Saline-alkaline tolerant bacillus siamensis strain and production method and application of live bacteria preparation of saline-alkaline tolerant bacillus siamensis strain

Abstract

The present invention discloses a saline-alkaline tolerant bacillus siamensis strain and a production method and an application of a live bacteria preparation of the saline-alkaline tolerant bacillussiamensis strain. The strain is isolated from the rhizosphere soil of rice and deposited in China General Microbiological Culture Collection Center on June 22, 2020, and has a preservation number of CGMCC No.20119. The bacillus siamensis PAPM-15 strain has strong saline-alkaline tolerance and strong ability to produce NH<3> and ACC deaminase, and the live bacteria preparation can effectively promote growth of rice, also has a relatively high resistance to rice bakanae disease caused by fusarium fujikuroi, and can be used to produce biological fertilizers, especially the biological fertilizersspecific for rice in saline-alkali soil.

Description

technical field

[0001] The invention relates to a production method and application of a Bacillus siamese strain and a live bacterial preparation thereof, belonging to the field of agricultural biotechnology. Background technique

[0002] Rice is one of the main food crops in my country, with a wide range of planting areas. In addition to being used as a staple food, rice has many other uses. Therefore, the quality and yield of rice are inseparable from national food security. However, judging from the actual situation of rice planting, there are many factors that reduce rice production. Among them, bakanae disease is a common factor that affects rice production. 20%, severe cases can reduce production by more than 50%.

[0003] Rice bakanae disease, also known as excessive growth disease, can occur from the seedling stage to the heading stage, and is mainly transmitted by infected seeds. Fusarium fujikuroi is the main pathogen of rice bakanae disease, Fusarium graminearu...

Examples

Embodiment 1

[0030] Example 1: Screening of Bacillus siamese PAPM-15

[0031] (1) Screening of saline-alkali-tolerant strains

[0032] Bacillus siamese PAPM-15 was isolated from rice rhizosphere soil. The soil was sampled in moderately saline-alkali soil in Bincheng District, Binzhou City, Shandong Province in September 2018. The specific separation method is as follows: mix the soil sample evenly, weigh 5g, put it into a conical flask filled with 95mL sterile water and 10 glass beads, shake it at 37°C and 180rpm for 30min. Take 1 mL of soil suspension for 10 -1 -10 -7 Serial concentration gradient dilution, and then take 10 -5 、10 -6 、10 -7 The three dilutions were spread on the plate containing the selection medium, and cultured upside down at 37°C for 2 days. Single colonies were picked and streaked onto plates containing selection medium, and cultured upside down at 37°C for 2 days. Pick a single colony and transfer it to the slant of the preservation medium test tube, culture ...

Embodiment 2

[0048] The identification of embodiment 2 Bacillus siamese PAPM-15

[0049] (1) Morphological and physiological and biochemical characteristics

[0050] The morphological characteristics of the Bacillus siamese PAPM-15 strain are: cultivated on NA medium at 37°C for 48h, the diameter of the colony is about 0.5-0.8mm, white and opaque, round, with irregular edges, rough surface, bulges, and wrinkles ; Bacteria are rods, such as figure 1 shown. The NA medium is nutrient agar medium, and its formula is: beef extract 3g, peptone 10g, NaCl 5g, agar 20g, water 1000mL, pH 7.0.

[0051] Physiological and biochemical characteristics of the Bacillus siamese PAPM-15 strain: Gram-positive bacilli; positive catalase test, positive starch hydrolysis test, positive nitrate reduction test, positive malonate utilization test, positive gelatin liquefaction test , The methyl red test was negative, the D-glucose fermentation test was positive, the D-mannitol fermentation test was positive, and...

Embodiment 3

[0055] The preparation of embodiment 3 bacillus siamese PAPM-15 microbial agent

[0056] The preparation method of described bacillus siamese PAPM-15 strain microbial agent comprises the following steps:

[0057] 1) Activation of strains: Transfer the low-temperature-preserved Bacillus siamese PAPM-15 strains to the slant of NA medium test tubes, and culture them at 32-37° C. for 24-36 hours for activation. The NA medium is nutrient agar medium, and its formula is: beef extract 3g, peptone 10g, NaCl 5g, agar 20g, water 1000mL, pH7.0.

[0058] 2) Triangular flask seed preparation: Scrape the activated Bacillus siamese PAPM-15 lawn with an inoculation loop, inoculate it in NB medium, and culture it at 32-37° C. for 24-36 hours. The NB medium is a nutrient broth medium, and its formula is: 3 g of beef extract, 10 g of peptone, 5 g of NaCl, 1000 mL of water, and pH 7.0.

[0059] 3) Preparation of seed tank strains: Transfer the seeds of the triangular flask to a 10L seed tank eq...

technical field

[0001] The invention relates to a production method and application of a Bacillus siamese strain and a live bacterial preparation thereof, belonging to the field of agricultural biotechnology. Background technique

[0002] Rice is one of the main food crops in my country, with a wide range of planting areas. In addition to being used as a staple food, rice has many other uses. Therefore, the quality and yield of rice are inseparable from national food security. However, judging from the actual situation of rice planting, there are many factors that reduce rice production. Among them, bakanae disease is a common factor that affects rice production. 20%, severe cases can reduce production by more than 50%.

[0003] Rice bakanae disease, also known as excessive growth disease, can occur from the seedling stage to the heading stage, and is mainly transmitted by infected seeds. Fusarium fujikuroi is the main pathogen of rice bakanae disease, Fusarium graminearu...

Examples

Embodiment 1

[0030] Example 1: Screening of Bacillus siamese PAPM-15

[0031] (1) Screening of saline-alkali-tolerant strains

[0032] Bacillus siamese PAPM-15 was isolated from rice rhizosphere soil. The soil was sampled in moderately saline-alkali soil in Bincheng District, Binzhou City, Shandong Province in September 2018. The specific separation method is as follows: mix the soil sample evenly, weigh 5g, put it into a conical flask filled with 95mL sterile water and 10 glass beads, shake it at 37°C and 180rpm for 30min. Take 1 mL of soil suspension for 10 -1 -10 -7 Serial concentration gradient dilution, and then take 10 -5 、10 -6 、10 -7 The three dilutions were spread on the plate containing the selection medium, and cultured upside down at 37°C for 2 days. Single colonies were picked and streaked onto plates containing selection medium, and cultured upside down at 37°C for 2 days. Pick a single colony and transfer it to the slant of the preservation medium test tube, culture ...

Embodiment 2

[0048] The identification of embodiment 2 Bacillus siamese PAPM-15

[0049] (1) Morphological and physiological and biochemical characteristics

[0050] The morphological characteristics of the Bacillus siamese PAPM-15 strain are: cultivated on NA medium at 37°C for 48h, the diameter of the colony is about 0.5-0.8mm, white and opaque, round, with irregular edges, rough surface, bulges, and wrinkles ; Bacteria are rods, such as figure 1 shown. The NA medium is nutrient agar medium, and its formula is: beef extract 3g, peptone 10g, NaCl 5g, agar 20g, water 1000mL, pH 7.0.

[0051] Physiological and biochemical characteristics of the Bacillus siamese PAPM-15 strain: Gram-positive bacilli; positive catalase test, positive starch hydrolysis test, positive nitrate reduction test, positive malonate utilization test, positive gelatin liquefaction test , The methyl red test was negative, the D-glucose fermentation test was positive, the D-mannitol fermentation test was positive, and...

Embodiment 3

[0055] The preparation of embodiment 3 bacillus siamese PAPM-15 microbial agent

[0056] The preparation method of described bacillus siamese PAPM-15 strain microbial agent comprises the following steps:

[0057] 1) Activation of strains: Transfer the low-temperature-preserved Bacillus siamese PAPM-15 strains to the slant of NA medium test tubes, and culture them at 32-37° C. for 24-36 hours for activation. The NA medium is nutrient agar medium, and its formula is: beef extract 3g, peptone 10g, NaCl 5g, agar 20g, water 1000mL, pH7.0.

[0058] 2) Triangular flask seed preparation: Scrape the activated Bacillus siamese PAPM-15 lawn with an inoculation loop, inoculate it in NB medium, and culture it at 32-37° C. for 24-36 hours. The NB medium is a nutrient broth medium, and its formula is: 3 g of beef extract, 10 g of peptone, 5 g of NaCl, 1000 mL of water, and pH 7.0.

[0059] 3) Preparation of seed tank strains: Transfer the seeds of the triangular flask to a 10L seed tank eq...

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