转TCS基因山新杨的种植对土壤微生物生态系统的影响

发布时间：2020-07-14 04:45

【摘要】：The species of Populus spp.are important trees planted worldwide,and play crucial roles in forest industry in our country.At present,the application of transgenic biotechnology in poplar genetic breeding has accelerated the selection process of its fine new varieties.However,the problem of transgenic ecological safety caused by exogenous gene has become the focus of current research.Therefore,it is of great significance to evaluate the ecological safety of transgenic trees in theory research and production application.This study to the TCS gene Populus davidiana×P.bolleana of radix trichosanthis protein fusion protein gene and to study its genetic traits after change of rhizosphere secretion influence on soil microorganisms and soil physical and chemical properties,in order to evaluate the release of transgenic plants and its gene product may cause risk of soil ecological environment to provide technical information.The main results are as follows:(1)This study has found that TCS gene Populus davidiana×P.bolleana have significant effects on five kinds of soil enzyme activities(P0.05),indicating that the change of soil enzyme activity was related to the new root exudate of TCS gene.(2)This study has found that the TCS gene Populus davidiana×Pbolleana plant after 30d-210d period compared with the control soil,the soil turned Populus davidiana×P.bolleana root secretion of TCS gene mountain index with different degree of impact of soil physical and chemical properties,in addition to the content of soil organic matter in part time difference was significant(P0.05).This indicates that the soil physicochemical property index may be affected by the transgenic TCS gene mountain new poplar secretions,which has a great influence on some time periods.(3)In this study,five kinds of soil enzyme activities and 6 soil physical and chemical properties indexes were analyzed,and most of the indexes were related to each other,and some of them reached significant or significant levels.This indicates that there is a close relationship between five soil enzyme activities and six physicochemical properties indexes in the study.(4)Soil microbial quantity of research results show that TCS gene Populus davidiana×P.bolleana root soil and contrast the three groups all show the number of bacteria is higher than actinomyces and fungi,the quantity of fungi are reached the highest at 90d,and significant difference(P0.05).And new genetically modified(gm)mountain Yang compared with controlled soil and the three major groups,the number of fungi and actinomycetes were less than non-gm new Yang mountain soil,in the number of fungi in 90d reached significant difference(P0.05),number of actinomycetes have reached significant difference except 210d(P0.05);The number of bacteria in 30d-210d was higher than that in the latter group and the difference was significant(P0.05).This indicates that the soil can be cultivated by the three groups of microorganism,which may be affected by the transfer of TCS gene Populus davidianaxP.bolleana secretions.(5)This study using the method of Biolog Eco micro tablet on soil microbial community functional index calculation and analysis,the results show that the new turn TCS gene Populus davidianaxP.bolleana soil microbial AWCD value trend compared with controls,change in the 150d-210d and AWCD value higher than the control;The diversity index of Shannon-Winner,Simpson and McIntosh was all lower than the control group except 30d,and the difference was significant.It is indicated that the secretion of Populus davidianaxP.bolleana root exudate of TCS gene has a certain inhibitory effect on soil microbial diversity.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2018
【分类号】：S792.11;S714.3
【图文】：

农杆菌,电泳分析,质粒,图谱

２．４结果与分析逡逑将构建好的植物表达载体ｐＣ－ｐＰｒｏ－２－ＴＣＳ经冻融法转化到根癌农杆菌ＥＨＡ１０５，结逡逑果如图２－２所示，每一个加样槽显示样品均扩增出一条分子量约为７５０ｂｐ的特异条带。逡逑结果表明，含有ＴＣＳ启动子及结构基因的ｐＣ－ｐＰｒｏ－２－ＴＣＳ植物表达载体己成功转入了根逡逑癌农杆菌ＥＨＡ１０５中，得到用于转化的工程菌。逦逡逑：\，｜逡逑一■邋Ｖ逡逑－０００－ＪＰ邋－邋＇逡逑ｉ00ｂｐ邋－．．１逡逑Ｍ：邋Ｍａｋｅｒ邋ＤＬ２０００：邋ＣＫ＋：阳性对照；ＣＫ—：阴性对照；逡逑１，２，３：质粒邋ｐＣ－ｐＰｒｏ－２－ＴＣＳ：逡逑图２－２农杆菌质粒ＰＣＲ检测电泳分析图谱逡逑Ｆｉｇ．２－２邋Ｐｌａｓｍｉｄ邋ＰＣＲ邋ｆｏｒ邋ｅｌｅｃｔｒｏｐｈｏｒｅｓｉｓ邋ａｎａｌｙｓｉｓ逡逑２．５本章小结逡逑采用氯化钙法成功制备了农杆菌感受态，成功的获得了含有ＴＣＳ启动子及结构基逡逑因的ｐＣ－ｐＰｒｏ－２－ＴＣＳ载体用于转化的工程菌菌株。逡逑－１０－逡逑

潮霉素抗性,山新杨,生根培养基,植株

３．４结果与分析逡逑３．４．１山新杨组培苗的扩繁逡逑约２０ｄ以后，可见到接入的山新杨外植体得到很好的扩繁效果，如图３－１。得到逡逑４５０瓶无污染、丛生芽长而茂密、生长良好的山新杨组培苗。逡逑逦瞧逡逑图３－１邋２０ｄ培养后的山新杨组培苗逡逑Ｆｉｇ．３－１邋Ｔｈｅ邋ｓａｐｌｉｎｇｓ邋ｏｆ邋Ｐｏｐｕｌｎｓ邋ｄａｖｉｄｉａｎａ＇ｘＲｂｏｌｌｅａｎａ邋ａｆｔｅｒ邋２０ｄ逡逑３．４．２抗性芽的筛选逡逑利用根癌农杆菌转化体系，最后获得６株具有潮霉素抗性芽。如图３－２逡逑｜｜．．．邋Ｍ邋■ｊＬ逡逑．一歌．入．．．逦．逡逑图３－２筛选出的在生根培养基中的潮霉素抗性植株逡逑Ｆｉｇ．３－２邋Ｓｃｒｅｅｎｉｎｇ邋ｏｆ邋ｈｙｇｒｏｍｙｃｉｎ邋ｒｅｓｉｓｔａｎｃｅ邋ｐｌａｎｔｓ

山新杨,组培苗

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