A preservation method for an experimental population of Psylla pyrisuga

Abstract

The present invention discloses a preservation method for an experimental population of Psylla pyrisuga. In this method, Psylla pyrisuga eggs and 1st instar nymphs are collected and reared with fresh pear tree leaves in a climate chamber until winter-type adults are obtained. A certain density of winter-type adults, pear tree branches that have been dried after removing leaves and branches, and nano-cotton dipped in nutrient solution are placed together in a bottle and preserved at 2°C in the dark. During the preservation period, the nano-cotton dipped in nutrient solution is replaced every 15 days. After covering the bottle mouth with a mesh and opening it, it is irradiated under an ultraviolet lamp at 20°C for 10 minutes, then placed at 20°C under natural light for 4 hours and then preserved again. The preservation method for the experimental population of Psylla pyrisuga of the present invention is highly efficient and easy to operate, can effectively avoid the cumbersome work of long-term retention of the Psylla pyrisuga population, and can also provide Psylla pyrisuga at any time for propagation.

Description

Technical Field

[0001] The present invention relates to a preservation method for an experimental population of Psylla pyrisuga, belonging to the technical field of insect preservation. Background Art

[0002] In the field of orchard pest control technology, Psylla pyrisuga is a seriously harmful pest in the orchard, which is widely distributed in major pear tree production areas in China. Psylla pyrisuga mainly damages with nymphs, often concentrating on the leaves, sucking and feeding along both sides of the main vein on the back, causing the leaves to bend upward, and the new shoots will also be damaged; moreover, it often secretes mucus during the damage process, forming sooty mold, affecting plant photosynthesis, seriously causing the leaves to wither and fall off, reducing the fruit yield and quality. In order to carry out more effective control of Psylla pyrisuga, a large number of experimental populations of Psylla pyrisuga are needed for research work. However, it is a specialized pear tree parasitic pest, and a large number of cultivated pear tree seedlings are required for the laboratory to breed its population, which is very inconvenient. In view of this, it is particularly important to invent a method for long-term preservation of the Psylla pyrisuga population. Based on the above problems, the present invention has developed a preservation method for an experimental population of Psylla pyrisuga. During the peak period of Psylla pyrisuga occurrence, a large number of Psylla pyrisuga eggs and 1st instar nymphs collected in the orchard are cultured into winter-type adults, which can achieve long-term preservation. Summary of the Invention

[0003] To solve the technical problems existing in the prior art, the present invention provides a preservation method for an experimental population of Psylla pyrisuga suitable for long-term preservation of Psylla pyrisuga in the laboratory.

[0004] To achieve the above object, the technical solution adopted by the present invention is a preservation method for an experimental population of Psylla pyrisuga, which is operated according to the following steps.

[0005] s1. Collect Psylla pyrisuga eggs and 1st instar nymphs in the orchard;

[0006] s2. Cut a hole with a diameter of 0.2 - 0.5 cm at the bottom of a centrifuge tube, insert the petiole of a fresh pear leaf into the hole, and make the petiole extend to two-thirds of the distance from the bottom of the centrifuge tube to the tube mouth. Leave 0.5 cm of the petiole outside the centrifuge tube, then fix the petiole outside and the centrifuge tube with a sealing film, and seal the gap between the petiole and the small hole. Finally, inject Hoagland solution into the centrifuge tube and put on the lid;

[0007] s3. Gently transfer the collected Psylla pyrisuga eggs and 1st instar nymphs to the pear leaf in step s2 with a fine brush. There are about 20 Psylla pyrisuga on each pear leaf, and then place the centrifuge tube in an enamel square plate and seal the enamel square plate with a plastic wrap;

[0008] s4. Place the enamel square plate in a climate chamber. When the Hoagland solution is consumed by more than half, refill it in time. When the pear leaf turns yellow and wilts, replace the leaf in time and transfer the Psylla pyrisuga until the Psylla pyrisuga develops into winter-type adults;

[0009] S5. Take several pear tree branches about 10 cm long after removing branches and leaves. The diameter of the branches is less than 1.5 cm, and dry the branches under natural light conditions for 24 h;

[0010] S6. Prepare the nutrient solution for Psylla pyrisuga;

[0011] S7. Immerse the nano-cotton square in the nutrient solution for Psylla pyrisuga. After sufficient soaking, take it out and squeeze out the water until the two planes are longitudinally squeezed to a gap of 0.5 cm without water leakage;

[0012] S8. Put 5-7 pear tree branches in step S5, 3-5 nano-cotton squares in step S7 and about 200 winter-type adults in step S4 into a 500 ml preservation bottle, and preserve them under the condition of 2℃ without light;

[0013] S9. During the preservation period, take it out every 15 days, replace the nano-cotton square in step S7, cover the bottle mouth with a 1000-mesh gauze, expose it to ultraviolet light at 20℃ for 10 minutes and place it under natural light at 20℃ for 4 hours, and then re-preserve it.

[0014] Preferably, in step S2, the Hoagland solution is made into a 2000-fold solution and then injected into a centrifuge tube.

[0015] Preferably, in step S4, the enamel square plate is placed in a climate chamber at a temperature of 20±1℃ and a photoperiod of L:D = 8h:16h.

[0016] Preferably, the preparation method of the nutrient solution for Psylla pyrisuga in step S6 is as follows: Add 20 ml of fresh crisp pear juice, 15 g of leucine, 9.5 g of alanine, 9 g of glycine, 9 g of arginine, 8 g of lysine, and 1.5 g of potassium sorbate to 1200 ml of pure water, stir evenly, and then sterilize it at high temperature and high pressure to make the nutrient solution for Psylla pyrisuga.

[0017] Preferably, in step S8, the density of preserved Psylla pyrisuga is at least 1.8 cm3 / head.

[0018] Compared with the prior art, the present invention has the following technical effects.

[0019] 1. By controlling the temperature and photoperiod during the development of Psylla pyrisuga eggs and 1st instar nymphs, the present invention enables them to develop into winter-type Psylla pyrisuga with stronger tolerance. Then, these winter-type Psylla pyrisuga are put into a bottle together with the pear tree branches after removing branches and leaves and the nano-cotton soaked with nutrient solution to supplement energy for Psylla pyrisuga, and are preserved and inspected and updated regularly, realizing the long-term preservation of Psylla pyrisuga in the laboratory.

[0020] 2. During the outbreak period of Psylla pyrisuga, eggs and 1st instar nymphs of Psylla pyrisuga were collected, solving the problem of difficult acquisition of Psylla pyrisuga and reducing the waste caused by the inability to test or process the test insects in time after outdoor collection.

[0021] 3. Utilizing the characteristics that the winter-type Psylla pyrisuga has weak activity and can survive for a long time under low temperature and no light, and can mate and reproduce after being transferred to normal temperature and photoperiod for a period of time, it is possible to obtain winter-type Psylla pyrisuga at any time and expand the population by controlling conditions to breed and obtain a sufficient amount of summer-type Psylla pyrisuga.

[0022] 4. The method of the present invention does not require the laboratory to always maintain a population of Psylla pyrisuga, avoiding long-term large-scale cultivation of pear tree seedlings and boring mechanical feeding processes. Detailed implementation manners

[0023] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clear and understandable, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0024] A preservation method for an experimental population of Psylla pyrisuga is operated according to the following steps.

[0025] s1. In the pear orchard during the outbreak period of Psylla pyrisuga, pick and preserve the leaves with a large number of eggs and 1st instar nymphs of Psylla pyrisuga and bring them back to the laboratory.

[0026] s2. Take a 5 ml centrifuge tube, cut a hole with a diameter of 0.2 - 0.5 cm at the bottom, insert the petiole of a fresh pear tree leaf into the hole, and make the petiole extend to two-thirds of the distance from the bottom of the centrifuge tube to the tube mouth. Leave 0.5 cm of the petiole outside the centrifuge tube, then fix the petiole outside and the centrifuge tube with a sealing film, and seal the gap between the petiole and the small hole. Finally, inject the 2000-fold solution prepared with Hoagland into the centrifuge tube and cover the lid.

[0027] s3. Gently transfer the collected eggs and 1st instar nymphs of Psylla pyrisuga to the pear tree leaf in step s2 with a fine brush. There are about 20 Psylla pyrisuga on each pear tree leaf, and then place the centrifuge tube in an enamel square tray and seal the enamel square tray with a plastic wrap.

[0028] s4. Place the enamel square tray in a climate chamber at a temperature of 20 ± 1°C and a photoperiod of L:D = 8h:16h. When the Hoagland solution is consumed by more than half, refill it in time. When the pear tree leaf turns yellow and wilts, replace the leaf in time and transfer the Psylla pyrisuga until the Psylla pyrisuga develops into winter-type adults.

[0029] s5. Take several pear tree branches about 10 cm long without branches and leaves, with a branch diameter less than 1.5 cm, and dry the branches under natural light conditions for 24 h.

[0030] S6. Prepare the nutrient solution for Psylla pyrisuga. The preparation method is as follows: Add 20 ml of freshly squeezed crisp pears, 15 g of leucine, 9.5 g of alanine, 9 g of glycine, 9 g of arginine, 8 g of lysine, and 1.5 g of potassium sorbate into 1200 ml of pure water. After stirring evenly, sterilize it by high temperature and high pressure to make the nutrient solution for Psylla pyrisuga;

[0031] S7. Immerse the nano-cotton square in the nutrient solution for Psylla pyrisuga. After sufficient immersion, take it out and squeeze out the water until the two planes are longitudinally squeezed to a gap of 0.5 cm without water leakage;

[0032] S8. Put 5 - 7 branches of pear tree branches in step S5, 3 - 5 nano-cotton squares in step S7, and about 200 winter-type adults in step S4 into a 500 ml preservation bottle, and preserve them under the condition of 2°C without light. The preservation density of Psylla pyrisuga is at least 1.8 cm3 / head;

[0033] S9. During the preservation period, take it out every 15 days, replace the nano-cotton square in step S7, cover the bottle mouth with a 1000-mesh gauze, expose it to ultraviolet light at 20°C for 10 minutes, place it at 20°C under natural light for 4 hours, and then preserve it again.

[0034] The above are only the preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present invention shall be included within the scope of the present invention.

Claims

1. A preservation method for an experimental population of Psylla pyrisuga, characterized in that: Operate according to the following steps: S1. Collect the eggs and 1st instar nymphs of Psylla pyrisuga in the pear orchard; S2. Cut a hole with a diameter of 0.2 - 0.5 cm at the bottom of a centrifuge tube, insert the petiole of a fresh pear tree leaf into the hole, and extend the petiole into the centrifuge tube to two-thirds of the distance from the tube mouth. Leave 0.5 cm of the petiole outside the centrifuge tube. Then fix the petiole outside and the centrifuge tube with sealing film, and seal the gap between the petiole and the small hole. Finally, inject Hoagland solution into the centrifuge tube and cover it with a lid; S3. Transfer the collected eggs and 1st instar nymphs of Psylla pyrisuga to the pear tree leaves in step S2. There are about 20 Psylla pyrisuga on each pear tree leaf. Then place the centrifuge tube in an enamel square tray and seal the enamel square tray with plastic wrap; S4. Place the enamel square tray in a climate chamber. When the Hoagland solution is consumed by more than half, refill it in time. When the pear tree leaves turn yellow and wither, replace the leaves in time and transfer the Psylla pyrisuga until the Psylla pyrisuga develop into winter-type adults; S5. Take several pear tree branches about 10 cm long without branches and leaves. The diameter of the branches is less than 1.5 cm, and dry the branches under natural light for 24 h; S6. Prepare the nutrient solution for Psylla pyrisuga; S7. Immerse the nano-cotton square in the nutrient solution for Psylla pyrisuga. After full immersion, take it out and squeeze out the water. Squeeze the two planes longitudinally until the gap is 0.5 cm and no water comes out; S8. Put 5 - 7 pear tree branches in step S5, 3 - 5 nano-cotton squares in step S7, and about 200 winter-type adults in step S4 into a 500 ml preservation bottle and preserve them at 2℃ in the dark; S9. During the preservation period, take it out every 15 days, replace the nano-cotton square in step S7, cover the bottle mouth with a 1000-mesh gauze, expose it to ultraviolet light at 20℃ for 10 minutes with the mouth open, and place it under natural light at 20℃ for 4 hours before re-preserving; 2. The preservation method of an experimental population of Psylla pyrisuga according to claim 1, characterized in that: In step S2, the Hoagland solution is made into a 2000-fold solution and then injected into the centrifuge tube.

3. A preservation method for an experimental population of Psylla pyrisuga according to claim 1, characterized in that: In step S4, place the enamel square tray in a climate chamber at a temperature of 20 ± 1℃ and a photoperiod of L:D = 8h:16h.

4. A preservation method for an experimental population of Psylla pyrisuga according to claim 1, characterized in that: The preparation method of the nutrient solution for Psylla pyrisuga in step S6 is as follows: Add 20 ml of freshly squeezed crisp pear juice, 15 g of leucine, 9.5 g of alanine, 9 g of glycine, 9 g of arginine, 8 g of lysine, and 1.5 g of potassium sorbate to 1200 ml of pure water, stir evenly, and then sterilize it by high temperature and high pressure to make the nutrient solution for Psylla pyrisuga.

5. The preservation method of an experimental population of Psylla pyrisuga according to claim 1, characterized in that: In the step s8, the density of preserved Psylla pyrisuga is at least 1.8 cm 3 / head.