CN108587959B - Phyllostachys pubescens endogenous phosphorus-dissolving potassium-dissolving nitrogen-fixing enterobacter aerogenes and application thereof - Google Patents
Phyllostachys pubescens endogenous phosphorus-dissolving potassium-dissolving nitrogen-fixing enterobacter aerogenes and application thereof Download PDFInfo
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Abstract
The invention provides a phyllostachys pubescens endogenous phosphorus-dissolving potassium-dissolving nitrogen-fixing gas productionEnterobacter and application thereof, wherein the bacterium is enterobacter aerogenes (enterobacter aerogenes)Enterobacter aerogenes) CT-B09-2, which has been preserved in China general microbiological culture Collection center in 2017, 7, 3 months, and the preservation numbers are: CGMCC No. 14377. The enterobacter aerogenes in the phyllostachys pubescens has the functions of dissolving phosphorus, potassium and nitrogen, can be colonized in plants, improves the activity of biological enzyme, and has important function on regulating the growth and development of the plants.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a phyllostachys pubescens endogenous phosphorus and potassium dissolving nitrogen fixing enterobacter aerogenes and application thereof.
Background
Endophytic bacteria refer to those bacteria which live in various tissues and organs of healthy plants at a certain stage or all stages of their life history and establish a harmonious association with the plants. The endophytic bacteria of the plant can be colonized and conducted in the plant body for a long time due to the ecological advantages of the endophytic bacteria, is not easily influenced by environmental conditions, has wide biological effects on the growth and development of the plant, disease resistance and adverse environment, is a rare natural biological resource, and has good research and development potential in agricultural production. Various endophytic bacteria are separated from various plants such as bamboos, hot peppers, oranges, poplar trees, tobaccos and dendrobium officinale, corresponding research is carried out on the bacteria which are endogenic to plant tissues and have growth promoting functions such as phosphate solubilizing, potassium solubilizing or nitrogen fixing, and the shengg and the like are separated from the plant bodies to the endophytic bacteria which can degrade organic pollutants or have heavy metal resistance and can promote the growth of the plants.
Mao bamboo (A)Phyllostachys edulis) Belongs to a scattered bamboo species, has the advantages of fast growth, early lumber production, wide application, great economic benefit and the like, and is an important forest resource in south China. According to the statistics of forest resource cleaning at the 6 th time in China, the area of the bamboo forest in China is about 337.20 ten thousand hectares and occupies 47 percent of the area of the bamboo forest in the world. However, the research on the moso bamboos mostly focuses on the aspects of moso bamboo yield theory, soil physicochemical properties, species diversity and the like, in recent years, people such as Liyu and Qizei have respectively researched rhizosphere microorganisms and soil bacteria of the moso bamboos and the Brazikia chinensis, and the selection of a separation culture medium of endophytic bacteria at the roots of the moso bamboos is also explored in summer and winter. However, at present, different tissues of the moso bamboo can be cultured to separate endophytic bacteria, and the endophytic bacteria with the functions of phosphate and potassium dissolution and nitrogen fixation are screenedAnd functional endophytic bacteria with photosynthesis promoting effect have not been reported.
Disclosure of Invention
The invention aims to provide a phyllostachys pubescens endogenous phosphorus and potassium dissolving nitrogen fixing enteroaerobacter aerogenes and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
an enterobacter aerogenes strain with the classification name of enterobacter aerogenes (enterobacter aerogenes) by endogenous phosphate-dissolving potassium-dissolving nitrogen-fixing of moso bambooEnterobacter aerogenes) CT-B09-2, which has been preserved in China general microbiological culture Collection center in 2017, 7, 3 months, and the preservation numbers are: CGMCC No.14377, address No. 3 of Xilu No.1 of Beijing, Chaoyang.
The bacterial colony characteristics and the thallus morphology of the enterobacter aerogenes with the functions of internally decomposing phosphorus, decomposing potassium, fixing nitrogen and producing gas are as follows:
the colony formed after the CT-B09-2 is cultured on an NA flat plate for 24 hours is white, glossy, round, smooth and semitransparent in surface, neat in edge, free of fluidity, negative in gram stain and free of spores; the colony diameter is 4-5 mm.
The physiological and biochemical characteristics of the endogenous phosphorus-dissolving potassium-dissolving nitrogen-fixing enterobacter aerogenes are as follows:
CT-B09-2 catalase reaction is positive, V.P determination is positive, methyl red determination is negative, glucose acidogenesis test is positive, glucose gas evolution test is positive, citrate test is positive, nitrate reduction reaction is positive, starch hydrolysis is positive, oxygen demand determination is negative, indole test is negative, malonic acid determination is positive, H production is negative2Positive in S test.
The 16S rDNA sequence of the endogenous phosphorus-dissolving potassium-dissolving nitrogen-fixing aerogenic enterobacter is compared with the sequence in the GenBank database, and the result shows that CT-B09-2 and the sequence in the GenBank databaseEnterobacter aerogenesOn the same branch, the 16S rDNA sequence andEnterobacter aerogenesthe similarity of (JQ 660204) reaches 99%. Combined with colony morphology, physiological and biochemical characteristics and 16S rDNA sequence analysis, is identified as enterobacter aerogenes: (Enterobacter aerogenes）。
The invention has the advantages that:
the enterobacter aerogenes in the phyllostachys pubescens has the functions of dissolving phosphorus, potassium and nitrogen, can be colonized in plants, improves the activity of biological enzyme, and has important function on regulating the growth and development of the plants.
The microbial inoculum prepared from the phyllostachys pubescens endogenous phosphorus-dissolving potassium-dissolving nitrogen-fixing enteroaerogen of the invention is inoculated to the phyllostachys pubescens seedlings by root irrigation, which can obviously improve the chlorophyll content of the phyllostachys pubescens leaves, improve the photosynthetic rate (Pn), the transpiration rate (Tr) and the stomatal conductance (Gs), and reduce the intercellular CO of the leaves2Concentration (Gs), compared to control, was significantly different. The bamboo is inoculated to the moso bamboo with the degree II through bamboo cavity injection, the activity of superoxide dismutase (SOD), Peroxidase (POD), the content of soluble protein and the content of soluble sugar can be improved, and compared with a control, the difference reaches a remarkable level. Therefore, the invention provides good strain resources for developing the microbial agent bacterial fertilizer special for the moso bamboos in the future.
Drawings
FIG. 1 shows the change in SPAD values of Phyllostachys pubescens leaves after treatment with endophytes.
FIG. 2 change in CAT activity of Phyllostachys pubescens leaves after endophyte treatment.
FIG. 3 change in MDA content of Phyllostachys pubescens leaves after endophyte treatment.
FIG. 4 change in POD activity of Phyllostachys pubescens leaves after endophyte treatment.
FIG. 5 change of SOD activity in Phyllostachys pubescens leaves after treatment with endophytic bacteria.
FIG. 6 change in soluble protein content of Phyllostachys pubescens leaves after treatment with endophytic bacteria.
FIG. 7 change in soluble sugar content of moso bamboo leaves after treatment with endophytic bacteria.
Detailed Description
The present invention is further illustrated by the following examples.
An enterobacter aerogenes strain with the classification name of enterobacter aerogenes (enterobacter aerogenes) by endogenous phosphate-dissolving potassium-dissolving nitrogen-fixing of moso bambooEnterobacter aerogenes) CT-B09-2, which has been preserved in China general microbiological culture Collection center in 2017, 7, 3 months, and the preservation numbers are: CGMCC No. 14377.
Example 1 screening of Enterobacter aerogenes having phosphorus-solubilizing, potassium-solubilizing, nitrogen-fixing effects in Phyllostachys Pubescens
(1) And (3) screening:
82 isolates are separated from I-degree moso bamboos in central production areas of moso bamboos in Fujian province, Wuyi mountain city, Changting county and Lexian county by a gradient dilution method. Purifying the separated endophytic bacteria by a three-region scribing method, judging whether the bacterial strain is purified by microscopic examination, numbering the purified bacteria, picking a single bacterial colony, and transferring the single bacterial colony to an NA inclined plane for storage and standby. Through the determination of the effect of flat-plate phosphate-dissolving potassium-dissolving nitrogen-fixing, 20 endophytic strains with better phosphate-dissolving potassium-dissolving nitrogen-fixing effect are preliminarily screened out, and finally, one endophytic bacterium with good phosphate-dissolving potassium-dissolving nitrogen-fixing effect is screened out and marked as CT-B09-2.
(2) Colony characteristics and colony morphology:
the colony formed after the CT-B09-2 is cultured on an NA flat plate for 24 hours is white, glossy, round, smooth and semitransparent in surface, neat in edge, free of fluidity, negative in gram stain and free of spores; the colony diameter is 4-5 mm.
(3) Physiological and biochemical characteristics:
CT-B09-2 catalase reaction is positive, V.P determination is positive, methyl red determination is negative, glucose acidogenesis test is positive, glucose gas evolution test is positive, citrate test is positive, nitrate reduction reaction is positive, starch hydrolysis is positive, oxygen demand determination is negative, indole test is negative, malonic acid determination is positive, H production is negative2Positive in S test.
(4) And (3) determining the phosphorus-dissolving, potassium-dissolving and nitrogen-fixing capacities:
and respectively inoculating the separated endophytic bacterial strains to an organic phosphorus culture medium, an inorganic phosphorus culture medium, a potassium bacteria culture medium and an Ashby nitrogen-free culture medium plate which are prepared in advance, wherein 4 inoculation points are arranged on each dish, and repeating for 3 times. Culturing at 28 deg.C for 5 days. And respectively observing and recording the growth condition of the strain and the size of a decomposition ring, and determining the phosphate-solubilizing potassium-solubilizing nitrogen-fixing activity of the endophytic bacteria according to the size of the decomposition ring, the diameter of the transparent ring and the diameter of the bacterial colony (D/D value). The larger the decomposition ring is, the larger the D/D value is, the stronger the phosphorus-dissolving, potassium-dissolving and nitrogen-fixing activity is.
Wherein, the organic phosphorus culture medium: glucose 10g, (NH)4)2SO40.5g，NaCl 0.3g，KCl 0.3g，MnSO40.03g，FeSO40.03g, lecithin 0.2g, CaCO35.0g, 0.4 g of yeast extract, 20g of agar, 1000mL of distilled water and pH of 7.0-7.2 (the agar is not added in a liquid culture medium);
inorganic phosphorus culture medium: glucose 10g, (NH)4)2SO40.5g，NaCl 0.3g，KCl 0.3g，MnSO40.03g，FeSO40.03g，MgSO40.3g，CaCO35.0g，Ca3(PO4)25.0g, 0.4 g of yeast extract, 20g of agar, 1000mL of distilled water and pH of 7.0-7.2 (the agar is not added in a liquid culture medium);
potassium bacteria culture medium: sucrose 10.0g, yeast extract 0.5 g, (NH)4)2SO4,1.0g，Na2HPO42.0g，MgSO4·7H2O 0.5g，CaCO31.0 g, 1g of potassium feldspar powder, 15g of agar, 1000mL of distilled water and pH7.0-7.2 (the agar is not added in a liquid culture medium);
ashby nitrogen-free medium: sucrose 10.0g, yeast extract 0.5 g, (NH)4)2SO4,1.0g，Na2HPO42.0g，MgSO4·7H2O 0.5g，CaCO31.0 g, potassium feldspar powder 1g, agar 15g, distilled water 1000mL, pH7.0-7.2 (liquid medium does not add agar).
By measurement, the organophosphorus degrading activity of the enterobacter aerogenes CT-B09-2 is +++, the diameter (D/D) of the transparent circle/colony is 5.05 +/-0.41, the inorganic phosphorus degrading activity is +++, the diameter (D/D) of the transparent circle/colony is 2.12 +/-0.08, the inorganic phosphorus degrading activity is ++, and the diameter (D/D) of the transparent circle/colony is 3.30 +/-0.36. The nitrogen fixation activity is "+ + +".
TABLE 1 preliminary screening of high-efficiency P-K-solubilizing N-fixing strains
D, the diameter of the transparent ring, D, the diameter of the bacterial colony,
"-": no activity (decomposition ring < 10 mm); "+": has activity (decomposition ring: 10-15 mm);
"++": strong activity (decomposition ring: 16-20 mm); "+++": very active (decomposition ring: > 20 mm);
inoculating the bacterial strain with phosphate and potassium dissolving activity in the primary screen into NB culture medium until the concentration of bacterial suspension reaches 108And when cfu/mL is required, respectively inoculating 5mL of the bacterial suspension into 100 mL of an organic phosphorus liquid culture medium, an inorganic phosphorus liquid culture medium and a potassium bacteria liquid culture medium, repeating each treatment for 3 times, and simultaneously taking no inoculation as a control. Centrifuging (4 ℃, 10000 r/min, 15 min) after culturing at 28 ℃ and 160 r/min for 7d, taking supernate, and measuring the effective phosphorus increment (the value after deducting the control) by adopting a molybdenum-antimony colorimetric method, wherein the soluble phosphorus content calculation formula is as follows: p ═ K × V/V1. Wherein, P is the available phosphorus content; k is the phosphorus content (mg/L) of the color development liquid found by a standard curve; v is the volume (mL) of the solution for constant volume during color development; v1 is the volume (mL) of supernatant aspirated at the time of color development. The effective potassium increment (value after subtraction of control) was determined by flame photometry.
By determination, the release of soluble phosphorus from 0.2g/L lecithin by the enterobacter aerogenes CT-B09-2 is 81.77mg/L, which is 53.10 times of that of the control treatment, and the difference is obvious (P is less than 0.05); the soluble phosphorus released from 5.0g/L tricalcium phosphate was 38.89 mg/L, which is 111.11 times greater than the control treatment (0.35 mg/L), with a significant difference (P < 0.05). The content of soluble potassium in the culture solution is 2.39 mg/L, and the difference with the contrast is obvious.
TABLE 2 determination of phosphate solubilizing Effect of endophytic bacteria by Shake flask method
(5) 16S rDNA sequence analysis
The 16S rDNA gene sequence is shown in a nucleotide sequence table SEQ ID NO. 1. The tested 16S rDNA sequence is compared with the sequence in GenBank database, and the result shows that CT-B09-2 and GenBank databaseEnterobacter aerogenesOn the same branch, the 16S rDNA sequence andEnterobacter aerogenesthe similarity of (JQ 660204) reaches 99%. Combined with colony morphology, physiological and biochemical characteristics and 16S rDNA sequence analysis, is identified as enterobacter aerogenes: (Enterobacter aerogenes）。
(6) Colonization of endophytes
The endophytic bacterium strain CT-B09-2 is subjected to a rifampicin marker method to obtain a stable rifampicin marker resistant mutant strain. In NB medium containing 300. mu.g/ml rifampicin (Rif) at 28 ℃ and 180 r.min-1Shaking culturing for 72 hr, diluting with sterile water to obtain a solution containing 1 × 108cfu/mL of bacterial suspension. Inoculating moso bamboo seedlings by a root irrigation method and a leaf axillary injection method, and respectively taking root, stem and leaf tissues of the moso bamboo seedlings to separate mutant strains 3 d and 7d after inoculation. The test result shows that mutant strains can be recovered from root, stem and leaf tissues of the moso bamboo by the root irrigation method and the axillary injection treatment of the 3 rd and the 7 th inoculation treatment methods, and no bacteria are separated from the control, which shows that the enterobacter aerogenes CT-B09-2 can colonize in the moso bamboo body and can be conducted and propagated.
TABLE 3 colonization and separation results of rhizosphere-irrigation and injection inoculated growth-promoting strains in phyllostachys pubescens roots
Note: + indicates that bacterial colonies can be isolated.
Example 2 potting experiment
The raw moso bamboo is subjected to the endogenous phosphorus and potassium dissolving and nitrogen fixation to aerogenic enterobacter bacteriaEnterobacter aerogenes) CT-B09-2 was activated on the NA slant, and a loop was picked up and inoculated into NA medium (beef extract 3g, peptone 10g, NaCl 5g, agar 18g, water 1000ml, pH 7.0-7.2) at 28 ℃ for 180 r.min-1Shaking culturing for 72 hr, diluting with sterile water to obtain a solution containing 1 × 108cfu/mL of bacterial suspension was used for inoculation. Inoculating the young bamboo seedlings after root irrigation, and performing the seedling culture on the young bamboo seedlings at 15d and 30d by using a portable photosynthesis determinator of LI-6400 type produced by LI-COR corporation of AmericaLeaf net photosynthetic rate, transpiration rate, stomatal conductance and intercellular space CO2Measurement of photosynthesis indexes such as concentration.
After the potted seedling of the moso bamboo is inoculated with endophytic bacteria CT-B09-2 after root irrigation, the photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (Gs) and intercellular CO of the moso bamboo leaf are respectively measured at 15d and 30d2Concentration (Gs). The result shows that the photosynthesis rate (Pn), transpiration rate (Tr) and stomatal conductance (Gs) of the moso bamboo seedling leaves treated by the growth-promoting endophytic bacteria are obviously higher than those of the control, and intercellular CO is2The concentration (Ci) is lower than the control. In particular, the data were most significant at 15 days after the inoculation treatment.
After the potted moso bamboo seedlings are inoculated for 15 days, the leaves of the moso bamboo seedlings are higher than those of the control by 25.66 percent. The blade photosynthetic rate after the growth-promoting endophyte bacterial liquid is inoculated for 30d is higher than that of the clear water control treatment, which is shown in the following table 4.
TABLE 4 Effect of endophytic bacteria CT-B09-2 on the photosynthesis Rate (Pn) of Phyllostachys Pubescens leaves
After the growth-promoting endophytic bacteria are inoculated for 15 days, the transpiration rates of the leaves of the moso bamboo seedlings are higher than those of the control, the transpiration rates are 52.48% higher than those of the control, and the difference between the growth-promoting endophytic bacteria CT-B09-2 and the clear water treatment control is obvious. The leaf transpiration rate after 30 days of inoculation of the growth-promoting endophytic bacteria is higher than that of the clear water control treatment, which is shown in the following table 5.
TABLE 5 Effect of endophytic bacterium CT-B09-2 on the transpiration rate (Tr) of Phyllostachys Pubescens leaves
After the growth-promoting endophytic bacteria are inoculated for 15 days, the stomatal conductance of the leaves of the moso bamboo seedlings is higher than that of a control by 95.45 percent, wherein the difference between the growth-promoting endophytic bacteria CT-B09-2 and a clear water treatment control is obvious. The leaf transpiration rate after 30d inoculation of the growth-promoting endophytic bacteria is higher than that of the clear water control treatment, and the difference is obvious, as shown in the following table 6.
TABLE 6 influence of endophytic bacterium CT-B09-2 on the stomatal conductance (Gs) of Phyllostachys Pubescens leaf
As can be seen from Table 7, after the growth-promoting endophytic bacteria are inoculated for 15 days, the stomatal conductance of the leaves of the phyllostachys pubescens seedlings is lower than that of the control by 22.34 percent, and the difference with that of the clear water treatment control is obvious. Leaf intercellular CO after 30d inoculation of growth-promoting endophytic bacteria2The concentrations were all lower than those of the control treatments with clear water, and the differences were obvious as shown in the following table.
TABLE 7 endophytic bacterium CT-B09-2 vs. intercellular CO of Phyllostachys pubescens leaves2Influence of the concentration (Ci)
Transpiration plays an important role in promoting water transfer and plant absorption, can maintain water saturation of each part of the plant, maintain the form of cell tissues, promote the distribution of inorganic salts in the plant body, and also can dissipate redundant heat energy in photosynthesis and oxidative metabolism of the plant, and meanwhile, the plant must open pores during photosynthesis to obtain required CO from the atmosphere2Therefore, the pro-biotic bacteria can enhance photosynthesis thereof, thereby promoting the growth of moso bamboos.
Example 3 field experiments
Selecting a phyllostachys pubescens forest base of the natural protection area of Longxishan mountain of Happy county of Fujian province, selecting phyllostachys pubescens with the degree of II, firstly drilling a hole at a bamboo pole position which is about 30cm away from the earth surface by using an electric drill, then respectively injecting 30 mL of the bacterial suspension into a bamboo cavity of the phyllostachys pubescens by using a sterile injector, repeatedly inoculating 30 mL of the bacterial suspension on the next day, and sealing the cavity of the bamboo cavity by using soil. Each treatment was repeated 3 times for 10 plants, using clear water as a control. Collecting 5-10 pieces of the bamboo leaves in east, south, west and north directions respectively 15d and 30d after treatment by using a high branch scissors, and fully mixing to obtain a mixed sample for measuring physiological and biochemical indexes such as chlorophyll content, Catalase (CAT), Malondialdehyde (MDA), Peroxidase (POD), superoxide dismutase (SOD), soluble protein content, soluble sugar and the like.
A SPAD-502 chlorophyll rapid determination instrument (Minolta, Japan) is adopted to select 10 phyllostachys pubescens leaves, SPAD values are respectively determined at the leaf base, the leaf and the leaf tip, the average value of each leaf is calculated, and 3 times of repetition are carried out. The relative leaf chlorophyll values are expressed as SPAD values.
Measuring the Catalase (CAT) activity of the moso bamboo leaves by adopting an ultraviolet spectrophotometry method, measuring the Malondialdehyde (MDA) content of the moso bamboo leaves by referring to a thiobarbituric acid method, measuring the Peroxidase (POD) activity of the moso bamboo leaves by referring to a guaiacol method, measuring the superoxide dismutase (SOD) activity of the moso bamboo leaves by referring to a azocyanobenzol (NBT) photoreduction method, measuring the soluble protein content of the moso bamboo leaves by adopting a Coomassie brilliant blue colorimetric method, and measuring the soluble sugar content of the moso bamboo leaves by adopting an Enhomochromatism method.
Influence of growth-promoting endophytic bacteria on chlorophyll content of moso bamboo leaves
As can be seen from figure 1, the SPAD values of the moso bamboo leaves after the injection and inoculation treatment of the endophytic bacteria are higher than those of the control treatment of the clean water inoculation, wherein the SPAD values of the moso bamboo leaves after the inoculation treatment of the endophytic bacteria are obviously different from those of the control treatment of the clean water after the inoculation treatment of the endophytic bacteria CT-B09-2 for 15 days. The chlorophyll content of the moso bamboo leaves treated by the endophytic bacteria is increased, and the chlorophyll plays an important role in light energy absorption, transmission and conversion, so that the photosynthesis rate of the moso bamboo can be improved by increasing the chlorophyll content, and the growth of the moso bamboo is promoted.
Effect of growth-promoting endophytic bacteria on Catalase (CAT) activity of moso bamboo leaves
As can be seen from FIG. 2, the endophytic bacteria treatment has a significant effect on the Catalase (CAT) activity of the moso bamboo leaves, and the catalase activity of the moso bamboo leaves treated by the endophytic bacteria CT-B09-2 is higher than that of the clean water control treatment, and the difference between the catalase activity and the control treatment is significant. After the treatment of endophytic bacteria, the CAT activity in the moso bamboo body can be rapidly increased and maintained at a high level. Catalase (CAT) is an important enzymatic defense system in plants, and can scavenge H2O2Is a plantAn important antioxidant enzyme in vivo.
Effect of growth-promoting endophytic bacteria on Malondialdehyde (MDA) activity of moso bamboo leaves
The results of measuring the change of Malondialdehyde (MDA) concentration of the moso bamboo leaves after inoculation treatment of moso bamboos with II degrees by the endophytic bacteria injection method (figure 3) show that compared with a clear water treatment control, the MDA concentration is kept stable basically or slightly reduced between treatments within 15d and 30d after inoculation of the endophytic bacteria. The MDA content in the host body is related to the damage degree of host cells, and the MDA content increase is a direct factor of plant cell damage, so that the MDA content can be effectively reduced after the moso bamboos are treated by endophytic bacteria, and the effect of protecting the moso bamboo cell membrane is achieved.
Effect of growth-promoting endophytic bacteria on Peroxidase (POD) Activity of Phyllostachys Pubescens leaves
As can be seen from fig. 4, the endophytic bacteria injection treatment had different effects on Peroxidase (POD) activity of the phyllostachys pubescens leaves. The POD activity of the endophytic bacterium CT-B09-2 inoculated for 15d and 30d is obviously higher than that of the clean water control treatment, and the difference is obvious. The endophytic bacteria can improve the Peroxidase (POD) activity of the moso bamboo leaves after being injected and inoculated with the moso bamboos, so that the growth of the moso bamboos is promoted.
Influence of growth promoting endophytic bacteria on superoxide dismutase (SOD) activity of moso bamboo leaf tablet
After endophytic bacterium CT-B09-2 is treated for 15d and 30d, the change of superoxide dismutase (SOD) activity of the moso bamboo leaves is basically similar to the change trend of clear water control treatment, and the trend is stable. The SOD activity of endophytic bacterium CT-B09-2 after inoculation treatment for 15d is obviously higher than that of clear water control treatment, and the difference is obvious (figure 5). Shows that the superoxide dismutase (SOD) activity of the moso bamboo leaves can be improved after the growth promoting strains are inoculated.
Influence of growth-promoting endophytic bacteria on soluble protein content of moso bamboo leaves
After being treated by various endophytic bacteria, the soluble protein content of the moso bamboo leaves and the content of the soluble protein of the clean water control treated moso bamboo leaves show a gradually increasing trend; when the endophytic bacteria are inoculated for 15d and 30d, the content of the soluble protein of the moso bamboo leaves treated by the CT-B09-2 is remarkably different from that of the clean water control (figure 6). Thus, it can be seen that inoculation with endophytic bacteria induces an increase in the soluble protein content in the moso bamboo leaves.
Influence of growth-promoting endophytic bacteria on soluble sugar content of moso bamboo leaves
The measurement results of the soluble sugar content change of the moso bamboo leaves show (fig. 7) that the content of the soluble protein of the moso bamboo leaves treated by various endophytic bacteria and clear water in a contrast manner gradually increases. Compared with a clear water control, the soluble protein content of the moso bamboo leaves is obviously different when the endophytic bacterium CT-B09-2 is inoculated for 15 days.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
SEQUENCE LISTING
<110> Fujian agriculture and forestry university
<120> enterobacter aerogenes for phosphorus and potassium decomposition, nitrogen fixation in phyllostachys pubescens and application thereof
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<160>1
<170>PatentIn version 3.3
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<212>DNA
<213> Artificial sequence
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gggcggtgtg tacaaggccc gggaacgtat tcaccgtggc attctgatcc acgattacta 120
gcgattccga cttcatggag tcgagttgca gactccaatc cggactacga cgcactttat 180
gaggtccgct tgctctcgcg aggtcgcttc tctttgtatg cgccattgta gcacgtgtgt 240
agccctactc gtaagggcca tgatgacttg acgtcatccc caccttcctc cagtttatca 300
ctggcagtct cctttgagtt cccggccgga ccgctggcaa caaaggataa gggttgcgct 360
cgttgcggga cttaacccaa catttcacaa cacgagctga cgacagccat gcagcacctg 420
tctcagagtt cccgaaggca ccaatccatc tctggaaagt tctctggatg tcaagagtag 480
gtaaggttct tcgcgttgca tcgaattaaa ccacatgctc caccgcttgt gcgggccccc 540
gtcaattcat ttgagtttta accttgcggc cgtactcccc aggcggtcga cttaacgcgt 600
tagctccgga agccacgcct caagggcaca acctccaagt cgacatcgtt tacggcgtgg 660
actaccaggg tatctaatcc tgtttgctcc ccacgctttc gcacctgagc gtcagtcttt 720
gtccaggggg ccgccttcgc caccggtatt cctccagatc tctacgcatt tcaccgctac 780
acctggaatt ctacccccct ctacaagact ctagcctgcc agtttcgaat gcagttccca 840
ggttgagccc ggggatttca catccgactt gacagaccgc ctgcgtgcgc tttacgccca 900
gtaattccga ttaacgcttg caccctccgt attaccgcgg ctgctggcac ggagttagcc 960
ggtgcttctt ctgcgggtaa cgtcaattga cgaggttatt aacctcaaca ccttcctccc 1020
cgctgaaagt actttacaac ccgaaggcct tcttcataca cgcggcatgg ctgcatcagg 1080
cttgcgccca ttgtgcaata ttccccactg ctgcctcccg taggagtctg gaccgtgtct 1140
cagttccagt gtggctggtc atcctctcag accagctagg gatcgtcgcc taggtgagcc 1200
gttaccccac ctactagcta atcccatctg ggcacatccg atggcaagag gcccgaaggt 1260
ccccctcttt ggtcttgcga cgttatgcgg tattagctac cgtttccagt agttatcccc 1320
ctccatcagg cagtttccca gacattactc acccgtccgc cactcgtcag cgaagcagca 1380
agctgcttcc tgttaccgtt cgac 1404
Claims (4)
1. A moso bamboo endogenous phosphorus and potassium dissolving nitrogen fixing aerogenic enterobacter strain is characterized in that: the bacterium is enterobacter aerogenes (A)Enterobacter aerogenes) CT-B09-2, already at 2017Is preserved in China general microbiological culture Collection center in 7 months and 3 days in year, and the preservation numbers are: CGMCC No. 14377.
2. A biological agent comprising the Enterobacter aerogenes according to claim 1.
3. The use of enterobacter aerogenes of claim 1 for solubilizing phosphorus, potassium and nitrogen.
4. Use of enterobacter aerogenes according to claim 1 for promoting photosynthesis of phyllostachys pubescens.
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