产硫化氢细菌的筛选及阻控苜蓿吸收铅和改良土壤的作用

doi:10.11686/cyxb2020265

摘要/Abstract

摘要：

为了探究产硫化氢细菌阻控牧草吸收重金属、修复重金属污染土壤的作用，从生长在矿区污染地的苜蓿中分离纯化产硫化氢细菌，根据菌株生理特性、产硫化氢能力、铅镉抗性以及吸附铅能力筛选高效菌株，然后以某农田受污染土壤为供试土壤，采用田间小区试验，研究高效菌株对紫花苜蓿生长及阻控苜蓿吸收Pb的效果与机制。结果表明，从生长在矿区污染地的苜蓿中分离筛选到一株高产硫化氢细菌Sar15，其具有合成吲哚乙酸（IAA）、铁载体和脲酶的特性，在溶液条件下能显著降低Pb²⁺的浓度（60%）。经16S rDNA分析将菌株Sar15归属于彭氏变形菌。菌株Sar15能够在紫花苜蓿根部定殖。与不接菌对照相比，菌株Sar15能显著增加受污染农田中生长的苜蓿地上部和根部干重、显著降低其Pb含量。菌株Sar15使得苜蓿地上部和根部Pb含量分别降低43%和45%。并且该菌株能够提高紫花苜蓿根际土壤pH和酶活性，降低根际土壤有效态Pb含量，减轻Pb对紫花苜蓿的毒害。Sar15处理具有修复中度Pb污染土壤、保障牧草安全生产的潜力。

关键词: 紫花苜蓿, Pb污染, 产硫化氢细菌

Abstract:

This research explored the roles of hydrogen sulfide-producing bacteria in preventing forages from absorbing heavy metals and in remediation of soil contaminated by heavy metals. From alfalfa growing at a locality contaminated by nearby mining activities， a bacterial strain， Sar15， with high hydrogen sulfide production was isolated. Sar15 was found to synthesize indole-3-acetic acid （IAA）， siderophores and urease， and to reduce the concentration of Pb²⁺in solution by 60% （P<0.05）. Using 16S rDNA analysis the strain Sar15 was identified as Proteus penneri. A field experiment was then conducted to evaluate the potential influence of this hydrogen sulfide-producing bacterial strain on the growth ofalfalfaand its role in the inhibition of Pb uptake by alfalfa in Pb-contaminated farmland. It was found that P. penneri strain Sar15 was the most efficient in producing hydrogen sulfide and could colonize on the roots of alfalfa. Compared with the control， the strain Sar15 increased the dry weight of alfalfa and significantly reduced its Pb content by 44%-45% （P<0.001）. P. penneri strain Sar15 also increased the rhizosphere soil pH and enzyme activity， reduced the available Pb content in the rhizosphere soil， and reduced the toxicity of Pb to alfalfa. Inoculationof P. penneri strain Sar15 has the potential to mitigate moderately Pb-contaminated soil and ensure safe forage production.

Key words: Medicago sativa, Pb pollution, hydrogen sulfide-producing bacteria

周倩倩, 张亚见, 张静, 殷涂童, 盛下放, 何琳燕. 产硫化氢细菌的筛选及阻控苜蓿吸收铅和改良土壤的作用[J]. 草业学报, 2021, 30(7): 44-52.

Qian-qian ZHOU, Ya-jian ZHANG, Jing ZHANG, Tu-tong YIN, Xia-fang SHENG, Lin-yan HE. Isolation of a beneficial hydrogen sulfide-producing bacterial strain that reduces lead uptake by Medicago sativa and aids remediation of Pb-contaminated soil[J]. Acta Prataculturae Sinica, 2021, 30(7): 44-52.

图/表 7

图1 菌株产H2S特性

Fig.1 Hydrogen sulfide-producing characteristics of strains

图2 菌株对溶液中铅浓度的影响*表示与CK相比，在0.05水平有显著差异(P<0.05)，**表示与CK相比，在0.01水平有极显著差异(P<0.01)，***表示与CK相比，在0.001水平有极显著差异(P<0.001)。下同。* indicates that there is a significant difference at the 0.05 level (P<0.05) compared to CK, ** indicates that there is a very significant difference at the 0.01 level (P<0.01), *** indicates a very significant difference at 0.001 level compared to CK (P<0.001). The same below.

Fig.2 Effect of strains on the concentration of Pb in solution

表1 菌株的生理特性

Table 1 Physiological characteristics of the isolated strains

菌株Strains	吲哚乙酸IAA （mg·L^-1）	铁载体Siderophore	脲酶Urease	Pb耐性Pb tolerance (mg·L^-1）	Cd耐性Cd tolerance (mg·L^-1）
Mr2	65.9	++	-	300	20
Mr3	4.1	+++++	+	500	20
Mr11	23.8	+++	-	500	10
Mr40	38.7	+++++	+	400	30
Sar15	15.8	++	+	800	50

表2 Sar15对紫花苜蓿干重、Pb含量、总量、转移系数和富集系数的影响

Table 2 Effects of strain Sar15 on the dry weight, Pb content, total content, translocation factor and bioconcentration factor of M. sativa

处理 Treatments	干重 Dry weight （g·5 plant^-1）		Pb含量 Pb content （mg·kg^-1）		Pb 总量 Total Pb content （μg）		转移系数 Translocation factor	根富集系数 Bioconcentration factor
处理 Treatments	地上部Shoot	根部Root	地上部Shoot	根部Root	地上部Shoot	根部Root	转移系数 Translocation factor	根富集系数 Bioconcentration factor
对照Control	1.2±0.10	0.4±0.05	8.7±0.70	13.0±0.10	10.7±0.20	5.2±0.90	0.66	0.033
活菌Sar15 Live Sar15	1.5±0.03*	0.5±0.08**	4.9±0.02***	7.2±0.10***	7.3±0.30***	3.6±0.30***	0.68	0.018***
灭活菌Sar15 Inactivation Sar15	1.3±0.03	0.4±0.03	8.4±0.80	12.0±0.30	10.5±0.90	5.3±0.50	0.70	0.032

图3 菌株Sar15对苜蓿根际土壤pH、DTPA提取态铅含量、脲酶和硫酸酯酶活性的影响

Fig.3 Effects of strain Sar15 on pH, DTPA-extractable Pb content, urease and sulfatase activity in rhizosphere soil of M. sativa

图4 菌株Sar15对苜蓿根际土壤酸可提取态Pb、可还原态Pb和可氧化态Pb含量的影响

Fig.4 Effects of strain Sar15 on the content of acid extractable Pb, reducible Pb and oxidizable Pb in M. sativa rhizosphere soil

图5 菌株Sar15在苜蓿根部的定殖

Fig.5 Colonization of strain Sar15 in the roots of M. sativa

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