CN113045357A - Proanthocyanidin plant nutrient and application thereof - Google Patents

Abstract

The invention discloses a proanthocyanidin plant nutrient and application thereof. The plant nutrient contains effective dose of procyanidine as an active ingredient, is a pine needle extract or a pine bark extract containing procyanidine, and is prepared by extracting pine needles or pine barks with water and/or an organic solvent. The plant nutrient of the invention has obvious effect of promoting plant growth, and is beneficial to improving the health of crops and improving the yield and quality of the crops. The raw materials used by the invention are biomass components derived from natural plants, and the plant nutrient has the characteristics of safety, easy biodegradation, environmental friendliness and the like, and has great application potential and important social significance in agricultural production.

Description

Proanthocyanidin plant nutrient and application thereof

Technical Field

The invention belongs to the technical field of crops, and particularly relates to a proanthocyanidin plant nutrient and application thereof.

Background

Crop nutrition is the material basis of agricultural production, but at present, excessive application of chemical fertilizers causes serious problems of soil structure deterioration, fertility reduction and the like, seriously affects crop health, crop quality and yield, and simultaneously has adverse effects on food safety and environment. The method for increasing the yield of crops and improving the quality of crops without increasing the using amount of the fertilizer is explored, and then the development of novel crop nutrition products which are environment-friendly, remarkable in fertilizer efficiency and low in cost is an important requirement for the development of modern green agriculture. Elements such as N, P, K in the fertilizer are necessary nutrient elements for plant growth and development, and are also the material basis of the fertilizer efficiency of the conventional fertilizer. By using these elements and their photosynthetic products, plants are able to synthesize primary metabolites, such as amino acids, proteins, nucleotides, polysaccharides, etc., necessary for their growth and development through primary metabolism. However, these substances only guarantee the growth and development of plants, but cannot guarantee the survival and development of plants under natural conditions. Therefore, plants also consume a large amount of primary metabolites and their synthetic precursors and energy to synthesize secondary metabolites such as polyphenols, terpenoids, alkaloids, etc. through secondary metabolism to ensure their survival and development.

Procyanidins (proanthocyanidins) are polyphenolic compounds which are unique to and widely existing in higher plants, are formed by linking hydroxyflavanoids through carbon-carbon bonds, and the main structural units of the polyphenolic compounds are flavan-3-ol or flavan-3, 4-diol, such as (+) -catechin and (-) -epicatechin, and exist in the form of monomers, oligomers or polymers in plants. Procyanidins are widely distributed in crops such as vegetables, fruits, beans, cereals and tea. Procyanidins, an inherent secondary metabolite ubiquitous in plants, are a result of the interaction of plants with the environment (biological and non-biological) during long-term evolution, are a self-protective secondary metabolite produced by the interaction of plants with the external environment, and play an important role in the life activities of plants. Procyanidins are mainly present in plant leaves, stems, fruits, seeds, flowers and outer skins, and are usually present in vacuoles of cells, so that the plants can be protected against microbial or animal damage, and the adaptability of the plants to adverse conditions is improved. The content of procyanidine is inversely proportional to the disease probability of picked fruits and vegetables, and the content of procyanidine is closely related to the disease resistance of plants. Research on the biosynthesis route of procyanidine and the analysis of main functional genes shows that for plants, procyanidine is effectively synthesized and transported, has the physiological functions of regulating seed dormancy and germination, maintaining character quality, resisting ultraviolet rays, resisting diseases and insects, scavenging free radicals and the like, plays an important role in protecting the growth of the plants and is closely related to various characters and qualities of crops.

In the process of procyanidine biosynthesis and transport, a series of protein-encoding genes are required to be expressed and enzymatic reactions are involved, and the enormous energy consumption and metabolic burden of plants are also implied. The plant nutrient is added with procyanidine components, provides precursors for the synthesis of plant polyphenols and other substance components, and is helpful for reducing the substance synthesis burden and energy consumption of plants, regulating the initiation and expression of related genes, and promoting the growth of plants.

In addition, the procyanidine is a functional component of daily fruits and vegetables, has the effects of protecting cardiovascular system, preventing hypertension, resisting tumors, beautifying, improving human microcirculation and the like, and is an internationally recognized natural antioxidant for removing free radicals in a human body. The antioxidant effect of the oligomeric proanthocyanidins in vivo is much higher than that of vitamin E and vitamin C, and the antioxidant capacity in vivo is about 50 times of that of the vitamin E and 20 times of that of the vitamin C. At present, oligomeric proanthocyanidins are widely used in the fields of cosmetics and health foods. Therefore, the plant nutrient based on the procyanidine has the characteristics of safety, easiness in biodegradation, environmental friendliness and the like. However, no documents and patents report about plant nutrients containing proanthocyanidins as active functional ingredients.

Disclosure of Invention

The invention aims to provide a plant nutrient taking procyanidine as an effective component and application thereof.

A nutrition supply mode and a nutrition supply way of the inherent secondary metabolic pathway of the targeted plant life activity provide new strategies and ideas for promoting the growth of crops, improving the quality of characters and developing crop nutrition products. The search for plant endogenous substances capable of assisting the synthesis of plant secondary metabolites is an effective way for developing novel crop nutrition products.

The plant nutrient taking procyanidine as a functional component provides a precursor for the metabolism of substances of crops, reduces the metabolic burden and energy consumption, is beneficial to effectively improving the health condition of the crops, improves the resistance of the crops to biotic and abiotic stress, and promotes the yield increase and the character quality improvement of the crops; meanwhile, the procyanidine is used as a biomass component of a natural plant source, is safe to organisms, is environment-friendly and has low cost. Therefore, the development of the plant nutrient based on the procyanidine promotes the research and development of crop science, and has huge application potential and important social significance in agricultural production.

The first object of the present invention is to provide a plant nutrient characterized by containing an effective amount of procyanidin as an active ingredient.

Specifically, provided is a plant nutrient which can be stored in a stable state and dispersed in a medium for providing nutrition to plants such as soil or an aqueous solution, and which is effectively utilized by plants, based on procyanidins as an active functional ingredient.

Procyanidins used in the present invention are composed of a group of compounds collectively called flavonoids, and mainly refer to a group of compounds containing a condensation product having flavan-3-ol and flavan-3, 4-diol as a structural unit. Therefore, in addition to the above, the proanthocyanidins of the present invention include, for example, flavones, flavonols, flavanonols, isoflavones, catechins (for example, epicatechin gallate, epigallocatechin gallate, etc.), etc., and the proanthocyanidins of the present invention may be flavones, flavonols, flavanonols, isoflavones, and catechins, and include those containing the above, and are all labeled as proanthocyanidins in the present invention.

The procyanidine used in the invention is rich in bark of pine, oak, wild peach and the like, pine needle of pine, fruit and seed of grape, strawberry, cowberry and the like, shell of fruit of lychee, longan, walnut, cocoa bean and the like, seed coat of peanut and the like. The procyanidins contained in these plants are mostly those based on flavan-3-ol and flavan-3, 4-diol as the structural units. The procyanidin can also be processed with various forms of extracts of the above plants or additives, and distributed in market.

In the present invention, proanthocyanidins are preferably a pulverized product or an extract thereof of the above-mentioned proanthocyanidins-rich bark, pine needle, fruit or seed.

Particularly, the extracts of pine needles and pine barks are preferably used, and the extract is preferably used as a raw material of procyanidin.

Preferably, the plant nutrient is pine needle extract or pine bark extract containing procyanidin, and is prepared by extracting pine needles or pine bark with water and/or organic solvent.

Preferably, the pine is masson pine, Chinese pine, slash pine, larch, black pine, red pine, French maritime pine or American longleaf pine.

Preferably, the pine needle extract is prepared by the following method:

taking fresh pine needles, crushing, adding water with the mass 10 times that of the pine needle raw material, soaking for 36 hours at 70 ℃, and filtering to obtain a first leaching solution; adding water which is 5 times of the mass of the pine needle raw material into filter residue, soaking for 24 hours at 70 ℃, and filtering to obtain a second leaching solution; adding water with the mass 3 times of that of the pine needle raw material into filter residues, soaking for 12 hours at 70 ℃, filtering, combining leaching liquor obtained in the three times, and concentrating to obtain the pine needle extract.

Preferably, the pine needle extract is prepared by the following method:

taking fresh pine needles, crushing, adding 40% ethanol aqueous solution with volume fraction 10 times of the mass of the pine needle raw material, soaking for 48 hours at room temperature, and filtering to obtain a first leaching solution; adding 40% ethanol aqueous solution with volume fraction 5 times of the mass of the pine needle raw material into filter residue, soaking for 48 hours at room temperature, and filtering to obtain a second leaching solution; adding 40% ethanol aqueous solution with volume fraction 3 times of the mass of the pine needle raw material into filter residue, soaking for 24 hours at room temperature, filtering, combining three leaching solutions, and concentrating to obtain the pine needle extract.

More preferably, the pine is masson pine.

Preferably, the pine bark extract is prepared by the following method:

taking pine endothelium, crushing, adding 50% ethanol water solution with volume fraction of 8 times of the pine endothelium raw material, soaking for 48 hours, and filtering; adding ethanol water solution with volume fraction of 50% and 3 times of the weight of the pine bark raw material into filter residue, soaking for 24 hours, and filtering; adding ethanol water solution with volume fraction of 50% 3 times of the weight of the pine bark raw material into filter residue, soaking for 24 hours, filtering, combining three leaching solutions, concentrating under reduced pressure, adding water, standing for precipitation, removing supernatant, concentrating under reduced pressure, adding acetone with volume of 4.5 times of the volume of the mixture under stirring, standing until the precipitation is complete, concentrating the supernatant under reduced pressure, and drying to obtain the pine bark extract.

More preferably, the pine is Chinese pine.

The folium Pini extract is preferably prepared from folium Pini belonging to Pinus of Pinus massoniana, Pinus tabulaeformis, slash pine, Larix gmelini, Pinus thunbergii, Pinus densiflora, Pinus maritima, and Pinus palustris. The folium Pini extract of Pinus massoniana (Pinus massoniana Lamb.) is preferred.

The folium Pini extract is obtained by extracting folium Pini with water and/or organic solvent. When water is used, warm water or hot water is used. The organic solvent used for extraction uses organic solvents permitted to be used for manufacturing foods or medicines, for example, methanol, ethanol, propylene glycol, acetone, ethyl acetate, and the like. These water and organic solvent may be used alone or in combination. Particular preference is given to using hot water, aqueous ethanol and aqueous propylene glycol. The method for extracting procyanidin from pine needles is not particularly limited, and for example, a heating extraction method, a supercritical fluid extraction method, or the like may be used. A combination of several approaches may also be employed.

The second purpose of the invention is to provide the application of procyanidine in promoting plant growth and the application of the plant nutrient in promoting plant growth.

The plant nutrient has multiple functions and broad spectrum, is suitable for various agricultural, forestry and animal husbandry crops, such as field crops, vegetables, fruit trees, tea trees and the like, has relatively comprehensive growth promotion effect, is not limited by geographical environment and crop variety factors, and has wide application range. The plant nutrient can effectively improve the yield and quality of crops, has a certain prevention and control function on various plant diseases and insect pests, and can also enhance the capability of the crops for resisting natural disasters.

The plant nutrient can be prepared into any one dosage form suitable for agriculture by using a conventional method in the field, and the better dosage form is water aqua, soluble powder, water-soluble granules or water dispersible granules. For example, the plant nutrient of the present invention may be added to a medium for plant cultivation. In addition, the plant cultivation medium refers to soil or culture medium for planting and cultivating plants, or water manure and water for farmland cultivation irrigation, and the plant nutrient of the invention can be scattered in soil, contained in the culture medium, or added in water manure, water culture and irrigation water. Further, the plant seed may be immersed in the plant nutrient of the present invention or a liquid containing the plant nutrient of the present invention, or the root seedling of the plant or a part of the root or seedling may be immersed in the liquid. The form of use of the plant nutrient of the present invention is not limited thereto. The application amount of the fertilizer is adjusted according to the change of weather conditions or the growth state of crops.

The plant nutrient of the present invention can promote the growth of the overground part of the plant, the underground part of the root, etc., and is effective under low concentration, and has the advantage of no concentration harm. The plant nutrient disclosed by the invention has the advantages of no pollution to the environment, easiness in degradation, high safety and no harm to people, livestock and poultry, and can be used for common dry or paddy fields, interplanting, facility cultivation, hydroponic cultivation, crop rotation farmlands and the like.

The invention at least comprises the following beneficial effects:

1. the plant nutrient has the obvious effect of promoting the growth of plants, and is beneficial to improving the health of crops and improving the yield and the quality of the crops;

2. the plant nutrient has the advantages of easily available raw materials, wide sources, low cost and simple preparation process;

3. the plant nutrient is a natural biomass component of plants, so that the problem of pesticide residue is solved;

4. the plant nutrient of the invention does not belong to phytohormone, and compared with the single use of phytohormone, the risk of phytotoxicity is greatly reduced.

Drawings

FIG. 1 is the product color after plant nutrient reaction (1-procyanidin standard, 2-plant nutrient, 3-blank control);

FIG. 2 is a UV-VIS absorption spectrum of a plant nutrient before (left panel) and after (right panel) the reaction;

FIG. 3 is LC-UV-MS (UV detection wavelength: 280nm) before and after degradation of benzyl mercaptan in pine bark procyanidin extract;

FIG. 4 shows LC-MS molecular ion peaks of benzyl mercaptan degradation products of procyanidin extracted from cortex Pini;

FIG. 5 is the effect of different concentrations of pine needle plant nutrient on the overground part of Arabidopsis plants.

Detailed Description

It should be noted that the specific embodiments described herein are merely illustrative of the invention. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. Other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principles of the invention are intended to be included within the scope of the invention.

Example 1 preparation of proanthocyanidin plant nutrient from pine needles

(1) Taking fresh pinus massoniana needle, and crushing for later use. Adding tap water with the mass 10 times of that of the pine needle raw material, soaking for 36 hours at 70 ℃, and discharging a first leaching solution after filtering; adding water with the mass 5 times of that of the pine needle raw material into filter residues, soaking for 24 hours at 70 ℃, and discharging a second leaching solution after filtering; adding water with the mass 3 times of that of the pine needle raw material into filter residues, soaking for 12 hours at 70 ℃, filtering, combining leaching liquor for three times, and concentrating to prepare a water agent, wherein the mass fraction of the active component procyanidin is 0.01%; thus preparing the pine needle plant nutrient.

(2) Or taking fresh pinus massoniana needle, crushing, adding 40% ethanol water solution with volume fraction 10 times of the mass of the raw material of the pinus massoniana needle, soaking for 48 hours at room temperature, and filtering to obtain a first leaching solution; adding 40% ethanol aqueous solution with volume fraction 5 times of the mass of the pine needle raw material into filter residue, soaking for 48 hours at room temperature, and filtering to obtain a second leaching solution; adding 40% ethanol aqueous solution with volume fraction 3 times of the mass of the pine needle raw material into filter residue, soaking at room temperature for 24 hours, filtering and combining three times of leaching liquor, and concentrating to obtain water agent with the mass fraction of active component procyanidin of 0.01%; thus preparing the pine needle plant nutrient. The pine needle plant nutrient prepared by the method (2) in the subsequent examples is tested.

Example 2 detection of procyanidins in plant nutrients

Taking pine needle plant nutrient as an example. The 4,8 connecting bonds between procyanidine structural units are easy to crack under the action of acid, catechin or epicatechin is formed in the lower unit, and the upper unit becomes a carbocation and loses protons to form yellow-3-alkene-3-alcohol, so that anthocyanin is further formed. Based on the principle, an ultraviolet-visible spectrophotometer method can be adopted to determine the absorbance of the generated product so as to carry out the qualitative and quantitative detection of the procyanidine. The pine needle plant nutrient in example 1 is prepared into a methanol solution with the concentration of 10mg/mL by adopting an n-butanol/hydrochloric acid method, 1mL of the methanol solution is added into a 15mL pressure-resistant glass reaction tube, and then 0.2mL of ferric ammonium sulfate solution (the mass fraction is 2%) and 3.8mL of n-butanol/hydrochloric acid solution (the volume ratio is 95:5) are sequentially added. The reaction tube was closed, the reaction solution was shaken up, reacted at 90 ℃ for 40 minutes, rapidly cooled after the reaction was completed, and the absorbance value at 530nm was measured. Methanol is used as a blank control to replace pine needle plant nutrient. FIG. 1 shows the color of the product of the reaction of pine needle nutrient and procyanidin standard (procyanidin dimer B2, CAS: 29106-49-8), which is characteristically red; FIG. 2 is the UV-VIS absorption spectrum of the pine needle nutrient before and after the reaction, from which it can be found that the reaction product shows characteristic absorption at 530 nm. In the embodiment of the invention, the content of procyanidine is detected according to the method.

Example 3 preparation of proanthocyanidin plant nutrient from pine bark

Taking dried bark of slash pine, and crushing for later use. Adding 70% edible alcohol with volume fraction 8 times of the raw materials, soaking at normal temperature for 48 hours, filtering, and discharging the first leaching solution; adding 70% edible alcohol with the volume fraction of 4 times of the mass, soaking for 24 hours at normal temperature, combining the leaching liquor of the two times, and concentrating to obtain the total extracting solution, wherein the mass fraction of the active ingredients is 30%; pine bark phytonutrients were thus prepared for testing in the subsequent examples.

Example 4 refining of pine bark procyanidin plant Nutrition Agents

Referring to patent (Chinese patent: CN99116183.1), procyanidin refining method comprises collecting dried endothelium of Pinus Marina 100 g, soaking in 800 ml 50% ethanol water solution for 48 hr, filtering, soaking the residue in 300 ml 50% ethanol water solution for 24 hr, and filtering; soaking the residue in 300 ml of 50% ethanol water solution with volume fraction for 24 hours; combining the three extracting solutions, concentrating under reduced pressure to 50 ml, adding water to 200 ml, standing for 12 hours until the precipitation is complete, pouring out the supernatant, concentrating under reduced pressure to about 20 g, adding acetone with 4.5 times volume of the concentrated solution under stirring, standing for 24 hours until the precipitation is complete, concentrating under reduced pressure and drying (vacuum drying, spray drying or freeze drying) the supernatant to obtain light brown powder. The yield is 14 percent, and the content of procyanidin is 68 percent.

Example 5 comparison of procyanidin content in Pinus tabulaeformis and Pinus massoniana needle plant Nutrition

According to the method (2) in the example 1, 250g of each of the pine needles of the Chinese pine and the Chinese red pine is taken to extract procyanidine. And the procyanidin content in the extract was determined according to the procyanidin content determination method in example 2. And (3) measuring results: the content of procyanidine in the pine needle extracts (dry matter) of the Chinese pine and the masson pine needles is respectively 36 percent and 27 percent (mass fraction).

Example 6 molecular Structure and degree of polymerization of procyanidins in pine bark plant Nutrition Agents

The molecular structure and polymerization degree of procyanidine in the pine bark plant nutrient are determined by a hydrochloric acid-benzyl mercaptan degradation method. Solution A: the pine bark plant nutrient prepared in the embodiment 3 is prepared by 95 percent ethanol, and the concentration of the pine bark plant nutrient is 10 mg/mL; solution B: benzyl mercaptan/ethanol (5: 95 by volume) and HCl concentration adjusted to 0.4M with concentrated HCl. Adding 200 mu L of the solution A and 200 mu L of the solution B into the reaction tube, reacting for 30 minutes at 40 ℃, and immediately placing the mixture into a refrigerator at-20 ℃ for storage for later use. When LC-MS analysis is carried out, the reaction solution is injected after passing through a 0.45 mu m microporous filter membrane. LC-MS analysis conditions: the mobile phase A is water (containing 0.1% formic acid, volume ratio), the mobile phase B is methanol, and the elution gradient is 0-10 min, 10-100% B; 10-11 min, 100% B; 11-15 min, 10% B. The analysis results are shown in fig. 3 and 4, and the average degree of polymerization of procyanidin is 16, the terminal unit is mainly catechin, and the chain extension unit is epicatechin according to the retention time, the molecular weight and the chromatographic peak integral area and compared with the standard.

Example 7 Effect of plant Nutrients on growth of the model plant Arabidopsis thaliana

And (3) experimental design: the arabidopsis seedlings are transplanted for 5-7 days. 3 plants per pot (phi 7cm), 5 pots of each plant are treated, the pine needle plant nutrient in example 1 (wherein the mass fraction of the active component procyanidin is 0.01%) is diluted by 100 times and 800 times after one week, the pine needle plant nutrient with corresponding concentration is treated for the second time after two weeks, and the application amount of each plant is 1 ml. The basic culture solution is Hoagland solution.

And (3) test results: the dry weight of the plants was measured 4 weeks after treatment and resulted in an increase of 33.2% and 46.0% in the dry weight of arabidopsis plants compared to the placebo group when applying 100 and 800 times diluted pine needle plant nutrient, respectively (fig. 5).

Example 8 Proanthocyanidins promoting Germination of vegetable seeds

And (3) experimental design: (1) selecting full lettuce seeds, sterilizing with 75% alcohol for 5min, washing with pure water for 3 times, and soaking in pure water for 12 hr; (2) the concentrations of the refined pine bark procyanidin samples (prepared in example 4) prepared with water (containing 1% methanol by volume) were 50, 100, 200, 400 and 800 μ g/mL, respectively; (3) 5 treatments were set: blank control (pure water containing 1% methanol by volume), and above 5 procyanidin concentration gradients, each treatment was repeated 3 times; (4) 50 seeds are inoculated in each culture dish, two layers of filter paper are paved below the culture dish, and 2mL of procyanidine samples are respectively added; (5) placing into a light incubator, culturing at 28 ℃ under the whole day of light condition, and observing and counting the germination number of seeds every 12 hours.

And (3) test results: 50. at concentrations of 100, 200, 400 and 800. mu.g/mL, germination rates were increased by 5%, 13%, 21%, 16% and 7%, respectively, relative to the control group.

Example 9 field fertilizer efficiency test of plant nutrient on eggplant and corn

And (3) experimental design: the experimental object is planted by using a planting tank together with peat soil, and the pine needle plant nutrient prepared by the method of the embodiment 1 (wherein the mass fraction of the active component procyanidin is 0.01%) is sprayed on a treatment group under the condition that other water and fertilizer management conditions are consistent, and the spraying and fertilizing amount is 15L/667m²(ii) a The control group was supplemented with water instead of nutrient solution. Spraying every 15 days for 1 time.

And (3) test results: (1) eggplant: the quality analysis of 100 eggplants in the plant nutrient treatment group and the control group is carried out at random, and the result shows that the average quality of a single eggplant is increased by 9 percent under the plant nutrient addition treatment and reaches the statistical significance level. Eggplant quality detection shows that the application of the plant nutrient can effectively improve the total ash content (increased by 50 percent), protein (increased by 14.5 percent) and carbohydrate (increased by 9 percent) of eggplants. (2) Corn: the plant nutrient is randomly taken to treat 50 corn plants in the control group and the height and the diameter of the ground stem of the corn are measured at 15 days and 30 days after the treatment, and the result shows that the application of the plant nutrient can improve the average growth rate of the corn (increase by 6%) and obviously promote the expansion of the ground stem (increase by 51.5%) in the early growth stage (within 15 days) of the corn. The average corn grain weight is increased by about 3% under the condition of adding the plant nutrient agent by comparing and analyzing the average corn grain weight, and the lifting effect reaches the statistical test level (P < 0.05).

Example 10 influence of pine needle plant nutrient on growth and quality of spring tea

And (3) examining the application effect of the pine needle plant nutrient in the tea garden, and selecting the pine needle plant nutrient as an organic fertilizer by taking no fertilizer as a reference.

The tested tea tree variety: bird seeds (age of 16 years, pH 5.86, organic matter 8.2g/kg, consistent field management level, and severe pruning before sealing the tea garden).

And (3) experimental design: each treatment was repeated 3 times, randomized block arrangement, 54m per cell area²(each row of tea plant has the length of 15m and the width of 1.2 m, each three rows of the tea plant have one cell, and 2 rows are reserved among the cells to be used as protection rows). Applying fertilizer by root irrigation.

Test time, site, fertilization times and dosage: fertilizing at tea base of Hengwei agriculture science and technology Limited in Guizhou by applying fertilizer (containing 0.01% procyanidin) for 2 times in 11/14 th in 2019 and 3/19 th in 2020, with the amount of fertilizer applied being 20L/667m each time²。

The field observation is divided into: (1) observing the tea tree in the spring bud germination period (randomly observing the tea tree germination condition of each cell in 2 months and 27 days in 2020, and observing 3 points in each cell); (2) bud density (random investigation is carried out in 24 days after 3 months, 3 points are searched in each cell, and the area of each point is 0.1m²Calculating the number of new buds); (3) one bud and one leaf with one bud and one hundred bud weight (100 fresh leaves picked from each plot in 3 months and 24 days are randomly selected for weighing); (4) sprout growth potential (sprout growth potential survey is carried out every 5 days from 3 months 10 days to 4 months 20 days).

And (3) test results: the tea trees are fertilized twice in winter and spring, the tea trees can sprout 5 days earlier, the growth rate of young sprout days is improved by 44.7%, the density of sprout heads is increased by 13.92%, the number of one sprout and one leaf is increased by 36.34%, the content of water extract is improved by 4.11%, the content of tea polyphenol is increased by 11.38%, and the total content of catechin is increased by 15.83% (wherein, the content of EGCG is increased by 13.67%, the content of ECG is increased by 12.89%, the content of EGC is increased by 35.44%, the content of catechin is increased by 50%, and the content of epicatechin is increased by 24%).

And (4) test conclusion: the pine needle plant nutrient organic fertilizer can be used for advancing the spring bud germination time of tea trees, and improving the growth rate and the germination density of new tea tree shoots and one-bud and one-hundred-bud seeds. In addition, the pine needle plant nutrient can effectively improve the quality of spring tea, and the water extract, the tea polyphenol content and the total catechin content are obviously improved compared with the fertilizer group.

Example 11 Effect of pine needle plant Nutrition growth and quality of Capsicum annuum

The tested pepper varieties: the variety of the self-bred capsicum annuum of the agricultural institute of Guizhou province is Lai-jian 101 (pod pepper) and Lai-jian 201 (line pepper).

Test site: guiyang and Zunyi test bases of the Guizhou province capsicum research institute; shiqian, Puding, Suiyang 3 demonstration counties.

Test time: 3 months in 2020-10 months in 2020.

And (3) experimental design: the experiment was performed in a randomized block arrangement, with three replicates of each treatment. The pepper is colonized in double rows, and the distance between the strains and the rows is 40 × 120 cm. Dividing into 3 treatments, treating A as pine needle plant nutrient, treating B as Wollo water soluble fertilizer (product type: 12-6-40+ TE; product type: Shanxi Wollo Guo Fengfeng fertilizer Co., Ltd.), and total nutrient (N + P)₂O₅+K₂O) is more than or equal to 60 percent), and the control treatment is clear water. Spraying leaf surface at seedling stage, flowering stage and fruit stage, wherein spraying the leaves with 10L folium Pini plant nutrient (containing active component procyanidin 0.2 wt%) diluted 25 times per mu after A treatment; b, treating the Wolfruit water-soluble fertilizer, diluting by 1000 times and spraying to leaf surfaces, wherein the concentration is 3 kg/mu; control treatments were foliar spray with equal volume of clear water.

And (3) test results:

(1) compared with a blank control, the application of the pine needle plant nutrient increases the plant height of the pepper by 3.79-29.76% and the stem thickness by 5.01-51.22%, wherein the liner pepper variety is the most obvious, and the increases are 16.27-22.94% (plant height) and 27.25-51.22% (stem thickness) respectively.

(2) Compared with a blank control, the pine needle plant nutrient is applied to increase the yield of the linear peppers by 17.64-18.94% and increase the yield of the pod peppers by 4.42-35.25%. In the Zunyi official base test, the application of the pine needle plant nutrient increases the average fruit length of the pepper by 45.11 percent and the average fruit stem by 31.03 percent, and the test results of other tests are similar. The content of phosphorus, potassium, Vc and dry matter in the pepper fruits is respectively increased by 10.01-77.77%, 10.74-82.79%, 9.50-29.08% and 3.76-43.43%. But has no obvious influence on the nitrogen content of the pepper fruits.

The comprehensive experimental results of Guiyang, Zunyi, Shiqian, Puding and Suiyang show that the pine needle plant nutrient has obvious effects on promoting the growth, yield and quality of the pepper.

Claims (10)

1. A plant nutrient comprising an effective amount of proanthocyanidin as an active ingredient.

2. The plant nutrient of claim 1, wherein the pine needle extract or pine bark extract containing procyanidins is prepared by extracting pine needles or pine bark with water and/or an organic solvent.

3. The phytonutrient according to claim 2, wherein the pine is Pinus massoniana, Pinus tabulaeformis, Pinus elliottii, Larix gmelini, Pinus thunbergii, Pinus densiflora, Pinus maritima, or Pinus palustris.

4. The plant nutrient as claimed in claim 2, wherein the pine needle extract is prepared by the following method:

taking fresh pine needles, crushing, adding water with the mass 10 times that of the pine needle raw material, soaking for 36 hours at 70 ℃, and filtering to obtain a first leaching solution; adding water which is 5 times of the mass of the pine needle raw material into filter residue, soaking for 24 hours at 70 ℃, and filtering to obtain a second leaching solution; adding water with the mass 3 times of that of the pine needle raw material into filter residues, soaking for 12 hours at 70 ℃, filtering, combining leaching liquor obtained in the three times, and concentrating to obtain the pine needle extract.

5. The plant nutrient as claimed in claim 2, wherein the pine needle extract is prepared by the following method:

taking fresh pine needles, crushing, adding 40% ethanol aqueous solution with volume fraction 10 times of the mass of the pine needle raw material, soaking for 48 hours at room temperature, and filtering to obtain a first leaching solution; adding 40% ethanol aqueous solution with volume fraction 5 times of the mass of the pine needle raw material into filter residue, soaking for 48 hours at room temperature, and filtering to obtain a second leaching solution; adding 40% ethanol aqueous solution with volume fraction 3 times of the mass of the pine needle raw material into filter residue, soaking for 24 hours at room temperature, filtering, combining three leaching solutions, and concentrating to obtain the pine needle extract.

6. The plant nutrient according to claim 4 or 5, wherein the pine is masson pine.

7. The phytonutrient according to claim 2, wherein said pine bark extract is prepared by the following method:

taking pine endothelium, crushing, adding 50% ethanol water solution with volume fraction of 8 times of the pine endothelium raw material, soaking for 48 hours, and filtering; adding ethanol water solution with volume fraction of 50% and 3 times of the weight of the pine bark raw material into filter residue, soaking for 24 hours, and filtering; adding ethanol water solution with volume fraction of 50% 3 times of the weight of the pine bark raw material into filter residue, soaking for 24 hours, filtering, combining three leaching solutions, concentrating under reduced pressure, adding water, standing for precipitation, removing supernatant, concentrating under reduced pressure, adding acetone with volume of 4.5 times of the volume of the mixture under stirring, standing until the precipitation is complete, concentrating the supernatant under reduced pressure, and drying to obtain the pine bark extract.

8. The plant nutrient of claim 7, wherein the pine tree is Pinus tabulaeformis.

9. Application of procyanidin in promoting plant growth is provided.

10. Use of the plant nutrient of any one of claims 1-8 for promoting plant growth.

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