不同生育期转Bt基因棉种植对根际土壤微生物的影响

doi:10.3773/j.issn.1005-264x.2008.01.023

摘要/Abstract

摘要：

在盆栽种植条件下,比较研究了两个转Bt基因棉(Gossypium hirsutum)与对照棉对根际土壤细菌、放线菌、真菌和主要功能类群及多样性的影响差异。结果表明:两个转Bt基因棉根际土壤均可检测到Bt蛋白,且不同转Bt基因棉根系分泌Bt蛋白量以及Bt蛋白在根际土壤中的降解率不同。与各自对照相比,转Bt基因棉对细菌和真菌生长繁殖有促进作用,对放线菌、好气固氮菌和钾细菌数量没有显著影响。苗期和花期转Bt基因棉均可显著提高氨化细菌、显著降低无机溶磷菌数量,花期均可显著提高好气纤维分解菌、显著降低有机溶磷菌数量,‘Bt冀668’苗期也可显著提高好气纤维分解菌数量。转Bt基因棉根际土壤好气纤维分解菌、有机和无机溶磷菌多度发生了变化。尽管功能类群总数转Bt基因棉高于各自对照常规棉,但群落多样性和均匀度都有所下降,优势集中性表现明显,且花期转Bt基因棉多样性参数值以及功能类群数量的变化幅度大于苗期。

关键词: 转Bt基因棉, Bt蛋白, 微生物, 多样性

Abstract:

Aims Ecological risk assessments have paid increasing attention to the field release of transgenic Bt crops. Some studies have reported that Bt protein could enter the soil ecosystem through root exudates, crop residues, crop decomposition and pollen. Protein from transgenic Bt crops is a potential hazard for soil non-target organisms and biodiversity, and this may affect soil ecological function and soil nutrients. This study aims to evaluate the soil ecological risk of transgenic Bt cotton.
Methods A pot experiment with two types of transgenic Bt cotton, ‘BtXincai1’ and ‘BtJi668’, was conducted to study the effects of transgenic Bt cotton planting on rhizosphere soil microorganisms. The plant was carefully removed from the pot at seedling, budding, flowering, bolling and boll-opening stages. Rhizosphere soil approximately 0-5 mm in diameter was separated from bulk soil by gently shaking the root system. Samples were frozen at -20 ℃ after sieving for subsequent Bt protein analyses, or stored at 4 ℃ for microorganism analyses, which were begun within a day of sampling. Bt protein was analyzed by ELISA BOX. The populations of three main microorganism groups and some functional groups were measured by cultivation.
Important findings The content of Bt protein exuded by different transgenic Bt cottons and the degradation rate of Bt protein in different rhizosphere soils were different. Compared with the controls, transgenic Bt cottons seemed to stimulate the reproduction of rhizosphere soil bacteria and fungi. However, no significant effects on the populations of actinomycete, aerobic nitrogen fixing bacteria and kalium-solubilizing bacteria were observed. Transgenic Bt cottons increased the population of ammonifying bacteria and decreased that of inorganic phosphorus-solubilizing bacteria significantly at seedling and flowering stages, whereas they increased the population of aerobic cellulose-decomposing bacteria and decreased that of organic phosphorus-solubilizing bacteria significantly only at the flowering stage. ‘BtJi668’ also enhanced the population of aerobic cellulose-decomposing bacteria significantly at the seedling stage. The abundance of aerobic cellulose-decomposing bacteria and organic and inorganic phosphorus-solubilizing bacteria in transgenic Bt cottons rhizosphere soils changed. Although the total population of functional groups in transgenic Bt cottons rhizosphere soils was higher than in control cultivars, community diversity (H) and evenness (J) decreased and dominance concentration (C) increased. The change range of diversity indexes and population of functional groups in transgenic Bt cottons rhizosphere soils at the flowering stage was higher than at the seedling stage.

Key words: transgenic Bt cotton, Bt protein, microorganisms, diversity

张美俊, 杨武德, 李燕娥. 不同生育期转Bt基因棉种植对根际土壤微生物的影响. 植物生态学报, 2008, 32(1): 197-203. DOI: 10.3773/j.issn.1005-264x.2008.01.023

ZHANG Mei-Jun, YANG Wu-De, LI Yan-E. EFFECTS OF Bt TRANSGENIC COTTONS PLANTING ON RHIZOSPHERE SOIL MICROORGANISMS AT DIFFERENT GROWTH STAGES. Chinese Journal of Plant Ecology, 2008, 32(1): 197-203. DOI: 10.3773/j.issn.1005-264x.2008.01.023

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://www.plant-ecology.com/CN/10.3773/j.issn.1005-264x.2008.01.023

https://www.plant-ecology.com/CN/Y2008/V32/I1/197

图/表 4

参考文献 22

[1]	Angle JS (1994). Release of transgenic plants: biodiversity and population-level consideration. Molecular Ecology, 3, 45-50.
[2]	Bai LX (柏立新), Zhang LW (张龙娃), Chen XB (陈小波), Feng HJ (冯汉金) (2003). Composition and diversity of the weed community in transgenic Bt cotton (four bollgard strains) fields. Acta Phytoecologica Sinica (植物生态学报), 27, 610-616. (in Chinese with English abstract)
[3]	Blackwood CB, Buyer JB (2004). Soil microbial communities associated with Bt and no-Bt corn in three soils. Journal of Environmental Quality, 33, 832-836.
[4]	Crecchio C, stotzky G (1998). Insecticidal activity and biodegradation of the toxin from Bacillus thuringiensis subsp. kurstaki bound to humic acids from soil. Soil Biology and Biochemistry, 30, 463-470.
[5]	Ding ZY (丁志勇), Xu CR (许崇任), Wang RJ (王戎疆) (2001). Comparison of several important isoenzymes between Bt cotton and regular cotton. Acta Ecologica Sinica (生态学报), 21, 332-336. (in Chinese with English abstract)
[6]	Donegan KK, Palm CJ, Fieland VJ, Porteous LA, Ganio LM, Schaller DL, Bucao LQ, Seidler RJ (1995). Changes in levels, species, and DNA fingerprints of soil microorganisms associated with cotton expressing the Bacillus thuringiensis var. kurstaki endotoxin. Applied Soil Ecology, 2, 121-124.
[7]	Escher N, Kach B, Nentwig W (2000). Decomposition of transgenic Bacillus thuringiensis maize by microorganisms and woodlice Porcellio scaber (Crustacea: Isopoda). Basic and Applied Ecology, 1, 161-169.
[8]	Head G, Surber JB, Watson JA, Martin JW, Duan JJ (2002). No detection of Cry1Ac protein in soil after multiple years of transgenic Bt cotton (Bollgard) use. Environmental Entomology, 31, 30-36.
[9]	MacGregor AN, Turner MA (2000). Soil effects of transgenic agriculture: biological processes and ecological consequences. Soil News, 48, 166-169.
[10]	Savka MA, Farrand SK (1997). Modification of rhizobacterial populations by engineering bacterium utilization of a novel plant-produced resource. Nature Biotechnology, 15, 363-368.
[11]	Saxena D, Flores S, Stotzky G (1999). Insecticidal toxin in root exudates from Bt corn. Nature, 402, 480.
[12]	Saxena D, Stotzky G (2000). Insecticidal toxin from Bacillus thuringiensis is released from roots of transgenic Bt corn in vitro and in situ. FEMS Journal Microbiology Ecology, 33, 35-39.
[13]	Saxena D, Stotzky G (2001). Bacillus thuringiensis (Bt) toxin released from root exudates and biomass of Bt corn has no apparent effect on earthworms, nematodes, protozoa, bacteria, and fungi in soil. Soil Biology and Biochemistry, 33, 1225-1230.
[14]	Shen FF (沈法富), Han XL (韩秀兰), Fan SL (范术丽) (2004). Changes in microbial flora and bacterial physiological group diversity in rhizosphere soil of transgenic Bt cotton. Acta Ecologica Sinica (生态学报), 24, 432-437. (in Chinese with English abstract)
[15]	Sun CX (孙彩霞), Chen LJ (陈利军), Wu ZJ (武志杰) (2004). Persistence of Bt toxin in soil and its effects on soil phosphatase activity. Acta Pedologica Sinica (土壤学报), 41, 761-765. (in Chinese with English abstract)
[16]	Tapp H, Stotzky G (1995). Dot blot enzyme-linked immunosorbent assay for monitoring the fate of insecticidal toxins from Bacillus thuringiensis in soil. Applied and Environmental Microbiology, 61, 602-609.
[17]	Tapp H, Stotzky G (1998). Presistance of the insecticidal toxin from Bacillus thuringiensis subsp. kurstaki in soil. Soil Biology and Biochemistry, 30, 471-476.
[18]	Teng Y (滕应), Huang CY (黄昌勇) (2002). Ecological effect of heavy metals on soil microbes and research advances on the mechanisms of bioremediation. Soil and Environmental Sciences (土壤与环境), 11, 85-89. (in Chinese with English abstract)
[19]	Trevors JT, Kuikman P, Watson B (1994). Transgenic plants and biogeochemical cycles. Molecular Ecology, 3, 57-64.
[20]	Watrud LS, Seidler RJ (1998). Nontarget ecological effects of plants, microbial, and chemical introductions to terrestrial system. In: Huang PM ed. Soil Chemistry and Ecosystem Health, Special Publication 52. Soil Science Society of America, Madison, Wiscomisin, 313-340.
[21]	Xu GH (许光辉), Zheng HY (郑洪元) (1986). Handbook of Methods for Edaphon Analysis (土壤微生物分析方法手册). China Agriculture Press, Beijing, 102-135. (in Chinese)
[22]	Zheng Y (郑宇), Li YR (李月仁), Hu QY (胡奇勇), Li BJ (李本金), Hu XB (胡习斌) (2003). Analysis impacts of soil micro-biotic community by transgenic crops and engineered microorganisms. Acta Agriculture Universitatis Jiangxiensis (江西农业大学学报), 25, 67-70.

微生物 Microorganisms	处理 Treatments	苗期 Seedling	蕾期 Budding	花期 Flowering	盛铃期 Bolling	吐絮期 Boll-opening
细菌Bacteria (×10⁶ cfu·g^-1)	‘Bt新彩1’BtXincai1	93.30±2.63^b	418.60±3.77^a	1078.63±9.70^b	749.28±4.77^b	356.20±7.11^b
	‘新彩1’Xincai1	92.77±4.54^b	237.87±8.24^b	678.10±9.46^d	466.50±7.34^d	251.10±8.55^c
	‘Bt冀668’BtJi668	116.10±7.08^a	433.62±6.76^a	1209.73±8.04^a	884.27±6.34^a	457.73±5.51^a
	‘冀668’Ji668	114.57±7.78^a	219.70±9.37^c	828.20±7.59^c	528.18±6.26^c	237.77±6.09^c
放线菌Actinomycete (×10⁵ cfu·g^-1)	‘Bt新彩1’BtXincai1	20.83±4.57	28.34±9.17	34.39±2.51	31.14±4.97	19.90±2.04
	‘新彩1’Xincai1	24.26±2.14	26.36±2.33	35.41±4.04	32.46±5.63	18.90±3.21
	‘Bt冀668’BtJi668	21.27±5.11	22.81±5.28	34.94±9.19	32.85±5.47	21.30±1.15
	‘冀668’Ji668	25.72±3.14	26.19±1.17	35.04±3.68	31.87±6.68	20.81±2.69
真菌Fungi (×10³ cfu·g^-1)	‘Bt新彩1’BtXincai1	4.32±0.48	11.57±1.15^ab	28.50±1.97^a	15.25±1.23^a	7.24±0.42^b
	‘新彩1’Xincai1	4.23±0.56	10.77±1.01^bc	18.76±1.10^c	9.17±0.20^b	6.86±0.79^ab
	‘Bt冀668’BtJi668	4.21±0.49	13.15±1.46^a	25.74±1.90^b	16.42±1.16^a	9.58±1.00^a
	‘冀668’Ji668	4.14±0.42	9.24±1.01^c	16.53±1.16^c	10.27±0.32^b	6.18±0.87^b

微生物 Microorganisms	处理 Treatments	苗期 Seedling	蕾期 Budding	花期 Flowering	盛铃期 Bolling	吐絮期 Boll-opening
细菌Bacteria (×10⁶ cfu·g^-1)	‘Bt新彩1’BtXincai1	93.30±2.63^b	418.60±3.77^a	1078.63±9.70^b	749.28±4.77^b	356.20±7.11^b
	‘新彩1’Xincai1	92.77±4.54^b	237.87±8.24^b	678.10±9.46^d	466.50±7.34^d	251.10±8.55^c
	‘Bt冀668’BtJi668	116.10±7.08^a	433.62±6.76^a	1209.73±8.04^a	884.27±6.34^a	457.73±5.51^a
	‘冀668’Ji668	114.57±7.78^a	219.70±9.37^c	828.20±7.59^c	528.18±6.26^c	237.77±6.09^c
放线菌Actinomycete (×10⁵ cfu·g^-1)	‘Bt新彩1’BtXincai1	20.83±4.57	28.34±9.17	34.39±2.51	31.14±4.97	19.90±2.04
	‘新彩1’Xincai1	24.26±2.14	26.36±2.33	35.41±4.04	32.46±5.63	18.90±3.21
	‘Bt冀668’BtJi668	21.27±5.11	22.81±5.28	34.94±9.19	32.85±5.47	21.30±1.15
	‘冀668’Ji668	25.72±3.14	26.19±1.17	35.04±3.68	31.87±6.68	20.81±2.69
真菌Fungi (×10³ cfu·g^-1)	‘Bt新彩1’BtXincai1	4.32±0.48	11.57±1.15^ab	28.50±1.97^a	15.25±1.23^a	7.24±0.42^b
	‘新彩1’Xincai1	4.23±0.56	10.77±1.01^bc	18.76±1.10^c	9.17±0.20^b	6.86±0.79^ab
	‘Bt冀668’BtJi668	4.21±0.49	13.15±1.46^a	25.74±1.90^b	16.42±1.16^a	9.58±1.00^a
	‘冀668’Ji668	4.14±0.42	9.24±1.01^c	16.53±1.16^c	10.27±0.32^b	6.18±0.87^b

生育期 Growth Stages	处理 Treatments	好气固氮菌 Aerobic nitrogen fixing bacteria	氨化细菌 Ammonifying bacteria	好气纤维分解菌 Aerobic cellulose- decomposing bacteria	有机溶磷菌 Organic phosphorus- solubilizing bacteria	无机溶磷菌 Inorganic phosphorus- solubilizing bacteria	钾细菌 Kalium- solubilizing bacteria
苗期	‘Bt新彩1’BtXincai1	0.16±0.02	42.17±2.89^b	14.10±1.72^ab	0.99±0.10	10.58±1.16^b	14.74±0.79^ab
Seedling	‘新彩1’Xincai1	0.15±0.02	30.37±0.74^c	12.31±1.19^b	1.08±0.09	16.92±2.68^a	13.29±2.10^a
	‘Bt冀668’BtJi668	0.14±0.03	57.27±3.67^a	14.55±1.18^a	1.12±0.12	12.25±1.50^b	17.53±1.03^a
	‘冀668’Ji668	0.15±0.01	45.30±2.62^b	8.74±0.37^c	1.16±0.20	18.78±2.46^a	16.70±1.95^ab
花期	‘Bt新彩1’BtXincai1	0.70±0.17	95.43±7.67^b	35.81±1.78^b	2.51±0.57^d	16.34±3.09^c	31.49±0.97
Flowering	‘新彩1’Xincai1	0.81±0.14	51.93±2.38^d	25.25±2.22^c	7.49±0.59^b	28.23±2.00^b	32.75±2.08
	‘Bt冀668’BtJi668	0.75±0.14	111.53±3.47^a	46.42±2.29^a	3.50±0.58^c	21.65±2.53^c	33.47±2.09
	‘冀668’Ji668	0.87±0.09	63.30±4.13^c	23.48±1.61^c	8.48±0.54^a	35.82±2.29^a	32.99±2.26

生育期 Growth Stages	处理 Treatments	好气固氮菌 Aerobic nitrogen fixing bacteria	氨化细菌 Ammonifying bacteria	好气纤维分解菌 Aerobic cellulose- decomposing bacteria	有机溶磷菌 Organic phosphorus- solubilizing bacteria	无机溶磷菌 Inorganic phosphorus- solubilizing bacteria	钾细菌 Kalium- solubilizing bacteria
苗期	‘Bt新彩1’BtXincai1	0.16±0.02	42.17±2.89^b	14.10±1.72^ab	0.99±0.10	10.58±1.16^b	14.74±0.79^ab
Seedling	‘新彩1’Xincai1	0.15±0.02	30.37±0.74^c	12.31±1.19^b	1.08±0.09	16.92±2.68^a	13.29±2.10^a
	‘Bt冀668’BtJi668	0.14±0.03	57.27±3.67^a	14.55±1.18^a	1.12±0.12	12.25±1.50^b	17.53±1.03^a
	‘冀668’Ji668	0.15±0.01	45.30±2.62^b	8.74±0.37^c	1.16±0.20	18.78±2.46^a	16.70±1.95^ab
花期	‘Bt新彩1’BtXincai1	0.70±0.17	95.43±7.67^b	35.81±1.78^b	2.51±0.57^d	16.34±3.09^c	31.49±0.97
Flowering	‘新彩1’Xincai1	0.81±0.14	51.93±2.38^d	25.25±2.22^c	7.49±0.59^b	28.23±2.00^b	32.75±2.08
	‘Bt冀668’BtJi668	0.75±0.14	111.53±3.47^a	46.42±2.29^a	3.50±0.58^c	21.65±2.53^c	33.47±2.09
	‘冀668’Ji668	0.87±0.09	63.30±4.13^c	23.48±1.61^c	8.48±0.54^a	35.82±2.29^a	32.99±2.26

特征参数 Parameter of trait	功能类群 Functional groups	苗期 Seedling				花期 Flowering
特征参数 Parameter of trait	功能类群 Functional groups	‘Bt新彩1’ BtXincai1	‘新彩1’ Xincai1	‘Bt冀668’ BtJi668	‘冀668’ Ji668	‘Bt新彩1’ BtXincai1	‘新彩1’ Xincai1	‘Bt冀668’ BtJi668	‘冀668’ Ji668
多度 Abundance (P_i)	好气固氮菌 Aerobic nitrogen fixing bacteria	0.002	0.002	0.001	0.002	0.004	0.006	0.003	0.006
	氨化细菌 Ammonifying bacteria	0.510	0.410	0.557	0.499	0.523	0.355	0.513	0.383
	好气纤维分解菌 Aerobic cellulose-decomposing bacteria	0.170	0.166	0.142	0.095	0.131	0.172	0.213	0.143
	有机溶磷菌Organic phosphorus-solubilizing bacteria	0.012	0.015	0.011	0.013	0.080	0.051	0.016	0.051
	无机溶磷菌Inorganic phosphorus- solubilizing bacteria	0.128	0.228	0.119	0.207	0.090	0.193	0.101	0.217
	钾细菌 Kalium-solubilizing bacteria	0.178	0.179	0.170	0.184	0.173	0.224	0.154	0.200
总数量 Total individual No. (N)		84.74	74.11	102.86	90.83	182.29	146.47	217.33	164.93
群落多样性 Community diversity (H)		1.279	1.384	1.164	1.277	1.309	1.506	1.274	1.483
均匀度 Evenness (J)		0.714	0.772	0.650	0.713	0.730	0.840	0.711	0.828
优势集中性 Concentration (C)		0.337	0.280	0.373	0.335	0.335	0.246	0.342	0.257