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Sesbania cannabina endophytic bacillus ZH60, complex microbial inoculant and corn seed culture method

A technology of compound bacterial agent and cultivation method, which is applied in the field of agricultural resources and environment, can solve the problems of pollution, soil water pollution, and little research on seed soaking agents, and achieve the effects of improving soil, increasing corn yield, and shortening the growth period of crops

Pending Publication Date: 2022-01-25
SHANDONG AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Most of the soaking agents in previous studies were compounded with different chemical components, and the soaking agents entered the groundwater from the soil, causing soil water pollution and environmental pollution; there were few studies on the preparation of seed soaking agents mixed with natural materials and microorganisms

Method used

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  • Sesbania cannabina endophytic bacillus ZH60, complex microbial inoculant and corn seed culture method
  • Sesbania cannabina endophytic bacillus ZH60, complex microbial inoculant and corn seed culture method
  • Sesbania cannabina endophytic bacillus ZH60, complex microbial inoculant and corn seed culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1, Isolation and identification of endophyte Bacillus sp. ZH60

[0054] (1) Separation and purification method of endophyte Bacillus spp. ZH60: Weigh 0.5 g of S. chinensis seeds, soak them in a water bath at 60°C and stir for 3 min, treat with 75% alcohol for 30 s, rinse with sterile water, and treat with 5% NaClO for 5 min , rinsed 5 times with sterile water. The seeds were soaked in phosphate buffered saline (PBS) for 48h. The homogenate was serially diluted after grinding. Each dilution gradient was spread evenly on LB and Gaw's No. 1 solid plates. Incubate at 30°C. After the colonies were formed, the colonies with different shapes and sizes were picked and transferred to the plate, purified by the plate streak method, and transferred three times.

[0055] (2) Identification of endophyte Bacillus ZH60:

[0056] ①The colony morphology of endophyte Bacillus spp. ZH60 is as follows figure 1 As shown, the colony of this strain is Gram-positive bacteria, rod...

Embodiment 2

[0059] Example 2, Test for IAA and Inorganic Phosphorus Reconciliation by Endophyte Bacillus chinensis ZH60

[0060] (1) Qualitative detection of IAA: Endophytes were cultured in LB liquid medium, transferred to LB liquid medium containing tryptophan (200 mg / L) after 24 hours (1% inoculum volume), at 30°C, 180 r / min Incubate for 48h in a shaking incubator with shaking frequency. Centrifuge, take the supernatant and Salkowski colorimetric solution (0.5mol / L FeCl 3 Solution 10ml, 35% HClO 4 500ml) were mixed in an equal amount on a white porcelain plate, placed in a dark place for 30min, and the color change of the reaction solution was observed, and each sample was set to 3 repetitions. The blank was the mixed solution of the IAA solution and the colorimetric reagent. Certified to contain IAA.

[0061] (2) Quantitative detection of IAA: configure 100 mg / L IAA solution, and then configure it into a series of concentration gradient standard solutions of 0-25.0 mg / L. Take 2ml...

Embodiment 3

[0065] Embodiment 3, the compound bacterial liquid of sapura gum and sapella endophyte Bacillus ZH60

[0066] Prepare 100ml of sapura gum solution with a mass fraction of 1% (w / v) in advance;

[0067] Take the purified Bacillus chinensis ZH60 after purification in Example 1 and culture it in 100ml LB liquid medium at 37°C and 180rpm with shaking overnight to OD 600 = 0.8, the inoculum size is 1%; centrifuge at 4000 rpm for 5 min, discard the supernatant, and resuspend in 1% concentration (w / v) sapura gum solution to the solution OD 600 = 0.8.

[0068] The mass ratio of the sapura gum solution to the endophyte Bacillus sassicae ZH60 can be 1:1, or 1:2, or 3:10 or 1:5.

[0069] The sapella gum powder is soluble in the concentration range below 2%. When the concentration is higher than 2%, flocculent insoluble matter appears at room temperature. When the concentration is lower, the viscosity is low and it is not easy to adhere to the surface of the seeds. Glue solution was tes...

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Abstract

The invention discloses a sesbania cannabina endophytic bacillus ZH60, a complex microbial inoculant and a corn seed culture method. The complex microbial inoculant comprises the sesbania cannabina endophytic bacillus ZH60 and sesbania gum; and the sesbania cannabina endophytic bacillus ZH60 and the complex microbial inoculant are environmentally friendly and non-toxic, and can be used for promoting the germination of corn seeds and improving salt tolerance. According to the sesbania cannabina endophytic bacillus ZH60, the complex microbial inoculant and the corn seed culture method, sesbania germ plasm resources are fully utilized, endosperm sesbania gum extraction and endophyte separation are performed on the sesbania germ plasm resources, the salt tolerance and adhesion of the sesbania gum and the growth promoting property of endogenous bacillus velezensis are combined, and the sesbania cannabina endophytic bacillus ZH60 is colonized in heterologous plant corn, so that the growth promoting and salt resisting effects are achieved. The crop growth period is shortened, and the corn yield is improved.

Description

technical field [0001] The invention relates to the technical field of agricultural resources and the environment, in particular to the endophyte Bacillus ZH60, a compound bacterial agent and a method for culturing corn seeds. Background technique [0002] Soil salinization is an important environmental factor restricting the production of food crops and the efficient use of land resources. In general, salinity stress causes nutrient imbalance, oxidative damage, and reduced yield by increasing the toxicity of sodium ions and osmotic stress. [0003] As a major grain-producing country, the output of corn has ranked first in my country's grain output year after year. Corn is not only used as food for human diet, but also an important feed crop, and also an important raw material for medicine, food and chemical products. Corn is a salt-sensitive crop, and corn yields are lower when grown on saline soils. Seed germination and seedling growth are key stages in the life cycle o...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N25/10A01N63/22A01P21/00C09K17/32A01G22/20A01G31/00C12R1/07C09K101/00
CPCC12N1/20A01N25/10A01N63/22C09K17/32A01G22/20A01G31/00C09K2101/00
Inventor 解志红庞诗琪王丹丹殷志秋王洪凤陈大印
Owner SHANDONG AGRICULTURAL UNIVERSITY
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