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Biological antagonistic bacterial strain for controlling postharvest disease of fruit and vegetable, and preparation method and application thereof

A postharvest disease and antagonistic bacteria technology, applied to the biological antagonistic bacteria strains for preventing postharvest diseases of fruits and vegetables, its preparation and application fields, can solve the problems of polluting the environment, not easy to degrade, and odor of fungicides, etc., to achieve the promotion of plant growth, The effect of low cost and excellent disease resistance

Inactive Publication Date: 2014-07-30
FARM PROD STORAGE & FRESHENING INST SHANXI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many fungicides have peculiar smell, pollute the environment, and are not easy to degrade. Therefore, it is increasingly urgent to find substitutes or partial substitutes for fungicides with low toxicity, high efficiency, environmental friendliness and low residue

Method used

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  • Biological antagonistic bacterial strain for controlling postharvest disease of fruit and vegetable, and preparation method and application thereof
  • Biological antagonistic bacterial strain for controlling postharvest disease of fruit and vegetable, and preparation method and application thereof
  • Biological antagonistic bacterial strain for controlling postharvest disease of fruit and vegetable, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: Preparation of biologically antagonistic bacteria strain Enterobacter cowii B-6-1

[0034] There are a large number of microorganisms that can inhibit pathogens in tomato fruit surface. The strains are now isolated from the tomato surface and wounds. The specific method is as follows: take 3 tomatoes, put them in a container containing 0.2M phosphate buffer, wash them in a 100rpm rotary oscillator for 10 minutes, and discard the washing solution. Then use the same method for a second wash (before the second wash, ultrasonically treat the tomato for 30 seconds), and keep the lotion. Then serially dilute the secondary washing solution (10, 10 2 、10 3 and 10 4 times), spread the diluted solution on LB solid medium, 0.1mL per plate, culture at 28°C for 24-48h until a single colony grows, randomly select a single colony according to different colony characteristics, and put it on the LB solid medium After repeated streaking, pick the purified single colony ...

Embodiment 2

[0035] Embodiment 2: Identification of biologically antagonistic bacterial strain Enterobacter cowanii B-6-1

[0036] The 16S rDNA sequence of the strain is shown in SEQ ID NO: 1, and the phylogenetic tree constructed based on 16S rDNA is shown in the appendix figure 1 .

[0037] The rpoB gene sequence of the strain is shown in SEQ ID NO: 2, and the phylogenetic tree constructed based on the rpoB gene is shown in the appendix figure 2 .

[0038] The identification method of the strain is as follows:

[0039] 1. PCR primers

[0040]16SrDNA forward primer 27F: 5'-AGA GTT TGA TCC TGG CTC AG-3' (see SEQ ID NO: 3),

[0041] 16SrDNA reverse primer 1541R: 5'-AAG GAG GTG ATC CAC CC-3' (see SEQ ID NO: 4),

[0042] RopB gene forward primer CM7-f: 5'-AACCAGTTCCGCGTTGGCCTG-3' (see SEQ ID NO: 5),

[0043] RopB gene reverse primer CM7-r: 5'-CCTGAACAACACGCTCGGA-3' (see SEQ ID NO: 6);

[0044] 2. Cell culture

[0045] After re-culturing the freeze-dried strains, they were separated b...

Embodiment 3

[0071] Embodiment 3: to the biocontrol effect of tomato gray mold

[0072] The preparation concentrations were 0 (control), 1×10 7 cfu / mL, 1×10 8 cfu / mL, 1×10 9 cfu / mL, 1×10 10 cfu / mL of Enterobacter cowii bacteria liquid, after the surface of tomato fruit was sterilized by 2% NaClO, two 5mm×5mm wounds were made on each fruit thorn, and the above four concentrations of Enterobacter cowii B-6-1 suspension were inoculated on the wounds respectively 50μL, after 2h, inoculate 1X10 6 Spores / mL Botrytis cinerea suspension was 30 μL, and the above-mentioned treated fruits were kept moist at 25°C, and the incidence was counted after 7 days. The experiment was repeated three times with 10 fruits per treatment.

[0073] The morbidity statistics are attached image 3 As shown, with the increase of concentration, the incidence of tomato gray mold decreased. The incidence of CK botrytis was 100%; at 1×10 7 When cfu / mL, the incidence rate is 75.33%, 1×10 8 cfu / mL is 57.11%, 1×10 9...

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Abstract

Belonging to the field of microorganisms and biological control, the invention relates to a biological antagonistic bacterium, and a preparation method and application thereof. The bacterial strain is named as Enterobacter cowanii B-6-1, and belongs to Enterobacter Enterobacteriaceae. The main morphological characteristics include rod shape, short rod-like shape, length of 0.8-1.2 mum, width of 1-1.6 mum, Gram-negative bacterium, white colonies, smooth surface, and good growth at room temperature. The bacterial strain provided by the invention has excellent disease resistance performance, and the best formula comprising antagonistic bacterium liquid with concentration more than 1*10<8> cfu / mL and additional 50 m mol / L Na2SiO3 achieves the best disease resistance effect; therefore, the invention has great value to the development of fruit and vegetable farming.

Description

technical field [0001] The invention relates to a biological antagonist in the field of microorganisms and biological control, its preparation method and its application, in particular to the application of the biological antagonist in the prevention and control of post-harvest diseases of fruits and vegetables. Background technique [0002] The loss caused by fungal rot of fruits and vegetables after harvest is very huge. At present, chemical fungicides are one of the main means to control postharvest fungal diseases of fruits and vegetables. However, the long-term use of chemical fungicides has increased their residual levels and led to the emergence of resistant pathogens. For example, many fungicides can control the occurrence of Penicillium and green mold of citrus, but in recent years, due to the extensive use of fungicides, the acquired resistance of Penicillium digitatum and Penicillium italicum has become one of the main methods of postharvest disease control. Big...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23B7/155C12R1/01
Inventor 施俊凤孙常青王潇冉王春生白云凤郭志利
Owner FARM PROD STORAGE & FRESHENING INST SHANXI ACADEMY OF AGRI SCI
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