CN111264320A - Method for accurately identifying resistance of brown planthopper in adult stage of rice - Google Patents
Method for accurately identifying resistance of brown planthopper in adult stage of rice Download PDFInfo
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- 241001556089 Nilaparvata lugens Species 0.000 title claims abstract description 72
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 43
- 235000009566 rice Nutrition 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 33
- 240000007594 Oryza sativa Species 0.000 title abstract 2
- 241000196324 Embryophyta Species 0.000 claims abstract description 47
- 241000238631 Hexapoda Species 0.000 claims abstract description 17
- 238000011156 evaluation Methods 0.000 claims abstract description 16
- 230000008654 plant damage Effects 0.000 claims abstract description 6
- 241000209094 Oryza Species 0.000 claims description 45
- 230000006378 damage Effects 0.000 claims description 25
- 239000000463 material Substances 0.000 claims description 17
- 241000607479 Yersinia pestis Species 0.000 claims description 9
- 239000004033 plastic Substances 0.000 claims description 5
- 238000009331 sowing Methods 0.000 claims description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 abstract description 8
- 238000011081 inoculation Methods 0.000 abstract description 6
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- 235000015001 Cucumis melo var inodorus Nutrition 0.000 description 2
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- 238000012360 testing method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000239290 Araneae Species 0.000 description 1
- 206010063659 Aversion Diseases 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
- A01G22/22—Rice
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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Abstract
The invention discloses a method for accurately identifying the resistance of brown planthoppers in the adult stage of rice. And (3) after 50 days of transplanting, assembling an iron bracket and a nylon net cover to identify plant cages, inoculating the number of the primary emergence female imagoes of the robust brown planthoppers according to 1 tillering of each plant, and taking the plant damage degree as a resistance evaluation index and the number of insect mouths as a reference index. The method is based on artificial inoculation identification, can be carried out in places such as greenhouses, artificial climates and the like, can effectively control environmental factors such as temperature, humidity, illumination and the like, provides stable and uniform identification conditions, and ensures the accuracy of identification results.
Description
Technical Field
The invention belongs to the technical field of agriculture, and particularly relates to a method for accurately identifying resistance of brown planthopper in a rice adult stage.
Background
Brown planthopper (Brown planthopper, Nilaparvata lugens)
Identification of resistance in screening Rice resistanceThe brown planthopper resource, the research and the utilization of the brown planthopper resistance gene and the like are indispensable key links. The identification method of brown planthopper resistance of rice mainly comprises field natural identification and artificial inoculation identification. At present, the brown planthopper resistance identification of rice is carried out by artificial inoculation in most of researches. The artificial inoculation identification is mainly divided into seedling stage identification and adult stage identification according to the identification period, and can be divided into single plant resistance identification and colony resistance identification according to the experimental purpose and the identification scale, wherein the insect resistance indexes comprise brown planthopper honeydew excretion amount, brown planthopper egg laying amount, insect population density, identification plant damage degree and the like.
However, existing identification methods often have a number of drawbacks:
the existing field natural identification scheme can not effectively control the period and scale of insect pests in the natural environment, and often causes the failure of experiments due to inaccurate identification results because of the problem of the insect source of the brown planthopper or the occurrence of local insect pests. The artificial inoculation identification is beneficial to manufacturing a stable experimental environment and obtaining a better identification effect.
At present, no research result clearly shows that the rice has consistent resistance to the brown planthopper at the seedling stage and the adult stage, most of the current researches carry out the identification of the resistance to the brown planthopper at the seedling stage of the rice, and the brown planthopper is mainly generated at the adult stage of the rice in the actual production, so the identification result of the resistance to the brown planthopper at the adult stage of the rice is more favorable for guiding the field production.
In the excavation process of the brown planthopper resistant gene of the rice, a segregation population is usually required to be constructed for gene localization. In most studies, when evaluating the resistance level of an isolated individual plant in a positioned group, the resistance level of the isolated individual plant is usually reversely deduced by performing group resistance identification on a next generation family of the isolated individual plant, so that the operation is complicated, the workload is large, and the time is long.
The resistance level of a rice plant is evaluated by identifying a single tillering in a partial identification method (such as a rice stem and stump identification method), but the tillering of a single rice plant is different from that of the whole plant in the aspects of growth vigor and the like, and the tillering is inevitably damaged in the peeling process, so that the physiological function of the tillering is damaged, and the resistance level of the whole plant is hardly reflected by the identification result of the single tillering.
In part of methods, identification materials are collected together for identification, unified inoculation is carried out according to the total quantity of the identification materials, and it is difficult to ensure that each plant of identification materials has a consistent quantity of brown planthoppers for feeding. The feeding tendency of the brown planthopper on the pest-susceptible plants is certainly stronger than that of the resistant plants, so that the brown planthopper after being inoculated with the pests can be greatly concentrated on the pest-susceptible plants to feed, the feeding frequency on the resistant plants can be greatly reduced, and the situation can hardly accurately distinguish the medium-resistant, resistant and high-resistant plants, thereby influencing the accuracy of resistance evaluation results.
The resistance level of rice to brown planthopper should be evaluated by combining the damage performance of rice under the harm of brown planthopper and the feeding behavior of brown planthopper group on rice, and most of the existing identification methods adopt a single identification index. The resistance mechanism of rice to brown planthopper is mainly divided into evasion, resistance and tolerance, and the resistance mechanism is difficult to be accurately distinguished by a single identification index. Taking the damage degree of the plant and the population density of the brown planthopper as examples, the plant with the resistance mechanism as the aversion shows that the damage degree is extremely light or not damaged, and the population density is extremely small or no insects; the damage degree is light on the plants with resistance mechanism as the antibiotic, and the population density is small; on the other hand, the plants with the resistance mechanism of tolerance may show a low degree of damage and a high population density. In this case, if the population density, honeydew excretion amount, egg excretion amount, and the like are used as identification indexes, a plant whose resistance mechanism is tolerance may be evaluated as an insect-susceptible plant.
Disclosure of Invention
The invention aims to provide a method for accurately identifying the resistance of brown planthoppers in the adult stage of rice, which takes the number of insect population as a reference index, considers the conditions of two aspects of plant expression and behavior of the brown planthoppers, judges the accuracy of an identification result through the correlation between the plant expression and the behavior of the brown planthoppers, is favorable for eliminating the interference of adverse factors and obtains an accurate identification result.
The invention is realized by the following technical scheme:
a method for accurately identifying the resistance of brown planthopper in the adult stage of rice comprises the following steps:
1) respectively selecting pest-susceptible control varieties and pest-resistant control varieties;
2) synchronously sowing the pest-sensing control, the pest-resistant control and the identification material in a field;
3) transplanting the three leaves into plastic pots in the one-heart stage, transplanting 2-3 plants in each pot, thinning seedlings after the normal growth of the striking root is recovered, and reserving a robust plant in each pot for identification;
4) after 50 days of transplanting, assembling a bracket and a net cover to cover the identified plants;
5) inoculating the number of the primary emergence female imagoes of the robust brown planthoppers according to 1 tillering of each branch;
6) and (3) judging the accuracy of the resistance evaluation result according to the correlation between the damage degree and the population number by taking the plant damage degree as a resistance evaluation index and taking the population number as a reference index.
Further, the pest-susceptible control is rice variety TN 1; the insect-resistant control is rice variety RathuHeenati (RHT).
Further, the resistance evaluation indexes are as follows:
further, when the pest-susceptible control reaches 7 grades, the number of the mouths of the brown planthopper adults and nymphs on the plants is investigated and identified, and when the pest-susceptible control reaches 9 grades, resistance evaluation is carried out according to the damage degree of the identified plants.
The invention discloses a special identification cage which comprises a plastic bowl, a cylindrical iron bracket and a cylindrical 40-mesh nylon mesh enclosure.
The brown planthopper insect source is collected in natural field population, and is bred and propagated on an insect-susceptible rice variety TN1 in an artificial climate chamber (the temperature is 30 ℃ and the humidity is about 90 percent).
The pest-sensing contrast, the pest-resistant contrast and the identification material of the invention are subjected to conventional water and fertilizer management in the whole growth period, and no pesticide is applied.
The invention has the beneficial effects that:
1) the invention establishes the method for identifying the resistance of the brown planthopper in the adult stage of the rice, can accurately control the number of the inoculated insects and control the migratory flight of the brown planthopper by identifying the single plant and the single cage, so that the brown planthopper can be fed and propagated on the identified plant, the condition that the brown planthopper is harmful to the rice in the natural environment is simulated, and the identification result is real and reliable. The method provides a high-efficiency and accurate resistance identification technology for screening of brown planthopper resistant resources of rice and mining of brown planthopper resistant genes, and can effectively promote the screening of the brown planthopper resistant resources of rice and the mining of the resistant genes.
2) The method takes the whole rice plant as a unit to identify the resistance of the brown planthopper in the adult plant period, and the resistance coincides with the main period of the brown planthopper harmful rice in the actual production in the growth period, so that the method is favorable for investigating the actual performance of the rice on the resistance of the brown planthopper in the field production and is favorable for guiding the field production.
3) The method can identify stable strain materials and also can identify separated individual strain materials. In the process of carrying out brown planthopper resistance gene excavation by using a rice low-generation segregating population, an evaluation result obtained by carrying out resistance identification on the segregating single plant through the method can be directly used for gene localization research, population identification on a next generation family of the segregating single plant is not needed any more to reversely deduce the resistance of the segregating single plant, time and labor are saved, and the gene excavation efficiency is improved.
4) The identification method takes the damage degree of the plant as an identification index, takes the population number as a reference index, considers the damage performance of the identified plant and the feeding and breeding behaviors of the brown planthopper, judges the accuracy of the identification result through the correlation between the damage performance and the feeding and breeding behaviors, is favorable for eliminating the adverse interference, obtains the accurate identification result and improves the gene positioning efficiency.
5) When the method is used for screening brown planthopper-resistant resources of rice, the resistance mechanism of the resistant resources can be preliminarily explored and judged according to the relationship between the plant damage degree and the population number, and the obtained result is helpful for developing further research and application work.
Drawings
FIG. 1 is a customized, dedicated authentication cage of the present invention, and a large-scale authentication;
FIG. 2 shows the identification of rice F according to the invention2Isolating brown planthopper resistance of the individual plants;
FIG. 3 shows the rice brown planthopper resistance QTL location of the present invention;
FIG. 4 is a comparison of the resistance to Nilaparvata lugens for R900-1 and R900 of the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to specific embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment provides a method for accurately identifying the resistance of brown planthopper in the adult stage of rice, which comprises the following steps:
an identification device: as shown in fig. 1, a dedicated identification cage is custom-made, which comprises a plastic bowl (diameter 25cm, height 30cm), a cylindrical iron bracket (diameter 23cm, height 140cm) and a cylindrical 40-mesh nylon mesh enclosure (diameter 24cm, height 140cm, lower portion with 40cm opening for magic tape).
The insect breeding scheme comprises the following steps: the brown planthopper insect source is collected in natural field population, and is bred and propagated on insect-susceptible rice variety TN1 in an artificial climate chamber (the temperature is 30 ℃ and the humidity is about 90%).
Identification scheme: the rice variety TN1 was used as a pest-susceptible control, and Rathu Heenati (RHT) was used as a pest-resistant control. The contrast and identification materials are sown in the field at the same time, the three leaves are moved into plastic pots in one heart period, 2-3 plants are transplanted in each pot, after the green turning is recovered to normal growth, the seedlings are thinned, and a robust plant is left in each pot for identification. And after 50 days of transplanting, assembling an iron bracket and a nylon net cover to cover the identified plant, and removing other organisms such as spider and the like in the identification device. And then inoculating the number of the primary emergence female imagoes of the robust brown planthoppers according to 1 tillering of each branch, checking the survival condition of the brown planthoppers after inoculating the insects for 2-3 days, and supplementing the insect quantity according to the actual condition. All materials are subjected to conventional water and fertilizer management in the whole growth period, and no pesticide is applied.
And (3) judging the accuracy of the resistance evaluation result according to the correlation between the damage degree and the population number by taking the plant damage degree as a resistance evaluation index and taking the population number as a reference index. And when the control TN1 reaches the grade 7, investigating and identifying the population number of the brown planthopper adults and nymphs on the plant, and when the control TN1 reaches the grade 9, performing resistance evaluation according to the damage degree of the identified plant. The correlation between the population number and the damage degree can be used for judging the accuracy of the evaluation result.
The resistance evaluation criteria using the degree of damage as an identification index were as follows:
example 2 identification of Rice F2Isolation of individual plant Nilaparvata lugens resistance and Gene mapping
Experimental materials: obtaining a high-brown planthopper-resistant material R2426 through resistance identification and screening, and constructing F by using the R2426 and an insect-susceptible contrast TN12Segregating the population for resistance identification and gene mapping.
The experimental scheme is as follows: the brown planthopper used for the test is taken from a natural mixed population in the field and is propagated by using an insect-susceptible control TN 1. 545 parts of F were treated by the technical method of the present invention with TN1 as an insect-susceptible control2And (5) separating the individual plant for identification. Wherein the identification material and the control TN1 are sown in 2016 at 6-month and 16-day, transplanted at 7-month and 10-day, and all identification individuals and the control TN1 are inoculated at 9-month and 1-day.
And (3) identification result: the insect population of the identified individual was investigated when the damage degree of the control TN1 reached grade 7 at day 28 of 9 months, and the damage degree of the control TN1 reached grade 9 at day 25 of 10 months, and the damage degree of the identified individual was investigated. The results showed that significant differences in resistance were exhibited between the identified individuals (FIG. 2), and the correlation coefficient between the extent of damage and the number of worms in 546 of the identified lines was 0.799. In order to provide more reliable phenotype data for gene mapping work, screening is carried out in 546 identification strains, and 335 identification strains with higher correlation (the correlation coefficient is 0.918, and P is less than 0.001) are screened for gene mapping according to the insect quantity and the damage degree of the identification strains. 3 rice brown planthopper resistant QTLs were mapped on chromosome 1, 6 and 10, respectively (FIG. 3). The result comparison confirms that the gene located on the 6 th staining is the cloned brown planthopper resistance gene BPH32, so that the identification result obtained by the technical method is accurate and reliable.
Example 3 application in Rice parent Brown planthopper resistance improvement Process
Experimental materials: taking the brown planthopper resistant material R2426 as a donor parent and the restorer R900 as a receptor parent, carrying out auxiliary selection on BPH32 by combining a molecular marker in a backcross mode, and obtaining a BC3F4 generation homozygous stable strain R900-1 carrying BPH32 after multiple generations.
The experimental scheme is as follows: the brown planthopper used for the test is taken from a natural mixed population in the field and is propagated by using an insect-susceptible control TN 1. The resistance of the brown planthopper of R900-1 is identified by the technical method of the invention by taking TN1 as an insect-susceptible control. Wherein the identification material and control TN1 are sown in 2019 at 5-20 days, transplanted at 6-10 days, and inoculated at 7-31 days.
And (3) identification result: the insect population of the identified material was investigated when the damage degree of the 9-month and 11-day control TN1 reached the 7-grade, and the damage degree of the 10-month and 4-day control TN1 reached the 9-grade, and the damage degree of the identified individual was investigated. The results showed that R900-1 carrying BPH32 showed significant resistance to brown planthopper, enabling normal heading and fructification, whereas the recipient parent R900 died under the harmfulness of brown planthopper (fig. 4).
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (4)
1. A method for accurately identifying the resistance of brown planthopper in the adult stage of rice is characterized by comprising the following steps:
1) respectively selecting pest-susceptible control varieties and pest-resistant control varieties;
2) synchronously sowing the pest-sensing control, the pest-resistant control and the identification material in a field;
3) transplanting the three leaves into plastic pots in the one-heart stage, transplanting 2-3 plants in each pot, thinning seedlings after the normal growth of the striking root is recovered, and reserving a robust plant in each pot for identification;
4) after 50 days of transplanting, assembling a bracket and a net cover to cover the identified plants;
5) inoculating the number of the primary emergence female imagoes of the robust brown planthoppers according to 1 tillering of each branch;
6) and (3) judging the accuracy of the resistance evaluation result according to the correlation between the damage degree and the population number by taking the plant damage degree as a resistance evaluation index and taking the population number as a reference index.
2. The method for accurately identifying the resistance of the brown planthopper at the adult stage of the rice as claimed in claim 1, wherein the pest-susceptible control is rice variety TN 1; the insect-resistant control is rice variety Rathu Heenati.
3. The method for accurately identifying the resistance of the brown planthopper at the adult stage of the rice as claimed in claim 1, wherein the resistance evaluation indexes are as follows:
4. the method for accurately identifying the resistance of the brown planthoppers in the adult stage of the rice as claimed in claim 1, wherein the population number of the brown planthopper adults and nymphs on the rice plant is investigated and identified when the pest-susceptible control reaches 7 grades, and the resistance evaluation is carried out according to the damage degree of the identified plant when the pest-susceptible control reaches 9 grades.
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| CN112243812A (en) * | 2020-08-27 | 2021-01-22 | 河南科技大学 | Wheat growth period field aphid-resistant screening and identifying device |
| CN114271158A (en) * | 2021-12-30 | 2022-04-05 | 华智生物技术有限公司 | Method for identifying resistance of brown planthopper of rice in field adult stage |
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Application publication date: 20200612 |