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Rhizobium YZM0144 and application thereof

A rhizobia, M2018217 technology, applied in rhizobia YZM0144 and its application fields, to achieve the effects of increasing dry weight, strong nitrogen fixation ability, and increasing total nitrogen content

Inactive Publication Date: 2019-02-19
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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  • Rhizobium YZM0144 and application thereof
  • Rhizobium YZM0144 and application thereof
  • Rhizobium YZM0144 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation and purification of rhizobia YZM0144

[0023] (1) During the flowering period of normal peanut growth, take fresh whole peanut roots, take fresh, mature and large nodules of peanuts from the roots, rinse them with water, and dry the surface water with filter paper.

[0024] (2) Put it into ethanol with a volume fraction of 95% for 3 to 5 minutes, take it out and wash it with sterile water for 5 to 6 times, and then put it in 1g / L HgCl 2 Sterilize for 3-5 minutes, take it out and rinse with sterile water 5-6 times.

[0025] (3) Cut in half on a flame-sterilized glass slide, hold half of the tumor with sterile tweezers, make the incision face the surface of YMA (Table 1) culture medium, and incubate at 28°C after inversion.

[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 ...

Embodiment 2

[0033] Example 2 of Rhizobium YZM0144 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. Its PCR reaction system is as follows:

[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 isolated st...

Embodiment 3

[0038] Example 3 Tieback test

[0039] 1. Strain cultivation: transfer the tested strains stored at -80°C to YMA liquid culture, and cultivate to logarithmic phase. The growth of the strains is detected by a nucleic acid protein analyzer, and the OD value is used to calculate the bacterial content; among them, YMA ( Yeast MannnitolAgar) liquid medium composition is: take by weighing mannitol 10 g, MgSO 4 ∙7H 2 O 0.2g, NaCl 0.1g, yeast powder 3g, K 2 HPO 4 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 epidermis for 3 minutes, rins...

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Abstract

The invention discloses rhizobium YZM0144 and application thereof. The rhizobium YZM0144 is obtained by collecting rhizobium in soil of Youxi area of Fujian Province through a MO peanut novel variety,the rhizobium is separated and purified from fresh nodules, a 16s rDNA partial sequence of a strain is determined and identified as a novel strain of Bradyrhizobium by a phylogenetic analysis, and the rhizobium is named as Funong No. 4. The rhizobium YZM0144 proves that the rhizobium has the advantages of high-efficiency nodulation and strong nitrogen-fixing ability through laboratory hydroponicreturning experiments and field returning experiments, inoculation of the rhizobium can increase peanut biomass and yield, and the rhizobium can be applied to acid soil in South China as one of the measures for the routine cultivation of peanuts.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a rhizobia YZM0144 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 t...

Claims

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Application Information

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