越来越苍老的世界

中国全面推行计划生育政策,少生了3亿多人,节约了7万亿抚养费,也少了3亿可以负担养老的劳动力。由于老龄化加速,老年抚养比由1964年的6.4%上升为2002年的11.6%,离退休金平均实际递增18.9%。到2050年,中国的抚养比将可能达到70%。老龄化速度,法国用了115年,英国用了80年,美国用了60年,我国用了18年。     

远程教育培训给生物教学带来新的生机

远程教育培训给生物教学带来新的生机。夯实了专业知识,梳理了生物知识,使我认识到应该如何把握生物课堂教学,提高了课堂教学设计能力,凝聚了生物教师的力量,使老师们学到了很多知识和教学技能,解决了平时教学中的一些难题,提升了生物的教学水平。     

亚热带丘陵茶园间作白三叶的土壤环境调控效果

在亚热带丘陵区1年生幼龄茶园通过连续4年间作白三叶大田试验,研究了其对茶园土壤环境及茶叶品质、产量的影响,结果表明,茶园间作白三叶降低了土壤容重,提高了土壤孔隙度、有机质、全氮、水解氮的含量及钾的活性,消耗了部分有效磷;增加了土壤关键层次（0～20cm）和关键时期（4～6月）的水分含量,延缓和缩短了夏季高温干旱与秋季持续干旱时间;获得了土壤降温时增温、保温与升温时降温的双向动态调控效果．增加了同一层次土壤温度的稳定性,延缓了高温和低温的出现时间,缩短了过度高温时间。从而促进了茶树生长,改善了茶叶品质,显著增加了茶叶产量。与清耕茶园相比,茶园间作白三叶后,春秋茶的酚氨比分别下降了17．10％和30．90％,产量提高了32.65％。     

酷想大晒台

哎呀呀,这个世界乱套了!大人变成了小孩,小孩变成了大人。现在,终于轮到各位小酷想家来当家做主了,快来看看他们都有什么出色的表现吧!看到爸妈变成了小孩,我立马学着他们的样子,下达了第一道命令:写作业去!我想他们一定会哭的,因为老师留的作业实在是太多了。新浪YOYO     

砷对烤烟(Nicotiana tabacum L.)碳代谢的影响

采用盆栽试验,系统地研究了砷对烤烟全生育期碳代谢及其过程的影响,结果表明,砷降低了烤烟整个生育期的叶绿素含量、光合速率、蔗糖合成酶（SS,合成方向）活性和现蕾以后的蔗糖磷酸合成酶（SPS）活性,提高了全生育期的SS（分解方向）活性和可溶性糖含量,因而抑制了碳的同化和蔗糖的合成,促进了蔗糖的分解,不利于碳向积累方向转化。砷提高了全生育期的腺苷二磷酸葡萄糖焦磷酸化酶（ADPG-PPase）活性,增加了团棵期和现蕾期淀粉的积累,降低了团棵期和采收期的可溶性淀粉酶（SSS）活性和采收期的淀粉含量,从而导致了碳积累代谢的紊乱,最终造成碳积累的减少。     

生态条件的多样性变化对蜜蜂生存的影响

蜜蜂在整个生态系统中起着重要的传花授粉作用,是生态链中不可或缺的物种。随着现代农业的发展,蜜蜂赖以生存的环境遭到破坏,继而引发蜜蜂数量大幅减少,影响了蜂种的生存与可持续发展。总结了近年来生态条件的变化,归纳了影响蜜蜂生存的主要因素,分析了蜜蜂生存艰难的原因,提出了蜜蜂生存的关键问题,并展望了未来维持蜜蜂强群的主要研究方向。     

一年又一年

上了年纪的人都有相似的感觉,就是年纪越大,时间好像过得越快。小时候,过了元旦盼春节,过了春节盼五一、六一,过了六一盼国庆,过了国庆再盼元旦。时间总是那么漫长,盼望的假期似乎总是姗姗来迟。而如今,似乎一眨眼,一年就过去了。我给这个现象总结了一个理论,叫做相对     

中国农业生物技术发展概况

当前在世界范围内由于人口的迅速增长导致的粮食危机正日益为各国科学家和政治家所关注。中国既是一个人口大国、也是个农业大国。半个世纪以来,特别是近十年来,在坚持四项基本原则和改革开放的方针指引下,我们以占世界7％的耕地养活了占世界近22％的人口,基本解决了温饱问题,并且提供了较充足的工业原料,取得了瞩目的成就。1984年后,虽然粮食产量出现了徘徊形势,但1989年,粮食产量有较大回升,达到了40475万吨,超过了1984     

促进微生物实验技术创新水平的改革初探

为了加强对本科生综合素质和能力的培养 ,激发学生的学习兴趣 ,我们对微生物实验课的教学内容进行了一系列改革 ,优化了实验项目 ,节约实验成本 ,并且引入科研成果转化实验 ,同时改进了实验方法和手段 ,实验结果更为准确。这些措施蕴含了较高的理论水平 ,增加了教学信息量 ,明显提高了教学效果 ,为培养高素质人才奠定了基础。     

转基因鸡的制备方法探讨

由于鸡本身的众多优点,转基因鸡的研究逐渐受到了研究者的重视,简要介绍了转基因鸡的发展史,对各种转基因技术方法进行了比较和分析,提出了每种方法的优缺点,尤其在精子介导法上作了较为详细的论述,并展望了转基因鸡的应用前景。     

Transposon Tn5 mutagenesis of Pseudomonas fluorescens to isolate mutants deficient in antibacterial activity

Abstract Pseudomonas fluorescens was subjected to insertion mutagenesis studies using the transposon Tn5-GM to generate mutants deficient in antibacterial activity minus mutants. The transposon located on the temperature-sensitive plasmid pCHR84 was conjugally transferred into the non-pathogenic pseudomonad using the triparental mating procedure. Random integration of Tn 5 -GM into the chromosome of P. fluorescens was achieved by heat ttreatment of the transformed cells at 42°C. Approximately 2% of transconjugants revealed an auxotrophic phenotype indicating efficient integration of the employed transposon into the chromosome of P. fluorescens . One transposon insertion mutant was obtained showing an antibacterial activity minus phenotype. This mutant (MM-7) was found to be defective in the production of an unidentified antibacterial compound against B. subtilis . These results introduce Tn 5 transposon mutagenesis as a new useful tool for the molecular analysis of P. fluorescens .     

The role of Pro-239 in the catalysis and heat stability of subtilisin E

Site-directed mutagenesis was employed to analyze the role of an alpha-helix containing catalytic Ser-221 of subtilisin E. Pro-239 located at the carboxy-terminal end of the alpha-helix was first replaced with Gly to examine the role of Pro-239 in the catalysis and stability of subtilisin E. The mutation was found to decrease both the catalytic rate (kcat) and the heat stability. This result strongly suggests that Pro-239 plays an important role in the maintenance of the alpha-helix, affecting the functioning of the active site. Various amino acid substitutions at position 239 were attempted to obtain the active subtilisins from Gly-239 subtilisin. Lys- and Arg-substitutions were found to result in more active and stable subtilisins than the Gly-239 subtilisin. In particular, the Arg-239 mutant showed enhanced heat stability compared with the wild type. These results demonstrate the important role of the alpha-helix containing catalytic Ser-221 in the catalysis as well as in the heat stability of subtilisin.     

Mutants of d-aminopeptidase with increased thermal stability

Mutant d-aminopeptidases from Ochrobactrum anthropi with increased thermal stability were obtained by random mutagenesis. One of the mutants, no. 65, was derived from E. coli cells transformed with DNA treated with sodium nitrite. The remaining activity of the purified mutant ezyme no. 65 after heat treatment at 52°C for 10 min was 20% that of the untreated mutant enzyme no. 65, whereas the native enzyme showed 5% of the untreated native enzyme activity after the same treatment. The gene for the mutant enzyme no. 65 was sequenced and it was found that Gly155 and Gly279 in the native enzyme were replaced by Ser and Asp, respectively. Five mutants carrying one or two mutations were generated from the native gene by site-specific mutagenesis. The enhancement of the thermal stability of mutant enzyme no. 65 was attributed to the substitution of Gly155 to Ser.     

Increased efficiency of targeted mutagenesis by CRISPR/Cas9 in plants using heat stress

The CRISPR/Cas9 system has greatly improved our ability to engineer targeted mutations in eukaryotic genomes. While CRISPR/Cas9 appears to work universally, the efficiency of targeted mutagenesis and the adverse generation of off‐target mutations vary greatly between different organisms. In this study, we report that Arabidopsis plants subjected to heat stress at 37°C show much higher frequencies of CRISPR‐induced mutations compared to plants grown continuously at the standard temperature (22°C). Using quantitative assays relying on green fluorescent protein (GFP) reporter genes, we found that targeted mutagenesis by CRISPR/Cas9 in Arabidopsis is increased by approximately 5‐fold in somatic tissues and up to 100‐fold in the germline upon heat treatment. This effect of temperature on the mutation rate is not limited to Arabidopsis, as we observed a similar increase in targeted mutations by CRISPR/Cas9 in Citrus plants exposed to heat stress at 37°C. In vitro assays demonstrate that Cas9 from Streptococcus pyogenes (SpCas9) is more active in creating double‐stranded DNA breaks at 37°C than at 22°C, thus indicating a potential contributing mechanism for the in vivo effect of temperature on CRISPR/Cas9. This study reveals the importance of temperature in modulating SpCas9 activity in eukaryotes, and provides a simple method to increase on‐target mutagenesis in plants using CRISPR/Cas9.     

Germline mutagenesis mediated by <Emphasis Type="Italic">Sleeping Beauty</Emphasis>transposon system in mice

Following the descovery of its transposition activity in mammalian culture systems, the Sleeping Beauty (SB) transposon has since been applied to achieve germline mutagenesis in mice. Initially, the transposition efficiency was found to be low in cultured systems, but its activity in germ cells was unexpectedly high. This difference in transposition efficiency was found to be largely dependent on chromosomal status of the host genomic DNA and transposon vector design. The SB transposon system has been found to be suitable for comprehensive mutagenesis in mice. Therefore, it is an effective tool as a forward genetics screen for tagged insertional mutagenesis in mice.     

Vinyl chloride dependent mutagensis: effects of liver extracts and free radicals.

The mutagenic effects of vinyl chloride (VC) on Salmonella typhimurium strain TA1530 are enhanced by mouse or rat liver extracts. The extracts prepared from mice pretreated either with vinyl chloride or the microsomal enzyme inducer, Aroclor 1254, did not produce any greater stimulation of VC-dependent mutagenesis than extracts from untreated animals. These same extracts, however, differed markedly in their capacity to stimulate the mutagenicity of dimethylnitrosamine (DMN), a compound which is converted to a mutagen by an NADPH dependent microsomal mixed function oxidase. The order of activity of the extracts with DMN was Aroclor pretreated is greater than untreated is greater than VC pretreated. Furthermore, the stimulatory effect of the liver extracts on VC mediated mutagenesis did not require NADPH and was still evident in liver extracts in which the microsomal mixed function oxidase system had been heat inactivated. The mutagenic activity of VC also was found to be stimulated by riboflavin in the presence of light suggesting that free radicals may be involved in VC dependent mutagenesis.     

Effects of heat shock on the induction of mutations by chemical mutagens in Neurospora crassa

Preheating of Neurospora conidia increased their susceptibility to mutation induction by chemical mutagens. Optimal conditions of heat shock for enhanced mutagenesis were determined in 2.5 X 10(7) conidia/ml 0.067 M KH2PO4-Na2HPO4 (pH 7.0) buffer to be treatment at 43 degrees C for 60 min. When protein synthesis during heat stock was eliminated by cycloheximide or by use of the temperature-sensitive mutation psi-1, induction of thermotolerance was inhibited while induction of the enhanced state of mutability was not. Therefore, inducible protein synthesis is not involved in this process. To discover whether DNA-repair systems are altered by heat shock and, as a result, whether reversion frequencies increase, DNA-repair mutants (upr-1, uvs-2, uvs-3, uvs-6, mus-7, mus-16) were heated and their reversion frequencies at the ad-8 locus were measured. All the DNA-repair mutants showed higher reversion frequencies with MNNG treatment after heat shock than in non-heated control. It therefore seems that DNA repair is not involved in the enhancement of chemical mutagenesis by heat shock. Heat shock does not increase frequencies of reversion induced by ultraviolet light, and heat shock after treatment with chemical mutagens does not affect reversion frequencies. These results suggest that heat shock may change the structure and function of cellular membranes and thereby increase the influx of mutagens into cells.     

An inducible targeted tagging system for localized saturation mutagenesis in Arabidopsis

We describe a system of inducible insertional mutagenesis based on the Ac-Ds family of transposons for targeted tagging in Arabidopsis (Arabidopsis thaliana). In this system, the Ac and Ds elements are carried within the same T-DNA and a heat shock-inducible transposase fusion is utilized to control the levels of transposase gene expression, generating transpositions that can be subsequently stabilized without requiring crossing or segregation. We have mapped 40 single-copy lines by thermal asymmetric interlaced-PCR, which can be used as potential launch pads for heat shock mutagenesis. Using a starter line selected for detailed analysis, the efficiency of tagging over a 50-kb region in the genome was examined. Hits were obtained in the targeted genes with multiple alleles for most genes, with approximately equal numbers of hits detected in genes on either side of the T-DNA. These results establish the feasibility of our approach for localized saturation mutagenesis in Arabidopsis. This system is very efficient and much less laborious as compared to conventional crossing schemes and may be generally applicable to other plant species for which large-scale T-DNA tagging is not currently feasible.     

Effect of the weak Ca(2+)-binding site of subtilisin J by site-directed mutagenesis on heat stability.

The functional role of the negatively charged amino acid residue in subtilisin J from Bacillus stearothermophilus has been investigated by site-directed mutagenesis. Glu-195 located at the weak Ca2+-binding site was replaced with Gln to examine the role of Glu-195 in the heat stability of subtilisin J. Mutant enzyme was expressed in Bacillus subtilis and was purified from the culture supernatant. When the mutant enzyme was expressed at 37 degrees C in the presence of 2mM calcium chloride, the pattern of enzyme production was quite different from that of wild-type. The purified Gln-195 mutant enzyme was analyzed with respect to optimal temperature, optimal pH, and heat stability. The mutation was found to decrease the heat stability but not catalytic efficiency (kcat/Km) and optimal pH. These results demonstrate the important role of the negatively charged side chains at the weak Ca(2+)-binding site in the heat stability of subtilisin.     

Coexpression of UmuD'' with UmuC suppresses the UV mutagenesis deficiency of groE mutants.

The GroE proteins of Escherichia coli are heat shock proteins which have also been shown to be molecular chaperone proteins. Our previous work has shown that the GroE proteins of E. coli are required for UV mutagenesis. This process requires the umuDC genes which are regulated by the SOS regulon. As part of the UV mutagenesis pathway, the product of the umuD gene, UmuD, is posttranslationally cleaved to yield the active form, UmuD'. In order to investigate what role the groE gene products play in UV mutagenesis, we measured UV mutagenesis in groE+ and groE strains which were expressing either the umuDC or umuD'C genes. We found that expression of umuD' instead of umuD will suppress the nonmutability conferred by the groE mutations. However, cleavage of UmuD to UmuD' is unaffected by mutations at the groE locus. Instead we found that the presence of UmuD' increased the stability of UmuC in groE strains. In addition, we obtained evidence which indicates that GroEL interacts directly with UmuC.