过氧化物酶体增殖剂对稻瘟病菌生长发育及致病性的影响

【目的】探索过氧化物酶体增殖剂(Peroxisome proliferators,PPs)对稻瘟病菌生长发育及致病性的影响。【方法】在6种不同的PPs诱导下,观察比较稻瘟病菌过氧化物酶体数量及相关基因表达、生长速率、孢子萌发、附着胞形成与致病性的差异。【结果】在不同的PPs诱导下,稻瘟病菌过氧化物酶体数量均呈现明显的增加,同时过氧化物酶体形成相关基因PEX8、PEX11、PEX14的表达量升高;PPs影响病菌菌丝生长、分生孢子萌发及附着孢形成,并导致致病性的减弱。其中,2,4-D与阿司匹林的抑制效果最为显著。同时,2,4-D与ASA对稻瘟病菌过氧化物酶体形成突变体Δpex5和Δpex7的生长抑制效果与野生菌株相比明显增加。【结论】首次将PPs类化合物用于模式丝状病原真菌稻瘟病菌的研究。研究发现6种PPs均能够引起过氧化物酶体的增殖,并可抑制稻瘟病菌生长发育,降低致病性。     

恒定磁场出生前暴露对仔鼠生长发育的影响

目的:探讨不同强度恒定磁场出生前暴露对仔鼠生后生长发育的影响。方法:将30只孕鼠自怀孕第0天始分别置于不同强度的恒定磁场(0.04T;0.08T;0.12T)中饲养,10只孕鼠作为对照。孕鼠让其自然分娩。仔鼠生后第2、3、8、20、22天分别开始观察和记录耳廓张开、平面翻正、张眼、睾丸下降、阴唇张开的时间。结果:耳廓张开及睁眼的时间在0.12T组分别为4.42±0.48天及14.19±0.94天,较对照组耳廓张开时间4.01±0.47天和睁眼时间13.21±0.79天相比差异有统计学意义,与其他强度磁场组相比无显著性差异.平面翻正、阴唇张开及睾丸下降出现时间在各组间相比差异无统计学意义。结论:恒定磁场出生前暴露可能对仔鼠的生长发育起到一定的影响,这种影响可能与磁场强度具有相关性。随着磁场强度的增加各个器官生长发育的时间相对延长。     

Site-directed mutagenesis by combination of homologous recombination and DpnI digestion of the plasmid template in Escherichia coli

A rapid site-directed mutagenesis strategy using homologous recombination and DpnI digestion of the template in Escherichia coli is described. Briefly, inverse polymerase chain reaction amplification of the entire circular plasmid was performed by mutagenic primers with overlapping sequences ( approximately 15 bp) for generating PCR products with approximately 15 bp of homology on the terminal ends. On direct transformation of the amplified PCR products into restriction endonuclease DpnI-expressing E. coli BUNDpnI, homologous recombination occurs in E. coli while the original templates are removed via DpnI digestion in vivo, thus yielding clones harboring mutated circular plasmids. Nearly 100% efficiency was attained when this strategy was used to modify DNA sequences.     

酶联免疫吸附技术(ELISA)对大豆根瘤菌的鉴定

本文用直接ELISA法检测大豆根瘤菌USDA 110和RTt 50的纯培养菌体和根瘤。确定了该试验的最佳工作条件:酶标结合物HRP—Ab 110和HRP—Ab50的工作稀释度分别为1:3200和1:800,抗体Ab 110和Ab 50的工作稀释度分别为1:3200和1:800,抗原USDA 110和RTt 50的最适工作浓度均为6×10~7细胞/ml。该法能够特异地检测和区别慢生型和快生型大豆根瘤菌。在这两种类型的大豆根瘤菌中,同种内的少数菌株存在交叉反应,通过吸收可以消除,从而使ELISA的检测达到菌株     

假单胞菌Pseudomonas fluorescens Pf0-1中转录调控因子SpfB负调控DNA磷硫酰化修饰

[目的]DNA磷硫酰化修饰是DNA骨架上非桥接的氧原子以序列选择性和R-构型被硫取代的一种新型DNA修饰。目前,磷硫酰化修饰在多种细菌、古生菌以及人类致病菌中多有发现,但其分子调控机制尚不清楚。为了全面解析磷硫酰化修饰的调控机制,本文选择荧光假单胞菌Pf0-1为研究对象,开展了其DNA磷硫酰化修饰的调控机制研究。[方法]首先,构建了spfB基因缺失和回补菌株,使用碘能特异性断裂磷硫酰化修饰DNA的方法,研究了该基因缺失对修饰表型的影响。利用cDNA在相邻同方向的基因间隔区进行PCR,确定了磷硫酰化修饰基因簇spfBCDE内的共转录单元。通过荧光定量RT-PCR,分析了spfB基因缺失突变株中磷硫酰化修饰基因的转录量。利用异源表达并纯化得到的重组蛋白SpfB进行了体外功能研究。通过EMSA实验,验证了SpfB蛋白具有与spfB启动子序列结合活性。通过DNase I footprinting实验,精确定位了SpfB蛋白与DNA结合序列。[结果]spfB基因的缺失加剧了磷硫酰化修饰DNA断裂所致电泳条带弥散的表型,spfB基因的回补能够恢复该表型,证明spfB基因负调控磷硫酰化修饰。鉴定了spf基因簇中只含有1个共转录单元,且该共转录单元在△spfB突变株中转录水平明显上升。通过EMSA和DNase I footprint实验,检测了SpfB蛋白与磷硫酰化修饰基因spfBCDE的启动子区域5''-TGTTTGT-3''相结合。[结论]SpfB作为转录调控因子负调控磷硫酰化修饰基因spfBCDE的表达,为解析磷硫酰化修饰的调控机制和全面理解基因组上的部分修饰特征奠定了基础。     

Pedoecological effects of a sand-fixing poplar (Populus simonii Carr.) forest in a desertified sandy land of Inner Mongolia, China

To assess the pedoecological effects of a 23-year old poplar ( Populus simonii Carr.) forest on soil amelioration and vegetation restoration via soil erosion reduction and atmospheric dust retention in a desertified sandy land ecosystem, daily dynamics of wind speed, sand transport and dust deposition rates were monitored over an erosive period from April through June in 2001, using fixed observation sites located at different positions within and around the forest. Soil and vegetation characteristics at these sites were also measured. The observation sites were placed at distances of 15H (as control), 6H and 3H (H is mean tree height) from the forest edge of the windward side (abbreviated CK, 6H-W and 3H-W respectively), forest center (FC), and at distances of 0H, 6H and 8H from the forest edge of the leeward side (FE-L, 6H-L and 8H-L respectively). Daily mean wind speed was significantly lower in different observation sites than CK, with FC having the greatest reduction of wind speed and 6H-W the least reduction. Daily transport rate of sand by wind was also significantly lower in different observation sites than CK, with FE-L having the greatest reduction of wind erosion and 6H-W the least reduction. The fact that the poplar forest will lose its functions against wind at a distance of about 12-fold tree height from the forest edge of the leeward side suggests that the effective wind-preventing range of the poplar forest is about 150 m. There was marked spatial and seasonal distribution of dust-fall rate. Over space, the rate of dust-fall was much greater within the forest than outside the forest. Over time, the daily dust deposition rate was greatest in April, followed in decreasing order by May, June, July, September and August, closely linked to the seasonal distribution pattern of dust storm. Significant positive changes in soil and vegetation parameters of the different observation sites during the 23 years that the poplar forest was established suggest the perceptible pedoecological effects of the poplar forest on soil development and restorative succession of plant community within its immediate vicinity through windbreak, soil erosion reduction and atmospheric dust retention. Understanding these pedoecological effects may aid in the design of protective forest systems in arid and semi-arid areas.     

蜂毒溶血肽前体蛋白cDNA的克隆及其融合蛋白的表达

从蜜蜂毒腺中提取总RNA ,通过RT PCR扩增得到了蜂毒溶血肽前体蛋白的cDNA ,将扩增产物克隆到 pT7Blu T载体上 ,再进一步将插入片段酶切并连接到 pUC1 1 8载体上 ,构建了重组质粒pUMP。DNA序列分析结果表明 ,克隆得到的cDNA序列与所发表序列完全相同 ,且与 β 半乳糖苷酶部分序列构成正确的读码框。含重组质粒 pUMP的大肠杆菌DH5α表达了与β 半乳糖苷酶部分序列融合的蜂毒溶血肽前体蛋白     

甲壳素脱乙酰酶的研究概况及展望

甲壳素脱乙酰酶(chitin deacetylase,CDA,E.C.3.5.1.41)是一种能催化脱去甲壳素分子中N-乙酰葡糖胺链上的乙酰基,使之变成壳聚糖的酶。而壳聚糖因其独特的性质被广泛应用于医药、食品、化工、化妆品等行业。对CDA的来源、分离纯化和酶学性质、结构和催化机制、基因的克隆表达及应用前景等方面的研究进行了综述,并分析出今后的主要研究方向应在CDA基因的克隆表达、CDA底物的改造及CDA的结构和催化机制等方面。     

Methylation protects miRNAs and siRNAs from a 3'-end uridylation activity in Arabidopsis

Small RNAs of 21-25 nucleotides (nt), including small interfering RNAs (siRNAs) and microRNAs (miRNAs), act as guide RNAs to silence target-gene expression in a sequence-specific manner. In addition to a Dicer homolog, DCL1, the biogenesis of miRNAs in Arabidopsis requires another protein, HEN1. miRNAs are reduced in abundance and increased in size in hen1 mutants. We found that HEN1 is a miRNA methyltransferase that adds a methyl group to the 3'-most nucleotide of miRNAs, but the role of miRNA methylation was unknown. Here, we show that siRNAs from sense transgenes, hairpin transgenes, and transposons or repeat sequences, as well as a new class of siRNAs known as trans-acting siRNAs, are also methylated in vivo by HEN1. In addition, we show that the size increase of small RNAs in the hen1-1 mutant is due to the addition of one to five U residues to the 3' ends of the small RNAs. Therefore, a novel uridylation activity targets the 3' ends of unmethylated miRNAs and siRNAs in hen1 mutants. We conclude that 3'-end methylation is a common step in miRNA and siRNA metabolism and likely protects the 3' ends of the small RNAs from the uridylation activity.     

不同品种和生长年限对丹参产量及有效成分积累动态的影响

同时提高丹参经济产量(根系干重)及有效成分积累是当前丹参栽培的热点及难点。通过不同品种和生长年限丹参的田间小区栽培试验,测定不同采收期丹参根系干重,脂溶性成分含量(丹参酮I,隐丹参酮和丹参酮IIA)和水溶性成分含量(原儿茶醛,原儿茶酸,丹酚酸B和迷迭香酸),研究品种、生长年限、采收期对丹参经济产量和有效成分积累的影响,以期为丹参规范化种植提供依据。无论从经济产量,还是脂溶性成分和水溶性成分产量(根系产量×含量),两年生丹参均优于第一年生。丹参脂溶性成分含量在9月份达到最高,而水溶性成分含量除迷迭香酸外在11月份达到最高值。由于收获前丹参根系的快速生长,无论是水溶性还是脂溶性成分,其有效成分产量均在收获期(11月4日)达到最高。不同品种丹参品种的经济产量及有效成分存在较大差异,生产中应有针对性的选择丹参品种。