CN111512824A - Method for promoting growth of greening plants by using composite microbial inoculum - Google Patents
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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
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Abstract
The invention discloses a method for promoting the growth of greening plants by using a complex microbial inoculum, which comprises the steps of inoculating endophytic fungi and arbuscular mycorrhizal fungi on the roots of the greening plants in a mixed manner, and then planting the inoculated greening plants. The invention has good promotion effect on the survival and growth of green plants, and can be popularized and applied in ecological restoration projects in adverse environments such as open coal mines and the like.
Description
Technical Field
The invention belongs to the field of ecological restoration, and relates to a method for promoting growth of green plants by using a complex microbial inoculum.
Background
China is rich in coal resources and is the first coal producing country and consuming country in the world. The mining process of the open pit coal mine causes large-area land resource damage, causes the problems of soil basic structure damage, nutrient substance scarcity, soil microorganism reduction and the like, further causes slow growth of planted vegetation and low resistance stability, and leads to increasingly sharp reduction of local vegetation and biological diversity. The restoration succession ability of the natural restoration of the mining area alone is not enough to restore, and the soil treatment and vegetation construction of the mining area are one of the important contents for the ecological improvement of the mining area.
The technologies applied in the ecological restoration process can be divided into two major types, namely biotechnology and non-biotechnology. Wherein, the non-biological repairing means has fussy operation and high economic cost, and the single use can not thoroughly solve the problem of the soil environment deterioration such as mining areas and the like; the traditional microbial remediation (i.e. single strain inoculation) technology has the disadvantages of generally insignificant effect and small effect on the construction of microbial community structure and function.
At present, the plant-microorganism combined repair technology is an effective repair technology. Namely, on the basis of single plant or microbial remediation, the plants and the microbial inoculum are co-cultured to form a joint symbiotic system, so that respective advantages are fully exerted, and the defect of single plant or microbial remediation is effectively improved. In a plant-microorganism combined repair technology system, microorganisms play an important role in aspects of nutrition absorption, environmental adaptation, disease resistance and the like of plants, and in turn, the plants can provide carbon sources and energy substances required by growth and metabolism for the associated microorganisms, so that the whole ecological system is regulated and controlled. The technology does not produce secondary pollution, and has low cost and high efficiency.
Arbuscular Mycorrhizal Fungi (AMF) are soil microorganisms which can penetrate epidermal cells of host plant roots to form infection structures such as arbuscular, vesicular, intraradicular hyphae and spores and provide mineral nutrient elements for plants through mycorrhizal symbiota and simultaneously exchange carbon sources for growth of the plants. The AMF has strong ecological adaptability and can form symbiont with more than 80% of plants and about 90% of vascular plant root systems on land. In recent years, more and more research results show that AMF can significantly improve the growth and drought resistance of host plants, promote the absorption of the plants to mineral substances, and improve the water metabolism and soil physicochemical properties of the plants. The application of AMF has obvious promotion effect on ecological restoration and reconstruction in severe environment. Endophytes (endophytes) are those microorganisms that live inside various tissues and organs of healthy plants at some or all of their life history without causing significant plant disease, including endophytes, and endophytes. The plant endophyte lives in a special environment in a plant body for a long time and is co-evolved with the plant to establish a joint relationship. On the one hand, plants provide the endophytes with the nutrients and energy necessary for growth; on the other hand, endophytes can influence plants by their own metabolic products or by means of signal transduction. The endophytic fungi can produce various secondary metabolites with biological activity, thereby directly or indirectly influencing the growth and development, propagation, environmental adaptability and the like of host plants. Endophytes are of great interest as a new microbial resource.
Disclosure of Invention
The invention aims to solve the technical problems of soil structure damage, nutrient deficiency, difficult vegetation survival and the like in ecological greening treatment.
In order to solve the technical problems, the invention provides a method for promoting the growth of green plants by using a compound microbial inoculum based on a plant-microorganism combined restoration concept, which comprises the steps of inoculating endophytic fungi and arbuscular mycorrhizal fungi on the roots of the green plants in a mixed manner, and then planting the inoculated green plants.
In some embodiments, the above method, the mixed inoculation is performed by soaking the root system of the green plant in 1 × 105-1×10710-15 hours in the endophytic fungi bacterial liquid with cfu/mL concentration, and then wrapping 15-25g of arbuscular mycorrhizal fungi bacterial agent on the root, for example, soaking the root of the green plant in 1 × 106The bacterial liquid of endophytic fungi with the concentration of cfu/mL is 12 hours, and the root of the bacterial liquid is wrapped with 20g of arbuscular mycorrhizal fungi microbial inoculum, and the bacterial liquid can be prepared by using distilled water, phosphate buffer solution or normal saline.
In some embodiments, in any of the methods described above, the endophytic fungus is zygospora circinatum.
In some embodiments of any of the methods described above, the arbuscular mycorrhizal fungus is one or more of the group consisting of mycosphaerella pusilla (Funneliformis mossse), rhizospora radicicola (Rhizophagus intraradicalis), acanthospora gracilis (Acaulospora scrobicula), and variospora ivorum ivory (diverisia eburana), preferably variospora ivorum (diverisia eburana).
In some embodiments, the planting method is a conventional green seedling planting method, and necessary early-stage management and care such as watering and pest control are performed.
In some embodiments, the method of any one of the above, wherein the green plant is Stipagrandis arundinacea (Stipagrandis p.
In order to solve the technical problems, the invention also provides a microbial inoculum which comprises endophytic fungi and arbuscular mycorrhizal fungi.
In some embodiments, in the above microbial agent, the endophytic fungus and arbuscular mycorrhizal fungus are packaged separately.
In some embodiments, the inoculant of any one of the above, wherein the endophytic fungus is Scytalidium circinatum.
In some embodiments, the inoculant described in any one of the above paragraphs, wherein the arbuscular mycorrhizal fungus is one or more of the group consisting of mycosphaerella pusilla (Funneliformis mossse), rhizospora radicicola (Rhizophagus intraradicalis), acauliospora dricola (acaulisporidia scrobicula), and variospora ivorangii (diverisispora eburana), preferably variospora ivorangii (diverisispora eburana).
In order to solve the technical problems, the invention also provides application of the endophytic fungi and arbuscular mycorrhizal fungi in promoting growth and/or ecological restoration of greening plants.
In some embodiments, in the above use, the endophytic fungus is Scytalidium circinatum.
In some embodiments, in the use of any of the above, the arbuscular mycorrhizal fungus is one or more of the group consisting of mycosphaerella pusilla (Funneliformis mossse), rhizospora radicicola (Rhizophagus intraradicles), acanthospora gracilis (Acaulospora scrobicula), variospora ivorum (diverisia eburana), preferably variospora ivorum (diverisia eburana).
In some embodiments, in the use of any of the above, the green plant is Stipagrandis p.
The principle of the invention is that the survival rate of the greening plants is improved, the underground and overground biomass and the plant height are increased by utilizing the synergistic symbiotic action of the composite microbial inoculum and the greening plants, and the ecological greening effect is further obviously improved.
The invention has the advantages that:
1. the invention establishes a vegetation-compound microorganism mutualism symbiosis restoration system by utilizing the compound microorganism bacterium agent, so that the vegetation and the microorganism respectively exert the advantages and functions, the survival rate of transplanted seedlings is improved, and the invention has a potential good application prospect in the fields of open coal mine ecological restoration and the like.
2. The compound microbial agent adopted by the invention is lifelong, can continuously provide inorganic nutrition such as N, P and the like for plants, promotes the growth of transplanted seedlings, and is added once to benefit lifelong.
3. The added compound microbial inoculum can increase the amount of soil microorganisms, improve the soil environment and improve the soil fertility, and can create good conditions for the growth of other surrounding greening plants and natural falling plants.
4. The method has the advantages of simple operation, low comprehensive cost, no pollution in the process and good popularization and application values.
Drawings
FIG. 1 shows the above-ground biomass measurement results of Achillea alpina of each treatment group.
FIG. 2 shows the underground biomass measurement results of the individual treatment groups of Achillea alpina.
FIG. 3 shows the results of measuring the plant height of Achillea alpina in each treatment group.
FIG. 4 shows the mycorrhizal infection rate of the stipa lanceolata in each treatment group.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The present invention will be further described with reference to the following examples. It should be understood that the following examples are illustrative only and are not intended to limit the scope of the present invention.
Section of convoluted sporulata (Scytalidium circinatum) is disclosed in the literature "variety study of species of hyphomycetes in soils in the plateau area of gunns. *** [ D ]. Shandong university of agriculture, 2008", publicly available from the university of inner Mongolia.
Microcystis ivorangii (Diclaspora eburnea) is purchased from a germplasm resource library BGC of arbuscular mycorrhizal fungi, which is called BGC-XJ06B for short.
Rhizopus intraradicans (Rhizophagus intraradics) is purchased from a germplasm resource library BGC of arbuscular mycorrhizal fungi, which is called BGC-BJ09 for short.
Tunica morsella (Fusneliformis mossea) is purchased from a germplasm resource library BGC of arbuscular mycorrhizal fungi, which is called BGC-BJ01 for short.
The aureobasidium tenue (Acaulospora scrobicula) is purchased from a germplasm resource library BGC of arbuscular mycorrhizal fungi, which is called BGC-HK02A for short.
The Glomus minor (Glomus etunicatum) is purchased from a germplasm resource library BGC of arbuscular mycorrhizal fungi, which is called BGC-XJ03C for short.
Examples
(1) Process layout
Transplanting consistent, strong and uniform seedlings of Stipa macrocarpa (Stipa grandis P. Smirn.) into a flowerpot filled with 3kg of sterilized mixed soil, wherein the mixed soil is prepared by mixing vermiculite and nutrient soil according to a volume ratio of 1:1, the specification of the flowerpot is 15cm multiplied by l0cm multiplied by 13cm, soaking the flowerpot in 0.5% potassium permanganate solution overnight, and then washing the flowerpot clean by tap water.
A total of 10 treatment groups were set, including Control (CK), Scytalidium circinatum (E +), Scytalidium circinatum) + Fagaku (Fusneliformis mosseae) (E + BGC-BJ01), Scytalidium circinatum) + Rhizophora rhizogenes (Rhizophylloides) (E + BGC-BJ09), Scytalidium circinatum) + Rhizophyllum rhizogenes (Rhizophylloides) (E + BGC-BJ09), Scytalidium circinatum) + Acaureobasidium fascicularis (Acaulospora bicula) (E + BGC-HK A), Scytalidium circinatum cicola (BGC-5932), Rhizophyllum forbespora (Schizophyllum), Rhizophyllum forbescus (Schizophyllum minor sporotrichuria) (E + BGC-5932), and Rhizophyllum fornica (BGC-5932), and Rhizophyllum fornica (Rhizophyllum minor rhizopus) (BGC-5932). Each treatment group was replicated 10 times, and 3 seedlings were planted per pot.
The specific treatments performed by each treatment group were as follows:
CK: firstly, 2.5kg of sterilized mixed soil is put into a flowerpot, and the stipa capillata seedlings are planted into the flowerpot; then 0.5kg of sterilized mixed soil is paved to cover the roots of the stipa longipes seedlings; compacting the covering soil and watering thoroughly.
E +. soaking the root of the stipa glauca seedling in Scytalidium circinatum (Scytalidium circinatum) bacterial liquid (concentration is 1 × 10)6cfu/mL) for 12 h; 2.5kg of sterilized mixed soil is put into a flowerpot, and then the seedlings soaked with the bacterial liquid are put into the flowerpot; then 0.5kg of sterilized mixed soil is paved, the covering soil is compacted and the mixture is watered thoroughly.
E + BGC-BJ01 soaking root of Dactylis glomerata seedling in Scytalidium circinatum (Scytalidium circinatum) bacterial solution (concentration 1 × 10)6cfu/mL) for 12 h; 2.5kg of sterilized mixed soil is put into a flowerpot, the seedling soaked with the bacterial liquid is put into the flowerpot, and 20g of AM bacterial agent (Moxidou cystoma bacterial agent BGC-BJ01) is applied to the periphery of the root of the transplanted stipa capillata seedling; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
E + BGC-BJ09 soaking root of Dactylis glomerata seedling in Scytalidium circinatum (Scytalidium circinatum) bacterial solution (concentration 1 × 10)6cfu/mL) for 12 h; 2.5kg of sterilized mixed soil is put into a flowerpot, the seedling soaked with the bacterial liquid is put into the flowerpot, and 20g of AM bacterial agent (root cyst mould agent BGC-BJ09 in the root) is applied to the periphery of the root of the transplanted stipa capillata seedling; then 0.5kg of sterilized mixed soil is pavedCovering the AM microbial inoculum and the roots of the stipa capillata seedlings with soil; compacting the covering soil and watering thoroughly.
E + BGC-HK02A soaking root of Dactylis glomerata seedling in Scytalidium circinatum (Scytalidium circinatum) bacterial solution (concentration 1 × 10)6cfu/mL) for 12 h; 2.5kg of sterilized mixed soil is put into a flowerpot, the seedling soaked with the bacterial liquid is put into the flowerpot, and 20g of AM microbial inoculum (a fine concave non-stem cyst mold agent BGC-HK02A) is applied to the periphery of the root of the transplanted stipa capillata seedling; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
E + BGC-XJ06B soaking root of Dactylis glomerata seedling in Scytalidium circinatum (Scytalidium circinatum) bacterial solution (concentration 1 × 10)6cfu/mL) for 12 h; 2.5kg of sterilized soil is put into a flowerpot, the seedling soaked with the bacterial liquid is put into the flowerpot, and 20g of AM microbial inoculum (heterospora ivory BGC-XJ06B) is applied to the periphery of the root of the transplanted seedling of the stipa arundinacea; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
BGC-BJ 09: firstly, 2.5kg of sterilized mixed soil is put into a flowerpot; applying 20g of AM microbial inoculum (BGC-BJ09) around the roots of the transplanted stipa capillata seedlings; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
BGC-BJ 01: firstly, 2.5kg of sterilized mixed soil is put into a flowerpot; applying 20g of AM microbial inoculum (BGC-BJ01) around the roots of the transplanted stipa capillata seedlings; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
BGC-HK 02A: firstly, 2.5kg of sterilized mixed soil is put into a flowerpot; applying 20g of AM microbial inoculum (BGC-HK02A) around the roots of the transplanted stipa virgata seedlings; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
BGC-XJ 03C: firstly, 2.5kg of sterilized mixed soil is put into a flowerpot; applying 20g of AM microbial inoculum (BGC-XJ03C) around the roots of the transplanted stipa capillata seedlings; then 0.5kg of sterilized mixed soil is paved to cover the AM microbial inoculum and the roots of the stipa capillata seedlings; compacting the covering soil and watering thoroughly.
(2) Maintenance management
Each treatment group was watered once every 4 days with 100mL each time, so that the water content was controlled to 60% of the maximum water holding capacity of the soil, the photoperiod was 14L:10D, the culture temperature was 27 ℃ and the air humidity was controlled to 60%.
(3) Sampling assay
After three months, the survival rate, the aboveground biomass, the underground biomass and the plant height of the Dactylis glomerata in the experimental group E +, E + BGC-BJ01, E + BGC-BJ09, E + BGC-HK02A, E + BGC-XJ06B, BGC-BJ09, BGC-BJ01, BGC-HK02A, BGC-XJ03C and the control group (CK) are measured, and the results are respectively shown in Table 1, figure 2 and figure 3. Meanwhile, 3 pots of plants are respectively selected from the experimental group E +, E + BGC-BJ01, E + BGC-BJ09, E + BGC-HK02A, E + BGC-XJ06B, BGC-BJ09, BGC-BJ01, BGC-HK02A, BGC-XJ03C and the control group (CK), and the root system of the stipa glauca is harvested and washed clean by deionized water. Randomly selecting 5 root segments with the length of lcm for each plant, judging infection grade number by a visual method, and according to the percentage of the minor root branching number in the total minor root number, totally 5 grades and 0 grade: 0-10%, grade 1: 10% -19%, grade 2: 20% -29%, grade 3: 30% -39%, grade 4: 40% -49%, grade 5: more than 50 percent. And (3) measuring the mycorrhiza infection rate by adopting a counting statistical method: the infection rate of mycorrhiza is (number of mycorrhiza infection sections/number of total root sections) × 100%.
(4) Results
Table 1 shows that compared with the survival rate of 86.67 percent of a control group, the survival rate of the stipa baicalensis is improved by the experimental group inoculated with the microbial inoculum (the survival rate is over 90 percent), and the survival rate reaches 100 percent after three treatments of E + BGC-BJ01, E + BGC-HK02A and E + BGC-XJ06B under the combined use condition of the two microbial inoculants.
TABLE 1 survival rate of plants table
FIG. 1 shows that the above-ground biomass of the stipa glomerata is increased by the experimental groups inoculated with the microbial inoculum, and the above-ground biomass of the experimental groups E +, E + BGC-BJ01, E + BGC-BJ09, E + BGC-HK02A, E + BGC-XJ06B, BGC-BJ09, BGC-BJ01, BGC-HK02A and BGC-XJ03C is increased by 11%, 12%, 8%, 4%, 31%, 6%, 4%, 11% and 2% respectively relative to the control group (CK), wherein the above-ground biomass of the experimental group E + BGC-XJ06B is increased by 31% to the maximum.
FIG. 2 shows that the underground biomass of the Dactylis glomerata is increased by the experimental groups inoculated with the microbial inoculum, and the underground biomass of the Dactylis glomerata is increased by 31%, 42%, 34%, 6%, 43%, 34%, 6%, 12% and 31% respectively by the experimental groups E +, E + BGC-BJ01, E + BGC-BJ09, E + BGC-HK02A, E + BGC-XJ06B, BGC-BJ09, BGC-BJ01, BGC-HK02A and BGC-XJ03C compared with the control group (CK), wherein the underground biomass of the experimental group E + BGC-XJ06B is increased by 43% and the underground biomass of the experimental group E + BGC + BJ01 is increased by 42% secondly.
FIG. 3 shows that the height of the plants of the stipa longissima is increased in all the experimental groups inoculated with the microbial inoculum, and the heights of the plants of the stipa longissima of the experimental groups E +, E + BGC-BJ01, E + BGC-BJ09, E + BGC-HK02A, E + BGC-XJ06B, BGC-BJ09, BGC-BJ01, BGC-HK02A and BGC-XJ03C are respectively increased by 28%, 33%, 26%, 19%, 35%, 4%, 34% and 27% compared with the control group (CK), wherein the heights of the plants of the stipa longissima of the experimental group E + BGC-XJ06B are increased by 35% most.
FIG. 4 shows that the mycorrhizal infection rate of the stipa glomerata is improved in all the experimental groups inoculated with the microbial inoculum, wherein the experimental groups E +, E + BGC-BJ01, E + BGC-BJ09, E + BGC-HK02A, E + BGC-XJ06B, BGC-BJ09, BGC-BJ01, BGC-HK02A and BGC-XJ03C are respectively improved by 9%, 26%, 30%, 7%, 35%, 5%, 7%, 29% and 17% compared with the control group (CK). The infection rate of the experimental group E + BGC-XJ06B is the highest, and is 35%, the infection rate belongs to 3-grade infection, and except for E + BGC-BJ09, the infection rate of other experimental groups is lower than 30%.
In conclusion, the inoculation inoculants all improve the survival rate of the stipa macrophylla plants, the aboveground biomass, the underground biomass and the plant height, and increase the infection rate of the stipa macrophylla, wherein each index of the experimental group of the Scytalidium circinatum, the variegated ascospora curcas (Diversispora eburnea) (E + BGC-XJ06B) is obviously greater than that of the control group (CK) and is higher than that of other experimental groups, and the inoculants can be selected as the optimal strain combination after comprehensive evaluation.
Claims (10)
1. A method for promoting the growth of greening plants by using a compound microbial inoculum comprises the steps of inoculating endophytic fungi and arbuscular mycorrhizal fungi on the roots of the greening plants in a mixed manner, and then planting the inoculated greening plants.
2. The method as set forth in claim 1, wherein the mixed inoculation is carried out by immersing root systems of the greening plants in 1 × 105-1×10710-15 hours in the endophytic fungi bacterial liquid with cfu/mL concentration, and then 15-25g of arbuscular mycorrhizal fungi microbial inoculum is wrapped at the root of the endophytic fungi bacterial liquid.
3. The method according to claim 1 or 2, characterized in that: the endophytic fungus is Scytalidium circinatum.
4. A method according to any one of claims 1-3, characterized in that: the arbuscular mycorrhizal fungi is one or more of the group consisting of moniliniella bursal (fusanformis mossseae), rhizospora intraradicicola (Rhizophagus intraradics), Acaulospora scrophularis (Acaulospora scrobicula), and variospora ivora ivoides (dichrsispora eburinea), preferably variospora ivora ivoloris (dichrsispora eburinea).
5. A microbial preparation comprises endophytic fungi and arbuscular mycorrhizal fungi.
6. The microbial inoculum of claim 5, wherein: the endophytic fungi and arbuscular mycorrhizal fungi are packaged independently.
7. The microbial inoculum according to claim 5 or 6, characterized in that: the endophytic fungus is Scytalidium circinatum.
8. The microbial inoculum according to any one of claims 5 to 7, wherein: the arbuscular mycorrhizal fungi is one or more of the group consisting of moniliniella bursal (fusanformis mossseae), rhizospora intraradicicola (Rhizophagus intraradics), Acaulospora scrophularis (Acaulospora scrobicula), and variospora ivora ivoides (dichrsispora eburinea), preferably variospora ivora ivoloris (dichrsispora eburinea).
9. Application of endophytic fungi and arbuscular mycorrhizal fungi in promoting growth and/or ecological restoration of green plants.
10. Use according to claim 9, characterized in that: the endophytic fungus is Scytalidium circinatum; and/or
The arbuscular mycorrhizal fungi is one or more of the group consisting of moniliniella bursal (fusanformis mossseae), rhizospora intraradicicola (Rhizophagus intraradics), Acaulospora scrophularis (Acaulospora scrobicula), and variospora ivora ivoides (dichrsispora eburinea), preferably variospora ivora ivoloris (dichrsispora eburinea).
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| CN115486309A (en) * | 2022-09-02 | 2022-12-20 | 中国农业科学院蔬菜花卉研究所 | Method for promoting rose growth by using arbuscular mycorrhizal fungi |
| CN116171808A (en) * | 2022-11-10 | 2023-05-30 | 云南大学 | Method for repairing degraded grassland by combining arbuscular mycorrhizal fungi and bacteria |
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| CN113950923A (en) * | 2021-10-18 | 2022-01-21 | 岭南生态文旅股份有限公司 | Method for improving continuous cropping resistance of peach trees |
| CN115486309A (en) * | 2022-09-02 | 2022-12-20 | 中国农业科学院蔬菜花卉研究所 | Method for promoting rose growth by using arbuscular mycorrhizal fungi |
| CN116171808A (en) * | 2022-11-10 | 2023-05-30 | 云南大学 | Method for repairing degraded grassland by combining arbuscular mycorrhizal fungi and bacteria |
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