Application of sporibospora lyrata ZRQ01 in plant growth promotion

Abstract

The invention belongs to the technical field of microbial fermentation, and particularly relates to application of sporidiobolus sp. ZRQ01 in plant growth promotion. The sporidiobolus sp. ZRQ01 has been preserved in the Guangdong Microbial Culture Collection Center on May 22, 2025, the address is the 5th floor of the building 59, No.100 Courtyard, Xianlie Middle Road, Guangzhou, and the preservation number is CDMCC No: 66380. The invention provides the application of the sporidiobolus sp. ZRQ01 in plant growth promotion, and a growth-promoting microbial inoculum is developed by taking low-value agricultural products such as cabbages as raw materials, and has the characteristic of low cost; the data result of the embodiment of the invention shows that the growth-promoting microbial inoculum prepared by the invention can greatly improve the growth of soybeans.

Description

Technical Field

[0001] This application belongs to the field of microbial fermentation technology, specifically relating to the application of ZRQ01 yeast in plant growth promotion. Background Technology

[0002] Globally, approximately 1.3 billion tons of agricultural waste are generated annually. Leafy vegetable waste (such as cabbage and broccoli) is particularly problematic due to its high water content, easy decomposition, and high transportation costs. Traditional treatment methods primarily involve landfilling or composting, which not only wastes resources but also contributes to greenhouse gas emissions. Although some technologies attempt to convert waste into organic fertilizer, their complex composition, unstable fertilizer effectiveness, and the risk of heavy metal accumulation hinder large-scale application. Therefore, developing high-value utilization technologies for low-value agricultural waste has become a core requirement for sustainable agricultural development.

[0003] Currently, the nitrogen-fixing properties of legumes (such as soybeans) make them important economic crops, but in actual production, they still face problems such as long growth cycles, weak stress resistance, and yield bottlenecks. At present, plant growth promoters mainly include two categories: (1) chemically synthesized ones, which have problems such as high cost, soil residue, and ecotoxicity. (2) microbial fermentation ones, which are environmentally friendly, but the functions of the strains are limited and the composition is unclear.

[0004] Therefore, there is an urgent need to develop a plant growth promoter that uses agricultural waste as raw material, undergoes targeted fermentation by specific functional strains, and has clearly defined components and effects, in order to overcome the triangular contradiction between resources, environment, and agricultural production. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide an application of ZrQ01 yeast in plant growth promotion, specifically adopting the following technical solution: In a first aspect, the present invention provides *Saccharomyces cerevisiae* (…). Sporidiobolus pararoseus The application of ZRQ01 in plant growth promotion: ZRQ01 was deposited on May 22, 2025 at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number CDMCC No: 66380.

[0006] The ITS sequence of the *Zygosaccharomyces cerevisiae* ZRQ01 is shown in SEQ ID NO: 1.

[0007] SEQ ID NO: 1: .

[0008] As a further preferred embodiment, the plant includes a legume.

[0009] As a further preferred embodiment, the legume includes soybean.

[0010] Secondly, the present invention provides a growth-promoting microbial agent for crops, the growth-promoting microbial agent comprising the above-mentioned *Saccharomyces cerevisiae* ZRQ01 or its fermentation broth.

[0011] As a further preferred embodiment, the growth-promoting agent is obtained by fermentation of a culture medium containing cabbage.

[0012] As a further preferred embodiment, the mass ratio of cabbage to water in the culture medium is 5%-20%:1.

[0013] As a further preferred embodiment, the culture medium also includes Mn 2+ element.

[0014] As a further preferred embodiment, the content of the *Zygosaccharomyces cerevisiae* ZRQ01 is 4%.

[0015] Thirdly, a method for preparing a growth-promoting microbial agent for crops, characterized by comprising the following steps: Add cabbage to a container containing Mn 2+ In the culture medium containing the elements, sterilize at 121°C for 15 minutes, then add Cyclospora yeast and culture to obtain the growth-promoting bacterial agent for crops.

[0016] As a further preferred embodiment, the cabbage added is 5%-20% of the water.

[0017] The beneficial effects of this invention are as follows: This invention provides the application of *Saccharomyces cerevisiae* ZRQ01 in plant growth promotion, and develops a growth-promoting agent using low-value agricultural products such as cabbage as raw materials, which has the characteristics of low cost; according to the data results of the embodiments of this invention, the growth-promoting agent prepared by this invention can greatly improve the growth of soybeans. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 The image shown is a colony morphology diagram of *Saccharomyces cerevisiae* ZRQ01. Figure 2 The image shows the fermentation broth of *Syngonium spp.* ZRQ01 inoculated into a medium containing 5%-20% cabbage and cultured for 8 days. Figure 3 The figure shows the growth characteristics of soybean seedlings cultured in a culture medium containing 5%-20% fermentation liquid of ZRQ01 for 30 days. Figure 4 The image shows the phylogenetic tree of the ZRQ01 strain of *Saccharomyces cerevisiae*. Detailed Implementation

[0020] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0021] Example 1 Cyclosporum tobira ( Sporidiobolus pararoseus Screening and identification of ZRQ01 (1) Isolation and culture of strains 1.0 g of mealworm feces that had ingested PLA powder was added to 50 mL of LB liquid medium (PLA inorganic salt medium) containing 2.0 g of 300 mesh PLA powder. The medium was incubated at 30 ℃ and 220 r / min with shaking for 48 h. Then, 1.0 mL of the enriched solution was serially diluted to 10⁻⁶. -3 10 -4 10 -5 Different dilutions were used, with 50 µL of each dilution spread onto solid culture medium containing PLA 350 mesh powder and incubated at 37 °C for 3 days for initial screening. The selected bacteria were then inoculated onto cellulose mononutrient culture plates and incubated for 3 days. The growth of the bacteria is shown below. Figure 1 As shown, its colonies are red.

[0022] The colonies growing well on the plates were numbered and isolated and purified multiple times. The purified strains were then cultured in a 37°C incubator for further isolation and purification. The selected microorganisms were then preserved and sequenced.

[0023] The selected ZRQ01 yeast strain was observed under a microscope to be round or oval, exhibiting budding reproduction, with a smooth surface and visible shiny vesicles.

[0024] (2) Molecular identification The ITS sequence of the screening bacteria was amplified, and the target gene sequence was obtained. Nucleotide homology was compared with the sequence in GenBank using BLAST.

[0025] The sequence results were compared with the NCBI database, and it was found that the similarity of strain ZRQ-08 was over 99%. Sporidiobolus pararoseus From the phylogenetic tree ( Figure 4 From the above, it can be concluded that strain ZRQ-08 is similar to strain JQ425362.1. Sporidiobolus pararoseus Within the same evolutionary branch, the self-spreading support rate is 99%, and the sequence homology between the two is over 99%. Based on morphological characteristics, it can be inferred that the screened strain ZRQ-08 is... Sporidiobolus pararoseus One of the sequences. The sequence has been registered in GenBank (nucleic acid sequence data: SUB1735606155473), and in this study it is named Sporidioboluspararoseu ZRQ01.

[0026] Based on the physiological and morphological characteristics of the fungus, its species was determined, and it was ultimately identified and named *Saccharomyces cerevisiae*. Sporidiobolus pararoseusZRQ01, the strain was deposited at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No: 66380.

[0027] Table 1. Physiological and biochemical characteristics Example 2 Cyclosporum tobira ( Sporidiobolus pararoseus Determination of the growth capacity of soybeans using ZRQ01 (1) Preparation of fermentation broth of Saccharomyces cerevisiae ZRQ01, the specific process is as follows: Wash and chop the cabbage, and add Mn at amounts of 5%, 10%, 15% and 20% respectively. 2+ Distilled water containing 0.025 mg / 100 mL of elemental compounds (where 5%-20% refers to the cabbage's weight in the distilled water being 5%-20%) was sterilized at 121℃ for 15 minutes. Then, 4% of *Saccharomyces cerevisiae* culture, which had been incubated for 48 hours, was added. The mixture was then incubated at 25℃ with shaking at 180 rpm for 8 days to obtain the fermentation broth. The results are as follows: Figure 2 As shown. After fermentation, the fermentation broths cultured with 5%-20% cabbage content all changed color, indicating that *Zygosporum tobira* ZRQ01 grew well. Specifically, the fermentation broths with 5%-20% cabbage content were pink, light yellow, orange-yellow, and orange-red, respectively. This invention also investigated the fermentation effect with 25% cabbage addition; the color of the fermentation broth with 25% cabbage addition was significantly lighter than that with 20% cabbage addition, indicating that under the culture conditions of 20% cabbage, the *Zygosporum tobira* inoculum amount was 4%, M... 2+ Shaking culture under certain conditions for 8 days is beneficial for the accumulation of the metabolite β-carotene. Subsequently, 20% cabbage was added for fermentation.

[0028] (2) The fermentation broth of ZRQ01 yeast prepared above was used to culture soybean plants. To investigate the effect of the above fermentation broth on soybean growth promotion, the *Saccharomyces cerevisiae* ZRQ01 fermentation broth prepared with 20% cabbage content was filtered and diluted to different concentrations. The effects of the fermentation broth on soybean growth factors were observed. The broth was diluted with purified water. Group 1: 5% (v / v) fermentation broth; Group 2: 10% (v / v) fermentation broth; Group 3: 15% (v / v) fermentation broth; Group 4: 20% (v / v) fermentation broth; Group CK: no fermentation broth added, only purified water added. The liquid addition depth was kept consistent, and liquid was added midway to ensure that the soybean roots were fully submerged in the liquid. After 30 days of cultivation, the growth indicators of the soybean seedlings are shown in Table 2. Figure 3 As shown; Table 2 The data in the table above shows that adding 5%-20% fermentation broth to the soybean culture medium increases the plant height, stem diameter, chlorophyll content, and number of fruits in soybean seedlings. Furthermore, the malondialdehyde (MDA) and catalase (CTA) content in soybean seedlings is significantly increased, indicating that the fermentation broth formed by fermenting cabbage with Saccharomyces cerevisiae ZRQ01 has a certain promoting effect on the growth of soybean plants.

[0029] The embodiments of this application have been described above with reference to the accompanying drawings. Specific examples have been used to illustrate the principles and implementation methods of this application. The description of the above embodiments is only for the purpose of helping to understand the core ideas of this application. However, this application is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this application without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of this application.

Claims

1. *Saccharomyces cerevisiae* ( Sporidiobolus pararoseus The application of ZRQ01 in promoting plant growth is characterized by, The *Saccharomyces cerevisiae* ZRQ01 was deposited on May 22, 2025, at the Guangdong Provincial Center for Microbial Culture Collection, located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number CDMCC No: 66380.

2. The application according to claim 1, characterized in that, The plants mentioned include legumes.

3. The application according to claim 2, characterized in that, The legumes mentioned include soybeans.

4. A growth-promoting bacterial agent for crops, characterized in that, The growth-promoting agent includes the *Saccharomyces cerevisiae* ZRQ01 or its fermentation broth as described in claim 1.

5. The growth-promoting bacterial agent according to claim 4, characterized in that, The growth-promoting agent is obtained by fermentation of a culture medium containing cabbage.

6. The growth-promoting bacterial agent according to claim 5, characterized in that, The mass ratio of cabbage to water in the culture medium is 5%-20%:

1.

7. The growth-promoting bacterial agent according to claim 6, characterized in that, The culture medium also includes Mn. 2+ element.

8. The growth-promoting bacterial agent according to claim 7, characterized in that, The content of the *Zygosaccharomyces cerevisiae* ZRQ01 is 4%.

9. A method for preparing a growth-promoting bacterial agent for crops, characterized in that, Includes the following steps: Add cabbage to a container containing Mn 2+ In the culture medium containing the elements, sterilize at 121°C for 15 minutes, then add Cyclospora yeast and culture to obtain the growth-promoting bacterial agent for crops.

10. The preparation method according to claim 9, characterized in that, The amount of cabbage added is 5%-20% of the water content.

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