Method for creating durable broad-spectrum brown planthopper-resistant rice material through polygene polymerization

Abstract

The invention discloses a method for creating a durable broad-spectrum brown planthopper-resistant rice material through polygene polymerization, and belongs to the technical field of rice molecular breeding. Indica type rice 9311 which is large in popularization area in production and excellent in comprehensive character is selected as a recurrent parent, a near-isogenic line material of seven resistance genes is constructed, then the near-isogenic lines of different genes are subjected to pairwise hybridization and complete diserial hybridization, molecular marker-assisted selection, a gene chip, a conventional breeding technology and brown planthopper resistance identification are combined, and the resistance of the brown planthopper is identified. According to the invention, four kinds of double-gene polymerized rice materials, namely BPH6 / BPH33, BPH14 / BPH33, BPH31 / BPH33 and BPH32 / BPH33, are totally obtained, and are novel rice materials with lasting broad-spectrum brown planthopper resistance. According to the strain bred by the method, the yield and the resistance are well coordinated, and the brown planthopper lasting broad-spectrum resistance is remarkably improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of crop molecular breeding, and relates to a method for creating a rice material with durable and broad-spectrum resistance to the brown planthopper by pyramiding multiple genes. Background Art

[0002] Rice (Oryza sativa L.) is a very important food crop, and the brown planthopper is one of the most destructive pests, causing huge losses to rice production. Traditional chemical control methods not only damage the environment but also enhance the drug resistance of the brown planthopper, which may lead to the resurgence of pests. Using endogenous rice genes resistant to the brown planthopper to breed insect-resistant varieties is the most economical and environmentally friendly control strategy. Due to the co-evolution of the brown planthopper and rice, varieties with resistance conferred by a single resistance gene are easily eliminated, while pyramiding multiple resistance genes can make rice varieties more broadly resistant to the brown planthopper and have a longer duration of resistance. However, when multiple genes are simultaneously pyramided into the same rice variety, the interaction effects between resistance genes are difficult to predict. Therefore, it is particularly important to accurately evaluate the interaction effects between resistance genes, and screening out resistance gene combinations with strong positive interaction effects is of great significance for the genetic improvement of rice resistance to the brown planthopper.

[0003] According to the different virulence capabilities of brown planthopper populations, their biotypes are generally divided into four types: biotype I, biotype II, biotype III, and biotype V. These biotypes differ in terms of virulence, adaptability, etc. For example, biotype I specifically feeds on and damages the rice variety Taichung No. 1 (TN1, which does not carry any insect-resistant genes); biotype II can feed on and threaten both TN1 and Mudgo (carrying the insect-resistant gene Bph1) rice varieties; biotype III also has the ability to feed on and damage both TN1 and ASD7 (carrying another insect-resistant gene Bph2) rice varieties; biotype V, as a specific physiological race in South Asia, has the most significant destructive power. The formation of biotypes is a very complex process, which is actually the result of the co-evolution of the brown planthopper and rice. The same brown planthopper population will have different virulence when living on different resistant rice varieties. Some scholars' research shows that the brown planthopper can quickly adapt to the resistance of rice, and generally can adapt to the resistance of insect-resistant rice after 10 generations.

[0004] Molecular marker-assisted selection technology (MAS) is efficient and precise in rice breeding and is widely used by researchers. By combining MAS with conventional breeding techniques, the efficient pyramiding of multiple genes in the same material can be achieved. The resistance spectra of individual resistance genes may vary. After gene pyramiding, the resistance spectra of the genes can be effectively broadened and the resistance level can be further enhanced, thus achieving the goal of durable broad-spectrum resistance. With the rapid development of rice functional genomics research, a large number of brown planthopper-resistant genes have been finely mapped and cloned, providing abundant molecular markers for MAS of genes, which can then be quickly used for the genetic improvement of insect-resistant varieties. Previously, we successfully constructed near-isogenic line (NILs) materials carrying multiple brown planthopper-resistant genes (BPH3, BPH6, BPH9, BPH14, BPH31, BPH32, and BPH33) based on the indica rice 9311 background. On this basis, by using MAS and conventional breeding methods, combined with insect resistance identification, four double-gene combination materials with durable broad-spectrum resistance to brown planthoppers and excellent agronomic traits were successfully obtained. Such improved lines can be used either as intermediate materials for insect resistance improvement or directly as new restorer line male parents to hybridize with excellent two-line sterile lines currently used in production to select new hybrid rice combinations with high resistance, high quality, and high yield, providing an effective solution to the problem of poor durable broad-spectrum insect resistance of current rice varieties in China. Summary of the Invention

[0005] The object of the present invention is to provide a method for creating a rice material with durable broad-spectrum resistance to brown planthoppers by pyramiding multiple genes.

[0006] To achieve the above object, the technical solution adopted by the present invention is as follows: The first aspect of the present invention provides a method for creating a rice material with durable broad-spectrum resistance to brown planthoppers by pyramiding multiple genes, comprising the following steps: (1) Using the homozygous insect-resistant near-isogenic lines BPH3-NIL, BPH6-NIL, BPH9-NIL, BPH14-NIL, BPH31-NIL, BPH32-NIL, and BPH33-NIL carrying a single insect-resistant gene under the rice 9311 background as the basic materials, performing pairwise hybridization and complete diallel crossing to obtain the F1 generation of 21 hybrid combinations; (2) After planting the F1 generation by plant line, using MAS molecular marker-assisted breeding technology to screen out F1 single plants with heterozygous resistance genotypes for the corresponding insect-resistant target genes, and bagging to harvest selfed seeds F2; (3) After planting the F2 generation by plant line, using MAS molecular marker-assisted breeding technology to screen out F2 single plants with homozygous resistance genotypes for the corresponding insect-resistant target genes, and bagging to harvest selfed seeds F3; (4) After planting the F3 generation of the ramet lines, based on the field agronomic traits and combined with the genetic background selection of the rice gene chip, F3 single plants with agronomic traits similar to those of indica rice 9311 and a genetic background similarity of more than 95% were screened out, and the selfed seeds F4 were harvested by bagging; (5) After planting the F4 generation of the ramet lines, selfing was carried out for three consecutive generations to obtain the F7 generation; The identification of the brown planthopper resistance characteristics showed that the F7 generations integrating BPH6 and BPH33, the F7 generations integrating BPH14 and BPH33, the F7 generations integrating BPH31 and BPH33, and the F7 generations integrating BPH32 and BPH33 were persistent broad-spectrum brown planthopper-resistant rice materials; Among them, BPH3 is a gene cluster composed of three genes with gene numbers LOC_Os04g12540, LOC_Os04g12560, and LOC_Os04g12580; The gene number of BPH6 is LOC_Os04g35210; BPH9 is a gene cluster composed of two genes with gene numbers LOC_Os12g37280 and LOC_Os12g37290; The gene number of BPH14 is LOC_Os03g63150; The gene number of BPH31 is LOC_Os11g29150; The gene number of BPH32 is LOC_Os06g03240; The gene number of BPH33 is LOC_Os04g02520; The molecular marker of BPH3 is H939, and the primers of molecular marker H939 are SEQ ID NO.1 - 2; The molecular marker of BPH6 is RM5757, and the primers of molecular marker RM5757 are SEQ ID NO.3 - 4; The molecular marker of BPH9 is H12, and the primers of molecular marker H12 are SEQ ID NO.5 - 6; The molecular marker of BPH14 is H14, and the primers of molecular marker H14 are SEQ ID NO.7 - 8; The molecular marker of BPH31 is 31 - 1, and the primers of molecular marker 31 - 1 are SEQ ID NO.9 - 10; The molecular marker of BPH32 is 32 - 5, and the primers of molecular marker 32 - 5 are SEQ ID NO.11 - 12; The molecular marker of BPH33 is H99, and the primers of molecular marker H99 are SEQ ID NO.13 - 14.

[0007] The second aspect of the present invention provides a molecular marker primer composition, which includes: primers for molecular marker H939, SEQ ID NO.1 - 2; primers for molecular marker RM5757, SEQ ID NO.3 - 4; primers for molecular marker H12, SEQ ID NO.5 - 6; primers for molecular marker H14, SEQ ID NO.7 - 8; primers for molecular marker 31 - 1, SEQ ID NO.9 - 10; primers for molecular marker 32 - 5, SEQ ID NO.11 - 12; primers for molecular marker H99, SEQ ID NO.13 - 14.

[0008] The third aspect of the present invention provides the application of the above - mentioned molecular marker primer composition in screening rice materials resistant to brown planthopper.

[0009] The fourth aspect of the present invention provides a kit for screening rice materials resistant to brown planthopper, and the kit contains the above - mentioned molecular marker primer composition.

[0010] The fifth aspect of the present invention provides the application of the above - mentioned kit in screening rice materials resistant to brown planthopper.

[0011] The advantages of the present invention are as follows: (1) The four double - gene pyramiding families carrying different insect - resistant genes obtained in the present invention are resistant or highly resistant to 7 different brown planthopper biotypes, and still remain resistant or highly resistant for a long time after insect inoculation, truly achieving durable broad - spectrum resistance to brown planthopper. Since the four gene combinations carry different genes from each other, it effectively ensures gene diversity, avoids the problem that a single gene is prone to failure, and provides the possibility for achieving durable broad - spectrum resistance to brown planthopper.

[0012] (2) The present invention can improve the efficiency of assisted selection in conventional breeding and save costs. By detecting the brown planthopper resistance gene loci in rice plants through molecular markers, resistant single plants can be quickly identified only at the seedling stage, and susceptible single plants can be eliminated in a timely manner without insect inoculation identification. This not only saves production costs, but also improves the selection efficiency of resistant materials and shortens the breeding cycle of rice varieties.

[0013] (3) By using the method of crossing single - gene near - isogenic lines with each other, the present invention can achieve the rapid pyramiding of multiple resistance genes under the same genetic background, reduce the detection steps of linkage drag and genetic background recovery rate, and improve the selection efficiency of pyramiding breeding. Description of the Drawings

[0014] Figure 1: Molecular marker gel images of homozygous resistant near-isogenic lines and F7 generations carrying resistance genes. H14, H99, H939, RM5757, 32-5, 31-1, and H12 are molecular markers, and resistance genes such as BPH14 and BPH33 in parentheses are resistance genes linked or co-segregating with these markers; 14, 33, 3, etc. respectively represent homozygous resistant near-isogenic lines carrying single resistance genes BPH14, BPH33, BPH3, etc., and 14 / / 33, 14 / / 3, etc. respectively represent F7 generations aggregating BPH14 and BPH33 genes, F7 generations aggregating BPH14 and BPH3 genes. Donor and 9311(ck) are the insect-resistant parent (donor) and the insect-susceptible parent (recipient) respectively; marker is the DNA size gradient indicator band, and the bands indicate 100bp, 200bp, 300bp, and 400bp from small to large.

[0015] Figure 2 : Genetic background analysis of F7 generations. The green bars represent all genomic fragment information of the recurrent parent 9311, the red bars represent genomic fragment information of the exogenous parent (other than 9311), and the positions indicated by the blue long arrows are the positions of the brown planthopper resistance genes BPH14, BPH33, BPH3, BPH6, BPH32, BPH31, and BPH9 on the chromosomes. The blue short arrows indicate the distribution of 48 pairs of SSR markers required for the national standard variety purity identification on the chromosomes.

[0016] Figure 3 : Plant type diagrams of homozygous resistant near-isogenic lines and F7 generations at the flowering stage in the field. 14, 33, 3, etc. respectively represent homozygous resistant near-isogenic lines carrying single resistance genes BPH14, BPH33, BPH3, etc., and 14 / 33, 14 / 3, etc. respectively represent F7 generations aggregating BPH14 and BPH33 genes, F7 generations aggregating BPH14 and BPH3 genes. Scale: 10 cm.

[0017] Figure 4 : Durable resistance of homozygous resistant near-isogenic lines and F7 generations. D10, D13, D16, D19, and D26 respectively represent the seedling stage resistance at 10 days, 13 days, 16 days, 19 days, and 26 days after insect inoculation; 14, 33, 3, etc. respectively represent homozygous resistant near-isogenic lines carrying single resistance genes BPH14, BPH33, BPH3, etc., and 14 / 33, 14 / 3, etc. respectively represent F7 generations aggregating BPH14 and BPH33 genes, F7 generations aggregating BPH14 and BPH3 genes. Scale: 10 cm. Detailed implementation manners

[0018] To make the content of the present invention easier to understand, the technical solutions of the present invention are further described below in conjunction with specific implementation manners, but the present invention is not limited thereto.

[0019] Seven homozygous resistant near-isogenic lines (NILs) under the background of indica rice 9311 used in the following examples, namely BPH3-NIL, BPH6-NIL, BPH9-NIL, BPH14-NIL, BPH31-NIL, BPH32-NIL, and BPH33-NIL, were obtained by using indica rice 9311 as the recurrent parent and introgressing the individual insect-resistant genes BPH3, BPH6, BPH9, BPH14, BPH31, BPH32, and BPH33 into 9311 through hybridization and backcrossing, and their construction methods have all been disclosed in the prior art. Among them, BPH3 is a gene cluster composed of three genes with gene numbers LOC_Os04g12540, LOC_Os04g12560, and LOC_Os04g12580; the gene number of BPH6 is LOC_Os04g35210; BPH9 is a gene cluster composed of two genes with gene numbers LOC_Os12g37280 and LOC_Os12g37290; the gene number of BPH14 is LOC_Os03g63150; the gene number of BPH31 is LOC_Os11g29150; the gene number of BPH32 is LOC_Os06g03240; the gene number of BPH33 is LOC_Os04g02520. For the construction methods of BPH3-NIL and BPH32-NIL, see: He et al. 2020. Molecular Breeding 40(10):95. DOI: 10.1007 / s11032-020-01175-z; for the construction method of BPH6-NIL, see: Xiao et al. 2016. Scientific Reports 6:38159. DOI: 10.1038 / srep38159; for the construction method of BPH9-NIL, see: Wang et al. 2017. Rice 10:51. DOI: 10.1186 / s12284-017-0194-x; for the construction method of BPH14-NIL, see: Hu et al. 2013. Pest Management Science 69:802-808. DOI: 10.1002 / ps.3437.; for the construction method of BPH31-NIL, see: Zhou Mingsong. Construction and Resistance Evaluation of Brown Planthopper Resistance Gene Pyramiding Lines in Rice and Fine Mapping of Bph31 [D]. Huazhong Agricultural University; for the construction method of BPH33-NIL, see: Hu et al. Rice 11:55. DOI: 10.1186 / s12284-018-0249-7.The donor of BPH3 is rice Rathu Heenati (RH), the donor of BPH6 is rice Swarnalata, the donor of BPH9 is rice Pokkali, the donor of BPH14 is rice B5, the donor of BPH31 is rice AUS69, the donor of BPH32 is rice Ptb33, and the donor of BPH33 is rice KOLAYAL.

[0020] The molecular marker information used in the MAS molecular marker-assisted breeding technology in the following examples is shown in Table 1.

[0021] Table 1 Molecular marker sequence information used in MAS The band size of the PCR amplification product of the primer of molecular marker H939 is as follows: if a single band of 132 bp appears, it means that the tested rice is a homozygous genotype of the BPH3 resistance allele (i.e., homozygous resistant genotype); if a single band of 142 bp appears, it means that the tested rice does not carry the BPH3 resistance allele (i.e., homozygous sensitive genotype); if there are two bands of 132 bp and 142 bp, it means that the tested rice is a heterozygous genotype of the BPH3 allele (i.e., heterozygous resistant genotype).

[0022] The band size of the PCR amplification product of the primer of molecular marker RM5757 is as follows: if a single band of 136 bp appears, it means that the tested rice is a homozygous genotype of the BPH6 resistance allele (i.e., homozygous resistant genotype); if a single band of 125 bp appears, it means that the tested rice does not carry the BPH6 resistance allele (i.e., homozygous sensitive genotype); if there are two bands of 136 bp and 125 bp, it means that the tested rice is a heterozygous genotype of Bph6 (i.e., heterozygous resistant genotype).

[0023] The band size of the PCR amplification product of the primer of molecular marker H12 is as follows: if a single band of 129 bp appears, it means that the tested rice is a homozygous genotype of the BPH9 resistance allele (i.e., homozygous resistant genotype); if a single band of 123 bp appears, it means that the tested rice does not carry the BPH9 resistance allele (i.e., homozygous sensitive genotype); if there are two bands of 129 bp and 123 bp, it means that the tested rice is a heterozygous genotype of Bph9 (i.e., heterozygous resistant genotype).

[0024] The band size of the PCR amplification product of the primer of molecular marker H14 is as follows: If a single band of 90 bp appears, it indicates that the tested rice is of the homozygous genotype of the BPH14 resistance allele (i.e., the homozygous resistant genotype); if a single band of 99 bp appears, it indicates that the tested rice does not carry the BPH14 resistance allele (i.e., the homozygous sensitive genotype); if there are two bands of 90 bp and 99 bp, it indicates that the tested rice is of the heterozygous genotype of Bph14 (i.e., the heterozygous resistant genotype).

[0025] The band size of the PCR amplification product of the primer of molecular marker 31-1 is as follows: If a single band of 198 bp appears, it indicates that the tested rice is of the homozygous genotype of the BPH31 resistance allele (i.e., the homozygous resistant genotype); if a single band of 163 bp appears, it indicates that the tested rice does not carry the BPH31 resistance allele (i.e., the homozygous sensitive genotype); if there are two bands of 198 bp and 163 bp, it indicates that the tested rice is of the heterozygous genotype of BPH31 (i.e., the heterozygous resistant genotype).

[0026] The band size of the PCR amplification product of the primer of molecular marker 32-5 is as follows: If a single band of 176 bp appears, it indicates that the tested rice is of the homozygous genotype of the BPH32 resistance allele (i.e., the homozygous resistant genotype); if a single band of 168 bp appears, it indicates that the tested rice does not carry the BPH32 resistance allele (i.e., the homozygous sensitive genotype); if there are two bands of 176 bp and 168 bp, it indicates that the tested rice is of the heterozygous genotype of BPH32 (i.e., the heterozygous resistant genotype).

[0027] The band size of the PCR amplification product of the primer of molecular marker H99 is as follows: If a single band of 96 bp appears, it indicates that the tested rice is of the homozygous genotype of the BPH33 resistance allele (i.e., the homozygous resistant genotype); if a single band of 102 bp appears, it indicates that the tested rice does not carry the BPH33 resistance allele (i.e., the homozygous sensitive genotype); if there are two bands of 96 bp and 102 bp, it indicates that the tested rice is of the heterozygous genotype of BPH33 (i.e., the heterozygous resistant genotype).

[0028] Example 1: (1) Using the homozygous resistant near-isogenic lines BPH3-NIL, BPH6-NIL, BPH9-NIL, BPH14-NIL, BPH31-NIL, BPH32-NIL, and BPH33-NIL each carrying a single insect-resistant gene as the basic materials, pairwise crosses and complete diallel crosses were carried out (i.e., BPH3-NIL was crossed with BPH6-NIL, BPH9-NIL, BPH14-NIL, BPH31-NIL, BPH32-NIL, and BPH33-NIL respectively, BPH6-NIL was crossed with BPH9-NIL, BPH14-NIL, BPH31-NIL, BPH32-NIL, and BPH33-NIL respectively, BPH9-NIL was crossed with BPH14-NIL, BPH31-NIL, BPH32-NIL, and BPH33-NIL respectively, BPH14-NIL was crossed with BPH31-NIL, BPH32-NIL, and BPH33-NIL respectively, BPH31-NIL was crossed with BPH32-NIL and BPH33-NIL respectively, and BPH32-NIL was crossed with BPH33-NIL), obtaining the F1 generations of 21 hybrid combinations, and ensuring that at least 20 hybrid seeds were obtained for each hybrid combination.

[0029] (2) After planting the F1 generations by plant lines, PCR detection was carried out using the MAS molecular marker-assisted breeding technology to screen out the F1 single plants with heterozygous resistant genotypes for the corresponding insect-resistant target genes, and the selfed seeds F2 were harvested after bagging.

[0030] (3) After planting the F2 generations by plant lines, PCR detection was carried out using the MAS molecular marker-assisted breeding technology to screen out the F2 single plants with homozygous resistant genotypes for the corresponding insect-resistant target genes, and the selfed seeds F3 were harvested after bagging.

[0031] (4) After planting the F3 generations by plant lines, selection was carried out according to the field agronomic traits in combination with the genetic background of the rice gene chip GSR40K, and the F3 single plants with no significant differences (P>0.05) in the agronomic traits (plant height, heading date, effective panicle number, filled grain number per panicle, 1000-grain weight, and yield per plant) from rice 9311 and a genetic background similarity of more than 95% were screened out, and the selfed seeds F4 were harvested after bagging.

[0032] (5) After planting the F4 generations by plant lines, selfing was carried out continuously for three generations to obtain the F7 generations, which were the 21 double-gene homozygous pyramiding families. Using the MAS molecular marker-assisted breeding technology to carry out PCR detection on the F7 generations, it was found that their corresponding insect-resistant target genes were all homozygous resistant genotypes ( Figure 1)。Six double-gene homozygous pyramiding families (BPH14 / BPH33, BPH14 / BPH32, BPH32 / BPH33, BPH3 / BPH6, BPH3 / BPH31, BPH3 / BPH9) were selected from the F7 generation, and rice 9311 was used as a control for rice gene chip GSR40K analysis. The results showed that the genetic background similarity between these six double-gene homozygous pyramiding families and the control rice 9311 was 95.45% - 98.46%, and there was only a small introduction of exogenous fragments linked to the target genes when the corresponding insect-resistant target genes all existed. Figure 2 )。In addition, the plant types of the F7 generation in the field were photographed, and the results showed that the plant types of most pyramiding families were very close to the recurrent parent rice 9311. Figure 3 )。

[0033] (6) The F7 generation and the recurrent parent rice 9311 were identified for their resistance to brown planthoppers at the seedling stage in the greenhouse. The identification method can be referred to: Huang et al. 2001. Theoretical and Applied Genetics 102: 929 - 934. https: / / doi.org / 10.1007 / s001220000455. The susceptible control rice 9311 and the materials to be identified were sown at the same time. 12 plump seeds of each material were sown in the same black plastic seedling box with an open bottom (diameter 7.2 cm, bottom diameter 5 cm, height 8 cm), and the seedling box was filled with soil suitable for growing rice. When the plants grew to the three-leaf stage, 9 seedlings with consistent growth were retained in each row, and the seedling tray box was placed inside a 200-mesh screen. Second-instar nymphs of brown planthoppers were introduced at a rate of 8 - 10 per plant. When more than 90% of the plants of the susceptible control died, the resistance level could be evaluated according to the damage degree of each plant, and the resistance score value of each material was obtained through weighted average calculation. When the dead seedling rate of 9311 reached 90%, resistance investigations and photography were carried out every 3 - 5 days. Finally, 14 double-gene homozygous pyramiding families were screened from the F7 generation, namely BPH14 / BPH33, BPH6 / BPH14, BPH9 / BPH14, BPH3 / BPH33, BPH6 / BPH33, BPH31 / BPH33, BPH32 / BPH33, BPH9 / BPH33, BPH3 / BPH6, BPH6 / BPH9, BPH6 / BPH31, BPH6 / BPH32, BPH32 / BPH31, and BPH9 / BPH31, which showed resistance or high resistance at all investigation periods, that is, they had durable resistance to brown planthoppers. Figure 4 )。

[0034] (7) Fourteen double-gene homozygous pyramiding families screened from the F7 generation in (6) were respectively introduced into 7 different populations of Nilaparvata lugens virulent types, and their broad-spectrum resistance was evaluated according to their resistance levels to different populations. Finally, 4 double-gene homozygous pyramiding families were screened from the F7 generation, namely BPH6 / BPH33, BPH14 / BPH33, BPH31 / BPH33, and BPH32 / BPH33, which showed resistance or high resistance to all 7 different populations of Nilaparvata lugens virulent types (Table 2). That is, these 4 double-gene pyramiding F7 are the novel rice materials with durable broad-spectrum resistance to Nilaparvata lugens referred to in the present invention.

[0035] Table 2 Seedling resistance of double-gene pyramiding lines with durable resistance to 7 Nilaparvata lugens populations Note: HR: High resistance; R: Resistance; MR: Moderate resistance; MS: Moderate susceptibility; S: Susceptibility; HS: High susceptibility. The families marked in bold showed resistance or high resistance to 7 Nilaparvata lugens populations.

[0036] The above are only the preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope covered by the present invention.

Claims

1. A method for creating a long-lasting and broad-spectrum brown planthopper-resistant rice material by multi-gene polymerization, characterized in that: The following steps are involved: (1) A homozygous resistant near-isogenic line carrying a single insect resistance gene in the rice 9311 background BPH3 -NIL, BPH6 -NIL, BPH9 -NIL, BPH14 -NIL, BPH31 -NIL, BPH32 -NIL and BPH33 -NIL is used as the basic material, and two-by-two hybridization and complete diallel hybridization are used to obtain the F1 generation of 21 hybrid combinations; (2) After the F1 generation is planted, the MAS molecular marker-assisted breeding technology is used to screen out the F1 plants with heterozygous resistance genotypes for the corresponding insect resistance target genes, and the plants are bagged and harvested from the cross F2; (3) After the F2 generation is planted, the MAS molecular marker-assisted breeding technology is used to screen out the F2 plants with homozygous resistance genotypes for the corresponding insect resistance target genes, and the self-cross F3 is collected by bagging; (4) After the F3 generation was planted, the F3 plants with similar agronomic traits to the indica rice 9311 and a genetic background similarity of more than 95% were selected based on the field agronomic traits combined with the genetic background selection of the rice gene chip, and the F4 plants were harvested in bags; (5) After the F4 generation was planted, it was self-pollinated for three consecutive generations to obtain the F7 generation; among them, the aggregate BPH6 and BPH33 F7 generation, polymer BPH14 and BPH33 F7 generation, polymer BPH31 and BPH33 F7 generation, polymer BPH32 and BPH33 The F7 generation is a rice material with long-lasting and broad-spectrum resistance to brown planthoppers.

2. The method according to claim 1, characterized in that: BPH3 It is from gene number LOC_Os04g12540 、 LOC_Os04g12560 、 A gene cluster consisting of three genes in LOC_Os04g12580; BPH6 The gene number is LOC_Os04g35210; BPH9 It is caused by gene number LOC_Os12g37280 、 A gene cluster consisting of 2 genes in LOC_Os12g37290; BPH14 The gene number is LOC_Os03g63150; BPH31 The gene number is LOC_Os11g29150; BPH32 The gene number is LOC_Os06g03240; BPH33 The gene number is LOC_Os04g02520.

3. The method according to claim 1, characterized in that: BPH3 The molecular marker is H939, and the primers of the molecular marker H939 are SEQ ID NO.1~2; BPH6 The molecular marker is RM5757, and the primers of the molecular marker RM5757 are SEQ ID NO.3~4; BPH9 The molecular marker is H12, and the primers of the molecular marker H12 are SEQ ID NO.5~6; BPH14 The molecular marker is H14, and the primers of the molecular marker H14 are SEQ ID NO.7-8; BPH31 The molecular marker is 31-1, and the primers of the molecular marker 31-1 are SEQ ID NO.9-10; BPH32 The molecular marker is 32-5, and the primers of the molecular marker 32-5 are SEQ ID NO.11~12; BPH33 The molecular marker is H99, and the primers of the molecular marker H99 are SEQ ID NO.13~14.

4. A molecular marker primer composition, characterized in that: The primer composition includes: primers for molecular marker H939, SEQ ID NOs. 1 to 2; primers for molecular marker RM5757, SEQ ID NOs. 3 to 4; primers for molecular marker H12, SEQ ID NOs. 5 to 6; primers for molecular marker H14, SEQ ID NOs. 7 to 8; primers for molecular marker 31-1, SEQ ID NOs. 9 to 10; primers for molecular marker 32-5, SEQ ID NOs. 11 to 12; and primers for molecular marker H99, SEQ ID NOs. 13 to 14.

5. Use of the molecular marker primer composition according to claim 4 in creating rice materials resistant to brown planthoppers.

6. A kit for creating rice materials resistant to brown planthoppers, characterized in that: The kit comprises the molecular marker primer composition according to claim 4.

7. Use of the kit according to claim 6 in creating rice materials resistant to brown planthoppers.

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