Method for detecting plant root endophyte containing autotrophic arsenic oxidation function microorganisms

A technology of functional microorganisms and endophytes, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve problems such as unclear mechanism of heavy metal tolerance

Pending Publication Date: 2022-01-14
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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Problems solved by technology

[0004] Since the vast majority of plant endophytes cannot be obtained by pure culture techniques, although the community composition of plant endophytes has been discovered through molecular biology techniques, the heavy metal tolerance mechanism of plant root endophytes is still unclear, and further research is needed. Ecological interaction effects between functional endophytes and heavy metal polluted soil-plants

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  • Method for detecting plant root endophyte containing autotrophic arsenic oxidation function microorganisms
  • Method for detecting plant root endophyte containing autotrophic arsenic oxidation function microorganisms
  • Method for detecting plant root endophyte containing autotrophic arsenic oxidation function microorganisms

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Embodiment

[0048] A method for detecting whether plant root endophytes contain autotrophic arsenic oxidation functional microorganisms, comprising the following steps:

[0049] 1. Isolation of endophytic bacteria from plant roots

[0050] 1. Pretreatment of plant roots:

[0051] (1) Gently shake off the soil attached to the root of a centipede grass that has been polluted by arsenic in a mining area in Hechi, Guangxi for a long time, and then transfer the root system to a place containing 10mL MgSO 4 (10mM) solution in a centrifuge tube, vortexed for 5min, repeatedly removed the turbid liquid, and then transferred to 10mM MgSO containing Tween 20 (0.01%, V / V) 4 Vortex in the solution for 5 minutes, ultrasonic treatment for 15 minutes; while retaining the last buffer coating, it can be observed that there are more microorganisms growing ( figure 1 a);

[0052] (2) According to the above operation, add 10mL MgSO again 4 (10mM) solution was vortexed for 5min, transferred to 1% NaClO sol...

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Abstract

The invention discloses a method for detecting plant root endophyte containing autotrophic arsenic oxidation function microorganisms. Through a comprehensive means of combining separation, culture and DNA-SIP technologies, the functional microorganisms participating in the heavy (similar) metal oxidation-reduction process in the plant endophyte can be accurately distinguished. On the basis of efficiently extracting and separating plant tissue endophyte, endophyte liquid obtained through endophyte liquid separation is subjected to enlarged culture by using a liquid culture medium, so that the microorganism abundance and activity of the endophyte liquid are ensured. Meanwhile, DNA-SIP is used as a means, microorganisms for driving heavy metal redox can be directly and effectively anchored, q-PCR amplification and 16S rRNA high-throughput sequencing are further utilized, and the heavy metal redox function microorganism class group abundance change in a microcosm system and metabolism information of key function endophyte microorganisms are analyzed, and whether the plant endophyte community has the functional bacteria for driving the heavy (similar) metal redox process or not can be accurately judged.

Description

technical field [0001] The invention belongs to the field of microbial ecology, and in particular relates to a method for detecting microorganisms containing autotrophic arsenic oxidation function in plant root endophytes. Background technique [0002] Plant endophytes mainly live in plants and can exert various biological effects on host plants. Studies have shown that plant endophytes can enhance the resistance of host plants to abiotic stresses such as temperature, water, and metal pollution, as well as biotic stresses such as insects, pathogenic bacteria, and nematodes, and at the same time provide plants with the nutrients they need, such as nitrogen sources. Promote plant growth and regulate plant physiological processes. [0003] In recent years, plant endophytes have received widespread attention at home and abroad due to their unique ecology. A large number of studies have shown that endophytes participate in the physiological functions and various metabolisms of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/689C12Q1/6895
CPCC12Q1/04C12Q1/045C12Q1/689C12Q1/6895C12Q2600/158C12Q2600/178
Inventor 孙蔚旻孙晓旭裘浪黄端仪徐锐刘华清李永斌高文龙颜耕徐芙清
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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