Treatment method for promoting germination of asparagus cochinchinensis seeds

Abstract

The invention discloses a treatment method for promoting germination of asparagus cochinchinensis seeds, and belongs to the technical field of biology, the treatment method comprises the following steps: S1, soaking seeds: selecting mature and full fresh asparagus cochinchinensis seeds, soaking for 72-96 hours by using water at 40-60 DEG C, and fishing out for later use after soaking; s2, preparing a complex microbial inoculant: mixing a stable burkholderia liquid microbial inoculant and a bacillus licheniformis liquid microbial inoculant of which the effective viable count is not less than 1 * 10 < 8 > CFU / ml according to a volume ratio of 1: (3-5), so as to obtain the complex microbial inoculant, wherein the volume ratio of the stable burkholderia liquid microbial inoculant to the bacillus licheniformis liquid microbial inoculant is 1: (3-5); s3, germination promotion with a complex microbial inoculant: putting filter paper in a container, wetting the filter paper with the complex microbial inoculant, putting the seeds treated in the step S1 on the filter paper, and culturing at room temperature under a dark condition. According to the treatment method for germination of the asparagus cochinchinensis seeds, the germination time of the asparagus cochinchinensis seeds can be effectively shortened, and the germination rate of the asparagus cochinchinensis seeds can be effectively increased.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biotechnology, and particularly relates to a treatment method for promoting seed germination of asparagus. BACKGROUND

[0002] Radix Asparagi is a perennial climbing herb of Asparagus cochinchinensis (Lour.) Merr. in the Liliaceae family, and the dried tuber is used as medicine. Radix Asparagi is both medicine and food, and the market demand for medicinal materials is large, and the artificial planting area is increasing year by year. The seedling methods of artificially planted Radix Asparagi mainly include division planting, tissue culture breeding and seed propagation. Because the division propagation coefficient is low, the tissue culture technology and facility requirements are high, and at present, seed propagation is mostly used, but seed propagation has the problems of long germination time and low germination rate. SUMMARY

[0003] In order to overcome the problems in the prior art, the application provides a treatment method for promoting seed germination of asparagus.

[0004] To achieve the above-mentioned purpose, the application is realized by the following technical scheme: A treatment method for promoting seed germination of asparagus, comprising the following steps: S1, soaking seeds: selecting mature and full fresh asparagus seeds, soaking the seeds in water at 40-60 DEG C for 72-96 h, and then taking out the seeds for standby after soaking; S2, preparing a compound microbial agent: mixing stable Burkholderia liquid microbial agents and Bacillus licheniformis liquid microbial agents with effective viable cell counts of not less than 1x10 8 CFU / ml at a volume ratio of 1:3-5 to obtain a compound microbial agent; S3, promoting germination by the compound microbial agent: placing filter paper in a container, wetting the filter paper with the compound microbial agent, and then placing the seeds obtained after the treatment in step S1 in the dark at room temperature.

[0005] Further, the preparation method of the stable Burkholderia liquid microbial agent comprises the following steps: A1, configuring a liquid fermentation medium: the liquid fermentation medium comprises 10.0 g / L of proteose peptone, 3.0 g / L of beef powder and 5.0 g / L of sodium chloride, and the pH is adjusted to 7.2-7.4; A2, inoculating stable Burkholderia spores into the above-mentioned liquid fermentation medium, and then performing aerobic fermentation at 30 DEG C for 18-24 h to treat the fermentation liquor so that the effective viable cell count is not less than 1x10 8 CFU / ml, and the stable Burkholderia liquid microbial agent is obtained.

[0006] Further, the preparation method of the bacillus licheniformis liquid inoculant comprises the following steps: B1, configure a liquid fermentation medium: the liquid fermentation medium comprises: corn flour 5.0 g, proteose peptone 10.0 g, soybean meal 3.0 g, yeast paste 5.0 g, manganese sulfate 0.1 g, potassium dihydrogen phosphate 1 g, magnesium sulfate 0.2 g, and sterile water 1 L, and the pH is adjusted to 7.2-7.4; B2, inoculate the bacillus licheniformis strain into the above-mentioned liquid fermentation medium, and then perform aerobic fermentation at 30 DEG C for 18-24 h, and treat the fermentation liquor so that the effective viable count is not less than 1x10 8 CFU / ml, and the bacillus licheniformis liquid inoculant is obtained.

[0007] Further, the stable burkholderia number is BNCC135093, and the bacillus licheniformis number is BNCC368151, both of which are purchased from the BNCC ® Industrial Microbial Strain Engineering Technology Research Center.

[0008] Through the above technical solution, the asparagus seed germination treatment method of the present application can effectively shorten the asparagus seed germination time and improve the asparagus seed germination rate. DETAILED DESCRIPTION

[0009] Unless otherwise specified, the materials and reagents used in the present application are commercially available.

[0010] In the present application, the stable burkholderia number is BNCC135093, and the bacillus licheniformis number is BNCC368151, both of which are purchased from the BNCC ® Industrial Microbial Strain Engineering Technology Research Center.

[0011] The preparation method of the stable burkholderia liquid inoculant comprises the following steps: A1, configure a liquid fermentation medium: the liquid fermentation medium comprises: proteose peptone 10.0 g / L, beef powder 3.0 g / L, and sodium chloride 5.0 g / L, and the pH is adjusted to 7.2-7.4; A2, inoculate the stable burkholderia strain into the above-mentioned liquid fermentation medium, and then perform aerobic fermentation at 30 DEG C for 18-24 h, and treat the fermentation liquor so that the effective viable count is about 1x10 8 CFU / ml, and the stable burkholderia liquid inoculant is obtained.

[0012] The preparation method of the bacillus licheniformis liquid inoculant comprises the following steps: B1, configure liquid fermentation medium: the liquid fermentation medium comprises: corn flour 5.0g, proteose peptone 10.0g, soybean meal 3.0g, yeast paste 5.0g, manganese sulfate 0.1g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.2g, sterilized water 1L, adjust pH to 7.2-7.4; B2, after inoculating bacillus licheniformis strain into the above-mentioned liquid fermentation medium, aerobic fermentation is carried out at 30 DEG C for 18-24h, and the effective viable cell number of the fermentation liquor is about 1x10 8 CFU / ml, and bacillus licheniformis liquid inoculant is obtained.

[0013] Example one A treatment method for promoting asparagus seed germination, comprising the following steps: S1, soaking seeds: select mature and full fresh asparagus seeds, soak them in 50 DEG C water for 84h, and then take them out for standby after soaking; S2, preparing composite inoculant: mix stable burkholderia liquid inoculant and bacillus licheniformis liquid inoculant, both of which have effective viable cell number not less than 1x10 8 CFU / ml, at a volume ratio of 1:4, and then obtain the composite inoculant; S3, composite inoculant promotes germination: pad filter paper in a container, wet the filter paper with the composite inoculant, and then put the seeds obtained after S1 step treatment, and cultivate at room temperature in dark conditions.

[0014] Example two A treatment method for promoting asparagus seed germination, comprising the following steps: S1, soaking seeds: select mature and full fresh asparagus seeds, soak them in 40 DEG C water for 72h, and then take them out for standby after soaking; S2, preparing composite inoculant: mix stable burkholderia liquid inoculant and bacillus licheniformis liquid inoculant, both of which have effective viable cell number not less than 1x10 8 CFU / ml, at a volume ratio of 1:3, and then obtain the composite inoculant; S3, composite inoculant promotes germination: pad filter paper in a container, wet the filter paper with the composite inoculant, and then put the seeds obtained after S1 step treatment, and cultivate at room temperature in dark conditions.

[0015] Example three A treatment method for promoting asparagus seed germination, comprising the following steps: S1, soaking seeds: select mature and full fresh asparagus seeds, soak them in 60 DEG C water for 96h, and then take them out for standby after soaking; S2, preparing composite inoculant: mix stable burkholderia liquid inoculant and bacillus licheniformis liquid inoculant, both of which have effective viable cell number not less than 1x10 8A composite inoculum was obtained by mixing stable Burkholderia holmium liquid inoculum (CFU / ml) and Bacillus licheniformis liquid inoculum at a volume ratio of 1:5. S3, promoting germination with compound microbial agent: Place filter paper in a container, moisten the filter paper with compound microbial agent, and then place the seeds obtained after treatment in step S1 on it. Cultivate at room temperature in the dark.

[0016] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that only stable Burkholderia liquid inoculant was used to promote germination.

[0017] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that only Bacillus licheniformis liquid inoculant was used to promote germination.

[0018] Germination Experiment Mature and plump fresh asparagus seeds were collected and treated according to the technical solutions described in Examples 1 to 3 and Comparative Examples 1 to 2. Seed germination experiments were then conducted, with three replicates for each treatment and 100 seeds per treatment. Seed germination was observed daily, and the initial germination time, germination duration, and germination rate for each treatment were recorded. The statistical results are shown in Table 1 below.

[0019] Seed germination test standards The germination initiation time and germination duration of asparagus seeds were determined. A seed was considered to be germinating when the hypocotyl appeared, and the appearance of the hypocotyl of the first seed was the germination initiation time for this treatment. Germination was considered to have ended when the germination rate did not exceed 1% of the replicate number of seeds for 3 consecutive days, and the germination end time was three days prior.

[0020] Germination rate is an indicator of a seed's ability to germinate, calculated using the following formula: Germination rate (GR) = (Number of germinated seeds / 100) * % Table 1. Statistics on initial germination time, germination duration, and germination rate of asparagus seeds for each treatment.

[0021] As can be seen from Table 1 above, the treatment method for promoting the germination of asparagus seeds described in this invention can effectively shorten the germination time of asparagus seeds and increase the germination rate of asparagus seeds.

[0022] Finally, it should be noted that the above preferred 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 through the above preferred embodiments, those skilled in the art should understand that various changes can be made to it in form and detail without departing from the scope defined by the claims of the present invention.

Claims

1. A treatment method for promoting the germination of asparagus seeds, characterized in that: It includes the following steps: S1, Soaking seeds: Select mature and plump fresh asparagus seeds and soak them in water at 40-60℃ for 72-96 hours. After soaking, take them out and set them aside. S2, Preparation of compound bacterial agent: Ensure that the effective viable bacteria count is not less than 1×10⁻⁶. 8 A composite inoculum was obtained by mixing stable Burkholderia holmium liquid inoculum (CFU / ml) with Bacillus licheniformis liquid inoculum at a volume ratio of 1:3-5. S3, promoting germination with compound microbial agent: Place filter paper in a container, moisten the filter paper with compound microbial agent, and then place the seeds obtained after treatment in step S1 on it. Cultivate at room temperature in the dark.

2. The treatment method for promoting the germination of asparagus seeds according to claim 1, characterized in that: The method for preparing the stable Burkholderia liquid inoculum includes the following steps: A1. Preparation of liquid fermentation medium: The liquid fermentation medium contains: 10.0g peptone, 3.0g beef meal, 5.0g sodium chloride, 1L sterile water, and the pH is adjusted to 7.2-7.4; A2. After inoculating the stable Burkholderia strain into the above-mentioned liquid fermentation medium, aerobic fermentation was carried out at 30°C for 18-24 hours. The fermentation broth was then treated to ensure that the effective viable cell count was not less than 1×10⁻⁶. 8 CFU / ml yields a stable Burkholderia liquid inoculum.

3. The treatment method for promoting the germination of asparagus seeds according to claim 1, characterized in that: The preparation method of the Bacillus licheniformis liquid inoculant includes the following steps: B1. Preparation of liquid fermentation medium: The liquid fermentation medium contains: 5.0g corn flour, 10.0g peptone, 3.0g soybean meal, 5.0g yeast extract, 0.1g manganese sulfate, 1g potassium dihydrogen phosphate, 0.2g magnesium sulfate, 1L sterile water, and pH adjusted to 7.2-7.

4. B2. After inoculating the Bacillus licheniformis strain into the above-mentioned liquid fermentation medium, aerobic fermentation was carried out at 30℃ for 18-24 hours. The fermentation broth was then treated to ensure that the effective viable cell count was not less than 1×10⁻⁶. 8 CFU / ml yields a liquid Bacillus licheniformis inoculant.

4. The treatment method for promoting the germination of asparagus seeds according to claim 1, characterized in that: The stable Burkholderia species, catalog number BNCC135093, and Bacillus licheniformis species, catalog number BNCC368151, were both purchased from BNCC. ® Research Center for Engineering Technology of Industrial Microbial Strains

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