Application of streptomyces aloglaucus in sugarcane preservation

Abstract

This invention provides the application of *Streptomyces alogie* in sugarcane preservation, relating to the field of sugarcane preservation technology. Sugarcane tops and leaves treated with *Streptomyces alogie* fermentation broth can effectively slow down the loss of chlorophyll and water, exhibiting a good preservation effect. This further slows down the loss of moisture and nutrients when sugarcane tops and leaves are used as silage, extending the storage period to 10 days, while remaining safe for humans and animals. Sugarcane stalks treated with *Streptomyces alogie* fermentation broth can slow down the rate of decay and spoilage during transportation and storage, extending the storage period to 30 days, thus achieving the goal of long-term sugarcane preservation and providing significant economic and social benefits for industrial development.

Description

Technical Field

[0001] This invention relates to the field of sugarcane preservation technology, specifically to the application of Streptomyces alogie in sugarcane preservation. Background Technology

[0002] Sugarcane is the primary raw material for sugar production, and its byproducts can be used to produce ethanol, feed, fertilizer, and fuel, making it an important raw material and strategic resource for light industry, chemical industry, and energy. Furthermore, sugarcane tops and leaves are rich in amino acids, minerals, vitamins, organic acids, and a certain amount of sucrose and reducing sugars, among other nutrients. Its crude protein content is also higher than that of corn, wheat, rice, and soybean straw, making it a high-quality roughage and silage resource for ruminants such as cattle and sheep.

[0003] Sugarcane tops and leaves have a high moisture content (70%), making them prone to mold and spoilage during transportation and storage, severely limiting their application as animal feed. To improve the utilization rate of sugarcane tops and leaves, current technology typically involves adding appropriate amounts of lactic acid bacteria, yeast, molasses, white-rot fungi, *Trichoderma viride*, and *Bacillus subtilis* to ferment protein feed after harvesting. However, these feeds still need to be used quickly, otherwise they easily spoil, making it difficult to meet the increasing demands of the current feed market. Another method involves drying or baking fresh sugarcane tops and leaves to a moisture content of 15%–20% before processing them into roughage or feed ingredients. However, due to the high lignin content of sugarcane tops and leaves, dehydration significantly increases their hardness, resulting in poor palatability, and the drying process is costly, so this method is not widely used. Sugarcane needs to be processed or sold as soon as possible after harvesting; otherwise, it is prone to moisture loss, sugar conversion, increased reducing sugars, infection by pathogens, rotting and mold growth, and a decline in flavor and taste. Currently, production mainly uses lime water spraying or chemical fungicides for preservation.

[0004] Microbial preservatives are derived from microorganisms or their metabolites. They are non-toxic and harmless, and are an important type of preservative. Screening suitable microorganisms can effectively preserve the corresponding materials. Based on this, developing microbial preservatives for sugarcane tops and leaves and sugarcane stems is an important research direction at present. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides the application of Streptomyces alogie in sugarcane preservation. Streptomyces alogie can slow down the loss of chlorophyll, water, and nutrients in sugarcane shoots and leaves after harvesting, or alleviate the rate of sugarcane stem decay and deterioration during transportation and storage, thus extending the transportation and storage period of sugarcane and its shoots and leaves. The effect is excellent and it is harmless to humans and animals.

[0006] To achieve the above objectives, the present invention provides the following technical solution: The application of Streptomyces alogie in sugarcane preservation, wherein the preservation number of Streptomyces alogie is CGMCC No: 24639, and the application is to use Streptomyces alogie to slow down the loss of chlorophyll, water and nutrients in sugarcane tops and leaves after harvesting and / or alleviate the rate of sugarcane stem decay and deterioration, and reduce the loss of water and nutrients in sugarcane stems.

[0007] Preferably, the application method is to treat sugarcane tops and leaves or sugarcane stems with Streptomyces alogiensis fermentation broth.

[0008] Preferably, the method for preparing the *Streptomyces alogiensis* fermentation broth includes the following steps: S1. Activation: Streptomyces alogie were inoculated into ISP2 solid medium for activation culture to obtain activated strains; S2, Liquid Seed Fermentation: The activated strain obtained in step S1 is inoculated into ISP2 liquid medium for culture to obtain liquid seeds; S3. Inoculate the liquid seed obtained in step S2 into ISP2 fermentation medium for fermentation culture, ensuring a viable cell count of not less than 2 × 10⁻⁶. 7 The concentration of Streptomyces alogieense was increased to 1 / mL to obtain the fermentation broth.

[0009] Preferably, the specific method for treating sugarcane tops or stems with Streptomyces alogie fermentation broth is to soak the sugarcane tops or stems in Streptomyces alogie fermentation broth for 3-5 minutes.

[0010] A sugarcane preservative containing Streptomyces alogenes as described in claim 1.

[0011] The sugarcane preservatives mentioned above can be used for preserving crop silage, food preservatives, freshness agents, and feed additives.

[0012] Preferably, the crop is sugarcane tops and leaves or sugarcane stems.

[0013] This invention provides the application of *Streptomyces alogie* in sugarcane preservation, and its advantages compared with existing technologies are: This invention verifies that *Streptomyces alogie* isolated from sugarcane leaves in Yunnan Province can effectively inhibit the growth of various pathogenic fungi, produce multiple antagonistic substances and metabolites, and is safe. It also possesses water-retaining and freshness-locking effects, showing promising development prospects in the preparation of sugarcane top and leaf silage, food preservatives, freshness-preserving agents, and feed additives. After sugarcane harvesting, soaking sugarcane stalks or top leaves with the preservative of this invention can eliminate pathogens on the plant surface, slow down the loss of chlorophyll, water, and nutrients, delay decay, preserve the plant's color, taste, flavor, and nutritional components, and extend the transportation and storage periods. The method described in this invention extends the preservation of sugarcane top leaves to 10 days and sugarcane stalks to 30 days, improving the utilization efficiency of sugarcane top and leaf resources, increasing the income of sugarcane farmers and sugar mills, and promoting the high-quality development of the sugarcane industry. Attached Figure Description

[0014] Figure 1 This is a schematic diagram comparing the preservation effects of different treatment methods on fresh sugarcane leaf fragments in embodiments of the present invention; Figure 2 This is a schematic diagram comparing the preservation effects of different treatment methods on harvested sugarcane tops and leaves in embodiments of the present invention; Figure 3 This is a schematic diagram showing the preservation effect of different treatment methods on harvested sugarcane stalks in the embodiments of the present invention. Detailed Implementation

[0015] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0016] The following Streptomyces araujoniae was deposited on April 6, 2022, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No: 24639. It is recommended to classify it as Streptomyces araujoniae and the strain number BC1.

[0017] The formulation of ISP2 solid culture medium is as follows: 10.0 g / L malt extract + 4.0 g / L glucose + 4.0 g / L yeast extract + 15 g / L agar + water (pH 7.0-7.5). After preparation, autoclave at 121℃ for 15 min. The formulations for both ISP2 liquid medium and ISP2 fermentation medium are: 10.0 g / L malt extract + 4.0 g / L glucose + 4.0 g / L yeast extract + water (pH 7.0-7.5). After preparation, autoclave at 121℃ for 15 min. Example 1:

[0018] 1. Preparation of Streptomyces alogiensis BC1 fermentation broth: After culturing Streptomyces alogie strain BC1 on ISP2 medium for 6 days, the mycelia and spores were inoculated into ISP2 liquid medium and cultured on a shaker at 30°C and 250 r / min for 48 h to obtain liquid seed. The liquid seed was then inoculated into the fermentation medium at a volume ratio of 10% and cultured at 30°C, 280 r / min, dissolved oxygen controlled at 30% volume ratio, pH 7.2, and aerated for 72 h to obtain BC1 fermentation broth.

[0019] 2. Determination of the preservation effect on fresh sugarcane leaf fragments: 2.1 Preservation Experiment Treatment: Freshly harvested sugarcane leaf fragments (Yuetang 93-159) were immersed in BC1 fermentation broth for 3 minutes and then placed in a plastic bag. The control group was treated with 75% alcohol for 3 minutes and then stored in a plastic bag.

[0020] 2.2 Observation and Testing: After 10 days, observe the color change of the sugarcane leaves, and calculate the water loss rate of the sugarcane leaves by measuring the weight difference before and after storage. The counting formula is as follows: Water loss rate (%) = (mass before storage - mass after storage) / mass before storage × 100% The specific results are shown in Table 1 below: Table 1

[0021] The results in Table 1 show that the sugarcane leaves treated with BC1 fermentation broth remained fresh, while the sugarcane leaves treated with 75% alcohol disinfection showed signs of water loss, chlorosis, and yellowing. This indicates that the supernatant from the BC1 fermentation broth can slow down the loss of chlorophyll and water from sugarcane tops and leaves after harvesting, and has a good preservation effect on sugarcane tops and leaves. Figure 1 ).

[0022] 3. Determination of the preservation effect of sugarcane tops and leaves after harvesting 3.1 Preservation Experiment Treatment: After harvesting, whole fresh sugarcane tops and leaves (Yuetang 93-159) were immersed in BC1 fermentation liquid, 75% alcohol (control group), and water (blank control) for 5 minutes, respectively. They were then stored indoors, and the color change of the sugarcane leaves was observed after 10 days.

[0023] The specific results are shown in Table 2 below: Table 2

[0024] The results in Table 2 show that the sugarcane tops and leaves treated with the BC1 fermentation broth remained green, with no reddening or rotting of the midrib; the sugarcane tops and leaves disinfected with 75% alcohol turned yellow and withered, with a few reddening and rotting of the midrib; while the sugarcane tops and leaves in the water control group turned yellow and withered, with many reddening and rotting of the midrib. This indicates that the BC1 fermentation broth can slow down the loss of chlorophyll in the sugarcane tops and leaves after harvesting and the rotting of the leaves, and has antioxidant function, thus having a good preservation effect on the sugarcane tops and leaves. Figure 2 ).

[0025] 4. Determination of the preservation effect of sugarcane stalks after harvesting: 4.1 Preservation Experiment Treatment: After harvesting, whole fresh sugarcane stalks (Yuetang 93-159) were soaked in BC1 fermentation liquid, 800 times diluted chlorothalonil (a control agent), and water (a blank control) for 3 minutes. They were then piled up indoors and cut open longitudinally with a knife after 30 days to observe the internal condition of the sugarcane stalks.

[0026] Table 3

[0027] The results in Table 3 above show that the sugarcane stalks treated with BC1 fermentation liquid remained fresh and without a bitter core. The sugarcane stalks treated with 800x dilution of chlorothalonil showed slight internal rot and no bitter core. In contrast, the sugarcane stalks treated with water showed complete internal rot, an alcoholic smell, severe water loss, and a bitter core. This indicates that BC1 fermentation liquid can effectively preserve sugarcane stalks for a long time, slowing down spoilage during transportation and storage, reducing the loss of moisture and nutrients, and extending the storage period to 30 days, thus achieving the goal of sugarcane preservation. Figure 3 ).

[0028] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Streptomyces alogius ( Streptomyces araujoniae Its application in sugarcane preservation is characterized by: The preservation number of the *Streptomyces alogie* is CGMCC No: 24639, and the application is to use *Streptomyces alogie* to slow down the loss of chlorophyll, water and nutrients in sugarcane shoots and leaves after harvesting and / or to alleviate the rate of sugarcane stem decay and deterioration, and reduce the loss of water and nutrients in sugarcane stems.

2. The application according to claim 1, characterized in that: The application method involves treating sugarcane tops and leaves or sugarcane stems with Streptomyces alogius fermentation broth.

3. The application according to claim 2, characterized in that: The preparation method of the Streptomyces alogiensis fermentation broth includes the following steps: S1. Activation: Streptomyces alogie were inoculated into ISP2 solid medium for activation culture to obtain activated strains; S2, Liquid Seed Fermentation: The activated strain obtained in step S1 is inoculated into ISP2 liquid medium for culture to obtain liquid seeds; S3. Inoculate the liquid seed obtained in step S2 into ISP2 fermentation medium for fermentation culture, ensuring a viable cell count of not less than 2 × 10⁻⁶. 7 The concentration of Streptomyces alogieense was increased to 1 / mL to obtain the fermentation broth.

4. The application according to claim 2, characterized in that: The specific method for treating sugarcane tops or stems with Streptomyces alogiensis fermentation broth is to soak the sugarcane tops or stems in Streptomyces alogiensis fermentation broth for 3-5 minutes.

5. A sugarcane preservative, characterized in that: The sugarcane preservative contains Streptomyces alogenes as described in claim 1.

6. The application of the sugarcane preservative as described in claim 5 in the preservation of crop silage, food preservative, freshness agent, and feed additive.

7. The application according to claim 6, characterized in that: The crop in question is sugarcane tops and leaves or sugarcane stems.

Images