CN116536189A - Lactic acid bacteria preparation capable of improving output and quality of wolfberry fruits and application thereof - Google Patents
Lactic acid bacteria preparation capable of improving output and quality of wolfberry fruits and application thereof Download PDFInfo
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- CN116536189A CN116536189A CN202310394806.2A CN202310394806A CN116536189A CN 116536189 A CN116536189 A CN 116536189A CN 202310394806 A CN202310394806 A CN 202310394806A CN 116536189 A CN116536189 A CN 116536189A
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- lactic acid
- lactobacillus
- acid bacteria
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 122
- 238000002360 preparation method Methods 0.000 title claims abstract description 96
- 241000894006 Bacteria Species 0.000 title claims abstract description 82
- 239000004310 lactic acid Substances 0.000 title claims abstract description 61
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 61
- 244000241838 Lycium barbarum Species 0.000 title claims description 34
- 235000015459 Lycium barbarum Nutrition 0.000 title claims description 33
- 235000015468 Lycium chinense Nutrition 0.000 title claims description 25
- 235000013399 edible fruits Nutrition 0.000 title claims description 21
- 241000186660 Lactobacillus Species 0.000 claims abstract description 53
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 53
- 235000017784 Mespilus germanica Nutrition 0.000 claims abstract description 48
- 235000000560 Mimusops elengi Nutrition 0.000 claims abstract description 46
- 235000007837 Vangueria infausta Nutrition 0.000 claims abstract description 46
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 28
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 28
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 28
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 claims abstract description 23
- 229940009289 bifidobacterium lactis Drugs 0.000 claims abstract description 23
- 241000191998 Pediococcus acidilactici Species 0.000 claims abstract description 21
- 230000001580 bacterial effect Effects 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 241000186606 Lactobacillus gasseri Species 0.000 claims description 21
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 238000009630 liquid culture Methods 0.000 claims description 6
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- 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
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Plant Pathology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Environmental Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Organic Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Agronomy & Crop Science (AREA)
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Abstract
The invention discloses a lactic acid bacteria preparation capable of improving the output and quality of medlar and application thereof, wherein active bacteria in the lactic acid bacteria preparation are at least one of bifidobacterium lactis V9, lactobacillus plantarum P-8, lactobacillus grignard G098 and pediococcus acidilactici ZPA 017. After the bacterial liquid of the lactobacillus preparation is sprayed on the root of the medlar plant, the results of related detection such as soluble protein, proline, peroxidase and the like show that the lactobacillus preparation can improve the plant resistance and adaptability, create good growth environment conditions for medlar growth, lay a solid foundation for high yield, stable yield and high quality, and greatly improve the yield and quality of medlar with the lactobacillus preparation applied.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a lactic acid bacteria preparation capable of improving the yield and quality of medlar and application thereof.
Background
The Chinese wolfberry belongs to the genus of the Solanaceae, is an important rare traditional Chinese medicine, and the perennial fallen leaves shrubs have the characteristics of drought resistance, salt and alkali resistance, high temperature resistance and barren resistance, and the Chinese wolfberry is produced by applying a large amount of chemical fertilizer to maintain the output of the Chinese wolfberry due to the long growth period, large growth quantity and large demand for mineral elements. Although the traditional farmyard manure can make up for the defect of single fertilizer application to a certain extent, the problems of low fertilizer efficiency, instability and the like still exist. However, chemical fertilizers, especially nitrogen fertilizers, are too much in application amount, which is very easy to cause the problems of product quality reduction, too high drug residue, environmental pollution and the like. With increasing importance of environmental protection and increasing ecological consciousness, microbial fertilizer gradually becomes one of important fertilizers for modern agriculture and forestry production according to the principle that biological nitrogen fixation and beneficial microorganisms can improve fertility conditions, and plays a great role in future agriculture and forestry production.
Lactic acid bacteria are bacteria which are utilized in large amounts by food microorganisms and are widely distributed in nature. A large number of researches show that lactic acid bacteria can regulate normal flora of gastrointestinal tract of an organism, maintain microecological balance, improve food digestibility and biological value, reduce serum cholesterol, control endotoxin, inhibit growth and reproduction of putrefying bacteria in the intestinal tract and generation of putrefying products, produce nutrient substances, stimulate tissue development, thereby having effects on nutritional status, physiological functions, cell infection, drug effect, toxic reaction, immune reaction, tumorigenesis, aging process, sudden emergency reaction and the like of the organism. It is well known that messages in which lactic acid bacteria play a practical role in food have become commonplace. With the continuous development of science and technology, lactic acid bacteria are increasingly used. Some lactic acid bacteria are used in dry meat products, are popularized and used due to the safety of the lactic acid bacteria, and play a great role.
In addition, some lactic acid bacteria can produce antibacterial substances, and most antibacterial spectra are relatively narrow. The bacteriocins produced by the identified lactic acid bacteria are classified into two types, one being a lactic acid bacteria having a narrow antibacterial spectrum having an inhibitory effect on only the related bacteria, and the second being a lactic acid bacteria having a broad antibacterial activity, which have an inhibitory effect on pathogenic bacteria such as clostridium botulinum, listeria and the like. Through searching, no document report for improving the quality and the yield of the medlar by adopting lactobacillus and metabolites thereof is available at present.
Disclosure of Invention
In view of the defects of the prior art, the invention provides a lactic acid bacteria preparation capable of improving the output and quality of medlar and application thereof.
In order to achieve the technical aim, the inventor combines years of research experience on various lactic acid bacteria, and passes through a large number of lactobacillus preparation consisting of a test sieve (Lactobacillus plantarum) P-8, lactobacillus gasseri G098 and Pediococcus acidilactici ZPA 017. After the bacterial liquid of the lactobacillus preparation is sprayed on the root of the medlar plant, the result shows that the medlar yield and quality are greatly improved. The technical scheme provided by the invention is as follows:
the lactobacillus preparation capable of improving the output and quality of the medlar is characterized in that active bacteria in the lactobacillus preparation are at least one of bifidobacterium lactis V9, lactobacillus plantarum P-8, lactobacillus grignard G098 and pediococcus acidilactici ZPA 017.
The bifidobacterium lactis V9 is separated from intestinal tracts of children of healthy Mongolia on inner Mongolia grasslands, has good acid resistance and bile salt resistance, and has the functions of regulating intestinal flora balance, antagonizing pathogenic bacteria in human bodies, preventing diarrhea and enhancing immunity. The strain is preserved in China general microbiological culture Collection center (CGMCC) at 11 and 18 months in 2011, and has a microorganism preservation number of CGMCC No.5470.
The lactobacillus plantarum P-8 adopted by the invention is separated from the traditional natural fermentation yoghurt of inner Mongolia, has excellent gastrointestinal digestive juice resistance and can survive in animal intestinal tracts; the strain is preserved in China general microbiological culture Collection center (CGMCC) with a preservation time of 2012, 06 and 28 days and a preservation number of CGMCC No.6312.
The lactobacillus gasseri G098 adopted by the invention is a strain separated from intestinal tracts of healthy infants in the Lassa city of the Tibetan autonomous region, and the strain shows good tolerance to artificial gastrointestinal fluids and bile salts and is a potential probiotic strain. The strain has been deposited in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) for 7 and 16 days in 2021, and has a deposit number of: cgmccno.22909.
The pediococcus acidilactici ZPA017 adopted by the invention is a pediococcus acidilactici strain with excellent performance screened from healthy piglets, and the strain has the characteristics of acid resistance, bile salt resistance, strong adhesion and good bacteriostasis. The lactobacillus strain is preserved in China general microbiological culture Collection center (North West Lu No. 1, north China center for China, the university of the science center of China, microbiological study, postal code: 100101) at the 14 th 2014, 04 th, and has the strain name Pediococcus acidilactici (Pediococcus acidilactici) ZPA017 with the preservation number: cgmccno.9040.
In one embodiment, the active bacteria in the lactobacillus preparation are the bifidobacterium lactis V9 and the lactobacillus plantarum P-8, and the ratio of the viable bacteria number of the bifidobacterium lactis V9 to the lactobacillus plantarum P-8 is (3-5): 1.
in another embodiment, the active bacteria in the lactobacillus preparation are the bifidobacterium lactis V9 and the lactobacillus gasseri G098, and the viable bacteria number ratio of the bifidobacterium lactis V9 to the lactobacillus gasseri G098 is (2-9): 1
The purpose of the invention is realized in the following way: a lactobacillus preparation capable of improving the output and quality of medlar is prepared by the following method:
(1) Inoculating at least one of the Lactobacillus bifidus V9, the Lactobacillus plantarum P-8, the Lactobacillus gasseri G098 and the Pediococcus acidilactici ZPA017 into MRS liquid culture medium, culturing for 24-35 hours at 39+ -1deg.C and pH value of 5.4-5.0, centrifuging, washing, adding 10% -12% of sterilizing degreasing Niu Ruye on thallus precipitate, and adjusting the bacterial count to 1.9-2.3X10 10 cfu/mL, mixing uniformly, pouring and culturing;
(2) Inoculating the strain cultures obtained in the step (1) into MRS culture solution respectively,culturing at 39+ -1deg.C for 30-36 hr, inoculating 10-12% of sterilized defatted Niu Ruye, culturing at 39+ -1deg.C for 10-18 hr, and regulating viable count to 3.5-5.4X10 10 cfu/mL, pH 6.5.
Further, the MRS liquid culture medium is prepared according to the following formula: the following components were dissolved in 1 liter of distilled water: 10 g of peptone, 10 g of beef extract, 5 g of yeast extract, 2 g of dipotassium hydrogen phosphate, 2 g of diammonium hydrogen citrate, 5 g of sodium acetate, 20 g of glucose, 80 ml of tween, 0.58 g of magnesium sulfate, 0.25 g of manganese sulfate, 18.0 g of agar powder, and adjusting the pH value to 6.2-6.6, and sterilizing under high pressure.
Meanwhile, the invention also provides application of the lactobacillus preparation in improving the output and quality of medlar.
Further, the lactobacillus preparation is prepared into a bacterial liquid and then poured and applied to the root of the medlar plant.
Further, the viable bacteria concentration of the bacterial liquid is 4×10 8 ～5×10 9 cfu/mL。
Further, the pH of the bacterial liquid was 6.5.
Compared with the prior art, the invention has the beneficial effects that:
compared with the prior art, the lactic acid bacteria preparation can improve the activity of protective enzymes in medlar leaves, effectively inhibit peroxidation of membranous membranes, prevent cytoplasms from being excessively damaged, thereby preventing premature senility of the leaves, improving plant resistance and adaptability, creating good growth environment conditions for medlar growth, laying a solid foundation for high yield, stable yield and high quality, and greatly improving the yield and quality of medlar applying the lactic acid bacteria preparation. The lactobacillus preparation can improve the growth and development conditions of crops, reduce the incidence rate and can be widely applied to medlar crops. Meanwhile, the lactobacillus preparation is also one of important production data for producing pollution-free foods and organic foods, and is worthy of large-area popularization and application.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the embodiments of the present application. It will be apparent that the embodiments described below are some, but not all, of the embodiments of the present application. The components of the embodiments of the present application, as generally described and illustrated herein, may be arranged and designed in a wide variety of different configurations.
Accordingly, the detailed description of the embodiments of the present application provided below in connection with the present application is intended to be merely representative of selected embodiments of the present application and is not intended to limit the scope of the claims of the present application. All other embodiments, which can be made by one of ordinary skill in the art without undue burden from the present disclosure, are within the scope of the present disclosure.
Example 1: preparation of lactic acid bacteria preparation
Preparation of MRS liquid Medium: the following components were dissolved in 1 liter of distilled water: 10 g of peptone, 10 g of beef extract, 5 g of yeast extract, 2 g of dipotassium hydrogen phosphate, 2 g of diammonium hydrogen citrate, 5 g of sodium acetate, 20 g of glucose, 80 ml of tween, 0.58 g of magnesium sulfate, 0.25 g of manganese sulfate and 18.0 g of agar powder. The pH value is adjusted to 6.2-6.6, and the mixture is sterilized under high pressure (101 Kpa,121 ℃) for 15min.
2. Preparing a skim milk culture medium: recovering skim milk powder into skim milk at 11%, naturally pH, and sterilizing at 115deg.C for 20 min.
3. Activating strain and preparing fermented milk:
(1) Inoculating Lactobacillus bifidus V9, lactobacillus plantarum P-8, lactobacillus gasseri G098 and Pediococcus acidilactici ZPA017 into MRS liquid culture medium according to at least one inoculum size, culturing at 39+ -1deg.C and pH of 5.4-5.0 for 24-35 hr, centrifuging, washing, adding 10% -12% of sterilized defatted Niu Ruye on thallus precipitate, and regulating bacterial count to 1.9-2.3X10 10 cfu/mL, mixing uniformly, pouring and culturing;
(2) The strain cultures obtained in the step (1) are respectively inoculated into MRS culture solution, cultured for 30-36 hours at 39+/-1 ℃, respectively inoculated with 10% -12% of sterilization degreasing Niu Ruye, and cultured for 10-18 hours at 39+/-1 ℃ to adjust the number of living bacteria to 3.5-5.4×10 10 cfu/mL, pH 6.5.
4. Determination of inoculum size:
the size of the inoculum size in the fermentation process can influence the growth and propagation speed of the strain. The inoculation of a small amount can lead to slow growth of thalli in the earlier stage of fermentation, prolong the fermentation period and increase the chance of bacteria contamination. The inoculation of a large amount consumes power and increases cost, the delay period of strains can be shortened, and too large inoculation amount easily causes too fast growth of thalli and causes lack of matrix nutrition. Thus, well controlled inoculum size is a key factor in ensuring proper fermentation, and different inoculum sizes have different effects on proliferation of bifidobacterium lactis V9, lactobacillus plantarum P-8, lactobacillus gasseri G098 and Pediococcus acidilactici ZPA 017. Through multiple experiments by the inventor, the bifidobacterium lactis V9, the lactobacillus plantarum P-8, the lactobacillus gasseri G098 and the pediococcus acidilactici ZPA017 are respectively according to 1% -8%: the effect is better when the inoculation amount is inoculated, and the bacteria proliferation difference in the concentration range is not large. Thus, for ease of handling, different species were inoculated at an inoculum size of 3%.
5. Determination of the optimal initial pH
The pH affects the alteration of the plasma membrane of the microorganism and at the same time affects the dissociation of part of the nutritional intermediates in the fermentation broth, so that each microorganism has a suitable pH range. If the hydrogen ion concentration in the environment exceeds the application range of the microorganism, the vital activity of the microorganism is inhibited. To determine the optimal initial pH, lactobacillus plantarum V9, lactobacillus plantarum P-8, lactobacillus gasseri G098, and Pediococcus acidilactici ZPA017 were all inoculated together, and the number of viable bacteria after 14 hours of culture in the medium at initial pH of 5.2, 5.5, 5.7, 6.0, 6.3 was measured, respectively. The initial pH greatly influences the growth conditions of the bifidobacterium lactis V9, the lactobacillus plantarum P-8, the lactobacillus gasseri G098 and the pediococcus acidilactici ZPA017, and when the initial pH is 5.2, the viable count of the bifidobacterium lactis V9, the lactobacillus plantarum P-8, the lactobacillus gasseri G098 and the pediococcus acidilactici ZPA017 cultured for 14 hours is highest, and the initial pH is too high or too low, which is unfavorable for the growth of the lactobacillus, so that the pH5.2 can be determined as the optimal initial pH value of the bifidobacterium lactis V9, the lactobacillus plantarum P-8, the lactobacillus gasseri G098 and the pediococcus acidilactici ZPA017 cultured in the MRS liquid culture medium, and therefore the pH value in the culture process can be set to be 5.4-5.0.
The best temperature determination is carried out by inoculating all of the bifidobacterium lactis V9, the lactobacillus plantarum P-8, the lactobacillus gasseri G098 and the pediococcus acidilactici ZPA017 together, and the best culture temperature of the bifidobacterium lactici V9, the lactobacillus plantarum P-8, the lactobacillus gasseri G098 and the pediococcus acidilactici ZPA017 in the culture process is 39 ℃ through multiple experiments by the inventor.
Example 2: experiment of influence of lactic acid bacteria preparation on growth, yield and quality of Lycium barbarum
1. Test method
The test was performed at 5 to 10 months in 2022 in inner mongolian baryan, mart, glade qin pioneer town. The variety of the medlar to be tested is Mongolian medlar No. one. The lactic acid bacteria preparation was prepared as in example 1, and 6 lactic acid bacteria preparations were prepared, numbered 1-6, respectively, as shown in Table 1. Geographic nature of medlar planting: the thickness of the plough layer is more than or equal to 40 cm, the organic matter content is more than or equal to 10 g/kg, the pH value is 7.0 to 8.5, and the salt content is less than 0.6%.
Selecting a planting base with smaller physical and chemical property difference, selecting a batch of medlar plants with the same growth year, small height difference, similar diameter and similar interval, dividing the medlar plants into 7 groups with the same quantity of 1-5 medlar plants, and applying lactobacillus preparations corresponding to 1-6 to the 1-6 groups on the basis of conventional fertilization, and applying clear water to the 7 groups. Wherein the application amounts of the lactobacillus preparation and the clear water are the same each time. The corresponding lactobacillus preparation and water were applied to the roots of 7 groups of experimental matrimony vine on the same day of month 6-8 of 2022, respectively. The number of application times was about 2, and the amount of application was about 10ml each time. Then, the average value of chlorophyll, soluble protein, proline, peroxidase, malondialdehyde and other contents in the leaves of the test plants, and the diseases of medlar, soil organic matters and soil enzyme activities are measured.
TABLE 1
| Numbering device | Live strain of lactobacillus preparation |
| 1 | Lactobacillus gasseri G098 |
| 2 | Bifidobacterium lactis V9 |
| 3 | Lactobacillus plantarum P-8 |
| 4 | Pediococcus acidilactici ZPA017 |
| 5 | Lactobacillus bifidus V9 and Lactobacillus plantarum P-8 |
| 6 | Bifidobacterium lactis V9 and lactobacillus gasseri G098 |
2. Experimental results
2.1 Effect of lactic acid bacteria preparation on soluble protein content in Lycium barbarum leaves (. Mu.g/g)
After lactobacillus preparation or water is applied for about two weeks each time, the same number of leaves with the same height position are extracted from the medlar tree tested in each group, one part of each group of leaves is selected for fixation, drying and crushing, and the soluble protein content of each group of medlar leaves is measured by adopting a Coomassie brilliant blue method. The experimental process is summarized, and the quality of each group of medlar leaves is equal. The results are shown in Table 2:
TABLE 2
2.2 Effect of lactic acid bacteria preparation on proline content in Lycium barbarum leaves
The proline content was measured by an acidic ninhydrin method on a part of each of the above-mentioned extracted leaves, and the measurement results are shown in Table 3:
TABLE 3 Table 3
As can be seen from Table 3, the proline content of the Lycii folium of the preparation of 6 lactic acid bacteria was higher than that of the control, wherein the proline content of the Lycii folium of the preparation of lactic acid bacteria G098 of group 1 was 28.36. Mu.g/G, wherein the proline content of the Lycii folium of the preparation of lactic acid bacteria V9 of group 2 was 29.88. Mu.g/G, wherein the proline content of the Lycii folium of the preparation of lactic acid bacteria P-8 of group 3 was 31.57. Mu.g/G, wherein the proline content of the Lycii folium of the preparation of lactic acid bacteria of group 4 lactic acid pediococcus ZPA017 was 31.32. Mu.g/G, wherein the proline content of the Lycii folium of the preparation of lactic acid bacteria V9 of group 5 and Lactobacillus plantarum P-8 was 34.01, wherein the proline content of the Lycii folium of the preparation of lactic acid bacteria V9 of group 6 and Lactobacillus plantarum G098 was 33.16, the proline content of group 26.41. Mu.g/G, and the respective control were 7.4%, 13.25%, 28.25% higher than that of the control. The proline content in the medlar leaves to which different lactobacillus preparations are applied is also different, and the proline content in the medlar leaves to which the lactobacillus preparations of the 5 th group and the 6 th group are applied is the highest.
2.3 Effect of lactic acid bacteria preparation on Peroxidase (POD) Activity in Lycium barbarum leaves
POD is responsible for H elimination in plants 2 O 2 POD is capable of catalyzing H 2 O is decomposed into H 2 O and O 2 . It is generally considered that the increased POD activity is a phenotype of plant resistanceFormula (I). By using the guaiacol method to detect the Peroxidase (POD) activity in each group of medlar leaves, as shown in Table 4, the peroxidase activity of medlar leaves is in an ascending trend after different lactobacillus preparations are applied, the POD activity in medlar leaves in group 1 is lower, and 3 lactobacillus preparations are used for remarkably increasing the peroxidase activity of medlar leaves. Lactobacillus gasseri G098 (1), bifidobacterium lactis V9 (2), lactobacillus plantarum P-8 (3), pediococcus acidilactici ZPA017 (4), bifidobacterium lactis V9 and Lactobacillus plantarum P-8 (5), bifidobacterium lactis V9 and Lactobacillus gasseri G098 (6) increased the peroxidases activity of the wolfberry leaves by 4.40%, 30.98%, 30.31%, 20.87%, 36.6% and 27.00%, respectively.
TABLE 4 Table 4
2.4 Effect of lactic acid bacteria preparation on superoxide dismutase (SOD) Activity in Lycium barbarum leaves
Plants produce free radicals in stress, which results in enhanced lipid peroxidation, leading to membrane lipid destruction and plant injury. In the long-term evolutionary adaptation process of plants, a set of active oxygen scavenging system is formed, SOD is one of the most critical enzymes for scavenging free radicals in plants, and SOD can convert O2 into O2 and H 2 O 2 。
In this example, the effect of the lactic acid bacteria preparation on the superoxide dismutase (SOD) activity in the wolfberry leaves was examined by spectrophotometry, and as shown in table 4, the SOD activity in the wolfberry leaves to which the lactic acid bacteria preparation was applied was gradually increased, and the difference in the different lactic acid bacteria preparations was gradually increased. The SOD activity of the Lycii folium of the lactobacillus preparation to which Lactobacillus bifidus V9 and Lactobacillus plantarum P-8 were applied was strongest 1.24 times that of group 7.
2.5 Effect of lactic acid bacteria preparation on Catalase (CAT) Activity in Lycium barbarum leaves
CAT is also one of the extremely important protective enzymes in organisms and is capable of scavenging intracellular excess H 2 O 2 To maintain it at a low level, protectFilm structure. In this example, the effect of the lactic acid bacteria preparation on the Catalase (CAT) activity of wolfberry leaf blades was examined by using the guaiacol method, and as shown in Table 4, it was found from the statistical analysis: compared with each control, different lactobacillus preparations have different degrees of influence on the catalase activity of the wolfberry leaves, and the difference is obvious. CAT activity in the wolfberry leaves of groups 1-6, to which different lactic acid bacteria preparations were applied, was increased by 0.70U/(g.min), 3.46U/(g.min), 3.67U/(g.min), 2.85U/(g.min), 3.66U/(g.min), and 3.37U/(g.min), respectively, as compared with the control.
2.6 Effect of lactic acid bacteria preparation on Malondialdehyde (MDA) content in Lycium barbarum leaves
In this example, the effect of different lactobacillus formulations on Malondialdehyde (MDA) content in medlar leaves was examined by MDA assay kit (micro method), and the results are shown in Table 5, and it is seen from Table 5 that the MDA content in medlar leaves to which different lactobacillus formulations were applied in groups 1 to 6 was 6.89umol/g, 5.04 umol/g, 4.13umol/g, 5.65umol/g, 3.86umol/g and 4.04umol/g, which were lower than that of the control by 5.2%, 30.8%, 43.2%, 22.3%, 46.9% and 44.4%, respectively. The Malondialdehyde (MDA) content in the wolfberry leaves of group 5 was at a lower level, with group 2 centered.
TABLE 5
2.7 Effect of lactic acid bacteria preparation on the incidence of Lycium barbarum
The occurrence of root rot, black fruit disease, leaf spot (gray spot and mildew) and virus disease of each group of medlar in the test area was investigated, and the results are shown in table 6. As can be seen from Table 6, the occurrence rate of the root virus diseases of the 1 st group, the 3 rd group and the 4 th group is 0, and the incidence rate of the root rot of the medlar is obviously reduced after the lactobacillus preparation is applied. It is seen from Table 6 that there was a reduction in the medlar disease by the application of the lactic acid bacteria preparation.
TABLE 6
2.8 Effect of lactic acid bacteria preparation on the nutrition of Lycium barbarum leaves
The lactobacillus preparation improves the plant nutrition condition, ensures that the plant nutrition condition of the healthy growth of the plant is the basis of the yield, analyzes and judges the content of nitrogen, phosphorus, potassium, calcium and magnesium in the medlar leaves, and can judge the yield-increasing potential. As shown in Table 7, the functional leaves of the medlar with the biological fertilizer are higher than the control group in nitrogen, phosphorus, potassium, calcium and magnesium, wherein the 5 th group is increased relatively more, and the functional leaves are increased by 19.68%, 26.24%, 51.38%, 28.45% and 64.48% respectively. It is demonstrated that the application of a certain amount of lactobacillus preparation can coordinate the absorption of N and promote the absorption of P, K, ca, mg element, N, P, K is the nutrition element with the largest growth and development requirement of medlar, and the balanced supply of N, P, K nutrition is the key of fertilization. Beneficial bacteria in the lactobacillus preparation can decompose P, K indissolvable in soil into P, K compounds which can be absorbed and utilized by crops, so that the crops can be promoted to absorb nutrition, the increase of potassium and calcium is beneficial to improving stress resistance of plants, and the increase of magnesium creates a material condition for chlorophyll synthesis.
TABLE 7
2.9 influence of lactic acid bacteria preparation on the phytological Properties of Lycium barbarum
The 1 st flowering and fruiting of the medlar is on the branches formed in the last year (the habit is called as "presbyopic branches", the fruiting fruit is called as "presbyopic fruits"), the 2 nd flowering and fruiting is on the branches growing in spring, the 3 rd flowering and fruiting is on the branches growing in autumn, after the presbyopic fruits are picked, the presbyopic branches accumulate nutrients for a period of time, and new branches grow out. The fruits on the presbyopic branch and the spring seven-inch branch are classified according to time, and the fruits on the autumn seven-inch branch are called summer fruits.
The length and the thickness of the spring seven-inch fruit branch and the autumn seven-inch fruit branch are measured on the medlar trees in each group in the test area, and the color of the functional leaf is observed. In addition, 10 presbyopic fruit branches are selected from each plant, 10 functional leaves are selected from each presbyopic fruit branch at random, and the size and thickness of the leaves are measured. Under the condition that the thickness of the leaves is the same as that of other treatments, the functional leaves of the presbyopic fruit branches of the 5 th group are respectively larger than those of the 6 th group, the 5 th group, the 4 th group and the control group, which indicates that the use of microbial fertilizer is beneficial to the photosynthesis of the leaves and the accumulation of photosynthetic products. Measuring the lengths and the thicknesses of the spring seven-inch fruit branch and the autumn seven-inch fruit branch, finding that the medlar applying the biological fertilizer is generally grown and thickened; the functional leaves of the presbyopic fruit branches are dark green in color, and the leaves are enlarged and thickened.
2.10 influence of the lactic acid bacteria preparation on the number of soil microorganisms
In three periods of wolfberry, respectively taking soil with the same depth of the roots of each group of wolfberry to measure the microbial count, and the results are shown in table 8, wherein the bacterial count in the soil sample applied with the lactobacillus preparation is increased, and the bacterial count in the soil after the lactobacillus preparation of the 1 st group and the lactobacillus preparation of the 2 nd group is respectively compared with that of the control group
1.83 times and 2.13 times. Actinomycetes and fungi are fewer in number than bacteria, but the total bacteria treated by the lactobacillus preparation are higher than those of the control group.
TABLE 8
2.11 influence of lactic acid bacteria preparation on the production index of Lycium barbarum
The individual yields of the medlar applying different lactobacillus preparations are shown in table 9, and the experimental results show that the individual yields of the medlar applying the lactobacillus preparations are higher than those of the control group, wherein the individual yields of the medlar of group 5 are the highest, and the individual yields of the medlar of group 1 are the lowest.
TABLE 9
2.12 influence of lactic acid bacteria preparation on the sensory and nutritional quality index of Lycium barbarum
By evaluating the fruit shape index and nutrition of the picked Chinese wolfberry of each group, the thickness (transverse diameter) of the Chinese wolfberry pulp to which the lactobacillus preparation is applied is found to be higher than that of a control group, wherein the transverse diameter of the Chinese wolfberry pulp of the 3 rd group is 7mm at maximum. The weight of single fruit of fructus Lycii with lactobacillus preparation applied exceeds that of control group. Wherein the weight of single fruit of Lycium barbarum of group 5 is 11.35mg.
In the aspect of nutrition, the wolfberry polysaccharide content of the wolfberry which is applied with the lactobacillus preparation is higher than that of the control group by detecting the wolfberry polysaccharide of each group of the obtained wolfberry.
In summary, the invention provides the lactobacillus preparation capable of improving the yield and the quality of the medlar, compared with a control group, the lactobacillus preparation can improve the activity of protective enzymes in medlar leaves, effectively inhibit membranous peroxidation, prevent cytoplasmic membranes from being excessively damaged, thereby preventing premature senility of the leaves, improving plant resistance and adaptability, creating good growth environment conditions for medlar growth, laying a solid foundation for high yield, stable yield and high quality, and greatly improving the yield and the quality of medlar applying the lactobacillus preparation. The lactobacillus preparation can improve the growth and development conditions of crops, reduce the incidence rate and can be widely applied to medlar crops. Meanwhile, the lactobacillus preparation is also one of important production data for producing pollution-free foods and organic foods, and is worthy of large-area popularization and application.
The foregoing is merely a preferred embodiment of the present application, but the scope of the present application is not limited thereto, and any changes or substitutions within the technical scope of the present disclosure should be covered by the scope of the present application. Therefore, the protection scope of the present application shall be subject to the protection scope of the claims.
Claims (9)
1. A lactic acid bacteria preparation capable of improving the output and quality of medlar is characterized in that active bacteria in the lactic acid bacteria preparation are at least one of bifidobacterium lactis V9, lactobacillus plantarum P-8, lactobacillus grignard G098 and pediococcus acidilactici ZPA 017.
2. The lactic acid bacteria preparation according to claim 1, characterized in that:
the active bacteria in the lactobacillus preparation are the bifidobacterium lactis V9 and the lactobacillus plantarum P-8, and the ratio of the viable bacteria number of the bifidobacterium lactis V9 to the lactobacillus plantarum P-8 is (3-5): 1.
3. the lactic acid bacteria preparation according to claim 1, characterized in that:
the active bacteria in the lactobacillus preparation are the bifidobacterium lactis V9 and the lactobacillus gasseri G098, and the ratio of the viable bacteria number of the bifidobacterium lactis V9 to the lactobacillus gasseri G098 is (2-9): 1.
4. a lactic acid bacteria preparation according to any one of claims 1-3, characterized in that the lactic acid bacteria preparation is prepared according to the following method:
(1) Inoculating at least one of the Lactobacillus bifidus V9, the Lactobacillus plantarum P-8, the Lactobacillus gasseri G098 and the Pediococcus acidilactici ZPA017 into MRS liquid culture medium, culturing for 24-35 hours at 39+ -1deg.C and pH value of 5.4-5.0, centrifuging, washing, adding 10% -12% of sterilizing degreasing Niu Ruye on thallus precipitate, and adjusting the bacterial count to 1.9-2.3X10 10 cfu/mL, mixing uniformly, pouring and culturing;
(2) Inoculating the strain cultures obtained in the step (1) into MRS culture solution respectively, culturing for 30-36 hours at 39+ -1deg.C, inoculating 10% -12% of sterilized defatted Niu Ruye respectively, culturing for 10-18 hours at 39+ -1deg.C, and adjusting the viable count to 3.5-5.4X10 × 10 cfu/mL, pH 6.5.
5. The lactic acid bacteria preparation according to claim 4, characterized in that:
the MRS liquid culture medium is prepared according to the following formula: the following components were dissolved in 1 liter of distilled water: 10 g of peptone, 10 g of beef extract, 5 g of yeast extract, 2 g of dipotassium hydrogen phosphate, 2 g of diammonium hydrogen citrate, 5 g of sodium acetate, 20 g of glucose, 80 ml of tween, 0.58 g of magnesium sulfate, 0.25 g of manganese sulfate, 18.0 g of agar powder, and adjusting the pH value to 6.2-6.6, and sterilizing under high pressure.
6. Use of the lactic acid bacteria preparation of claim 4 for improving the yield and quality of wolfberry.
7. The use of the lactic acid bacteria preparation according to claim 6 for improving the yield and quality of wolfberry, wherein the lactic acid bacteria preparation is prepared into bacterial liquid and then poured on the root of wolfberry plant.
8. The use of the lactic acid bacteria preparation for improving the yield and quality of wolfberry fruit according to claim 7, wherein the viable bacteria concentration of the bacterial liquid is 4 x 10 8 ～5×10 9 cfu/mL。
9. Use of a lactic acid bacterial preparation according to claim 7 or 8 for increasing the yield and quality of wolfberry, characterized in that the PH of the bacterial liquid is 6.5.
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