CN117652527A - Yang Meicheng flowering phase regulator and preparation method thereof - Google Patents
Yang Meicheng flowering phase regulator and preparation method thereof Download PDFInfo
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the field of plant growth regulators, and particularly relates to a poplar full bloom regulator and a preparation method thereof. The method comprises the following steps: 1) Preparing a culture medium, sub-packaging after inactivation, inoculating strains into one culture medium, and carrying out timed shaking culture to obtain a fermentation broth; 2) Inoculating strain into another culture medium, sealing, standing for culturing, centrifuging, filtering, and inactivating to obtain filtrate; 3) Taking out the fermentation liquor, adding the filtrate, and uniformly mixing to obtain the Yang Meicheng flowering phase regulator. The waxberry quality regulator prepared by the invention can promote soil fertility, is beneficial to the fruiting of waxberry trees in the growing period and fruiting period, especially in the full bloom period, has higher fruiting rate than the waxberry trees with the traditional fertilizer, and can promote the accumulation of soluble solids, soluble sugar and vitamin C in the waxberry fruits.
Description
Technical Field
The invention belongs to the field of plant growth regulators, and particularly relates to a poplar full bloom regulator and a preparation method thereof.
Background
In recent years, the novel debilitation disease of the waxberry tree causes that more than 80% of mature leaves of the waxberry tree fall off, the leaves of the whole tree body are sparse and dark in color and luster, and the roots form blackening and ulceration until the tree body dies after 2-4 years, and the like, so that the yield and the quality of the waxberry are influenced by different degrees, and the healthy development of the industry is greatly restricted.
The technical staff can improve a series of problems caused by the novel debilitating symptoms of the waxberries by improving soil environments such as soil pH value, microbial community imbalance and the like. The application of the nitrogen and phosphorus compound fertilizer can effectively promote the growth of crops, further increase the nutrition components of the fertilizer, increase the nutrition absorption rate of plants and further improve the yield and quality of waxberry fruits.
Patent CN 110540451a discloses a nutritional fertilizer for waxberry, which belongs to the technical field of fertilizer preparation and comprises the following materials in percentage by weight: 10 to 30 percent wt percent of sarcandra glabra, 2 to 6 percent wt percent of malic acid, 0.5 to 2.5 percent wt percent of ethylene, 1.2 to 2.6 percent wt percent of gibberellin, 3.5 to 5.5 percent wt percent of magnesium sulfate, 5 to 20 percent wt percent of tapioca flour, 1.5 to 6.5 percent wt percent of urea, 5 to 10 percent wt percent of bean cake powder, 1 to 6 percent wt percent of dispersing agent and the balance of water. As the components such as ethylene, gibberellin, magnesium sulfate and the like are added, the plant diseases and insect pests of crops can be prevented.
However, the sensitivity of the waxberry to trace elements is higher, for example, boron deficiency can lead to the sterilization of the waxberry, inhibit the growth of the tree body and even lead to the death of the plant, and excessive application of the nitrogen and phosphorus compound fertilizer can cause adverse effects on the nitrogen fixation and phosphorus dissolution functions of the symbiotic microorganisms of the root system of the waxberry, and can lead to the defoliation and even the death of the tree body when serious. Therefore, how to reasonably and dynamically regulate the nutrient environment in the growing environment of the waxberries through the synergistic effect of microorganisms so as to improve the growing development trend of the waxberries is always an important research object.
In this regard, researchers have previously studied and published related papers: hong Lei east, etc., and the special bio-organic fertilizer has the effect of improving the soil and improving the quality of the fruits of the waxberry with the debilitation. In the paper, the application of the special microbial inoculum in cooperation with silkworm excrement organic fertilizer in the late middle and late ten days of July and the early spring tip of December is studied in depth, and then the soil index and the fruit quality are characterized, wherein the characterization result shows that the soil index and the fruit quality show effectiveness, but in the further study, the effect generated after the special microbial inoculum is applied in different periods is found to have variability, and the specific application mode also influences the specific application effect to a certain extent.
Therefore, the invention further researches the preparation and application of the microbial inoculum composite regulator.
Disclosure of Invention
The invention provides a Yang Meicheng flowering phase regulator, a preparation method and specific application thereof, and aims to solve the problems that the existing regulator is difficult to improve fallen leaves and yield reduction caused by novel debilitation of waxberry trees, and the quality of fruits is reduced, and the use effect of the novel microbial agent is still to be improved.
The invention aims at:
1. the regulator can effectively improve the symptoms of the novel debilitation of the waxberry tree;
2. the regulator can effectively regulate the fruit development of the waxberry tree.
In order to achieve the above purpose, the present invention adopts the following technical scheme.
A preparation method of Yang Meicheng flowering phase regulator,
the method comprises the following steps:
1) Preparing a culture medium, sub-packaging after inactivation, inoculating strains into one culture medium, and carrying out timed shaking culture to obtain a fermentation broth;
2) Inoculating strain into another culture medium, sealing, standing for culturing, centrifuging, filtering, and inactivating to obtain filtrate;
3) Taking out the fermentation liquor, adding the filtrate, and uniformly mixing to obtain the Yang Meicheng flowering phase regulator.
As a preferred alternative to this,
the culture medium in the step 1) is a molasses culture medium.
As a preferred alternative to this,
the strains in the step 1) are microbacterium phyllosum and greedy bacteria;
the inoculation ratio of the microbacterium phyllosum to the greedy bacteria is 1: (0.9-1.1).
As a preferred alternative to this,
the concentration of bacteria in the fermentation liquor obtained in the step 1) is 1 multiplied by 10 9 ～1×10 10 CFU/mL。
As a preferred alternative to this,
the strain in the step 2) is photosynthetic bacteria rhodopseudomonas palustris, and the culture is carried out in a sealed static culture process until the concentration of the photosynthetic bacteria rhodopseudomonas palustris is 1 multiplied by 10 9 ～1×10 10 CFU/mL。
As a preferred alternative to this,
the dosage volume ratio of the fermentation liquor to the filtrate in the step 3) is 9: (0.9-1.1).
A Yang Meicheng flowering phase regulator.
As a preferred alternative to this,
when the Yang Meicheng flowering phase regulator is used, the Yang Meicheng flowering phase regulator and a microbial carrier are mixed according to the proportion of 0.008-0.012: mixing the materials according to the mass ratio of 1;
the microbial carrier is an organic fertilizer.
As a preferred alternative to this,
the Yang Meicheng flowering phase regulator is applied in a loop according to the dosage ratio of 190-220 g/tree of Myrica rubra 1-3 w before Yang Meicheng flowering phase.
In the technical scheme of the invention, the most important is that the fermentation liquor is prepared by utilizing microbacterium phyllosum and greevovorax (Variovorax), endophytes and saprophytes generated in the waxberry tree are effectively inhibited, and the cultivation of the waxberry tree is promoted by adopting a mode of the synergistic action of microorganism strains and soil microorganism communities, so that the root system development is obviously promoted, in particular the vertical development of the root system is promoted, the disease resistance of the sapling is effectively improved, and the survival rate of the sapling is improved. According to the invention, the elements N, P, K and the like in the soil are fixed by utilizing the mycelia of the two elements, so that the elements are converted into nutrition which can be directly absorbed and utilized by the myrica rubra, a good environment is provided for the myrica rubra, and the effect of promoting root system growth and development is achieved. Wherein the Bacillus phyllovis produces an antibacterial substance with a purine structure in the fermentation process, and can form dominant bacterial groups in soil when in use, thereby inhibiting the growth of harmful bacterial groups, simultaneously, a large amount of extracellular polysaccharide and small molecular protein are generated in the growth process, so that the tree can be planted at the root tip of the tree body, and the extracellular space outside the root dimension column is accumulated, thereby promoting nitrogen fixation of the Frankia actinomycetes and accelerating the process of absorbing nutrient substances by the tree body. The quality and maturity of fruits can be effectively regulated and controlled by the greedy bacteria, for example, fructose and glucose in waxberry can influence sweetness of the fruits, sucrose with higher concentration can serve as a protective agent, oxidase is activated, cold resistance of the fruits is enhanced, and therefore survival rate of the tree is improved.
Further, the invention adds the fermentation filtrate of photosynthetic bacteria rhodopseudomonas palustris (Rhodopseudomonas palustris) into the fermentation liquor prepared by the microbacterium phyllosum and the greedy bacteria, and the photosynthetic bacteria rhodopseudomonas palustris metabolized to produce substrates, intermediate reactants, energy, reducing substances and the like. The metabolic active substances (cytokinin and auxin) accumulate in the soil, stimulate the growth of Frankia actinomycetes in the soil, reduce the quantity of fungi and fusarium oxysporum, improve the activities of enzymes such as neutral phosphatase and sucrase, improve the survival rate of trees and improve the sugar content of fruits. Therefore, the growing environment of the flora is balanced, the continuous propagation of dominant flora is facilitated, and the quality of fruits is further improved.
Based on the special composite microbial inoculum regulator, the invention further researches the use time machine.
Myrica rubra is a plant of hermaphroditic plants. Flowering and leaf expansion are both in spring, and the growing points are between axils at the upper part of branch tips. Flowering is performed first and then the tip is pulled out to expand leaves. The male flowers are opened from the last 2 months to the last 4 months, and the last 3 months are full-bloom periods, and the bloom periods are 1.5 months long; estrus is blooming from 3 months up to 4 months, and the flowering period is about 20 days longer in 3 months.
The full bloom stage refers to the full bloom stage of the female tuo, namely the full bloom stage of the male flowers is 1-2 week earlier than the time point, so that the application time is usually the time after the male flowers enter the full bloom stage. The full bloom stage of the myrica rubra is a key period of bud expansion and flowering, and in the key period, the myrica rubra needs to accumulate a large amount of nutrients and carry out reasonable and efficient transportation and utilization on the myrica rubra. In general, fertilization at this stage requires strict control of fertilization amount, fertilization period and monitoring of nutrient content of soil, and in combination with effective pruning, the pruning process adopts a method mainly comprising thinning and puncturing, proper puncturing, thinning and removing excessive dense branches, upright branches and weakened and lengthy branches of the shortened bore, and updating the branch group. This is also mainly due to uneven nutrient supply, limited nutrient utilization of the myrica rubra, limited development of a large number of fruiting branches, and easy nutrient competition of dense branches. In this regard, the invention can realize the acceleration of the absorption of the waxberry tree to the absorbable nutrient of the soil through the regulation and control of the soil microenvironment, and on the basis, the invention can not only strengthen the root system of the waxberry tree and make the waxberry tree quickly absorb the nutrient substances, but also improve the rate of the waxberry tree for conveying the nutrient substances to the branch tips, thereby greatly improving the nutrient utilization level of the waxberry tree and further improving the development effect of the waxberry tree.
The beneficial effects of the invention are as follows:
(1) The waxberry quality regulator prepared by the invention can promote soil fertility, is beneficial to fruit bearing of waxberry trees in a growing period and a fruiting period, and is especially applied to a full bloom period, and the fruit bearing rate of the waxberry trees is higher than that of waxberry trees applying traditional fertilizers;
(2) The waxberry quality regulator prepared by the invention can promote the accumulation of soluble solids, soluble sugar and vitamin C in waxberry fruits;
(3) The waxberry quality regulator prepared by the invention can effectively improve the enzyme activity of single waxberry fruits, and the obtained single waxberry fruits are larger in size, plump and high in quality.
Detailed Description
The present invention will be described in further detail with reference to specific examples. Those of ordinary skill in the art will be able to implement the invention based on these descriptions. In addition, the embodiments of the present invention referred to in the following description are typically only some, but not all, embodiments of the present invention. Therefore, all other embodiments, which can be made by one of ordinary skill in the art without undue burden, are intended to be within the scope of the present invention, based on the embodiments of the present invention.
The raw materials used in the examples of the present invention are all commercially available or available to those skilled in the art unless specifically stated otherwise; the methods used in the examples of the present invention are those known to those skilled in the art unless specifically stated otherwise.
The strain sources used in the embodiment of the invention are CGMCC (China center for culture collection of microorganisms) unless specified.
The honey culture medium used in the embodiment of the invention is a conventional honey culture medium, and the honey culture medium comprises the following components: cane molasses 10 g, wheat bran 6 g, corn flour 2 g, K 2 HPO 4 2g，MnSO 4 ·H 2 O 0.05 g，MgSO 4 ·7H 2 O0.1 g, pH 7.2, pure water 1000 mL, sterilization at 121℃for 20min.
Example 1
A Yang Meicheng flowering phase regulator prepared by the method of:
1) Preparing a molasses culture medium, inactivating the molasses culture medium at 121 ℃ for 20min, sub-packaging, and inoculating microbacterium phyllosum and greedy bacteria into one culture medium, wherein the inoculation ratio of the microbacterium phyllosum to the greedy bacteria is 1:1, culturing 48 h by time shaking to obtain fermentation brothThe concentration of bacteria in the fermentation broth is about 9.2×10 9 CFU/mL；
2) Inoculating rhodopseudomonas palustris as photosynthetic bacteria into the other culture medium, and culturing under sealed condition 48. 48 h until the bacterial concentration is about 8.1X10% 9 CFU/mL, centrifuging, filtering and ultraviolet inactivating to obtain filtrate;
3) Taking out the fermentation liquor, and mixing the fermentation liquor and filtrate according to the volume ratio of the usage amount of 9: and 1, adding the filtrate in proportion, and uniformly mixing to obtain the Yang Meicheng flowering phase regulator.
The waxberry tree with the novel debilitation is used as a test sample. The Yang Meicheng flowering phase regulator prepared by the method and silkworm excrement organic fertilizer are mixed according to the mass ratio of 0.01:1, and evenly and annularly applying the mixture according to the dosage ratio of 2000 g/myrica rubra tree after the male flowers enter the full bloom stage in the last ten days of 3 months of 2023. The experimental place is in a red bayberry orchard of Jin Huashi lanxi in Zhejiang province, and near 30 red bayberry trees which are planted in the same period, grow for 8 years and have premature senility symptoms are planted in the red bayberry orchard, and the red bayberry trees belong to Dongkui species, and have nearly 200 red bayberry trees of the same variety in total. And the other period adopts conventional fertilization. If no special description exists, the soil and the fruits are sampled before the same day of harvesting in 7 th month of the current year, and the fruit sampling is only carried out by randomly selecting 8-10 fruits on each myrica rubra tree. And in the full bloom stage, the partial pruning of the dense branches is not carried out, and at least 2 of the fruits are ensured to be dense branch fruits during fruit sampling.
The characterization specifically comprises the following steps:
1. physical and chemical properties of soil:
the pH value is measured by an acidometer, the conductivity is measured according to ISO11265:1994 (E) international standard, the organic matter content is measured by a potassium dichromate-sulfuric acid hydration heat method, the total nitrogen content is measured by a Kjeldahl nitrogen method, the effective nitrogen element content is measured by a Kjeldahl nitrogen method, the effective phosphorus element content is measured by a molybdenum-antimony colorimetric resistant method, and the effective potassium element content is measured by NH 4 Ac leaching-flame photometry. Carrying out two times of characterization on physical and chemical indexes of soil, wherein the first time is one day before fertilization, and the second time is sampling before harvesting;
2. the physical index of the waxberry is as follows:
the amino acid content adopts an ninhydrin colorimetric method, the vitamin C content adopts an iodine titration method, the flavonoid content adopts a near infrared hyperspectral method, the phenolic acid content adopts a NaOH neutralization titration method, and the soluble sugar content adopts an anthrone colorimetric method;
3. the quality index of the waxberry fruits is as follows:
measuring transverse diameter and longitudinal diameter of the waxberry fruits by a vernier caliper, measuring single fruit weight of the waxberry fruits by a balance, measuring fruit hardness by a TA-XTplus texture instrument, measuring the content of soluble solids by a handheld glycometer, and recording the average value;
4. good fruit rate:
the excellent fruit rate is characterized in that all the fruits are harvested, the surfaces of the fruits are clean and have no mildew spots, the fruit shapes are right, the scars account for less than 10% of the fruit surfaces, the excellent fruit rate is calculated, and the formula is shown as (N/N) multiplied by 100%, wherein N is the number of the excellent fruits, and N is the harvesting number.
All experimental data were analyzed using GraphPad Prism Version 10.0.2 software and the data are expressed as mean ± standard deviation. One-way analysis of variance (ANOVA) using Tukey test, with at least 3 replicates per treatment, results were as follows * Indicating that the treatment has significant differences compared to the control, i.e. P<0.05）。
The sample characterization results are as follows.
Soil sampling physical and chemical indexes one day before fertilization:
| pH | conductivity (mu s/cm) | Organic matter (g/kg) | Total nitrogen (mg/kg) | Available nitrogen (mg/kg) | Available phosphorus (mg/kg) | Effective potassium (mg/kg) |
| 4.61±0.34 | 96.85±30.03 | 34.30±6.91 | 1.36±0.32 | 119.46±30.86 | 8.69±6.60 | 133.83±9.90 |
Sampling soil sampling physical and chemical indexes before harvesting:
| pH | conductivity (mu s/cm) | Organic matter (g/kg) | Total nitrogen (mg/kg) | Available nitrogen (mg/kg) | Available phosphorus (mg/kg) | Effective potassium (mg/kg) |
| 4.89±0.30 * | 97.40±12.85 | 28.25±0.35 | 1.09±0.05 * | 104.80±4.55 | 45.00±5.57 * | 120.00±19.10 |
Non-dense branch red bayberry physiological index:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 3.057 | 1389.599 | 0.659 | 1.245 | 106.715 |
The quality index of the non-dense branch red bayberry fruits:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 32.71 | 30.92 | 18.39 | 3.57 | 12.7 | 89 |
The physiological index of the dense branch red bayberry:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.867 | 1286.113 | 0.613 | 1.222 | 98.091 |
The quality index of the thick-branch waxberry fruits is as follows:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 30.22 | 27.92 | 17.31 | 3.52 | 12.3 | 86 |
According to the results of the table, the difference of the pH value, the total nitrogen content and the effective phosphorus content of the soil can be obviously seen to reach a significant level, so that the waxberry quality regulator has a certain effect on soil modification. Meanwhile, the application of the waxberry quality regulator promotes the absorption of nutrient substances by the waxberry tree root system, so that the quality of fruits is improved, the waxberry meets the commercial secondary fruit standard, the content of organic acid is low, and the waxberry quality regulator can effectively accelerate the metabolism of the physiological activities of the waxberry and promote the accumulation of sugar, so that the detection finds that the content of vitamin C and soluble sugar of the waxberry is higher. In conclusion, the waxberry quality regulator can enable the waxberry trees at the end of full fruits to continue to bear big fruits with moderate hardness. As the phyllobacterium and the bulimia form dominant bacterial groups in the soil and inhibit the proliferation of bacteria, the number of cracked fruits is obviously reduced, and therefore, the conclusion that the waxberry quality regulator has a certain effect on preventing and treating the novel debilitation of waxberries can be obviously obtained.
In addition, based on the comparison of the non-dense branches and the dense branches, the regulator plays a very excellent role in regulating the development of the dense branches in the Yang Meicheng flowering phase, and the quality of the dense branches is certainly reduced to a certain extent compared with that of the non-dense branches, so that the nutrition value and the single fruit size are reduced, and the regulator still has overall better quality. Therefore, for the plantation with part of high yield requirement, the fruit harvesting amount of common quality can be obviously improved, and of course, the fertilizing amount from the full bloom stage to the full harvest stage should be correspondingly increased when the whole tree grows densely, and further research is needed.
Comparative example 1
Waxberry tree routinely fertilized in the same garden and without debilitation is used as a blank control. If no special description exists, the soil and the fruits are sampled before the same day of harvesting in 7 th month of the current year, and the fruit sampling is only carried out by randomly selecting 8-10 fruits on each myrica rubra tree. And in the full bloom stage, the partial pruning of the dense branches is not carried out, and at least 2 of the fruits are ensured to be dense branch fruits during fruit sampling.
The characterization specifically comprises the following steps:
1. physical and chemical properties of soil:
the pH is measured by an acidometer, the conductivity is measured according to ISO11265:1994 (E) International Standard, the organic matter content is measured by a potassium dichromate-sulfuric acid hydration heat method,the total nitrogen content is determined by Kjeldahl method, the effective nitrogen element content is determined by Kjeldahl method, the effective phosphorus element content is determined by molybdenum-antimony colorimetric method, and the effective potassium element content is determined by NH 4 Ac leaching-flame photometry. Carrying out two times of characterization on physical and chemical indexes of soil, wherein the first time is one day before fertilization, and the second time is sampling before harvesting;
2. the physical index of the waxberry is as follows:
the amino acid content adopts an ninhydrin colorimetric method, the vitamin C content adopts an iodine titration method, the flavonoid content adopts a near infrared hyperspectral method, the phenolic acid content adopts a NaOH neutralization titration method, and the soluble sugar content adopts an anthrone colorimetric method;
3. the quality index of the waxberry fruits is as follows:
measuring transverse diameter and longitudinal diameter of the waxberry fruits by a vernier caliper, measuring single fruit weight of the waxberry fruits by a balance, measuring fruit hardness by a TA-XTplus texture instrument, measuring the content of soluble solids by a handheld glycometer, and recording the average value;
4. good fruit rate:
the method is characterized in that the method is carried out after all the fruits are harvested, the surfaces of the fruits are clean and have no mildew spots, the fruit shape is correct, the scars account for less than 10% of the fruit surface, the fruit rate is calculated, the formula is as follows, the number of the fruits is (N/N) multiplied by 100%, and the number of the fruits is N;
all experimental data were analyzed using GraphPad Prism Version 10.0.2 software and the data are expressed as mean ± standard deviation. One-way analysis of variance (ANOVA) using Tukey test, with at least 3 replicates per treatment, results were as follows * Indicating that the treatment has significant differences compared to the control, i.e. P<0.05）。
The sample characterization results are as follows.
Soil sampling physical and chemical indexes one day before fertilization:
| pH | conductivity (mu s/cm) | Organic matter (g/kg) | Total nitrogen (mg/kg) | Available nitrogen (mg/kg) | Available phosphorus (mg/kg) | Effective potassium (mg/kg) |
| 4.63±0.35 | 96.97±28.11 | 32.30±7.86 | 1.42±0.33 | 120.11±27.22 | 8.13±7.09 | 131.22±10.10 |
Sampling soil sampling physical and chemical indexes before harvesting:
| pH | conductivity (mu s/cm) | Organic matter (g/kg) | Total nitrogen (mg/kg) | Available nitrogen (mg/kg) | Available phosphorus (mg/kg) | Effective potassium (mg/kg) |
| 4.31±0.22 * | 99.96±13.81 | 23.21±0.56 | 1.30±0.11 * | 111.81±3.25 | 31.06±6.17 | 126.08±12.96 |
Non-dense branch red bayberry physiological index:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.997 | 1331.395 | 0.620 | 1.220 | 107.02 |
The quality index of the non-dense branch red bayberry fruits:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 30.71 | 29.92 | 19.01 | 3.60 | 13.1 | 93 |
The physiological index of the dense branch red bayberry:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.611 | 1197.85 | 0.561 | 1.202 | 93.55 |
The quality index of the thick-branch waxberry fruits is as follows:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 23.31 | 20.02 | 13.98 | 3.33 | 10.98 | 66 |
From comparison of the characterization results and the example 1, the regulator can play a very good role in improving the waxberry debilitation when being applied in the full bloom stage, so that the quality of the waxberry debilitation can be close to or even slightly better than that of the waxberry without debilitation, and is particularly unique in the role of the regulator on the pachytic branches. The comparison of the thick-branch waxberries and the non-thick-branch waxberries can obviously find that various qualities of the waxberries are obviously reduced, the reduction is large in amplitude and obvious, the taste is obviously reduced in the aspect of eating, and the sweetness is obviously reduced. And compared with the embodiment 1, the quality of the dense branch red bayberry is not significantly reduced under the action of the regulator in the embodiment 1, so that the regulator can significantly improve the growth disadvantages of the dense branch when being applied in the full bloom period.
Comparative example 2
The waxberry tree which is conventionally fertilized in the same park and has the novel debilitation is used as a control. If no special description exists, the fruits are sampled before the fruits are harvested in the same day in 7 th month of the current year, and the fruit sampling is carried out by randomly selecting 8-10 fruits on each myrica rubra tree. And in the full bloom stage, the partial pruning of the dense branches is not carried out, and at least 2 of the fruits are ensured to be dense branch fruits during fruit sampling.
The characterization specifically comprises the following steps:
1. the physical index of the waxberry is as follows:
the amino acid content adopts an ninhydrin colorimetric method, the vitamin C content adopts an iodine titration method, the flavonoid content adopts a near infrared hyperspectral method, the phenolic acid content adopts a NaOH neutralization titration method, and the soluble sugar content adopts an anthrone colorimetric method;
2. the quality index of the waxberry fruits is as follows:
measuring transverse diameter and longitudinal diameter of the waxberry fruits by a vernier caliper, measuring single fruit weight of the waxberry fruits by a balance, measuring fruit hardness by a TA-XTplus texture instrument, measuring the content of soluble solids by a handheld glycometer, and recording the average value;
3. good fruit rate:
the excellent fruit rate is characterized in that all the fruits are harvested, the surfaces of the fruits are clean and have no mildew spots, the fruit shapes are right, the scars account for less than 10% of the fruit surfaces, the excellent fruit rate is calculated, and the formula is shown as (N/N) multiplied by 100%, wherein N is the number of the excellent fruits, and N is the harvesting number.
All experimental data were analyzed using GraphPad Prism Version 10.0.2 software and the data are expressed as mean ± standard deviation. One-way analysis of variance (ANOVA) using Tukey test, with at least 3 replicates per treatment, results were as follows * Indicating that the treatment has significant differences compared to the control, i.e. P<0.05）。
The sample characterization results are as follows.
Non-dense branch red bayberry physiological index:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.713 | 1189.62 | 0.573 | 1.106 | 101.03 |
The quality index of the non-dense branch red bayberry fruits:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 26.99 | 24.63 | 17.02 | 3.51 | 10.61 | 71 |
The physiological index of the dense branch red bayberry:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.331 | 1126.61 | 0.532 | 1.026 | 91.33 |
The quality index of the thick-branch waxberry fruits is as follows:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 21.02 | 18.67 | 12.73 | 3.36 | 10.32 | 52 |
As is apparent from the comparison results of the above examples 1, 1 and 2, the novel debilitating disease has a great harm to the myrica rubra, the quality of the non-closely-grown myrica rubra fruit of the comparative example 2 is greatly reduced compared with that of the non-closely-grown myrica rubra fruit of the comparative example 1, and the regulator of the present invention can be found to be effective in improving debilitating symptoms and also can be very effective in improving the growth and development conditions of the closely-grown myrica rubra fruit, so that the myrica rubra fruit can be developed more effectively, and although the quality of the myrica rubra fruit is still lower than that of the non-closely-grown myrica rubra fruit, the difference is reduced from the point of view of the results, the microbial agent regulator of the present invention has a positive lifting effect on the nutrient absorption and transportation process of the myrica rubra fruit, and has great value for improving the ordinary myrica rubra yield.
Comparative example 3
The waxberry tree with the novel debilitation is used as a test sample. The Yang Meicheng flowering phase regulator prepared in the example 1 and silkworm excrement organic fertilizer are mixed according to the mass ratio of 0.01:1, and uniformly and annularly applying the mixture in the dosage ratio of 2000 g/myrica rubra in the middle 2 days of 2023. The experimental place is in a red bayberry orchard of Jin Huashi lanxi in Zhejiang province, and near 30 red bayberry trees which are planted in the same period, grow for 8 years and have premature senility symptoms are planted in the red bayberry orchard, and the red bayberry trees belong to Dongkui species, and have nearly 200 red bayberry trees of the same variety in total. And the other period adopts conventional fertilization. If no special description exists, the fruits are sampled before the fruits are collected in the same day in 7 th month of the current year, and the fruit sampling is carried out by randomly selecting 8-10 fruits on each trunk of the myrica rubra. And in the full bloom stage, the partial pruning of the dense branches is not carried out, and at least 2 of the fruits are ensured to be dense branch fruits during fruit sampling.
The characterization specifically comprises the following steps:
1. the physical index of the waxberry is as follows:
the amino acid content adopts an ninhydrin colorimetric method, the vitamin C content adopts an iodine titration method, the flavonoid content adopts a near infrared hyperspectral method, the phenolic acid content adopts a NaOH neutralization titration method, and the soluble sugar content adopts an anthrone colorimetric method;
2. the quality index of the waxberry fruits is as follows:
measuring transverse diameter and longitudinal diameter of the waxberry fruits by a vernier caliper, measuring single fruit weight of the waxberry fruits by a balance, measuring fruit hardness by a TA-XTplus texture instrument, measuring the content of soluble solids by a handheld glycometer, and recording the average value;
3. good fruit rate:
the method is characterized in that the method is carried out after all the fruits are harvested, the surfaces of the fruits are clean and have no mildew spots, the fruit shape is correct, the scars account for less than 10% of the fruit surface, the fruit rate is calculated, the formula is as follows, the number of the fruits is (N/N) multiplied by 100%, and the number of the fruits is N;
all experimental data were analyzed using GraphPad Prism Version 10.0.2 software and the data are expressed as mean ± standard deviation. One-way analysis of variance (ANOVA) using Tukey test, with at least 3 replicates per treatment, results were as follows * Indicating that the treatment has significant differences compared to the control, i.e. P<0.05）。
The sample characterization results are as follows.
Non-dense branch red bayberry physiological index:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 3.117 | 1402.361 | 0.653 | 1.239 | 108.122 |
The quality index of the non-dense branch red bayberry fruits:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 32.62 | 31.01 | 19.02 | 3.61 | 13.2 | 91 |
The physiological index of the dense branch red bayberry:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.516 | 1236.51 | 0.559 | 1.042 | 93.57 |
The quality index of the thick-branch waxberry fruits is as follows:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 24.41 | 20.39 | 13.03 | 3.29 | 10.36 | 56 |
It can also be seen from the above results that the effects described for the different periods of administration are different for the modulators of the present invention. The quality of the non-closely branched waxberry fruits after application at about 20 d a full bloom stage is somewhat better than that of full bloom stage application, but less optimized, and does not produce a significant beneficial improvement for the closely branched fruits. This is mainly because the regulator of the invention is indeed able to enhance nutrient delivery in myrica rubra by improving nutrient absorption and the synergistic effect of fungus cultivation, but the nutrient delivery trend is different in different periods, and when applied in the middle of 2 months, the nutrient can be more significantly preferentially supplied to non-dense branches. Therefore, if the development condition, the fruiting rate and the fruiting quality of the dense branches are to be optimized, the use period is still required to be adjusted to the full bloom period.
Comparative example 4
A Yang Meicheng flowering phase regulator prepared by the method of:
1) Preparing a molasses culture medium, inactivating the molasses culture medium at 121 ℃ for 20min, sub-packaging, and inoculating microbacterium phyllosum and greedy bacteria into one culture medium, wherein the inoculation ratio of the microbacterium phyllosum to the greedy bacteria is 1:1, culturing 48 h by time shaking to obtain fermentation broth, wherein the concentration of bacteria in the obtained fermentation broth is about 9.3X10 9 CFU/mL, filtering and sterilizing to obtain fermentation filtrate;
2) Inoculating rhodopseudomonas palustris as photosynthetic bacteria into the other culture medium, and culturing under sealed condition 48. 48 h until the bacterial concentration is about 8.0X10 9 CFU/mL, and obtaining filtrate through centrifugation, filtration and sterilization;
3) Taking out fermentation filtrate, wherein the volume ratio of the fermentation filtrate to the filtrate is 9: and 1, adding the filtrate in proportion, and uniformly mixing to obtain the Yang Meicheng flowering phase regulator.
The waxberry tree with the novel debilitation is used as a test sample. The Yang Meicheng flowering phase regulator prepared by the method and silkworm excrement organic fertilizer are mixed according to the mass ratio of 0.01:1, and evenly and annularly applying the mixture according to the dosage ratio of 2000 g/myrica rubra tree after the male flowers enter the full bloom stage in the last ten days of 3 months of 2023. The experimental place is in a red bayberry orchard of Jin Huashi lanxi in Zhejiang province, and near 30 red bayberry trees which are planted in the same period, grow for 8 years and have premature senility symptoms are planted in the red bayberry orchard, and the red bayberry trees belong to Dongkui species, and have nearly 200 red bayberry trees of the same variety in total. And the other period adopts conventional fertilization. If no special description exists, the fruits are sampled before the fruits are harvested in the same day in 7 th month of the current year, and the fruit sampling is carried out by randomly selecting 8-10 fruits on each myrica rubra tree. And in the full bloom stage, the partial pruning of the dense branches is not carried out, and at least 2 of the fruits are ensured to be dense branch fruits during fruit sampling.
The characterization specifically comprises the following steps:
1. the physical index of the waxberry is as follows:
the amino acid content adopts an ninhydrin colorimetric method, the vitamin C content adopts an iodine titration method, the flavonoid content adopts a near infrared hyperspectral method, the phenolic acid content adopts a NaOH neutralization titration method, and the soluble sugar content adopts an anthrone colorimetric method;
2. the quality index of the waxberry fruits is as follows:
measuring transverse diameter and longitudinal diameter of the waxberry fruits by a vernier caliper, measuring single fruit weight of the waxberry fruits by a balance, measuring fruit hardness by a TA-XTplus texture instrument, measuring the content of soluble solids by a handheld glycometer, and recording the average value;
3. good fruit rate:
the excellent fruit rate is characterized in that all the fruits are harvested, the surfaces of the fruits are clean and have no mildew spots, the fruit shapes are right, the scars account for less than 10% of the fruit surfaces, the excellent fruit rate is calculated, and the formula is shown as (N/N) multiplied by 100%, wherein N is the number of the excellent fruits, and N is the harvesting number.
All experimental data were analyzed using GraphPad Prism Version 10.0.2 software and the data are expressed as mean ± standard deviation. Singles using Tukey testAnalysis of variance (ANOVA) of factors, at least 3 replicates per treatment, results were as follows * Indicating that the treatment has significant differences compared to the control, i.e. P<0.05）。
The sample characterization results are as follows.
The physiological index of the dense branch red bayberry:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 3.037 | 1459.058 | 0.453 | 1.351 | 87.742 |
The quality index of the thick-branch waxberry fruits is as follows:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 29.33 | 29.67 | 17.31 | 3.20 | 14.1 | 81 |
The phyllobacterium phyllum produces an antibacterial substance with a purine structure in the fermentation process, and the greedy phage can effectively regulate and control the quality and maturity of fruits, and the filtrate of the phyllobacterium phyllum and the greedy phage is used for preparing a regulator, that is, no active strain exists in the regulator, harmful bacteria in soil continue to grow, and the absorption process of tree root systems to nutrients is destroyed. According to the results in the table, the content of soluble sugar in the waxberry fruits is obviously reduced, and according to the results, the accumulation of organic matters in the waxberry can be effectively improved by the combination of She Zhuangwei bacillus and long-acting release after the fermentation of the bulimia, so that the full and sweet big fruits are obtained.
Comparative example 5
A Yang Meicheng flowering phase regulator prepared by the method of:
1) Preparing a molasses culture medium, inactivating the molasses culture medium at 121 ℃ for 20min, sub-packaging, and inoculating microbacterium phyllosum and greedy bacteria into one culture medium, wherein the inoculation ratio of the microbacterium phyllosum to the greedy bacteria is 1:1, culturing 48 h by time shaking to obtain fermentation broth, wherein the concentration of bacteria in the obtained fermentation broth is about 9.3X10 9 CFU/mL；
2) Taking out fermentation filtrate, wherein the volume ratio of the fermentation filtrate to deionized water is 9: and 1, adding deionized water in proportion, and uniformly mixing to obtain the Yang Meicheng flowering phase regulator.
The waxberry tree with the novel debilitation is used as a test sample. The Yang Meicheng flowering phase regulator prepared by the method and silkworm excrement organic fertilizer are mixed according to the mass ratio of 0.01:1, and evenly and annularly applying the mixture according to the dosage ratio of 2000 g/myrica rubra tree after the male flowers enter the full bloom stage in the last ten days of 3 months of 2023. The experimental place is in a red bayberry orchard of Jin Huashi lanxi in Zhejiang province, and near 30 red bayberry trees which are planted in the same period, grow for 8 years and have premature senility symptoms are planted in the red bayberry orchard, and the red bayberry trees belong to Dongkui species, and have nearly 200 red bayberry trees of the same variety in total. And the other period adopts conventional fertilization. If no special description exists, the fruits are sampled before the fruits are harvested in the same day in 7 th month of the current year, and the fruit sampling is carried out by randomly selecting 8-10 fruits on each myrica rubra tree. And in the full bloom stage, the partial pruning of the dense branches is not carried out, and at least 2 of the fruits are ensured to be dense branch fruits during fruit sampling.
The characterization specifically comprises the following steps:
1. the physical index of the waxberry is as follows:
the amino acid content adopts an ninhydrin colorimetric method, the vitamin C content adopts an iodine titration method, the flavonoid content adopts a near infrared hyperspectral method, the phenolic acid content adopts a NaOH neutralization titration method, and the soluble sugar content adopts an anthrone colorimetric method;
2. the quality index of the waxberry fruits is as follows:
measuring transverse diameter and longitudinal diameter of the waxberry fruits by a vernier caliper, measuring single fruit weight of the waxberry fruits by a balance, measuring fruit hardness by a TA-XTplus texture instrument, measuring the content of soluble solids by a handheld glycometer, and recording the average value;
3. good fruit rate:
the excellent fruit rate is characterized in that all the fruits are harvested, the surfaces of the fruits are clean and have no mildew spots, the fruit shapes are right, the scars account for less than 10% of the fruit surfaces, the excellent fruit rate is calculated, and the formula is shown as (N/N) multiplied by 100%, wherein N is the number of the excellent fruits, and N is the harvesting number.
All experimental data were analyzed using GraphPad Prism Version 10.0.2 software and the data are expressed as mean ± standard deviation. Detection using TukeyThe experiments were subjected to one-way analysis of variance (ANOVA) with at least 3 replicates per treatment, with the following results [ (] * Indicating that the treatment has significant differences compared to the control, i.e. P<0.05）。
The sample characterization results are as follows.
The physiological index of the dense branch red bayberry:
| total amino acids (mg/g) | Total vitamin C ([ mu ] g/g) | Total flavonoid (mg/g) | Total phenolic acids (mg/g) | Total soluble sugar (mg/g) |
| 2.819 | 1290.372 | 0.538 | 1.170 | 99.22 |
The quality index of the thick-branch waxberry fruits is as follows:
| transverse diameter (mm) | Longitudinal diameter (mm) | Single fruit weight (g) | Hardness (N) | Soluble solids (%) | Excellent fruit yield (%) |
| 29.33 | 29.12 | 18.25 | 3.27 | 11.8 | 82 |
In the invention, the photosynthetic bacteria rhodopseudomonas palustris metabolized to produce substrates, intermediate reactants, energy, reducing substances and the like, but in the embodiment, the filtrate of the photosynthetic bacteria rhodopseudomonas palustris metabolized is not added, that is, the regulator is not added with metabolic active substances such as cytokinin and auxin, so that the growth speed of She Zhuangwei bacillus and greedy phages is slower, the growth of Frankia actinomycetes in soil is slowed down, the quantity of fungi and fusarium oxysporum is relatively increased, the accumulation of nutrients such as waxberry amino acid, vitamin C and the like is influenced, and the obtained fruits are smaller.
Claims (9)
1. A method for preparing Yang Meicheng flowering phase regulator is characterized in that,
the method comprises the following steps:
1) Preparing a culture medium, sub-packaging after inactivation, inoculating strains into one culture medium, and carrying out timed shaking culture to obtain a fermentation broth;
2) Inoculating strain into another culture medium, sealing, standing for culturing, centrifuging, filtering, and inactivating to obtain filtrate;
3) Taking out the fermentation liquor, adding the filtrate, and uniformly mixing to obtain the Yang Meicheng flowering phase regulator.
2. A method of preparing a Yang Meicheng flowering phase regulator according to claim 1,
the culture medium in the step 1) is a molasses culture medium.
3. A method of preparing a Yang Meicheng flowering phase regulator according to claim 1,
the strains in the step 1) are microbacterium phyllosum and greedy bacteria;
the inoculation ratio of the microbacterium phyllosum to the greedy bacteria is 1: (0.9-1.1).
4. A process for the preparation of a Yang Meicheng flowering phase regulator according to claim 1 or 3,
the concentration of bacteria in the fermentation liquor obtained in the step 1) is 1 multiplied by 10 9 ～1×10 10 CFU/mL。
5. A method of preparing a Yang Meicheng flowering phase regulator according to claim 1,
the strain in the step 2) is photosynthetic bacteria rhodopseudomonas palustris, and the culture is carried out in a sealed static culture process until the concentration of the photosynthetic bacteria rhodopseudomonas palustris is 1 multiplied by 10 9 ～1×10 10 CFU/mL。
6. A method of preparing a Yang Meicheng flowering phase regulator according to claim 1,
the dosage volume ratio of the fermentation liquor to the filtrate in the step 3) is 9: (0.9-1.1).
7. A Yang Meicheng flowering phase regulator obtainable by the process of claims 1 to 6.
8. A Yang Meicheng flowering phase regulator as claimed in claim 7,
when the Yang Meicheng flowering phase regulator is used, the Yang Meicheng flowering phase regulator and a microbial carrier are mixed according to the proportion of 0.008-0.012: mixing the materials according to the mass ratio of 1;
the microbial carrier is an organic fertilizer.
9. A Yang Meicheng flowering phase regulator as claimed in claim 7 or 8,
the Yang Meicheng flowering phase regulator is applied in a loop according to the dosage ratio of 1900-22000 g per myrica rubra tree 1-3 w before Yang Meicheng flowering phase.
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