CN109275665B - Application of all-trans retinoic acid in regulation of locust egg development - Google Patents
Application of all-trans retinoic acid in regulation of locust egg development Download PDFInfo
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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
- A01N49/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group, wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group, wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics thereof
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- 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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/06—Unsaturated carboxylic acids or thio analogues thereof; Derivatives thereof
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
- A61K31/203—Retinoic acids ; Salts thereof
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Abstract
The invention discloses application of all-trans retinoic acid in regulation of locust egg development. The invention treats the locusts migratorius of the mother generation by feeding and injecting all-trans retinoic acid solutions with different concentrations respectively so as to regulate and control the hatching rate of locusts eggs of the next generation. The experimental results prove that: the method has the advantages that the locusts migratoria of the next generation can be remarkably reduced in incubation rate by treating the locusts migratoria of the all-trans type retinoic acid, and the level of the all-trans type retinoic acid of the locusts migratoria of the next generation plays an important role in the development of the locusts of the next generation. The invention provides a new method for developing a novel pesticide to reduce the population quantity of locusts in the next generation after overwintering.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to application of all-trans retinoic acid in regulation of locust egg development.
Background
Locust is a historically important agricultural pest in China, has the characteristics of multiple varieties, great influence and frequent outbreak and disaster formation, and causes great loss to national economy, particularly agricultural production, in China. In recent years, with the increasing trend of global warming, locusts with high density are frequently generated, the risk of long-distance migratory flight is aggravated, the grain production in the agricultural main producing area of China is seriously threatened, the traditional pest control mainly comprises emergency control and chemical control, the pest control excessively depends on chemical pesticides, and the prospective development of novel green pollution-free prevention and control agents is urgently needed.
Diapause is the inherited physiological response of insects that evolves as the insects adapt to the periodic changes in the environment for a long time. The locusts of the mother generation can influence diapause of locusts of the next generation by sensing light and temperature. The research shows that: the development state of the next generation of locust eggs is closely related to the effect of the mother generation, and the mother generation locusts treated by different light can convert light signals into substances such as proteins and secondary metabolites to be transferred to the next generation of locust eggs, so that the difference of hatchability is finally caused. The photoreceptors such as opsin and the like have important effects on the sensitization of the maternal locust, and the all-trans retinoic acid, which is an important substance in the opsin metabolic process, is also very important for transmitting optical signals.
Disclosure of Invention
An object of the present invention is to provide a novel use of all-trans retinoic acid.
The invention provides application of all-trans retinoic acid in any one of a1) -a 7):
a1) regulating and controlling the development of locust eggs;
a2) preparing a product for regulating and controlling the development of locust eggs;
a3) the population quantity of the locusts is reduced;
a4) preparing a product for reducing the population quantity of the locust;
a5) controlling locusts;
a6) preparing a product for preventing and controlling locust;
a7) locust insecticides or locust inhibitors have been developed.
In the above application, the regulation is inhibition; the control of the development of the locust eggs is embodied in the reduction of the hatching rate of the locust eggs.
Another objective of the invention is to provide a method for inhibiting the development of locust eggs.
The method for inhibiting the development of the locust eggs comprises the following steps: improving the level of all-trans retinoic acid in the body of the maternal locust, thereby realizing the inhibition of the development of the next generation egg of the locust.
In the method, the method for improving the level of all-trans retinoic acid in the maternal locust is to treat the locust with the all-trans retinoic acid. Inhibiting the development of the next generation egg of the locust is embodied in inhibiting the hatching rate of the next generation egg of the locust.
Further, the method of treatment may be injection or feeding.
The injection dosage can be 3-10 mug of all-trans retinoic acid per head; preferably 3. mu.g.
The feeding dosage can be 30-100 mu g of all-trans retinoic acid per head; preferably 30. mu.g.
Further, the injection method can be that the all-trans retinoic acid solution is injected from an internode membrane between the second abdominal node and the third abdominal node of the locust abdomen by using a micro-injector. The all-trans retinoic acid solution is obtained by uniformly mixing all-trans retinoic acid and dimethyl sulfoxide solution, and the concentration of the all-trans retinoic acid solution can be specifically 0.6 mg/mL. The number of injections may be 1.
The feeding method can be to feed the locust by adding the all-trans retinoic acid solution into artificial locust feed. The proportion of the all-trans retinoic acid to the artificial locust feed can be specifically 0.15 mg: 1g of the total weight of the composition. The feeding is continuously carried out for 5 days, and the feeding is carried out for 1 time at 8:00 a day in the morning.
It is a final object of the present invention to provide a new use of the above method.
The invention provides application of the method in locust control.
The invention also provides application of the method in reducing the population quantity of the locust.
The invention treats the locusts migratorius of the mother generation by feeding and injecting all-trans retinoic acid solutions with different concentrations respectively so as to regulate and control the hatching rate of locusts eggs of the next generation. The experimental results prove that: the method has the advantages that the locusts migratoria of the next generation can be remarkably reduced in incubation rate by treating the locusts migratoria of the all-trans type retinoic acid, and the level of the all-trans type retinoic acid of the locusts migratoria of the next generation plays an important role in the development of the locusts of the next generation. The invention provides a new method for developing a novel pesticide to reduce the population quantity of locusts in the next generation after overwintering.
Drawings
FIG. 1 shows the hatching rate of locusts eggs laid by migratory locusts after injection and feeding of all-trans retinoic acid. FIG. 1A shows the hatchability of locusts laid by locusts injected with 3. mu.g/head all-trans retinoic acid treatment group and with dimethyl sulfoxide injection control group; FIG. 1B shows the hatchability of locusts laid by locusts injected with 0.03. mu.g/head all-trans retinoic acid treatment group and with dimethyl sulfoxide injection control group; FIG. 1C shows the hatching rate of locust eggs from migratory locusts fed to the treatment group of 30. mu.g/head all-trans retinoic acid and the control group of dimethyl sulfoxide.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
The test insect sources in the following examples: purified population of migratory locusts were continuously fed for 20 generations in the laboratory without any exposure to chemicals. The breeding process is as follows: incubating locusta migratoria locusta eggs in an intelligent artificial climate box under the following incubation conditions: the temperature is 28 +/-1 ℃, the relative humidity is 60%, and the illumination time and the dark time are 10h and 14 h. And (3) transferring locusta nymphs hatched at the same time and with the same size to a pest breeding cage (60cm multiplied by 50cm multiplied by 60cm) and breeding the locusts with fresh wheat seedlings until the locusts become adults. The breeding conditions were as follows: the temperature is 28 +/-1 ℃, the relative humidity is 60%, and the illumination time and the dark time are 10h and 14 h.
In the following examples dimethyl sulfoxide is the product of Solarbio, all-trans retinoic acid is the product of Sigma, oven dried wheat bran is the product of Hei Rui Heng Tong technology Inc. (specification: 2500 g/bag), corn oil is the product of Hei Va technology Inc. (specification: 500 mL/bottle; purity: reagent grade; brand: source leaf).
The main reagents in the following examples: vitamin B, vitamin C, folic acid, oryzanol, glucose and the like are all domestic analytical purities.
The main instruments in the following examples: an ultra-clean bench (Shanghai Bingxu industries, Ltd.), a 1mL micro-injector (Burkard Scientific Co.), a PAX100-3 automatic micro-dropper (Burkard Scientific Co.), an intelligent climatic chamber PRX-450C (Ningbo Saifu laboratory instruments Co., Ltd.), an electric heating constant temperature air-blast drying box DHG-9140A (Shanghai Shuangxu electronics Co., Ltd.), and an autoclave YXQ-LS-50SII (Shanghai Bingmai industries, Ltd.).
Example 1 application of all-trans retinoic acid in inhibition of locust egg development
First, inject all-trans vitamin A acid and process the migratory locust
1. Preparation of all-trans retinoic acid solution
Dissolving all-trans retinoic acid in dimethyl sulfoxide to prepare all-trans retinoic acid solutions with concentrations of 0.6mg/mL and 0.006mg/mL respectively.
2. Selecting locusta migratoria to be treated
And (4) selecting female locusts with consistent development progress (within 48h of emergence) for microinjection.
3. All-trans retinoic acid treated migratory locust
Treating locusta migratoria with all-trans retinoic acid solution by injection, and dividing into the following groups according to different treatment doses:
injection of 3 μ g/head all-trans retinoic acid: injecting 5 μ L of all-trans retinoic acid solution with concentration of 0.6mg/mL into the locust body from the internode membrane between the second and third abdominal nodes of the locust body by using a micro-injector, wherein the needle of the micro-injector is parallel to the body of the locust body, so as to avoid damaging internal organ tissues of the locust body.
Injection of 0.03 μ g/head all-trans retinoic acid: injecting 5 μ L of all-trans retinoic acid solution with concentration of 0.006mg/mL from internode membrane between second and third abdominal nodes of locust abdomen with micro-injector, wherein needle of the injector is parallel to abdomen to avoid injury of internal organ tissue of locust.
Injection of dimethyl sulfoxide: injecting 5 μ L of dimethyl sulfoxide solution into the abdomen of locust through internode membrane between the second and third abdominal nodes with a micro-injector, wherein the needle of the injector is parallel to the abdomen to avoid injury to internal organ tissue of locust.
Each treatment was 5 replicates, each replicate 5 locusts.
4. Raising after treatment
And mixing all the treated female locusts with the same amount of male adult, and culturing in a constant temperature climate box. The breeding conditions comprise light time, dark time 10h 14h, temperature 28 + -1 deg.C, and relative humidity 60%.
Secondly, feeding all-trans retinoic acid to treat migratory locusts
1. Preparation of artificial feed for migratory locust
The formula of the artificial feed for locusta migratoria is as follows: 30g of dried wheat bran, 0.024g of vitamin B, 0.150g of vitamin C, 0.007g of folic acid, 0.015g of oryzanol, 0.300g of glucose, 30mL of sterilized water and 1.5mL of corn oil.
2. Selecting locusta migratoria to be treated
And (4) selecting female locusts with consistent development progress (within 48h of eclosion) for feeding.
3. All-trans retinoic acid treated migratory locust
Migratory locust is treated with all-trans retinoic acid solution in a feeding manner, and the migratory locust is treated with dimethyl sulfoxide as a control.
Feeding 30 mu g/head of all-trans retinoic acid: feeding an artificial feed added with all-trans retinoic acid at 8:00 a.m. (the artificial feed added with all-trans retinoic acid is obtained by adding 0.25mL of 0.6mg/mL of all-trans retinoic acid solution into 1g of artificial feed), adding 5g of fresh wheat at 12:00 a.m., taking out all feeds at 2:00 a.m., and carrying out starvation treatment until the next day. The above operation was repeated for 5 days.
Feeding dimethyl sulfoxide: feeding artificial feed added with dimethyl sulfoxide at 8:00 am (the artificial feed added with dimethyl sulfoxide is obtained by adding 0.25mL of dimethyl sulfoxide solution into 1g of artificial feed), adding 5g of fresh wheat at 12:00 am, taking out all feeds at 2:00 pm, and starving until the next day. The above operation was repeated for 5 days.
Each treatment was 5 replicates, each replicate 5 locusts.
4. Raising after treatment
And mixing all the treated female locusts with the same amount of male adult, and culturing in a constant temperature climate box. The breeding conditions comprise light time, dark time 10h 14h, temperature 28 + -1 deg.C, and relative humidity 60%.
Third, detection of locust egg hatchability
And respectively detecting the hatchability of the next generation locusts after injecting and feeding all-trans retinoic acid. The method comprises the following specific steps: after the migratory locust is treated by the all-trans retinoic acid for one week, the migratory locust starts mating, a female locust starts laying eggs after one week, the migratory locust lays eggs on sandy soil with the thickness of 10 cm of a bottom pad of a culture cage, and the relative humidity of the sandy soil is 20%. The locusts laid down were collected every two days from the start of laying, transferred to a hatching box with vermiculite, at a relative humidity of 20%. These hatching boxes were cultured in another thermostated climate chamber at a hatching temperature of 28. + -. 1 ℃. After about 3-4 weeks, the larvae began to hatch and the number of larvae was recorded (a). Incubation continued for about 20 days until no more larvae were incubated, and incubation continued for another week. Digging out the rest un-hatched locust eggs from the vermiculite, and recording the number of un-hatched eggs (B). The locusts hatching rate is A/(A + B).
As shown in fig. 1, under the short-day conditions (light time: dark time: 10h:14h, temperature 28 ± 1 ℃, relative humidity 60%), the locusts injected with dmso treatment group had a locusts hatching rate of 16.69%, the locusts injected with 0.03 μ g/head all-trans retinoic acid treatment group had a locusts hatching rate of 17.39%, and the locusts injected with 3 μ g/head all-trans retinoic acid treatment group had a locusts hatching rate of only 2.083%. The locusts injected with 3 μ g/head all-trans retinoic acid significantly decreased in hatchability compared to the control group (dmso-injected group), which was 14.61% lower than the control group (P ═ 0.02248) (fig. 1A); locusts spawned by locusts injected with 0.03 μ g/head of all-trans retinoic acid treated group had a hatchability of 17.39%, and there was no significant difference compared with the control group (group injected with dimethyl sulfoxide treated group) (fig. 1B).
Under the condition of short sunshine (the light time: the dark time is 10h:14h, the temperature is 28 +/-1 ℃, and the relative humidity is 60%), the hatchability of locusts laid by locusts fed to the dimethyl sulfoxide treatment group is 16.69%, and the hatchability of locusts laid by locusts fed to the all-trans retinoic acid treatment group at a dose of 30 mu g/head is 2.798%. Compared with the control group (the group fed with dimethyl sulfoxide), the locusts fed with the group fed with 30 mu g/head of all-trans retinoic acid had significantly reduced locusts hatching rate, which was 13.89% lower than that of the control group (P ═ 0.02496) (fig. 1C).
In conclusion, the drop of the hatching rate of the next generation of locust eggs can be caused by injecting 3 mug/head of all-trans retinoic acid into the female locust with the migratory locust or feeding 30 mug/head of all-trans retinoic acid. And the injection of all-trans retinoic acid has dose dependence, the injection of all-trans retinoic acid has effect when high dose (3 mug/head) is injected, and the injection of all-trans retinoic acid does not have function when low dose (0.03 mug/head) is injected.
Claims (4)
1. A method for inhibiting the development of locust eggs comprises the following steps: improving the level of all-trans retinoic acid in the body of the maternal locust, thereby realizing the inhibition of the development of the next generation egg of the locust;
the method for improving the level of all-trans retinoic acid in the maternal locust body comprises the steps of treating the maternal locust with the all-trans retinoic acid;
the treatment method is injection or feeding;
the injection dosage is 3-10 mu g of all-trans retinoic acid/head; the feeding dosage is 30-100 mu g of all-trans retinoic acid per head.
2. The method of claim 1, wherein: the injection dosage is 3 mu g of all-trans retinoic acid/head; the feeding dosage is 30 mu g of all-trans retinoic acid per head.
3. Use of the method according to claim 1 or 2 for controlling locusts.
4. Use of the method of claim 1 or 2 for reducing the population of locusts.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1452958A (en) * | 2003-05-26 | 2003-11-05 | 复旦大学 | Application of all transconfiguration vitamine A acid in preparing medicine for preventing and treating myocardial reconstution |
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| CN1452958A (en) * | 2003-05-26 | 2003-11-05 | 复旦大学 | Application of all transconfiguration vitamine A acid in preparing medicine for preventing and treating myocardial reconstution |
Non-Patent Citations (2)
| Title |
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| Juvenile Hormone-like Effects of Retinoic Acid in Insect Metamorphosis, Embryogenesis and Reproduction;VACLAV NEMEC等;《J. Insect Physiol.》;19931231;第39卷(第12期);第1083-1093页 * |
| Locust retinoid X receptors: 9-Cis-retinoic acid in embryos from a primitive insect;Shaun M. Nowickyj等;《PNAS》;20080715;第105卷(第28期);第9540-9545页 * |
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