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Pseudomonas and separating method and application thereof

A technology of Pseudomonas and Pseudomonas strains, which is applied in the field of microorganisms, can solve the problem that the streak bacteria do not show resistance, and achieve a significant antagonistic effect

Active Publication Date: 2018-08-24
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of production, no rice varieties that are immune to spot disease have been found, and rice varieties that are generally resistant to bacterial blight of rice do not show resistance to spot disease.

Method used

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  • Pseudomonas and separating method and application thereof
  • Pseudomonas and separating method and application thereof
  • Pseudomonas and separating method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the acquisition of Pseudomonas 1257

[0040] 1. Soil source

[0041] Rhizosphere Soil of Chinese Cabbage in Vegetable Field in Fengxian District, Shanghai

[0042] 2. Screening of bacterial strains

[0043] (1) Soil sample collection

[0044] Zig-shaped 5-point sampling method for soil sample collection: collect 200g soil samples at each point, mix the soil samples at 5 points taken according to the Z-shape evenly, take 200g by quartering method and put them into a sterilization bag as a soil sample . Three soil samples were collected from each plot as replicates. Record the time, place and type of sampling. The collected soil samples were stored in a refrigerator at 4°C for use in bacterial isolation.

[0045] (2) Isolation of bacteria

[0046] Plate dilution method: Weigh 10g of each soil sample into a sterile conical conical flask, add 90mL of sterile water, then place at 200rpm, shake in a shaker at 28°C for 15min, take out the conical flask and...

Embodiment 2

[0052] Embodiment 2, the identification of the 16S rRNA gene of Pseudomonas 1257

[0053] Genomic DNA of strain 1257 was extracted using primer: 27F5'-AGAGTT TGA TCCTGGCTCAG-3' and 1492R5'-TACGGCTACCTTGTTACGACTT-3'; using the extracted DNA as a template, perform PCR amplification to obtain the target fragment. The PCR reaction system is:

[0054]

[0055] The reaction mixture was mixed gently and then briefly centrifuged to perform PCR amplification. The PCR reaction conditions are:

[0056] 94°C for 3min,

[0057] 94°C 30sec, 56°C 30sec, 72°C 90sec, 30 cycles,

[0058] 72℃8min,

[0059] 25°C Forever.

[0060] Take 5 μL of the above-mentioned PCR amplification product, and use 1% agarose gel for electrophoresis detection (see image 3 ), the PCR stock solution was sent to Platinum Biotechnology (Shanghai) Co., Ltd. for sequencing, and its base sequence is shown in SEQ ID No.1.

[0061] The sequencing results were analyzed using DNA Star and compared with BLAST on t...

Embodiment 3、1257

[0063] Physiological and biochemical identification of embodiment 3, 1257

[0064] The physiological and biochemical characteristics of Pseudomonas 1257 of the present invention are: it can produce fluorescent pigment, can secrete arginine dihydrolase and protease, and can assimilate glucose, mannose, mannitol, gluconate, capric acid, apple Acid, citric acid, phenylacetic acid, see Table 1. Can utilize 62 kinds of carbon sources such as Tween 40, L-arabinose, D-fructose, α-D-glucose, etc. It cannot hydrolyze indole, escin, and β-galactosidase, and cannot reduce nitric acid to nitrite, see Table 2.

[0065] Table 1 Physiological and biochemical characteristics of 1257 - enzyme activity, carbon source assimilation

[0066]

[0067] +: Positive reaction; -: Negative reaction;

[0068] Table 2 Physiological and biochemical characteristics of 1257—acid production using carbon source

[0069]

[0070]

[0071]

[0072] +: Positive reaction; -: Negative reaction; W: W...

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Abstract

The invention relates to pseudomonas and a separating method and application thereof. Pseudomonas strains are preserved by China Center for Type Culture Collection in April 10 , 2018, wherein thepreservation name is Pseudomonas entomophila 1257, and the preservation number is CCTCC No:M 2018189. Pseudomonas has an obvious effect of inhibiting bacterial leaf streak, and meanwhile, pseudomonasalso has the effect of inhibiting xanthomonas oryzae pv.oryzae and magnaporthe oryzae.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of pseudomonas and its isolation method and application. Background technique [0002] Xanthomonas oryzae pv.oryzicola (Xoc) is a pathogenic species of Xanthomonas oryzae, which causes bacterial leaf streak (BLS), and is an important bacterial disease on rice. The disease mostly occurs in rice planting areas such as Asia and Africa (Zhang Rongsheng, Liu Yongfeng, Chen Zhiyi. Screening, evaluation and application of antagonistic bacteria to rice bacterial spot disease[J]. Chinese Journal of Biological Control, 2011, 27(04): 510-514.; Rongsheng Zhang, Zhiyi Chen, Yongfeng Liu.Advance in Rice Bacterial Leaf StreakResearches[J].Agricultural Science&Technology,2015,16(02):298–305+310.), especially in tropical and subtropical regions ( Adhikari T.B., and Mew T.W., Pathogenic variability in Xanthomonas compestriis pv. oryzicola, IRRN., 1985, 10(1), 16), such...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A01N63/00A01P1/00C12R1/38
CPCA01N63/00C12N1/02C12N1/20C12N1/205C12R2001/38
Inventor 邹丽芳李生樟杨瑞环美丽·吾尼尔别克杨小菲张翠萍陈晓斌陈功友
Owner SHANGHAI JIAO TONG UNIV
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