CN109953042B - Inhibitor for growth of acacia taiwan based on leaching liquor of castanopsis gracilis litter - Google Patents
Inhibitor for growth of acacia taiwan based on leaching liquor of castanopsis gracilis litter Download PDFInfo
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- CN109953042B CN109953042B CN201910349273.XA CN201910349273A CN109953042B CN 109953042 B CN109953042 B CN 109953042B CN 201910349273 A CN201910349273 A CN 201910349273A CN 109953042 B CN109953042 B CN 109953042B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/02—Germinating apparatus; Determining germination capacity of seeds or the like
- A01C1/025—Testing seeds for determining their viability or germination capacity
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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
Abstract
The invention relates to a preparation method of an inhibitor for growth of acacia taiwan based on extract of litters of the grahami extract, which comprises the steps of washing a grahami extract sample for 3 times by using distilled water, drying, crushing and sieving. Placing 1g of the screened litter into a 50mL centrifuge tube, adding 15mL 70% ethanol, leaching in a 80 deg.C water bath for 2 times, each time for 1h, cooling, placing into a centrifuge, and setting for 8000r min‑1Centrifuge for 5 minutes, filter, combine the two extracts, add water to 50 mL. Placing the mixture on a rotary evaporator, and after the ethanol is evaporated, diluting to 50mL to prepare a stock solution with the volume ratio of 1: 5. Results obtained after applying the inhibitor to taiwan acacia show that 1: the leaching liquor with the concentration of 5 has the most obvious inhibiting effect on acacia confusa species, can maintain good vital signs, is a good seedling inhibitor, does not use chemicals, and has no pollution to the environment.
Description
Technical Field
The invention belongs to the field of plant growth inhibitors, and particularly relates to a preparation method of an inhibitor for acacia confusa growth based on a leaching solution of a castanopsis gracilis litter.
Background
The rare or endangered plants are used as one of the main bodies of species resources, gene resources and ecological environment, and have important significance for maintaining ecological balance. The method is suitable for interplanting tree species of rare or endangered plants, and is beneficial to water and soil conservation of the habitat, soil improvement, pest control, fire hazard reduction and the like. However, improper mixed mode can cause the main tree species to reject the interplanted tree species and cause malignant competition and toxicity. Plant allelopathy widely exists in mixed forests, is an important source for causing the toxicity, and mainly causes the seed germination, seedling growth, flowering and fructification and the like of mixed trees to be influenced by releasing allelopathy substances to the environment so as to influence the population establishment and updating. The litters serve as one of the main sources of allelochemicals, the leaching liquor of the litters has remarkable germination effect on the seeds of the tested plants, and in recent years, the research is concerned with: studies in Yangli show that the water extract of Larix Gmelini leaf has obvious effect of promoting the germination of Juglans mandshurica seeds and the growth of seedlings, and the germination of seeds and the growth of seedlings are in a descending trend along with the decrease of the concentration of the water extract. Studies of old juan and the like show that the influence of the moso bamboo leaching liquor on the relative contents of the height, the ground diameter and chlorophyll of the castanopsis sclerophylla seedling is totally in a high-concentration inhibition mode, and the low-concentration promotion mode has a double-concentration effect. The influence of the litters and leaves on the seed germination and seedling growth of the Korean pine is researched by old establishment and the like, and the result shows that the leaching liquor of the litters in different forest types is reduced along with the concentration, and the inhibition effect is weakened, disappeared or is changed into promotion. Therefore, the method explains the internal association among species from the perspective of allelopathy, and has a good guiding function for explaining plant individuals, the interaction mechanism among species and plant community succession.
Tannin extract (Castanopsis kawakamii) is a peculiar evergreen broad-leaved tree of the middle and subtropical zone, is extremely precious as a wiggle plant of the third era, and is distributed in nothing but the prefecture, taiwan, jiangxi, guangdong, guangxi, and the like. At present, the research on the Grignard tannin extracts mainly focuses on community ecology, forest gap renewal, the research on litters of natural forests and artificial forests and soil carbon reservoir nutrient banks, the research on rhizosphere soil microorganisms of the Grignard tannin extracts and the artificial forests, the biochemical characteristics of the microorganisms and the litters and the stoichiometry of soil, and the research on symbiotic mixed tree allelopathy of the Grignard tannin extracts is rare. Taiwan acacia serves as a common tree species of a mixed forest and is cultivated in tropical and subtropical regions of southwest and southeast provinces. The growth speed is high, the adaptability is wide, the biomass is large, the water storage property and the wet strength are maintained, the interplanting advantages mainly lie in the nitrogen fixation effect of the interplanting method and the rich nitrogen content in litters, according to the determination, 1hm2 h of litters can provide 3500 kg of litters, and the decomposed litters are equivalent to 500kg of urea. And the total nitrogen and the total phosphorus of the soil in the natural protection area of the tannin extract are extremely deficient, so that the germination of the tannin extract seeds and the regeneration of seedlings are restricted. In order to expand the germplasm resources of the Grignard tannin extract, Taiwan missing is an excellent forest mixed tree species which integrates economic and ecological benefits. It is thus clear that it is necessary to investigate whether the litters of the tannin extract natural forest have chemotoxic effects on the acacia confusa. Therefore, the acacia rabdosia is taken as a research object, the influence of different concentrations of the extract of the litters of the tannin extract on the sprouting of the acacia rabdosia seeds is measured and analyzed, the allelopathy of the litters of the tannin extract on the growth of the acacia rabdosia seeds is analyzed, and the scientific basis is provided for interplanting the natural forest of the tannin extract and the acacia rabdosia.
The method is popularized to the interplanting of the artificial forest of the tannin extract from the experience of interplanting the acacia confusa in the tannin extract and then to the interplanting of the acacia confusa in other mature forests. Provides a new idea for improving the excellent condition of the growth of the target forest. The interplanted tree species not only can play a role in fixing nitrogen, but also can not replace the target tree species, and a balanced relationship exists between the tree species and the tree species.
Disclosure of Invention
The invention aims to provide an inhibitor for the growth of acacia confusa based on a grahami extract. In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of an inhibitor for growth of acacia taiwanensis based on a grahami extract leaching solution specifically comprises the following steps:
(1) pretreatment of litters: washing the tannin extract litter sample with distilled water for 3 times, removing soil and humus attached to the surface, drying at 70 ℃, crushing, and sieving with a 40-mesh sieve;
(2) leaching for 1 time: taking the litters pretreated in the step (1), adding 70% v/v ethanol, uniformly mixing, leaching in a water bath kettle at 80 ℃ for 1h, cooling to room temperature, centrifuging at 8000rpm for 5min, and filtering to obtain filtrate;
(3) leaching for 2 times: adding 70% v/v ethanol with the same volume as that in the step (2) into the filter residue, uniformly mixing, leaching in a water bath kettle at 80 ℃ for 1h, cooling to room temperature, centrifuging at 8000rpm for 5min, and filtering to obtain a filtrate;
(3) and (3) rotary steaming: combining the filtrates obtained in the two leaching steps (2) and (3), adding water to a constant volume of 50mL to prepare a stock solution with a volume ratio of 1:5, placing the stock solution in a rotary evaporation evaporator for rotary evaporation until ethanol is completely evaporated, and then, fixing the volume to 50mL to obtain an inhibitor leaching liquor stock solution;
(4) diluting: and (4) diluting the stock solution in the step (3) by adding distilled water according to the volume ratio for 3 times, 7 times, 15 times, 31 times and 63 times, and preparing five leaching solutions with different concentrations of 1:20, 1:40, 1:80, 1:160 and 1: 320.
The Grignard tannin extract withered sample in the step (1) is obtained by mixing the leaves, shells, branches and skins of the Grignard tannin extract withered according to the mass ratio of 1:1:1: 1.
And (3) the mass-to-volume ratio of the pretreated litter in the step (2) to 70% v/v ethanol is 1: 15.
The application of an inhibitor for growth of acacia taiwanensis based on a grahami extract in inhibiting the growth of acacia taiwanensis specifically comprises the following steps:
(1) selecting seeds of acacia confuse with the same size, disinfecting the seeds for 30min by using 0.5wt% of potassium permanganate, washing the seeds for 3 times by using distilled water, uniformly placing the seeds in culture dishes with 3 layers of filter paper, and uniformly placing 100 seeds in each culture dish;
(2) adding 15mL diluted inhibitor leaching solution into each culture dish, culturing in incubator with photoperiod of 25 deg.C, 12 h, dark period of 15 deg.C, 12 h, and light intensity of 50 nmol-1•s-1The relative humidity is 75% -80%; supplementing 4mL of leaching liquor with corresponding concentration every day, and wetting with filter paper;
(3) taking the day of setting germination as day 1, observing and recording data after day 3, and continuously observing for 15 days; recording the number of germinated seeds every day, counting the number of germinated seeds day by taking the embryonic roots exposed out of the seed coats by 2 mm as a standard, and selecting 20 seeds with consistent embryonic root lengths from a culture dish to measure and record the embryonic root lengths when the seeds are in a full germination period; when no seeds germinate for 3 consecutive days, recording the experiment of the end of germination; and when the growth amount of the radicle is basically unchanged, ending the radicle experiment, and finally calculating the germination rate, the germination vigor, the germination index and the radicle growth condition of the seeds.
The invention has the advantages that:
(1) the invention takes the acacia kaki as a research object, analyzes the influence of the extract of the litters of the grazing tannin extract with different concentrations on the sprouting of the acacia kaki seeds by measuring and analyzing the allelopathy of the litters of the grazing tannin extract on the growth of the acacia kaki seeds, and provides scientific basis for interplanting the natural forest of the tannin kaki and the acacia kaki.
(2) The invention is popularized to the interplanting of the artificial forest of the tannin extract from the experience of interplanting the acacia confusa in the tannin extract and then to the interplanting of the acacia confusa in other mature forests, provides a new idea for improving the excellent condition of the growth of the target forest, ensures that the interplanted tree species not only can play an excellent nitrogen fixation role, but also does not replace the target tree species, and has a balance relationship between the tree species and the tree species.
(3) The invention utilizes the extract of the litters of the grist tannin extract to have obvious inhibition effect on the acacia confusa, can maintain good vital signs, is a good seedling inhibitor, does not use chemical medicines and has no pollution to the environment.
Drawings
FIG. 1 shows the effect of various concentrations of litter extracts on the length of Acacia Taiwan radicle.
Detailed Description
(1) The material and the method are as follows:
collecting litters month by month in a fixed sample plot withering frame of a Sanming tannin extract protection area in Fujian of 2016 years for 9 months, dividing the litters into 4 types (leaves, shells, branches and skins), and counting the types of the litters, wherein the litters are mixed into 1 kg in equal mass ratio. The experimental material was acacia confusa seed. 70% (v/v) ethanol, 0.5wt% potassium permanganate, water bath, centrifuge tube, 50mL volumetric flask, rotary evaporator, slide caliper, petri dish and incubator were prepared in the laboratory for culture of Acacia taiwan seeds.
(2) Preparing a leaching solution:
and washing the returned litter sample with distilled water for 3 times to remove soil and humus attached to the surface, drying and crushing at 70 ℃, and sieving by a 40-mesh sieve. Putting 1g of the screened litter into a 50mL centrifuge tube, adding 15mL 70% (v/v) ethanol, leaching for 1h in a water bath kettle at 80 ℃, cooling, putting into a centrifuge, setting the centrifuge at 8000rpm, centrifuging for 5min, and filtering; the above extraction was repeated once more, and the two extracts were combined and water was added to 50mL (3 replicates). Putting the dried powder on a rotary evaporator, evaporating ethanol, diluting to 50mL to obtain a 1:5 stock solution, adding no litters as a control group according to the method, and diluting the stock solution by adding distilled water according to the volume ratio by 3 times, 7 times, 15 times, 31 times and 63 times to obtain five leaching solutions with different concentrations of 1:20, 1:40, 1:80, 1:160 and 1: 320. Storing in a refrigerator at 4 deg.C for use.
(3) Seed germination experiment:
carefully selecting seeds of acacia confusa with the same size, disinfecting the seeds for 30min by using 0.5wt% of potassium permanganate, washing the seeds for 3 times by using distilled water, uniformly placing the seeds in culture dishes with 3 layers of filter paper, uniformly placing 100 seeds in each culture dish, adding prepared leaching liquor with different concentrations into each culture dish, taking the distilled water as a reference, carrying out 7 treatments in total, repeating each treatment for 3 times, and culturing the seeds in an incubator. The culture conditions were: the photoperiod is 25 ℃, 12 h, the dark period is 15 ℃, 12 h, and the light intensity is 50 nmol-1•s-1And the relative humidity is 75% -80%. The day of germination was set to day 1, and observation was started and data was recorded after day 3 for 15 days. Recording the number of the germinated seeds every day, counting the germination number day by taking the embryonic roots exposed out of the seed coats by 2 mm as a standard, and selecting 20 seeds with basically consistent embryonic root lengths from culture dishes with different concentrations to measure and record the embryonic root lengths when the seeds are in the full germination period. When no seeds germinate for 3 consecutive days, recording the experiment of the end of germination; and when the growth amount of the radicle is basically unchanged, ending the radicle experiment, and finally calculating the germination rate, the germination vigor, the germination index and the radicle growth condition of the seeds.
Germination rate (%) = (number of seeds to germinate/total number of seeds to be tested) × 100%;
germination potential (%) = number of seeds normally germinated until the day when average germination number reaches the highest day/total number of test seeds × 100%;
germination index = ∑ (Gt/Dt)
Wherein: dt is the number of germination days, and Gt is the number of seeds and days corresponding to Dt
The research results are shown in table 1, when the germination rate of the seeds of acacia confuse in the distilled water group is 63%, the germination rates of the seeds are respectively 60%, 57%, 60.5%, 63.5%, 66% and 64% when the seeds are treated by leaching solutions with different concentrations of 1:5, 1:20, 1:40, 1:80, 1:160 and 1:320, the germination potential of the seeds is 50% in the distilled water group, the concentration of the leaching solutions is 13.5%, 46%, 46.5%, 52.5%, 54.5% and 51% in sequence from high to low, the germination index of the seeds is 81.32%, the concentration of the leaching solutions is 56.06, 73.77, 75.98, 84.26, 88.61 and 81.46 in sequence from high to low, as shown in fig. 1, the length of the radicle of the seeds is 30.05mm in the distilled water group, and the concentration of the leaching solutions is 19.5mm, 25.54mm, 33.57mm, 33.54mm, 36.09mm and 33.00mm in sequence from high to low.
TABLE 1 Effect of different concentrations of leach liquors on Geranium formosanum seed Germination
In summary, 1: the leaching liquor with the concentration of 5 has the most obvious inhibiting effect on acacia confusa species, can maintain good vital signs, is a good seedling inhibitor, does not use chemicals, and has no pollution to the environment.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (1)
1. The application of an inhibitor for growth of acacia taiwanensis based on a grahami extract in inhibiting the growth of acacia taiwanensis is characterized by specifically comprising the following steps of:
(1) selecting seeds of acacia confuse with the same size, disinfecting the seeds for 30min by using 0.5wt% of potassium permanganate, washing the seeds for 3 times by using distilled water, uniformly placing the seeds in culture dishes with 3 layers of filter paper, and uniformly placing 100 seeds in each culture dish;
(2) adding 15mL diluted inhibitor leaching solution into each culture dish, culturing in incubator with photoperiod of 25 deg.C, 12 h, dark period of 15 deg.C, 12 h, and light intensity of 50 nmol-1•s-1The relative humidity is 75% -80%; supplementing 4mL of leaching liquor with corresponding concentration every day, and wetting with filter paper;
(3) taking the day of setting germination as day 1, observing and recording data after day 3, and continuously observing for 15 days; recording the number of germinated seeds every day, counting the number of germinated seeds day by taking the embryonic roots exposed out of the seed coats by 2 mm as a standard, and selecting 20 seeds with consistent embryonic root lengths from a culture dish to measure and record the embryonic root lengths when the seeds are in a full germination period; when no seeds germinate for 3 consecutive days, recording the experiment of the end of germination; when the growth amount of the radicle is basically unchanged, the radicle experiment is finished, and finally the germination rate, the germination vigor, the germination index and the radicle growth condition of the seeds are calculated;
the preparation method of the inhibitor leaching liquor in the step (2) comprises the following steps:
1) pretreatment of litters: washing the tannin extract litter sample with distilled water for 3 times, removing soil and humus attached to the surface, drying at 70 ℃, crushing, and sieving with a 40-mesh sieve;
the grazing tannin extract withered sample is obtained by mixing the leaves, shells, branches and skins of the grazing tannin extract withered according to the mass ratio of 1:1:1: 1;
2) leaching for 1 time: taking the litters pretreated in the step 1), adding 70% v/v ethanol, uniformly mixing, leaching in a water bath kettle at 80 ℃ for 1h, cooling to room temperature, centrifuging at 8000rpm for 5min, and filtering to obtain filtrate;
the mass volume ratio of the pretreated litter to 70% v/v ethanol is 1g:15mL;
3) leaching for 2 times: adding 70% v/v ethanol with the same volume as that in the step 2) into the filter residue, uniformly mixing, leaching for 1h in a water bath kettle at 80 ℃, cooling to room temperature, centrifuging at 8000rpm for 5min, and filtering to obtain a filtrate;
3) and (3) rotary steaming: mixing the filtrates obtained in the two leaching steps 2) and 3), adding water to a constant volume of 50mL to prepare a stock solution with a volume ratio of 1:5, placing the stock solution in a rotary evaporation evaporator for rotary evaporation until ethanol is completely evaporated, and then, fixing the volume to 50mL to obtain an inhibitor leaching liquor stock solution;
4) diluting: diluting the stock solution obtained in the step 3) by 3 times by adding distilled water according to the volume ratio to prepare a leaching solution with the concentration of 1: 20.
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