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Specific fragment for nitrate-assimilation bacillus megatherium and application thereof

A technology of Bacillus megaterium and nitrate, applied in the direction of DNA/RNA fragments, microbial determination/inspection, biochemical equipment and methods, etc., can solve problems such as high multiple cropping index, soil salinization, high fertilizer application rate, etc.

Inactive Publication Date: 2018-12-07
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of matching soil, water and fertilizer management measures for protected agriculture, coupled with the long-term high intensification, high multiple cropping index, and high fertilizer application rate of protected cultivation, soil salinization has been caused.

Method used

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  • Specific fragment for nitrate-assimilation bacillus megatherium and application thereof
  • Specific fragment for nitrate-assimilation bacillus megatherium and application thereof
  • Specific fragment for nitrate-assimilation bacillus megatherium and application thereof

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Embodiment Construction

[0027] In this embodiment, based on the above-mentioned specific fragment verification method, the Bacillus megaterium was detected in the soil, and the specific steps u include:

[0028] Primers were designed for the three fragments, and 3 pairs of primer fragments were obtained. Extraction facility soil (CK-S) total DNA, facility soil total DNA (CK-N) containing Bacillus megaterium of the present invention, Bacillus megaterium (N) of the present invention, one strain of Bacillus megaterium (C1), two strains of Bacillus subtilis Bacillus (C2, C3) and two strains of Bacillus colloidus (C4, C5) were amplified by PCR and reversed to verify that corresponding amplified bands were obtained, and three specific fragments were sequenced. The specific operation steps are as follows:

[0029] Step i) Extraction of bacterial genomic DNA: collect 1.0×109 (OD600 of 1mL bacterial solution is 1-1.5) bacterial culture in a 2mL centrifuge tube, centrifuge at 12,000×g for 30s, discard the sup...

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Abstract

The invention discloses a specific fragment for nitrate-assimilation bacillus megatherium and an application thereof. The fragments are specifically shown as Seq ID No. 1-No.3. The bacillus megatherium comprises assimilated nitrate reductase and nitrite reductase genes, is capable of improving secondary salinity soil by utilizing a biological enzyme method, and has the characteristics of being rapid in efficiency and environment-friendly. By performing whole genome sequencing on the nitrate-assimilation bacillus megatherium and performing GenBank database whole-genome sequence parallel comparison, the specific sequence fragments are finally obtained. By virtue of the specific fragments disclosed by the invention, the bacillus megatherium can be rapidly and accurately detected from the environment, and related engineering bacteria are prepared according to the specific fragments.

Description

technical field [0001] The invention relates to a technique in the field of bioengineering, in particular to a specific fragment of nitrate assimilating Bacillus megaterium and its application. Background technique [0002] Due to the development of facility agriculture and its important role in the off-season and cross-regional planting of vegetables and other important economic crops in my country, facility agriculture has been widely promoted and applied in my country. However, due to the lack of matching facility agriculture soil, water and fertilizer management measures, coupled with the long-term high intensification, high multiple cropping index, and high fertilizer application rate of facility cultivation, soil salinization has resulted. [0003] The accumulation of nitrate is one of the main characteristics of soil secondary salinization. Due to the application of a large amount of chemical fertilizers, especially nitrogen fertilizers, the problem of secondary salin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/689C12Q1/04
CPCC12Q1/689
Inventor 周培蒋淼张丹支月娥初少华陈寻峰
Owner SHANGHAI JIAO TONG UNIV
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