CN113549573A

CN113549573A - Burkholderia strain and application thereof

Info

Publication number: CN113549573A
Application number: CN202110775626.XA
Authority: CN
Inventors: 刘彩霞; 徐婷; 王瑞; 陈隆升; 何之龙; 唐炜; 陈永忠
Original assignee: Hunan Academy of Forestry
Current assignee: Hunan Academy of Forestry
Priority date: 2021-07-08
Filing date: 2021-07-08
Publication date: 2021-10-26
Anticipated expiration: 2041-07-08
Also published as: CN113549573B

Abstract

The invention discloses a burkholderia strain and application thereof, wherein the strain is classified and named as: burkholderia bacterium (B.), (C.)Burkholderiasp.) YCYP9 with the following accession number: CCTCC NO: M2021733. The strain has the function of dissolving inorganic phosphorus and organic phosphorus simultaneously, and can be used for preparing phosphorus-dissolving preparations. Meanwhile, the strain also has the function of promoting the growth of the camellia oleifera seedlings, and can be used for preparing the camellia oleifera seedling growth promoter.

Description

Burkholderia strain and application thereof

Technical Field

The invention relates to a Burkholderia strain and application thereof, in particular to a Burkholderia strain YCYP9 with a phosphorus dissolving effect and application thereof in growth of oil tea seedlings.

Background

Camellia oleifera (Camellia olefera) is a general name of oil species with high oil content and production value in seeds of Camellia (Camellia) of Theaceae, is a special high-quality woody oil tree species in China, and is called four major woody oil tree species in the world together with oil palm, coconut and olive oil. The existing camellia oleifera cultivation area of 6800 ten thousand acres in China is mainly distributed in 14 provinces (regions) in the south, the total industrial value reaches 1160 hundred million yuan, the camellia oil squeezed from camellia oleifera seeds has unique flavor and rich nutrition, the fatty acid composition is reasonable, the content of unsaturated fatty acid reaches 90%, the content of oleic acid is more than 80%, erucic acid and behenic acid which are difficult to absorb by human bodies are not contained, cholesterol which can cause vascular sclerosis due to the increase of the blood pressure of human bodies is not contained, and the oil quality is superior to other edible oil such as olive oil, and the camellia oleifera is ideal edible oil. The camellia oleifera has wide adaptability and barren resistance, is one of good economic forest species in low mountains, hills and hills in the south, does not compete with grains and cotton for land, has the characteristics of one-year planting and multi-year benefit, has the high yield period of 80 years, and plays an important role in rural economic and ecological construction in the south of China.

The large seedling breeding is one of the main means for improving the survival rate of the camellia oleifera planting, the vegetative growth of the large camellia oleifera seedlings is completed in a seedling raising field, a basic tree structure exists during field planting, the fruiting period entering time can be shortened, and the waste of land resources is reduced^[4]. Meanwhile, in the conventional nursery stock outplanting and excavating process, root systems of the conventional nursery stocks are damaged, new roots need to be revived and germinate again, the serious damaged survival rate is low, the root systems of the large seedlings grow in the container all the time, the root system groups of the nursery stocks cannot be damaged during field planting, the large seedlings can continue to grow after being planted, the seedling reviving period is not provided, the survival rate is almost one hundred percent, and therefore the advantages of the large seedlings in the camellia planting process are obvious. In the process of large seedling breeding, the microbial inoculum is widely used due to green and high efficiency, but the microbial inoculum for large seedling breeding of the camellia oleifera has been reported at present, and no applicable finished product is obtained.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a burkholderia strain and application thereof.

In order to achieve the purpose, the technical scheme provided by the invention is as follows:

the invention is used in lakeCollecting oil tea rhizosphere soil in multiple areas of southern province, and using PKO culture medium (glucose 10.0g, (NH)₄)₂SO₄0.5g，MgSO₄·7H₂O 0.3g，NaCl 0.3g，KCl 0.3g，FeSO₄·7H₂O 0.03g，MnSO₄·H₂O 0.03g，Ca₃(PO4)₃ 2.0g，CaCO₃1.0g of agar and 20.0g of agar) and screening and dissolving inorganic phosphorus strains; monkina medium (glucose 10.0g, (NH) was used₄)₂SO₄ 0.5g，MgSO₄·7H₂O 0.3g，NaCl 0.3g，KCl 0.3g，FeSO₄·7H₂O 0.03g，MnSO₄·H₂0.03g of O, 0.2g of lecithin, CaCO₃1.0g, yeast powder 0.5g and agar 20.0 g). In the research, 1 strain with the function of dissolving inorganic phosphorus and organic phosphorus is found, and is named as: burkholderia sp (Burkholderia sp.) YCYP9, which was deposited at the China center for type culture Collection (Wuhan, China) at 6/18 th 2021, with the strain accession number: CCTCC NO: M2021733. The strain has the function of dissolving inorganic phosphorus and organic phosphorus simultaneously, and can be used for preparing phosphorus-dissolving preparations. Meanwhile, the strain also has the function of promoting the growth of the camellia oleifera seedlings, and can be used for preparing the camellia oleifera seedling growth promoter.

The invention is further illustrated below:

in the invention, a qualitative experiment of phosphorus dissolving capacity is measured by a phosphorus dissolving ring method, strains are respectively point-connected in a culture medium, each strain is repeated for three times, and after 3 days of culture, the phosphorus dissolving ring D and the diameter D of a bacterial colony are measured. When the ratio of D/D is more than 1, the inoculated strain has phosphorus dissolving capacity. YCYP9 strain was spotted on a phosphorus-containing plate, and after 3D of culture, the diameter (D) of the strain was 1.30. + -. 0.12cm, the diameter (D) of the phosphorus-solubilizing ring was 3.70. + -. 0.37cm, and the content (D/D) of the dissolved phosphorus was 2.85. + -. 0.38. And (3) measuring the dissolved phosphorus strength by adopting an ammonium molybdate colorimetric method, and inoculating the strain with the dissolved phosphorus ring into a liquid culture medium overnight to activate the strain. Preparing liquid culture medium containing calcium phosphate and yolk lecithin, subpackaging into 50mL, adjusting pH to 7.0, inoculating 2%, inoculating at 28 deg.C and 160r/min for 7 d. Centrifuging the bacterial liquid at 10000r/min for 15-18min, and taking the supernatant to measure the content of available phosphorus in the culture solution by using a flow analyzer. Sign boardPreparing a quasi solution: phosphorus standard stock solution (1000 mg/L): drying at 105 deg.C for 2 hr₂PO₄4.394g were metered to 1L. Phosphorus standard working solution (100 mg/L): 10mL of the stock solution was dissolved in a 100mL volumetric flask and shaken well for use. Standard curve: respectively taking 0mL, 1 mL, 2 mL, 4 mL, 6 mL, 8 mL, 10mL and 20mL of phosphorus-labeled working solution, adding into a 100mL volumetric flask, and fixing the volume. The concentrations are 0, 1, 2, 4, 6, 8, 10 and 20mg/L respectively. The research finds that YCYP9 is used for inorganic phosphorus Ca₃(PO₄)₂The dissolved amount of (A) is 31.82 mg/L; the dissolving amount of the organic phosphorus (egg yolk lecithin) is 4.25 mg/L. The identification result of the strain is Burkholderia sp, the strain is applied to camellia oleifera seedlings, and an LB culture medium (tryptone 10g/L, yeast extract 5g/L, sodium chloride 10g/L and pH adjusted to about 7.4) is used for culturing the strain, so that the viable count of bacterial liquid reaches 10⁷cfu/mL, 50mL of bacterial liquid was added to 2-year-old container seedlings, and 15 seedlings were treated each with LB medium as a control. After three months of growth, the height of the contrast treated seedlings is 62.50 +/-4.32, the ground diameter is 7.98 +/-1.45, and the number of spring shoots is 14.13 +/-1.14; the height of the camellia oleifera seedlings treated by the YCYP9 is 75.78 +/-5.54, the ground diameter is 8.81 +/-1.99, the number of spring shoots is 24.11 +/-2.22, the height of the camellia oleifera seedlings is increased by 21.25% by the YCYP9, the ground diameter is increased by 10.40%, and the number of spring shoots is increased by 70.69%.

Drawings

FIG. 1 is the effect of the strain on the growth amount of Camellia oleifera seedlings.

Detailed Description

The selective medium and the colony morphology difference are utilized to primarily screen 14 strains (YCWP1-14) dissolving inorganic phosphorus strains and 16 strains (YCYP1-16) dissolving organic phosphorus strains. And (3) inoculating the phosphorus-dissolving strains into a phosphorus-containing flat plate, culturing for 3d, and observing the sizes of colonies and phosphorus-dissolving rings, wherein experimental results show that only 7 strains have phosphorus-dissolving rings, namely the phosphorus-dissolving rings have the phosphorus-dissolving capacity, and the phosphorus-dissolving capacities of YCYP9 and YCYP15 are stronger (Table 1).

TABLE 1 qualitative study of phosphorus solubilizing ability (D/D)

The phosphorus dissolving capacity of 7 strains is determinedThe measurement shows that the Ca-containing inorganic phosphorus has dissolution₃(PO₄)₂The strains with the highest dissolving capacity are YCYP9 and the dissolving amount of phosphorus is 31.82 mg.L^-1(ii) a The number of the strains capable of dissolving organic phosphorus (egg yolk lecithin) was only 5, and the strain having the highest dissolving power was YCYP9, and the amount of the dissolved organic phosphorus was 4.25 mg.L^-1(see Table 2).

TABLE 2 quantitative study of phosphorus solubilizing ability

The selected strains are subjected to seedling growth promotion effect study by taking camellia oleifera abel 'Xianglin 210' cutting seedlings as study objects, the study results are shown in figure 1, YCYP9 remarkably promotes the growth of camellia oleifera abel seedling height and spring shoots compared with a control, the mean value of the camellia oleifera abel seedlings after being treated by YCYP9 is 75.78, the ground diameter mean value is 8.81, the number of spring shoots is 24.11, YCYP9 enables the camellia oleifera abel seedling height to be increased by 21.25% compared with the control, the ground diameter to be increased by 10.40%, and the number of spring shoots to be increased by 70.69%.

Claims

1. A burkholderia strain is characterized in that the taxonomic name of the strain is as follows: burkholderia bacterium (B.), (C.)Burkholderiasp.) YCYP9 with the following accession number: CCTCC NO: M2021733.

2. Use of the strain of claim 1 for the preparation of a phosphorus solubilizing preparation.

3. The use of claim 2, wherein the phosphorus-dissolving formulation is a formulation that dissolves both inorganic phosphorus and organic phosphorus.

4. The use of the strain of claim 1 in the preparation of a camellia oleifera seedling growth promoter.