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High-efficiency rhizobium meliloti strain with stress resistance and growth promoting performances and application of rhizobium meliloti

A technology of Rhizobium alfalfa and Sinorhizobium, applied in the direction of microorganism-based methods, chemicals for biological control, bacteria, etc., can solve the problem of low nitrogen fixation efficiency, low affinity between Rhizobium and alfalfa varieties, and competitive nodulation Insufficient capacity and other problems, to achieve the effect of improving alfalfa quality, high-efficiency field competition nodulation ability, strong dissolution of inorganic phosphorus and organic phosphorus

Active Publication Date: 2017-07-28
XINJIANG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problems of poor affinity between inoculated rhizobia and alfalfa varieties, poor competitive nodulation ability and low nitrogen fixation efficiency, and to provide a strain with strong stress resistance, An excellent rhizobia strain with high growth-promoting and nodulation-competitive ability and high nitrogen fixation efficiency

Method used

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  • High-efficiency rhizobium meliloti strain with stress resistance and growth promoting performances and application of rhizobium meliloti

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Effect test

Embodiment 1

[0026] Isolation, purification and identification of embodiment 1 bacterial strain

[0027] 1. Isolation and purification of strains

[0028] Fresh root nodules were collected from 12 alfalfa artificial grasslands in southern and northern Xinjiang (counties, group farms). Select a robust plant with a large, plump red nodule on the lateral root, rinse the nodule with sterile water, soak it in 75% ethanol for 5 min, and then wash it with 0.1% mercuric chloride (HgCl 2 ) after disinfection for 5 min, rinse with sterile water, take the last sterile water cleaning solution and apply it to a YMA plate and culture it for 72 h for sterility test. Use sterile tweezers to crush a single root nodule, use an inoculation loop to pick up the bacterial suspension and streak it on the YMA plate medium, culture it at a constant temperature of 28 ℃ for 2~4 d, purify the strain several times, pick a single colony and transfer it to the slant, Stored at a low temperature of 4 ℃, and named the o...

Embodiment 2

[0042] Example 2 The back graft test and matching test of Sinorhizobium alfalfa XGL026

[0043] The backgraft test of Sinorhizobium meliloti XGL026 adopts the agar tube method: select large, plump and undamaged alfalfa seeds of No. % mercuric chloride solution for surface disinfection for 5 min, and finally washed with sterile water 4-6 times, 5 min each time. Put the sterilized seeds on the agar plate, germinate at 28 ℃ for 2 d, place the germinated seeds with consistent growth on the inclined surface of the 1% agar nitrogen-free culture medium test tube, and inoculate when the first true leaf appears (each tube of inoculum solution 1 mL), repeat 4 tubes, set up a blank control without inoculation, culture in the light incubator, 10~30 d after emergence, observe and record the nodulation results. Tieback test results (see Figure 4 , Figure 5 ) showed that the strain XGL026 showed good nodulation ability, the average number of nodules reached 5.5  / plant, and the nodulated...

Embodiment 3

[0044] Example 3 Determination of stress resistance of Sinorhizobium alfalfa XGL026

[0045] The stress resistance of Sinorhizobium meliloti XGL026 was mainly tested for salt tolerance, acid and alkali tolerance, and NH tolerance. 4 + Ability, drought tolerance and growth temperature range determination. The YMA medium was used as the basal medium, and the YMA liquid medium or solid medium plate cultured at 28 ℃ with pH 7.0 was used as the positive control, and each treatment was repeated 3 times. Salt resistance, acid and alkali resistance, NH resistance 4 + , and the determination of drought tolerance were to activate the strains on YMA medium plates, inoculate them in YMA liquid medium, and culture them at 28 ℃, 160 rpm / min for 2 d (OD 600 Value 0.800) as the inoculum, inoculate 2% of the inoculum into the culture solution of each treatment, 28 ℃, 160 rpm / min shaking culture for 4 d, UV-2000 spectrophotometer at 600 nm wavelength to measure the bacteria of each treatmen...

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Abstract

The invention discloses a high-efficiency rhizobium meliloti strain with stress resistance and growth promoting performances and application of rhizobium meliloti. The strain is sinorhizobium meliloti XGL026, is separated and purified from fresh alfalfa rhizobium meliloti and is collected in China General Microbiological Culture Collection Center on January 21, 2013, the collection address of the strain is Institute of Microbiology, Chinese Academy of Sciences, 3#, Yard 1#, Beichen West Road, Chaoyang District, Beijing, and the collection number is CGMCC NO. 7179. The high-efficiency rhizobium meliloti strain XGL026 disclosed by the invention is high in stress resistance, has the capabilities of dissolving inorganic phosphorus and organic phosphorus and secreting IAA and is an excellent rhizobium meliloti strain with high matching affinity with main cultivated alfalfa varieties in Xinjiang, high competitive nodulation ability and high nitrogen-fixing efficiency. When applied to an alfalfa production area in Xinjiang, the strain disclosed by the invention has high applicability, high nodulation ability and effects of improving the alfalfa forage yield and improving the quality, the overall growth performance of alfalfa plants is enhanced, and the contrast difference from non-inoculated strains reaches an obvious level.

Description

technical field [0001] The invention relates to a biological strain resource, in particular to a root nodule strain which can symbiotically fix nitrogen with alfalfa and has excellent stress resistance, growth promotion performance and outstanding nodulation ability in the field. Background technique [0002] Rhizobia are a class of Gram-negative rod-shaped bacteria that can infect legumes under suitable conditions and perform symbiotic nodulation and nitrogen fixation with them, converting free nitrogen in the air into combined nitrogen that plants can use. Rhizobium-legume symbiosis is the most important biological nitrogen fixation system in nature, and the amount of nitrogen fixation accounts for about 65%-70% of the total biological nitrogen fixation. Alfalfa ( Medicago sativa L.) is the forage crop with the largest planting area in the world, and is known as the "king of grass". It has a strong root system, strong adaptability, high grass yield, and good nutrition...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01P21/00C12R1/01
CPCC12N1/205C12R2001/01
Inventor 罗明马霞韩剑何媛郭海玉皇甫彩娟李卫军
Owner XINJIANG AGRI UNIV
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