【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.
【學(xué)位授予單位】：東北林業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2018
【分類號】：S792.11;S714.3
【圖文】：

２．４結(jié)果與分析逡逑將構(gòu)建好的植物表達(dá)載體ｐＣ－ｐＰｒｏ－２－ＴＣＳ經(jīng)凍融法轉(zhuǎn)化到根癌農(nóng)桿菌ＥＨＡ１０５，結(jié)逡逑果如圖２－２所示，每一個加樣槽顯示樣品均擴(kuò)增出一條分子量約為７５０ｂｐ的特異條帶。逡逑結(jié)果表明，含有ＴＣＳ啟動子及結(jié)構(gòu)基因的ｐＣ－ｐＰｒｏ－２－ＴＣＳ植物表達(dá)載體己成功轉(zhuǎn)入了根逡逑癌農(nóng)桿菌ＥＨＡ１０５中，得到用于轉(zhuǎn)化的工程菌。邐逡逑：\，｜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＋：陽性對照；ＣＫ—：陰性對照；逡逑１，２，３：質(zhì)粒邋ｐＣ－ｐＰｒｏ－２－ＴＣＳ：逡逑圖２－２農(nóng)桿菌質(zhì)粒ＰＣＲ檢測電泳分析圖譜逡逑Ｆｉｇ．２－２邋Ｐｌａｓｍｉｄ邋ＰＣＲ邋ｆｏｒ邋ｅｌｅｃｔｒｏｐｈｏｒｅｓｉｓ邋ａｎａｌｙｓｉｓ逡逑２．５本章小結(jié)逡逑采用氯化鈣法成功制備了農(nóng)桿菌感受態(tài)，成功的獲得了含有ＴＣＳ啟動子及結(jié)構(gòu)基逡逑因的ｐＣ－ｐＰｒｏ－２－ＴＣＳ載體用于轉(zhuǎn)化的工程菌菌株。逡逑－１０－逡逑

３．４結(jié)果與分析逡逑３．４．１山新楊組培苗的擴(kuò)繁逡逑約２０ｄ以后，可見到接入的山新楊外植體得到很好的擴(kuò)繁效果，如圖３－１。得到逡逑４５０瓶無污染、叢生芽長而茂密、生長良好的山新楊組培苗。逡逑邐瞧逡逑圖３－１邋２０ｄ培養(yǎng)后的山新楊組培苗逡逑Ｆｉｇ．３－１邋Ｔｈｅ邋ｓａｐｌｉｎｇｓ邋ｏｆ邋Ｐｏｐｕｌｎｓ邋ｄａｖｉｄｉａｎａ＇ｘＲｂｏｌｌｅａｎａ邋ａｆｔｅｒ邋２０ｄ逡逑３．４．２抗性芽的篩選逡逑利用根癌農(nóng)桿菌轉(zhuǎn)化體系，最后獲得６株具有潮霉素抗性芽。如圖３－２逡逑｜｜．．．邋Ｍ邋■ｊＬ逡逑．一歌．入．．．邐．逡逑圖３－２篩選出的在生根培養(yǎng)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

３．４結(jié)果與分析逡逑３．４．１山新楊組培苗的擴(kuò)繁逡逑約２０ｄ以后，可見到接入的山新楊外植體得到很好的擴(kuò)繁效果，如圖３－１。得到逡逑４５０瓶無污染、叢生芽長而茂密、生長良好的山新楊組培苗。逡逑邐瞧逡逑圖３－１邋２０ｄ培養(yǎng)后的山新楊組培苗逡逑Ｆｉｇ．３－１邋Ｔｈｅ邋ｓａｐｌｉｎｇｓ邋ｏｆ邋Ｐｏｐｕｌｎｓ邋ｄａｖｉｄｉａｎａ＇ｘＲｂｏｌｌｅａｎａ邋ａｆｔｅｒ邋２０ｄ逡逑３．４．２抗性芽的篩選逡逑利用根癌農(nóng)桿菌轉(zhuǎn)化體系，最后獲得６株具有潮霉素抗性芽。如圖３－２逡逑｜｜．．．邋Ｍ邋■ｊＬ逡逑．一歌．入．．．邐．逡逑圖３－２篩選出的在生根培養(yǎng)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

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