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Rhizobium jsp2-5 and its application in improving tobacco soil rotation

A rhizobia and soil technology, applied in the field of microorganisms, to increase the number, increase yield and nitrogen, phosphorus and potassium content, and improve the effect of soil microbial community structure

Inactive Publication Date: 2020-06-30
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the inoculation of rhizobia in leguminous green manure. To improve the symbiotic nitrogen fixation of rhizobia, the strain and the species of legumes are two fundamental factors.

Method used

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  • Rhizobium jsp2-5 and its application in improving tobacco soil rotation
  • Rhizobium jsp2-5 and its application in improving tobacco soil rotation
  • Rhizobium jsp2-5 and its application in improving tobacco soil rotation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation, purification and storage of Rhizobium JSP2-5

[0023] Collect wild and robust red vetch root nodules in Gulinjianzhu Township, Luzhou, Sichuan Province, clean them, remove part of the root bark, absorb the surface water with absorbent paper, and install them in anhydrous calcium chloride covered with absorbent cotton In the small tube. Perform the following operations in the laboratory: soak the collected nodules in sterile water for swelling, soak them in 95% ethanol for 30s to eliminate surface tension, then disinfect the surface with 0.1% (mass to volume) mercury for 5 minutes, and then use no Rinse with bacteria water for 6-8 times. In the case of aseptic operation, clip a single nodule and add Congo Red YMA medium (mannitol 10g, yeast powder 0.8g, KH 2 PO 4 0.25g, MgSO 4 .7H 2 O 0.2g, CaCl 2 .6H 2 O 0.1g, NaCl 0.1g, 1% (mass to volume ratio) sodium molybdate 2mL, 1% (mass to volume ratio) boric acid 2mL, 1% (mass to volume ratio) Congo red 2.5mL, ...

Embodiment 2

[0026] Example 2 Amplification and phylogenetic analysis of the 16S rRNA gene and other housekeeping genes glnII, atpD, recA of Rhizobium vulgaris JSP2-5

[0027] The total DNA of the strain was extracted, and the above 4 genes were amplified by PCR with the primers shown in Table 1. Bio-RAD MyCycler was used for the PCR reaction TM After the instrument, the PCR amplified products are detected on 1.0% agarose gel electrophoresis, they are sent to Chengdu Kinke Zixi Biotechnology Co., Ltd. for sequence determination. The gene amplification primers used in this study are listed in Table 1. The software DNAman 6.0 is used to calculate the similarity of gene sequence.

[0028] Table 1 PCR primers used in this experiment

[0029]

[0030] Note: Y = C or T, H = A, C or T, R = A or G, S = C or G, K = G or T, N = A, C, G or T, I = inosine, M = Aor C,N=any base.

[0031] (1) 16S rRNA gene amplification and phylogenetic tree construction

[0032] Using total DNA as a template, 16S rRNA was a...

Embodiment 3

[0040] Example 3 Preparation of rhizobia inocula

[0041] The strain JSP2-5 was isolated from the wild nodules of Vetch vetch in Gulin Jianzhu Township, Luzhou. Because of its strong nitrogen fixation ability, it was applied to the growth of Vetch vetch. The strain was identified as Rhizobium anhuiense, which was deposited in the Chinese Type Culture Collection in Wuhan University, Wuhan City, Hubei Province, China on October 23, 2017, with the deposit number: CCTCC NO:M 2017618.

[0042] The root nodule strain was inoculated into liquid YMA medium (10g mannitol, 0.8g yeast powder, KH 2 PO 4 0.25g, MgSO 4 .7H 2 O 0.2g, CaCl 2 .6H 2 O 0.1g, NaCl 0.1g, ammonium molybdate (1%) 2mL, boric acid (1%) 2mL, water 1000mL), shake culture in a shaker at 28°C for 2 days. Peat that has passed a 100-mesh sieve is selected as the strain carrier, and the formula: peat 488.5g, sucrose 1g, superphosphate (passed 100-mesh sieve) 0.5g, sodium molybdate 0.005g, and boric acid 0.005g. Adjust the pH to...

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Abstract

The invention discloses Rhizobium anhuiense JSP2‑5 and its application in improving tobacco crop rotation soil. Rhizobium anhuiense JSP2‑5 is preserved in China Center for Typical Culture Collection, and the preservation number is: CCTCC NO: M 2017618. The present invention has obtained a strain of rhizobia JSP2‑5 adapted to the cultivation of purple sweet potato in Sichuan area, and the rhizobia JSP2‑5 can significantly improve the yield and nitrogen, phosphorus and potassium content of purple sweet potato. Tobacco soil rotation inoculated with rhizobia purple vetch can effectively increase the content of soil organic matter, available phosphorus, alkaline nitrogen, and available potassium, and can increase the number of three major groups of microorganisms in the soil, improve the soil microbial community structure, and achieve fertilization. soil effect.

Description

Technical field [0001] The invention belongs to the field of microorganisms, and specifically relates to Rhizobium JSP2-5 and its application on improving tobacco soil rotation. Background technique [0002] Tobacco, as a special economic crop, must create a good soil environment for tobacco growth in order to achieve high quality and suitable yield of tobacco. With the rapid development of flue-cured tobacco production in China, long-term large-scale application of chemical fertilizers and unreasonable crop rotation have caused problems such as decline in tobacco yield and quality, soil compaction, decline in soil fertility, and reduced organic matter content, which have become the bottleneck for the sustainable development of tobacco production. In this context, the search for a cultivation method that can increase the yield and quality of tobacco leaves, improve the soil, and realize the sustainable development of tobacco production has become one of the urgent problems in flu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05G3/00C12R1/41
CPCC05F11/02C05G3/00C12N1/205C12R2001/41
Inventor 徐开未陈远学刘明龙玲陈强邹文豪余秀梅刘汉军
Owner SICHUAN AGRI UNIV
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