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A kind of rhizobia axlq16 and its application

A technology of rhizobia and bacterial strains, applied in the field of microorganisms, to achieve the effects of enriching group diversity, high nodulation rate, and strong nitrogen fixation ability

Inactive Publication Date: 2020-06-16
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of mature and effective rhizobia agents for high-efficiency nitrogen-fixing peanuts

Method used

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  • A kind of rhizobia axlq16 and its application
  • A kind of rhizobia axlq16 and its application
  • A kind of rhizobia axlq16 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation and Purification of Rhizobium AXLQ16

[0023] (1) Take fresh, mature and large nodules of peanuts, rinse them with sterilized water, and dry the surface water with filter paper.

[0024] (2) Put it in 95% alcohol for 3-5 minutes, then add 1g / L HgCl 2 medium, sterilize for 3 to 5 minutes, rinse with sterile water 5 to 6 times after taking it out.

[0025] (3) Cut in half on the flame-sterilized glass slide, hold the half tumor with sterile tweezers, and make the incision face the surface of the YMA medium to draw a line. The formula of the YMA medium is shown in Table 1. Cultivate at ℃.

[0026] (4) After the bacteria grow, use a sterilized toothpick to scrape a small amount of rhizobia colonies from the plate, dilute them with sterile water, and culture them on the plate again. Observe the colonies after 3 days until 15 days (bradyrhizobia need 7 to 15 days to appear colonies). If no monoclonal colonies appear, it is necessary to repeatedly strea...

Embodiment 2

[0033] Example 2 of Rhizobium AXLQ16 16s rDNA sequence sequencing

[0034] Perform PCR specific amplification on rhizobia monoclonal liquid, primers are V3-V4-F and V4-V5-R respectively, forward primer V3-V4-F is 5′-GWATTACCGCGGCKGCTG-3′; reverse primer V4 -V5-R is 5′-CCGTCAATTCMTTTRAGTTT-3′, and the enzyme is 2×star Mix. The PCR amplification products were detected by electrophoresis imaging technology to observe whether they had bands, and the remaining PCR amplification products were used for sequence determination. The sequencing result is shown in SEQ ID NO:1. The PCR reaction system is shown in Table 2:

[0035] Table 2 16s rDNA 2×starMix enzyme reaction system

[0036]

[0037] The sequences of 12 reference strains were obtained from the NCBI (GenBank) database, and the sequences of the isolated strains and reference strains were analyzed using the software BioEdit and MEGA6. 16s The rDNA partial sequence was analyzed, and the phylogenetic tree of the isolate...

Embodiment 3

[0038] Example 3 Hydroponics back inoculation test

[0039] 1. Strain cultivation: transfer the tested strains stored at -80°C to YMA liquid medium for liquid culture, and cultivate to the logarithmic phase. The growth of the strains is detected by a nucleic acid and protein analyzer, and the OD value is used to calculate the bacterial content. The formula of YMA (YeastMannnitol Agar) liquid medium is: weigh 10 g of mannitol, MgSO 4 ∙7H 2 O 0.2g, NaCl 0.1g, yeast powder 3g, K 2 HPO4 0.25 g, KH 2 PO 4 0.25 g, CaCO 3 3 g (added during storage), 15 g of agar was dissolved in 1 L of pure water.

[0040] 2. Sterilization treatment: Wrap the 2.5L plastic bucket, beaker, tweezers, Petri dish, filter paper, and glass rod required for the test, pack the vermiculite in a fresh-keeping bag, and sterilize at 121°C for 30 minutes. Pack YMA liquid medium and distilled water in bottles, and sterilize at 121°C for 20 minutes.

[0041] Put peanut seeds in 1g / L HgCl 2 Sterilize the ...

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Abstract

The invention discloses a kind of rhizobia AXLQ16 and its application. The strain is obtained from the rhizobia in the soil of Anxi area, Fujian Province by Minhua No. 11 peanut variety, the rhizobia is separated and purified from fresh nodules, and the The 16S rDNA partial sequence was determined, and through phylogenetic analysis, it was identified as the genus Rapid-growing Rhizobium ( Rhizobium ) of the new strain and named it Funong No. 1. The rhizobia AXLQ16 of the present invention has been proved to have the characteristics of high-efficiency nodulation and strong nitrogen fixation ability through laboratory hydroponic backgrafting tests and field backgrafting experiments. Inoculation of the rhizobia AXLQ16 can increase peanut biomass and yield, and can be applied to acidic soil in South China , as one of the measures for conventional cultivation of peanuts.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a rhizobia AXLQ16 and an application thereof. Background technique [0002] Rhizobium ( rhizobium ) is a kind of Gram-negative bacteria widely distributed in soil, which can form root nodule or stem nodule after infecting the roots or stems of legumes, and fix N in the air in the form of symbionts. 2 Become NH that can be absorbed by plants 4 + . The symbiotic system between legumes and rhizobia plays an extremely important role in the whole biological nitrogen fixation. It is estimated that about 100 million tons of nitrogen are fixed from the air on the surface of the earth every year, and about 55 million tons of nitrogen are fixed by leguminous plants and rhizobia. Rhizobium and leguminous plants form a symbiotic nitrogen fixation system. In addition to meeting the nitrogen needs of the plants themselves, they can also provide excess nitrogen to non-legumes tha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/20A01P21/00C12R1/41
CPCA01N63/00C12N1/205C12R2001/41
Inventor 廖红刘鹏钟永嘉田颖哲
Owner FUJIAN AGRI & FORESTRY UNIV
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