Quantitative Detection of Methanotrophs in Soil by Novel pmoA -Targeted Real-Time PCR Assays

甲烷单加氧酶 甲烷利用细菌 细菌 α蛋白细菌 蛋白质细菌 土壤微生物学 生物 土壤水分 16S核糖体RNA 化学 微生物学 环境化学 生物化学 甲烷厌氧氧化 生态学 遗传学 催化作用
作者
Steffen Kolb,Claudia Knief,Stephan Stubner,Ralf Conrad
出处
期刊:Applied and Environmental Microbiology [American Society for Microbiology]
卷期号:69 (5): 2423-2429 被引量:449
标识
DOI:10.1128/aem.69.5.2423-2429.2003
摘要

ABSTRACT Methane oxidation in soils is mostly accomplished by methanotrophic bacteria. Little is known about the abundance of methanotrophs in soils, since quantification by cultivation and microscopic techniques is cumbersome. Comparison of 16S ribosomal DNA and pmoA (α subunit of the particulate methane monooxygenase) phylogenetic trees showed good correlation and revealed five distinct groups of methanotrophs within the α and γ subclasses of Proteobacteria : the Methylococcus group, the Methylobacter/Methylosarcina group, the Methylosinus group, the Methylocapsa group, and the forest clones group (a cluster of pmoA sequences retrieved from forest soils). We developed quantitative real-time PCR assays with SybrGreen for each of these five groups and for all methanotrophic bacteria by targeting the pmoA gene. Detection limits were between 10 1 and 10 2 target molecules per reaction for all assays. Real-time PCR analysis of soil samples spiked with cells of Methylococcus capsulatus , Methylomicrobium album , and Methylosinus trichosporium recovered almost all the added bacteria. Only the Methylosinus -specific assay recovered only 20% of added cells, possibly due to a lower lysis efficiency of type II methanotrophs. Analysis of the methanotrophic community structure in a flooded rice field soil showed (5.0 ± 1.4) × 10 6 pmoA molecules g −1 for all methanotrophs. The Methylosinus group was predominant (2.7 × 10 6 ± 1.1 × 10 6 target molecules g −1 ). In addition, bacteria of the Methylobacter/Methylosarcina group were abundant (2.0 × 10 6 ± 0.9 × 10 6 target molecules g of soil −1 ). On the other hand, pmoA affiliated with the forest clones and the Methylocapsa group was below the detection limit of 1.9 × 10 4 target molecules g of soil −1 . Our results showed that pmoA -targeted real-time PCR allowed fast and sensitive quantification of the five major groups of methanotrophs in soil. This approach will thus be useful for quantitative analysis of the community structure of methanotrophs in nature.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
2秒前
2秒前
zzzz发布了新的文献求助10
3秒前
小马完成签到,获得积分10
5秒前
5秒前
一朵云完成签到,获得积分10
6秒前
JSY发布了新的文献求助30
7秒前
浩铭完成签到,获得积分10
8秒前
Iven发布了新的文献求助10
8秒前
10秒前
12秒前
冷酷的天晴完成签到,获得积分10
12秒前
ysy完成签到,获得积分10
13秒前
13秒前
yum完成签到,获得积分20
14秒前
Ava应助专注的白柏采纳,获得10
14秒前
何如当初莫相识完成签到,获得积分10
14秒前
15秒前
落后从阳发布了新的文献求助10
17秒前
372925abc完成签到,获得积分10
17秒前
18秒前
清秀颜演完成签到,获得积分10
18秒前
carbonhan完成签到,获得积分10
19秒前
sh131完成签到,获得积分10
19秒前
复杂念梦完成签到 ,获得积分10
20秒前
DukeTao完成签到,获得积分10
20秒前
20秒前
bbb发布了新的文献求助10
20秒前
芝士椰果完成签到,获得积分10
21秒前
21秒前
华仔应助zy采纳,获得10
21秒前
飘逸问薇完成签到 ,获得积分10
23秒前
U2完成签到,获得积分10
24秒前
LIUYONG发布了新的文献求助10
24秒前
害羞的书芹完成签到,获得积分10
25秒前
rational完成签到,获得积分10
25秒前
支雨泽完成签到,获得积分10
25秒前
Clairezg完成签到 ,获得积分10
26秒前
量子星尘发布了新的文献求助10
26秒前
Passskd发布了新的文献求助10
26秒前
高分求助中
【提示信息,请勿应助】关于scihub 10000
Les Mantodea de Guyane: Insecta, Polyneoptera [The Mantids of French Guiana] 3000
徐淮辽南地区新元古代叠层石及生物地层 3000
The Mother of All Tableaux: Order, Equivalence, and Geometry in the Large-scale Structure of Optimality Theory 3000
Global Eyelash Assessment scale (GEA) 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 550
Research on Disturbance Rejection Control Algorithm for Aerial Operation Robots 500
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 4038368
求助须知:如何正确求助?哪些是违规求助? 3576068
关于积分的说明 11374313
捐赠科研通 3305780
什么是DOI,文献DOI怎么找? 1819322
邀请新用户注册赠送积分活动 892672
科研通“疑难数据库(出版商)”最低求助积分说明 815029