CN1624108A - Bdellovibrio sp. and its application - Google Patents

Abstract

A Bdellovibrio sp. S-23 (CGMCC No.1045) able to dissolve microcystis is disclosed. It can be used to treat the nutrients-enriched water in river or lake.

Description

A kind of Bdellovibrio and application thereof

Technical field

The present invention relates to a kind of bacterium and application thereof, particularly a kind of Bdellovibrio and application thereof.

Background technology

" Bdello " in the Bdellovibrio (Bdellovibrio) has leech or " bloodsucker's " meaning.What wherein study comparatively in detail is bacteriophagic Bdellovibrio (Bdellovibrio bacteriouorus is called for short the Bd bacterium).Bacteriophagic Bdellovibrio was found from Kidney bean leaf blight pseudomonas by Stolp and Petiold first in 1962, the uniqueness that this bacterium had " parasitism " and " phagocytosis " biological nature are paid close attention to by the microbial world scholar, countries in the world are separated to the Bd bacterium in succession from soil, natural water, seawater later on, and have carried out deep research.The Bd bacterium has the life cycle of a two-phase, they have a motion of not growing to attack nursery stage in phase and the cell that does not move, and are very active in their motion of attack phase, constantly go to seek the host and attack, along with Bdellovibrio is adsorbed onto on the host cell wall, they begin to enter nursery stage.Then Bdellovibrio passes adventitia and peptidoglycan layer and rests between the host cell pericentral siphon and breed, and loses flagellum simultaneously.The Bd bacterium is that a class is attacked, infected, the bacterial parasite of cracking host bacterium, and the effect of similar phage is arranged.Bd bacterium thalline is less, is the Gram-negative bacteria that is about 0.25-0.4 μ m * 0.8 μ m * 1.2 μ m, is arcuation or shaft-like, and the thalline afterbody has a long flagellum, and it is extremely active to move.

States such as Europe, the United States are comparatively deep aspect the fundamental research of Bd bacterium, aspects such as research emphasis is mainly synthetic at biological characteristics, the albumen of Bd bacterium, DNA combination, bacteriolyze material.Recent years, China scientific research personnel was done a large amount of work aspect Bd bacterium control poultry, livestock intestinal tract bacterial infectious disease, had formed a kind of new technology of biological control.

Microcystis aeruginosa (Microcystis) is the microorganism of Cyanophyta, Cyanophyceae, Chroococcales, is the arch-criminal who causes water to pollute, the Microcystin that contains in the Microcystis aeruginosa even suitable with the elapid snake venom toxicity of same purity.Along with the raising of industrialization degree, the eutrophication in lake grows in intensity in recent years, and lake, various places outburst again and again is the wawter bloom of advantage algae kind with the blue-green algae, and wherein the wawter bloom with the Dian Chi is comparatively serious.The Dian Chi wawter bloom is to be the blue-green alga bloom of absolute predominance algae kind with the Microcystis aeruginosa, and country and Yunnan Province have dropped into lot of manpower and material resources and administered the Dian Chi, but still fails controlled and solution.Administer at present the water that Microcystis aeruginosa causes and pollute the general technologies such as little flocculation, ozone, gac that adopt, but since the problem of cost and the efficient of algicide own remove the algae effect and be still waiting further raising.Microorganism control algae technology is a very promising improvement wawter bloom technology,

The innovation and creation content

The purpose of this invention is to provide a kind of Bdellovibrio that can dissolve Microcystis aeruginosa.

The Bdellovibrio that can dissolve Microcystis aeruginosa provided by the present invention is Bdellovibrio (Bdellovibrio sp.) S-23, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on November 25th, 2003, preserving number is CGMCC № 1045.

The water sample separation of Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 in Beijing University Wei Xiu Yuan pond obtains.

Experiment showed, that Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 can dissolve Microcystis aeruginosa effectively.The biology control algae factor as Microcystis aeruginosa, both can be applied to separately and carry out molten algae in the contaminated water, also can cooperate other algae-lysin factor (as other algae-lysin factors such as PdY3 of dissolving anabena), synergy, effectively administering body eutrophication, particularly the blue-green alga bloom aspect plays a significant role, and market potential is huge.

The present invention will be further described below in conjunction with accompanying drawing.

Description of drawings

Fig. 1 is the negative staining electromicroscopic photograph of Bdellovibrio S-23

Fig. 2 infects the electromicroscopic photograph of the starting stage of Microcystis aeruginosa for Bdellovibrio S-23

Fig. 3 is the electromicroscopic photograph that infects the Bdellovibrio S-23 that breeds in the Microcystis aeruginosa cell

Fig. 4 is for Bdellovibrio S-23 and contrast the result who respectively polluted water is handled

Embodiment

Separation, bacteria characteristic, nutritional mode, the molten algae mode of embodiment 1, Bdellovibrio (Bdellovibrio sp.) S-23

1, the separation of Bdellovibrio (Bdellovibrio sp.) S-23

Water sampling in Beijing University Wei Xiu Yuan pond, water sample is added in the Microcystis aeruginosa of pure culture, illumination cultivation, after 3-5 days, the flavescence of algae liquid, with molten algae drop on the aseptic filter paper sheet, cover in the double-deck algae plate that Microcystis aeruginosa is paved into, molten algae plaque can occur around the filter paper, As time goes on plaque constantly enlarges, and gets the agar block of plaque edge, render in the pure culture Microcystis aeruginosa, after treating molten algae,, obtain Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 with the molten algae liquid of 0.45um membrane filtration.

2, the bacteria characteristic of Bdellovibrio (Bdellovibrio sp.) S-23

(1) morphological specificity of Bdellovibrio (Bdellovibrio sp.) S-23

The size of Bdellovibrio is between 0.22um and 0.45um, and it can be by the filter membrane of 0.45um, and can not be by the filter membrane of 0.22um, its morphological structure as shown in Figure 1, thalline is arcuation.Carry out flagella staining and gramstaining according to a conventional method, the result shows that this thalline afterbody has a long flagellum, belongs to Gram-negative bacteria.

(2) physiological and biochemical property of Bdellovibrio (Bdellovibrio sp.) S-23

S-23 is a kind of bacterium of parasitics, can not on common substratum, independently grow, and in the experiment, be with the host of Microcystis aeruginosa as it always, it obtains nutrition by parasitizing in the Microcystis aeruginosa cell.When temperature was lower than 24 ℃, S-23 lost the function that infects Microcystis aeruginosa.

(3) nutritional mode of Bdellovibrio (Bdellovibrio sp.) S-23

The nutritional mode of Bdellovibrio S-23 is divided into two vegetative phases: i.e. infective moving period and noninfective nursery stage.Moving period, Bdellovibrio S-23 was in the Microcystis aeruginosa extracellular, had flagellum, can specificity ground infect Microcystis aeruginosa, the effect by a series of lytic enzymes enters in the Microcystis aeruginosa cell, loses flagellum simultaneously, breeds to absorb frustule nutrition, carries out the transition to nursery stage; Regenerate flagellum after breeding the certain period, cracking Microcystis aeruginosa cell discharges Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045, goes to infect new Microcystis aeruginosa again.

(4) the molten algae mode of Bdellovibrio (Bdellovibrio sp.) S-23

By getting different molten algae Bdellovibrio (Bdellovibrio sp.) the S-23 CGMCC № 1045 molten algae liquid in period, under oily mirror, observe, when finding just to have begun molten algae, the small-particle that some brown are arranged in the Microcystis aeruginosa, the bacterium of arcuation is being assembled on many Microcystis aeruginosas next door, prolongation along with the time, has only the small-particle that brown is arranged in the Microcystis aeruginosa less and less, and almost do not have the brown particle in the molten algae liquid more than molten three week of algae, bracket fungus in the solution is more and more simultaneously, the experiment proved that the brown particle is the vegetation of Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045.Can judge that according to the bacteria characteristic of Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 it is a kind of new Bdellovibrio.

Embodiment 2, polluted water is handled with Bdellovibrio S-23 of the present invention

Get the Microcystis aeruginosa of 3 bottles of 60ml pure cultures respectively, frustule concentration is 10 ⁸Individual/ml, one bottle in contrast, and (concentration of S-23 is 10 respectively to add the molten algae liquid of Bdellovibrio S-23 Microcystis aeruginosa of 1ml in other two bottles of Microcystis aeruginosas ⁹Individual/as ml), to have added the algae liquid flavescence of S-23 after 3 days, frustule all disintegrates after the week, solution becomes clear (result as shown in Figure 4).

Claims (3)

1, Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 and mutant and varient.

2, the application of Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 in administering body eutrophication.

3, the application of Bdellovibrio (Bdellovibrio sp.) S-23 CGMCC № 1045 in administering blue-green alga bloom.

Images