Rhizobium anhuiense JSP2-5 and application thereof to improvement of tobacco-grown soil with crop rotation

A technology of rhizobia and bacterial fertilizer, applied in the field of microorganisms, to achieve the effect of improving soil microbial community structure, increasing the number, increasing yield and nitrogen, phosphorus and potassium content

Inactive Publication Date: 2018-05-15
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 anhuiense JSP2-5 and application thereof to improvement of tobacco-grown soil with crop rotation
  • Rhizobium anhuiense JSP2-5 and application thereof to improvement of tobacco-grown soil with crop rotation
  • Rhizobium anhuiense JSP2-5 and application thereof to improvement of tobacco-grown soil with crop rotation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation, purification and preservation of rhizobia JSP2-5

[0023] Collect the root nodules of wild strong red purple vetch in Jianzhu Township, Gulin, Luzhou, Sichuan Province, clean them, take off part of the root bark, absorb the surface moisture with absorbent paper, put them in anhydrous calcium chloride and cover them with absorbent cotton in the small tube. Carry out the following operations in the laboratory: the root nodules collected are soaked in sterile water for imbibition, soaked in 95% ethanol for 30s to eliminate surface tension, then sterilized with 0.1% (mass volume ratio) mercury chloride surface for 5min, and then use sterile Rinse with bacterial water 6 to 8 times, under the condition of aseptic operation, after clamping a single root nodule, 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 volume ratio) sodium molybdate 2mL, 1% (mass volume ra...

Embodiment 2

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

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

[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) Amplification of 16S rRNA gene and construction of phylogenetic tree

[0032] Using the total DNA as a t...

Embodiment 3

[0040] The making of embodiment 3 rhizobia inoculum

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

[0042] This rhizobia strain is inoculated in the liquid 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, ammonium molybdate (1%) 2mL, boric acid (1%) 2mL, water 1000mL), cultured in a shaker at 28°C for 2 days. Peat that has passed through a 100-mesh sieve is selected as the strain carrier, and the formula is: 488.5 g of peat, 1 g of sucrose, 0.5 g of superphosphate (passed through a 100-mesh sieve), 0.005 g of so...

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Abstract

The invention discloses Rhizobium anhuiense JSP2-5 and an application thereof to improvement of tobacco-grown soil with crop rotation. Rhizobium anhuiense JSP2-5 is preserved in a China Center for Type Culture Collection, and preservation number is CCTCC NO:M 2017618. A strain of Rhizobium anhuiense JSP2-5 adapted to plantation of Vicia villosa in Sichuan region is obtained, and Rhizobium anhuiense JSP2-5 can be used for substantially improving yields and contents of nitrogen, phosphor and potassium of Vicia villosa; Rhizobium anhuiense is turned over and a test shows that Rhizobium anhuiensefor inoculating Vicia villosa in the tobacco-grown soil with crop rotation can effectively increase contents of organic matters, rapidly available phosphorus, alkali-hydrolyzable nitrogen, rapidly available potassium, and quantity of three large groups of microorganisms in soil, improve microbial community structures of soil, and realize effects of soil fertility.

Description

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

Claims

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Application Information

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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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