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林业科学 ›› 2015, Vol. 51 ›› Issue (11): 69-75.doi: 10.11707/j.1001-7488.20151109

• 论文与研究报告 • 上一篇    下一篇

转多基因库安托杨对土壤微生物群落功能的影响

朱文旭, 张冰玉, 黄秦军, 褚延广, 丁昌俊, 张伟溪, 苏晓华   

  1. 林木遗传育种国家重点实验室 中国林业科学研究院林业研究所 北京 100091
  • 收稿日期:2014-04-10 修回日期:2014-06-06 出版日期:2015-11-25 发布日期:2015-12-08
  • 通讯作者: 苏晓华
  • 基金资助:
    国家高技术研究发展计划项目(2011AA100201); 国家林业公益性行业科研专项经费重大项目 (201004004)。

Effects of Multi-Gene Transgenic Populus×euramericana 'Guariento' on the Function of Microbial Population in the Rhizosphere Soil

Zhu Wenxu, Zhang Bingyu, Huang Qinjun, Chu Yanguang, Ding Changjun, Zhang Weixi, Su Xiaohua   

  1. State Key Laboratory of Tree Genetics and Breeding Research Institute of Forestry, CAF Beijing 100091
  • Received:2014-04-10 Revised:2014-06-06 Online:2015-11-25 Published:2015-12-08

摘要: [目的]研究转基因库安托杨和非转基因库安托杨根际土壤微生物的碳源利用水平,为揭示转多基因库安托杨对土壤微生物系统的影响提供科学依据。[方法]在北京市房山区韩村河东营苗圃,取转基因库安托杨5个无性系(D5-9,D5-19,D5-20,D5-21和D5-24)、非转基因库安托杨(CK)根际土壤以及无植物种植土壤(NP)。通过Biolog生态板培养土壤微生物,用多功能酶标仪读数,计算碳源利用能力、丰富度指数、优势度指数和均匀度指数。[结果]温育24 h后,土壤微生物逐渐适应了Biolog微平板基质环境,随后进入对数增长期,平均颜色变化率(AWCD)快速增加直至96 h,此时微生物活性旺盛,碳源开始被明显利用,随后缓慢增长,直至趋于稳定。各培养阶段的AWCD均为D5-24的根际土最高,表明其微生物群落碳源利用量最多,代谢活性最强,CK根际土次之,而NP土微生物代谢最慢,活性最弱。说明栽植杨树的根际土壤丰富了碳源,从而促进微生物的代谢活性。土壤微生物对胺类、酚酸类、羧酸类、多聚物、氨基酸和碳水化合物的利用强度存在差异,微生物对碳水化合物和氨基酸的代谢比较旺盛而对酚酸类和胺类的代谢较弱。对72 h的数据进行主成分(PCA)分析,31个主成分因子中前20个的方差贡献率达100%,其中主成分1的方差贡献率为42.31%,主成分2为13.57%,第3~20主成分贡献率较小,为1.32%~11.64%。PC1和PC2的累积贡献率高达55.88%,相关性分析表明,与PC1正相关程度最高的碳源为D-纤维二糖(0.900),负相关的碳源为衣康酸(-0.266);与PC2正相关程度最高的碳源为苯乙胺(0.668),负相关的碳源为α-丁酮酸(-0.631)。NP和CK分别分布在最小和最大两端。在PC2轴上NP和D5-9分布在正向,D5-20和D5-24分布在负向,CK、D5-19和D5-21两边均有分布,NP与其他无性系之间的分布区域有明显差异。[结论]转多基因库安托杨对土壤微生物系统没有明显的影响。不同转基因无性系之间的差别可能是由于外源基因插入位置、拷贝数等差异导致植株的生理生化及代谢特性发生变化,而非基因表达产物本身的影响。不论栽植非转基因杨树还是转基因杨树都会增加根际土壤微生物的代谢活性。

关键词: 转多基因杨树, 土壤微生物, Biolog

Abstract: [Objective]This paper aims at studying the carbon utilization level of the rhizosphere microbials of transgenic poplars (Populus×euramericana 'Guariento') and non-transgenic poplars, and the results would be helpful in the prediction whether the transgenic poplars have significant adverse effects on the soil microorganism system. [Method]The rhizosphere soil was collected from three different lands:land planted with five strains of transgenic poplars (D5-9,D5-19,D5-20,D5-21 and D5-24), land planted with non-transgenic poplars (CK) and land without any plants(NP) at Hancunhe Dongying nursery in Fangshan District. The ecological plate culture method was used to cultivate soil microbes, multifunction microtiter plate reader was used to calculate the carbon utilization, and the collected data were analyzed with Shannon index, Simpson index and Mclntosh index. [Results]After 24 h incubation, soil microbes gradually adapted to the ecological plate environment, and then entered the logarithmic growth phase, and the arerage well color development (AWCD) rapidly increased until 96 h. A strong microbial activity was observed at this time, and carbon sources began to be obviously used, and then followed by slow growth until stabilized. At each culture stage, the AWCD of D5-24 rhizosphere soil was highest, showing that the amount of carbon source used in the microbial community was the most, and the metabolic activity was the strongest. The non-transgenic rhizosphere was second; and the NP soil microbial had the slowest metabolism and the weakest activity. The result indicated that the rhizosphere soil of poplars was rich in carbon sources, and hence promoted metabolic activity of microorganisms. There were differences in utilization on amines, phenolic acids, carboxylic acids, polymers, amino acids and carbohydrates by soil microorganisms. The carbohydrates and amino acid metabolism were relatively strong while acids and amines utilization was weak. The principal component analysis(PCA) for 72 h data indicated that contribution rate of the first 20 factors was 100% in the 31 principal components of variance. The first main component of a variance had 42.31% contribution rate, the principal component 2 had contribution rate of 13.57 percent, and the third to 20th principal components had smaller contribution, with 1.32%-11.64%. Cumulative contribution rate from PC1 and PC2 was up to 55.88%. Correlation analysis showed that D-cellobiose, the carbon source, had the highest correlation with PC1(0.900), and the carbon source that formed a negative correlation with PC1 was itaconic acid (-0.266). The carbon source that formed the highest positive correlation with PC2 was phenylethylamine (0.668), and the carbon source formed a negative correlation was α-D acid (-0.631). NP and CK were distributed in the minimum and maximum ends, respectively. In the PC2 axis, NP and D5-9 were located in the positive, D5-20 and D5-24 were distributed in the negative, and CK, D5-19 and D5-21 were distributed on both sides. There were significant differences in distribution regions between NP and other clones. [Conclusion]The multi-gene transgenic poplar was not found having significant adverse effects on the soil microorganism system. Differences between genetically clones could be due to the inserted position of exogenous genes, copy number and other differences which lead to changes in physiological and biochemical characteristics of plants and metabolism, not due to the effect of gene expression product itself. Regardless of planting non-transgenic poplar or transgenic poplars can increase the metabolic activity of rhizosphere soil microbes.

Key words: multi-gene transgenic poplar, soil microorganism, Biolog

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