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Klebsiella oxytoca and application thereof to promotion of growth of codonopsis

A technology of Klebsiella and Codonopsis, applied in plant growth regulators, microorganism-based methods, chemicals for biological control, etc., to improve drought resistance, reduce MDA content, and increase soluble protein content Effect

Active Publication Date: 2018-09-28
长治学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Feng Weiwei and others screened out ACC deaminase bacteria with various growth-promoting properties (nitrogen fixation, phosphorus dissolution, and IAA production) from the rhizosphere of the halophyte medicinal plant Limonium sinense in coastal tidal flats of Jiangsu, which has certain research value (Feng Weiwei et al. , 2016); Tian Lei et al. obtained a strain JJ8-3 with high ACC deaminase activity from Panax ginseng C.A.Mey endophytes, and has phosphorus-solubilizing properties, nitrogen-fixing potential and siderophilic production ability, and proved that it can significantly promote the growth of ginseng seeds and roots (Tian Lei et al., 2014); but so far, there are few studies on the growth-promoting bacteria of Ludangshen, and the inoculation of ACC deaminase-producing bacteria Ludangshen Research on membrane lipid peroxidation, soluble protein content, and protective enzyme activity under drought stress is rarely reported, and clarifying these physiological indicators is important for inoculating ACC deaminase-producing bacteria under drought stress to improve the drought resistance of Ludangshen significance

Method used

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  • Klebsiella oxytoca and application thereof to promotion of growth of codonopsis
  • Klebsiella oxytoca and application thereof to promotion of growth of codonopsis
  • Klebsiella oxytoca and application thereof to promotion of growth of codonopsis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Determination of ACC deaminase activity of strain LDS17.

[0026] (1) Insert the strain LDS17 into 50mL TSB medium, shake and culture at 28°C 200r / min for 24h, pipette 1mL bacterial suspension into 50mL TSB medium, expand culture at 28°C 200r / min for 24h, centrifuge at 4°C 9000r / min 10min, and collect the bacteria.

[0027] (2) Use DF liquid medium without adding [(NH 4 ) 2 SO4 ] Centrifuge and wash the bacteria twice, resuspend in 25mL ADF medium, culture at 28°C 200r / min for 48h to induce the production of ACC deaminase, centrifuge the bacterial suspension at 4°C 9000r / min for 10min, discard the supernatant, The bacterial cells were collected and the weight of the bacterial cells was recorded, and then the bacterial cells were centrifuged and washed twice with 0.1 mol / L Tris-HCl buffer solution (pH=7.6).

[0028] (3) The bacteria were resuspended in 600 μL of 0.1 mol / L Tris-HCl buffer (pH=8.5) containing 30 μL of toluene, and shaken rapidly for 30 s to b...

Embodiment 2

[0036] Example 2: Growth-promoting properties of the LDS17 strain

[0037] (1) Determination of siderophilic ability

[0038] Spot-inoculate the LDS17 strain on the CAS medium, and culture it at 28°C for 48-72 hours. If there is a yellow-green halo around the strain, it proves that the strain has the ability to produce siderophiles.

[0039] (2) Determination of the ability to dissolve inorganic phosphorus

[0040] Spot-inoculate the LDS17 strain on NBRI-BPB medium and culture at 28°C for 48-72 hours. If there is a transparent circle around the strain, it proves that the strain has the ability to dissolve inorganic phosphorus. Pick the bacterial strain with an inoculation loop, inoculate it in a 100mL Erlenmeyer flask containing 20mL NBRI-BPB medium, and use the uninoculated blank NBRI-BPB medium as a control, 30°C, 180r / min shaking culture for 3d, the fermentation broth ( Centrifuge at 4°C, 12000r / min) for 10min, measure the OD of the supernatant at a wavelength of 600nm 6...

Embodiment 3

[0058] Embodiment 3: LDS17 greenhouse pot experiment:

[0059] After activating the strain LDS17, use an inoculation loop to pick a small amount of bacteria and inoculate it into a 100mL Erlenmeyer flask containing 50mL NA medium (3g beef extract, 10g peptone, 5g sodium chloride, 1000mL distilled water, pH 7.2-7.4), 28 Cultivate with shaking at 200r / min for 24h. The fermentation broth (4°C, 10000r / min) was centrifuged for 10min to collect the bacteria, rinsed with sterile normal saline for 3 times, adjusted the bacterial suspension with sterile normal saline (10 8 cfu / mL) to make the inoculant. Inoculate the seedlings of Codonopsis pilosula, take the same amount of sterile saline as the control, and the inoculation volume is 5mL / plant. 10 replicates per treatment were placed in the greenhouse for unified management, the light was 12h / day, and watering was timely.

[0060] The growth of Ludangshen 30d and 60ds after inoculation is shown in Table 3. It can be seen that inocul...

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Abstract

The invention discloses klebsiella oxytoca. The classification name is klebsiella oxytoca, the strain number is LDS17, the klebsiella oxytoca is collected in the China Center for Type Culture Collection, the strain collection number is CCTCC NO: M 2017583, and the collection date is October 16, 2017. The strain can grow normally in a culture medium with 1-aminocyclopropane-1-carboxylic acid (ACC)as a sole nitrogen source through enrichment culture, and has relatively strong ACC deaminase activity and other growth promoting properties (IAA and siderophore production, HCN production, NH3 production, nitrogen fixation and phosphorus solubilization); after a potted young seedling of the codonopsis is inoculated with a microbial agent produced by the strain, and results show that the microbialagent can significantly promote the growth and the development thereof; inoculation of the strain LDS17 under drought stress effectively promotes the growth and the drought resistance of the codonopsis. Therefore, an excellent strain resource can be provided for developing a special microbial fertilizer for the codonopsis in the future.

Description

technical field [0001] The invention belongs to the technical field of microbial fertilizers in the field of biological fertilizers, and in particular relates to a Klebsiella oxytoca producing ACC deaminase in the rhizosphere of Ludangshen and its application. Background technique [0002] Codonopsis pilosula, a traditional Chinese medicine, is the dried root of Codonopsis pilosula, which has the functions of enhancing immunity, dilating blood vessels, lowering blood pressure, improving microcirculation, and enhancing hematopoietic function (Guan Linjing et al., 2015). Codonopsis pilosula is one of the commonly used bulk Chinese medicinal materials in my country, and the authentic medicinal material is Shanxi Lu Codonopsis pilosula. Ludangshen is mainly produced in Jincheng, Changzhi and other areas in the southeast of Shanxi Province, especially in the eastern mountainous area of ​​Lingchuan County, Jincheng City, which accounts for 70% of the total area of ​​the county. Th...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/14A01P21/00C12R1/22
CPCC12N9/14C12Y305/99007C12N1/205C12R2001/22
Inventor 任嘉红韩明跃张桂萍吴伟晨晓霓晋婷婷常欣
Owner 长治学院
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