CN115812719B - Compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan and application of compound composition in promoting cucumber seedling growth - Google Patents
Compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan and application of compound composition in promoting cucumber seedling growth Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
A compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan and application thereof in promoting cucumber seedling growth belong to the technical field of agricultural planting, the compound composition is in a liquid form, wherein the concentration of the chitosan oligosaccharide is 100-700mg/L, and the viable count of the bacillus amyloliquefaciens is 10 5 ‑10 8 cfu/mL, wherein the lentinan is 400-1000 times of lentinan diluent. The compound composition can effectively promote growth vigor, root system development and photosynthesis of cucumber seedlings, has a promoting effect obviously greater than that of independently applying a chitosan oligosaccharide solution or a bacillus amyloliquefaciens culture solution or a lentinan solution, is also superior to that of compounding the chitosan oligosaccharide solution and the bacillus amyloliquefaciens, and shows obvious synergistic effect. Meanwhile, the compound composition has more remarkable effect on improving matrix soil nutrients, can remarkably relieve the damage of low-temperature stress to cucumber seedling growth vigor, and improves the low-temperature resistance of cucumber seedlings.
Description
Technical Field
The invention belongs to the technical field of agricultural planting, and particularly relates to a compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan and application thereof in promoting cucumber seedling growth.
Background
Biological source substances act on plants, and the plant has obvious stress resistance and growth promotion effects and is harmless to the environment and agricultural products. Studies have shown that biological substances such as biostimulants, plant immunity elicitors and biological bactericides are particularly remarkable in capability of improving stress resistance and crop quality of crops.
Cucumber is one of the main vegetables planted in China, the cucumber planting area is greatly improved in recent years, and pollution and harm are caused to the environment and the cucumber quality due to the fact that chemical fertilizers and pesticides are applied in large quantities to improve the cucumber yield, so that the cucumber quality is obviously reduced, and meanwhile, the cucumber growth is in a sub-health state, so that sustainable development of the cucumber industry is not facilitated. The existing single biological source substance has limited promotion effect on cucumber growth, so that research and development of the composite biological source preparation capable of remarkably improving cucumber quality has important significance for sustainable development of cucumber facility cultivation industry.
Disclosure of Invention
In view of the above, the invention aims to provide a compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan, which can effectively promote the growth of cucumber seedlings, and provides a new theoretical basis and technical means for improving the comprehensive production capacity of cucumbers.
The technical scheme adopted by the invention is as follows:
the invention provides a compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan, wherein the compound composition is in a liquid form, the concentration of the chitosan oligosaccharide is 100-700mg/L, and the viable count of the bacillus amyloliquefaciens is 10 5 -10 8 cfu/mL, wherein the lentinan is 400-1000 times of lentinan diluent.
Preferably, the concentration of the chitosan oligosaccharide is 500mg/L, and the viable count of the bacillus amyloliquefaciens is 10 7 cfu/mL, the lentinan is 700 times lentinan diluent.
The invention also provides application of the compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan in promoting cucumber seedling growth.
The beneficial effects are that: the compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan provided by the invention can effectively promote cucumber seedling growth, root system development and photosynthesis. The field test shows that the promoting effect of the compound composition is obviously greater than that of independently applying chitosan oligosaccharide solution or bacillus amyloliquefaciens culture solution or lentinan solution, and is better than that of the compound composition of chitosan oligosaccharide solution and bacillus amyloliquefaciens culture solution, and the compound composition shows obvious synergistic effect. The analysis of the combination test results shows that the reason that the compound composition has beneficial growth promoting effect is as follows: the chitosan oligosaccharide in the compound composition can enhance the activity index of plant cells, promote the growth of root systems and stems and leaves of crops, improve the chlorophyll content and improve the photosynthesis efficiency; the lentinan can promote the improvement of the plant height, stem thickness and chlorophyll content of cucumber seedlings; the interaction exists between the bacillus amyloliquefaciens and the chitosan oligosaccharide, the bacillus amyloliquefaciens can secrete the chitosan enzyme to form the chitosan oligosaccharide with higher biological activity so as to enhance the biological activity of the chitosan oligosaccharide, and the chitosan oligosaccharide can promote the proliferation of the bacillus amyloliquefaciens, so that the bacillus amyloliquefaciens is beneficial to colonizing the root system of cucumber seedlings, and the activation of matrix nutrients and the root system development of the cucumber are promoted.
In addition, the compound composition of the chitosan oligosaccharide, the bacillus amyloliquefaciens and the lentinan has more remarkable effect on improving the matrix soil nutrients; and the root irrigation treatment of the compound composition obviously relieves the damage of low-temperature stress to the growth vigor of cucumber seedlings, improves the low-temperature resistance of the cucumber seedlings, and has important significance for cultivating strong cucumber seedlings and ensuring the safe overwintering of the cucumber seedlings.
Drawings
FIG. 1 shows the effect of treatment with different concentrations of chitosan oligosaccharide on cucumber seedling vigor.
FIG. 2 shows the effect of treatment with different concentrations of chitosan oligosaccharide on accumulation of cucumber seedling biomass.
FIG. 3 is the effect of the treatment of the compounded composition on cucumber seedling growth under low temperature stress.
FIG. 4 is the effect of the treatment of the compounded composition on the biomass accumulation of cucumber seedlings under low temperature stress.
FIG. 5 is the effect of the treatment of the compounded composition on the antioxidant enzyme activity of cucumber seedlings under low temperature stress.
FIG. 6 is the effect of the treatment of the compounded composition on malondialdehyde content of cucumber seedlings under low temperature stress.
FIG. 7 is a graph showing the growth of Bacillus amyloliquefaciens XY-13 under chitosan oligosaccharide.
Detailed Description
The technical scheme and technical effects of the embodiments of the present invention are further described in detail below with reference to the accompanying drawings.
The test methods used in the following examples are conventional methods unless otherwise specified; the sources of the materials for this test are as follows:
the lentinan is purchased from Beijing Sanpu hundred grass green plant preparation limited company and is water aqua with the active ingredient content of 1 percent.
The chitosan oligosaccharide is provided by the university of Heilongjiang eight-agricultural reclamation.
Bacillus amyloliquefaciens is Bacillus amyloliquefaciens XY-13 (CCTCC M2021270), and is provided by a university of Ningxia microbiological laboratory.
The invention provides a compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan, wherein the compound composition is in a liquid form, the concentration of the chitosan oligosaccharide is 100-700mg/L, and the viable count of the bacillus amyloliquefaciens is 10 5 -10 8 cfu/mL, wherein the lentinan is 400-1000 times of lentinan diluent.
Preferably, the concentration of the chitosan oligosaccharide is 500mg/L, and the viable count of the bacillus amyloliquefaciens is 10 7 cfu/mL, the lentinan is 700 times lentinan diluent.
The invention also provides application of the compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan in promoting cucumber seedling growth.
The preparation method of the compound composition is that the bacillus amyloliquefaciens culture solution with the preset thallus concentration is used as mother liquor, and the preset amounts of chitosan oligosaccharide and lentinan are respectively added into the mother liquor, so that the concentration of the chitosan oligosaccharide and the lentinan in the compound system reaches a preset value. Taking 1L of compound composition liquid as an example, 0.5g of chitosan oligosaccharide and 1.42ml of lentinan are dissolved in 1L10 7 In cfu/mL bacillus amyloliquefaciens XY-13 bacterial liquid, 500mg/L chitosan oligosaccharide +10 is prepared 7 cfu/mL of bacillus amyloliquefaciens XY-13+700 times of lentinan mixed solution; when preparing single chitosan oligosaccharide or lentinan diluent, 1L of clear water is used for dissolution; when two kinds of compound combinations are prepared, if a bacterial agent exists, 1L of bacterial liquid is dissolved.
The growth promoting effect of the compound composition of the invention on cucumber seedlings will be described through specific experiments.
Example 1 test of growth promotion of cucumber seedlings by Chitosan oligosaccharide
In the test, one-leaf Yixindell LD-1 cucumber seedlings are taken as test materials, additional fertilizer is not required during the seedling raising period, irrigation is uniformly carried out according to each treatment of the crop growth requirement in the seedling raising period, and 1 liter of water is irrigated into one plug (72 holes) at a time. The chitosan oligosaccharide concentration is set to be 100mg/L, 200mg/L, 300mg/L, 400mg/L, 500mg/L, 600mg/L and 700mg/L, the water irrigation is used as a control, 3 root irrigation treatments are respectively carried out on the 1 st day, the 10 th day and the 20 th day of the treatment, 8 treatments are carried out in total, each treatment is repeated for 3 times, 40 plants are irrigated for each treatment, 15mL are irrigated for each plant, all treatments are used for unified water management, and seedling growth indexes are counted.
Treatment 1: blank control, water only
Treatment 2: chitosan oligosaccharide solution with irrigation concentration of 100mg/L
Treatment 3: chitosan oligosaccharide solution with irrigation concentration of 200mg/L
Treatment 4: chitosan oligosaccharide solution with irrigation concentration of 300mg/L
Treatment 5: chitosan oligosaccharide solution with irrigation concentration of 400mg/L
Treatment 6: chitosan oligosaccharide solution with irrigation concentration of 500mg/L
Treatment 7: chitosan oligosaccharide solution with irrigation concentration of 600mg/L
Treatment 8: chitosan oligosaccharide solution with irrigation concentration of 700mg/L
As shown in fig. 1, the relative plant height growth rate, the relative stem growth rate, the chlorophyll content and the strong seedling index of the chitosan oligosaccharide treatment are all remarkably higher than those of the blank Control (CK) treatment, and the chitosan oligosaccharide treatment shows a tendency of increasing and then decreasing with the increase of the chitosan oligosaccharide concentration, wherein the treatment performance is optimal when the chitosan oligosaccharide solution concentration is 500mg/L (K500). Compared with CK treatment, the relative growth rate of the treated plant height, the relative growth rate of the stem thickness, the chlorophyll content and the strong seedling index are obviously improved by 39.5%, 87.9%, 17.8% and 70.8% when the concentration of the chitosan oligosaccharide solution is 500 mg/L.
As shown in FIG. 2, the fresh weight of the aerial parts treated by the chitosan oligosaccharide is obviously higher than that of the aerial parts treated by the blank control, and the dry weight of the aerial parts treated by the chitosan oligosaccharide with the concentration of 400mg/L, 500mg/L and 600mg/L (K400, K500 and K600) is obviously higher than that of the aerial parts treated by the blank Control (CK). Wherein the dry and fresh weight of the overground part treated by K500 is optimal, and the dry and fresh weight of the underground part is improved by 43.2%, 70.8%, 77.1% and 105.9% relative to CK. By combining various growth indexes, 500mg/L chitosan oligosaccharide solution can obviously improve cucumber seedling growth vigor and promote biomass accumulation.
TABLE 1 Effect of treatment with Chitosan oligosaccharide at different concentrations on substrate chemistry
As can be seen from table 1, as the concentration of chitosan oligosaccharide increases, the nutrient content of the substrate tends to increase and decrease. The content of quick-acting nitrogen, quick-acting potassium and total nitrogen in the 500mg/L chitosan oligosaccharide solution is highest compared with other treatments, and the content of quick-acting nitrogen and total nitrogen is obviously higher than 156.9% and 20% of that in the blank control treatment. There were no significant differences between treatments for other indicators. In the comprehensive view, the improvement effect of the 500mg/L chitosan oligosaccharide solution root irrigation on the nutrients is optimal.
TABLE 2 Effect of different concentrations of Chitosan oligosaccharide treatments on growth characteristics of cucumber seedling root systems
As can be seen from table 2, the root length, root surface area, and root volume of the chitosan oligosaccharide treatment were significantly higher than those of the blank treatment, and the tendency of increasing and decreasing with increasing chitosan oligosaccharide concentration was shown. Wherein, the 500mg/L chitosan oligosaccharide solution treatment performs optimally, which is significantly higher than other treatments, and the root diameters of the treatments have no significant difference. Compared with the blank control treatment, the root length, the root surface area and the root volume of the 500mg/L chitosan oligosaccharide solution treatment are obviously improved by 59.1 percent, 106.1 percent and 134.3 percent.
TABLE 3 Effect of different concentrations of Chitosan oligosaccharide treatment on photosynthetic Properties of cucumber seedlings
As can be seen from table 3, there was no significant difference in transpiration rate between treatments. The net photosynthetic rate and stomatal conductance of chitosan oligosaccharide treatment are both significantly higher than those of blank control treatment, and the intercellular CO 2 The concentrations were all higher than the blank. Net photosynthetic rate of 500mg/L chitosan oligosaccharide solution treatment, intercellular CO 2 The concentration and the air pore conductivity are optimal, and the concentration and the air pore conductivity are obviously improved by 121.0%, 15.6% and 21.1% compared with the blank control treatment.
Example 2 growth-promoting test of cucumber seedlings by Bacillus amyloliquefaciens and Chitosan oligosaccharide combination
The test was performed with 500mg/L chitosan oligosaccharide in combination with Bacillus amyloliquefaciens. The concentration of the bacillus amyloliquefaciens is set to 10 5 cfu/mL、10 6 cfu/ml、10 7 cfu/ml、10 8 cfu/ml, bacillus amyloliquefaciens selected in the laboratory effectively promotes the growth of cucumber seedlings to obtain bacillus amyloliquefaciens XY-13. Taking pure water root irrigation as control, respectively performing root irrigation treatment for 3 times on 1 st, 10 th and 20 th days, and repeating each treatment for 3 timesEach treatment was carried out for 40 plants, each plant was filled with 15ml of liquid, and seedling growth was measured on day 30.
Treatment 1: blank control, water only
Treatment 2:500mg/L chitosan oligosaccharide solution
Treatment 3:10 7 cfu/ml XY-13 culture solution
Treatment 4:500mg/L chitosan oligosaccharide solution +10 5 cfu/ml XY-13 culture solution
Treatment 5:500mg/L chitosan oligosaccharide solution +10 6 cfu/ml XY-13 culture solution
Treatment 6:500mg/L chitosan oligosaccharide solution +10 7 cfu/ml XY-13 culture solution
Treatment 7:500mg/L chitosan oligosaccharide solution +10 8 cfu/ml XY-13 culture solution
TABLE 4 influence of Bacillus amyloliquefaciens and chitooligosaccharides on cucumber seedling growth
As shown in table 4, each growth condition index of the chitosan oligosaccharide and bacillus amyloliquefaciens compound treatment is obviously higher than that of the blank control treatment, and meanwhile, the relative growth rate of the stem thickness, the chlorophyll content and the strong seedling index are obviously higher than those of the chitosan oligosaccharide alone; the relative growth rate of the stem thickness and the strong seedling index are obviously higher than those of the bacillus amyloliquefaciens alone. Treatment in compounding treatment 6 (500 mg/L Chitosan oligosaccharide+10) 7 cfu/ml XY-13), the plant height relative growth rate, the stem thickness relative growth rate and the chlorophyll content are highest and are obviously higher than those of blank control treatment by 50%, 100% and 53.8%.
TABLE 5 influence of Bacillus amyloliquefaciens and chitooligosaccharides on cucumber seedling root development
As can be seen from table 5, the root development index of each compound treatment was significantly improved compared to the blank control treatment. The root volume and root diameter of each compound treatment are obviously higher than those of the chitosan oligosaccharide treatment and the bacillus amyloliquefaciens treatment. Treatment 6 (500 mg/L Chitosan oligosaccharide+10) in the compounding treatment relative to treatment 1 7 cfu/ml XY-13) root volume, root diameter was 261.4%, 53.3% higher.
TABLE 6 influence of Bacillus amyloliquefaciens and chitooligosaccharides on photosynthesis of cucumber seedlings
As can be seen from table 6, the transpiration rate, net photosynthetic rate, and stomatal conductance of each of the compounding treatments were significantly higher than that of treatment 1. Treatment in compounding treatment 6 (500 mg/L Chitosan oligosaccharide+10) 7 The transpiration rate, net photosynthetic rate of cfu/ml XY-13) performed best, being 227.3%, 134.0% higher relative to treatment 1. The pore conductance of treatment 7 was highest, 300.0% higher than that of treatment 1.
TABLE 7 influence of different treatments on the substrate chemistry
As can be seen from Table 7, the compound combination of chitosan oligosaccharide and Bacillus amyloliquefaciens all increased the chemical content of the matrix except for the quick-acting phosphorus relative to treatment 1. The organic matter and quick-acting nitrogen content of the compound combination are obviously higher than that of the blank control treatment. Wherein, treatment 6 (500 mg/L chitosan oligosaccharide+10) 7 cfu/ml XY-13) the content of EC, organic matter, quick-acting nitrogen, quick-acting potassium and total nitrogen is obviously higher than that of other treatments, and compared with treatment 1, the content of the organic matter, quick-acting nitrogen and total nitrogen is obviously higher than 79.1%, 380.2% and 30.15%.
Example 3 growth-promoting test of cucumber seedlings by Bacillus amyloliquefaciens, chitosan oligosaccharide and lentinan compounding
The cucumber seeds are subjected to disinfection and seed soaking, then are subjected to seedling raising, and root irrigation treatment by using a chitosan oligosaccharide solution, bacillus amyloliquefaciens XY-13 (CCTCC M2021270) culture solution, a lentinan solution and a compound composition in a period of one leaf and one heart of the cucumber seedlings, and 15ml of liquid is filled into each hole by using a 72-hole plug as an example. Root irrigation is carried out once every 10 days for three times, and seedling growth indexes are counted. The lentinan concentration was set to 400-fold, 700-fold, 1000-fold. Root irrigation with pure water is used as a control, root irrigation treatment is carried out 3 times on the 1 st day, 10 th day and 20 th day of the beginning of treatment, 11 treatments are provided, each treatment is repeated 3 times, 40 plants are treated each, and seedling growth vigor is measured on the 30 th day.
Treatment 1: blank control, water only
Treatment 2: 400-fold lentinan diluent
Treatment 3: 700-fold lentinan diluent
Treatment 4: 1000-fold lentinan diluent
Treatment 5:500mg/L chitosan oligosaccharide solution +10 7 cfu/ml XY-13 culture solution
Treatment 6:500mg/L chitosan oligosaccharide solution +10 7 cfu/ml XY-13+400 times lentinan diluent
Treatment 7:500mg/L chitosan oligosaccharide solution +10 7 cfu/ml XY-13+700 times lentinan diluent
Treatment 8:500mg/L chitosan oligosaccharide solution +10 7 cfu/ml XY-13+1000 times lentinan diluent
TABLE 8 influence of Bacillus amyloliquefaciens, chitosan oligosaccharide and lentinan combinations on cucumber seedling growth vigor
As is clear from Table 8, the treatment with lentinan at different concentrations, 500mg/L chitosan oligosaccharide solution +10 7 cfu/ml XY-13, 500mg/L Chitosan oligosaccharide solution +10 7 cfu/ml XY-13 strain subjected to compounding treatment of lentinan with different concentrationsThe relative growth rate, the stem thickness relative growth rate and the chlorophyll content are all obviously higher than those of the blank control treatment. There was no significant difference in plant height relative growth rate between the other treatments except the placebo treatment. The growth indicators among the lentinan treatments (treatments 2-4) with different concentrations and the growth indicators among the treatments 5 have no obvious difference. Treatment 7 (500 mg/L Chitosan oligosaccharide solution +10 compared to treatment 1 7 cfu/ml XY-13+700 times lentinan diluent), the relative growth rate of the stem and the thick and strong seedling index is highest, and is obviously higher than 61.4 percent and 20.8 percent; treatment 8 (500 mg/L Chitosan oligosaccharide solution +10) 7 cfu/ml XY-13+1000 times lentinan diluent), the plant height relative growth rate and chlorophyll content are highest, the relative treatment 1 is higher than 38.2% and 69.1%, and the effect of treatment 7 is better than that of single lentinan treatment, chitosan oligosaccharide solution and bacillus amyloliquefaciens compound treatment.
TABLE 9 influence of Bacillus amyloliquefaciens, chitosan oligosaccharide and lentinan combinations on root development of cucumber seedlings
As can be seen from Table 9, the root growth index was significantly higher for treatments 5-8 than for treatment 1. The root length, root surface area and root volume of lentinan with different concentrations are all obviously higher than those of the blank control treatment. Wherein treatment 7 (500 mg/L Chitosan oligosaccharide solution +10) 7 cfu/ml XY-13+700 times lentinan dilution) the root length, root surface area, root diameter, root volume effect was best, 47.7%, 140.2%, 32.5%, 171.3% higher than treatment 1.
TABLE 10 influence of Bacillus amyloliquefaciens, chitosan oligosaccharide and lentinan combinations on photosynthesis of cucumber seedlings
As can be seen from Table 10, both the transpiration rate and the net photosynthetic rate of treatment 5 and the treatment (treatments 6 to 8) of lentinan were significantly higher than those of treatment 1. The net photosynthetic rate of lentinan treatment at different concentrations was significantly higher than that of CK treatment. Wherein treatment 7 (500 mg/L Chitosan oligosaccharide solution +10) 7 cfu/ml XY-13+700 times lentinan diluent), the transpiration rate, the net photosynthesis rate and the pore conductance are highest, which are 60.0%, 68.5% and 28.6% higher than those of treatment 1. And the effect of the treatment 7 is better than that of the single lentinan treatment, chitosan oligosaccharide solution and bacillus amyloliquefaciens compound treatment.
TABLE 11 influence of Bacillus amyloliquefaciens, chitosan oligosaccharide and lentinan complexation on matrix chemistry
As can be seen from Table 11, the organic matter, quick-acting nitrogen, quick-acting phosphorus and total nitrogen of lentinan at different concentrations were significantly higher than those of the blank treatment, and the treatment with the 700-fold lentinan diluent was optimal. The matrix chemistry of treatment 5 was significantly higher than treatment 1, except for the fast-acting potassium. The chemical property index of each matrix of the compound treatment of treatment 5 and lentinan is obviously higher than that of treatment 1, wherein treatment 7 (500 mg/L chitosan oligosaccharide solution +10) 7 cfu/ml XY-13+700 times lentinan diluent), the content of the organic matters, quick-acting nitrogen, quick-acting potassium and quick-acting phosphorus treated by the method is highest, and the content of the organic matters, the quick-acting nitrogen, the quick-acting potassium and the quick-acting phosphorus is 32.2 percent, 294.1 percent, 67.9 percent and 38.0 percent higher than that of the organic matters, the quick-acting potassium and the quick-acting phosphorus treated by the method 1. Treatment 6 (500 mg/L Chitosan oligosaccharide solution +10) 7 cfu/ml XY-13+400 times lentinan diluent) treatment had the highest total nitrogen content, 13.2% higher than treatment 1.
As can be seen from tables 1-11, the activity index of plant cells can be enhanced by chitosan oligosaccharide treatment, the growth of roots, stems and leaves can be promoted, the chlorophyll content can be improved, and the photosynthesis of crops can be improved; compared with the combination of single chitosan oligosaccharide, lentinan, chitosan oligosaccharide and bacillus amyloliquefaciens, the compound composition has obvious effect of promoting the growth of cucumber seedlings, is beneficial to the absorption of nutrients and moisture by the cucumber seedlings and the robust growth, and shows obvious synergistic effect. Meanwhile, the compound composition has more remarkable effect of improving matrix soil nutrients.
Example 4 Effect of Chitosan oligosaccharide, bacillus amyloliquefaciens, lentinan Complex composition on Low temperature resistance of cucumber seedlings
In the test, two-leaf Yixindell LD-1 cucumber seedlings are used as test materials, irrigation is uniformly carried out according to each treatment of the growth requirements of crops in the seedling stage, and 1 liter of water is irrigated into one plug (72 holes) at a time. Culturing cucumber seedling in an illumination incubator under normal temperature, irrigating root with clear water as control, and adding 500mg/L chitosan oligosaccharide +10 7 The mixed solution of bacillus amyloliquefaciens and lentinan of which the ratio is 700 is used for preprocessing cucumber seedlings for three days. And (3) carrying out low-temperature stress treatment on the fourth day, and sampling and measuring indicators such as growth vigor, antioxidant enzyme activity and the like from the tenth day. 3 different temperature combinations were set up in the experiment: the day and night temperature of low temperature treatment is controlled to be 20 ℃ and 12 ℃ and 16 ℃ and 8 ℃ by taking the day and night temperature of 28 ℃ and 18 ℃ as a control, and the illumination time and the intensity are uniform. 0.5g of fresh leaves of different treated cucumber seedlings are weighed and MDA content is determined by thiobarbituric acid method. The SOD activity of fresh cabbage leaves is measured by using a Nitrogen Blue Tetrazolium (NBT) method, the POD activity is measured by using a guaiacol method, and the CAT activity is measured by using an ultraviolet spectrophotometry.
Treatment 1: blank control, water only, day and night temperature: 28 ℃/18 DEG C
Treatment 2:500mg/L chitosan oligosaccharide solution +10 7 Bacillus amyloliquefaciens culture solution and 700-fold lentinan diluent
Treatment 3: blank control, water only, day and night temperature: 20 ℃/12 DEG C
Treatment 4:500mg/L chitosan oligosaccharide solution +10 7 Bacillus amyloliquefaciens culture solution and 700-fold lentinan diluent
Treatment 5: blank control, water only, day and night temperature: 16 ℃/8 DEG C
Treatment 6:500mg/L chitosan oligosaccharide solution +10 7 Bacillus amyloliquefaciens culture solution and 700-fold lentinan diluent
As can be seen from fig. 4, the cucumber seedling height significantly decreased with decreasing temperature. Under the same temperature environment, the plant height and the seedling strengthening index of the blank control treatment and the compound composition treatment have no obvious difference, but the stem thickness of the compound composition treatment is obviously higher than that of the blank control treatment by 4.4% at the day and night temperature of 28 ℃/18 ℃, and the chlorophyll content of the compound composition treatment is obviously higher than that of the blank control treatment by 24.7% at the day and night temperature of 20 ℃/12 ℃.
As is clear from FIG. 5, the dry fresh weights of the upper and lower parts of cucumber seedlings tended to decrease with decreasing temperature. Under the same temperature environment, the dry fresh weight of the overground part and the fresh weight of the underground part between the blank control treatment and the compound composition treatment have no obvious difference, and the dry weight of the underground part of the compound composition treatment is obviously higher than that of the blank control treatment by 27.2 percent when the day and night temperature is 20 ℃/12 ℃. Overall, the temperature change has the greatest effect on the fresh weight of the subsurface.
As can be seen from fig. 6, SOD, POD, CAT activity of the formulated compositions was equal to or greater than that of the placebo treatment at each time period, and the MDA content was the opposite. Under the treatment at normal temperature and low temperature, the enzyme activity and MDA content change trend of the compound composition treatment and the blank control treatment at each period are similar, the enzyme activity index of the compound composition treatment is higher than that of the blank control treatment, and the malondialdehyde content is lower than that of the blank control treatment. The POD and SOD activities treated by the compound composition are reduced in 2h under the condition of sub-low temperature treatment, namely at 20 ℃/12 ℃ in day and night, then are greatly increased, peak value is reached in 8h, and the POD and SOD activities treated by the blank control are reduced, slightly increased and then reduced after the low temperature stress treatment. At 8h post treatment, POD, SOD activity of t2 treatment was significantly higher than 38.3%, 77.2% of the blank treatment. The method comprises the steps of carrying out a first treatment on the surface of the CAT activity of both treatment 3 and treatment 4 decreased at 2h, followed by an increase, but treatment 4 increased faster and peaked higher than treatment 3; the MDA content of treatment 4 reached a peak at 2h, then decreased and then slightly increased, but treatment 3 reached a peak at 4h, then decreased, and the MDA accumulation of treatment 4 was significantly lower than that of treatment 3.
Low temperature stress is a major factor limiting the production of greenhouse vegetables. The cucumber is warm in nature and not cold-tolerant, the growth of the cucumber can be obviously inhibited by low-temperature stress, especially the plant height, stem thickness and biomass accumulation of the cucumber are limited, and the chlorophyll content and photosynthesis of the cucumber are influenced. According to the application, the cucumber seedling low-temperature stress treatment test shows that the normal growth of the cucumber seedling is influenced under the low-temperature environment, and the influence is larger as the temperature is lower. The compound composition is used for root irrigation, so that the indexes such as chlorophyll and the dry weight of the underground part of cucumber seedlings can be obviously relieved. Secondly, the low-temperature treatment has influence on the antioxidant enzyme activity and malondialdehyde content in cucumber seedling leaves, and the enzyme activity of the cucumber seedlings is firstly reduced, then increased and then slowly reduced after the cucumber seedlings are subjected to low-temperature stress; the trend of malondialdehyde changes inversely. The enzyme activity of cucumber seedlings treated by adding the compound composition is obviously higher than that of the blank control treatment, and the malondialdehyde content of the cucumber seedlings treated by adding the compound composition is obviously lower than that of the blank control treatment. The root irrigation treatment of the chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan compound composition can improve the low-temperature resistance of cucumber seedlings, and has important significance for cultivating strong cucumber seedlings and ensuring safe overwintering of cucumber seedlings.
EXAMPLE 5 Effect of Chitosan oligosaccharide on Bacillus amyloliquefaciens Activity
Inoculating bacillus amyloliquefaciens single colony into a 50mL triangular flask containing 10mL of LB broth, culturing at 180r/min at 30 ℃ for 24 hours to obtain seed liquid; 1mL of the seed solution was taken in a 250mL triangular flask containing 50mL of LB broth, and 500mg/L of chitosan oligosaccharide solution was added in 1%, 2%, 3% and 4% of the inoculum size (50 mL), and incubated at 180r/min at 30℃for 0, 4, 8, 12, 16, 20, 24, 48 and 72 hours, respectively, to determine OD600.
Treatment 1: treatment with seed Strain alone
Treatment 2: strain +0.5ml 500mg/L chitosan oligosaccharide solution
Treatment 3: strain +1ml 500mg/L Chitosan oligosaccharide solution
Treatment 4: strain +1.5ml 500mg/L Chitosan oligosaccharide solution
Treatment 5: strain +2ml 500mg/L chitosan oligosaccharide solution
Referring to FIG. 7, the results of activity of chitosan oligosaccharide addition on Bacillus amyloliquefaciens show that chitosan oligosaccharide has an accelerating effect on bacterial proliferation with the increase of time from 8 hours later.
Claims (2)
1. The application of a chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan compound composition in promoting cucumber seedling growth is characterized in that: the compound composition is in a liquid form, wherein the concentration of the chitosan oligosaccharide is 100-700mg/L, and the viable count of the bacillus amyloliquefaciens is 10 5 -10 8 cfu/mL, wherein the lentinan is 400-1000 times of lentinan diluent.
2. The use of the compound composition of chitosan oligosaccharide, bacillus amyloliquefaciens and lentinan in promoting cucumber seedling growth, which is characterized in that: the concentration of the chitosan oligosaccharide is 500mg/L, and the viable count of the bacillus amyloliquefaciens is 10 7 cfu/mL, the lentinan is 700 times lentinan diluent.
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| CN115261282A (en) * | 2022-08-26 | 2022-11-01 | 甘肃省科学院生物研究所 | Bacillus amyloliquefaciens and application thereof in cucumber disease prevention and control |
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| CN103190404A (en) * | 2013-04-02 | 2013-07-10 | 中国农业科学院植物保护研究所 | Composition for preventing and treating plant nematode and virus diseases, and application thereof |
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