CN115281072B - Method for breeding high-lysine sweet and glutinous fresh corn variety - Google Patents
Method for breeding high-lysine sweet and glutinous fresh corn variety Download PDFInfo
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- A—HUMAN NECESSITIES
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Abstract
The invention relates to the technical field of waxy corn breeding, in particular to a method for breeding a high-lysine sweet and waxy fresh corn variety. The method provided by the invention creates a high-lysine sweet and waxy homozygous corn inbred line and a high-lysine waxy corn inbred line, breeds a sweet and waxy fresh-eating corn variety which simultaneously contains sweet and waxy grains and has the high-lysine character, and improves the taste quality and the nutritional quality. The ears of the sweet and waxy fresh-eating corn variety contain sweet and waxy grains, and the taste is waxy and sweet; meanwhile, the content of lysine and tryptophan is obviously improved. Through detection, the lysine content of the bred F2 ear grains is about 0.5 percent, and the tryptophan content is 0.1 percent; the lysine content is 0.4 percent higher than the national high-lysine corn standard, and the lysine content and the tryptophan content are respectively improved by more than 67 percent and 50 percent compared with the common waxy corn.
Description
Technical Field
The invention relates to the technical field of waxy corn breeding, in particular to a method for breeding a high-lysine sweet and waxy fresh corn variety.
Background
Compared with common corn, waxy corn grains contain almost all amylopectin and are sticky when eaten, so the waxy corn is also called as waxy corn. In recent years, along with the improvement of consumption level and the optimization of dietary structure, the demand of consumers on high-end high-quality and nutritious agricultural products is increasingly strengthened, the waxy corns are sticky and fragrant in taste and have no sweet taste, and the waxy corn varieties in the current market have different qualities and different taste qualities; waxy corn has rich nutrients, but low lysine and tryptophan contents. Therefore, the introduction of the sweet character and the high lysine character into the waxy corn and the improvement of the taste quality and the nutritional quality of the waxy corn can greatly meet the increasing diversified demands of people.
The waxy character, sweet character and high lysine character are all controlled by different recessive genes, and corresponding phenotypes can be presented only when the recessive genes are pure. How to efficiently aggregate the characters, overcome the interaction among genes and rapidly breed the inbred line and the variety with the simultaneously expressed characters is a big problem.
Disclosure of Invention
The invention aims to provide a method for breeding waxy corn varieties with excellent taste quality and nutritional quality at the same time, so as to improve the taste of waxy corn ears and the nutritional quality of waxy corn kernels.
The invention provides a method for breeding a high-lysine sweet and waxy fresh-eating corn variety, which comprises the following steps:
(1) Hybridizing a waxy corn backbone inbred line A and a waxy corn backbone inbred line B serving as female parents and a corn germplasm material containing opaque-2 serving as a high-lysine gene donor to obtain AF1 and BF1;
taking BF1 as a female parent and a waxy corn backbone inbred line B as a recurrent parent, and carrying out inbreeding after backcross to obtain a high-lysine waxy corn inbred line Bwxo2;
AF1 selfing to obtain a separation group, and obtaining a high-lysine waxy corn ear Awxo2 by using a molecular marker assisted method or phenotypic identification;
(2) Taking a waxy corn backbone selfing line A as a female parent and a sweet corn backbone selfing line as a sweet gene donor, selfing after hybridization to obtain ASF1, selfing and purifying the ASF1 to be used as a male parent, taking a waxy corn backbone selfing line B as a female parent to obtain hybrid progeny, measuring the ASF1 when all grains on the current hybrid ear are waxy, screening to obtain an ASF1 selfing purified single plant with strong combining ability as a male parent, hybridizing with a high-lysine waxy corn ear Awxo2, and performing phenotypic identification on the hybrid progeny to obtain a homozygous selfing line Ash2wxo2;
(3) And (3) combining the high-lysine-content sweet and waxy corn by using Bwxo2 as a female parent and using the Ash2wxo2 obtained in the step (2) as a male parent.
In the method provided by the invention, the selfing is one or more selfing generations; the backcross is one generation or more than one generation of backcross.
In the method provided by the invention, the method for obtaining the homozygous inbred line Ash2wxo2 comprises the following steps:
1) Selfing hybrid seeds by taking the waxy corn backbone selfing line A as a female parent and taking the sweet corn backbone selfing line S as a sweet gene donor, selecting sweet grains from character segregation progeny for planting, and performing phenotypic identification to obtain ASF1;
in the progeny of grain character segregation, if waxy grains are selected for planting, sweet grains can be obtained only by selfing one generation, and single plants with high germination rate and good plant growth vigor cannot be identified. According to the method, after the seed character separation progeny, the sweet seeds are selected for subsequent breeding and screening, so that the breeding period can be obviously shortened, and the single plants with high germination rate and good plant growth vigor can be rapidly obtained.
2) Selfing and purifying the ASF1 to obtain a selfed single plant, simultaneously taking a waxy corn backbone selfing line B as a female parent and taking the ASF1 as a male parent to obtain a hybrid plant, and when grains on the current-generation hybrid cluster are all waxy, keeping the corresponding selfed single plant cluster and hybrid seeds;
3) Planting the hybrid seeds retained in the step 2), carrying out accurate identification according to the requirement of variety approval, and retaining ASF1 selfing individual plant cluster numbers corresponding to the hybrid seeds with good taste quality, high yield and strong resistance;
planting the selfing single plant ears reserved in the step 2) 30-40 days after planting the hybrid seeds, performing phenotype identification, selecting ASF1 selfing single plants with high germination rate, fast seedling emergence and vigorous growth vigor, and reserving corresponding numbers;
in the invention, the hybrid planting is earlier, the inbred line selfing planting is later, and the following effects can be realized: firstly obtaining the identification result of hybrid (before the selfing line is subjected to powder scattering and spinning), obtaining a corresponding number, then selecting the selfing line with the overlapped number according to the identification result of the selfing line (obtaining the corresponding number), carrying out next selfing purification, taking the selfing line as a male parent, taking the waxy corn backbone selfing line as a female parent, and hybridizing to obtain the hybrid. If the hybrid seeds are not early, the hybrid seeds can be obtained only by carrying out hybridization in the next year, and the time is prolonged; moreover, the range of candidate inbred lines is expanded, and manpower and material resources are wasted.
4) And (3) selfing and purifying ASF1 selfing single plants existing in the two reserved numbers as male parents, and hybridizing the male parents with the high-lysine waxy corn ears Awxo2 to obtain a homozygous selfing line Ash2wxo2.
More specifically, in the method provided by the present invention, the step 4) of the method for obtaining the homozygous inbred line Ash2wxo2 is: selfing and purifying ASF1 selfing single plants existing in the two reserved numbers as male parents, hybridizing the male parents with the high-lysine waxy corn ears Awxo2 to obtain intermediate materials, selfing the intermediate materials, selecting sweet grains for planting, performing phenotype identification from the seedling stage to before pollen scattering, and identifying the single plants containing the high-lysine genes by using a molecular marker auxiliary technology; selfing the single plant, and performing phenotype identification from the seedling stage to before pollen scattering to obtain a homozygous selfing line Ash2wxo2.
In the method provided by the invention, the hybrid is accurately identified according to the requirement of variety approval, and the indexes of accurate identification comprise: the taste quality is good, the yield is high, and the resistance is strong; the phenotype identification is carried out from the seedling stage of the plant to before pollen scattering; the phenotypic identification criteria are that single plants with high germination rate, fast and regular emergence and vigorous growth are selected.
In the method provided by the invention, the growth stage from the seedling stage to the loose powder of the corn comprises the following steps: corn seedling emergence, corn jointing and bell mouth stage.
As the endosperm of the sh2 genotype grains is shriveled, the starch content is low, and the germination rate of the seeds is low; after selecting the sweet seeds, the germination rate selection of the seeds is carried out from the S1 generation, namely, strict phenotype identification is carried out from each stage from the seedling stage to the pollen scattering stage, such as seedling emergence, jointing, bellmouth and the like, and the individual plants with high germination rate, fast seedling emergence and vigorous growth vigor are selected to improve the germination rate of the seeds of the offspring.
In the method provided by the invention, in the step 1), the elimination rate of the phenotype identification is not higher than 25%; in the step 3), the elimination rate of the phenotypic identification is not less than 80%.
When the homozygous inbred line Ash2wxo2 is obtained, the sh2, wx and o2 genes are endosperm mutants, particularly, the endosperm of the sh2 genotype grains is shrunken, the starch content is low, and the seed germination rate is low; in the invention, the germination rate selection of seeds is carried out from the S1 generation, namely, strict phenotype identification is carried out at each stage from the seedling stage to before pollen scattering, such as seedling emergence, jointing, bellmouth and the like, and the individual plants with high germination rate, fast seedling emergence and vigorous growth are selected to improve the germination rate of the seeds of the offspring. In addition, the invention identifies the single plant to correspondingly select the single plant with high combining ability, breeds the excellent variety while breeding the inbred line, accelerates the variety breeding process, reduces the workload of variety matching and identification links, saves cost and time and has high efficiency.
In the method provided by the invention, the waxy corn backbone inbred line A or the waxy corn backbone inbred line B is BN2 or Jingnuo 6, N39, white nuo 6, N601, CQ56, jingnuo 2, YN-3, jingnuo 32, N203 and ZN3; the sweet gene donor is a sweet corn backbone inbred line SH251 or T68, T520, T32, T3587, T193, T18 and T29.
In the invention, the waxy corn inbred lines A and B are backbone lines, and the backbone lines have high combining ability. The new inbred line which is bred and aggregates various characters has better performance on agronomic characters, and meanwhile, the combining ability of materials can be identified in early stage through testing and matching, and the breeding efficiency is improved.
In the breeding process, the parent and the female parent used in the breeding process must simultaneously contain the high-lysine character, so that the variety with high lysine content can be bred.
As will be appreciated by those skilled in the art, the present invention provides for the use of the above-described method for improving the quality of waxy corn. And the application of the method in improving the germination rate of the sweet and waxy corn variety.
The invention has the beneficial effects that:
(1) The waxy corn variety bred by the method has improved taste quality and nutrition quality, the lysine content is more than 0.4%, the content is high, the high lysine corn standard is achieved, and the market and production requirements can be met.
(2) The invention adopts the high-lysine sweet and glutinous homozygous corn inbred line to create the sweet and glutinous corn with high lysine content, and the created high-lysine sweet and glutinous homozygous corn inbred line has good seed quality and high germination rate, and the invention breeds the high-lysine sweet and glutinous homozygous corn inbred line and simultaneously breeds an excellent variety.
Therefore, the sweet and waxy corn with high lysine content obtained by the breeding method has the advantages of short period, high success rate, high seed germination rate of the bred variety and obvious improvement of land utilization rate.
Detailed Description
The following examples are intended to illustrate the invention, but are not intended to limit the scope of the invention.
Example 1 method for breeding high-lysine sweet and waxy fresh corn variety 1
The embodiment provides a method for breeding a high-lysine sweet and waxy fresh corn variety, and the embodiment obtains the high-lysine sweet and waxy corn.
1. Obtaining high lysine waxy corn material
(1) Taking waxy corn backbone inbred lines A (BN 2) and B (Jingnuo 6) as female parents (bred and provided by corn research center of agriculture and forestry academy of sciences of Beijing), taking corn germplasm material containing opaque-2 as a high-lysine gene donor, and hybridizing to obtain AF1 and BF1; meanwhile, the A is taken as a female parent, and a self-bred sweet corn backbone inbred line SH251 of the agroforestry academy of sciences of Beijing is taken as a sweet gene donor, and the ASF1 is obtained by hybridization.
(2) And (3) taking BF1 as a female parent, taking the waxy corn backbone inbred line B as a recurrent parent, backcrossing for 4 times, and inbreeding for 2 times to obtain the high-lysine waxy corn inbred line Bwxo2, wherein the content of lysine in grains is 0.65%.
(3) AF1 is self-bred to obtain a separation population, and a molecular marker assisted method or phenotypic identification is utilized to obtain the high-lysine waxy corn ear Awxo2.
2. Obtaining high lysine sweet waxy corn material
(1) ASF1 selfing to obtain 3 kinds of phenotype grains. Selecting sweet seeds (marked as T1-T2000, S1) and planting the sweet seeds in the seedling pot by single seeds. Strict phenotype identification is carried out at each stage from the seedling stage to before pollen scattering, such as seedling emergence, jointing, bellmouth and the like, single plants with high germination rate, fast seedling emergence and vigorous growth are selected according to 20 percent elimination rate, and selfing purification is carried out; and meanwhile, taking the waxy selfing line B as a female parent, determining the selected single sweet corn plant, and when all grains on the current hybrid ear are waxy grains, keeping the corresponding selfing single plant ear, namely the cluster contains the sweet and waxy homozygous recessive gene and is recorded as TN1-TN200. While retaining the hybrid species C1-C200 obtained.
(2) Planting C1-C200, performing accurate identification according to variety approval requirements, selecting a hybridization combination with good taste quality, high yield and strong resistance, and reserving panicle rows with corresponding numbers; after 30-40 days, planting TN1-TN200 (planting according to ear rows), strictly performing phenotype identification from the seedling stage to each stage before pollen scattering, such as seedling emergence, jointing, bellmouth and the like, and selecting single plants with high germination rate, fast and regular seedling emergence and vigorous growth with a removal rate of more than 80%; combining the two aspects, identifying and selecting 20 rows of ear rows with high germination rate and high combining ability of seeds, selecting excellent individual plants TN1-TN50 from the ear rows, and performing selfing purification. Meanwhile, pollen is taken to be hybridized with Awxo2 to obtain an intermediate material.
(3) The intermediate material obtained above was planted and selfed. Selecting sweet grains for planting, performing strict phenotype identification from seedling stage to each stage before pollen scattering, such as seedling emergence, jointing, bellmouth and the like, selecting panicles with high germination rate, fast seedling emergence and vigorous growth, and identifying the single plant containing the high-lysine gene by using a molecular marker auxiliary technology. Selfing for 4 times, performing strict phenotype identification from the seedling stage to each stage before pollen scattering, such as seedling emergence, stem drawing, bell mouth and the like, selecting the ear rows with high germination rate, fast seedling emergence and vigorous growth vigor to obtain a homozygous selfing line Ash2wxo2, wherein the grain lysine content is 0.72 percent, and the seed germination rate can reach 85 percent.
3. Breeding high-lysine sweet waxy corn variety
Bwxo2 is used as a female parent, ash2wxo2 is used as a male parent, hybrid seeds are prepared, the F2 cluster contains two kinds of grains which are sweet and waxy, the mouthfeel is waxy and sweet, and the whole sugar content of the cluster is 15.2% through detection; meanwhile, the content of lysine and tryptophan is obviously improved. Through detection, the lysine content of F2 ear grains is about 0.5 percent, which is greater than the national high-lysine corn standard by 0.4 percent, and the tryptophan content is 0.1 percent, which are respectively improved by more than 67 percent and more than 50 percent compared with the common waxy corn.
Example 2 method for breeding high-lysine sweet and waxy fresh-eating corn variety 2
The difference between the method for breeding the high-lysine sweet and waxy fresh-eating corn variety and the embodiment 1 is that the waxy corn backbone inbred line A used in the embodiment is white waxy 6, and the waxy corn backbone inbred line B used in the embodiment is N39.
The Bwxo2 created in the embodiment is used as a female parent, the Ash2wxo2 is used as a male parent, hybrid seeds are prepared, the F2 cluster contains sweet and glutinous grains, the mouthfeel is glutinous and sweet, and the whole sugar content of the cluster is 15.4% through detection; meanwhile, the content of lysine and tryptophan is obviously improved. Through detection, the lysine content of F2 ear grains is 0.52 percent, is 0.4 percent higher than the national high-lysine corn standard, has the tryptophan content of 0.11 percent, and is respectively improved by more than 70 percent and 50 percent compared with the common waxy corn.
Comparative example 1
In the comparative example, a maize inbred line B (Jingnuo 6) which is the same as that in the example 1 is used as a female parent, ash2wxo2 obtained in the example 1 is used as a male parent to prepare a hybrid, an F2 cluster contains two grains of sweet and glutinous, the mouth feel is glutinous and sweet, and the whole sugar content of the cluster is 15%; through detection, the lysine content of F2 ear grains is about 0.3%, and the tryptophan content is 0.06%, which is equivalent to that of common waxy corn. Therefore, the method of comparative example 1 did not improve the nutritional quality of the resulting ears.
The results of this comparative example demonstrate that the lysine or tryptophan content of the corn ear cannot be increased by merely crossing the conventional waxy corn variety with the inbred line Ash2wxo2, i.e., the effect of improving both the taste quality and the nutritional quality of sweet waxy corn must be obtained by the method provided in example 1.
Comparative example 2
In the comparative example, bwxo2 obtained in example 1 is used as a female parent, a corn selfing line A (BN 2) same as that in example 1 is used as a male parent to assemble hybrid seeds, only waxy grains are arranged on an F2 cluster, sweet grains are not formed, the taste is not sweet, the whole sugar content of the cluster is 9%, and the cluster is equivalent to that of common waxy corn; through detection, the lysine content of F2 ear grains is 0.31%, and the tryptophan content is 0.05%, which is equivalent to that of common waxy corn.
Therefore, if the high-lysine waxy corn inbred line is adopted to be hybridized with the common waxy corn, sweet grains cannot be obtained, the lysine content or tryptophan content of the filial generation is only equivalent to that of the common corn, and the effect of improving the taste quality and the nutritional quality of the cluster cannot be achieved.
Comparative example 3
In the comparative example, the Bwxo2 obtained in the example 1 is used as a female parent, the Awxo2 obtained in the example 1 (the homozygous inbred line Awxo2 is obtained by continuous selfing and purification) is used as a male parent, and the hybrid is prepared, and through detection, the lysine content and the tryptophan content of F2 ear grains are 0.45% and 0.09% which are higher than those of common waxy corns, but the F2 ears are all waxy corn grains, so that the mouth feel is sweet, the whole sugar content of the ears is 8.6%, and the corn grain is equivalent to that of the common waxy corns.
Thus, comparative example 3 demonstrates that the cluster taste quality of the resulting sweet waxy maize is not improved if the Awxo2 is not further optimized.
Comparative example 4
1. Obtaining high lysine waxy corn material
(1) Taking waxy corn backbone inbred lines A (BN 2) and B (Jingnuo 6) as female parents (bred and provided by corn research center of agriculture and forestry academy of sciences of Beijing), taking corn germplasm material containing opaque-2 as a high-lysine gene donor, and hybridizing to obtain AF1 and BF1; meanwhile, the A is taken as a female parent, and a self-bred sweet corn backbone inbred line SH251 of the agroforestry academy of sciences of Beijing is taken as a sweet gene donor, and the ASF1 is obtained by hybridization.
(2) And (3) backcrossing 4 times and selfing 2 times by taking BF1 as a female parent and the waxy corn backbone selfing line B as a recurrent parent to obtain a high-lysine waxy corn selfing line Bwxo2, wherein the content of lysine in grains is 0.65%.
(3) AF1 is self-bred to obtain a separation population, and a molecular marker assisted method or phenotypic identification is utilized to obtain the high-lysine waxy corn ear Awxo2.
2. Obtaining high lysine sweet waxy corn material
(1) ASF1 selfing to obtain 3 kinds of phenotype grains. Selecting sweet seeds (marked as T1-T2000, S1) and planting the sweet seeds in the seedling pot by single seeds. Taking single plant leaves, identifying by using a molecular marker technology to obtain homozygous recessive single plants containing sh2 and wx, and selfing and purifying. Meanwhile, pollen is taken to be hybridized with the Awxo2 material to obtain an intermediate material.
(2) Selfing the intermediate material, taking leaves at the seedling stage, identifying by using a molecular marking method to obtain a recessive homozygous material containing sweet, glutinous and high lysine, and selfing to obtain a homozygous selfing line Ash2wxo2. The germination rate of the seeds is 50 percent.
Therefore, the method of comparative example 4 can obtain a homozygous inbred line with a significantly reduced seed germination rate compared to example 1.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, it is intended that all such modifications and alterations be included within the scope of this invention as defined in the appended claims.
Claims (8)
1. A method for breeding a high-lysine sweet and glutinous fresh corn variety is characterized by comprising the following steps:
(1) Hybridizing a waxy corn backbone inbred line A and a waxy corn backbone inbred line B serving as female parents and a corn germplasm material containing opaque-2 serving as a high-lysine gene donor to obtain AF1 and BF1;
taking BF1 as a female parent and a waxy corn backbone inbred line B as a recurrent parent, and carrying out inbreeding after backcross to obtain a high-lysine waxy corn inbred line Bwxo2;
AF1 is selfed to obtain a separation population, and a high-lysine waxy corn ear Awxo2 is obtained by a molecular marker assisted method or phenotypic identification;
(2) The method for obtaining the homozygous inbred line Ash2wxo2 comprises the following steps:
1) Selfing hybrid seeds by taking the waxy corn backbone selfing line A as a female parent and taking the sweet corn backbone selfing line as a sweet gene donor, selecting sweet grains from character segregation progeny for planting, and performing phenotypic identification to obtain ASF1;
2) Selfing and purifying the ASF1 to obtain a selfed single plant, simultaneously taking a waxy corn backbone selfing line B as a female parent and taking the ASF1 as a male parent to obtain a hybrid plant, and when grains on the current-generation hybrid cluster are all waxy, keeping the corresponding selfed single plant cluster and hybrid seeds;
3) Planting the hybrid seeds retained in the step 2), carrying out accurate identification according to variety approval requirements, and retaining ASF1 selfing individual plant ear numbers corresponding to the hybrid seeds with good taste quality, high yield and strong resistance;
planting the selfing single plant clusters reserved in the step 2) 30-40 days after the hybrid seeds are planted, performing phenotype identification, selecting ASF1 selfing single plants with high germination rate, fast seedling emergence and vigorous growth, and reserving corresponding numbers;
4) Selfing and purifying ASF1 selfing single plants existing in the two reserved numbers as male parents, and hybridizing the male parents with the high-lysine waxy corn ears Awxo2 to obtain a homozygous selfing line Ash2wxo2;
(3) Preparing a high-lysine sweet and waxy fresh corn variety by using Bwxo2 as a female parent and using Ash2wxo2 obtained in the step (2) as a male parent;
the waxy corn backbone inbred line A or the waxy corn backbone inbred line B is BN2 or Jingnuo 6, N39, white nuo 6, N601, CQ56, jingnuo 2, YN-3, jingnuo 32, N203 and ZN3; the sweet gene donor is a sweet corn backbone inbred line SH251 or T68, T520, T32, T3587, T193, T18 and T29.
2. The method of claim 1, wherein the selfing is a selfing generation or more; the backcross is one generation or more than one generation of backcross.
3. The method according to claim 2, wherein step 4) is: selfing and purifying ASF1 selfing single plants existing in the two reserved numbers as male parents, hybridizing the male parents with high-lysine waxy corn ears Awxo2 to obtain intermediate materials, selfing the intermediate materials, selecting sweet grains for planting, performing phenotype identification from the seedling stage to before pollen scattering, and identifying the single plants containing high-lysine genes by using a molecular marker auxiliary technology; selfing the single plant, and performing phenotype identification from the seedling stage to before pollen scattering to obtain a homozygous selfing line Ash2wxo2.
4. The method of claim 3, wherein the hybrid is precisely identified according to the requirements of variety approval, and the precise identification indexes comprise: the taste quality is good, the yield is high, and the resistance is strong; the phenotypic identification criteria are that single plants with high germination rate, fast and regular emergence and vigorous growth are selected.
5. The method of claim 4, wherein the corn growing stage from the seedling stage to the pre-fluffing stage comprises: corn seedling emergence, corn jointing and bell mouth stage.
6. The method of claim 5, wherein in step 1), the elimination rate of phenotypic identification is not higher than 25%; in the step 3), the elimination rate of the phenotype identification is not lower than 80%.
7. Use of the method of any one of claims 1-6 to increase lysine and tryptophan content in waxy maize.
8. Use of the method of any one of claims 1-6 to increase germination rate of a sweet waxy corn variety.
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