本文選題：cryAb + epsps�。� 參考：《農(nóng)業(yè)環(huán)境科學(xué)學(xué)報》2017年10期

【摘要】：為探討轉(zhuǎn)基因玉米的生態(tài)安全問題,2015年分別于玉米拔節(jié)期、抽雄期、乳熟期和完熟期采集土壤樣品,并采用熒光定量PCR(Quantitative real-time PCR,q PCR)和末端限制性片段長度多態(tài)性分析(Terminal restriction fragment length polymorphism,TRFLP)技術(shù),研究了轉(zhuǎn)cry1Ab和epsps基因玉米C0030.3.5種植對土壤古菌豐度和多樣性的影響。結(jié)果表明:轉(zhuǎn)cry1Ab和epsps基因玉米C0030.3.5(TM)和受體玉米DBN318(PM)的根際土和非根際土古菌數(shù)量為1.41×10~9~4.04×10~9copies·g~(-1)土,隨生長時期的推進均呈現(xiàn)先升高后降低的變化趨勢,同一生長時期和土壤樣品采集區(qū)域,2種玉米古菌16S rRNA基因豐度間無顯著差異。T-RFLP分析共獲得15種不同長度的T-RFs,其中89 bp和184 bp片段為優(yōu)勢種群,同一生長時期和土壤樣品采集區(qū)域每種優(yōu)勢種群在TM和PM間均無顯著差異。土壤古菌Shannon指數(shù)除PM非根際土呈現(xiàn)先降低后升高的變化趨勢外,其他均表現(xiàn)為先降低后升高再降低;TM和PM根際土古菌Evenness指數(shù)的變化趨勢均為先降低后升高,而非根際土古菌Evenness指數(shù)表現(xiàn)為先降低后升高再降低。同一生長時期和土壤樣品采集區(qū)域TM和PM的Shannon指數(shù)間及Evenness指數(shù)間差異均不顯著。冗余分析(Redundancy analysis,RDA)顯示土壤總氮和硝態(tài)氮含量對古菌群落結(jié)構(gòu)有顯著影響。主成分分析(Principal component analysis,PCA)表明,TM和PM土壤古菌群落在根際土和非根際土中均未發(fā)生明顯分離,說明TM和PM土壤古菌群落組成無顯著差異。上述結(jié)果表明:轉(zhuǎn)cry1Ab和epsps基因玉米C0030.3.5土壤古菌豐度和群落結(jié)構(gòu)組成與受體玉米DBN318無顯著差異,古菌豐度和多樣性主要受生長時期的影響,受土壤樣品采集區(qū)域的影響不顯著,土壤總氮和硝態(tài)氮是調(diào)控土壤古菌群落變化的關(guān)鍵因子。
[Abstract]:In order to study the ecological safety of transgenic maize, soil samples were collected in jointing, heading, milking and ripening stages in 2015, respectively. The effects of cry1Ab and epsps gene maize (C0030.3.5) on the abundance and diversity of soil paleomycetes were studied by means of fluorescence quantitative PCR(Quantitative real-time PCRQ PCR and terminal restriction fragment length polymorphism (TRFLP) technique. The results showed that the number of archaic bacteria in rhizosphere soil and non-rhizosphere soil of transgenic cry1Ab and epsps gene maize C0030.3.5 and receptor maize DBN318PM) was 1.41 脳 10 ~ (9) ~ (9) 脳 10~9copies ~ (-4) 脳 10~9copies ~ (-1), which increased at first and then decreased with the development of growth period, and the results showed that the number of bacteria in rhizosphere and non-rhizosphere soil was 1.41 脳 10 ~ (9) and 4.04 脳 10~9copies / g ~ (-1), respectively. There was no significant difference in the abundance of 16s rRNA gene between two species of maize palaeomycetes in the same growing period and soil sample collection region. T-RFLP analysis showed that 15 T-RFSs of different lengths were obtained, of which 89bp and 184bp were dominant populations. There was no significant difference between TM and PM in each dominant population in the same growing period and soil sampling area. The Shannon index of soil palaeomycetes decreased first and then increased, except that PM decreased first and then increased, and then decreased and then decreased, and the Evenness index of Palaeobium in PM rhizosphere soil decreased first and then increased. The Evenness index of non-rhizosphere bacteria decreased first and then increased and then decreased. There was no significant difference in Shannon index and Evenness index between TM and PM in the same growing period and soil sample collection area. Redundancy analysis showed that the contents of soil total nitrogen and nitrate nitrogen had a significant effect on the community structure of Palaeomycetes. Principal component analysis (PCA) showed that there was no significant separation of soil paleomycetes community between TM and PM soil in rhizosphere and non-rhizosphere soil, indicating that there was no significant difference between TM and PM soil paleomycetes community composition. The results showed that there was no significant difference between C0030.3.5 soil paleomycetes abundance and community structure composition of transgenic cry1Ab and epsps gene maize and that of recipient maize DBN318. The abundance and diversity of paleomycetes were mainly affected by growth period, but not by soil sample collection area. Soil total nitrogen and nitrate nitrogen are the key factors to regulate the change of soil paleomycetes community.
【作者單位】： 農(nóng)業(yè)部環(huán)境保護科研監(jiān)測所;天津農(nóng)學(xué)院農(nóng)學(xué)與資源環(huán)境學(xué)院;
【基金】：轉(zhuǎn)基因生物新品種培育重大專項(2015ZX08013002-004,2016ZX08012005-005) 國家自然科學(xué)基金項目(31200424)~~
【分類號】：S154.3;S513

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