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A kind of rhizobia famb126 and its application

A FAMB126, rhizobia technology, applied in the field of microorganisms, to achieve the effect of improving total nitrogen content, high nodulation rate and strong nitrogen fixation ability

Inactive Publication Date: 2020-07-31
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 famb126 and its application
  • A kind of rhizobia famb126 and its application
  • A kind of rhizobia famb126 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation and purification of Rhizobium FAMB126

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

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

[0025] (3) Cut into two halves on a flame-sterilized glass slide, clamp half of the tumor with sterile tweezers, scribe the incision facing the surface of the YMA (Table 1) medium, invert it and incubate at 28°C.

[0026] (4) After the bacteria grow, use a sterile toothpick to scrape a small amount of rhizobia colonies from the plate, dilute with sterile water, and streak the culture on the plate again. After 3 days, observe the colony situation, until 15 Day...

Embodiment 2

[0033] Example 2 Rhizobium 16s rDNA sequence sequencing

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

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

[0036]

[0037] Obtain 12 reference strain sequences from the NCBI (GenBank) database, and use the software BioEdit and MEGA6 to analyze the isolated strains and reference strains. 16s rDNA partial sequence analysis to construct a phylogenetic tree of the isolated strain and the reference strain (attached figure 2 ). Therefore, ...

Embodiment 3

[0038] Example 3 Tie back test

[0039] 1. Strain culture:

[0040] 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 with a nucleic acid protein analyzer, and the bacterial content was calculated using the OD value. The formula of YMA (Yeast Mannnitol Agar) liquid medium is: Weigh 10 g of mannitol, MgSO 4 ∙7H 2 O 0.2g, NaCl 0.1 g, yeast powder 3 g, K 2 HPO 4 0.25g, KH 2 PO 4 0.25 g, CaCO 3 3 g (added when storing), 15 g of agar dissolved in 1 L of pure water.

[0041] 2. Sterilization treatment:

[0042] Wrap the 2.5L plastic buckets, beakers, tweezers, petri dishes, filter paper, and glass rods required for the test, and put the vermiculite in a fresh-keeping bag, sterilize at 121°C for 30 min. YMA liquid culture medium and distilled water are bottled and sterilized at 121°C for 20 min.

[0043] Put the seeds of peanuts into 1g / L HgCl 2 Sterilize the epidermis in the middle for ...

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Abstract

The invention discloses a rhizobia FAMB126 and its application. The strain is obtained from the rhizobia in the soil of Anxi area, Fujian Province by Minhua No. 8 peanut variety, and the rhizobia is separated and purified from fresh nodules, and the strain of 16s The rDNA partial sequence was determined, and through phylogenetic analysis, it was determined to be Bradyrhizobium ( Bradyrhizobium ) of the new strain and named it Funong 2. The rhizobia FAMB126 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 FAMB126 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 specifically relates to a rhizobia FAMB126 and its applications. Background technique [0002] Rhizobium ( rhizobium ) Is a type of gram-negative bacteria widely distributed in the soil. It can infect the roots or stems of legumes to form root nodule or stem nodule, and fix the N in the air as a symbiont 2 Become NH that can be absorbed by plants 4 + . The symbiotic system between legumes and rhizobia occupies an extremely important position in the entire biological nitrogen fixation. It is estimated that about 100 million tons of nitrogen are fixed from the air on the earth's surface every year, and about 55 million tons of nitrogen are fixed by legumes and rhizobia. Rhizobia and legumes form a symbiotic nitrogen fixation system. In addition to meeting the plant’s own nitrogen needs, it can also provide excess nitrogen to non-legumes mixed or intercropped with them, such as non-legumes mixed with ...

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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