CN115287210B - Autogenous nitrogen fixation bacteria and application thereof - Google Patents
Autogenous nitrogen fixation bacteria and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
- A01G22/22—Rice
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/40—Fabaceae, e.g. beans or peas
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
Abstract
The invention discloses a strain of autogenous nitrogen fixation bacteria and application thereof. The authigenic nitrogen-fixing bacteria are named Leclercia adecarboxylata QDSM and are preserved in China center for type culture collection, and the preservation number is CCTCC No. M20211653. Research shows that Leclercia adecarboxylata QDSM01 has the capabilities of phosphate dissolution, potassium dissolution and nitrogen fixation, can take sucrose, glucose, fructose and mannitol as carbon sources, and fix nitrogen in air, and has the nitrogen fixation efficiency of fructose, glucose, sucrose and mannitol from high to low respectively; leclercia adecarboxylata QDSM01 can colonize the rhizosphere of corn, soybean and rice, and promote the growth of corn, rice and soybean by promoting the length, number of connections, number of nodes and the synthesis of auxin and gibberellin. The invention provides a new technical means for developing and applying biofertilizer.
Description
Technical Field
The invention relates to a self-growing nitrogen-fixing bacterium and application thereof, in particular to a self-growing nitrogen-fixing bacterium and application thereof in nitrogen fixing efficiency and promoting the growth of corn, soybean and rice when four single carbon sources are utilized. The invention belongs to the technical field of agricultural production.
Background
Nitrogen is an important constituent element of organic molecules and plays an important role in the growth and metabolism of organisms. The nitrogen content is a main influencing factor for measuring the productivity of soil, and plants absorb nitrogen from the soil to maintain vital activities. In order to balance nitrogen in soil and improve productivity, nitrogen fixation is required to be enhanced.
The main modes of nitrogen fixation include natural nitrogen fixation, industrial nitrogen fixation and biological nitrogen fixation, wherein the biological nitrogen fixation mainly utilizes an enzyme system of nitrogen fixation microorganisms to change nitrogen in air into ammonia, compared with artificial nitrogen fixation and natural nitrogen fixation, the biological nitrogen fixation is wider, and the natural world mainly relies on organisms to fix nitrogen, so that the mode plays an important role in nitrogen circulation of an ecological system. Nitrogen-fixing bacteria are classified into three main categories from biological nitrogen-fixing forms: and combining nitrogen-fixing bacteria, symbiotic nitrogen-fixing bacteria and free-living nitrogen-fixing bacteria. The combined azotobacter can not form a specific nitrogen fixation symbiotic structure with host organisms, so that the combined azotobacter is easily influenced by biological stress such as plant diseases and insect pests and abiotic stress such as drought, and the nitrogen fixation efficiency is low. The symbiotic azotobacter has strong host specificity, forms root nodules with specific leguminous plants only, and has single acting object. The free-living nitrogen fixing bacteria have lower nitrogen fixing efficiency, and after the nitrogen source meets the metabolic demand of the free-living nitrogen fixing bacteria, the residual nitrogen source can inhibit the activity of the free-living nitrogen fixing bacteria, but the free-living nitrogen fixing bacteria have wide distribution and strong adaptability, and can survive and fix nitrogen even in soil with deficient nutrition. The free-living nitrogen fixing bacteria can perform catalytic reaction through an in-vivo enzyme system, and the generated nitrogen nutrient substances are supplied to plants for use, so that the crop yield can be remarkably improved. The free-living nitrogen fixing bacteria have the function of producing plant hormone, have important effect on the growth and metabolism of plants, can produce organic acid in addition, and can obviously reduce the content of ineffective potassium in soil, so the free-living nitrogen fixing bacteria are widely used in nitrogen fixing bacteria agents, have wide prospect in application of replacing nitrogen fertilizers, and simultaneously have great influence on agricultural development of China.
The inventor obtains a non-decarboxylated luxuriant bacterium (Leclercia adecarboxylata) by earlier stage separation and purification, named Leclercia adecarboxylata QDSM01, and researches nitrogen fixation efficiency by taking sucrose, glucose, fructose and mannitol as single carbon sources. Corn, soybean and rice are used as test crops, and the promotion effect of the test crops on three crops is studied.
Disclosure of Invention
The invention aims at providing a new isolated autogenous nitrogen fixation bacterium with the capabilities of phosphate dissolution, potassium dissolution and nitrogen fixation.
The second purpose of the invention is to provide the application of the autogenous nitrogen fixation bacteria in promoting the growth of corn, soybean and rice.
In order to achieve the above purpose, the invention adopts the following technical means:
the autogenous nitrogen-fixing bacteria obtained by separation are named Leclercia adecarboxylata QDSM, classified and named Leclercia adecarboxylata QDSM, are preserved in China center for type culture collection (China Center for Type Culture Collection, CCTCC), are at the university of Wuhan, china, and have a preservation number of CCTCC No. M20211653, and have a preservation time of 2021, 12 months and 20 days.
Furthermore, the invention also provides application of the autogenous nitrogen fixation bacteria in promoting the growth of corn, soybean and rice.
Preferably, the autogenous nitrogen fixation bacteria can colonize the rhizosphere of corn, soybean and rice, and can promote the growth of the corn, the rice and the soybean by promoting the length, the connection number, the node number of the roots of the corn, the rice and the soybean and promoting the synthesis of auxin and gibberellin.
Still further, the invention also provides a method for promoting the growth of corn, soybean or rice, which is characterized by comprising the following steps: sterilizing the surfaces of corn, rice or soybean seeds, placing the seeds in an incubator until the seeds germinate to 1-2cm, placing the seeds in fermentation liquor of the authigenic nitrogen-fixing bacteria, and soaking the seeds for 1 hour; then, the seeds were placed in a test tube containing MS solid medium, and the seedling growth was observed for 4-5 days.
Wherein, the concentration of the fermentation liquor of the authigenic nitrogen-fixing bacteria is preferably 1x10 5 、1x10 6 Or 1x10 7 CFU/mL, soaking time was 1 hour.
Compared with the prior art, the invention has the beneficial effects that:
leclercia adecarboxylata genome-wide sequencing results showed that 49 genes were identified in total in the nitrogen metabolism classification, among which 6 nitrosation stresses, 22 nitrate and nitrite ammoniations, 13 ammonia assimilations and 8 denitrification reductase gene clusters. 47 genes are involved in phosphorus metabolism, of which 8 are associated with PHO regulation and high affinity phosphate transporter, phosphate metabolism (22 genes), polyphosphate (3 genes) and alkyl phosphate utilization (14 genes). Finally, there are 5 genes involved in auxin synthesis and 14 genes involved in plant growth promotion in secondary metabolism with higher homology (> 97%) (Snak et al 2021). Therefore, the invention separates and identifies a strain of authigenic nitrogen-fixing bacteria Leclercia adecarboxylata QDSM01, and researches the nitrogen-fixing efficiency when using different carbon sources and the growth-promoting effect of the authigenic nitrogen-fixing bacteria on corns, paddy rice and soybeans. Research shows that Leclercia adecarboxylata QDSM uses sucrose, glucose, fructose and mannitol as carbon sources to fix nitrogen in air, and the nitrogen fixation efficiency is respectively fructose > glucose > sucrose > mannitol from high to low; leclercia adecarboxylata QDSM01 can colonize the rhizosphere of corn, soybean and rice, and promote the growth of corn, rice and soybean by promoting the length, number of connections, number of nodes and the synthesis of auxin and gibberellin. The strain provided by the invention has 22.51% higher IAA secretion than that of the Agrobacterium strain obtained by mutation breeding in the patent application with publication No. CN 102747002A. While the genus Agrobacterium only reports two symbiotic nitrogen fixation gene clusters (nod and nifx) (Guo et al, 2018), 3 of which are involved in phosphorus metabolism (pit, pst and phoU) (Liu et al, 2018). Thus, the strain provided by the present invention has a distinct difference from the Agrobacterium strain, regardless of the strain property study and the partial strain property. Secondly, optimizing a carbon source used by the late fermentation culture medium by the strain, and determining that fructose is a stable carbon source by integrating the nitrogen fixation capacity and long-acting time of the strain; finally, the invention provides a method for promoting the growth of corn, soybean and rice, which has short time for researching the growth promoting effect of bacterial strain and can complete the related research in 5-7 days. The invention provides a high-quality germplasm resource and a simplest and most effective method for promoting the growth of crops.
Drawings
FIG. 1 shows the alignment of QDSM01 strain sequences;
FIG. 2 shows the nitrogen fixation capacity of strain Leclercia adecarboxylata QDSM01 in different carbon sources;
FIGS. 2a-d show the nitrogen fixation capacity of the strain in a medium with sucrose, fructose, glucose and mannitol as single carbon sources, respectively; the different letters in the figures represent significant differences;
FIG. 3 shows the characteristics of strain Leclercia adecarboxylata QDSM 01;
wherein FIGS. 3a-d show the growth of strains on phosphorus-dissolving, potassium-dissolving, nitrogen-fixing and siderophore plates, respectively; FIG. 3e shows IAA and gibberellin secretion of a strain;
FIG. 4 shows the growth promoting effect of strain Leclercia adecarboxylata QDSM01 on the aerial and underground parts of maize seedlings;
wherein, FIGS. 4a-b show the effect of different concentrations of QDSM01 strain on the plant height of corn seedlings, and FIGS. 4c-h show the effect of different concentrations of QDSM01 strain on the fresh weight, root length, root connection number, root node number, root tip number and bifurcation number of the aerial parts of corn seedlings, respectively; the different letters in the figures represent significant differences;
FIG. 5 shows the growth promoting effect of strain Leclercia adecarboxylata QDSM01 on the aerial and underground parts of young rice plants;
wherein, FIGS. 5a-b show the effect of different concentrations of QDSM01 strain on the plant height of rice seedlings, and FIGS. 5c-g show the effect of different concentrations of QDSM01 strain on root length, root connection number, root node number, root tip number and bifurcation number of rice seedlings, respectively; the different letters in the figures represent significant differences;
FIG. 6 shows the growth promoting effect of strain Leclercia adecarboxylata QDSM01 on the aerial and underground parts of soybean seedlings;
wherein, FIGS. 6a-b show the effect of different concentrations of QDSM01 strain on soybean seedling height, and FIGS. 6c-h show the effect of different concentrations of QDSM01 strain on soybean seedling ground fresh weight, root length, root connection number, root node number, root tip number and bifurcation number, respectively; the different letters in the figures represent significant differences.
Detailed Description
The invention is further illustrated by the following experiments in conjunction with examples, which are to be understood as being for illustrative purposes only and in no way limiting the scope of the invention.
Example 1 isolation and identification of the indigenous Nitrogen-fixing bacteria Leclercia adecarboxylata QDSM01
1. Test material
The soil used is the corn rhizosphere soil of the zithohat university Mei Lisi corn straw compost returning field-like field, the collected rhizosphere soil is transported in an ice box, transported back to a laboratory and immediately put into a refrigerator at the temperature of minus 80 ℃ for preservation.
2. Isolation of bacteria
After the rhizosphere soil sample is subjected to gradient dilution, the autogenous nitrogen fixing bacteria are isolated by adopting an Ababetes culture medium. And bacterial 16S RNA sequencing was performed to identify the strain. The strain obtained by separation and purification is named Leclercia adecarboxylata QDSM01 and is preserved in China center for type culture Collection, the address is in university of Wuhan, the microorganism preservation number is CCTCC No. M20211653, and the preservation time is 2021, 12 months and 20 days.
3. Data analysis
Sequencing results of QDSM01 strain Gene BLAST alignment was performed in NCBI (National Center of Biotechnology Information) database and phylogenetic tree was constructed.
4. Results and analysis
The alignment of the QDSM01 strain sequences is shown in FIG. 1. As can be seen from FIG. 1, the isolated strain was most closely related to the genetic evolution of Leclercia adecarboxylata strain TPY2, and the QDSM01 strain was identified as non-decarboxylative luxuriant strain in combination with its morphological and colony characteristics (Leclercia adecarboxylata).
The strain obtained by separation and purification is named Leclercia adecarboxylata QDSM01 and is preserved in China center for type culture Collection, the address is in university of Wuhan, the microorganism preservation number is CCTCC NO. M20211653, and the preservation time is 2021, 12 months and 20 days.
EXAMPLE 2 Leclercia adecarboxylata QDSM01 Nitrogen fixation ability study of Strain
1. Materials and methods
1.1 test strains: leclercia adecarboxylata QDSM01, isolated and identified in example 1.
Beef extract peptone liquid medium: 10.0g of peptone, 5.0g of beef extract, 5.0g of sodium chloride, and water are added to fix the volume to 1L, and the pH value is 7.0.
Basic culture medium of albe: 0.2g of monopotassium phosphate, 0.2g of magnesium sulfate, 0.2g of sodium chloride, 5g of calcium carbonate and 0.1g of calcium sulfate, and adding water to fix the volume to 1L. 10g of sucrose, 10g of fructose, 10g of glucose and 10g of mannitol are respectively weighed as carbon sources and added into a basic culture medium.
1.2 Leclercia adecarboxylata QDSM01 Nitrogen fixation Capacity detection
100 mu L of bacterial liquid is inoculated into 100mL of beef extract peptone liquid culture medium and cultured for 24 hours at 30 ℃ and 180 r/min. 100 mu L of the seed solution is inoculated into 100mL of Abbe's liquid culture medium with different carbon sources, the culture is carried out at 30 ℃ and 180r/min, sampling is carried out every 48h, and the nitrogen content in the culture medium is measured by adopting a carbon-nitrogen analyzer liquid module.
1.3 data processing
All raw data of this experiment were initially collated using Microsoft Excel Office 2016 software. The experimental results were plotted using R.
2. Results and analysis
The nitrogen fixation capacity of the Leclercia adecarboxylata QDSM01 strain in different carbon sources is shown in fig. 2. FIG. 2a shows that in a medium with sucrose as a single carbon source, the nitrogen fixation amount of the Leclercia adecarboxylata QDSM strain is highest at 144h, then 48h and 96h, and the nitrogen fixation amount is remarkably reduced at 192h, probably because the nitrogen sources in the medium are more, the growth of the strain is inhibited, and the nitrogen fixation capacity of the strain cannot be effectively expressed; FIG. 2b shows that strain Leclercia adecarboxylata QDSM01 had the highest nitrogen fixation at 192h, followed by 144h, 96h and 48h when fructose was the sole carbon source; FIG. 2c shows that the Leclercia adecarboxylata QDSM strain has the highest nitrogen fixation amount at 48-144 h and 192h next when glucose is used as a single carbon source; FIG. 2d shows that the Leclercia adecarboxylata QDSM strain had the highest nitrogen fixation at 144h in a medium with mannitol as the sole carbon source, compared to 0 h.
By combining the results, the Leclercia adecarboxylata QDSM01 strain can fix nitrogen in air by taking sucrose, glucose, fructose and mannitol as carbon sources, and the nitrogen fixation efficiency is respectively fructose, glucose, sucrose and mannitol from high to low.
Example 2 Leclercia adecarboxylata QDSM01 Strain characterization and growth-promoting related Property analysis
1. Materials and methods
1.1 test materials
1.1.1 test strains
Leclercia adecarboxylata QDSM01, isolated and identified in example 1.
1.1.2 Medium
Ababetes medium: 0.2g of monopotassium phosphate, 0.2g of magnesium sulfate, 0.2g of sodium chloride, 5g of calcium carbonate, 0.1g of calcium sulfate, 10g of mannitol and 20.00g of agar, and adding water to fix the volume to 1L.
Phosphate-dissolving solid medium: 10.00g of glucose, 0.30g of magnesium sulfate heptahydrate, 0.50g of ammonium sulfate, 0.30g of sodium chloride, 2.00g of calcium phosphate, 0.036g of iron sulfate hepta Shui Ya, 0.30g of potassium chloride, 0.03g of manganese sulfate, 20.00g of agar and water are added to fix the volume to 1L.
Potassium-dissolving solid culture medium: sucrose 0.50g, calcium carbonate 0.10g, disodium hydrogen phosphate 0.50g, ferric trichloride 0.05g, magnesium sulfate 0.50g, potassium feldspar 0.10g, agar 20.00g, and water to a volume of 1L.
Beef extract peptone liquid medium: 10.0g of peptone, 5.0g of beef extract, 5.0g of sodium chloride, and water are added to fix the volume to 1L, and the pH value is 7.0.
1.1.3 test seeds
The corn, soybean and rice seeds to be tested are the reclaimed Jiujiu, zhonghuang Jiujiu and Longjing 46 respectively.
1.2 Leclercia adecarboxylata QDSM01 Nitrogen fixation Capacity detection
100. Mu.L of QDSM01 strain is inoculated into 150mL of beef extract peptone liquid medium, and cultured for 24 hours at 30 ℃ and 180r/min to obtain QDSM01 seed liquid. The 3. Mu.L seed solution was inoculated into solid Abbe's disease culture medium, 3 parallel strains were prepared, and after incubation at 30℃for 48 hours, the strains were observed for growth.
1.3 Leclercia adecarboxylata QDSM01 potassium-decomposing ability detection
After inoculating the above 3. Mu.L of QDSM01 seed solution into a potassium-dissolving solid medium and culturing for 48 hours at 30℃in parallel, it was observed whether the strain could grow (Rajawat et al, 2016, wang et al, 2018).
1.4 Leclercia adecarboxylata QDSM01 phosphorus dissolving capacity detection
The seed solution of 3. Mu.L QDSM01 described above was inoculated into phosphorus-soluble solid medium (Wang et al, 2018, amaya-G. Mu.z et al, 2020) and 3 replicates were performed, and after incubation at 30℃for 48 hours, the strain was observed for growth.
1.5 Leclercia adecarboxylata QDSM01 growth hormone (IAA) secretion assay
IAA standards at 200. Mu.g/mL were diluted in gradient to concentrations of 0, 25, 50, 75, 100, 125, 150 and 175. Mu.g/mL and absorbance was measured using Salkowski chromogenic assay (Ahmad et al, 2008). The IAA standard curve equation is determined to be y=0.0055x-0.0002, R 2 ＝0.9967。
The seed solution of QDSM01 was inoculated into a liquid beef extract peptone medium containing 200mg/L tryptophan at an inoculum size of 1%, shake-cultured at 30℃for 120r/min, and IAA was measured every 24 hours by Salkowski chromogenic method.
1.6 Leclercia adecarboxylata QDSM01 siderophore Capacity assay
MKB medium (1L): (1) 5.0g of casein amino acid, 15.0mL of glycerol and 20g of agar powder; (2) 2.5g of K2HPO4; (3) 2.5g MgSO4.7H2O. The three media were dissolved in ultrapure water in a total volume of 1000mL. Mixing the sterilized culture medium.
CAS dye liquor: a0.079 g CAS (chrome azure), dissolved in 50mL deionized water, added with 10 mL1mmol/L Fecl 3 A solution; 0.069g of cetyltrimethylammonium bromide was dissolved in 40mL of deionized water, solution A was slowly added to B along the cup wall, mixed well, and passed through 0.45 μm in an ultra clean bench. 50mL dye was added to the medium and mixed well to prepare CAS plates (Amaya-Gmz et al 2020).
After inoculating the seed solution of 3. Mu.L QDSM01 described above into a CAS solid medium and incubating for 48 hours at 30℃in 3 replicates, it was observed whether orange-yellow halos formed, and if so, siderophores were detected as positive.
1.7 Leclercia adecarboxylata QDSM01 gibberellin secretion assay
Gibberellin is dissolved in 70% ethanol to prepare 100. Mu.g/mL gibberellin standard solution, which is diluted to 0, 10, 30, 40, 50 and 60. Mu.g/mL in a gradient, 5mL of gibberellin solution of each concentration is mixed with 4.5mL of 98% sulfuric acid, and the absorbance at 412nm is measured (Xiao et al, 1997). Gibberellin standard curve equation y=0.0029x-0.0012, R is obtained through measurement 2 ＝0.9993。
The seed solution of QDSM01 was inoculated into a beef extract peptone medium at 1% of the inoculum size, and cultured at 30℃for 48 hours at 180r/min to determine gibberellin concentration of the strain.
1.8 Leclercia adecarboxylata QDSM01 measurement of the ability of maize, soybean and Rice to promote growth
Selecting full corn, soybean and rice seeds without worm holes, sterilizing the seeds with 5% sodium hypochlorite and 75% alcohol for 5min, and cleaning with sterile water for 3 times each for 1min. The seeds were placed in a sterilization plate with filter paper placed up and down and soaked with sterile water. Seeds were placed in an incubator and light (16/8 hour light/dark cycle), temperature (22.+ -. 2 ℃) and relative humidity (60% -70%) were controlled. Germinating the seeds to 1-2cm, soaking the seeds in 0, 1x10 5 、1x10 6 And 1x10 7 Soaking in CFU/mL bacterial liquid for 1 hr, and planting the seeds in MS solid culture mediumIn the tube. Placing the test tube in an incubator, culturing for 4-5 days, measuring plant height and fresh weight, and analyzing the root system shape by a root system scanner.
1.9 data processing
Preliminary finishing was performed using Microsoft Excel Office 2016 software. Drawing was performed using R.
2. Results and analysis
2.1 Leclercia adecarboxylata QDSM01 Strain characterization
And (3) respectively inoculating Leclercia adecarboxylata QDSM strain to nitrogen fixation, potassium dissolution, phosphorus dissolution and siderophore production screening culture mediums for culturing and observing the growth condition of the strain, and simultaneously detecting IAA and gibberellin secretion amounts. The results of the Leclercia adecarboxylata QDSM01 strain characterization are shown in FIGS. 3a-e, which were grown on the above medium and produced halos, and which were determined to have phosphate-solubilizing (FIG. 3 a), potassium-solubilizing (FIG. 3 b), nitrogen-fixing (FIG. 3 c) and iron-producing carriers (FIG. 3 d). FIG. 3e shows that IAA and gibberellin secretion amounts of the strain are 46.31-50.32. Mu.g/mL and 41.10-59.72. Mu.g/mL, respectively.
2.2 Protoffee of Leclercia adecarboxylata QDSM01 strain on corn
The growth promoting effect of the Leclercia adecarboxylata QDSM01 strain on the aerial and underground parts of maize seedlings is shown in FIG. 4. FIGS. 4a-b show the effect of different concentrations of Leclercia adecarboxylata QDSM01 strain on maize seedling height, 10 of which 5 And 10 6 The height of corn seedlings after cfu/mL Leclercia adecarboxylata QDSM01 strain treatment is obviously higher than that of the control group and 10 7 cfu/mL; FIG. 4b shows, 10 5 The fresh weight of the overground parts of the corn seedlings after the cfu/mL Leclercia adecarboxylata QDSM01 strain treatment is obviously higher than that of the control group and 10 7 cfu/mL of 10 6 cfu/mL with control group and 10 5 There was no significant difference in fresh weight of corn seedlings between cfu/mL treatments, while 10 7 There was no significant difference in fresh weight of corn seedlings between cfu/mL and control group; FIGS. 4a and 4d-h,10 6 And 10 7 The root length, root connection number, root node number, root tip number and bifurcation number of the corn seedlings after the cfu/mL Leclercia adecarboxylata QDSM strain treatment are obviously higher than those of the control groupAnd 10 5 cfu/mL, while other root index 10 besides root length 5 cfu/mL was significantly higher than control. To sum up 10 6 The cfu/mL Leclercia adecarboxylata QDSM01 strain is best for maize pro-effect.
2.3 Protoffee of Leclercia adecarboxylata QDSM01 strain on rice
The growth promoting effect of Leclercia adecarboxylata QDSM01 strain on the aerial and underground parts of rice seedlings is shown in FIG. 5. FIGS. 5a-b show the effect of different concentrations of strain Leclercia adecarboxylata QDSM01 on the plant height of young rice seedlings, 10 of these 5 And 10 6 Rice seedling height after cfu/mL Leclercia adecarboxylata QDSM01 strain treatment and control group and 10 7 No significant difference in cfu/mL, 10 at the same time 7 The height of the rice seedlings after cfu/mL Leclercia adecarboxylata QDSM01 strain treatment is obviously higher than that of a control group; FIGS. 5c-d show, 10 5 The root length and root connection number of the rice seedlings after the cfu/mL Leclercia adecarboxylata QDSM01 strain treatment are obviously higher than 10 6 And 10 7 cfu/mL, next is control group; FIG. 5e shows that the number of root nodes of the rice is 10 from high to low 5 cfu/mL>10 7 cfu/mL>Control, at the same time 10 6 cfu/mL and 10 5 cfu/mL and 10 7 There was no significant difference in cfu/mL; FIG. 5f shows that the number of root tips of the rice is 10 from high to low 6 And 10 7 cfu/mL>10 5 cfu/mL>A control; FIG. 5g shows that the number of branches of rice is 10 from high to low 6 cfu/mL> 10 5 cfu/mL>Control, at the same time 10 7 cfu/mL and 10 5 cfu/mL and 10 6 There was no significant difference in cfu/mL; to sum up 10 5 The cfu/mL Leclercia adecarboxylata QDSM01 strain is best for maize pro-effect.
2.4 Protoffee of Leclercia adecarboxylata QDSM01 strain on soybean
The growth promoting effect of Leclercia adecarboxylata QDSM01 strain on the aerial and underground parts of soybean seedlings is shown in FIG. 6. FIGS. 6a-c and 6g show that different concentrations of strain Leclercia adecarboxylata QDSM01 have no effect on soybean seedling height, fresh weight of aerial parts and root tip number; FIG. 6d shows soybeanThe root length of the seedling is respectively 10 from high to low 7 cfu/mL>10 6 And 105cfu/mL>A control; FIGS. 6e-f show that the number of soybean root connections and nodes are 10 from high to low, respectively 7 cfu/mL>10 5 cfu/mL>Control, of which 10 6 cfu/mL and 10 5 cfu/mL and control were not significantly different; FIG. 6h shows that the number of branches of soybean is 10 from high to low 7 cfu/mL>10 5 cfu/mL>10 6 cfu/mL>A control; to sum up 10 7 The cfu/mL Leclercia adecarboxylata QDSM01 strain is best for soybean root promotion effect.
Conclusion:
leclercia adecarboxylata QDSM01 the nitrogen fixation efficiency is fructose, glucose, sucrose and mannitol from high to low respectively; leclercia adecarboxylata QDSM01 can colonize the rhizosphere of corn, soybean and rice, and promote the growth of corn, rice and soybean by promoting the length, number of connections, number of nodes and the synthesis of auxin and gibberellin.
Claims (5)
1. The authigenic nitrogen-fixing bacteria are named Leclercia adecarboxylata QDSM, are preserved in China center for type culture collection (China Center for Type Culture Collection, CCTCC), and have a preservation number of CCTCC No: m2021165.
2. Use of the authigenic nitrogen-fixing bacterium of claim 1 to promote the growth of corn, soybean and rice.
3. The use of claim 2 wherein the indigenous nitrogen fixing bacteria are capable of colonizing corn, soybean and rice rhizosphere and promoting the growth of corn, rice and soybean by promoting the length, number of links, node number and synthesis of auxin and gibberellin.
4. A method for promoting the growth of corn, soybean or rice comprising the steps of: sterilizing the surface of corn, rice or soybean seeds, placing the seeds in an incubator until the seeds germinate to 1-2cm, placing the seeds in the fermentation broth of the authigenic nitrogen-fixing bacteria in claim 1, and soaking the seeds for 1 hour; then, the seeds were placed in a test tube containing MS solid medium, and the seedling growth was observed for 4-5 days.
5. The method of claim 4, wherein the concentration of the fermentation broth of the indigenous nitrogen fixing bacteria is 1x10 5 、1x10 6 Or 1x10 7 CFU/mL, soaking time was 1 hour.
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