香蕉MaPRMT1基因的分离及表达分析(英文)

[Objective] The aim of experiment was to lay molecular foundation for studying maturity mechanism of banana after harvest. [Method] The combined method of suppressing subtractive hybridization and cDNA micro-array were used to obtain cDNA segment of one PRMT gene in banana and the whole cDNA sequence of the gene was cloned.The bioinformatics analysis was operated on it,in addition, the expression profile analysis was conducted in different organs and different mature periods of banana.[Result] The whole length of cDNA in MaPRMT1 was 1 158 bp and possessed a complete open reading frame,which could encode 385 amino acids.It had high homology with PRMT in plant,containing one Methyltransf₁ domain.The MaPRMT1 gene was expressed in root,stem,leaf and fruit of banana and the expression levels in stem and leaf were relatively high.As the increase of days after harvest,the expression level declined gradually,however it reached maximum when ethylene release was biggest,then it declined.[Conclusion] MaPRMT1 belonged to the first kind of arginine methyltransferase and it was expressed differently in different organs and fruits at different mature periods.     

Research on the Construction and Function of a Chlorophyll Degradation Recombinant Engineering Strain

In order to effectively reduce the chlorophyll content in flue-cured tobacco, improve the overall quality of tobacco leaves, chlorophyllase gene was cloned from Arabidopsis thaliana. After the expression of the expression vector in E. coli, the recombinant engineering strain was obtained. Afterwards, IPTG(isopropy-β-D-thiogalactopyranoside)was used to induce the goal protein, and the chlorophyllase activity of the recombinant engineering strain was measured, so as to investigate its degradation effect on the chlorophyll in the extracts of tobacco leaves. The results were as follows:(1) the amplified chlorophyllase gene AtCLH1 constructed the expression vector p ET28 a-At CLH1 successfully, obtaining the recombinant engineering strain;(2) induced under 30 ℃ for 22 h, the strain could well express the recombinant protein At CLH1 with 0.5 mmol/L IPTG, and the molecular weight was about 35 k Da;(3) the strain showed good chlorophyllase producing capability, and the activity of the produced chlorophyllase could reach up to 24.9 U/m L, which could degrade the chlorophyll in tobacco extract and had a good application prospect in improving the quality of low quality tobacco;(4) based on the results of orthogonal test, the enzyme extract from the strain was added to the tobacco leaf surface, which could make the degradation rate of chlorophyll in the tobacco leaf reach17.06% under the temperature of 37 ℃ at the humidity of 75% for 48 h;(5) after treated by the enzyme liquid, the test tobacco showed increase in the content of aromatic substances, enhancement of tobacco fragrance quality and amount, significant decrease of offensive odor and irritation, significant improvement of agreeable aftertaste, making the overall sensory quality of the tobacco leaf significantly improved.     

基于MATLAB研究油菜叶片的生长动态(英文)

[Ojective]The study aimed to simulate the growth dynamics of rapeseed leaves.[Method]Using nine rapeseed cultivars as experimental materials,the accumulation situation of rapeseed leaves was recorded and their growth dynamics were simulated by using MATLAB program.[Result]The leaf emergence rates of each genotype materials at different growth stages were determined to be:after spring>seedling stage>overwintering stage.All the related indices of the Logistic equation were higher than 0.976 7,commendably reflecting the growth dynamics of rapeseed leaves.[Conclusion]The study provided theoretical basis for the intelligentized cultivation and management and showed the practical guiding significance for the production of high quality rapeseed.     

Expression of HSP72 in Mouse Preimplantation Embryos with Heat Shock

The objective of the paper was to detect HSP72 expression and HSP72 gene sequence in heat shocked mouse preimplantation embryos and the effects of different thermo conditions on hatching rates of embryos. The mouse blastocysts cultured in vitro were heat treated at 40℃ and 38℃ for 1 h, 2 h and 3 h and then recovered at 370C for 3 h, 2 h and 1 h, respectively, to detect their HSP72 gene expression by using RT-PCR after the total R.NA extraction. The hatching rate of the blastocysts for different treated groups was recorded and the expression of liSP72 in the blastocysts was determined by Western blot. The results showed that all the groups of blastocysts, including the control, had the expression of HSP72 gene. The expression of HSP72 protein had the highest level in the embryos stressed at 38℃ for 2 h, and it was significantly higher than that in the control group. The expression of HSP72 in the groups of blastocysts treated at 40℃ was not significantly different from that in the control group. The embryos with induction of mild heat shock at 38℃ for 2 h, then subjected to heat shock at 40℃ for 2 h, had a significant higher （P〈0.05） hatching rate of 54.74% compared to 47.85% in the embryos treated directly at 40℃ for 2 h. The above results indicated that the mouse blastocysts were sensitive to heat shock and a mild heat shock induced HSP72 gene expression. Induction of HSP72 expression with mild heat shock helped embryos to tolerate more severe heat shocks.     

小球藻的分离及其DNA提取方法的研究(英文)

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.     

Isolation of Chlorella vulgaris and Its DNA Extraction Methods

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.     

小球藻的分离及其DNA提取方法的研究(英文)

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.     

A Method Suitable for Extracting Genomic DNA from Animal and Plant——Modified CTAB Method

[Objective] The study aimed to introduce a rapid and effective method that is suitable for extracting genomic DNA from animal and plant. [Method] The genomic DNAs were extracted from tender leaves of 24 peanut cultivars and from the liver,lung and kidney of white mouse through the specifically modified CTAB method. The DNAs were run on agarose gel,next detected by DNA/Protein analyzer. Finally PCR amplification was conducted to detect the quality of DNAs extracted using the modified CTAB method. [Result] The clear and orderly bands were observed in gel detection,and the values of OD260/OD280 for DNAs extracted via modified CTAB method were between 1.77-1.83. The DNAs performed well in PCR amplification. [Conclusion] The DNAs extracted by modified CTAB method could satisfy the requirement of PCR amplification.     

一种适用于动物与植物总DNA提取的方法——改良CTAB法(英文)

[Objective] The study aimed to introduce a rapid and effective method that is suitable for extracting genomic DNA from animal and plant. [Method] The genomic DNAs were extracted from tender leaves of 24 peanut cultivars and from the liver,lung and kidney of white mouse through the specifically modified CTAB method. The DNAs were run on agarose gel,next detected by DNA/Protein analyzer. Finally PCR amplification was conducted to detect the quality of DNAs extracted using the modified CTAB method. [Result] The clear and orderly bands were observed in gel detection,and the values of OD260/OD280 for DNAs extracted via modified CTAB method were between 1.77-1.83. The DNAs performed well in PCR amplification. [Conclusion] The DNAs extracted by modified CTAB method could satisfy the requirement of PCR amplification.     

西川红景天nrDNA ITS序列初步研究(英文)

[Objective] The study aimed to analyze the ITS sequences of nrDNA from Rhodiola alisa and investigate the difference of evolution rate between nrDNA and trnS-trnG and rpl20-rps12 sequences of cpDNA(chloroplast DNA).[Method]Total DNA was extracted from silica-dried leaves of R.alsia by using modified CTAB method.With the extracted DNA sample as template,nrDNA ITS region was amplified,then purified and sequenced.In addition,the yielded ITS sequences were also compared with the known trnS-trnG and rpl20-rps12 sequences of cpDNA from R.alsia.[Result]The ITS sequence of nrDNA from R.alsia was 701 bp in length,of which 13 variable sites were found with a percentage of 1.85%.Of the 13 variable sites,8 were caused by point mutations,5 were the results of insertions or deletions.The(A+T)content and(G+C)content were 46.9% and 53.1%,respectively.The nucleotide diversity(π)was 0.004 27.[Conclusion]The ITS region of nrDNA from R.alsia was more conservative and evolved more slowly than the trnS-trnG and rpl20-rps12 sequences of its cpDNA.     

低温胁迫对甘薯s-腺苷甲硫氨酸合成酶mRNA表达水平的影响

[目的]为研究s-腺苷甲硫氨酸合成酶基因在冷胁迫条件下的表达情况。[方法]分别提取4℃胁迫处理0、12、24、48和72 h后的甘薯叶、茎和块根组织总RNA,通过反转录和实时定量PCR技术分析了该基因的mRNA表达情况。[结果]叶、茎和块根组织中该基因表达量都明显升高,分别于胁迫后24、72和72 h达到表达高峰,其中块根组织表达量增加量最大。[结论]低温胁迫能够促进相关基因的mRNA表达水平升高。     

低温胁迫对甘薯S-腺苷甲硫氨酸合成酶mRNA表达水平的影响

S-腺苷甲硫氨酸合成酶（s-adenosylmethionine synthetase SAMS）是植物代谢中的一个关键酶,它催化甲硫氨酸与ATP生物合成S-腺苷甲硫氨酸（SAM）。SAM是生物合成乙烯以及多胺的前体,参与了植物的转甲基、转氨丙基、转硫反应等多种重要的生理过程。Frank等研究发现,SAM可以与RNA结合参与基因表达调控。近年来研究表明,乙烯和多胺都参与了植物抗逆反应,     

斑马鱼不同发育时期血管内皮生长因子受体2基因表达的实时定量PCR分析

[目的]探讨血管内皮生长因子受体2（VEGFR2-）基因在斑马鱼不同发育时期的表达。[方法]分别从12、24、48、72和96 h的斑马鱼胚胎和仔鱼中提取总RNA,用实时定量RT-PCR方法检测VEGFR2-基因表达,采用2^-△△Ct法进行数据分析。[结果]VEGFR2-基因表达量在12-72 h呈上升趋势,在96 h有所下降。12 h表达量最低,72 h表达量最高,与其他发育期均有显著差异。[结论]斑马鱼血管发育至72 h达成熟阶段。血管发育成熟前,VEGFR2-基因表达水平逐步增长;血管发育成熟后,VEGFR2-基因表达水平下降。     

不同干筋温度对翠碧1号烟叶烘烤质量的影响

[目的]研究不同干筋温度对烟叶烤后质量的影响,为确定不同部位烟叶最佳干筋温度提供理论依据。[方法]以翠碧1号品种为材料,通过设置4个不同干筋温度处理,研究不同处理对下、中、上部位烟叶外观及内在品质的影响。[结果]干筋阶段最高干球温湿度对烤后烟叶的外观质量和内在品质有较大影响,中下部烟叶干筋期最高温度64℃,上部叶68℃,烤后烟叶颜色光泽鲜亮,物理特性好,烟叶的外观等级质量较好,化学成分协调性和感官评吸质量改善效果较为突出。[结论]适当降低烟叶干筋期最高温度对提高烤后烟叶工业可用性较为有利。     

淡竹叶不同部位总黄酮含量的测定

[目的]比较淡竹叶不同部位的总黄酮含量。[方法]用75%的乙醇回流浸提2 h,用NaNO2-A l（NO3）3作显色剂,用紫外-可见分光光度计测吸光度,根据回归方程求出黄酮含量。[结果]淡竹叶不同部位总黄酮含量为：茎叶1.13%、须根0.51%、花序0.36%、块根0.21%。[结论]淡竹叶总黄酮主要集中在茎叶部位,占植株总黄酮量51.13%。     

斑马鱼不同发育时期血管内皮生长因子受体2（VEGFR-2）基因表达的实时定量PCR分析（摘要）

[目的]探讨血管内皮生长因子受体2（VEGFR-2）基因在斑马鱼不同发育时期的表达。[方法]采用Trizol法分别提取12、24、48、72和96h斑马鱼胚胎和仔鱼总RNA;再使用MMLV反转录酶将其反转录为cDNA;合成的cDNA用实时定量RT-PCR方法检测VEGFR-2基因的表达。实时定量RT-PCR扩增反应结束后,利用熔解曲线对扩增产物进行特异性分析。采用2^-△△Ct法进行数据分析。[结果]VEGFR-2基因表达量在12～72h呈上升趋势,在96h时有所下降。12h相对表达量最低,与其他发育期差异极显著（P〈0.01）,72h相对表达量最高,与12、24和96h差异极显著（P〈0.01）,与48h差异显著（P〈0.05）。[结论]斑马鱼血管发育至72h达成熟阶段,血管发育成熟前,VEGFR-2基因表达水平逐步增长;血管发育成熟后,VEGFR-2基因表达水平下降。     

种子低温处理对低夜温下番茄幼苗生长的影响

[目的]研究番茄(Lycopersicon esculentum)种子低温处理对低夜温条件下番茄幼苗生长的影响,探讨番茄幼苗在低夜温下的抗冷性,为低温下获得高质量的番茄幼苗提供理论依据。[方法]以草炭与珍珠岩(2∶1)为育苗基质,番茄L402种子为供试材料。先将番茄L402种子浸种6 h,然后分别于0、4、8和16℃低温条件下进行6、12和24 h低温处理,对照为0 h处理。之后,于16℃下静置0.5 h,再后在25～30℃下催芽,播种于72穴育苗盘中。育苗温度为18～23℃(白天)/6～12℃(夜间)。培养至3片真叶时测定幼苗的各项生长指标(干重、鲜重、根冠比)和生理指标[根系活力、超氧化物歧化酶(SOD)活性及叶绿素、脯氨酸、可溶性蛋白、可溶性糖和丙二醛(MDA)含量]。[结果]种子低温处理能促进低夜温下番茄幼苗的生长,提高幼苗叶绿素含量、可溶性蛋白和脯氨酸的含量,并能增加番茄幼苗在低夜温下的根系活力和SOD活性,同时降低了MDA含量,其中0℃下处理6 h对番茄幼苗在低夜温下的抗冷性提高最大。[结论]种子低温处理能增加番茄幼苗对低夜温的抵抗能力,有利于提高低温季节番茄幼苗的质量。     

低温对烟草叶片中蔗糖向顶端分生组织转运的影响

 【目的】探讨低温胁迫下蔗糖对烟草成花诱导的影响,研究低温条件下烟株中的蔗糖在不同组织中的积累、转运及转运流向。【方法】4℃处理栽培烟草K326 24 h后,检测不同部位组织的蔗糖含量、糖代谢关键酶及其基因表达水平;蔗糖转运蛋白（SUT1）基因表达水平;H2O2含量和抗坏血酸库氧化还原趋势。【结果】与正常培养的对照材料相比,低温处理后蔗糖含量在叶、叶柄、茎和顶端分生组织中呈梯度增大;叶中的蔗糖磷酸合成酶（SPS）活性升高,转化酶（Ivr）活性没有显著变化;淀粉酶活性被明显抑制,淀粉含量增加;叶片中SPSC基因、侧脉中SUT1基因表达水平上调,同时叶中H2O2含量升高,侧脉、叶柄、茎和顶端分生组织中抗坏血酸库趋于氧化态。【结论】低温诱导细胞内环境趋于氧化态势,提高了SUT1的装卸载能力,促进了蔗糖从叶片向顶端分生组织的转运。     

不同采烤方式对红花大金元上部烟叶质量的影响

[目的]探索提高红花大金元上部叶烘烤质量的采烤方式。[方法]研究对比了上部叶(46片)逐片一次性采收、带茎一次性采收、分2次采收(每次采收23片)3种模式,从烤后烟叶外观质量、内在化学成分协调性、经济性状及感官评吸质量等方面进行统计分析。[结果]与上部叶分2次采收模式相比,逐片一次性采收和带茎采收模式,叶片单叶重有所降低,烤后烟叶外观质量、内在化学成分协调性、经济性状及感官评吸质量都有所提高,尤其是对改善第46叶位烟叶整体质量效果明显。综合评价,上部叶逐片一次性采烤表现最好,其次为带茎采烤,分2次采烤表现较差且存在烘烤难度大、杂色比例高、香气质含量低等问题。[结论]在保山植烟区红花大金元上部叶可采取"逐片一次性采烤"或"带茎一次性采烤"模式。     

H2O2对葡萄离体叶片白藜芦醇的诱导作用

[目的]葡萄叶片白藜芦醇含量较虎杖根含量低,难以作为药用提取用途,研究采用诱导剂升高葡萄离体叶片中白藜芦醇的含量.[方法]实验采用H2O2对离体红地球葡萄叶片进行诱导,采用薄层及高效液相色谱法分析白藜芦醇的含量变化,并使用RT - PCR方法分析茋合酶基因(STS)的表达量变化.[结果]H2O2对离体叶片茋类物质具有显著的诱导作用,诱导白藜芦醇的积累呈现显著的量时依赖性;5;H2O2处理48 h后叶片白藜芦醇含量可提高15倍以上,鲜重达到263 μg/g.[结论]H2O2可以作为葡萄离体叶片白藜芦醇的有效诱导剂.