CN114747482A - Breeding method of high-combining-ability sweet-waxy double-cryptocorn inbred line - Google Patents
Breeding method of high-combining-ability sweet-waxy double-cryptocorn inbred line Download PDFInfo
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Abstract
The invention discloses a breeding method of a high-combining ability sweet-waxy double-hidden corn inbred line, which comprises the steps of hybridizing corn materials with sweet and waxy genes, constructing an original population, selecting an excellent single plant for inbreeding to obtain F2Seeds are substituted, sweet grains are selected for planting, individual plants with waxy genes are screened by utilizing molecular markers, and individual plants with waxy genes and excellent comprehensive characters are selected for selfing to obtain S1Seed generation and planting S1Seed generation to obtain S1And (4) generating a strain, selecting an excellent plant from the strain for selfing, hybridizing the rest pollen with the waxy corn selfing line, further screening the strain according to the hybridization combination expression, repeating the steps, and performing shuttle breeding in Sichuan and Yunnan to realize the synchronization of the combination breeding of the excellent selfing line and the sweet waxy corn. The sweet and glutinous double-hidden corn inbred line bred by the method has the advantages of good quality, wide adaptability, high combining ability and the like, and simultaneously hybrid seeds can be combined in early generations, so that the excellent quality can be effectively improvedThe combination efficiency of the good combination is improved, and the breeding time is shortened.
Description
Technical Field
The invention belongs to the technical field of breeding of new agricultural varieties, and particularly relates to a breeding method of a sweet and waxy double-cryptic corn inbred line with high combining ability.
Background
The sweet and waxy corn is popular with consumers due to the unique flavor, and particularly the sweet and waxy corn meets the requirements of people on the sweet and waxy fresh corn. The sweet and waxy corn is a cultivation type artificially created and contains sweet and waxy grains on the same corn ear, the corn variety is obtained by cultivating a sweet and waxy double-implicit corn self-bred line and a waxy corn self-bred line to hybridize to obtain F1, and the sweet grains and the waxy grains exist on the fruit ear of an F1 plant. In the breeding of the sweet and waxy corns, the sweet and waxy double-cryptocorn inbred line is the basis of the breeding of the sweet and waxy corns.
The problems of narrow germplasm resources, weak genetic basis, long breeding period, low efficiency and the like of the existing sweet and waxy double-recessive maize inbred line are obvious, so that the breeding efficiency of the inbred line is improved, the germplasm resource innovation of the sweet and waxy maize is enhanced, and the cultivation of the high-combining ability sweet and waxy double-recessive maize inbred line is of great significance.
CN104521742A discloses a method for rapidly creating a sweet and glutinous double recessive corn homozygous inbred line, which comprises the following steps: the method comprises the following steps of hybridizing sweet corn and waxy corn, collecting F1 seeds, hybridizing a haploid induction line with a purple marker by taking F1 as a female parent to obtain haploid seeds, doubling and selfing the haploid to obtain a powdery corn double-single line, a waxy corn double-single line and a sweet corn double-single line, and identifying a sweet material by utilizing a molecular marker linked with wx genes to obtain a sweet and waxy double-recessive selfing line material. Although the technology can rapidly create the sweet and waxy double recessive corn homozygous inbred line, the sweet and waxy double recessive corn inbred line with high combining ability cannot be accurately obtained.
Disclosure of Invention
Aiming at the problems, the invention provides a breeding method of a high-combining-ability sweet-waxy double-cryptocorn inbred line. The method comprises hybridizing corn material with sweet and waxy genes, constructing original population, selecting excellent individual plant, and selfing to obtain F2Seeds are substituted, sweet grains are selected for planting, individual plants with waxy genes are screened by utilizing molecular markers, and individual plants with waxy genes and excellent comprehensive characters are selected for selfing to obtain S1Seed generation and planting S1Seed generation to obtain S1And (4) generating a strain, selecting an excellent plant from the strain for selfing, hybridizing the rest pollen with the waxy corn selfing line, further screening the strain according to the hybridization combination expression, repeating the steps, and performing shuttle breeding in Sichuan and Yunnan to realize the synchronization of the combination breeding of the excellent selfing line and the sweet waxy corn. The sweet and glutinous double-hidden corn inbred line bred by the method has the advantages of good quality, wide adaptability, high combining ability and the like through alternate identification and breeding of years and different ecological regions, and meanwhile hybrid seeds are combined in early generations, so that the combination efficiency of excellent combinations can be effectively improved, and the breeding time is shortened.
The invention aims to realize the following technical scheme: a breeding method of a high-combining ability sweet-waxy double-hidden corn inbred line comprises the following steps:
s1: hybridization to obtain F1Seed generation
Respectively selecting excellent sweet corn material and glutinous corn material, and hybridizing the glutinous corn material and the sweet corn material to obtain F1Seed generation (common powdery seeds);
s2: selfing to obtain F2Seed generation
Planting F1Seed generation and selfing to obtain F2Seed generation (obtaining sweet, glutinous and powdery seeds), and selecting sweet seeds;
s3: obtaining sweet and glutinous double-hidden corn single plant S1Seed generation
Seeding sweet in step S2Selecting sweet and glutinous double-hidden corn single plants containing glutinous genes by adopting molecular markers during seedling stage sampling, selecting the single plants, bagging and selfing, selecting the sweet and glutinous double-hidden corn single plants without obvious defects in comprehensive characters during harvesting, and obtaining S1Seed generation;
s4: high combining ability sweet and glutinous double-hidden corn self-bred line and sweet and glutinous corn combined breeding are carried out synchronously
Planting S1Seed generation to obtain S1Selecting the best strain from each strain for selfing, hybridizing the residual pollen with the selfing line of waxy corn, and further screening by the hybridization combination expression1And (4) carrying out shuttling breeding in two different ecological areas (such as Sichuan and Yunnan) by repeating the steps in the way, and realizing the synchronization of the high-combining-ability sweet-waxy double-hidden corn inbred line and the sweet-waxy corn combined breeding.
Further, in step S1, sweet and glutinous corn materials with excellent comprehensive properties, good quality and long harvest time are selected.
Further, the new combination of the bred sweet and waxy corn has sweet grains on the first filial generation of cluster: waxy seeds are 1: 3.
Further, the molecular label in step S3 is: molecular markers and corresponding primer sequences closely linked to or co-segregating with the waxy genes can be screened using data published by MaizeGDB, preferably using marker Phi 061.
Furthermore, the self-bred line of the backbone waxy corn in the step S4 is selected to meet the requirements of high general mating force and large genetic difference, and belongs to different heterosis groups.
The step S4 can largely eliminate the strains with poor early expression, concentrate the strength to select the best strain from the best strains, and simultaneously, the strains are alternately planted in Sichuan and Yunnan, and the identification can be carried out in different ecological areas, thereby increasing the breeding probability of the excellent sweet and glutinous double-hidden corn inbred line.
Further, the step S4 specifically includes:
(1) planting S1Seed generation to obtain S1Generation line, selecting excellent plants from the line to self-copulate to obtain S2Seed generation, hybridizing the residual pollen with 3-5 backbone waxy corn inbred lines to obtain S2Generation hybridization combination;
(2) s in planting step (1)2Seed generation to obtain S2Generating strain lines;
(3) s in the planting step (1)2Combining generations, harvesting fresh ears, evaluating the quality and yield characters of each combination, calculating the combining ability and yield contrast advantages, and selecting a combination with high combining ability and yield contrast advantages; eliminating S with poor character combining ability performance in the step (2) according to screening results2Generation line to obtain S with high combining ability2Generating strain lines;
(4) s after screening2Selecting excellent plants from the excellent generation strains for selfing to obtain S3Seed generation;
(5) will S3Repeating the steps (1) to (3) for the seeds to obtain the S with high combining ability4Generation line and yield contrast advantage high S4And (4) generation hybridization combination.
Further, S with high fitting force can be used4The generation lines continue to self-cross and are directionally assembled to S in the step (5)4And (4) generation of hybrid combination, and timely upgrading into a new variety screening test.
The breeding method of the high-combining ability sweet-waxy double-cryptocorn inbred line is schematically shown in figure 1.
Compared with the prior art, the invention has the following advantages and effects:
(1) wide selection of base material
The invention has no limitation on the selection of the basic materials, and sweet and waxy gene donor materials can be selected from groups, commercial hybrids, inbred lines and the like according to breeding targets.
(2) Improve the breeding probability of the excellent sweet and waxy double-hidden corn inbred line
The invention can eliminate a great deal of plants with poor comprehensive properties and low combining ability in the early period, concentrate on selecting excellent plants from the excellent lines and improve the breeding efficiency of the excellent inbred lines.
(3) Improve the mating efficiency of the hybrid
The invention can be used for matching hybrid seeds in the early stage, screening combinations with good performance, and selecting specific strains for directional matching in the later generation, thereby improving the matching efficiency of excellent combinations
(4) Shortening breeding period
By utilizing the invention, the breeding of the hybrid and the breeding of the inbred line are synchronously carried out, and the breeding period can be effectively shortened.
(5) Alternate identification and breeding of different ecological regions
The sweet-waxy double-hidden corn inbred line bred by the method has the advantages of good quality, wide adaptability, high combining ability and the like through alternate identification breeding of years and different ecological regions.
Drawings
FIG. 1 is a flow chart of a breeding method of a high combining ability sweet and waxy double-cryptic corn inbred line;
FIG. 2 shows the plants (top) and ears (bottom) of the inbred lines NT123, NT213 and NT421 of sweet and waxy bicontinuous maize; from left to right in sequence: the sweet and waxy double-cryptocorn inbred lines NT123, NT213 and NT 421.
Detailed Description
The present invention will be further described with reference to the following examples and accompanying drawings. The DNA extraction method, SLS, used in the following examples, included the following steps:
(1) shearing a small amount of leaves, putting in liquid nitrogen, grinding into powder, quickly transferring into a 2ml EP tube, adding 800 μ l of SLS extract, shaking for 5 min, and shaking uniformly; (2) equal volume of phenol was added: chloroform: isoamyl alcohol (25:24:1), shaking for 5 minutes, and centrifuging at 12000rpm for 10 minutes after shaking uniformly; (3) sucking 600. mu.l of the supernatant into a new 1.5ml centrifuge tube, adding equal volume of precooled isopropanol (-20 ℃) to precipitate DNA; (4) centrifuging at 12000rpm for 10min, pouring out supernatant, washing with 75% ethanol once, rinsing with pure ethanol once, air drying, adding pure water, and placing in a refrigerator at-20 deg.C for use. SLS extracting solution formula: 100ml of 0.2M EDTA, 100ml of 1mol/L Tris-HCl, 100ml of 1M NaCl and 10g of SLS, and the volume is up to 1000 ml.
Three diaphyseal waxy maize inbreds used in the examples below: (1) white 478 WR-1: is prepared by hybridizing a common corn inbred line fluid 478 and a local waxy corn variety, and performing continuous 8-generation inbred breeding; QN21 and QN24 are obtained by continuous selfing and breeding of Zhejiang waxy corn No. 4 and Suke waxy corn No. 11.
Example 1
(1) The double hybrid JY01 (F) is prepared by taking Jingke glutinous 2000 as a female parent (P1) and taking normal sweet 68 as a male parent (P2)1)，F1The grains are represented as common powdery grains (Sichuan/spring);
(2) planting JY01, selecting 10 plants in flowering period, bagging and selfing to obtain F2Generation seeds, namely JY01-1, JY01-2, JY01-3, JY01-4, JY01-5, JY01-6, JY01-7, JY01-8, JY01-9 and JY01-0 (Yunnan/winter);
(3) separating powder, glutinous and sweet grains from the grain type, selecting F2Planting sweet grains in generation seeds, cutting leaves at seedling stage, strictly numbering, wherein the plant number is the same as that of the leaves, extracting DNA by adopting an SLS method for sampling the leaves, screening sweet and glutinous double-hidden corn single plants containing glutinous genes by using a Phi061 marker, directionally removing the single plants not containing the glutinous genes in the field according to experimental results, bagging and selfing all the other single plants, identifying the comprehensive characters in the field during harvesting, and finally selecting 40 sweet and glutinous double-hidden corn single plants without obvious defects in the comprehensive characters to obtain S1Seed generation (Sichuan/spring);
(4) planting S1Seed generation to obtain 40S1Generation lines, selecting the best plants from each line to self-cross to obtain S2Generating seeds, hybridizing the rest pollen with 3 stem waxy corn inbred lines (white 478WR-1, QN21 and QN24) to obtain 312S seeds2Generation combination, wherein selfing single plants and combination are strictly numbered (Yunnan/winter);
(5) planting S2Seed generation to obtain 92S2Generation lines (Sichuan/spring);
(6) planting 312S2Combining generation, designing according to random block experiment, repeating for three times, performing single-row area, each row has 10 pits, setting seedling in each pit with density of 3200 plants/mu, harvesting fresh ear 20-25 days after silking, evaluating quality and yield of each combination, and calculating combining ability and yield control advantage (combining ability calculating method can refer to Sprague)&Tatum), selecting lines with high combining ability and combinations with high yield contrast advantage according to calculation results, and finally screening 25 high-yield and high-quality sweet and glutinous corn combinations and high-combining ability corn S231 generation lines (Sichuan/spring);
(7) according to the strain screening result in the step (6), strains with poor character combining ability performance in the step (5) are eliminated, advantages are concentrated on the excellent strains to select the excellent strains for selfing, 53 excellent single strains are harvested, and S is obtained3Seed generation; (Sichuan/spring)
(8) Planting S3Seed generation to obtain 53S3Generation lines, selecting the best plant in each line to self-cross to obtain S4Seed generation, hybridizing the rest pollen with waxy maize inbred line (white 478WR-1, QN21, QN24) to obtain 208S4Generation combination, wherein selfing single plants and combination are strictly numbered (Yunnan/winter);
(9) respectively planting S4Seeds and combinations are generated, and strains and combinations are strictly screened according to combination performance (the method is the same as the step (6)), so that 3 sweet and waxy double-hidden corn strains with high combining ability and 7 strong-dominance corn combinations (Sichuan/spring) are obtained;
(10) 3S' S best performing in the planting step (9)4The seed of the preferred single plant of the generation line is obtained to obtain S5And generating strains, directionally combining 7 strong dominant corn combinations, and timely upgrading and participating in a new variety screening test (Yunnan/winter).
By utilizing the method, 3 sweet and waxy double-cryptic corn inbred lines NT123, NT213 and NT421 (figure 2) with high combining ability and 7 strong-superiority corn combinations are obtained by co-breeding.
Claims (8)
1. A breeding method of a high-combining ability sweet-waxy double-cryptocorn inbred line is characterized by comprising the following steps:
s1: hybridization to obtain F1Seed generation
Respectively selecting excellent sweet corn material and glutinous corn material, and hybridizing the glutinous corn material and the sweet corn material to obtain F1Seed generation;
s2: selfing to obtain F2Seed generation
Planting F1Seed generation and selfing to obtain F2Replacing seeds, and selecting sweet seeds;
s3: obtaining sweet and glutinous double-hidden corn single plant S1Seed generation
Sowing the sweet seeds in the step S2, and the seedling stageSampling, screening sweet and glutinous double-hidden corn single plants containing glutinous genes by adopting molecular markers, selecting the single plants, bagging and selfing, selecting the sweet and glutinous double-hidden corn single plants without obvious defects in comprehensive characters during harvesting, and obtaining S1Seed generation;
s4: high combining ability sweet and glutinous double-hidden corn self-bred line and sweet and glutinous corn combined breeding are carried out synchronously
Planting S1Seed generation to obtain S1Selecting the best strain from each strain for selfing, hybridizing the residual pollen with the selfing line of waxy corn, and further screening by the hybridization combination expression1And (4) carrying out shuttling breeding in two different ecological areas by repeating the generation line, so as to realize the synchronization of the sweet and waxy double-hidden corn inbred line with high combining ability and the sweet and waxy corn combined breeding.
2. The breeding method of the high combining ability sweet and waxy double-cryptic corn inbred line of claim 1, which is characterized in that the step S4 specifically comprises the following steps:
(1) planting S1Seed generation to obtain S1Generation line, selecting excellent plants from the line to self-copulate to obtain S2Seed generation, hybridizing the residual pollen with 3-5 backbone waxy corn inbred lines to obtain S2Generation hybridization combination;
(2) s in planting step (1)2Seed generation to obtain S2Generating strain lines;
(3) s in the planting step (1)2Combining generations, harvesting fresh ears, evaluating the quality and yield characters of each combination, calculating the combining ability and yield contrast advantages, and selecting a strain with high combining ability and a combination with high yield contrast advantages; eliminating S with poor character combining ability performance in the step (2) according to screening results2Generation line to obtain S with high combining ability2Generating strain lines;
(4) s after screening2Selecting excellent plants from the excellent generation strains for selfing to obtain S3Seed generation;
(5) will S3Repeating the steps (1) to (3) for the seeds to obtain the S with high combining ability4Generation line and yield contrast advantage high S4And (4) generation hybridization combination.
3. The method for breeding the high combining ability sweet and waxy double-cryptic corn inbred line as claimed in claim 2, which is characterized in that the breeding method of the high combining ability S4The generation lines continue to self-cross and are directionally assembled to S in the step (5)4And (4) generation of hybrid combination, and timely upgrading into a new variety screening test.
4. The breeding method of the high combining ability sweet and waxy double-hidden corn inbred line according to any one of claims 1 to 3, characterized in that two different ecoregions are Sichuan and Yunnan.
5. The method for breeding the high combining ability sweet and waxy double-cryptic corn inbred line of any one of claims 1-3, which is characterized in that F in step S11Sweet seeds on the combined cluster of the sweet and waxy corn: waxy seeds are 1: 3.
6. The breeding method of the high combining ability sweet and waxy double-cryptic corn inbred line of any one of claims 1-3, characterized in that the molecular markers in step S3 are: molecular markers closely linked to or co-segregating with the waxy gene and corresponding primer sequences were screened using data published by MaizeGDB.
7. The breeding method of the inbred line of the sweet-waxy double-cryptocorn with high combining ability as claimed in claim 6, wherein the molecular marker is molecular marker Phi 061.
8. The method for breeding the sweet and waxy double-cryptic corn inbred line with high combining ability as claimed in any one of claims 1 to 3, wherein the step S4 backbone waxy corn inbred line has high combining ability and large genetic difference and belongs to different heterosis groups.
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Cited By (2)
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| CN116686697A (en) * | 2023-07-07 | 2023-09-05 | 云南省农业科学院粮食作物研究所 | Method for improving fresh waxy corn into fresh sweet waxy corn |
| CN116724882A (en) * | 2023-07-07 | 2023-09-12 | 云南省农业科学院粮食作物研究所 | Method for cultivating multi-spike small waxy corn with 8-10 spike rows |
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