CN112931188A - Method for breeding new rice variety with wild rice genetic background - Google Patents
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Abstract
The invention relates to a method for breeding a new rice variety with a wild rice genetic background, which utilizes a rice genomics technology to rapidly and accurately breed the new rice variety with the wild rice genetic background, and more importantly, the inventor of the invention improves the screening condition in the breeding process, overcomes the technical difficulty and bottleneck of the hybrid breeding process of the wild rice and cultivated rice, can breed the new rice variety with the wild rice genetic background, effectively enlarges the genetic background of the cultivated rice, solves the serious problem of the homogenization of the existing variety, is favorable for creating novel germplasm resources, improves the yield of the hybrid rice and obtains breakthrough varieties.
Description
Technical Field
The invention relates to a method for breeding a new rice variety with a wild rice genetic background, in particular to a method for rapidly and accurately breeding a new rice variety with a wild rice genetic background by using a rice genomics technology, and belongs to the technical field of rice breeding.
Background
Rice is a main food crop in China, the global natural environment is increasingly worsened since the 21 st century, extreme weather frequently occurs, and new requirements are provided for breeding new varieties of rice.
Because the existing rice varieties have the problem of serious homogenization, if extreme natural climate disasters occur, destructive impact is brought to the high yield and the stable yield of the rice. Therefore, the method expands the genetic background of the parents, creates germplasm resources with rich genetic background, and is a main method for breeding new varieties of high-yield hybrid rice.
At present, breeders mainly adopt indica-japonica hybrid or local variety introduction into cultivated varieties to expand the genetic background of parents.
Wild rice, as a wild related species of cultivated rice, has many high-quality genes which are lost or not possessed by cultivated rice, such as related genes of disease and pest resistance, stress tolerance, high yield and the like, and is a natural genetic germplasm resource bank for further improving and expanding genetic backgrounds of cultivated rice parent varieties.
In the current wild rice-cultivated rice breeding practice, the wild rice is usually directly used as a donor, the cultivated rice is used as an acceptor, and a new germplasm resource with the wild rice genetic background is bred by a common hybridization and pedigree selection method.
However, wild rice and cultivated rice have a far relationship of parents and a large genetic background difference, so that progeny generated by the cross breeding of the wild rice and the cultivated rice has a plurality of defects, such as large phenotypic segregation difference, biased phenotype towards the wild rice, linkage drag, long segregation generation, difficult stability of genetic characters and the like.
How to overcome the technical difficulties and bottlenecks in the crossbreeding process of the wild rice and the cultivated rice and obtaining a new rice variety with the genetic background of the wild rice is a problem to be solved by technical personnel in the field.
Disclosure of Invention
In order to solve the above technical problems, the present invention provides, in one aspect, a method for breeding a new variety of rice with a genetic background of wild rice, wherein,
the method comprises the following steps of:
step 1): carrying out conventional sexual hybridization by taking a cultivated rice variety as a female parent and an AA type wild rice variety as a male parent to obtain a hybrid F1 generation;
step 2): planting the hybrid seeds obtained in the step 1) for generation F1; carrying out first backcross by taking the hybrid F1 generation as a female parent and the cultivated rice variety as a male parent to obtain BC1F1 generation seeds;
step 3): planting the BC1F1 generation seeds obtained in the step 2) to obtain a BC1F1 generation plant population, and taking leaves of the BC1F1 generation plant population at a seedling stage to perform whole genome sequencing with low coverage rate to obtain a genotype map of each BC1F1 generation plant;
screening suitable BC1F1 generation plants under the following conditions: the screened plants each contain wild rice genome segments in their genome, and the collection of different wild rice genome segments contained in these plants can cover 12 chromosomes of the wild rice genome; and, these plants, in addition to containing wild rice genomic fragments, all derive their genetic background from the oryza sativa genome;
step 4): carrying out second backcross by taking the proper plants of the BC1F1 generation screened out in the step 3) as female parents and the cultivated rice variety as male parents to obtain seeds of the BC2F1 generation;
or planting the plants of the BC1F1 generation selected in the step 3), and performing selfing to obtain seeds of the BC1F2 generation;
step 5): planting the BC2F1 generation seeds or BC1F2 generation seeds obtained in the step 4) to obtain respective plant populations, and taking leaves of the plant populations at a seedling stage to perform whole genome sequencing with low coverage rate to obtain a genotype map of each plant;
screening suitable plants of BC2F1 generation or BC1F2 generation under the following screening conditions: wild rice gene segments contained in the plant account for 5 to 20 percent of the genome background; and, except the wild rice gene contained, the genetic background is derived from the cultivated rice genome, and the screening criterion of the degree of purity is more than 60%; and, its agronomic traits more closely approximate those of the cultivar of rice;
planting the screened proper plants of the BC2F1 generation or BC1F2 generation, and performing selfing to obtain seeds of the respective next generation;
step 6): repeating at least 3 generations according to the mode in the step 5) until a stable new rice variety with wild rice genetic background is obtained.
Preferably, in the step 4), BC2F1 generation seeds are obtained; screening to obtain proper plants of the BC2F1 generation in the subsequent step 5), and performing selfing to obtain seeds of the BC2F2 generation; in the subsequent step 6), selfing was repeated for 3 generations to obtain stable seeds of BC2F5 generation with a wild rice genetic background.
More preferably, the AA type wild rice variety is "374"; the cultivated rice variety is 'WYG'.
Preferably, in the step 4), BC1F2 generation seeds are obtained; screening to obtain proper plants of the BC1F2 generation in the subsequent step 5), and performing selfing to obtain seeds of the BC1F3 generation; in the subsequent step 6), selfing was repeated for 2 generations to obtain stable seeds of BC1F5 generation with a wild rice genetic background.
More preferably, the AA type wild rice variety is W3105; the cultivated rice is Wushan silk seedling.
Preferably, during said steps 5) and 6), the lowest value of the selection criteria for the degree of homozygosity in said selection conditions is increased by 10% after each selfing generation.
Preferably, in the screening conditions in step 5), the selection criteria of the agronomic shape comprise: the plant height is 50-140 cm, the tillering number is more than 5, and the setting percentage is more than 60%.
Preferably, the whole genome sequencing with low coverage rate is carried out by using an Illumina Hiseq X10 sequencing platform and carrying out whole genome re-sequencing by using a 2X 150bp double-end sequencing method, wherein the sequencing depth of a parent is about 50X, and the sequencing depth of a progeny single plant is about 0.1X.
Preferably, in the step 5), the process of screening suitable plants further comprises performing combining ability measurement on plants meeting the screening conditions, and selecting plants with high combining ability. .
The invention provides a method for breeding a new rice variety with a wild rice genetic background, which utilizes a rice genomics technology to rapidly and accurately breed the new rice variety with the wild rice genetic background, and more importantly, the inventor of the invention improves the screening condition in the breeding process, overcomes the technical difficulty and bottleneck of the hybrid breeding process of the wild rice and cultivated rice, can breed the new rice variety with the wild rice genetic background, effectively enlarges the genetic background of the cultivated rice, solves the serious problem of the homogenization of the existing variety, is beneficial to creating novel germplasm resources, improves the yield of the hybrid rice and obtains breakthrough varieties.
Detailed Description
The present invention will be further described with reference to the following examples, but the present invention is not limited to these specific embodiments.
The explanation about the terms is as follows:
the term "cultivar of oryza sativa" means a rice species suitable for cultivation from wild rice through natural selection and artificial selection for a long period of time, for example, Huazhan, Huanghuazhan, Wushan rice seedlings and the like;
the term "AA type wild rice variety" means that the wild rice genome is an AA type genome.
The term "conventional sexual hybridization" refers to a technique in which male and female cells are combined by pollination to produce a hybrid.
The term "sequencing depth", the corresponding english term: sequencing Depth refers to the ratio of the total base number (bp) obtained by Sequencing to the Genome size (Genome), and is one of the indexes for evaluating the Sequencing quantity.
The term "low coverage whole genome sequencing", also known as "low depth whole genome sequencing". The genome sequencing depth refers to the ratio of the total base number obtained by sequencing to the size of the genome to be detected; in the protocol of the present application, the sequencing depth of the parents (i.e., cultivars of oryza sativa and AA type wild rice) is about 50 x and the sequencing depth of the individual progeny is about 0.1 x.
The term "genotype map" is drawn as a linear arrangement of the cultivated rice gene types or wild rice gene types on the chromosome.
The term "homozygosity" refers to the proportion of homozygous loci in an individual.
In one embodiment of the present invention, a method for breeding a new variety of rice with a genetic background of wild rice is provided, wherein,
the method comprises the following steps of:
step 1): carrying out conventional sexual hybridization by taking a cultivated rice variety as a female parent and an AA type wild rice variety as a male parent to obtain a hybrid F1 generation;
step 2): planting the hybrid seeds obtained in the step 1) for generation F1; carrying out first backcross by taking the hybrid F1 generation as a female parent and the cultivated rice variety as a male parent to obtain BC1F1 generation seeds;
step 3): planting the BC1F1 generation seeds obtained in the step 2) to obtain a BC1F1 generation plant population, and taking leaves of the BC1F1 generation plant population at a seedling stage to perform whole genome sequencing with low coverage rate to obtain a genotype map of each BC1F1 generation plant;
screening suitable BC1F1 generation plants under the following conditions: the screened plants each contain wild rice genome segments in their genome, and the collection of different wild rice genome segments contained in these plants can cover 12 chromosomes of the wild rice genome; and, these plants, in addition to containing wild rice genomic fragments, all derive their genetic background from the oryza sativa genome;
step 4): carrying out second backcross by taking the proper plants of the BC1F1 generation screened out in the step 3) as female parents and the cultivated rice variety as male parents to obtain seeds of the BC2F1 generation;
or planting the plants of the BC1F1 generation selected in the step 3), and performing selfing to obtain seeds of the BC1F2 generation;
step 5): planting the BC2F1 generation seeds or BC1F2 generation seeds obtained in the step 4) to obtain respective plant populations, and taking leaves of the plant populations at a seedling stage to perform whole genome sequencing with low coverage rate to obtain a genotype map of each plant;
screening suitable plants of BC2F1 generation or BC1F2 generation under the following screening conditions: wild rice gene segments contained in the plant account for 5 to 20 percent of the genome background; and, except the wild rice gene contained, the genetic background is derived from the cultivated rice genome, and the screening criterion of the degree of purity is more than 60%; and, its agronomic traits more closely approximate those of the cultivar of rice;
planting the screened proper plants of the BC2F1 generation or BC1F2 generation, and performing selfing to obtain seeds of the respective next generation;
step 6): repeating at least 3 generations according to the mode in the step 5) until a stable new rice variety with wild rice genetic background is obtained.
As explained in the background of the present application, those skilled in the art would like to be able to breed new varieties of rice with wild rice genetic background, but the practical methods in the prior art cannot quickly breed good varieties or germplasm resources, mainly because: wild rice and cultivated rice have far-distant parent relations and large genetic background difference, so that offspring produced by the cross breeding of the wild rice and the cultivated rice has a plurality of defects, such as large phenotype segregation difference, phenotype bias to the wild rice, linkage drag, long segregation generation, difficult stability of genetic characters and the like.
Therefore, the inventor of the present application hopes to improve the breeding method based on the prior art, overcome the technical difficulties and bottlenecks of the cross breeding process of the wild rice and the cultivated rice, and obtain a new rice variety or germplasm resource with the genetic background of the wild rice.
For this development direction, the inventors made 4 years of attempts in breeding practice, which involved extensive field planting and data collection, as well as extensive genome sequencing work and data statistics and analysis, and finally found, through the attempts of various schemes, that when the screening conditions in step 5) and step 6) were satisfied: wild rice gene segments contained in the plant account for 5 to 20 percent of the genome background; and, except the wild rice genome contained, the genetic background is derived from the cultivated rice genome, and the screening criterion of the degree of purity is 60% or more; and, the agronomic characters of the rice are closer to the cultivated rice variety, and when the conditions are the same, a stable new rice variety or germplasm resource with wild rice genetic background can be bred; particularly, in the processes of the step 5) and the step 6), after each selfing generation, the lowest value of the purity degree screening standard in the screening condition is improved by 10% until the purity degree reaches 100%, so that the operation process of breeding can be further simplified, and the breeding time is shortened.
In the screening conditions, the agronomic characters are closer to the cultivated rice varieties, and the technicians in the field generally judge and operate according to the accumulated experience and common knowledge; however, the two data, namely 5% -20% (the proportion of wild rice gene fragments contained in the plant to the genome background) and more than 60% (the degree of purity of the cultivated rice genome), are extremely critical, and are the crucial technical conditions for overcoming the technical difficulties and bottlenecks of the hybrid breeding process of the wild rice and the cultivated rice, which are summarized by a large number of attempts, data statistics and analysis by the inventor.
In addition, the breeding method of the invention utilizes the genotyping technology based on the whole genome sequencing of the rice to carry out genetic background selection, thereby shortening the improved generation of the wild rice-cultivated rice, breeding a new variety in 3-4 years and greatly accelerating the utilization efficiency of the wild rice; more importantly, the breeding method of the invention effectively expands the genetic background of the cultivated rice, solves the serious problem of homogenization of the existing variety, is beneficial to creating novel germplasm resources, improves the yield of hybrid rice and obtains breakthrough varieties.
Example 1 Process for Breeding New Rice variety
Step 1): carrying out conventional sexual hybridization by taking a cultivated rice variety (WYG with good comprehensive agronomic characters) as a female parent and an AA type wild rice variety (AA type common wild rice '374') as a male parent to obtain a hybrid F1 generation;
step 2): planting the hybrid seeds F1 obtained in the step 1); carrying out first backcross by taking the hybrid F1 generation as a female parent and the cultivated rice WYG as a male parent to obtain BC1F1 generation seeds;
step 3): planting the BC1F1 generation seeds obtained in the step 2) to obtain a BC1F1 generation plant population, and performing whole genome sequencing on leaves of the BC1F1 generation plant population at a seedling stage with low coverage rate to obtain a genotype map of each BC1F1 generation plant;
the specific procedures for low coverage whole genome sequencing and genotyping are as follows:
i) the cultivated rice "WYG", the wild rice "374" and the plant population of BC1F1 generation obtained above (146 BC1F1 individuals were obtained in this example) were numbered, the leaves of the corresponding plants were taken at the seedling stage, and the genomic DNA of each plant leaf was extracted using the new plant genomic DNA extraction kit N96 from Qiaqen;
ii) breaking the genome DNA, repairing with DNA polymerase, connecting with a linker, performing electrophoresis, recovering the fragment of 300-400bp, and establishing a PE400 sequencing library;
iii) carrying out whole genome re-sequencing by using an Illumina Hiseq X10 sequencing platform and a 2X 150bp double-end sequencing method, wherein the sequencing depth of a parent is about 50X, the sequencing depth of a progeny single plant is about 0.1X, and all sequencing work can be completed within about one week;
iv) obtaining the genotype map of the single filial generation plants by carrying out whole genome sequence alignment and genetic variation analysis and identification on the single filial generation plants (each BC1F1 generation plant) and parents ('WYG' and '374').
And (3) screening suitable BC1F1 generation plants according to a genotype map, wherein the screening conditions are as follows: the plant contains wild rice gene segments on each chromosome; besides the wild rice gene, the other genetic background is derived from the cultivated rice genome;
in the screening, although each plant contains a wild rice gene, plants having a low degree of heterozygosity and a high degree of homozygosity are selected as much as possible.
In this embodiment, some plants with better leaf morphology, which is closer to cultivated rice variety, can be further selected.
Specifically, in this example, 48 BC1F1 generation individuals satisfying the above requirements were selected.
Step 4): carrying out second backcross by taking proper plants of BC1F1 generation selected in the step 3) as female parents and 'WYG' as male parents to obtain seeds of BC2F1 generation;
step 5): planting the BC2F1 generation seeds obtained in the step 4) to obtain respective plant populations, and taking leaves of the plant populations at the seedling stage to perform whole genome sequencing with low coverage rate to obtain the genotype map of each plant;
screening suitable BC2F1 generation plants under the following conditions: wild rice gene segments contained in the plant account for 5 to 20 percent of the genome background; besides the wild rice genome, other genetic backgrounds are derived from the cultivated rice genome, and the degree of purity is more than 60%; and, its agronomic trait is closer to cultivated rice "WYG";
selection criteria for agronomic traits include: the plant height is 50-140 cm, the tillering number is more than 5, and the setting percentage is more than 60%; the variety closer to cultivated rice "WYG" was selected as much as possible.
In addition, the agronomic traits include the shatterability, fertility, and leaf morphology, and the selection of these traits can be made by those skilled in the art according to the ultimate goal and need of breeding.
In this embodiment, some plants with better leaf morphology, which is closer to cultivated rice variety, can be further selected.
Specifically, in this example, 15 plants of BC2F1 generation meeting the above requirements were finally screened.
Planting the screened proper plants of the BC2F1 generation, and performing selfing to obtain seeds of the respective BC2F2 generation;
regarding the way of selfing, bagging selfing is adopted in this embodiment.
Step 6): repeating at least 3 generations according to the mode in the step 5) until a stable new rice variety with wild rice genetic background is obtained.
Specifically, in this embodiment, in step 5) and step 6), after each selfing generation, the lowest value of the fitness screening criteria in the screening conditions is increased by 10% until 100% is reached.
Namely, planting seeds of BC2F2 generation to obtain a plant group, screening proper plants of BC2F2 generation according to the screening mode and the purity of the genome background of the cultivated rice is more than 70%, and selfing to obtain seeds of BC2F3 generation;
planting seeds of BC2F3 generation to obtain plant population, screening proper plants of BC2F3 generation according to the screening mode and the purity of the genome background of the cultivated rice is more than 80%, and selfing to obtain seeds of BC2F4 generation;
planting seeds of BC2F4 generation to obtain plant population, screening proper plants of BC2F4 generation according to the screening mode and the purity of the genome background of the cultivated rice is more than 90%, and selfing to obtain seeds of BC2F5 generation;
planting seeds of BC2F5 generation to obtain a plant population, and screening out proper plants of BC2F5 generation according to the screening mode and the purity of the genome background of the cultivated rice of 100 percent; that is, the wild rice gene segments contained in the screened plants account for 5 to 20 percent of the genome background; and, besides the wild rice gene contained, the genetic background is derived from the cultivated rice genome, and the degree of purity of the cultivated rice genome background is 100%, and the overall traits are stable. Specifically, in this example, 11 BC2F5 individuals were finally selected as new varieties of rice that are stable and have a wild rice background, and they were named as Y-1 to Y-11.
Example 2 Process for Breeding New Rice variety
Step 1): carrying out conventional sexual hybridization by taking a cultivated rice variety (a 'Wushan rice seedling' with good comprehensive agronomic characters) as a female parent and an AA type wild rice variety (an AA type common wild rice 'W3105') as a male parent to obtain a hybrid F1 generation;
step 2): planting the hybrid seeds F1 obtained in the step 1); carrying out first backcross by taking the hybrid F1 generation as a female parent and the cultivated rice Wushan Yangming as a male parent to obtain BC1F1 generation seeds;
step 3): planting the BC1F1 generation seeds obtained in the step 2) to obtain a BC1F1 generation plant population, and performing whole genome sequencing on leaves of the BC1F1 generation plant population at a seedling stage with low coverage rate to obtain a genotype map of each BC1F1 generation plant;
the specific procedures for low coverage whole genome sequencing and genotyping are as follows:
i) the cultivated rice "wushan silk seedling", wild rice "W3105" and the plant population of BC1F1 generation obtained above (230 BC1F1 individuals were obtained in this example) were numbered, and the leaf of the corresponding plant was taken at the seedling stage, and the genomic DNA of each plant leaf was extracted using the N96 novel plant genomic DNA extraction kit from Qiaqen corporation;
ii) breaking the genome DNA, repairing with DNA polymerase, connecting with a linker, performing electrophoresis, recovering the fragment of 300-400bp, and establishing a PE400 sequencing library;
iii) carrying out whole genome re-sequencing by using an Illumina Hiseq X10 sequencing platform and a 2X 150bp double-end sequencing method, wherein the sequencing depth of a parent is about 50X, the sequencing depth of a progeny single plant is about 0.1X, and all sequencing work can be completed within about one week;
iv) obtaining the genotype map of the single filial generation plants by carrying out whole genome sequence alignment and genetic variation analysis and identification on the single filial generation plants (each plant BC1F1 generation) and parents (Wushan silk seedlings and W3105).
And (3) screening suitable BC1F1 generation plants according to a genotype map, wherein the screening conditions are as follows: the plant contains wild rice gene segments on each chromosome; besides the wild rice gene, the other genetic background is derived from the cultivated rice genome;
in addition, in the screening, although each plant contains a wild rice gene, plants with low heterozygosity and high homozygosity are screened as much as possible;
specifically, in this example, 30 individuals of BC1F1 generation meeting the above requirements were selected.
Step 4): planting the plants of the BC1F1 generation screened in the step 3), and performing selfing to obtain seeds of the BC1F2 generation;
step 5): planting the BC1F2 generation seeds obtained in the step 4) to obtain respective plant populations, and taking leaves of the plant populations at the seedling stage to perform whole genome sequencing with low coverage rate to obtain the genotype map of each plant;
screening suitable BC1F2 generation plants under the following conditions: wild rice gene segments contained in the plant account for 5 to 20 percent of the genome background; besides the wild rice gene, other genetic backgrounds are derived from the cultivated rice genome, and the degree of purity is more than 60%; and the agronomic characters of the seedling are closer to the 'Wushan silk seedling' of the cultivated rice;
selection criteria for agronomic traits include: the plant height is 50-140 cm, the tillering number is more than 5, and the setting percentage is more than 60%; the variety which is closer to the 'Wushan silk seedling' of the cultivated rice is selected as much as possible.
Specifically, in this embodiment, 52 excellent individuals with good leaf morphology are selected, and then an individual 23 closer to the female parent is further selected from the selected individuals, which is a BC1F2 generation plant meeting the above requirements;
planting the screened proper plants of the BC1F2 generation, and performing selfing to obtain seeds of the respective BC1F3 generation;
regarding the way of selfing, bagging selfing is adopted in this embodiment.
Step 6): repeating at least 3 generations according to the mode in the step 5) until a stable new rice variety with wild rice genetic background is obtained.
Specifically, in this example, the lowest value of the fitness screening criteria in the screening conditions was increased by 10% after each selfed generation.
Specifically, seeds of BC1F3 generation are planted to obtain a plant population, and proper plants of BC1F3 generation are screened out according to the screening mode, wherein wild rice gene segments contained in the plants account for 5% -20% of genome background; besides the wild rice gene, other genetic backgrounds are derived from the cultivated rice genome, and the degree of purity is more than 70%; and 5 plants of BC1F3 generation with agronomic characters closer to the 'Wushan mountain silk seedling' of the cultivated rice;
planting 5 selected seeds of BC1F3 to obtain a BC1F4 plant group, screening proper plants of BC1F4 generation according to the screening mode and the purity of the genome background of the cultivated rice is more than 80%, simultaneously carrying out combining ability determination on the screened single plants and sterile lines II-32A, Hu family 1A, Huatai A and 9A, screening the single plants with higher combining ability, continuously bagging and selfing, and harvesting seeds (BC1F5 generation seeds);
and planting seeds of BC1F5 generation to obtain a plant group, and selecting the plants of BC1F5 generation as a stable excellent restorer line with wild rice background according to the selection mode and the purity of the genome background of the cultivated rice is more than 90 percent, wherein the restorer line is named as Huhui No. 1.
It should be understood that although the present description refers to embodiments, not every embodiment contains only a single technical solution, and such description is for clarity only, and those skilled in the art should make the description as a whole, and the technical solutions in the embodiments can also be combined appropriately to form other embodiments understood by those skilled in the art.
The above-listed detailed description is only a specific description of a possible embodiment of the present invention, and they are not intended to limit the scope of the present invention, and equivalent embodiments or modifications made without departing from the technical spirit of the present invention should be included in the scope of the present invention.
Claims (9)
1. A method for breeding a new rice variety with wild rice genetic background is characterized in that:
the method comprises the following steps of:
step 1): carrying out conventional sexual hybridization by taking a cultivated rice variety as a female parent and an AA type wild rice variety as a male parent to obtain a hybrid F1 generation;
step 2): planting the hybrid seeds obtained in the step 1) for generation F1; carrying out first backcross by taking the hybrid F1 generation as a female parent and the cultivated rice variety as a male parent to obtain BC1F1 generation seeds;
step 3): planting the BC1F1 generation seeds obtained in the step 2) to obtain a BC1F1 generation plant population, and taking leaves of the BC1F1 generation plant population at a seedling stage to perform whole genome sequencing with low coverage rate to obtain a genotype map of each BC1F1 generation plant;
screening suitable BC1F1 generation plants under the following conditions: the screened plants each contain wild rice genome segments in their genome, and the collection of different wild rice genome segments contained in these plants can cover 12 chromosomes of the wild rice genome; and, these plants, in addition to containing wild rice genomic fragments, all derive their genetic background from the oryza sativa genome;
step 4): carrying out second backcross by taking the proper plants of the BC1F1 generation screened out in the step 3) as female parents and the cultivated rice variety as male parents to obtain seeds of the BC2F1 generation;
or planting the plants of the BC1F1 generation selected in the step 3), and performing selfing to obtain seeds of the BC1F2 generation;
step 5): planting the BC2F1 generation seeds or BC1F2 generation seeds obtained in the step 4) to obtain respective plant populations, and taking leaves of the plant populations at a seedling stage to perform whole genome sequencing with low coverage rate to obtain a genotype map of each plant;
screening suitable plants of BC2F1 generation or BC1F2 generation under the following screening conditions: wild rice gene segments contained in the plant account for 5 to 20 percent of the genome background; and, except the wild rice gene contained, the genetic background is derived from the cultivated rice genome, and the screening criterion of the degree of purity is more than 60%; and, its agronomic traits more closely approximate those of the cultivar of rice;
planting the screened proper plants of the BC2F1 generation or BC1F2 generation, and performing selfing to obtain seeds of the respective next generation;
step 6): repeating at least 3 generations according to the mode in the step 5) until a stable new rice variety with wild rice genetic background is obtained.
2. The method of claim 1, wherein:
in the step 4), obtaining BC2F1 generation seeds; screening to obtain proper plants of the BC2F1 generation in the subsequent step 5), and performing selfing to obtain seeds of the BC2F2 generation; in the subsequent step 6), selfing was repeated for 3 generations to obtain stable seeds of BC2F5 generation with a wild rice genetic background.
3. The method of claim 2, wherein:
the AA type wild rice variety is '374'; the cultivated rice variety is 'WYG'.
4. The method of claim 1, wherein:
in the step 4), obtaining BC1F2 generation seeds; screening to obtain proper plants of the BC1F2 generation in the subsequent step 5), and performing selfing to obtain seeds of the BC1F3 generation; in the subsequent step 6), selfing was repeated for 2 generations to obtain stable seeds of BC1F5 generation with a wild rice genetic background.
5. The method of claim 4, wherein:
the AA type wild rice variety is W3105; the cultivated rice is Wushan silk seedling.
6. The method of any of claims 1 to 5, wherein:
in the processes of the step 5) and the step 6), after each selfing generation, the lowest value of the screening standard of the degree of purity in the screening condition is improved by 10%.
7. The method of any of claims 1 to 5, wherein:
in the screening condition in step 5), the selection criteria of the agronomic shape comprise: the plant height is 50-140 cm, the tillering number is more than 5, and the setting percentage is more than 60%.
8. The method of any of claims 1 to 5, wherein:
the whole genome sequencing with low coverage rate utilizes an Illumina Hiseq X10 sequencing platform to perform whole genome re-sequencing by a 2X 150bp double-end sequencing method, the sequencing depth of parents is about 50X, and the sequencing depth of offspring single plants is about 0.1X.
9. The method of any of claims 1 to 5, wherein:
in the step 5), the process of screening suitable plants further comprises the step of measuring the combining ability of the plants meeting the screening conditions and selecting the plants with high combining ability.
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