转EPSPS基因抗除草剂大豆对土壤原核微生物群落生态影响的研究
发布时间：2019-05-31 20:38
【摘要】：随着植物转基因技术的发展,该技术在农业领域中得到广泛应用。1996年,美国的第一例商业化的抗草甘膦转基因大豆被批准种植,从此转基因大豆的种植面积也逐渐增加,其中绝大多数是抗除草剂大豆。转基因大豆给人类带来了巨大经济效益和社会效益,但是其对环境的安全性问题也逐渐引起了人们的关注,尤其是对土壤生态环境的影响。土壤中的微生物对外界干扰反应非常灵敏,转基因作物的外源基因及其表达产物在土壤生态系统中的累积很可能会对土壤中的微生物群落存在一定程度的影响,而且大豆与土壤中的固氮相关微生物群落关系密切,因此在评价其环境安全性的研究中,对土壤微生物群落多样性变化的研究,特别是对土壤固氮相关微生物群落的影响是非常必要的。本研究选取的种植点及大豆品种分别为：内蒙古种植点的转EPSPS基因抗除草剂大豆NZL06-698,受体品种是蒙豆12和区域对照大豆黑河43；吉林种植点的转EPSPS基因大豆NZL06-698,受体品种是蒙豆12和区域对照大豆2001-311-16；安徽种植点的转EPSPS基因大豆ZJU 31,受体品种是华春3号和区域对照大豆中黄13。研究内容：测定不同大豆样品的根际土的理化性质,大豆植株、种植及根际土的氮含量,根际土的碳、氮循环相关酶活,根际土可培养固氮菌数量,土壤固氮菌群的结瘤效应以及nifH基因丰度表征的土壤固氮菌群总体丰度变化等指标,分析了转EPSPS基因抗除草剂大豆对土壤固氮菌群的影响。另外还对吉林种植点的转EPSPS基因大豆NZL06-698和受体大豆蒙豆12的根际土壤样品进行了16SrDNA(V4区)扩增子的高通量测序,深入分析了转EPSPS基因抗除草剂大豆对土壤微生物群落包括土壤固氮菌群的影响。研究结果：1.内蒙古种植点的实验数据表明,转EPSPS基因抗除草剂大豆NZL06-698与其受体品种蒙古12的根际土壤的N含量、土壤可培养固氮菌数量以及nifH基因丰度表征的土壤固氮菌群总体丰度均存在显著性差异,并且表现为降低了土壤固氮菌群的丰度。2.吉林种植点的实验数据表明,转EPSPS基因抗除草剂大豆NZL06-698与其受体品种蒙古12的大豆种子N含量以及根际土壤的N含量、土壤可培养固氮菌数量以及nifH基因表征的土壤固氮菌群丰度均存在显著性差异,同时根据16SrDNA扩增子的高通量测序数据析可知,种植转EPSPS基因抗除草剂大豆NZL06-698会使根际土壤微生物的物种丰度和均一度更高,但使根际土壤固氮菌群丰度降低。3.安徽种植点的实验数据表明,转EPSPS基因抗除草剂大豆ZJU 31与其受体品种华春3号的种子N含量、土壤固氮酶活、土壤可培养固氮菌数量均存在显著性差异,因此种植转EPSPS基因抗除草剂大豆可能会对土壤固氮菌群产生一定的影响,但影响相对较小。对比转基因品系ZJU 31与喷洒一定量草甘膦的转基因品系ZJU 31剂量1数据,发现土壤硝酸还原酶、亚硝酸还原酶、土壤固氮酶、蔗糖转化酶活性均存在显著性差异,因此喷洒一定量的草甘膦可能会对土壤微生物群落产生影响,而且影响相对较大。此结果还需后续实验进行验证。结论：综合以上数据分析可知,种植转EPSPS基因抗除草剂大豆或者喷洒一定量的草甘膦可能会对土壤微生物群落,尤其是对根际土壤固氮菌群产生一定的影响。
[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.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.36;S565.1
本文编号：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.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S154.36;S565.1
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相关期刊论文 前2条
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