轉(zhuǎn)EPSPS基因抗除草劑大豆對土壤原核微生物群落生態(tài)影響的研究
發(fā)布時間：2019-05-31 20:38
【摘要】：隨著植物轉(zhuǎn)基因技術(shù)的發(fā)展,該技術(shù)在農(nóng)業(yè)領(lǐng)域中得到廣泛應(yīng)用。1996年,美國的第一例商業(yè)化的抗草甘膦轉(zhuǎn)基因大豆被批準(zhǔn)種植,從此轉(zhuǎn)基因大豆的種植面積也逐漸增加,其中絕大多數(shù)是抗除草劑大豆。轉(zhuǎn)基因大豆給人類帶來了巨大經(jīng)濟(jì)效益和社會效益,但是其對環(huán)境的安全性問題也逐漸引起了人們的關(guān)注,尤其是對土壤生態(tài)環(huán)境的影響。土壤中的微生物對外界干擾反應(yīng)非常靈敏,轉(zhuǎn)基因作物的外源基因及其表達(dá)產(chǎn)物在土壤生態(tài)系統(tǒng)中的累積很可能會對土壤中的微生物群落存在一定程度的影響,而且大豆與土壤中的固氮相關(guān)微生物群落關(guān)系密切,因此在評價其環(huán)境安全性的研究中,對土壤微生物群落多樣性變化的研究,特別是對土壤固氮相關(guān)微生物群落的影響是非常必要的。本研究選取的種植點(diǎn)及大豆品種分別為：內(nèi)蒙古種植點(diǎn)的轉(zhuǎn)EPSPS基因抗除草劑大豆NZL06-698,受體品種是蒙豆12和區(qū)域?qū)φ沾蠖购诤?3；吉林種植點(diǎn)的轉(zhuǎn)EPSPS基因大豆NZL06-698,受體品種是蒙豆12和區(qū)域?qū)φ沾蠖?001-311-16；安徽種植點(diǎn)的轉(zhuǎn)EPSPS基因大豆ZJU 31,受體品種是華春3號和區(qū)域?qū)φ沾蠖怪悬S13。研究內(nèi)容：測定不同大豆樣品的根際土的理化性質(zhì),大豆植株、種植及根際土的氮含量,根際土的碳、氮循環(huán)相關(guān)酶活,根際土可培養(yǎng)固氮菌數(shù)量,土壤固氮菌群的結(jié)瘤效應(yīng)以及nifH基因豐度表征的土壤固氮菌群總體豐度變化等指標(biāo),分析了轉(zhuǎn)EPSPS基因抗除草劑大豆對土壤固氮菌群的影響。另外還對吉林種植點(diǎn)的轉(zhuǎn)EPSPS基因大豆NZL06-698和受體大豆蒙豆12的根際土壤樣品進(jìn)行了16SrDNA(V4區(qū))擴(kuò)增子的高通量測序,深入分析了轉(zhuǎn)EPSPS基因抗除草劑大豆對土壤微生物群落包括土壤固氮菌群的影響。研究結(jié)果：1.內(nèi)蒙古種植點(diǎn)的實(shí)驗(yàn)數(shù)據(jù)表明,轉(zhuǎn)EPSPS基因抗除草劑大豆NZL06-698與其受體品種蒙古12的根際土壤的N含量、土壤可培養(yǎng)固氮菌數(shù)量以及nifH基因豐度表征的土壤固氮菌群總體豐度均存在顯著性差異,并且表現(xiàn)為降低了土壤固氮菌群的豐度。2.吉林種植點(diǎn)的實(shí)驗(yàn)數(shù)據(jù)表明,轉(zhuǎn)EPSPS基因抗除草劑大豆NZL06-698與其受體品種蒙古12的大豆種子N含量以及根際土壤的N含量、土壤可培養(yǎng)固氮菌數(shù)量以及nifH基因表征的土壤固氮菌群豐度均存在顯著性差異,同時根據(jù)16SrDNA擴(kuò)增子的高通量測序數(shù)據(jù)析可知,種植轉(zhuǎn)EPSPS基因抗除草劑大豆NZL06-698會使根際土壤微生物的物種豐度和均一度更高,但使根際土壤固氮菌群豐度降低。3.安徽種植點(diǎn)的實(shí)驗(yàn)數(shù)據(jù)表明,轉(zhuǎn)EPSPS基因抗除草劑大豆ZJU 31與其受體品種華春3號的種子N含量、土壤固氮酶活、土壤可培養(yǎng)固氮菌數(shù)量均存在顯著性差異,因此種植轉(zhuǎn)EPSPS基因抗除草劑大豆可能會對土壤固氮菌群產(chǎn)生一定的影響,但影響相對較小。對比轉(zhuǎn)基因品系ZJU 31與噴灑一定量草甘膦的轉(zhuǎn)基因品系ZJU 31劑量1數(shù)據(jù),發(fā)現(xiàn)土壤硝酸還原酶、亞硝酸還原酶、土壤固氮酶、蔗糖轉(zhuǎn)化酶活性均存在顯著性差異,因此噴灑一定量的草甘膦可能會對土壤微生物群落產(chǎn)生影響,而且影響相對較大。此結(jié)果還需后續(xù)實(shí)驗(yàn)進(jìn)行驗(yàn)證。結(jié)論：綜合以上數(shù)據(jù)分析可知,種植轉(zhuǎn)EPSPS基因抗除草劑大豆或者噴灑一定量的草甘膦可能會對土壤微生物群落,尤其是對根際土壤固氮菌群產(chǎn)生一定的影響。
[Abstract]:With the development of the plant transgenic technology, the technology has been widely used in the field of agriculture. In 1996, the first commercial anti-weed-resistant transgenic soybean in the United States was approved for planting, and the planting area of the transgenic soybean gradually increased, most of which are anti-herbicide soybeans. The transgenic soybean has brought great economic and social benefits to the human, but the safety of the environment has also gradually raised the concern of people, especially on the ecological environment of the soil. the microorganisms in the soil are very sensitive to external interference, and the accumulation of the foreign genes and the expression products of the transgenic crops in the soil ecosystem is likely to have a certain influence on the microbial community in the soil, In addition, there is a close relationship between the soybean and the nitrogen-fixing-related microbial community in the soil, so in the study of the environmental safety of the soybean, the study on the change of the soil microbial community diversity, in particular the effect of the soil nitrogen-fixing-related microbial community, is very necessary. The selected planting points and the soybean varieties were as follows: the anti-herbicide soybean NZL06-698, the transgenic soybean 12 and the region control soybean black river 43; the transgenic EPSPS gene soybean NZL06-698 in the Jilin planting point; the acceptor variety is the Mongolian bean 12 and the region control soybean 2001-311-16; The transgenic soybean ZJU 31 of Anhui planting point is the soybean ZJU 31 of Huichun No.3 and the region control soybean. The contents of the study are as follows: the physical and chemical properties of the rhizosphere soil of different soybean samples, the nitrogen content of the soybean plant, the planting and the rhizosphere soil, the carbon of the rhizosphere soil, the relative enzyme activity of the nitrogen cycle, the amount of the nitrogen-fixing bacteria in the rhizosphere soil, The effect of the anti-herbicide soybean on the nitrogen-fixing bacteria population in the soil was analyzed by the index of the nodulation effect of the nitrogen-fixing bacteria group and the change of the total abundance of the nitrogen-fixing bacteria in the soil of the nfH gene. In addition, the high-throughput sequencing of the 16SrDNA (V4 region) amplicon was carried out on the rhizosphere soil samples of the transgenic soybean NZL06-698 and the receptor-soybean mask 12 of the Jilin planting site, and the effect of the anti-herbicide soybean on the soil microbial community including the soil nitrogen-fixing bacteria group was analyzed. Study results:1. The experimental data of the planting point in Inner Mongolia showed that there was a significant difference in the N content of the RPSPS gene anti-herbicide soybean NZL06-698 and the rhizosphere soil of the receptor type Mongolia 12, the number of nitrogen-fixing bacteria in the soil, and the total abundance of the soil nitrogen-fixing bacteria characterized by the abundance of the nfH gene. And is characterized in that the abundance of the soil nitrogen-fixing bacteria group is reduced. The experimental data of the planting point of Jilin showed that there was a significant difference in the N content of soybean seed N and the N content of the rhizosphere soil, the number of nitrogen-fixing bacteria in the soil, and the abundance of the nitrogen-fixing bacteria in the soil, which was characterized by the nfH gene. At the same time, according to the high-throughput sequencing data of the 16SrDNA amplicon, the plant-transferring EPSPS gene anti-herbicide soybean NZL06-698 can make the species abundance and the degree of the rhizosphere soil microorganism higher, but the abundance of the nitrogen-fixing bacteria in the rhizosphere is reduced. The experimental data of the planting point in Anhui showed that there was a significant difference between the N content of the anti-herbicide soybean ZJU 31 of the transgenic EPSPS gene and its receptor type Huichun No.3, the activity of the nitrogen-fixing enzyme of the soil and the number of the nitrogen-fixing bacteria in the soil. Therefore, the anti-herbicide soybean planted to the EPSPS gene may have a certain effect on the soil nitrogen-fixing bacteria group, but the effect is relatively small. The results showed that there was a significant difference in the activity of nitrate reductase, nitrite reductase, nitrogen-fixing enzyme and invertase in the transgenic line ZJU 31 and ZJU 31, which was sprayed with a certain amount of plant. Therefore, a certain amount of grass can be sprayed to the soil microbial community, and the effect is relatively large. The results also need to be validated in a follow-up experiment. Conclusion: From the above data analysis, it can be found that the anti-herbicide-resistant soybean or a certain amount of the grass-root of the plant-transferred EPSPS gene may have a certain effect on the soil microbial community, especially the nitrogen-fixing bacteria in the rhizosphere soil.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S154.36;S565.1
本文編號：2490001
[Abstract]:With the development of the plant transgenic technology, the technology has been widely used in the field of agriculture. In 1996, the first commercial anti-weed-resistant transgenic soybean in the United States was approved for planting, and the planting area of the transgenic soybean gradually increased, most of which are anti-herbicide soybeans. The transgenic soybean has brought great economic and social benefits to the human, but the safety of the environment has also gradually raised the concern of people, especially on the ecological environment of the soil. the microorganisms in the soil are very sensitive to external interference, and the accumulation of the foreign genes and the expression products of the transgenic crops in the soil ecosystem is likely to have a certain influence on the microbial community in the soil, In addition, there is a close relationship between the soybean and the nitrogen-fixing-related microbial community in the soil, so in the study of the environmental safety of the soybean, the study on the change of the soil microbial community diversity, in particular the effect of the soil nitrogen-fixing-related microbial community, is very necessary. The selected planting points and the soybean varieties were as follows: the anti-herbicide soybean NZL06-698, the transgenic soybean 12 and the region control soybean black river 43; the transgenic EPSPS gene soybean NZL06-698 in the Jilin planting point; the acceptor variety is the Mongolian bean 12 and the region control soybean 2001-311-16; The transgenic soybean ZJU 31 of Anhui planting point is the soybean ZJU 31 of Huichun No.3 and the region control soybean. The contents of the study are as follows: the physical and chemical properties of the rhizosphere soil of different soybean samples, the nitrogen content of the soybean plant, the planting and the rhizosphere soil, the carbon of the rhizosphere soil, the relative enzyme activity of the nitrogen cycle, the amount of the nitrogen-fixing bacteria in the rhizosphere soil, The effect of the anti-herbicide soybean on the nitrogen-fixing bacteria population in the soil was analyzed by the index of the nodulation effect of the nitrogen-fixing bacteria group and the change of the total abundance of the nitrogen-fixing bacteria in the soil of the nfH gene. In addition, the high-throughput sequencing of the 16SrDNA (V4 region) amplicon was carried out on the rhizosphere soil samples of the transgenic soybean NZL06-698 and the receptor-soybean mask 12 of the Jilin planting site, and the effect of the anti-herbicide soybean on the soil microbial community including the soil nitrogen-fixing bacteria group was analyzed. Study results:1. The experimental data of the planting point in Inner Mongolia showed that there was a significant difference in the N content of the RPSPS gene anti-herbicide soybean NZL06-698 and the rhizosphere soil of the receptor type Mongolia 12, the number of nitrogen-fixing bacteria in the soil, and the total abundance of the soil nitrogen-fixing bacteria characterized by the abundance of the nfH gene. And is characterized in that the abundance of the soil nitrogen-fixing bacteria group is reduced. The experimental data of the planting point of Jilin showed that there was a significant difference in the N content of soybean seed N and the N content of the rhizosphere soil, the number of nitrogen-fixing bacteria in the soil, and the abundance of the nitrogen-fixing bacteria in the soil, which was characterized by the nfH gene. At the same time, according to the high-throughput sequencing data of the 16SrDNA amplicon, the plant-transferring EPSPS gene anti-herbicide soybean NZL06-698 can make the species abundance and the degree of the rhizosphere soil microorganism higher, but the abundance of the nitrogen-fixing bacteria in the rhizosphere is reduced. The experimental data of the planting point in Anhui showed that there was a significant difference between the N content of the anti-herbicide soybean ZJU 31 of the transgenic EPSPS gene and its receptor type Huichun No.3, the activity of the nitrogen-fixing enzyme of the soil and the number of the nitrogen-fixing bacteria in the soil. Therefore, the anti-herbicide soybean planted to the EPSPS gene may have a certain effect on the soil nitrogen-fixing bacteria group, but the effect is relatively small. The results showed that there was a significant difference in the activity of nitrate reductase, nitrite reductase, nitrogen-fixing enzyme and invertase in the transgenic line ZJU 31 and ZJU 31, which was sprayed with a certain amount of plant. Therefore, a certain amount of grass can be sprayed to the soil microbial community, and the effect is relatively large. The results also need to be validated in a follow-up experiment. Conclusion: From the above data analysis, it can be found that the anti-herbicide-resistant soybean or a certain amount of the grass-root of the plant-transferred EPSPS gene may have a certain effect on the soil microbial community, especially the nitrogen-fixing bacteria in the rhizosphere soil.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S154.36;S565.1
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