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Colloidal lysobacter OH17 and application thereof in plant growth

A colloidal lysobacterium, plant growth technology, applied in the direction of plant growth regulators, plant growth regulators, applications, etc., can solve problems such as unspecified metabolic pathways, and achieve the goal of improving stress resistance, reducing massive accumulation, and improving growth Effect

Inactive Publication Date: 2018-09-28
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the invention does not clarify the specific metabolic pathways involved in ACC deaminase in Agrobacterium tumefaciens, the changes in plant rhizosphere ethylene content of ACC deaminase in the process of promoting plant growth, and the strains in the process of plant stress resistance. the role of

Method used

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  • Colloidal lysobacter OH17 and application thereof in plant growth
  • Colloidal lysobacter OH17 and application thereof in plant growth
  • Colloidal lysobacter OH17 and application thereof in plant growth

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Concentrate 10L of Lyobacter colloidis OH17 fermentation supernatant to 300mL by vacuum distillation, mark ACC according to the reaction system in Table 1, place it at 37°C and 180rpm for 2 hours, and use high-performance liquid chromatography to analyze the supernatant after reaction. For detection, the detection instrument used is Shimadzu MS-2020, the chromatographic column is C-18column, the detection wavelength is 350nm, the detection temperature is 35°C, acetonitrile is used as the mobile phase, and the mobile phase detection conditions are shown in Table 2. The reference substance is the ACC standard labeled with Marfey's reagent.

[0036] Table 1 Marfey's reagent labeled ACC reaction system

[0037] Reagent

Volume / μL

Supernatant

100

1M sodium bicarbonate

40

Marfey's reagent

100

[0038] Table 2 Conditions for high performance liquid chromatography work

[0039] time

Acetonitrile volume concentration ...

Embodiment 2

[0043] 30mL OD 600 Lyobacter colloidis OH17 fermentation broth of ≈2 was ultrasonically crushed (Ultrasonic Homogenizer, Scientz-IID) for 20 minutes, centrifuged at 6000rpm for 5 minutes, the supernatant of the cell disruption liquid was absorbed and concentrated to 5 mL by vacuum distillation, and the concentrated cell disruption liquid was taken The supernatant was labeled with 2-ketobutyric acid according to the reaction system in Table 3, and reacted at 80° C. for 1 hour. Use high performance liquid chromatography to detect the supernatant after the reaction. The detection instrument is Shimadzu MS-2020, the chromatographic column is C-18column, the detection wavelength is 450nm, and acetonitrile is used as the mobile phase. The mobile phase detection conditions are shown in Table 4 below.

[0044] Table 3 labeled 2-ketobutyric acid reaction system

[0045] Reagent

[0046] Table 4 Conditions for high performance liquid chromatography work

[0047] tim...

Embodiment 3

[0050] The amino acid sequences of EC 3.5.99.7 (Pseudomonas entomophila) and EC 2.5.1.48 (Cicerarietinum) were respectively compared with BioEdit (Version 7.0.9.0) to find the corresponding functional genes in Lysobacter colloidus OH17 strain.

[0051] Compare the result as Figure 5 with Figure 7 shown, where Figure 5 For Peg_1256 and from Phytophthora sojae (UniProt: G5AFQ7), Azospirillum lipoferum (UniProt: Q1G755), Bradyrhizobiumdiazoefficiens (UniProt: Q89XR6) and Pseudomonas entomophila (UniProt: C3VP48) in the ACC deaminase amino acid sequence homology comparison, 1256peg_ It has 62% amino acid homology. Figure 7 Homology alignment of Peg_5321 with cystathionine γ-synthase amino acid sequences from Cicer arietinum (UniProt: L0EK12), Mycobacterium ulcerans (UniProt: AOPKT3) and Helicobacter pylori (UniProt: Q1M0P5), peg_5321 has 59% amino acids homology.

[0052] Further protein domain analysis of the found genes revealed that Peg_1256 has a protein domain charact...

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Abstract

The invention discloses colloidal lysobacter OH17 and application thereof in plant growth. The colloidal lysobacter OH17 contains cystathionine gamma-synthase genes for synthesis of 1-aminocyclopropane-1-carboxylic acid (ACC) and ACC deaminase genes. The ACC deaminase genes can decompose the ACC into alpha-ketobutyric acid and ammonia, the ACC is the synthesis precondition of ethylene, generationof the ethylene is effectively reduced through ACC deaminase, and thus the adaptive capacity of plants under adversity stress is improved. Compared with the prior art, the generation amount of the ethylene is effectively decreased through the genes for ACC synthesis in the colloidal lysobacter OH17, the survival rate of the plants is increased, the pest-resistant capability is enhanced, and the plant growth condition is improved.

Description

technical field [0001] The invention belongs to the field of microbial genetic engineering, and in particular relates to Lysobacter colloidus OH17 and its application in plant growth. Background technique [0002] With the development of the economy, people's damage to the environment, soil salinization, heavy metal pollution, drought and organic pollution are becoming more and more serious. Environmental stress leads to a large accumulation of stress ethylene in plants, and excessive ethylene can cause plant growth retardation or even death. . How to effectively improve the stress resistance of plants and increase the yield of crops has become a widespread concern. [0003] Lysobacteria widely exist in nature, and are often found in soil and water. The bacteria have no flagella and have a high G+C content in the genome. They belong to the family Xanthomonadaceae and belong to the genus Lysobacter. Hydrolytic enzymes and small molecule antibacterial substances have bacteri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P21/00C12R1/01
CPCA01N63/00C12N1/205C12R2001/01
Inventor 拉博德·佩德罗刘凤权凌军朱润杰贾艺凡
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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