普萨土壤杆菌的遗传毒性研究

doi:10.3969/j.issn.1004-616x.2025.04.018

摘要/Abstract

摘要： 目的：研究普萨土壤杆菌的遗传毒性，为其进一步的毒理学研究和在食品工业中的应用提供科学依据和数据支持。方法：细菌回复突变试验(Ames试验)采用TA97a、TA98、TA100、TA102和TA1535共5种菌种，设0.008、0.04、0.2、1、5 μL/皿共5个剂量组，另设蒸馏水对照组、空白对照组、二甲基亚砜对照组(DMSO)和阳性对照组，在加和不加S₉的试验条件下进行平板掺入法试验，每组作3个平行皿，重复试验1次；哺乳动物红细胞微核试验将昆明小鼠按雌雄分别随机分为5组，每组含雌雄鼠各5只，设2 500、5 000和10 000 mg/kg共3个剂量组，另设阴性对照组和阳性对照组(CP，40 mL/kg)；体外哺乳细胞染色体畸变试验采用中国仓鼠肺细胞(CHL)，±S₉组设阴性对照组(蒸馏水)、阳性对照组(+S₉，15 μg/mL CP；-S₉，0.20 μg/mL MMC)和1.25、2.5和5 μL/mL共3个剂量组。结果： Ames试验中，受试物对菌株无毒性，在±S₉条件下，受试物各剂量组的平均回变菌落数均小于阴性对照组的2倍，表明该受试物未诱导5种菌株回变菌落数增加；阳性对照组的平均回变菌落数均大于阴性对照组2倍。哺乳动物红细胞微核试验中，3个剂量组雌性昆明小鼠的含微核细胞率均值分别为2.0‰、2.1‰和2.4‰，与阴性对照组(均值1.8‰)相比较差异均无统计学意义(P>0.05)；3个剂量组雄性昆明小鼠的含微核细胞率均值分别为2.0‰、2.3‰和2.5‰，与阴性对照组(均值1.9‰)相比较差异均无统计学意义(P>0.05)；环磷酰胺阳性对照组显著升高(P<0.01)。体外哺乳类细胞染色体畸变试验中，在＋S₉条件下，3个剂量组的染色体总畸变率分别为2%、3%和6%，与阴性对照组(2%)比较差异均无统计学意义(P>0.05)，在-S₉条件下，3个剂量组的染色体总畸变率分别为2%、3%和5%，与阴性对照组的2%比较，差异均无统计学意义(P>0.05)，而阳性对照组则显著升高(P<0.01)。结论：在本试验条件下，普萨土壤杆菌无遗传毒性。

关键词: 普萨土壤杆菌, 细菌回复突变试验, 微核试验, 染色体畸变试验, 遗传毒性

Abstract: OBJECTIVE： To study the genotoxicity of Agrobacterium pusense in order to provide scientific basis and data support for further toxicological research and application in the field of food industry. METHODS： The bacterial reversion mutation test (Ames test) was conducted using TA97a，TA98，TA100，TA102 and TA1535 strains，with five dose groups of 0.008，0.04，0.2，1 and 5 μL/plate，solvent control group (distilled water)，untreated control group，dimethyl sulfoxide control group (DMSO) and positive control group were also set up. The standard plate incorporation method was used in the study in the presence or absence of S₉ metabolic activation system with 3 parallel plates in each group and repeated once. In mammalian erythrocyte micronucleus test，KM mice were randomly divided into 5 groups according to male and female，with 5 animals per sex in each group. Three dose groups were set up as 2 500，5 000 and 10 000 mg/kg，and negative control group and positive control group (CP，40 mL/kg) were also set up. Chinese hamster lung cells (CHL) were used in chromosome aberration test in vitro，and the chromosome aberration test was performed in 1.25，2.50 and 5.00 μg/mL dose groups，respectively，as well as in negative control group (distilled water) and positive control group (+S₉，15 μg/mL CP；-S₉，0.20 μg/mL MMC). The test was conducted in the presence or absence of S₉ metabolic activation system. RESULTS： In the Ames test，the test article was not toxic. In the presence or absence of S₉，the average number of recurrent colonies in each dose group was less than 2 times that of the negative control group，indicating that the tested article did not induce the increase of reverted colony number，and the average number of recurrent colonies in the positive control group was 2 times higher than that in negative control group. In the micronucleus test of mammalian erythrocytes，the average rates of micronucleated cells in female KM mice in the 3 dose groups were 2.0‰，2.1‰ and 2.4‰，respectively，which had no statistical significance compared with the negative control group (1.8‰) (P>0.05). The average rates of micronucleus containing cells in male KM mice in the 3 dose groups were 2.0‰，2.3‰ and 2.5‰，respectively，which had no statistical significance compared with the negative control group (1.9‰) (P>0.05). While the positive control group cyclophosphamide significantly increased (P<0.01)；In vitro chromosome aberration test of mammalian cells，the total chromosome aberration rates of the 3 dose groups were 2%，3% and 6% with S₉，respectively，which was no significantly different compared with the negative control group (2%) (P>0.05). The total chromosome aberration rates of the 3 dose groups were 2%，3% and 5% without S₉，respectively. Compared with negative control group (2%)，there was no statistical significance (P>0.05)，while the positive control group was significantly increased (P<0.01). CONCLUSION： In the conditions of this study，Agrobacterium pusense was not genotoxic.

Key words: Agrobacterium pusense, Ames test, micronuclear test, chromosomal aberration test, genotoxicity

中图分类号:

R394.6

夏祺悦, 豆捷雄, 刘科亮. 普萨土壤杆菌的遗传毒性研究[J]. 癌变·畸变·突变, 2025, 37(4): 330-335.

XIA Qiyue, DOU Jiexiong, LIU Keliang. Study on the genotoxicity of Agrobacterium pusense[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(4): 330-335.

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