CN109644818B - Drought-resistant growth-promoting colloidal matrix for forest trees and application thereof - Google Patents
Drought-resistant growth-promoting colloidal matrix for forest trees and application thereof Download PDFInfo
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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Abstract
The invention provides a drought-resistant growth-promoting colloidal matrix for forest trees and a seedling culture matrix applied to the drought-resistant growth-promoting colloidal matrix. The drought-resistant growth-promoting colloidal matrix for the forest trees is formed by uniformly mixing a solid microbial inoculum, grass peat, perlite and vermiculite; wherein the solid microbial inoculum is formed by uniformly inoculating a drought-resistant microorganism combined bacterial liquid into a colloidal fermentation material; the drought-resistant microorganism combined bacterial liquid is prepared by compounding bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei 1.0580. The drought resistance and growth promotion activity of the bacillus subtilis GE3 can be fully exerted, the synergistic symbiosis effect among strains can be exerted in a drought environment, and meanwhile, the mixed fermentation product of the water-washed cow dung and the cow dung biogas residues is added into the substrate, so that the colonization survival performance of microorganisms is further improved, the reutilization of waste resources such as cow dung is realized, and the bacillus subtilis GE3 has good practical application value.
Description
Technical Field
The invention belongs to the technical field of seedling raising substrates and forestry microorganisms, and particularly relates to a drought-resistant growth-promoting colloidal substrate for forest trees and application thereof.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
The substrate is an important component of seedling culture, and is artificial soil prepared by using organic and inorganic materials and microbial agents according to the requirements of seedling growth. Foreign seedling culture substrates mostly adopt a formula of 20 th century sixty-seven years, such as composite substrates A and B of the American cornell university. At present, peat is the best seedling culture medium recognized at home and abroad, and at present, two parts of peat and one part of vermiculite are adopted as the seedling culture medium at home and abroad. The seedling culture substrate has a good seedling culture effect in most plants, but the turf belongs to non-renewable resources, nutrients are easy to lose along with watering of the substrate, the nutrients are difficult to supplement in the later period in a factory mode, and meanwhile, the water requirement during seedling culture is high, so that the great water resource waste is caused.
In recent years, with the rapid development of the scale and intensification of the dairy cow breeding industry, the problem of environmental pollution caused by concentrated excrement discharge in dairy farms is increasingly highlighted. The number of the dairy cattle stored in the nation in 2007 is 1218.9 ten thousand, the daily excretion amount of the dairy manure reaches 20-30 ten thousand t, and particularly in a large-scale dairy farm, if an effective treatment means is lacked, a large amount of dairy manure is accumulated, so that serious pollution is easily caused. Compared with other animal wastes, the total solid content of the cow dung anaerobic fermentation is low in methane yield, the cow dung crude fiber content is high, the percentage of the cow dung crude fiber in dry matter is close to 40%, and the crude fiber degradation is slow. How to efficiently utilize the dung of the cattle farm is one of the difficulties in the breeding industry.
An important purpose of forest seedling raising is to improve the survival rate of afforestation in arid mountainous regions. At present, the improvement of drought tolerance of forest trees by using a microbial technical means becomes a new research topic. However, reports on the preparation of the drought-resistant growth-promoting seedling substrate for the forest trees by combining cattle farm excrement and microbial technologies are not available so far.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the drought-resistant growth-promoting colloid type matrix for forest trees, which is compounded with a plurality of microorganisms with different functional activities, so that the drought-resistant growth-promoting activity of bacillus subtilis GE3 can be fully exerted, the synergistic symbiosis between strains can be exerted in a drought environment, and meanwhile, a mixed fermentation product of water-washed cow dung and cow dung biogas residues is added into the matrix, so that the colonization survival performance of microorganisms is further improved, the reutilization of waste resources such as cow dung and the like is realized, and the matrix has good practical application value.
One of the purposes of the invention is to provide a drought-resistant growth-promoting colloidal matrix for forest trees.
The second purpose of the invention is to provide the application of the drought-resistant growth-promoting colloid matrix for the forest.
In order to achieve the purpose, the invention relates to the following technical scheme:
the invention provides a drought-resistant growth-promoting colloidal matrix for forest trees, which contains Bacillus subtilis GE3, wherein the strain is preserved in China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 Siro No.1 Xin Wen Chen in the sunward area of Beijing city) in 2016 (1 month and 7 days in 2016), and the preservation number of the strain is CGMCC No. 11966.
The Bacillus subtilis GE3 is a plant rhizosphere growth-promoting bacterium suitable for drought stress, the bacterial colony on an LB culture medium is round and irregular, the size of the bacterial colony cultured for 24 hours is about 5mm, the surface is dry, dull and uneven in edge, the bacterial colony is brownish yellow, the middle color is dark, and the bacterial colony is opaque and has spores.
Furthermore, the drought-resistant growth-promoting colloid matrix for the forest trees contains a group of drought-resistant microorganism combination bacteria, and the drought-resistant microorganism combination bacteria are compounded by bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei. Further, the ratio of CFU of bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei is 10: 1-2: 1-2; the CFU ratio of the three strains was determined by the fertilizer efficiency test.
The Bacillus cereus Z77 is preserved in China general microbiological culture Collection center (CGMCC) at 12/31/2012 with the preservation number of CGMCC No.7070 and is described in Chinese patent 'a strain of Bacillus cereus and application thereof in improving plant drought resistance' (CN 103667132A).
The Lactobacillus casei (Lactobacillus casei) is derived from China general microbiological culture Collection center (CGMCC) and can be purchased through a commercial way; further, the lactobacillus casei has a accession number of 1.0580.
Further, the drought-resistant growth-promoting colloid type matrix for the forest trees is prepared by uniformly mixing a solid microbial inoculum, grass peat, perlite and vermiculite.
Wherein the solid microbial inoculum is formed by uniformly inoculating the drought-resistant microorganism combined bacterial liquid into the colloidal fermentation material, and the inoculation proportion is that the colloidal fermentation material: the drought-resistant microorganism combined bacterium liquid composition is 5 kg: 1-2L (W/V);
the drought-resistant microorganism composite bacterial liquid is prepared by compounding bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei 1.0580, and the CFU ratio of the three strains is 10: 1-2: 1-2; the bacterial liquid contains viable bacteria with the number of 6-8 multiplied by 109cfu/ml；
The preparation method of the colloidal fermentation material comprises the steps of uniformly mixing cow dung biogas residues and washed cow dung according to the proportion of 1:1, decomposing the mixed material in a strip stack fermentation mode, wherein the height of a strip stack is more than 1m, the width of the strip stack is more than 3m, the length of the strip stack is more than 20m, turning the stack after the temperature of the stack rises, and allowing the fermentation time to be 4-8 d and air-drying the stack for later use.
Further, the addition mass ratio of the solid microbial inoculum, the grass peat, the perlite and the vermiculite is 20-30: 30-40: 10-15: 5-10.
In a second aspect of the invention, the application of the drought-resistant growth-promoting colloidal matrix for the trees in improving the drought tolerance of the trees and further improving the afforestation survival rate and the preservation rate of arid mountainous regions is provided.
Wherein the forest trees include, but are not limited to, white wax and arborvitae.
Compared with the prior art, the invention has the following effects and advantages:
1. the invention takes the bacillus subtilis GE3 as a main component, optimizes and combines the bacillus cereus Z77 and the lactobacillus casei, realizes the combination of multifunctional strains, fully exerts the synergistic symbiosis among the strains, and can improve the drought tolerance of the forest when being applied in the drought stress.
2. According to the invention, the cow dung biogas residues and the water-washed cow dung are mixed and fermented, so that the reutilization of waste resources is realized, and the fermentation product has certain colloid characteristics, good water absorption and strong water retention, and is suitable for the colonization and survival of microorganisms in the drought-resistant microorganism combined microbial agent.
3. The drought-resistant growth-promoting colloid matrix can save water and reduce diseases during seedling culture, and can improve the water utilization efficiency and the afforestation survival rate and the preservation rate of arid mountainous regions after seedling afforestation.
Drawings
FIG. 1 is a colony morphology of Bacillus subtilis GE3 on LB medium plates;
FIG. 2 is a light mirror image of Bacillus subtilis GE 3;
FIG. 3 is a histogram of the survival rate and preservation rate of the seedlings in the container of the arborvitae of the experimental group and the control group of example 2.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
The present invention is further illustrated by reference to specific examples, which are intended to be illustrative only and not limiting. If the experimental conditions not specified in the examples are specified, they are generally according to the conventional conditions, or according to the conditions recommended by the sales companies; the present invention is not particularly limited, and may be commercially available.
As introduced in the background art, in the prior art, no report on the preparation of the drought-resistant growth-promoting seedling-raising matrix for forest trees by combining cattle farm excrement and microbial technology exists so far.
In view of the above, in a specific embodiment of the present invention, a drought-resistant growth-promoting colloidal matrix for forest trees is provided, wherein the drought-resistant growth-promoting colloidal matrix for forest trees comprises Bacillus subtilis GE3, the strain has been deposited in China general microbiological culture Collection center (CGMCC for short, with the address of CGMCC No. 3, Siro-Lu 1 # 3, Kyoho, Beijing) in 1 month and 7 days of 2016, and the deposition number is CGMCC No. 11966.
The Bacillus subtilis GE3 is a plant rhizosphere growth-promoting bacterium suitable for drought stress, the bacterial colony on an LB culture medium is round and irregular, the size of the bacterial colony cultured for 24 hours is about 5mm, the surface is dry, dull and uneven in edge, the bacterial colony is brownish yellow, the middle color is dark, and the bacterial colony is opaque and has spores.
The physiological and biochemical characteristics are as follows:
the result of the protease determination of the growth promoting bacteria is as follows:
sequencing a 16s rDNA sequence (shown in SEQ ID No.1) of the strain GE3, and identifying the strain GE3 as Bacillus subtilis by combining morphological characteristics and physiological and biochemical characteristics of thalli.
In another embodiment of the invention, the drought-resistant growth-promoting colloidal matrix of the forest trees comprises a group of drought-resistant microorganism combination bacteria, and the drought-resistant microorganism combination bacteria are compounded by bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei. The ratio of CFU of the bacillus subtilis GE3 to the bacillus cereus Z77 to the lactobacillus casei is 10: 1-2: 1-2; the CFU ratio of the three strains was determined by the fertilizer efficiency test.
The Bacillus cereus Z77 is preserved in China general microbiological culture Collection center (CGMCC) at 12/31/2012 with the preservation number of CGMCC No.7070 and is described in Chinese patent 'a strain of Bacillus cereus and application thereof in improving plant drought resistance' (CN 103667132A).
The Lactobacillus casei (Lactobacillus casei) is derived from China general microbiological culture Collection center (CGMCC) and can be purchased through a commercial way;
in yet another embodiment of the present invention, the lactobacillus casei has accession number 1.0580.
In another embodiment of the invention, the drought-resistant growth-promoting colloidal matrix for the forest trees is prepared by uniformly mixing solid microbial inoculum, turf, perlite and vermiculite.
In another embodiment of the present invention, the solid microbial inoculum is prepared by uniformly inoculating the drought-resistant microorganism combined bacterial liquid into the colloidal fermentation material, and the inoculation ratio is that of the colloidal fermentation material: the drought-resistant microorganism combined bacterium liquid composition is 5 kg: 1-2L (W/V);
in another embodiment of the invention, the drought-resistant microbial combined bacterial liquid is prepared by compounding bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei 1.0580, and the CFU ratio of the three strains is 10: 1-2: 1-2; the bacterial liquid contains viable bacteria with the number of 6-8 multiplied by 109cfu/ml；
In another embodiment of the invention, the preparation method of the colloidal fermentation material comprises the steps of uniformly mixing the cow dung biogas residues and the washed cow dung according to the proportion of 1:1, and decomposing the mixed material by adopting a stack fermentation mode to prepare the colloidal fermentation material; specifically, the height of the strip stacks is larger than 1m, the width of the strip stacks is larger than 3m, the length of the strip stacks is larger than 20m, the stacks are turned after the temperature of the stacks rises, the fermentation time is 4-8 d, and the strip stacks are dried for later use.
In another specific embodiment of the invention, the solid microbial inoculum, the grass peat, the perlite and the vermiculite are added in a mass ratio of 20-30: 30-40: 10-15: 5-10.
In another embodiment of the invention, the application of the drought-resistant growth-promoting colloidal matrix for trees in improving the drought tolerance of trees and further improving the afforestation survival rate and the preservation rate of arid mountainous regions is provided.
Wherein the forest trees include, but are not limited to, white wax and arborvitae.
The technical solution of the present invention is further described with reference to the following specific examples. In the embodiment, the lactobacillus casei is derived from China general microbiological culture Collection center (CGMCC), and the preservation number is 1.0580.
Example 1
The substrate is filled in an LKY-1 type non-woven fabric seedling raising forming machine developed by Shandong province forestry scientific research institute. The matrix after molding is cylindrical, 12cm high and 9cm diameter. 5 different test treatments are set, namely inoculation of no microbial inoculum (CK), inoculation of bacillus subtilis DZ1, bacillus subtilis GE3, bacillus cereus Z77, lactobacillus casei and combined bacterial liquid of GE3, Z77 and lactobacillus casei. Wherein the bacillus subtilis DZ1 is preserved in China general microbiological culture Collection center in 2011, 10 months and 14 days, and the preservation number is CGMCC No. 5318. And (5) sowing the seeds after accelerating germination. And (4) setting seedlings in each seedling raising container after 15d after seedling emergence for every 30 seedling raising containers. And measuring the height, the ground diameter and the fresh weight of the white wax seedlings in 15 seedling raising containers after sowing for 90 days. And (5) remaining the nursery stocks, naturally drying, and measuring the relative water content and the relative conductivity of the leaves after 20 days. The results are shown in the table. The combined bacterial liquid can promote the growth of the fraxinus chinensis to different degrees, improve the relative water content of the leaves and reduce the relative conductivity of the leaves, which shows that the combined bacterial liquid has positive effects on promoting the growth of nursery stock and improving the drought resistance of nursery stocks.
TABLE 1 white wax growth and relative water content and relative conductivity of leaves
| Treatment of | Height/cm of seedling | Ground diameter/mm | Total fresh weight/g | Relative water content of leaves (%) | Relative leaf conductivity (%) |
| DZ1 | 68.54±2.54 | 5.13±0.23 | 38.64±1.75 | 76.53±2.53 | 57.35±2.36 |
| GE3 | 71.66±2.42 | 5.94±0.13 | 41.77±1.32 | 80.54±2.66 | 54.26±1.89 |
| Z77 | 69.17±1.96 | 5.44±0.19 | 39.13±1.63 | 76.28±1.85 | 58.63±3.26 |
| Lactobacillus casei | 67.67±2.37 | 5.04±0.20 | 39.04±1.23 | 75.96±1.93 | 56.93±2.14 |
| Combined bacterial liquid | 73.66±2.17 | 6.72±0.17 | 43.37±1.25 | 82.15±2.74 | 52.17±2.81 |
Example 2
The drought-resistant growth-promoting colloid matrix and the conventional container seedling culture matrix are adopted to cultivate the arborvitae container seedlings. The forest establishment test is arranged in the northern mountain of the Niuwangquan of the northern seedling mountain village of the Yangtze towns in the Laiwu city, and the forest establishment test is arranged in 2016, 7 and 12 days. The drought-resistant growth-promoting colloid matrix of the invention and the arborvitae container seedlings cultivated by the conventional container seedling raising matrix are respectively selected for afforestation seedlings. The survival rate was investigated the same year as the forestation, and the survival rate was investigated the next year, and the results are shown in FIG. 3. As can be seen, the afforestation survival rate and the preservation rate of the nursery stock bred by the substrate are obviously higher than those of the conventional seedling substrate.
It should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can modify the technical solution of the present invention as needed or equivalent substitutions without departing from the spirit and scope of the technical solution of the present invention.
SEQUENCE LISTING
<110> scientific research institute of forestry in Shandong province
<120> forest drought-resistant growth-promoting colloid type matrix and application thereof
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1417
<212> DNA
<213> Artificial Synthesis
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Claims (6)
1. The application of the forest drought-resistant growth-promoting colloidal matrix in improving the drought tolerance of trees and further improving the afforestation survival rate and the preservation rate of arid mountainous regions is characterized in that the forest drought-resistant growth-promoting colloidal matrix contains Bacillus subtilis GE3, the strain is preserved in China general microbiological culture Collection center (CGMCC) in 1, 7 and 2016, and the preservation number is CGMCC No. 11966;
the drought-resistant growth-promoting colloidal matrix for the forest trees contains a group of drought-resistant microorganism combination bacteria, and the drought-resistant microorganism combination bacteria are prepared by compounding bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei; the ratio of CFU of the bacillus subtilis GE3 to the bacillus cereus Z77 to the lactobacillus casei is 10: 1-2: 1-2;
the forest is white wax and arborvitae.
2. The application of the drought-resistant growth-promoting colloid matrix for forest trees, which is disclosed by claim 1, in improving the drought tolerance of forest trees and further improving the survival rate and the preservation rate of afforestation in arid mountainous regions, wherein the Bacillus cereus (Z77) is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No. 7070.
3. The application of the drought-resistant growth-promoting colloid type substrate for forest trees, which is disclosed by claim 1, in improving the drought tolerance of the forest trees and further improving the survival rate and the preservation rate of afforestation in arid mountainous regions, wherein the Lactobacillus casei (Lactobacillus casei) is derived from the China general microbiological culture Collection center, and the Lactobacillus casei is deposited with the accession number of 1.0580.
4. The application of the drought-resistant growth-promoting colloidal matrix for the forest in improving the drought tolerance of the forest and further improving the afforestation survival rate and the storage rate of arid mountainous regions is characterized in that the drought-resistant growth-promoting colloidal matrix is prepared by uniformly mixing a solid microbial inoculum, turf, perlite and vermiculite;
wherein the solid microbial inoculum is formed by uniformly inoculating a drought-resistant microorganism combined bacterial liquid into a colloidal fermentation material;
the drought-resistant microorganism combined bacterial liquid is prepared by compounding bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei 1.0580;
the inoculation proportion is that the colloidal type fermentation material: the drought-resistant microorganism combined bacterium liquid composition is 5 kg: 1-2L;
the ratio of CFU of bacillus subtilis GE3, bacillus cereus Z77 and lactobacillus casei 1.0580 was 10: 1-2: 1-2; the bacterial liquid contains viable bacteria with the number of 6-8 multiplied by 109cfu/ml；
The forest is white wax and arborvitae.
5. The application of the drought-resistant growth-promoting colloidal matrix for the trees, which is disclosed by claim 4, in improving the drought tolerance of the trees and further improving the afforestation survival rate and the storage rate of arid mountainous regions, is characterized in that the preparation method of the colloidal fermentation material comprises the steps of uniformly mixing cow dung biogas residues and water-washed cow dung according to the proportion of 1:1, and decomposing the mixed material in a manner of stack fermentation to obtain the colloidal fermentation material.
6. The application of the drought-resistant growth-promoting colloid type matrix for the forest as claimed in claim 4 in improving the drought tolerance of the forest and further improving the afforestation survival rate and the preservation rate of arid mountainous regions, wherein the addition mass ratio of the solid microbial inoculum, the grass peat, the perlite and the vermiculite is 20-30: 30-40: 10-15: 5-10.
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