培养基组成及微生物种类归属对白磷生物破坏的影响

A. Mindubaev, E. Babynin, E. Badeeva, S. Minzanova, L. Mironova, A. Arbuzov
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引用次数: 0

摘要

本研究比较了黑曲霉菌株AM1在含P4为唯一磷源的不同组成培养基中的生长情况。在10种培养基中,选择了曲霉生长最快的两种培养基。这些培养基被认为是最适合生长的。通过比较培养基的组成和曲霉在其中的生长速度,我们发现了一个关键成分,它是AM1生长和白磷生物降解的有利因素。该成分为硝酸钠(NaNO3)。研究还表明,硫酸铜(CuSO4)对含白磷培养基中曲霉的生长没有影响,无论这些培养基的成分如何。这个结果与我们以前的发现是一致的。此外,在本工作中,首次描述了将培养基中白磷浓度提高到1%以上的尝试。为此,我们在培养基中加入了以下溶剂:二甲基亚砜(DMSO)和柴油,其中白磷溶解较好。显然,这些物质的存在会对Aspergill的生长产生不利影响。因此,进一步增加P4浓度的问题仍然是一个没有答案的问题。白磷即使在室温下也能与二价铜离子发生反应。而我们选择的普里达姆-戈特利布介质,其成分中含有硫酸铜。白磷乳剂的加入导致了黑色沉淀物的形成,这是化学反应发生的证据。因此,微生物是在白磷的化学转化产物中生长的,而不是在白磷的存在下生长的,而且实验也不是完全干净的。因此,在本研究中,我们对Pridham-Gottlieb营养培养基进行了进一步的改性,不仅排除了作为磷源的磷酸盐,还排除了硫酸铜。此外,我们还将我们的黑曲霉菌株AM1和AM2与来自全俄罗斯微生物收集(ARCM)的3株菌株(菌株FW-650, *通讯作者E-mail地址:mindubaev-az@yandex.ru (A.Z. Mindubaev) Biogeosystem Technique, 2019, 6(2) 103 FW-2664和FW-2731)以及4种不同细菌进行了白磷抗性比较。虽然菌株AM1的抗性最高,但来自ARCM的3株黑曲霉对白磷的抗性也高于菌株AM1。结果表明,在含白磷的营养培养基中除去硫酸铜并不会阻止真菌的生长。此外,白磷不会与沉淀物的形成发生反应,在这些条件下停留的时间更长。这一事实是支持生物降解的重要论据,并在白磷的微生物解毒方法中具有实际适用性。然而,与ARCM菌株相比,AM1菌株的抗性仅在含铜培养基中观察到。显然,菌株AM1对白磷与Cu2+反应产生的有毒产物的抗性最强。我们之前的研究首次证明了白磷中存在遗传毒性。毫无疑问,这使得白磷处理起来更加危险。然而,我们最初的研究是在原核生物(鼠伤寒沙门氏菌)上进行的。由于原核生物的遗传装置与真核生物(包括人类)的排列方式不同,沙门氏菌的研究结果不能完全转移到人类身上。除了Ames试验和SOS-lux试验所研究的基因突变(它们在所有生物体中都具有共同的性质)之外,真核生物中还应该研究基因组重排。为此,对洋葱的根(Allium cepa L.)进行了葱试验。本文首次用葱试验法研究了白磷对真核生物细胞周期的负面影响。结果表明,即使白磷浓度很低,只有0.01%,也会成倍增加染色体畸变的数量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The Effect of Culture Media Composition and Microorganism Species Affiliation on the Biological Destruction of White Phosphorus
The presented study compared the growth of Aspergillus niger strain AM1 in culture media varying in composition but containing P4 as the sole source of phosphorus. Of the ten media, two in which Aspergillus grew the fastest were selected. These media were concluded to be optimal for growth. Comparing the compositions of the media and the growth rate of Aspergillus in them, we found a key component that is a favorable factor for the growth of AM1 and the biodegradation of white phosphorus. This component was sodium nitrate (NaNO3). It has also been shown that copper sulphate (CuSO4) has no effect on the growth of Aspergillus in media with white phosphorus, regardless of the composition of these media. This result is in harmony with our previous findings. Furthermore, in the present work, attempts to increase the concentration of white phosphorus in the culture medium to values above 1 % are described for the first time. For this purpose, we added the following solvents to the culture media: dimethyl sulfoxide (DMSO) and diesel, in which white phosphorus dissolves relatively well. Apparently, the presence of these substances adversely affects the growth of Aspergill. Therefore, the problem of further increasing the concentration of P4 remains an unanswered. White phosphorus, reacts with ions of divalent copper even at room temperature. and the Pridham-Gottlieb medium, which we have chosen for our purposes, contains copper sulfate in its composition. The addition of an emulsion of white phosphorus led to the formation of a black precipitate, which is evidence that a chemical reaction took place. Thus, the growth of microorganisms occurred in the presence of not so much white phosphorus as the products of its chemical transformations, and the experiments were not completely clean. Therefore, in the present study, we carried out further modification of the Pridham-Gottlieb nutrient medium, excluding from it not only phosphates as a source of phosphorus, but also copper sulfate. In addition, we compared the white phosphorus resistance of our A. niger strains AM1 and AM2, with three strains from the All-Russian Collection of Microorganisms (ARCM) (strains FW-650, * Corresponding author E-mail addresses: mindubaev-az@yandex.ru (A.Z. Mindubaev) Biogeosystem Technique, 2019, 6(2) 103 FW-2664 and FW-2731), as well as four different bacterial species. Though highest resistance was observed in strain AM1, the three strains of A. niger, sent from ARCM, also showed a higher resistance to white phosphorus than the bacteria. It was shown that exclusion of copper sulfate from the composition of the nutrient medium with white phosphorus does not prevent the growth of fungi. In addition, white phosphorus does not react with the formation of a precipitate and remains for a longer period under these conditions. This fact is a serious argument in favor of biodegradation and has practical applicability in the method of microbial detoxification of white phosphorus. However, a higher resistance of AM1 in comparison to the ARCM strains was only observed in a medium with copper. Apparently, strain AM1 is most resistant to the toxic products from the reaction of white phosphorus with Cu2+. Our previous studies demonstrated for the first time the presence of genotoxic properties in white phosphorus. This in no doubt makes white phosphorus even more dangerous to handle. However, our initial studies were carried out on prokaryotes (Salmonella typhimurium). Since the genetic apparatus of prokaryotes is differently arranged than in eukaryotes (including humans), the results of the studies on Salmonella is not completely transferable to humans. In addition to the gene mutations studied by the Ames test and the SOS-lux test, which have a common nature in all living organisms, there are genomic rearrangements that should be studied in eukaryotes. For this purpose, an Allium test is used on onion rootlets (Allium cepa L.). In this work, we present the first report on the negative effect of white phosphorus on the cell cycle of eukaryotes by the Allium test method. It turned out that white phosphorus, even at very low concentrations of 0.01 %, exponentially increases the number of chromosomal aberrations.
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