The design of aided software for osmotic stress responding genes mining in plant genome

A software and algorithm which based on random sequence model uses osmotic stress responding cis elements from existing information sources of biology was designed.It can infer the genic downstream function of Arabidopsis thaliana through analyzing its promoter region,and can offer effective aided analysis to mine osmotic stress responding genes in Arabidopsis thaliana genome.The practical application proves that this software can aid to analyze vast genic data and offer important data evidence.     

用最短路径传递共表达预测拟南芥基因功能

提出基因之间传递共表达可作为一个重要因素来连接同一代谢通路中的基因,而同一代谢通路中的功能相似的基因都是高表达相关的。因此可通过求代谢通路下的最短路径,由同在一条最短路径上的与未知基因高表达相关的已知基因来预测未知基因的功能。通过用最短路径算法分析拟南芥代谢通路下的共表达数据对未知基因的功能进行预测,证明了此方法可以预测出拟南芥代谢通路下未知基因的功能,并验证了通过在代谢通路下求最短路径来预测基因功能的方法具有一定的可行性和有效性。     

Cloning and Expression Level Analysis of Two BnaANT Candidate Genes in Brassica napus

AINTEGUMENTA （ANT） gene has been proved to have a novel function on controlling the organ size and the seed weight in Arabidopsis thaliana, the research on its orthologous gene in Brassica napus will be of potential interest to elucidate the molecular mechanism of rapeseed development and increase the rapeseed output. A SMART cDNA library of the flower bud from B. napus cultivar Zhongshuang 9 was constructed, and the full-length cDNAs of two homologous BnaANT candidate genes were cloned by PCR in B. napus, namely BnaX.ANT.a （GenBank accession no. DQ211969） and BnaX.ANT.b （accession no. DQ211970）. They shared 87.1 and 83.4% amino acid identity with ANT of A. thaliana, respectively. There is 87.4% amino acid identity between BnaX.ANT.a and BnaX.ANT.b. By quantitative real-time PCR, the obviously differential expression levels of the two BnaANT candidate genes were detected in the 28 DAF seeds of the big and small grains B. napus lines, the expression abundance of BnaANT in the 28 DAF seed of 9311 was over three times compared to that of 9260, which indicates that these two BnaANT genes are also likely related to the floral organ size and the seed weight in B. napus.     

雄性育性基因RNA干扰载体的构建与鉴定(英文)

cDNA fragment of fertility gene MS2 from cotton was cloned by RT-PCR approach, it was highly homologous with relevant genes of Brassica napus and Arabidopsis thaliana. According to the principles of constructing RNAi vector, sense and antisense fragments of MS2 gene carrying restriction endonuclease recognition sites were amplified via PCR technique, ligated with the first intron of upland cotton chinase gene, then inserted into artificially modified plant expression vector pBI121, yielding RNAi vector pBGP12MSIn. The results showed that RNAi vector pBGP12MSIn harboring MS2 gene driven by anther specific promoter BGP was successfully constructed. Our results laid a foundation for studying the function of this gene and genetic transformation of plant male sterile lines.     

Isolation and Characterization of a Cinnamoyl CoA Reductase Gene（CCR） in Pear（Pyrus pyrifolia）

Pear is a popular and commercially important fresh fruit, and its texture is related to the presence of sclereid formatted by parenchyma cell with lignification in vascular plants. Previous studies have demonstrated that content of lignin may be regulated by cinnamoyl CoA reductase（CCR） in various plants. However, the function of CCR in pears remains very limited. In the present study, we isolated a cDNA encoding CCR（PpCCR, GenBank accession No. KF999958） and its promoter（proPpCCR） from Whangkeumbae pear to investigate the function of CCR in lignin biosynthesis. PpCCR-GFP expressed in rice mesophyll protoplast demonstrated that PpCCR-GFP was localized in the cytoplasm, indicating that CCR may function in cytoplasm without localization signals. In transgenic plants carrying PpCCR, we observed higher lignin content compared with that in wild type plants, further suggesting that PpCCR can affect the lignin contents through regulating lignin biosynthesis in Arabidopsis thaliana. More studies in other plants are needed to confirm our conclusion.     

牛体外发育胚胎特定阶段差异表达基因的研究

[目的]研究不同发育时期牛体外受精胚胎在基因表达模式上的差异.[方法]利用单个胚胎mRNA差异显示技术,对单个8细胞期胚胎与囊胚进行mRNA差异显示,获得1条特异表达条带,对其进行克隆、测序,并与GenBank进行对比.[结果]该序列与牛核糖体蛋白131基因(ribosomal protein 131,RPL3])具有99%的同源性.采用实时定量PCR技术检测8细胞期和囊胚期胚胎RPL31的mRNA表达量,结果表明,RPL31在8细胞期胚胎的相对表达量为囊胚期胚胎的3.2倍.[结论]为揭示和阐明控制牛早期胚胎发育的相关机理提供依据. Abstract： The cattle different stage embryos obtained from in vitro was studied using the technology of single preimplantation embryo mRNA different display:single 8-cell and blastocyst stage embryos were studied using technology of mRNA different display and one different fragment was found.The result suggested that this fragment displayed high homology (99%) to cattle mRNA for ribosomal protein L31.Then to detect the expression of RPL31 mRNA in 8 cell and blastocyst stage embryos by real-time quantitative PCR,the result showed the relative amount of 8 cells was 3.2 times of blastocyst's.     

质粒pUC19-CM-D的构建及应用

[目的]用克降载体pUC19构建乳杆菌自杀质粒及乳杆菌基因缺失工程菌.[方法]以鼠李糖乳杆菌为研究对象,在质粒pUC19的多克隆位点插入氯霉素抗性基因构建pUC19-CCM载体;在pUC19-CM载体氯霉抗性基因的两侧均添加1个用于同源重组的同源臂,再构建成自杀质粒pUC19-CM-D.将自杀质粒pUC19-CM-D.将自杀质粒pUC19-CM-D转化乳杆菌进行抗生筛选即可得到目标基因被抗性基因替换的突变株.[结果]通过改造pUC19质粒,获得了可应用于构建鼠李糖乳杆菌D-乳酸脱氢酶基因缺陷型菌株的自杀质粒pUC19-CM-D.运用抗性基因替换方法来构建基因缺陷型菌株,在一定程度上避免了质粒整合带来的问题,同时提供了一个筛选标记,能够通过一次筛选即可得到缺陷型菌株.这种自杀质粒的改造和基因缺陷型菌株的构建方法具有通用性,可以应用于其他的载体和菌株.[结论]pUC19-CM-D质粒的构建及应用为乳杆菌基因缺失工程菌的构建提供了1个快速有效的手段,也为乳杆菌基因功能研究奠定了基础. Abstract： [Objective] The aims were to construct a new suicide plasmid of Lactobacillus and gene deletion engineering bacteria of Lactobacillus with pUC19 vector,[Methods]pUC19-CM was constructed by inserting a chloramphenicol resistant gene into the multi-cloning site of pUC19,and then two homologous fragments were cloned into each side of the pUC19-CM to construct suicide plasmid pUC19-CM-D.[Results]A replacement mutant strain,whose target gene was replaced by resistant gene,could be obtained by transforming the suicide plasmid pUC19-CMD into Lactobacillus for resistance screening. [Conclusion] The construction and application of pUC19-CM-D provided a fast and efficient means of construction of gene deletion engineering bacteria of Lactobacillus,and laid a foundation for study of gene function of Lactobacillus.     

基于蒙特卡洛多重积分的DNA动力学结构参数的计算

依据最新NDB数据库中蛋白质-DNA复合物晶体结构数据,基于修正的DNA结构统计力学模型,利用蒙特卡洛多重积分计算DNA动力学结构的有关参数,并对计算得到的结果进行时间复杂度和精确度分析. Abstract： Based on protein-DNA complex crystal structural data in up-to-date Nucleic Acid Database,the related parameters of DNA Kinetic Structure were investigated by Monte-Carlo Multiple Integrals on the base of modified DNA structure statistical mechanical model,and time complexity and precision were analyzed on the calculated results.     

抗黄瓜花叶病毒RNAi载体的构建及烟草的转化

[目的]构建抗黄瓜花叶病毒RNAi载体,并将载体转入烟草.[方法]采用RT-PCR方法,扩增黄瓜花叶病毒NS04加工番茄分离物的RNA2基因组的序列选取CMV RAN2基因组中的复制酶片段作为靶序列,构建pBi35SCR2真核表达载体,并对表达载体时行鉴定;通过农杆菌介导的方法将表达载体转入烟草,用PCR的方法检测载体是否转入.[结果]系统进化树分析结果表明,RNA2中编码CMV-2a的序列与中国浙江的DQ412731 分离物有较高核苷本乡酸及氨基酸同源性,分别达到98.0%和96.5%;RCR结果表明,试验成功构建了pBi35SCR2真核表达载体,并成功将表达载体转入烟草[结论]试验获得的转基因烟草可作为后期攻毒试验的材料,并为研究加工番茄抗黄瓜花叶病毒奠定了基础. Abstract： [Objective] Aimed to construct RNAi vector resistant to cucumber mosaic virus and transferred this vector into tobacco.[Method]RT-PCR method was used to amplify cucumber mosaic virus NSO4 and process RNA2 gene sequen of tomato isolates.The analysis results of phylogenetic tree demonstrated that the sequence in RNA2 encoded CMV-2a had 98.0% and 96.5% homology with nucleotide and amino acid of DQ412731 isolate of Zhejiang,China.The replicase fragment in CMV RAN2 gene was taken as target sequence to construct pBi35SCR2 eukaryotic expression vector,then the expression vector was identified.Through agrobacterium-mediated method,the expression vector was transferred into tabacco and PCR method was used to check the transfer.The PCR results demonstrated that the experiment had successfully construct eukaryotic expression vector of pBi35SCR2 and the expression vector was successfully transferred into tabacco. [Conclusion] The obtained transgenic tobacco could be used as challenge test material in following experiment and provided foundation for studying processing tomato resist cucumber mosaic virus.     

底栖鱼类对水田上覆水中磷素动态的扰动效应

[目的]研究底栖鱼类泥鳅对水田上覆水中磷素动态的扰动效应,探讨生物扰动机制.[方法]基于模拟试验,使用离子色谱法和分光光度法,对比分析上覆水中磷素含量在有/无泥鳅活动时的差异.[结果]扰动组的TP、DTP和PP浓度在试验开始阶段与对照组无显著差异,在试验中、后期显著高于对照(P<0.05).扰动组要的PP/TP高于对照组,扰动组中TP浓度的增加主要是由于PP的增加,扰动组的DIP/DT在试验中、后期显著高于对照(P<0.05).[结论]底栖鱼类对水田上覆水中的磷素产生了扰动作用,增加了水稻生长可利用的的磷素养分. Abstract： [Objective] The research aimed to investigate the bioturbation effects of benthic fish Misgurnus anguillicaudatus on phosphorus dynamic in overlying water of paddy field,as well as to explore the bioturbation mechanism.[Method]Based on simulation experiment,the phosphorus contents in overlying water were analyzed comparatively with and without Misgurnus anguillicaudatus by the using of ion chromatography and spectrophotometry. [Result] The concentrations of total phosphorus (TP),dissolved total phosphorus(DTP)and particular phosphorus(PP) in bioturbation group had no significant differences with those in control group in initial stage of experiment,and became significantly higher than control group in middle and late stages of experiment(P<0.05).The PP/TP ratios in bioturbation group were bigger than those in control group,the increase of TP concentration in bioturbation group was mainly due to the increase of PP.The ratios of dissolved inorganic phosphorus(DIP) to DTP (DIP/DTP) were significantly bigger than those in control group in middle and late stages of experiment (P<0.05). [Conclusion] The benthic fish had bioturbation effects on phosphorus in overlying water of paddy field,which increased the available phosphorus for rice growth.     

一株产紫杉醇的曼地亚红豆杉内生真菌的分离及鉴定

[目的]分离并鉴定1株产紫杉醇的曼地亚红豆杉内的内生真菌.[方法]从曼地亚红豆杉树皮内表皮中分离得到32株内生真菌,并通过高效液相色谱法检测其发酵产物.[结果]筛选获得1株可以产紫杉醇的内生真菌M57,其紫杉醇产量为45～50μg/L,并通过对M57菌落的形态学观察以及18S rDNA序列分析初步将其鉴定为根霉属(Rhizopus)真菌.[结论]该菌株的发现为微生物发酵法生产紫杉醇提供了具有潜在应用价值新的菌种. Abstract： [Objective] The aim was to isolate and identify a taxol-producing endophytic fungus from Taxus media.[Method]32 strains of endophytic fungi were identified form the inner bark of T.media,and their fermentation products were detected by high performance liquid chromatography (HPLC). [Result] Through the screening,a strain of taxol-producing endophytic fungi M57 was obtained,which could produce 45-50 μg/L of taxol,and M57 was defined as Rhizopus sp.through morphological observation and 18S rDNA sequence analysis. [Conclusion] The finding of Rhizopus sp.M57 provided a promising strain for producing taxol with taxol-producing fungi fermentation process.     

电场作用下人工湿地植物的生理生化响应

通过测定人工湿地植物叶片中叶绿素含量、MDA含量及SOD酶活性.研究不同通电强度下人工湿地植物生理特性变化,分析电场对植物生理特性的影响机理,为利用电场强化人工湿地去除污染物提供依据.研究结果表明,与对照植物相比,1和3 V低强度电压对植物正常生理指标变化无明显影响,且生长趋势优于对照组;随着电压强度的升高,植物叶片中叶绿素含量、MDA含量以及SOD酶活性受到较大影响,表明植物体受到较强的氧化胁迫,生长受到危害.合适的电场能够强化人工湿地的污水处理效果. Abstract： By measuring wetland plants chlorophyll content,malondialdehyde (MDA) content and superoxide dismutase (SOD) enzyme activity,the changes of wetland plant physiological characteristics under different power strength were studied,and the mechanism of electric field on plant physiological characteristics was analyzed to provide a theoretical basis for the pollutant removal ability strengthening of artificial wetland under electric field.The results showed that compared with the control plants,low-intensity-voltage (1 V and 3 V) had no significant effect on the normal physiological and biochemical indexes of the plants,and the growth trend was better than the control group;with the voltage increasing,plant chlorophyll content,MDA content and SOD activity were greatly affected,indicating that plants were under strong oxidative stress,and the growth was damaged.Therefore,a suitable electric field could enhance the sewage treatment effect of constructed wetland.     

不同土地利用方式下球囊霉素相关土壤蛋白与有机碳及土壤质地的关系

[目的]揭示土地利用方式对球囊霉素相关土壤蛋白(GRSP)的影响.[方法]测定了海南岛5种主要土地利用方式下GRSP、土壤有机碳(Soil oaganic carbon)与土壤质地(Soil texture)在0～10、10～20、20～50cm3个土壤层次中的分布情况.[结果]相对于保持较好的次生林,4种不同耕作土壤的GRSP与有机碳含量均有所下降.GRSP在海南岛5种主要土地利用方式土壤中的含量为0.53～4.80 mg/g,占有机碳的百分比为7.9%～23.4%,是碳库的重要组成部分.GRSP占有机碳的比例在不同土地利用方式下差异显著,在土层之间差异不显著,GRSP和有机碳含量在土壤剖面上垂直分布较明显.GRSP与有机碳、砂粒含量呈显著正相关,与粉粒和粘粒呈显著负相关.[结论]砂粒含量在很大程度上决定了CRSP的含量,壤土相对粘土更有利于GRSP的累积. Abstract： [Objective] The aim was to reveal the effects of different land use types on soil composition.[Method]GPRS,Soil organic carbon content and soil texture in 3 depths(0-10,10-20,20-50 cm)of 5 main kind of selected land use type were examined in Hainan. [Result] The results showed that GRSP and SOC content of four artificial land use types decreased compared with the natural sacondary foreSt land,the GRSP content of all samples ranged from 0.53-4.80 mg/g,accounting for 7.9%-23.4% of the SOC,which means that GRSP was one important component of SOC pool in soil.The ratio of GRSP to SOC was signiflcantly different among land use types but the depths.GRSP and SOC exhibited obvious vertical distribution pattern.GRSP was significantly positively related to SOC and sand content but negatively related to silt and clay content. [Conclusion] The sand content determined the GRSP content significantly and Joarn was better matrix for GRSP accumulation than clay.     

山区生态友好型土地利用对策研究——以重庆市黔江区为例

以黔江区为研究对象,通过对生态环境影响因子的分析,发现制约黔江区土地利用环境发展的因素主要有区域气候变化、土壤要素、水文环境以及水土流失和土地退化等.并任以上分析的基础上,进一步分析了土地利用变化对生态环境的影响和制约黔江生态环境及区域发展的因素,最后提出了适应山区土地利用与生态环境协凋的对策,对黔江社会经济发展和生态建设具有重要意义. Abstract： Accompanying the development of social economy,the land use model of mountainous area,typically eco-weak area,is changing gradually.Here the establishment of eco-friendly land use model in mountainous area,will pioneer the model of sustainable development in that area.Concerning Qianjiang District of Chongqing Municipality,huge change of land use model,mainly embodied in the unceasing increase of construction land and gradual decrease of agricultural use land,has taken place in recent years.To explore the eco-friendly land use model in mountainous area,Qianjiang District was chosen as the study object in the present study.Via analyzing the changes in land use model,we found that related eco-environment restrictive factors mainly regional,climatic change,soil texture,hydrological environment as well as soil erosion and land degradation,etc.And based on these results,we further analyzed the effect of land use change on eco-environment and the factors restricting the maintenance of eco-environment and regional development,finally put forward the counter measures for balancing land use and co-environment in mountainous area.The results will be important for the development of social economy and eco-system construction in Qianjiang District.     

催产素在广西本地水牛下丘脑-垂体-卵巢轴的免疫组化定位

[目的]研究催产素(OT)在广西本当水牛下丘脑、垂体及卵巢的分布上的联系,进而了解OT在下丘脑、垂体及卵巢3者之间分泌释放途径.[方法]采用免疫组织化学SuperPicTureTM二步法检测5头广西本地水牛的下丘脑、垂体和卵巢中催产素(OT)的分布情况.[结果]下丘脑中分泌OT的神经元主要分布在弓状核、视上核及室旁核,在腹内侧核、腹外侧核、交叉上核、背内侧核、乳头体、下丘脑前核等核团也有一定数量的阳性神经元;在腺垂体发现OT免疫反应阳性产物,自垂体柄和正中隆起的一侧可见到平行排列的OT阳性神经纤维断续地延伸至神经部;卵巢中只在生殖上皮、卵泡颗粒细胞和黄体细胞发现有大量OT免疫阳性产物.[结论]首次发现OT存在于广西本地水牛下丘脑-垂体-卵巢轴的各个环节,并且下丘脑中各主要核团均有OT免疫阳性神经元的分布,尤其以弓状核、视上核、室旁核分布最多,为进一步研究OT的合成和作用机制提供形态学依据,并对广西本地水牛的繁殖育种及泌乳起到一定的参考指导作用. Abstract： To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochemistry SuparPicTureTM two step method was used to detect the distribution of OT in hypothalamatic-pituitary-ovary axis of five femal Guangxi local buffalo.The test results could provide morphology according to study the OT's synthesis and mechanism of action ,and could play reference and directions part in breeding Guangxi local buffalo.The test results display:oxytocin immuno reactive (OT-IR) neuronsw eremainly distributed arcuate nucleus,supraoptic nucleus and paraventricular nucleus,and OT-IR neurons was also found in ventrornedial nucleus,ventrolateralis nucleus,suprachiasrnaticus nucleus,dorsomediai nucleus,mamillary body,anterior hypothalamic nucleus and so on.The OT immunoactive production was found in pituitary and few OT-IR nerve fibers extended to post pituitary from hypophyseal stalk and medium eminence.In ovaries,OT immunoactive productions were only distributed in germinal epithelium cells,granulosa cells and lutein ceils.The OT was first discovered in singulorum link of hypothalamatic-pituitary-ovary axis of Guangxi local buffalo.The OT immunoactive neurons were first discovered in every main nucleus of Guangxi local buffalo hypothalamus,especially distributed in arcuate nucleus,supraoptic nucleus and paraventricular nucleus.