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 fixatio

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

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Experimental program
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Example Embodiment

[0026] Example 1 Isolation, purification and identification of strains

[0027] 1. Isolation and purification of strains

[0028] Fresh nodules were collected from 12 (county, group farms) alfalfa artificial grasslands in southern and northern Xinjiang. Choose a large, full red nodule on the main and lateral roots of a vigorously growing plant, wash the nodules with sterile water, soak in 75% ethanol for 5 min, and then use 0.1% mercury (HgCl) 2 ) After disinfection for 5 min, rinse with sterile water, take the last sterile water cleaning solution, spread on YMA plate and culture for 72 h for sterility test. Use sterile tweezers to break a single nodule, use an inoculating loop to pick the bacterial suspension and streak it on the YMA plate medium, culture for 2~4 d at a constant temperature of 28 ℃, purify the strain multiple times, pick a single colony and transfer it to the slope. Store at 4 ℃ at low temperature, and name one strain obtained as XGL026.

[0029] The composition o...

Example Embodiment

[0042] Example 2 Back-joining test and matching test of Sinorhizobium alfalfa XGL026

[0043] The back-joining test of Sinorhizobium alfalfa XGL026 adopts the agar tube method: select large, plump and undamaged Xinmu No. 1 alfalfa seeds, soak them in 95% ethanol for 5 min, pour the ethanol, and add the mass volume fraction to 0.1 Disinfect the surface with% mercury solution for 5 min, and finally wash 4~6  times with sterile water for 5 min each time. The sterilized seeds are placed on the agar plate, and the germination is accelerated at 28 ℃ for 2 d. The germination seeds with consistent growth are placed on the slope of the test tube of 1% agar and nitrogen-free culture solution. When the first true leaf appears, inoculate (each tube of inoculation liquid 1 mL), repeat 4 tubes, set up a blank non-inoculation control, culture in a light incubator, 10~30 d after emergence, observe and record the results of nodulation. Tie back test results (see Figure 4 , Figure 5 ) Showed th...

Example Embodiment

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

[0045] The stress resistance of Sinorhizobium alfalfa XGL026 is mainly salt tolerance, acid and alkali tolerance, and NH tolerance. 4 + Determination of ability, drought tolerance and growth temperature range. The YMA medium was used as the basic medium, and the YMA liquid medium or solid medium plate cultured at pH 7.0 at 28 ℃ was used as a positive control. Each treatment was repeated 3 times. Salt resistance, acid and alkali resistance, NH resistance 4 + The determination of drought tolerance is to activate the strains on YMA medium plates, inoculate them in YMA liquid medium, culture 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 each treatment bacteria The optical density value of the suspension...

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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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