasing the index of immune organs. These findings suggest that LMWF has a positive immunomodulatory effect and is a promising candidate to supplement cancer immunotherapy.     

Complete Genome Sequencing and Analysis of Rehmannia Mosaic Virus Isolate from Shanxi Province

Using double-stranded RNA(dsRNA) technology and sequence-independent amplification(SIA), the molecular identification on infected Rehmannia glutinosa in the field with mosaic symptoms was performed and the whole-genome of the Rehmannia mosaic virus(ReMV) Shanxi isolate(ReMV-SX) was sequenced. Sequencing analysis showed that the virus that infected Rehmannia glutinosa was Rehmannia mosaic virus(ReMV). The full-length of the obtained ReMV-SX sequence(GenBank accession no. JX575184) was 6 395 nt, containing four open reading frames(ORFs). The sequence homology analysis of the complete nucleotide sequence showed that ReMV-SX was 93.8%-97.0% homologous to ReMV in Tobamovirus subgroup I, while only 49.8%-58.9% homologous to the isolates in subgroups II and III of the same genus. Phylogenetic analysis showed that ReMV-SX and ReMV-Henan formed a separate branch and had the closest genetic relationship. The results laid the foundation for ongoing researches in the taxonomic status and evolution of ReMV and for further investigating the pathogenic mechanism of ReMV infecting Rehmannia glutinosa.     

Rare β-Resorcylic Acid Derivatives from a Halophyte-Associated Fungus Colletotrichum gloeosporioides JS0419 and Their Antifungal Activities

Six new β-resorcylic acid derivatives (1–5 and 7) were isolated from a halophyte-associated fungus, Colletotrichum gloeosporioides JS0419, together with four previously reported β-resorcylic acid lactones (RALs). The relative and absolute stereochemistry of 1 was completely established by a combination of spectroscopic data and chemical reactions. The structures of the isolated compounds were elucidated by analysis of HRMS and NMR data. Notably, compounds 1–3 had a β-resorcylic acid harboring a long unesterified aliphatic side chain, whereas the long aliphatic chains were esterified to form macrolactones in 4–9. Among the isolated compounds, monocillin I and radicicol showed potent antifungal activities against Cryptococcus neoformans, comparable to clinically available antifungal agents and radicicol showed weak antifungal activity against Candida albicans. These findings provide insight into the chemical diversity of fungal RAL-type compounds and their pharmacological potential.     

水分胁迫下不同抗旱类型大豆叶片气孔特性变化研究

采用火棉胶印膜法,研究了不同程度干旱胁迫对不同抗旱类型大豆叶片气孔特性的影响.结果表明:经水分胁迫的大豆叶片气孔密度增加,气孔开口大小和单位叶面积气孔相对面积减小.但不同抗旱类型大豆品种间表现不同,气孔密度变化幅度大小顺序依次为:弱抗品种(A3)＞中抗品种(A2)＞抗旱品种(A1),调节气孔开张能力大小顺序依次为:抗旱品种(A1)＞中抗品种(A2)＞弱抗品种(A3).