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Rhizobium YZLH133 and application thereof

A technology of rhizobia and bacterial strains, applied in the field of microorganisms, to achieve the effects of increasing total nitrogen content, high nodulation rate, and increasing dry weight

Inactive Publication Date: 2019-03-08
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 YZLH133 and application thereof
  • Rhizobium YZLH133 and application thereof
  • Rhizobium YZLH133 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Isolation and purification of rhizobia YZLH133

[0022] (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.

[0023] (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.

[0024] (3) Cut in half on a flame-sterilized glass slide, hold the half tumor with sterile tweezers, mark the incision on the surface of the YMA (Table 1) medium, incubate at 28°C after inversion.

[0025] (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 again on the plate. Observe the colonies after 3 days until 15 day...

Embodiment 2

[0032] Example 2 of Rhizobium YZLH133 16sr DNA sequence sequencing

[0033] 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:

[0034] Table 2 16s rDNA 2×starMix enzyme reaction system

[0035]

[0036] 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

[0037] Example 3 Tieback test

[0038] 1. Strain culture:

[0039] The test strains stored at -80°C were transferred to YMA liquid culture and cultivated to the logarithmic phase. The growth of the strains was detected by a nucleic acid protein analyzer, and the OD value was used to calculate the bacterial content; YMA (Yeast Mannnitol Agar) liquid culture The base components are: weigh 10 g of mannitol, 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

[0041]Wrap the 2.5L plastic bucket, beaker, tweezers, petri dish, filter paper, and glass rod needed for the test, pack the vermiculite in a fresh-keeping bag, and sterilize at 121°C for 30 minutes. YMA liquid medium and distilled water were packed in bottles, and sterilized at 121°C for 20 minutes.

[0042] Put peanut seeds in 1g / L HgCl 2 Sterilize the epidermis for 3...

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Abstract

The invention discloses a rhizobium YZLH133 and application thereof. The strain is a new strain which is captured from the soil in Youxi District, Fujian Province through Longhua peanut variety, is separated and purified from fresh nodules, is determined to be Bradyrhizobium through phylogenetic analysis by testing part sequences of 16s rDNA of the strain, and is named Funong 3#. A laboratory hydroponic tieback test and a large field tieback experiment prove that the rhizobium YZLH133 disclosed by the invention has the characteristics of efficient nodulation and strong nitrogen fixing capacity; the biomass and the yield of peanuts can be improved by inoculating the rhizobium; and the rhizobium is applicable to acid soil in South China to serve as one of the measures for conventional cultivation of the peanuts.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a rhizobia YZLH133 and its application. 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 that are...

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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