CN113796305A - Molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive planting and breeding in rice fields - Google Patents
Molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive planting and breeding in rice fields Download PDFInfo
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Abstract
The invention discloses a molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive planting and breeding in rice fields, which comprises the following steps: step 1: selecting a female parent and a male parent to be hybridized and matched to obtain first-generation seeds; step 2: planting F1After the plants are mature, all the single plants are harvested together; and step 3: planting F2，F2Molecular marker assistance of rice dark endosperm mutant gene, rice blast resistance gene, flooding resistance and sheath blight major resistance locus is carried out at the beginning of generationSelecting; and 4, step 4: f is to be2The generation contains WxmpThe gene homozygote and the homozygote and heterozygote individuals of the Pit, Sub1 and qSB-9 genes are respectively bred into F3A cell; and 5: planting F4The community has good lodging resistance and high yield, is consistent in performance and is resistant to sheath blight and rice blast; step 6: planting F5In tillering stage, the plots were tested for 7d and 14d flooding tolerance. The invention combines Pit, Sub1, qSB-9 and WxmpThe genes are integrated, and a new rice variety Nanjing 5718 with good resistance to rice blast and sheath blight, good taste quality and flooding resistance is bred.
Description
Technical Field
The invention relates to the technical field of rice breeding, in particular to a molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive rice field planting.
Background
The comprehensive planting and breeding of the rice field refers to the ecological cycle agricultural development mode which has multiple functions of grain stabilization, fishery promotion, quality improvement, efficiency improvement, ecology, environmental protection and the like and can realize stable rice yield, increase of aquatic product yield, increase of economic benefit and remarkable reduction of pesticide and fertilizer application amount through the engineering transformation of the rice field, the construction of a rice-fishery symbiotic rotation interaction promotion system and the large-scale development, the industrialized operation, the standardized production and the branded operation.
In recent years, with the reformation of agricultural supply sides, the comprehensive planting and breeding technology development of rice fields is promoted by the grabbing opportunity of agricultural departments at all levels in China. The comprehensive planting and breeding of the rice field is a production mode organically combining the planting industry and the breeding industry, and is an important technical measure for developing ecological agriculture and improving the comprehensive benefit of the rice field. The technical mode of comprehensive planting and breeding of rice and fisheries is popularized according to local conditions, the development requirement of ecological conservation and pollution reduction can be met, the agricultural efficiency increase and the income increase of farmers can be accelerated, and the cultivation of the rice variety special for comprehensive planting and breeding of the rice field is an important way for optimizing an agricultural structure, developing country characteristic industries and green agriculture.
Since the beginning of the 21 st century, after the whole genome sequencing of indica and japonica subspecies of rice is completed, the rice turns to functional genomics research from QTL positioning, and deeply excavates the functions of genes, explores the regulation and control network and genetic mechanism among genes, and lays a solid foundation for further and more effectively developing rice molecular breeding. In recent years, the rapid development of genome re-sequencing technology has led to the rapid development of rice genome, transcriptome, proteome, and even epigenomics, and rice molecular breeding has been achieved.
The method for rice gene location and molecular breeding is provided in the existing patent CN2017108223887, and the method for high-grade mapping population is combined on the basis of the low-grade mapping population, so that the time is saved, and the breeding efficiency is improved; the gene can be excavated for the positioning of new genes, and the molecular assisted breeding can be carried out for the cloned genes; a high-efficiency breeding method of rice with double resistance and double functions is provided in the prior patent CN2018106489077, two rice varieties with double functions and two rice varieties with disease resistance are taken as materials, and a new rice variety integrating double functions and double resistance into a whole can be obtained after hybridization and purification breeding.
At present, in the process of transferring agriculture to a mode and adjusting a structure, the comprehensive planting and breeding of the rice field is highly valued as an ecological agriculture and green agriculture production mode with high efficiency in output, resource conservation and environmental friendliness. Under the vigorous promotion of the Ministry of agriculture, under the active exploration and practice of governments, competent departments, scientific personnel and vast farmers in various regions, the comprehensive planting and development of the rice field achieves remarkable effect.
In the comprehensive planting and breeding composite ecological system of the rice field, rice and special aquatic products (fish, shrimp, crab and the like) are taken as centers. Because the use of pesticides can cause damage to special aquatic products, a high-quality rice variety with strong pest resistance and high lodging resistance needs to be selected. But the breeding of disease-resistant lodging-resistant high-quality high-yield rice varieties is still a difficult problem in the current rice field comprehensive planting and breeding process.
Disclosure of Invention
1. Technical problem to be solved
The invention aims to solve the problem of how to select and breed a high-quality disease-resistant flooding-resistant rice variety in the prior art, and provides a molecular breeding method for cultivating the high-quality rice variety suitable for comprehensive rice field breeding.
2. Technical scheme
In order to achieve the purpose, the invention adopts the following technical scheme:
a molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive planting and breeding in rice fields comprises the following steps:
step 1: selecting Jiangsu high-yield flood-resistant japonica rice variety salted japonica rice 608 with rice blast disease-resistant genotype PitPi and flood-resistant genotype Sub1Sub1 as female parent and gene with dark endosperm mutation with genotype WxmpWxmpThe high-quality japonica rice new line Ning 7022 containing the main effect QTLqSB-9 of the sheath blight resistance is taken as a male parent for hybridization and matching to obtain first-generation seeds;
step 2: planting F1After the plants are mature, all the single plants are harvested together;
and step 3: planting F2，F2Molecular marker-assisted selection of rice dark endosperm mutant gene, rice blast resistance gene, flooding-resistant and sheath blight major resistance locus is carried out at the beginning of generation, extraction of DNA of each individual plant in seedling stage adopts a CTAB method, genomic DNA of rice variety is amplified and observed after ethidium bromide staining, and Wx with two characteristic bands of 439bp and 292bp is selectedmpThe gene homozygote is used as a single plant for selectionA single plant containing a pure integration of the Pit gene with a 733bp characteristic band is reserved; selecting a single plant containing the Sub1 gene homozygote with a 586bp characteristic band for later use; selecting qSB-9 gene homozygote single plants with 253bp, 312bp and 335bp characteristic bands for later use;
and 4, step 4: f is to be2The generation contains WxmpThe gene homozygote and the homozygote and heterozygote individuals of the Pit, Sub1 and qSB-9 genes are respectively bred into F3The method comprises the following steps of (1) continuously detecting the 3-grade banded sclerotial blight resistance and rice blast immune or high-resistance cells by using molecular markers, and selecting the cells of which all single plants have characteristic bands of 439bp, 292bp, 733bp, 586bp, 253bp, 312bp and 335bp at the same time;
and 5: planting F4The community has good lodging resistance and high yield, is consistent in performance and is resistant to sheath blight and rice blast;
step 6: planting F5In tillering stage, carrying out 7d and 14d top-submerged flood-resistant tests on the small areas, selecting flood-resistant small areas capable of quickly recovering normal growth after flooding, and then carrying out Wx on the selected small areasmpThe Pit, Sub1 and qSB-9 genes are subjected to molecular verification to prevent target genes from being lost. Finally, in F6The rice strain with good taste, rice blast resistance, sheath blight resistance and flooding resistance is obtained.
Preferably, the japonica rice 608, the japonica rice variety resisting rice blast and sheath blight in Jiangsu in the step 1 and salt urban salt in Jiangsu province are bred by the agricultural department. The genotype at the Wx site is WxbwxbRice blast genotype PitPit, flood-resistant genotype Sub1Sub 1. The growth period is about 157 days, the plant height is about 102cm, the number of seeds per spike is 128-130, the seed setting rate is 94%, and the thousand seed weight is 27-28 g.
Preferably, step 1, zhongning 7022, Jiangsu high-quality and high-yield japonica rice varieties and grain crop research institute of agricultural science institute of Jiangsu province are bred, and the genotype at the Wx site is WxmpWxmpContains the major resistance QTLqSB-9 of the sheath blight disease. The variety has a full growth period of 156 days, a plant height of 97-100 cm, 8-10 effective spikes per plant, 115-125 seeds per spike, a setting rate of 92-93% and a thousand seed weight of 26-27 g.
Preferably, the dark endosperm mutation of rice in the step 3Gene WxmpHas two characteristic bands of 439bp and 292 bp.
Preferably, in the molecular detection of the rice blast resistance gene Pit in the step 3, the primers used comprise: a forward primer: 5'-ATGATAACCTCATAATCAATAAGT-3' and reverse primer: 5'-GTTGGAGCTACGGTTGTTCAG-3', respectively; the primers used for detecting the flooding-resistant gene Sub1 comprise: a forward primer: 5'-AGCAGGTTATAAGCTAGCTAT-3' and reverse primer: 5 '-CTACCAACAAGTTCATCAAAS-3'; the primers adopted for detecting the main effect QTLqSB-9 of the banded sclerotial blight comprise: y84-2 forward primer: 5'-AAAGGTTGCGAGGAGATTAGAGT-3' and reverse primer: 5'-TAGGGGTTGGTTTCTGGTTGTAG-3', respectively; y90.2 forward primer: 5'-GGGGATTAAATACGAGCAT-3' and reverse primer: 5'-TTTCTTAGGTCCCATTCTTC-3', respectively; y93.5 forward primer: 5'-CTGTTCTTCTCCTGCGTTCT-3' and reverse primer: 5'-ATGTCCTCGTGCTTCTGC-3' are provided.
Preferably, the step 4 contains Wx at the same timempSelecting single plants with plant height of 90-100 cm, 8-10 ears per plant and 140-150 grains per ear in the homozygote cell of the Pit, Sub1 and qSB-9 genes for addition generation stabilization, and performing F6The new rice strain Nanjing 5718 (field plot number L26) with stable and consistent agronomic characters, excellent taste, rice blast resistance and sheath blight resistance is obtained in the generation.
Preferably, the cells immunized or highly resistant to rice blast in the step 4 refer to cells having a rice blast infection rate of less than 5%. The anti-sheath blight cell is a cell with the disease grade of 3 and the lesion spots spread to the sheath of the 4 leaves.
Preferably, Wx in the step 4mpThe detection of the gene adopts a four-primer amplification hindered mutation system, and the name, the sequence and the expected size of an amplification fragment of a PCR amplification reaction primer are shown in Table 2.
Preferably, the 20 μ LPCR reaction system includes 2.0 μ L DNA (10 ng/. mu.L), 2.0 μ L Primer (4 pmol/. mu.L), 10 xBuffer (including MgCl)2) 2.0. mu.L, dNTP (2.5mmol/L)04. mu.L, Taq (5U/. mu.L) 0.5. mu.L and ddH2O13.1. mu.L, reaction program including pre-denaturation at 95 ℃ for 5 min; then denaturation at 95 deg.C for 30s, renaturation at 65 deg.C for 30s, extension at 72 deg.C for 1min, circulation for 35 times, extension at 72 deg.C for 7min, cooling at 10 deg.C for 10min, adding indicator into amplification product for use。
3. Advantageous effects
Compared with the prior art, the invention has the advantages that:
(1) in the invention, molecular markers of Pit, Sub1 genes and QTL qSB-9 and Wx are utilizedmpThe four-primer amplification hindered mutation system PCR detection technology of the gene detects target gene sites in different separation generations respectively, combines field multi-generation breeding, resistance identification, flooding resistance screening and kernel endosperm appearance identification, and successfully detects Pit, Sub1, qSB-9 and WxmpThe genes are integrated, the new rice variety Nanjing 5718 with good rice blast and sheath blight resistance, good taste quality and flooding resistance is bred, and the rice variety Nanjing 5718 is suitable for being planted in the North Huaihe area of Jiangsu province.
(2) According to the characteristics of rice variety special for comprehensive planting and breeding of rice field, such as sheath blight resistance, rice blast resistance, flooding resistance, strong stalks, big ears, developed root system and the like, the Jiangsu disease-resistant and high-yield japonica rice variety saline japonica rice 608 simultaneously carrying rice blast resistance gene Pit and flooding resistance gene Sub1 is used as a female parent, and the low-amylose-content gene Wx is carriedmpAnd the excellent taste japonica rice line Ning 7022 with the main effect of resisting the sheath blight QTL qSB-9 is used as a male parent to be configured with a hybridization combination for molecular marker polymerization breeding.
(3) The variety has obvious ideal plant type characteristics of proper plant height, moderate tillering, little ineffective tillering, thick and strong stems, developed root system and the like, has early mature period, resists rice blast and banded sclerotial blight, has obvious yield increase effect, and is a preferred variety for comprehensive planting and breeding of Jiangsu rice fields.
(4) In the invention, four sets of PCR detection systems are successfully applied to molecular marker-assisted selection, thereby not only providing a quick and efficient selection method for polygenic rice breeding, but also creating new germplasm resources for comprehensive rice field breeding.
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FIG. 1 is a breeding process of a molecular breeding method suitable for the comprehensive breeding of high-quality rice varieties in rice fields.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments.
Example 1:
a molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive planting and breeding in rice fields comprises the following steps:
step 1: selecting Jiangsu high-yield flood-resistant japonica rice variety salted japonica rice 608 with rice blast disease-resistant genotype PitPi and flood-resistant genotype Sub1Sub1 as female parent and gene with dark endosperm mutation with genotype WxmpWxmpThe high-quality japonica rice new line Ning 7022 containing the main effect QTLqSB-9 of resisting banded sclerotial blight is taken as a male parent for hybridization and matching to obtain first-generation seeds, namely japonica rice 608, Jiangsu japonica rice varieties resisting rice blast and banded sclerotial blight, which are bred by the agricultural department of salt city salt in Jiangsu province;
the genotype at the Wx site is WxbwxbRice blast genotype PitPit, flood-resistant genotype Sub1Sub 1. The growth period is about 157 days, the plant height is about 102cm, the number of seeds per ear is 128-130, the setting rate is 94%, the thousand seed weight is 27-28 g, Ning 7022, Jiangsu high-quality and high-yield japonica rice varieties, which are bred by food crop research institute of agriculture academy of sciences of Jiangsu province, and the genotype at Wx site is WxmpWxmpContains the major resistance QTLqSB-9 of the sheath blight disease. The variety has a full growth period of 156 days, a plant height of 97-100 cm, 8-10 effective spikes per plant, 115-125 seeds per spike, a setting rate of 92-93% and a thousand seed weight of 26-27 g;
step 2: planting F1After the plants are mature, all the single plants are harvested together;
and step 3: planting F2，F2Molecular marker-assisted selection of rice dark endosperm mutant gene, rice blast resistance gene, flooding-resistant and sheath blight major resistance locus is carried out at the beginning of generation, extraction of DNA of each individual plant in seedling stage adopts a CTAB method, genomic DNA of rice variety is amplified and observed after ethidium bromide staining, and Wx with two characteristic bands of 439bp and 292bp is selectedmpSelecting a single plant containing the Pit gene homozygote with a 733bp characteristic band for later use; selecting a single plant containing the Sub1 gene homozygote with a 586bp characteristic band for later use; selecting qSB-9 gene homozygote single plants with 253bp, 312bp and 335bp characteristic bands for standby,rice dark endosperm mutant gene WxmpHas two characteristic bands of 439bp and 292 bp;
the molecular detection of the rice blast resistance gene Pit adopts primers comprising: a forward primer: 5'-ATGATAACCTCATAATCAATAAGT-3' and reverse primer: 5'-GTTGGAGCTACGGTTGTTCAG-3', respectively; the primers used for detecting the flooding-resistant gene Sub1 comprise: a forward primer: 5'-AGCAGGTTATAAGCTAGCTAT-3' and reverse primer: 5 '-CTACCAACAAGTTCATCAAAS-3'; the primers adopted for detecting the main effect QTLqSB-9 of the banded sclerotial blight comprise: y84-2 forward primer: 5'-AAAGGTTGCGAGGAGATTAGAGT-3' and reverse primer: 5'-TAGGGGTTGGTTTCTGGTTGTAG-3', respectively; y90.2 forward primer: 5'-GGGGATTAAATACGAGCAT-3' and reverse primer: 5'-TTTCTTAGGTCCCATTCTTC-3', respectively; y93.5 forward primer: 5'-CTGTTCTTCTCCTGCGTTCT-3' and reverse primer: 5'-ATGTCCTCGTGCTTCTGC-3', respectively;
and 4, step 4: f is to be2The generation contains WxmpThe gene homozygote and the homozygote and heterozygote individuals of the Pit, Sub1 and qSB-9 genes are respectively bred into F3Detecting the 3-grade banded sclerotial blight resistance and the immune or high-resistance rice blast cell continuously by using the molecular marker, selecting the cells of which all single plants have 439bp, 292bp, 733bp, 586bp, 253bp, 312bp and 335bp characteristic bands simultaneously, and simultaneously containing WxmpSelecting single plants with plant height of 90-100 cm, 8-10 ears per plant and 140-150 grains per ear in the homozygote cell of the Pit, Sub1 and qSB-9 genes for addition generation stabilization, and performing F6The new rice strain Nanjing 5718 with stable and consistent agronomic characters, good taste, rice blast resistance and sheath blight resistance is obtained in the generation, and the immune or high-resistance rice blast cell refers to a cell with the rice blast infection rate of less than 5%. The anti-sheath blight cell is a cell with disease grade of 3 and the lesion is spread to the sheath of 4 leaves, WxmpThe detection of the gene adopts a four-Primer amplification hindered mutation system, the name, sequence and expected size of an amplification fragment of a PCR amplification reaction Primer are shown in Table 2, a 20 mu LPCR reaction system comprises 2.0 mu L of DNA (10 ng/mu L), 2.0 mu L of Primer (4 pmol/mu L) and 10 x Buffer (comprising MgCl)2) 2.0. mu.L, dNTP (2.5mmol/L) 0.4. mu.L, Taq (5U/. mu.L) 0.5. mu.L and ddH2O13.1. mu.L, reaction program including pre-denaturation at 95 ℃ for 5 min; then denatured at 95 ℃ for 30 ℃s, renaturation at 65 ℃ for 30s, extension at 72 ℃ for 1min, circulation for 35 times, extension at 72 ℃ for 7min, cooling at 10 ℃ for 10min, and adding an indicator into an amplification product for later use;
and 5: planting F4The community has good lodging resistance and high yield, is consistent in performance and is resistant to sheath blight and rice blast;
step 6: planting F5In tillering stage, carrying out 7d and 14d top-submerged flood-resistant tests on the small areas, selecting flood-resistant small areas capable of quickly recovering normal growth after flooding, and then carrying out Wx on the selected small areasmpThe Pit, Sub1 and qSB-9 genes are subjected to molecular marker verification to prevent target genes from being lost. Finally, in F6The Nanjing 5718 (field plot number L26) of the flooding-resistant rice strain with excellent taste, rice blast resistance, sheath blight resistance and rice seedling resistance is obtained.
In the present invention, Table 1 shows the names and sequences of primers used for detecting the Pit, Sub1, qSB-9 genes and the expected fragment lengths
Note: the double-sided polymorphic molecular markers Y84-2, Y90.2 and Y93.5 of QTLqSB-9.
In the present invention, Table 2 is used for the detection of WxmpPrimer name, sequence and expected fragment length of gene
Note: wxmpThe amplified products of different genotypes have banding patterns, the dominant homozygote is 439bp and 292bp, the recessive homozygote is 439bp and 200bp, and the heterozygote is 439bp, 292bp and 200 bp.
In the present invention, WxmpThe detection of the gene adopts a four-primer amplification hindered mutation system reported by Chen and the like. PCR amplification reaction primer name, sequence and amplification piece thereofThe expected sizes of the segments are shown in Table 2. A20. mu.L PCR reaction system included 2.0. mu.L of DNA (10 ng/. mu.L), 2.0. mu.L of Primer (4 pmol/. mu.L), 10 XBuffer (including MgCl)2) 2.0. mu.L, dNTP (2.5mmol/L) 0.4. mu.L, Taq (5U/. mu.L) 0.5. mu.L and ddH2O13.1. mu.L. The reaction procedure included pre-denaturation at 95 ℃ for 5 min; then, denaturation at 95 ℃ for 30s, renaturation at 65 ℃ for 30s, extension at 72 ℃ for 1min, circulation for 35 times, extension at 72 ℃ for 7min, cooling at 10 ℃ for 10min, and adding an indicator into an amplification product for later use.
In the invention, the salt japonica 608, Jiangsu japonica rice varieties resisting rice blast and banded sclerotial blight, and Jiangsu province salt city salt are bred by the agricultural department. The genotype at the Wx site is WxbwxbRice blast genotype PitPit, flood-resistant genotype Sub1Sub 1. The growth period is about 157 days, the plant height is about 102cm, the number of seeds per spike is 128-130, the seed setting rate is 94%, and the thousand seed weight is 27-28 g.
In the invention, Ning 7022, Jiangsu high-quality and high-yield japonica rice varieties are bred by grain crop research institute of agricultural science institute of Jiangsu province, and the genotype at Wx site is WxmpWxmpContains the major resistance QTLqSB-9 of the sheath blight disease. The variety has a full growth period of 156 days, a plant height of 97-100 cm, 8-10 effective spikes per plant, 115-125 seeds per spike, a setting rate of 92-93% and a thousand seed weight of 26-27 g.
In the present invention, Table 3 shows the comparison of flooding resistance
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (9)
1. A molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive planting and breeding in rice fields is characterized by comprising the following steps:
step 1: selecting Jiangsu high-yield flood-resistant japonica rice variety salted japonica rice 608 with rice blast disease-resistant genotype PitPi and flood-resistant genotype Sub1Sub1 as female parent and gene with dark endosperm mutation with genotype WxmpWxmpThe high-quality japonica rice new line Ning 7022 containing the main effect QTLqSB-9 of the sheath blight resistance is taken as a male parent for hybridization and matching to obtain first-generation seeds;
step 2: planting F1After the plants are mature, all the single plants are harvested together;
and step 3: planting F2，F2Molecular marker-assisted selection of rice dark endosperm mutant gene, rice blast resistance gene, flooding-resistant and sheath blight major resistance locus is carried out at the beginning of generation, extraction of DNA of each individual plant in seedling stage adopts a CTAB method, genomic DNA of rice variety is amplified and observed after ethidium bromide staining, and Wx with two characteristic bands of 439bp and 292bp is selectedmpSelecting a single plant containing the Pit gene homozygote with a 733bp characteristic band for later use; selecting a single plant containing the Sub1 gene homozygote with a 586bp characteristic band for later use; selecting qSB-9 gene homozygote single plants with 253bp, 312bp and 335bp characteristic bands for later use;
and 4, step 4: f is to be2The generation contains WxmpThe gene homozygote and the homozygote and heterozygote individuals of the Pit, Sub1 and qSB-9 genes are respectively bred into F3The method comprises the following steps of (1) continuously detecting the 3-grade banded sclerotial blight resistance and rice blast immune or high-resistance cells by using molecular markers, and selecting the cells of which all single plants have characteristic bands of 439bp, 292bp, 733bp, 586bp, 253bp, 312bp and 335bp at the same time;
and 5: planting F4The community has good lodging resistance and high yield, is consistent in performance and is resistant to sheath blight and rice blast;
step 6: planting F5In tillering stage, carrying out 7d and 14d top-submerged flood-resistant tests on the small areas, selecting flood-resistant small areas capable of quickly recovering normal growth after flooding, and then carrying out Wx on the selected small areasmpThe Pit, Sub1 and qSB-9 genes are subjected to molecular verification to prevent target genes from being lost, and finally,in F6The rice strain with good taste, rice blast resistance, sheath blight resistance and flooding resistance is obtained.
2. The molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive rice field cultivation according to claim 1, wherein the japonica rice with salt 608, the japonica rice with rice blast resistance and sheath blight resistance of Jiangsu, and the japonica rice with salt city of Jiangsu province are bred by the agricultural science, and the genotype at Wx site is WxbwxbThe rice blast genotype PitPoint, the flooding-resistant genotype Sub1Sub1, the growth period about 157 days, the plant height about 102cm, the number of seeds per spike 128-130, the seed setting rate 94% and the thousand seed weight 27-28 g.
3. The molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive rice field cultivation according to claim 1, wherein step 1, Zhongning 7022, Jiangsu high-quality and high-yield japonica rice varieties, grain crop research institute of agriculture academy of Jiangsu province, and the genotype at Wx site is WxmpWxmpThe bacterial wilt resistant strain contains QTLqSB-9 with the main effect resistance of the bacterial wilt, the full growth period of the strain is 156 days, the plant height is 97-100 cm, 8-10 effective spikes are provided for each plant, the number of seeds of each spike is 115-125, the seed setting rate is 92-93%, and the thousand seed weight is 26-27 g.
4. The molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive rice field cultivation according to claim 1, wherein the mutant gene Wx of dark endosperm of rice in the step 3 ismpHas two characteristic bands of 439bp and 292 bp.
5. The molecular breeding method for cultivating the high-quality rice varieties suitable for comprehensive rice field cultivation according to claim 1, wherein the molecular detection of the blast disease resistance gene Pit in the step 3 comprises the following primers: a forward primer: 5'-ATGATAACCTCATAATCAATAAGT-3' and reverse primer: 5'-GTTGGAGCTACGGTTGTTCAG-3', respectively; the primers used for detecting the flooding-resistant gene Sub1 comprise: a forward primer: 5'-AGCAGGTTATAAGCTAGCTAT-3' and reverse primer: 5 '-CTACCAACAAGTTCATCAAAS-3'; the primers adopted for detecting the main effect QTLqSB-9 of the banded sclerotial blight comprise: y84-2 forward primer: 5'-AAAGGTTGCGAGGAGATTAGAGT-3' and reverse primer: 5'-TAGGGGTTGGTTTCTGGTTGTAG-3', respectively; y90.2 forward primer: 5'-GGGGATTAAATACGAGCAT-3' and reverse primer: 5'-TTTCTTAGGTCCCATTCTTC-3', respectively; y93.5 forward primer: 5'-CTGTTCTTCTCCTGCGTTCT-3' and reverse primer: 5'-ATGTCCTCGTGCTTCTGC-3' are provided.
6. The molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive rice field cultivation according to claim 1, wherein step 4 simultaneously contains WxmpSelecting single plants with plant height of 90-100 cm, 8-10 ears per plant and 140-150 grains per ear in the homozygote cell of the Pit, Sub1 and qSB-9 genes for addition generation stabilization, and performing F6The new rice strain Nanjing 5718 (field plot number L26) with stable and consistent agronomic characters, excellent taste, rice blast resistance and sheath blight resistance is obtained in the generation.
7. The molecular breeding method for cultivating the high-quality rice varieties suitable for the comprehensive rice field planting of claim 1, wherein the cells immune or highly resistant to rice blast in the step 4 refer to cells with a rice blast infection rate of less than 5%, the cells resistant to sheath blight refer to cells with a disease grade of 3, and the disease spots extend to the upper part of the sheath of the 4 leaves.
8. The molecular breeding method for cultivating high-quality rice varieties suitable for comprehensive rice field cultivation according to claim 1, wherein Wx in the step 4mpThe detection of the gene adopts a four-primer amplification hindered mutation system, and the name, the sequence and the expected size of an amplification fragment of a PCR amplification reaction primer are shown in Table 2.
9. The molecular breeding method for cultivating high-quality rice cultivars suitable for rice field comprehensive cultivation according to claim 8, wherein the 20 μ LPCR reaction system comprises DNA (10ng/μ L)2.0 μ L, Primer (4pmol/μ L2).0 μ L, and 10 xBuffer (including MgCl22) mu.L, dNTP (2.5mmol/L0)4. mu.L, Taq (5U/. mu.L) 0.5. mu.L and ddH2O13.1. mu.L, reaction program including pre-denaturation at 95 ℃ for 5 min; then, denaturation at 95 ℃ for 30s, renaturation at 65 ℃ for 30s, extension at 72 ℃ for 1min, circulation for 35 times, extension at 72 ℃ for 7min, cooling at 10 ℃ for 10min, and adding an indicator into an amplification product for later use.
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