转cry1Ac基因欧洲黑杨对赤子爱胜蚓生长发育和生殖的影响

doi:10.11707/j.1001-7488.20211209

摘要/Abstract

摘要：

目的: 调查转cry1Ac基因欧洲黑杨(世纪杨)对土壤指示生物蚯蚓生长、繁殖和体内生物酶活性的影响，为转基因杨树大规模应用提供生态安全方面的数据。方法: 以非转基因杨树为对照，将转基因欧洲黑杨(世纪杨)叶片添加到土壤中连续喂养赤子爱胜蚓140天，每隔14天调查蚯蚓存活情况，测定蚯蚓体质量变化，统计蚯蚓繁殖产生蚓茧数量及蚓茧孵化情况，在蚓茧孵化7天后和成蚓暴露140天后测定蚯蚓体内总蛋白、乙酰胆碱酯酶(AchE)、谷胱甘肽过氧化物酶(GST)和超氧化物歧化酶(SOD)活性的变化。结果: 与对照杨树相比，饲喂转基因杨树叶片的蚯蚓存活率未发生显著改变；各处理组蚯蚓的体质量随试验进行而增加，140天10次调查中转基因杨树和对照杨树处理组蚯蚓体质量均无显著性差异；转基因杨树和对照杨树处理组均于试验第2次调查(28天)时开始繁殖，整个试验期转基因杨树和对照杨树处理组单条蚯蚓产蚓茧数无显著性差异，42~140天8次调查中，仅在第70天时对照杨树处理组蚓茧数显著高于转基因杨树组，其余7次调查均无显著差异；转基因杨树和对照杨树处理组蚓茧孵化率和每茧平均新生蚓数量无显著性差异；暴露140天，蚯蚓体内乙酰胆碱酯酶(AchE)、谷胱甘肽过氧化物酶(GST)和超氧化物歧化酶(SOD)活性在转基因杨树和对照杨树处理组无显著性差异；通过ELISA法未在蚯蚓组织检测到外源Cry1A蛋白，说明长时间饲喂(140天)转基因杨树叶片，外源蛋白不会在蚯蚓体内蓄积。结论: 相对对照杨树，饲喂转基因杨树叶片对赤子爱胜蚓的生存、生长和繁殖无不利影响。

关键词: 欧洲黑杨, 转cry1Ac基因, 赤子爱胜蚓, 生长, 繁殖, 体内酶活性

Abstract:

Objective: This study aimed to investigate the effect of the cry1Ac transgenic Populus nigra (Shijion) on the growth, reproduction and enzyme activity of the earthworm Eisenia foetida, a soil indicator organism. Method: In this study, the leaves of the cry1Ac transgenic and non-transgenic poplar were added to the soil for continuously feeding to the earthworm E. foetida for 140 days in the laboratory. The survival of earthworm, the change of earthworm weight, and the number of the cocoons and their hatching were investigated every 14 days. The total protein, and the activities of acetylcholinesterase (Ache), glutathione peroxidase (GST), and superoxide dismutase (SOD) in earthworm were determined after 7 days of cocoon incubation and 140 days of adult exposure. Cry1Ac concentrations in the soil, casts and guts of earthworm were also detected by ELISA. Result: Over the 140 d experimental period, there were no significant differences in survival rate, and weight of earthworms fed with the transgenic and non-transgenic poplar leaves. Both the transgenic and non-transgenic poplars treatments began to breed at the 28^th day. There was no significant difference in the number of cocoons produced by a single earthworm between the transgenic poplar and non-transgenic poplar treatments in the whole experimental period. In the 8 investigations of 42-140 days, the average number of cocoons in non-transgenic poplar treatment was significantly higher than that in the transgenic poplar treatment only at the 70^th d, and there was no significant difference at the other 7 investigation periods. There was no significant difference in the hatching rate of earthworm cocoons and the average number of newborn earthworms per cocoon between transgenic poplar and control poplar treatments. There were no significant differences in the total protein content, and the activities of AchE, GST, and SOD in adult E. foetida after 140 days of exposure between transgenic and non-transgenic poplar treatments. On the 7^th d, SOD activity of new worm increased under two poplar treatments, compared with the fermented cow dung treatment. ELISA determined a sharp decrease of Cry1Ac in mixed soil along with the experimental time. No Cry1Ac was detected in the earthworm tissue under long-term feeding. Conclusion: The results suggests that transgenic poplar does not have significant adverse effects on the survival, growth and reproduction of E. foetida.

Key words: Populus nigra, transgenic cry1Ac, Eisenia foetida, growth, reproduction, enzyme activity

中图分类号:

S792.11

沈文静,张莉,刘来盘,方志翔,刘标. 转cry1Ac基因欧洲黑杨对赤子爱胜蚓生长发育和生殖的影响[J]. 林业科学, 2021, 57(12): 92-98.

Wenjing Shen,Li Zhang,Laipan Liu,Zhixiang Fang,Biao Liu. Effects of cry1Ac Transgenic Populus nigra on Growth, Development, and Reproduction of the Earthworm Eisenia foetida[J]. Scientia Silvae Sinicae, 2021, 57(12): 92-98.

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指标 Characteristics	非转基因杨树 Non-transgenic poplar	转基因杨树 Transgenic poplar
总蛋白 Total protein	103.76±18.2 a	117.15±21.85
总糖 Total sugar	35.67±6.06 a	32.39±7.39 a
总氮 Total N	18.17±3.57 a	16.54±1.67 a
总磷 Total P	3.48±0.45 a	3.14±0.14 a
Cry1Ac蛋白 Cry1Ac protein	ND	(1.42±0.13)×10^-3

处理Treatments	时间Time/d
处理Treatments	14	28	42	56	70	84	98	112	126	140
发酵牛粪 Fermented cow dung	100	100	100	100	100	93.33± 8.73	93.33± 8.73	90.00± 7.56	86.67± 4.36	83.33± 4.36
非转基因杨树 Non-transgenic poplar	100	100	96.39± 4.36	96.39± 4.36	96.39± 4.36	93.33± 8.73	93.33± 8.73	90.00± 7.566	90.00± 7.56	86.67± 11.55
转基因杨树 Transgenic poplar	100	100	100	100	96.39± 4.36	96.39± 4.36	96.39± 4.36	93.33± 8.76	90.00± 7.56	86.67± 4.36

处理 Treatments	蚓茧孵化率 Hatchability(%)	新生蚓(条/茧) Number of new earthworms
发酵牛粪 Fermented cow dung	73.33±4.36 a	2.36±0.97 a
非转基因杨树 Non-transgenic poplar	70.00±15.12 a	2.23±1.10 a
转基因杨树 Transgenic poplar	66.67±8.73 a	2.25±0.93 a

发育阶段 Developmental stage	处理 Treatments	Total protein/ (mg·g^-1)	SOD/ (μmol·g^-1min^-1)	GST/ (μmol·g^-1min^-1)	AChE/ (μmol·g^-1min^-1)
成蚓 Adult earthworm (140 d)	发酵牛粪Fermented cow dung	22.66±3.30 a	180.07±37.39 a	69.11±2.53 a	6.26±1.76 a
	非转基因杨树Non-transgenic poplar	19.31±1.29 a	193.17±54.34 a	69.84±12.45 a	6.08±1.04 a
	转基因杨树Transgenic poplar	19.53±1.69 a	174.52±33.70 a	67.74±6.59 a	7.34±1.31 a
新生幼蚓 new earthworm(7 d)	发酵牛粪Fermented cow dung	19.68±3.07 a	169.74±24.68 a	70.62±5.23 a	5.69±0.66 a
	非转基因杨树Non-transgenic poplar	22.35±2.56 a	294.65±42.81 b	78.20±11.20 a	5.92±0.62 a
	转基因杨树Transgenic poplar	21.37±1.74 a	276.20±31.45 b	81.31±12.72 a	6.14±0.79 a

样品 Samples	时间Time/d
样品 Samples	0	14	84	140
叶片土壤混合物Leaf and soil mixture	38.48±7.93	23.84±4.32	16.67±2.17	13.83±3.00
蚯蚓组织Whole earthworm tissue	ND	NT	NT	ND
蚯蚓排泄物Earthworm excrement	ND	NT	NT	4.07±0.79