Tea tree growth-promoting rhizobacterium Serratia marcescens and application thereof

Abstract

The invention discloses a tea tree growth-promoting rhizobacterium of which the classification designation is Serratia marcescens. The strain number is JW-CZ2. The Serratia marcescens is collected by China Center for Type Culture Collection, the collection number is CCTCC M 2015095, and the collection date is March 10th, 2015. The invention also discloses application of the Serratia marcescens in promoting tea tree growth. The JW-CZ2 strain has high solubility for insoluble phosphates, such as tricalcium phosphate, magnesium phosphate and iron phosphate. The JW-CZ2 strain has IAA (indoleacetic acid) secretion capacity and ACC deaminase producing capacity. When a tea tree seedling plant is inoculated in the JW-CZ2 microbial inoculant, the microbial inoculant can obviously promote the growth of the tea tree seedling. Therefore, the tea tree growth-promoting rhizobacterium Serratia marcescens can provide an excellent strain resource for future development of special microbial fertilizers for tea trees.

Description

technical field

[0001] The invention belongs to the technical field of microbial fertilizers in the field of biological fertilizers, and in particular relates to Serratia marcescens, a growth-promoting bacterium in the rhizosphere of tea trees and an application thereof. Background technique

[0002] Tea tree [Camellia sinensis (L.) O.Kuntze] belongs to the Camellia family Camellia genus Camellia. It is a perennial, woody and evergreen plant. One of the crops (Lin Sheng et al., 2012). According to statistics, as of 2012, my country's tea garden area was 35.29 million mu, accounting for about 1.15% of the country's cultivated land area, ranking first in the world (Department of Planting Management, Ministry of Agriculture, 2012). The tea leaves made from the nutritious young shoots of tea trees are recognized as one of the three major beverages (tea, coffee, cocoa) in the world because of their rich nutrients and good health care effects. However, picking new green and nutr...

Examples

Embodiment 1

[0024] Example 1: Determination of phosphorus-dissolving ability of JW-CZ2 strain.

[0025] Phosphate Solubilizing Medium A: Glucose 10g, Ca 3 (PO 4 ) 2 5g, MgCl 2 5g, KCl 0.2g, MgSO 4 ·7H 2 O 0.25g, (NH 4 ) 2 SO 4 0.1g, distilled water 1000mL, pH 7.0.

[0026] Phosphate-dissolving medium B: using iron phosphate (FePO 4 ), magnesium phosphate (Mg 3 (PO 4 ) 2 ) instead of tricalcium phosphate (Ca 3 (PO 4 ) 2), the other components and contents are the same. Inoculate the twice activated JW-CZ2 strain into NB medium (3.0g of beef extract, 10.0g of peptone, 5.0g of sodium chloride, 1000mL of distilled water, 18.0g of agar, pH7.2-7.4), and culture at 30°C for 18-24 Make JW-CZ2 seed solution afterward, inoculate in the 100mL Erlenmeyer flask that contains 50mL NBRIP medium by 1% inoculum amount, take the NBRIP medium that inoculates NB medium as control (CK), each process 3 repetitions, After shaking culture at 30°C and 200r / min for 72h, the fermentation broth was ...

Embodiment 2

[0028] Example 2: Qualitative analysis of IAA produced by JW-CZ2 strain.

[0029] After inoculating the JW-CZ2 strain in NA liquid medium and culturing for 12 hours (28°C, 120r / min), adjust the OD600 value to 0.05 with sterile water, and take 0.1mL of the test bacteria suspension to receive the In the King culture solution containing 100mg / L tryptophan, each triangular flask was filled with 50mL culture solution, repeated 3 times, and the culture solution with 0.1mL sterile water was used as the blank control, and placed together at 28 Cultivate for 12 days in a temperature-controlled shaker with a rotational speed of 120 r / min. Take 50 μL of the test bacteria suspension and 50 μL of the blank control respectively, add 50 μL of colorimetric solution, put it at room temperature, and observe the color change within 15 minutes. If all three repetitions turn red, it means that IAA can be secreted; the darker the color, the better the secretion of IAA. The number is large; all 3 r...

Embodiment 3

[0030] Example 3: Quantitative analysis of IAA produced by JW-CZ2 strain.

[0031] The tested strains were inoculated in king medium shake flasks and cultured for 3 days. First, measure the OD of the bacterial suspension by spectrophotometry 600 value. Then the bacterial suspension was centrifuged at 10000r / min for 10min, and 1mL of the supernatant was added to an equal volume of Salkowski colorimetric solution, and kept in the dark for 30min for color development. The OD was determined by spectrophotometry 530 value. Calculation of bacterial concentration OD 600 When the value is 1, the amount of IAA secreted by bacteria in a unit volume of bacterial suspension. The standard curve was prepared by gradient dilution of analytically pure IAA.

[0032] Preparation of colorimetric solution: Preparation of S1 colorimetric solution: weigh FeCl 3 12g was dissolved in 300mL distilled water, then slowly added 429.7mL 98% H 2 SO 4 , after cooling down to 1L, the measuring range...

technical field

[0001] The invention belongs to the technical field of microbial fertilizers in the field of biological fertilizers, and in particular relates to Serratia marcescens, a growth-promoting bacterium in the rhizosphere of tea trees and an application thereof. Background technique

[0002] Tea tree [Camellia sinensis (L.) O.Kuntze] belongs to the Camellia family Camellia genus Camellia. It is a perennial, woody and evergreen plant. One of the crops (Lin Sheng et al., 2012). According to statistics, as of 2012, my country's tea garden area was 35.29 million mu, accounting for about 1.15% of the country's cultivated land area, ranking first in the world (Department of Planting Management, Ministry of Agriculture, 2012). The tea leaves made from the nutritious young shoots of tea trees are recognized as one of the three major beverages (tea, coffee, cocoa) in the world because of their rich nutrients and good health care effects. However, picking new green and nutr...

Examples

Embodiment 1

[0024] Example 1: Determination of phosphorus-dissolving ability of JW-CZ2 strain.

[0025] Phosphate Solubilizing Medium A: Glucose 10g, Ca 3 (PO 4 ) 2 5g, MgCl 2 5g, KCl 0.2g, MgSO 4 ·7H 2 O 0.25g, (NH 4 ) 2 SO 4 0.1g, distilled water 1000mL, pH 7.0.

[0026] Phosphate-dissolving medium B: using iron phosphate (FePO 4 ), magnesium phosphate (Mg 3 (PO 4 ) 2 ) instead of tricalcium phosphate (Ca 3 (PO 4 ) 2), the other components and contents are the same. Inoculate the twice activated JW-CZ2 strain into NB medium (3.0g of beef extract, 10.0g of peptone, 5.0g of sodium chloride, 1000mL of distilled water, 18.0g of agar, pH7.2-7.4), and culture at 30°C for 18-24 Make JW-CZ2 seed solution afterward, inoculate in the 100mL Erlenmeyer flask that contains 50mL NBRIP medium by 1% inoculum amount, take the NBRIP medium that inoculates NB medium as control (CK), each process 3 repetitions, After shaking culture at 30°C and 200r / min for 72h, the fermentation broth was ...

Embodiment 2

[0028] Example 2: Qualitative analysis of IAA produced by JW-CZ2 strain.

[0029] After inoculating the JW-CZ2 strain in NA liquid medium and culturing for 12 hours (28°C, 120r / min), adjust the OD600 value to 0.05 with sterile water, and take 0.1mL of the test bacteria suspension to receive the In the King culture solution containing 100mg / L tryptophan, each triangular flask was filled with 50mL culture solution, repeated 3 times, and the culture solution with 0.1mL sterile water was used as the blank control, and placed together at 28 Cultivate for 12 days in a temperature-controlled shaker with a rotational speed of 120 r / min. Take 50 μL of the test bacteria suspension and 50 μL of the blank control respectively, add 50 μL of colorimetric solution, put it at room temperature, and observe the color change within 15 minutes. If all three repetitions turn red, it means that IAA can be secreted; the darker the color, the better the secretion of IAA. The number is large; all 3 r...

Embodiment 3

[0030] Example 3: Quantitative analysis of IAA produced by JW-CZ2 strain.

[0031] The tested strains were inoculated in king medium shake flasks and cultured for 3 days. First, measure the OD of the bacterial suspension by spectrophotometry 600 value. Then the bacterial suspension was centrifuged at 10000r / min for 10min, and 1mL of the supernatant was added to an equal volume of Salkowski colorimetric solution, and kept in the dark for 30min for color development. The OD was determined by spectrophotometry 530 value. Calculation of bacterial concentration OD 600 When the value is 1, the amount of IAA secreted by bacteria in a unit volume of bacterial suspension. The standard curve was prepared by gradient dilution of analytically pure IAA.

[0032] Preparation of colorimetric solution: Preparation of S1 colorimetric solution: weigh FeCl 3 12g was dissolved in 300mL distilled water, then slowly added 429.7mL 98% H 2 SO 4 , after cooling down to 1L, the measuring range...

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