Evaluation bioassay method for resistance of poplar to anoplophora glabripennis

Abstract

The invention provides a bioassay method for evaluating resistance of poplar to anoplophora glabripennis, which comprises the following steps of: intercepting a branch of a first poplar variety through a non-selective method and a selective method; putting the cut branches into an experimental mesh cage, and inoculating first anoplophora glabripennis into the experimental mesh cage for feeding measurement; cutting wood segments of the first poplar variety through a non-selective method and a selective method; putting the cut wood sections into an experimental mesh cage, and connecting second anoplophora glabripennis into the experimental mesh cage for spawning measurement; third anoplophora glabripennis is inoculated into the forest greenhouse and the forest cage through the forest greenhouse method and the forest cage sleeving method in the emerging period of the anoplophora glabripennis; performing biological assay on the field anti-anoplophora glabripennis of the second poplar variety; the insect resistance of the poplar variety is researched by adopting different methods indoors and in the field, and then the insect resistance of the poplar variety is obtained by comprehensively evaluating the two methods. The determination method is simple to operate, good in repeatability and suitable for indoor and field trials.

Description

Technical Field

[0001] This invention relates to the field of plant insect resistance identification technology, and in particular to a bioassay method for evaluating the resistance of poplar to the longhorn beetle. Background Technology

[0002] The bare-shouldered star beetle Anoplophora glabripennis It is the most serious borer pest in my country's "Three-North" shelterbelt system. In the mid-1980s, the poplar longhorn beetle, mainly the Asian longhorn beetle (Aegilops glabripennis), caused a devastating infestation in Ningxia, resulting in the near-total loss of poplar trees, the main species in the shelterbelts, with over 80 million trees dying. In the early 1990s, it became rampant in Inner Mongolia. At the beginning of this century, it began to cause damage in Xinjiang, affecting more than 10 species of broad-leaved trees to varying degrees, with over 370,000 trees damaged. In recent years, the Asian longhorn beetle has wreaked havoc in the Hexi Corridor region, causing catastrophic damage to shelterbelt networks in many areas.

[0003] Poplar is a major afforestation tree species in the "Three-North" region of my country, possessing significant ecological and economic value, and is an irreplaceable protective forest species in Northwest China. Due to the damage caused by the Asian longhorn beetle, there is currently a severe shortage of available poplar species or varieties. Cultivating and promoting insect-resistant varieties is the most fundamental, economical, and effective means of controlling pests. Summary of the Invention

[0004] This invention provides a bioassay method for evaluating the resistance of poplar trees to the Asian longhorn beetle (A. glabra). The method involves studying the insect resistance of poplar varieties using different methods in both indoor and field settings, and then comprehensively evaluating the resistance of the poplar varieties using both methods. The feeding area and number of oviposition grooves of the Asian longhorn beetle on poplar varieties are then statistically analyzed to finally screen out the most resistant poplar varieties. The assay method of this invention is simple to operate, has good repeatability, and is suitable for both indoor and field trials.

[0005] This invention provides a bioassay method for evaluating the resistance of poplar to the longhorn beetle, comprising the following steps.

[0006] Branches of the first poplar variety were cut using both non-selective and selective methods. The cut branches were placed in an experimental net cage, and the feeding of the first Asian longhorn beetle was measured in the net cage. Tree segments of the first poplar variety were cut using both non-selective and selective methods. The cut tree segments were placed in an experimental net cage, and the second longhorn beetle was introduced into the experimental net cage to measure oviposition. During the peak emergence period of the *Anoplophora glabripennis*, a third *Anoplophora glabripennis* was introduced into the forest greenhouse and forest cage methods, respectively; bioassays were conducted on the field resistance of the second poplar variety to *Anoplophora glabripennis*.

[0007] According to the present invention, a bioassay method for evaluating the resistance of poplar to the longhorn beetle is provided. The first poplar varieties include Ta, Pa, Qinbaiyang No. 3, Northwest Poplar No. 2, and Xinjiang Poplar; the second poplar varieties include Pa, Qg, La, Ta, Ti, 107, I-101, Qinbaiyang No. 1, Qinbaiyang No. 2, Qinbaiyang No. 3, Qinbaiyang No. 4, Qinbaiyang No. 5, Qinbaiyang No. 7, Northwest Poplar No. 1, 2, 3, white poplar, and Xinjiang Poplar.

[0008] According to the present invention, a bioassay method for evaluating the resistance of poplar to the longhorn beetle is provided. In the non-selective method test, the experimental net cage is 1 m × 1 m × 1 m; in the selective test method test, the experimental net cage is 2 m × 2 m × 1.2 m.

[0009] According to the present invention, a bioassay method for evaluating the resistance of poplar to the longhorn beetle is provided, wherein the number of branches is three; the length of the branches is 40 cm and the diameter is 1 cm, and the branches are attached with leaves.

[0010] According to the present invention, a bioassay method for evaluating the resistance of poplar longhorn beetle *Aegilops glabripennis* is provided. The first *Aegilops glabripennis* includes female and male adults. In the non-selective method test, the number of the first *Aegilops glabripennis* is 2. In the selective method test, the number of the first *Aegilops glabripennis* is 10.

[0011] According to the present invention, a bioassay method for evaluating the resistance of poplar to the longhorn beetle is provided, wherein the height of the wood segment is 60 cm and the diameter is 10 cm.

[0012] According to the present invention, a bioassay method for evaluating the resistance of poplar longhorn beetle *Aegilops glabripennis* is provided. The second *Aegilops glabripennis* includes female and male adults. In the non-selective method test, the number of second *Aegilops glabripennis* is 1 male and 3 females. In the selective method test, the number of second *Aegilops glabripennis* is 2 males and 5 females.

[0013] The bioassay method for evaluating the resistance of poplar to the longhorn beetle provided by the present invention further includes: a second poplar variety being a cutting with two years of roots and one year of stem.

[0014] According to the present invention, a bioassay method for evaluating the resistance of poplar trees to the longhorn beetle *Pterocarya stenoptera* is provided. The forest greenhouse is a rectangular test area with a length of 13.7 m, a width of 15.2 m, and a height of 4.5 m, enclosed by wire mesh and iron frames. Poplar trees of a second variety are transplanted into the forest greenhouse in a randomized block design, with 5 trees of each variety. The spacing between two adjacent poplar trees is 50 cm. In the experiment of introducing the third-shouldered longhorn beetle into a forest greenhouse, the number of beetles was 100 males and 200 females.

[0015] According to the present invention, a bioassay method for evaluating the resistance of poplar to the longhorn beetle 'Spotted Longhorn Beetle' further includes: in the experiment of introducing the third longhorn beetle 'Spotted Longhorn Beetle' into the forest cage, the second poplar variety is transplanted in a continuous arrangement of two varieties per row, five trees of each variety, with a spacing of 50 cm between each tree. The cover of the forest cage is made of 100-mesh netting with a length of 70 cm and a width of 45 cm; the two ends of the forest cage are tied with wire, and the forest cage is placed on the trunk of each tree at a height of 1.5m above the ground. In the experiment of introducing the third-shouldered longhorn beetle into cages in the forest, the third-shouldered longhorn beetle was an adult, with 1 male and 2 females.

[0016] This invention provides a bioassay method for evaluating the resistance of poplar trees to the longhorn beetle (Anoplophora glabripennis). The method involves using both non-selective and selective methods to collect branches from a first poplar variety. These branches are placed in experimental net cages, and the first longhorn beetle is introduced into the cages for feeding measurements. Similarly, using non-selective and selective methods, logs of the first poplar variety are collected. These logs are placed in experimental net cages, and the second longhorn beetle is introduced into the cages for oviposition measurements. During the peak emergence period of the longhorn beetle, a third longhorn beetle is introduced into the forest greenhouse and forest cages, respectively, using both methods. Bioassays are then performed on the second poplar variety's resistance to the longhorn beetle in the field. By studying the insect resistance of poplar varieties using different methods in both indoor and field settings, the insect resistance of the poplar varieties is comprehensively evaluated using both methods. Finally, the feeding area and the number of oviposition grooves on the poplar varieties are statistically analyzed to select the most resistant poplar varieties. The measurement method of the present invention is simple to operate, has good repeatability, and is suitable for indoor and field trials. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0018] Figure 1 This is a schematic flowchart of the bioassay method for evaluating the resistance of poplar to the longhorn beetle provided by the present invention.

[0019] Figure 2 This is a test diagram of a poplar forest greenhouse resistant to the longhorn beetle, provided by the present invention.

[0020] Figure 3The results of field bioassays on the resistance of poplar varieties to the longhorn beetle provided by this invention are the number of oviposition grooves in the forest greenhouse.

[0021] Figure 4 The results of field bioassays on the poplar variety resistant to the longhorn beetle provided by this invention show the number of oviposition grooves in the forest cages. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0023] The following is combined with Figures 1-4 This invention is described.

[0024] Figure 1 This is a schematic flowchart of the bioassay method for evaluating the resistance of poplar to the longhorn beetle provided by the present invention, as shown in the figure. Figure 1 As shown, the method includes the following: Step 101: Using non-selective and selective methods, cut branches of the first poplar variety; place the cut branches into an experimental net cage, and introduce the first longhorn beetle (Aegilops glabripennis) into the experimental net cage for feeding measurement.

[0025] Specifically, the indoor determination of the resistance of poplar varieties to feeding behavior of the longhorn beetle was conducted using both non-selective and selective methods. Branches of poplar varieties were cut and placed in experimental net cages to be fed to the longhorn beetle for feeding determination.

[0026] Step 102: Using non-selective and selective methods, cut logs of the first poplar variety; place the cut logs into an experimental net cage, and introduce a second longhorn beetle (Aegilops glabripennis) into the experimental net cage to measure its egg-laying.

[0027] Specifically, the indoor determination of the resistance of poplar varieties to oviposition behavior of the longhorn beetle was conducted using non-selective and selective methods. Poplar tree segments were cut and placed in experimental net cages to be incubated with the longhorn beetle for oviposition determination.

[0028] Step 103: Using the forest greenhouse method and the forest cage method, during the peak emergence period of the *Anoplophora glabripennis*, the third *Anoplophora glabripennis* was introduced into the forest greenhouse and the forest cage respectively; bioassays were conducted on the field resistance of the second poplar variety to *Anoplophora glabripennis*.

[0029] Specifically, the field bioassay for the resistance of poplar varieties to the longhorn beetle *Anoplophora glabripennis* was conducted using greenhouses and cages within the forest. During the peak emergence period of *Anoplophora glabripennis*, the beetles were introduced into the greenhouses and cages to perform bioassays on the field resistance of poplar varieties to *Anoplophora glabripennis*. For example... Figure 2 As shown, Figure 2 Test diagram of a greenhouse for poplar trees resistant to the light-resistant longhorn beetle.

[0030] Optionally, the first poplar varieties include Ta, Pa, Qinbaiyang No. 3, Northwest Poplar No. 2, and Xinjiang Poplar; the second poplar varieties include Pa, Qg, La, Ta, Ti, 107, I-101, Qinbaiyang No. 1, Qinbaiyang No. 2, Qinbaiyang No. 3, Qinbaiyang No. 4, Qinbaiyang No. 5, Qinbaiyang No. 7, Northwest Poplar No. 1, 2, 3, White Poplar, and Xinjiang Poplar.

[0031] Optionally, in non-selective method tests, the experimental net cage is 1 m × 1 m × 1 m; in selective method tests, the experimental net cage is 2 m × 2 m × 1.2 m.

[0032] Specifically, in steps 101 and 102, the experimental net cage in the non-selective test is 1 m × 1 m × 1 m, and the experimental net cage in the selective test is 2 m × 2 m × 1.2 m.

[0033] Optionally, the number of branches is three; the branches are 40 cm long and 1 cm in diameter, and the branches are accompanied by leaves.

[0034] Specifically, the branches are about 40 cm long and about 1 cm in diameter, and each branch has leaves attached; there are three branches in total.

[0035] Optionally, the first-shouldered Asian longhorn beetle includes female and male adults; in the non-selective method test, the number of first-shouldered Asian longhorn beetles is 2; in the selective method test, the number of first-shouldered Asian longhorn beetles is 10.

[0036] Specifically, step 101 includes the feeding behavior of female adults and the feeding behavior of male adults. Two adults are introduced in the non-selective experiment and ten adults are introduced in the selective experiment.

[0037] Optionally, the log segment is 60 cm high and 10 cm in diameter.

[0038] Specifically, in step 102, the height of the wooden segment is approximately 60 cm and the diameter is approximately 10 cm.

[0039] According to the present invention, a bioassay method for evaluating the resistance of poplar longhorn beetle *Aegilops glabripennis* is provided. The second *Aegilops glabripennis* includes female and male adults. In the non-selective method test, the number of second *Aegilops glabripennis* is 1 male and 3 females. In the selective method test, the number of second *Aegilops glabripennis* is 2 males and 5 females.

[0040] Specifically, in step 102, one male and three female insects are introduced in the non-selective test, and two males and five female insects are introduced in the selective test.

[0041] Optionally, the bioassay method for evaluating the resistance of poplar to the longhorn beetle provided by the present invention further includes: a second poplar variety being a cutting with two years of roots and one year of stem.

[0042] Specifically, in step 103, all poplar varieties in the field were cuttings with two-year-old roots and one-year-old trunks.

[0043] Optionally, the forest greenhouse is a rectangular experimental area enclosed by wire mesh and iron frames, with a length of 13.7 m, a width of 15.2 m, and a height of 4.5 m; and each poplar of the second poplar variety is transplanted into the forest greenhouse in a randomized block arrangement of 5 trees of each variety; the spacing between two adjacent poplar trees is 50 cm; In the experiment of introducing the third-shouldered longhorn beetle into a forest greenhouse, the number of beetles was 100 males and 200 females.

[0044] Specifically, in step 103, the forest greenhouse is a rectangular experimental area measuring 13.7 m × 15.2 m × 4.5 m, enclosed by wire mesh and iron frames. Poplar trees of each variety are transplanted into the greenhouse in a randomized block design, with a spacing of 50 cm between adjacent trees. During the peak emergence period of the longhorn beetle, 100 males and 200 females are introduced into the greenhouse to measure oviposition.

[0045] Optionally, the present invention provides a bioassay method for evaluating the resistance of poplar to the longhorn beetle, which further includes: in the experiment of introducing the third longhorn beetle into the cage in the forest, the second poplar variety is transplanted in a continuous arrangement of two varieties per row, five trees of each variety, with a spacing of 50 cm between each tree.

[0046] Specifically, in step 103, the poplar varieties in the forest cages are transplanted in a continuous arrangement with 2 varieties per row, 5 trees of each variety, and 50cm between each tree.

[0047] Optionally, the cover of the forest cage is made of 100-mesh netting with a length of 70 cm and a width of 45 cm; the two ends of the forest cage are tied with wire, and the forest cage is placed on the trunk of each tree at a height of 1.5 m above the ground. In the experiment of introducing the third-shouldered longhorn beetle into cages in the forest, the third-shouldered longhorn beetle was an adult, with 1 male and 2 females.

[0048] Specifically, in step 103, the cover in the forest trap is made of 70 cm × 45 cm 100 mesh netting, with the ends tied with wire. It is placed on the trunk of each tree at a height of 1.5 m above the ground, and then one male and two female adult longhorn beetles are placed inside to conduct forced oviposition testing.

[0049] The feeding behavior of longhorn beetles directly affects tree health; their oviposition behavior influences the population density of the next generation and long-term damage. This invention, in indoor testing, not only measures the feeding behavior of longhorn beetles but also their oviposition behavior, comprehensively evaluating poplar resistance from the two most critical life stages of the pest. The results are more comprehensive and reliable. Compared to traditional indoor biological testing, this method better reflects the resistance traits of different varieties and treated plants to cutworms under natural conditions, and allows for on-site inoculation, observation of growth, and assessment of damage.

[0050] This invention combines indoor and field measurements. Indoor measurements, conducted under controlled conditions, effectively eliminate interference from complex variables in the field such as wind, rain, temperature, and predators. This allows for precise comparisons of the attraction or resistance of different tree varieties to longhorn beetles, resulting in scientifically rigorous and highly reproducible results. Field measurements, conducted under conditions closely resembling real forestry production, reflect the comprehensive resistance of trees in their actual growth environment, including the influence of tree vigor and microenvironment. This enables the research findings to be directly applied to field breeding and afforestation practices.

[0051] To further explain the present invention, the following Examples 1 to 3 are provided to illustrate the indoor determination of feeding behavior of poplar varieties resistant to the longhorn beetle, the indoor determination of oviposition behavior of poplar varieties resistant to the longhorn beetle, and the field bioassay of poplar varieties resistant to the longhorn beetle.

[0052] Example 1 Indoor determination of the feeding behavior of poplar varieties against the longhorn beetle: Using non-selective and selective methods, branches of poplar varieties were cut and placed in experimental net cages to be fed to the longhorn beetle for feeding determination.

[0053] Indoor testing of the resistance of poplar varieties to feeding behavior of the longhorn beetle was conducted using Ta, Pa, Qinbaiyang No. 3, Northwest Poplar No. 2, and Xinjiang Poplar.

[0054] The experimental net cage in the non-selective test was 1 m × 1 m × 1 m, and the experimental net cage in the selective test was 2 m × 2 m × 1.2 m.

[0055] Three poplar branches, each approximately 40cm long and 1cm in diameter, with leaves attached, were cut.

[0056] The indoor testing of feeding behavior included both female and male adults. In the non-selective experiment, branches of each poplar variety were inserted into glass bottles filled with water and then placed at the bottom center of experimental net cages for individual testing. Two adults were placed into each net cage and allowed to feed freely. New poplar branches and insects were replaced daily for five days, and the feeding area on the branches was measured at the end.

[0057] In the selective experiment, branches of each poplar variety were inserted into glass bottles filled with water and then arranged at equal intervals at the bottom of experimental net cages according to a completely randomized experimental design. Ten adult insects were then introduced into each net cage and allowed to feed freely. New poplar branches and insects were replaced daily for a total of 5 days. Finally, the feeding area on the branches was measured.

[0058] Analysis: One-way ANOVA was performed on the data using SPSS statistical software, and Tukey's test was used for multiple comparisons.

[0059] The results of indoor measurements of the feeding behavior of poplar varieties resistant to the longhorn beetle are shown in Table 1.

[0060] Table 1

[0061] The results showed that Qinbaiyang No. 3 and Northwest Poplar No. 2 had stronger resistance to the Asian longhorn beetle than other varieties.

[0062] Example 2 Indoor determination of the oviposition behavior of poplar varieties against the longhorn beetle: Using non-selective and selective methods, logs of poplar varieties were cut and placed in experimental net cages to be incubated with the longhorn beetle for oviposition determination.

[0063] Indoor testing of the oviposition behavior of poplar varieties against the longhorn beetle was conducted using Ta, Pa, Qinbaiyang No. 3, Northwest Poplar No. 2, and Xinjiang Poplar.

[0064] The experimental net cage in the non-selective test was 1 m × 1 m × 1 m, and the experimental net cage in the selective test was 2 m × 2 m × 1.2 m.

[0065] The section of poplar wood cut from the tree is approximately 60 cm high and 10 cm in diameter.

[0066] In the non-selective experiment, logs were placed at the bottom center of different experimental net cages according to species, and fresh branches were placed next to the logs to supplement the longhorn beetle's nutrition. Then, one male and three female beetles were introduced and allowed to lay eggs at will. After 10 days, the logs were removed, and the egg-laying grooves on the logs were recorded.

[0067] In the selective experiment, log segments were arranged at equal intervals at the bottom of the experimental net cage according to a completely randomized experimental design. Fresh branches were placed next to each log segment to supplement the longhorn beetle's nutrition. Then, two male beetles and five female beetles were introduced. They were allowed to lay eggs at will. After 10 days, the log segments were removed, and the egg-laying grooves on the log segments were recorded.

[0068] Analysis: One-way ANOVA was performed on the data using SPSS statistical software, and Tukey's test was used for multiple comparisons.

[0069] The results of indoor measurements of the oviposition behavior of poplar varieties against the longhorn beetle are shown in Table 2.

[0070] Table 2 species Non-selective test - oviposition groove Selective testing - egg-laying groove Ta 7.80 ± 1.07 a 12.20 ± 1.77 ab Pa 10.00 ± 2.17 a 20.00 ± 2.53 a Qin Baiyang No. 3 5.60 ± 1.21 ab 4.80 ± 0.93 b Northwest Poplar No. 2 2.00 ± 0.94 b 7.20 ± 2.87 b Xinjiang Yang 2.00 ± 1.10 b 8.40 ± 4.29 b The results showed that Populus chinensis 'Qinbaiyang No. 3', Populus 'Northwest' 'No. 2', and Populus 'Xinjiang' exhibited stronger resistance to the Asian longhorn beetle than other varieties.

[0071] Example 3 Bioassay of Poplar Varieties' Resistance to the Longhorn Beetle in the Field: Using the methods of forest greenhouses and forest cages, the longhorn beetles were introduced into the greenhouses and cages during the peak emergence period of the Longhorn Beetle in the forest, and bioassay of Poplar Varieties' Resistance to the Longhorn Beetle in the field was carried out.

[0072] Poplar varieties tested in the field included Pa, Qg, La, Ta, Ti, 107, I-101, Qinbaiyang No. 1, 2, 3, 4, 5, 7, Northwest Poplar No. 1, 2, 3, Maobaiyang, and Xinjiang Poplar. All poplar varieties tested in the field were cuttings with two-year-old roots and one-year-old trunks.

[0073] The experimental area, a rectangular enclosure measuring 13.7 m × 15.2 m × 4.5 m, was constructed using wire mesh and a metal frame. Five poplar trees of each variety were transplanted into the enclosure in a randomized block design, with a spacing of 50 cm between adjacent trees. During the peak emergence period of the longhorn beetle, 100 males and 200 females were introduced into the enclosure for a selective oviposition experiment. Ten days later, the oviposition grooves on the poplar trees were recorded.

[0074] Poplar varieties in the forest cages were transplanted in a continuous arrangement, with two varieties per row, five trees of each variety, and each tree spaced 50cm apart.

[0075] The traps used in the forest were made of 70 cm × 45 cm 100-mesh gauze netting, with the ends tied with wire. The traps were placed over the trunk of each tree at a height of 1.5 m above the ground. One male and two female adult longhorn beetles were then placed inside to conduct a forced oviposition experiment. After 10 days, the oviposition grooves on the poplar varieties were recorded.

[0076] Field bioassay results of resistance to the longhorn beetle in poplar varieties, such as... Figure 3 and Figure 4 As shown.

[0077] The results showed that in the forest greenhouse, the Qinbaiyang No. 2, Qinbaiyang No. 3, Qinbaiyang No. 7, Northwest Poplar No. 2, and 107 pairs of the Asian longhorn beetle exhibited stronger insect resistance than other varieties; in the forest cage trapping, the Qinbaiyang No. 1, Qinbaiyang No. 2, Qinbaiyang No. 3, Qinbaiyang No. 4, Qinbaiyang No. 5, Northwest Poplar No. 1, Northwest Poplar No. 3, White Poplar, and I-101 pairs of the Asian longhorn beetle exhibited stronger insect resistance than other varieties.

[0078] This invention provides a bioassay method for evaluating the resistance of poplar to the longhorn beetle *Aegilops glabripennis*. The method involves using both non-selective and selective methods to collect branches from a first poplar variety. These branches are placed in experimental net cages, and the first *Aegilops glabripennis* is introduced into the cages for feeding measurements. Similarly, using both non-selective and selective methods, logs of the first poplar variety are collected and placed in experimental net cages, where a second *Aegilops glabripennis* is introduced for oviposition measurements. During the peak emergence period of *Aegilops glabripennis*, a third *Aegilops glabripennis* is introduced into both the greenhouse and the cage-like structures in the forest. Bioassays are then performed on the second poplar variety to assess its resistance to *Aegilops glabripennis* in the field. By studying the insect resistance of poplar varieties using different methods in both indoor and field settings, the two methods are comprehensively evaluated to determine the overall insect resistance. Finally, the feeding area and number of oviposition grooves of *Aegilops glabripennis* on the poplar varieties are statistically analyzed to select the most resistant poplar varieties. The measurement method of the present invention is simple to operate, has good repeatability, and is suitable for indoor and field trials.

[0079] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A bioassay method for evaluating the resistance of poplar to the longhorn beetle, characterized in that, include: Branches of the first poplar variety were cut using both non-selective and selective methods. The cut branches were placed in an experimental net cage, and the feeding of the first Asian longhorn beetle was measured in the net cage. Tree segments of the first poplar variety were cut using both non-selective and selective methods. The cut tree segments were placed in an experimental net cage, and the second longhorn beetle was introduced into the experimental net cage to measure oviposition. During the peak emergence period of the *Anoplophora glabripennis*, a third *Anoplophora glabripennis* was introduced into the forest greenhouse and forest cage methods, respectively; bioassays were conducted on the field resistance of the second poplar variety to *Anoplophora glabripennis*.

2. The bioassay method for evaluating the resistance of poplar trees to the longhorn beetle according to claim 1, characterized in that, The first poplar varieties include Ta, Pa, Qinbaiyang No. 3, Northwest Poplar No. 2, and Xinjiang Poplar; the second poplar varieties include Pa, Qg, La, Ta, Ti, 107, I-101, Qinbaiyang No. 1, Qinbaiyang No. 2, Qinbaiyang No. 3, Qinbaiyang No. 4, Qinbaiyang No. 5, Qinbaiyang No. 7, Northwest Poplar No. 1, 2, 3, White Poplar, and Xinjiang Poplar.

3. The bioassay method for evaluating the resistance of poplar to the longhorn beetle according to claim 1, characterized in that, In the non-selective method test, the experimental net cage was 1 m × 1 m × 1 m; in the selective method test, the experimental net cage was 2 m × 2 m × 1.2 m.

4. The bioassay method for evaluating the resistance of poplar trees to the longhorn beetle according to claim 1, characterized in that, The number of branches is three; the branches are 40 cm long and 1 cm in diameter, and the branches are accompanied by leaves.

5. The bioassay method for evaluating the resistance of poplar to the longhorn beetle according to claim 1, characterized in that, The first *Aegilops glabripennis* includes female and male adults; in the non-selective method test, the number of the first *Aegilops glabripennis* is 2; in the selective method test, the number of the first *Aegilops glabripennis* is 10.

6. The bioassay method for evaluating the resistance of poplar to the longhorn beetle according to claim 1, characterized in that, The wooden segment is 60 cm high and 10 cm in diameter.

7. The bioassay method for evaluating the resistance of poplar trees to the longhorn beetle according to claim 1, characterized in that, The second *Aegilops glabripennis* includes female and male adults; in the non-selective method test, the number of the second *Aegilops glabripennis* is 1 male and 3 females; in the selective method test, the number of the second *Aegilops glabripennis* is 2 males and 5 females.

8. The bioassay method for evaluating the resistance of poplar to the longhorn beetle according to claim 2, characterized in that, Also includes: The second poplar variety is a cutting propagated from two-year-old roots and one-year-old trunk.

9. The bioassay method for evaluating the resistance of poplar to the longhorn beetle according to claim 2, characterized in that, The forest greenhouse is a rectangular experimental area enclosed by wire mesh and iron frames, with a length of 13.7 m, a width of 15.2 m, and a height of 4.5 m. Each poplar tree of the second poplar variety is transplanted into the forest greenhouse in a randomized block arrangement of 5 trees of each variety. The spacing between two adjacent poplar trees is 50 cm. In the experiment of introducing the third-shouldered longhorn beetle into a forest greenhouse, the number of the third-shouldered longhorn beetle was 100 males and 200 females.

10. The bioassay method for evaluating the resistance of poplar to the longhorn beetle according to claim 2, characterized in that, Also includes: In the experiment of introducing the third type of longhorn beetle into the forest cage, the second poplar variety was transplanted in a row of 2 varieties, with 5 trees of each variety, and each tree was spaced 50cm apart, in a continuous planting manner. The cover of the forest cage is made of 100-mesh netting with a length of 70 cm and a width of 45 cm; the two ends of the forest cage are tied with wire, and the forest cage is placed on the trunk of each tree at a height of 1.5m above the ground. In the experiment of introducing the third-shouldered longhorn beetle into cages in the forest, the third-shouldered longhorn beetle was an adult, and the number of the third-shouldered longhorn beetle was 1 male and 2 females.

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