张富丽,
尹全,
毛建霏,
郭灵安,
雷绍荣,
刘文娟,
罗苹,
王东,
宋君,
杨晓凤^,
四川省农业科学院分析测试中心 成都 610066
基金项目: 四川省农业科学院论文基金项目2016LWJJ-010
四川省创新能力提升工程公益深化项目2016GYSH-032

详细信息

作者简介:张富丽, 长期从事转基因成分检测与环境影响评价分析研究。E-mail:zhang_fannie@163.com

通讯作者:杨晓凤, 主要研究方向为农产品质量控制及安全评价。E-mail:yangxiaofeng_cd@sina.com

中图分类号:S1

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出版历程

收稿日期:2019-12-19
录用日期:2020-02-04
刊出日期:2020-05-01

Effects of Bacillus thuringiensis transgenic cotton straw returning to field on soil fertility

ZHANG Fuli,
YIN Quan,
MAO Jianfei,
GUO Ling'an,
LEI Shaorong,
LIU Wenjuan,
LUO Ping,
WANG Dong,
SONG Jun,
YANG Xiaofeng^,
Analysis and Determination Center, Sichuan Academy of Agricultural Sciences, Chengdu 610066, China
Funds: the Dissertation Fund of Sichuan Academy of Agricultural Sciences2016LWJJ-010
the Special Funds of Ability Promotion from Sichuan Provincial Finance2016GYSH-032

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Corresponding author:YANG Xiaofeng, E-mail:yangxiaofeng_cd@sina.com

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摘要
摘要:随着转基因的快速发展，大量转Bt棉秸秆的合理利用和处理是不可忽视的重要课题之一。为明确Bt棉秸秆还田利用的可行性和安全性，本研究以不同抗虫转Bt基因棉和常规棉花‘泗棉3号’为研究材料，在分别种植1、2年后将秸秆机械粉碎后原位还田，测试土壤中Bt蛋白残留量、土壤酶活性及养分含量的变化，分析Bt棉秸秆原位还田对土壤肥力特性的影响。研究结果表明，秸秆还田40 d后，Bt棉样地土壤中Bt残留蛋白检测值较低，均与非转基因棉样地无显著性差异。棉秸秆还田后，土壤脲酶、蔗糖酶、蛋白酶、多酚氧化酶、过氧化氢酶、碱性磷酸酶活性皆较秸秆还田前增加，但土壤纤维素酶活性较之前降低。棉秸秆还田使土壤中有机质、有效磷、碱解氮、速效钾和全氮等养分含量及pH明显增加，而Bt抗虫棉与常规棉秸秆还田后对土壤肥力的影响不存在显著差异。对土壤综合肥力指数评价结果表明，秸秆还田对土壤肥力提升与Bt棉抗虫水平无关，土壤肥力指数在两年间由Ⅲ级水平上升至Ⅱ级水平。综上，Bt棉花秸秆还田不会造成土壤综合肥力降低，相反能有效提升土壤肥力；同时还田利用措施可对转基因植株有效灭活，与转基因植物秸秆利用和无害化处理要求相契合。生产中用于Bt转基因棉花秸秆利用和处理在一定程度上是安全可行的。
关键词:转Bt基因棉/
秸秆还田/
转基因植物/
Bt蛋白残留/
土壤肥力
Abstract:With the rapid development of genetically modified organisms, how to use and deal with a large amount of Bacillus thuringiensis (Bt) transgenic cotton straw is an important issue. To study and clarify the feasibility and safety of straw incorporation of transgenic Bt cotton into the field, four kinds of Bt cotton with different levels of insect resistance and one non-transgenic conventional cotton (Simian no. 3) were analyzed. After one or two years of planting, all the cotton straws were mechanically crushed and returned to the field in situ. The content of residual Bt protein, activities of several soil enzymes, and nutrient contents in soil were detected and the changes were further analyzed. The effects of Bt cotton straw returning on nutrient characteristics and fertility of soil were analyzed. The Bt protein residues in soils were very few and almost undetectable after 40 days of straw recycling of transgenic Bt cotton with different insect resistance levels, and there was no significant differences between the transgenic field plots and non-transgenic ones. After cotton straws were returned into the field, the activities of urease, sucrase, protease, polyphenol oxidase, catalase, and alkaline phosphatase in soil obviously increased, while the activity of cellulose did not increase. The activity of soil cellulase was lower than that before cotton planting. The amounts of organic matter, available phosphorus, available nitrogen, available potassium, and total nitrogen in soil increased remarkably both for the first year and for the second year of cotton planting, and so did the pH value. Similarly, there were not any significant differences in variations of all nutrient content between Bt transgenic treatments and non-transgenic ones. The soil comprehensive fertility index resulted by straw recycling was calculated and the effect on the fertility suitability was evaluated. The fertility index rose from level Ⅲ to level Ⅱ in two years, which was not related to the insect resistance level of Bt transgenic cotton. The soil fertility was not adversely decreased but increased, in fact, by straw returning treatment of Bt cottons. Returning Bt cotton straw to the field could effectively improve the soil fertility index. In addition, this straw recycling scheme could effectively kill transgenic plants, and would meet the disposal requirements of transgenic plants. The collective data confirmed that returning treatment, to a certain extent, should be feasible and safe for dealing with mass residues of Bt transgenic cotton in practice.
Key words:Bacillus thuringiensis transgenic (Bt) cotton/
Straw returning/
Transgenic plant/
Residual Bt protein/
Soil fertility

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表1转Bt基因棉秸秆还田前后土壤养分含量特征性统计
Table1.Descriptive statistics of soil nutrients contents before and after recycling of transgenic Bt cotton straw

土层 Soil layer	指标 Soil index	取样时期 Sampling period	均值±标准差 Mean±S.D.	变幅 Amplitude	变异系数 Variation coefficient (%)
耕作层 Plough layer	有机质 Organic matter (g?kg–1)	0	17.83±1.54a	14.43~20.69	8.64
		Ⅰ	22.27±1.14b	20.50~23.98	5.12
		Ⅱ	29.8±0.92c	28.58~31.34	3.09
	有效磷 Available P (mg·kg–1)	0	9.09±5.19a	2.30~17.8	57.10
		Ⅰ	54.13±4.02b	49.61~64.41	7.43
		Ⅱ	82.21±3.68c	76.52~86.70	4.48
	速效钾 Available K (mg·kg–1)	0	63.42±13.79a	46.03~99.00	21.74
		Ⅰ	100.83±8.32b	86.44~113.94	8.25
		Ⅱ	136.77±10.32c	119.99~150.88	7.55
	碱解氮 Alkali-hydrolysis N (mg·kg–1)	0	104.64±6.57a	93.19~114.97	6.28
		Ⅰ	118.87±8.00b	103.41~132.02	6.73
		Ⅱ	130.83±5.73c	118.66~139.63	4.38
	全氮 Total N (g·kg–1)	0	1.02±0.05a	0.94~1.09	4.90
		Ⅰ	1.16±0.07b	1.03~1.26	6.03
		Ⅱ	1.30±0.08c	1.12~1.45	6.15
耕作层 Plough layer	pH值 pH value	0	5.98±0.36a	5.56~6.64	6.02
		Ⅰ	6.31±0.32b	5.73~6.91	5.07
		Ⅱ	6.70±0.25c	6.25~7.05	3.73
犁底层 Plow pan	有机质 Organic matter (g?kg–1)	0	23.01±1.57a	20.55~26.75	6.85
		Ⅰ	24.80±1.45b	22.05~27.82	5.83
		Ⅱ	27.94±2.59c	24.05~31.64	9.27
	有效磷 Available P (mg·kg–1)	0	7.61±2.79a	4.25~12.36	36.70
		Ⅰ	20.13±1.96b	17.22~24.55	9.72
		Ⅱ	25.39±3.64c	20.23~32.19	14.34
	速效钾 Available K (mg·kg–1)	0	53.18±6.42a	45.23~69.28	12.07
		Ⅰ	63.65±8.39b	52.86~79.85	13.18
		Ⅱ	76.54±8.91c	59.67~95.89	11.64
	碱解氮 Alkali-hydrolysis N (mg·kg–1)	0	138.90±13.82a	109.20~156.25	9.95
		Ⅰ	156.20±14.08b	125.79~181.46	9.01
		Ⅱ	164.80±13.85c	138.31~195.52	8.40
	全氮 Total N (g·kg–1)	0	1.49±0.08a	1.33~1.61	5.57
		Ⅰ	1.66±0.08b	1.56~1.85	5.02
		Ⅱ	1.76±0.11c	1.56~2.01	6.36
	pH	0	6.07±0.23a	5.59~6.45	3.80
		Ⅰ	6.39±0.20b	5.95~6.63	3.19
		Ⅱ	6.58±0.27c	6.02~7.03	4.13
取样时期0、Ⅰ和Ⅱ分别指棉花种植前1年、棉花种植1年秸秆还田后和棉花种植2年秸秆还田后。The sampling periods 0, Ⅰ and Ⅱ are one year before planting cotton, after straw incorporation of the first year and the second year of cotton planting, respectively.

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表2不同抗虫性转基因棉还田40 d后土壤Bt蛋白残留量
Table2.Contents of residual Bt protein in soil after 40 days of straw incorporation of transgenic Bt cotton with different insect resistance levels

土层 Soil layer	抗虫性 Insect resistance level	棉花种植前 Before planting cotton				棉花种植1年秸秆还田后 After straw incorporation of the first year of cotton planting				棉花种植2年秸秆还田后 After straw incorporation of the second year of cotton planting
		Bt蛋白量 Bt protein content (μg·kg–1)		与Cn间差异(P值) Significance of difference with Cn (P value)		Bt蛋白量 Bt protein content (μg·kg–1)		与Cn间差异(P值) Significance of difference with Cn (P value)		Bt蛋白量 Bt protein content (μg·kg–1)		与Cn间差异(P值) Significance of difference with Cn (P value)
耕作层 Plough layer	H	1.220±0.020		0.837		1.853±0.073		0.336		1.806±0.113		0.457
	M	1.180±0.009		0.904		1.590±0.139		0.594		1.718±0.196		0.309
	L	1.196±0.083		0.997		1.642±0.317		0.812		1.760±0.042		0.329
	Cp	1.219±0.069		0.908		1.600±0.236		0.634		2.126±0.063		0.367
	Cn	1.242±0.005		/		1.683±0.039		/		2.117±0.097		/
犁底层 Plow pan	H	1.167±0.018		0.858		1.762±0.245		0.874		1.990±0.444		0.854
	M	1.189±0.062		0.932		1.79±0.002		0.779		1.789±0.035		0.494
	L	1.164±0.059		0.848		1.44±0.035		0.223		2.132±0.385		0.426
	Cp	1.180±0.052		0.900		1.661±0.035		0.782		1.987±0.051		0.866
	Cn	1.209±0.029		/		1.725±0.075		/		1.947±0.012		/
H、M、L代表抗虫性水平分别为高、中、低的非商业化转Bt基因棉花品种; Cp是商业化转Bt基因棉‘GK19’, Cn是非转基因棉品种‘泗棉3号’。H, M and L represent non-commercial Bt cotton varieties with high, medium and low insect resistance levels, respectively. Cp is the commercial Bt cotton variety ‘GK19’, Cn is the non-transgenic cotton variety ‘Simian No. 3’.

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表3不同抗虫性转Bt基因棉花秸秆还田对土壤酶活性的影响
Table3.Effect of straw incorporation of transgenic Bt cotton with different insect resistance levels on soil enzymes activities

土壤酶 Soil enzyme	抗虫性 Insect resistance level	棉花种植前1年 One year before planting cotton	棉花种植1年秸秆还田后 After straw incorporation of the first year of cotton planting				棉花种植2年秸秆还田后 After straw incorporation of the second year of cotton planting
			酶活性 Enzyme activity		变化幅度 Change scale (%)		酶活性 Enzyme activity		变化幅度 Change scale (%)
脲酶 Urease (IU·g–1)	H	186.82±5.10a	198.53±5.17ac		+6.27		208.66±14.74bc		+11.62
	M	157.13±5.46a	190.04±6.54bc		+21.05		197.13±5.98c		+25.58
	L	150.88±8.34a	191.00±8.97b		+26.71		213.25±15.51c		+41.29
	Cp	175.08±1.15a	187.79±2.11b		+7.27		219.13±5.00c		+25.16
	Cn	144.74±3.50a	158.25±5.02b		+9.34		180.75±1.90c		+24.93
蔗糖酶 Sucrase (U·g–1)	H	147.64±6.18a	161.49±3.91b		+9.52		172.15±9.16c		+16.88
	M	133.30±5.85a	162.58±8.14b		+22.19		174.52±5.22c		+31.20
	L	132.52±6.13a	155.85±2.08b		+17.73		166.92±8.89c		+26.20
	Cp	131.93±2.30a	163.26±5.28bc		+23.74		168.16±6.71c		+27.44
	Cn	131.93±2.30a	146.87±9.55b		+11.32		162.71±2.94c		+23.35
蛋白酶 Protease (IU·g–1)	H	19.25±1.45a	22.33±0.67bc		+16.59		23.87±0.50c		+24.53
	M	14.01±0.12a	23.23±0.84bc		+65.82		24.77±0.40c		+76.83
	L	15.82±1.08a	21.38±0.75b		+35.81		23.90±1.42c		+51.85
	Cp	17.87±2.73a	20.64±1.73b		+16.55		24.28±0.68c		+37.72
	Cn	17.87±2.73a	20.39±0.58b		+15.70		23.00±0.47c		+30.57
多酚氧化酶 Polyphenol oxidase (IU·g–1)	H	47.12±4.74a	65.63±5.39b		+40.79		78.29±7.07c		+67.22
	M	30.45±4.13a	64.52±1.17b		+114.77		71.06±2.49c		+135.78
	L	36.95±4.80a	60.87±1.78b		+66.73		72.28±6.33c		+96.37
	Cp	45.62±2.75a	71.33±1.22b		+56.64		80.11±0.30c		+76.07
	Cn	45.62±2.75a	62.42±1.65b		+37.30		72.34±2.04c		+59.08
纤维素酶 Cellulose (U·g–1)	H	22.58±0.74a	16.55±1.41b		-26.57		12.47±1.36c		-44.88
	M	21.53±0.10a	18.80±1.40b		-12.69		16.26±1.61c		-24.52
	L	21.65±0.59a	19.89±0.75a		-8.02		12.46±1.87b		-42.59
	Cp	21.34±1.18a	17.54±1.88b		-17.36		13.36±0.54c		-37.16
	Cn	21.34±1.18a	16.94±1.23b		-20.47		13.22±0.60c		-37.86
过氧化氢酶 Catalase (IU·g–1)	H	31.68±3.04a	37.47±0.65b		+19.08		41.64±1.62c		+32.58
	M	31.05±1.69a	37.23±2.95b		+20.14		44.77±4.68c		+43.98
	L	31.22±1.57a	37.17±2.10bc		+19.42		39.86±2.56c		+28.17
	Cp	31.29±1.49a	39.64±0.75bc		+26.82		42.17±0.73c		+34.93
	Cn	31.29±1.49a	39.70±1.53bc		+26.98		42.01±0.73c		+34.54
碱性磷酸酶 Alkaline Phosphatase (IU·g–1)	H	8.27±0.81a	9.04±0.58a		+9.52		11.54±1.01b		+40.99
	M	6.25±0.14a	8.61±0.58b		+38.05		11.18±1.19c		+78.82
	L	5.42±0.41a	6.54±0.50b		+20.84		9.50±0.21c		+76.10
	Cp	7.00±0.02a	8.95±0.03bc		+27.89		9.71±0.45c		+38.71
	Cn	7.00±0.02a	8.03±0.12b		+14.66		9.69±0.40c		+38.52
H、M、L代表抗虫性水平分别为高、中、低的非商业化转Bt基因棉花品种; Cp是商业化转Bt基因棉‘GK19’, Cn是非转基因棉品种‘泗棉3号’。同行不同小写字母表示3个时期间差异显著(P < 0.05)。H, M and L represent non-commercial Bt cotton varieties with high, medium and low insect resistance levels, respectively. Cp is the commercial Bt cotton variety ‘GK19’, Cn is the non-transgenic cotton variety ‘Simian No. 3’. Different lowercase letters in the same line mean significant differences among three periods at P < 0.05 level.

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表4转Bt基因棉秸秆还田后不同时间土壤酶活变化与棉花抗虫性相关性分析
Table4.Spearman correlation analysis between change of soil enzymes activities at different times after straw incorporation of transgenic Bt cotton and insect-resistance of transgenic cotton

酶 Enzyme	棉花种植1年秸秆还田后 After straw incorporation of the first year of cotton planting				棉花种植2年秸秆还田后 After straw incorporation of the second year of cotton planting
	r		显著性Significance		r		显著性Significance
脲酶Urease	0.083		ns		0.436		ns
蔗糖酶Sucrase	0.131		ns		0.076		ns
蛋白酶Protease	0.316		ns		0.142		ns
多酚氧化酶Polyphenol oxidase	0.218		ns		0.404		ns
纤维素酶Cellulose	0.109		ns		0.142		ns
过氧化氢酶Catalase	0.273		ns		0.131		ns
碱性磷酸酶Alkaline Phosphatase	0.120		ns		0.175		ns
ns表示相关性不显著。“ns” means not significant correlation.

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表5土壤养分评价指标分级标准
Table5.Grading and evaluation criteria of soil nutrients contents

分值 Score	有机质 Organic matter (g·kg–1)	碱解氮 Alkali-hydrolysis N (mg·kg–1)	有效磷 Available P (mg·kg–1)	速效钾 Available K (mg·kg–1)	全氮 Total N (g·kg–1)	pH
100	≥40	≥150	≥40.0	≥200	≥2.0	6.0~7.5
90	30~40	120~150	20~40	150~200	1.5~2.0	5.5~6.0; 7.5~8.0
80	20~30	90~120	10~20	100~150	1.0~1.5	5.0~5.5; 8.0~9.0
70	10~20	60~90	5~10	50~100	0.75~1.0	4.5~5.0; 9.0~9.5
60	5~10	30~60	3~5	30~50	0.5~0.75	4.5~5.0
50	< 10	< 30	< 3	< 30	< 0.5	< 4.5

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表6土壤养分指标权重及其隶属函数类型和阈值
Table6.Weight values, distribution types and infection points of soil nutrient indexes

指标 Index	函数类型 Distribution type	转折点值Infection point				权重 Weight value
		X1	X2	X3	X4
pH	抛物线Parabola type	4.5	6.0	7.5	9.0	0.370
有机质Organic matter (g·kg–1)	S形S type	10	—	—	40	0.247
碱解氮Alkali-hydrolysis N (mg·kg–1)	S形S type	30	—	—	150	0.156
速效磷Available P (mg·kg–1)	S形S type	3	—	—	40	0.095
速效钾Available K (mg·kg–1)	S形S type	30	—	—	200	0.087
全氮Total N (g·kg–1)	S形S type	0.5	—	—	2.0	0.046

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表7不同抗虫性转Bt基因棉花秸秆还田后土壤养分含量及肥力指数的变化
Table7.Changes of soil nutrients contents contents and fertility index after straw incorporation of transgenic Bt cotton with different insect resistance levels



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