CN113678730A - Breeding method of fresh-eating corn with crisp and tender pericarp - Google Patents
Breeding method of fresh-eating corn with crisp and tender pericarp Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
Abstract
The invention discloses a breeding method of fresh corn with crisp and tender pericarp, which comprises the following steps: (1) sowing the breeding original material (2) to induce single grains, wherein the method for obtaining the induced single grains comprises two methods of screening firstly and then inducing and inducing firstly and then screening; sowing the seeds and inducing single seeds, and selfing to obtain induced 1-generation fruit clusters; the brittleness detection screening method comprises the following steps: the corn kernel in the fresh-eating period is taken as a detection sample, the crispness and the tenderness are measured by the force (unit: cattle) for puncturing the peel by a measuring needle with the diameter of 1.0mm, and the measured value is less than 2.1N. (3) And (5) detecting pericarp and identifying phenotype to obtain a pericarp crisp and tender type fresh-eating corn inbred line. Compared with the common fresh corn, the cooking quality of the fresh corn with crisp and tender pericarp is obviously improved, the measured value of the pericarp strength is obviously reduced, the palatability is greatly improved, the pericarp is crisp and tender, and the mouthfeel has no pericarp residues; by combining the pericarp crisp and tender type inbred line and the non-crisp and tender type inbred line, the pericarp residue degree can be obviously improved.
Description
Technical Field
The invention relates to the field of crop breeding, in particular to a breeding method of fresh-eating corn with crisp and tender pericarp.
Background
In fresh corn, palatability is the most critical index for measuring the quality of varieties. Palatability, also known as mouth feel, as the name suggests, refers to the sensation of being eaten in the mouth. Regarding the palatability of fresh corn, China defines the palatability as cooking quality and sets corresponding indexes, wherein the indexes mainly comprise: sweetness, glutinousness, tenderness, smell, flavor, color and thickness of the skin. Among these indexes, sweetness and glutinousness are undoubtedly one of important indexes, but due to simple inheritance, abundant resources and relatively easy character breeding. The peel is controlled by multiple genes and has complex genetic relationship, and the palatability of the peel is difficult to improve. Therefore, the core of the quality improvement of the fresh corn is the improvement of the pericarp in a certain sense. The palatability of the peel is the feeling of pomace when chewed. The variety with good palatability of the peel gives people a soft, crisp and tender feeling without peel residue and fiber when chewing.
The pericarp is the organ that is first established and developed, relative to the endosperm. On waxy corn, the endosperm is not fully filled in the fresh eating period, and the dry matter content is relatively less, so the feeling of skin and slag is prominent when the waxy corn is eaten. When some varieties are mature, the feeling of skin dregs is relatively reduced along with the filling of endosperm, but the skin dregs are more prominent after the skins of some varieties are aged. On sweet corn, because the endosperm contains almost no starch, less dry matter and more water, the proportion of the pericarp is relatively large, and the feeling of the pericarp and the dreg is more obvious than that of waxy corn, especially the super-sweet corn. The problem is more pronounced as the skin ages when it is mature.
At present, the quantitative indexes for measuring the degree of the fresh corn peel residue at home and abroad mainly comprise the peel residue rate and the peel thickness. The skin residue rate is the percentage of the skin residue in the whole kernel in the fresh food period; the thickness of the peel is the microscopically measured thickness value of the peel in the fresh-eating period. Many sweet corn and waxy corn studies show that the pomace rate and palatability are in positive correlation with the thickness of the skin, namely the thicker the skin, the higher the pomace rate and the heavier the pomace feeling. The rate of the pomace is influenced by the thickness of the pericarp and also relates to the size of the corn kernels, and the smaller the particles, the larger the surface area of the corn kernels under the unit weight, namely the higher the rate of the pomace. In both fresh corn breeding and production, it is desirable that the thinner the skin the better. Thus, breeders breed in the direction of the thin skin in many ways. Research shows that the thickness and the peel content of the seed coat are greatly different among different varieties, and the peel generally accounts for 6-10% of the dry weight of the whole seed coat. The pericarp thickness of maize is controlled by multiple genes, and studies suggest that at least 8 pairs of genes are involved.
However, the thickness of the corn husk is limited in biological terms. However, with the improvement of living standard of people, the quality requirement of people on fresh corn is higher and higher, and the limited germplasm resources are added, so the difficulty of pericarp breeding is higher and higher.
The research finds that the tasting identification score of the cooking quality of the fresh corn is not completely related to the rate of the husk dreg and the thickness of the husk (negative correlation). In some tasting samples, it is common to have a high skin scum rate and skin thickness measurement, but not necessarily a low tasting score. It is concluded that corn pericarp may differ in crispness, which results in higher pomace content and still better palatability scores for varieties with thicker pericarp. Inspired by the research, the research develops a new way and decides to develop the research from the direction of crisp and tender of the peel so as to solve the worldwide problem that the fresh corn, especially the sweet corn, has thick peel and much peel and residue.
Disclosure of Invention
In order to make up the defects of the prior art, the invention aims to provide a breeding method of fresh-eating corn with crisp and tender pericarp, which utilizes the pericarp strength value to screen the crispness and tenderness of the pericarp, has quantitative and direct indexes, closely related detection values and palatability and good selection effect; the inbred line and the hybrid of the fresh corn with crisp and tender pericarp are bred, the taste palatability (cooking quality) in the fresh food period is obviously improved, and the taste is crisp and tender without the skin residue; the strength of the peel in the fresh eating period and the mature period is obviously reduced.
The invention aims to realize the breeding method of the fresh corn with crisp and tender pericarp by adopting the following technical scheme, which is characterized by comprising the following steps of: the method comprises the following steps:
(1) original material for breeding
The breeding original material comprises a separation generation of a fresh corn hybrid, the fresh corn hybrid comprises various varieties of sweet corn, super-sweet corn and waxy corn, the separation generation comprises separation generations of F2, F3 to Fn, or semi-finished material in a self-bred line breeding process comprises S1, S2, S3 to SN, or is obtained by mixed sowing and mixed pollination of a plurality of fresh corn hybrid varieties;
(2) sowing induced single grain
The method for obtaining the induced single particles comprises two methods of screening firstly and then inducing and inducing firstly and then screening; sowing the seeds and inducing single seeds, and selfing to obtain induced 1-generation fruit clusters;
the method for screening firstly and inducing secondly comprises the following steps:
the raw material is an inbred line, and the breeding raw material is subjected to brittleness detection and screening to obtain crisp and tender corn seeds; performing single-seed sowing on the crisp and tender corn seeds, performing induction line selection on the sowed ear rows or single plants according to ear rows or mixed sowing, and obtaining induced single seeds through diploid induction;
the method for inducing firstly and screening secondly comprises the following steps:
the raw material is hybrid seeds and comprehensive seeds; obtaining current-generation induced seeds through diploid induction; performing brittleness detection screening to obtain induced single grains;
the brittleness detection screening method comprises the following steps: the corn kernel in the fresh-eating period is taken as a detection sample, the crispness and the tenderness are measured by the force (unit: cattle) for puncturing the peel by a measuring needle with the diameter of 1.0mm, and the measured value is less than 2.1N.
c. Pericarp detection and phenotype identification:
performing brittleness detection on the induced 1-generation fruit cluster, wherein the genotype of the induced 1-generation fruit cluster is homozygous, and the genotype is consistent with the phenotype, so that a pericarp crisp and tender type fresh-eating corn inbred line is obtained;
the brittleness detection method is the same as the brittleness detection screening method in the step (2); the fruit peel strength value of the fresh-eating period fruit peel of the crisp and tender type fresh-eating corn inbred line is less than 2.1N, the fruit peel at the top end of mature seed grains has wrinkles, and endosperm, embryos and other parts are normal.
Furthermore, the brittleness detection method adopts a corn peel hardness tester for testing, and the corn peel hardness tester is a fresh corn peel hardness tester (the patent number is ZL 201920602949.7); sampling in the fresh eating period, breaking off 1/3-1/2 of the upper part of the cluster for determination, reserving seeds on the rest cluster, covering a pollination paper bag for protection, preventing water from entering and rotting, and taking 10-15 grains with regular shapes at the middle part of each cluster.
Further, inducing 1-generation selfing to obtain induced 2-generation ear rows, identifying the phenotypic purity of the field plants, eliminating poor characters, simultaneously performing pericarp brittleness determination on the selected ear rows again, namely checking, and finally obtaining homozygous ear rows, namely obtaining the pericarp brittle and tender type maize inbred line.
Further, the pericarp crisp and tender type fresh-eating corn inbred line is used for preparing crisp and tender pericarp hybrids, and the specific method comprises the following steps: the pericarp crisp and tender type fresh-eating corn inbred line is multiplied by the pericarp crisp and tender type fresh-eating corn inbred line to prepare the crisp and tender type pericarp fresh-eating corn hybrid, or the pericarp crisp and tender type fresh-eating corn inbred line is multiplied by the non-pericarp crisp and tender type fresh-eating corn inbred line to prepare the semi-crisp and tender type fresh-eating corn hybrid.
The technology has the following good effects:
1. the inbred line and the hybrid of the fresh corn with crisp and tender pericarp are bred, the taste palatability (cooking quality) in the fresh food period is obviously improved, and the taste is crisp and tender without the skin residue; the strength of the peel in the fresh eating period and the mature period is obviously reduced.
2. The thickness of the peel and the height of the peel residue rate do not need to be considered, the peel brittleness and tenderness are screened by using the peel strength value, the indexes are quantized and direct, the detection value is closely related to the palatability, and the selection effect is good.
3. The application of the diploid induction technology solves the problem of difficult line selection caused by inconsistent phenotype and genotype of the fruit peel and the separation of the endosperm and the embryo in one generation, the phenotype is equal to the genotype, and the selection result is accurate.
4. Compared with the micro-measurement of the peel thickness and the peel residue rate, the peel strength value detection is simpler and easier to operate, can detect seeds in the fresh food period and the mature period, and brings convenience to breeding operation.
5. The peel character is genetically separated by taking the ear rows as units, so a large number of ear rows need to be sown in the line selection process. The application of the diploid induction technology not only greatly reduces the number of self-crossing generations, but also greatly reduces the ear-row planting, and greatly reduces the land area and the cost.
6. The breeding and discovery of the germplasm of the crisp and tender fresh-eating corn open up a new way for solving the worldwide problem of the skin residue of the fresh-eating corn, in particular the super-sweet corn. Meanwhile, the breeding of the new germplasm provides a new gene for exploring the physiological mechanism and gene mining and even transformation of the quality of fresh corn.
Drawings
FIG. 1 is a technical scheme of the invention.
Detailed Description
1. Original material for breeding
Various fresh-eating corn test materials, including fresh-eating corn such as sweet corn, waxy corn, super-sweet corn and the like, hybrid seeds, groups, comprehensive seeds and inbred lines, F1, F2 and the like, can be used as breeding test materials. In order to enrich the genetic diversity of breeding test materials, a plurality of varieties (original materials) can be sown in a mixed mode and pollinated in a mixed mode.
2. Measurement index
The intensity value (unit N, cattle) of the corn peel is measured, the crispness of the corn peel is measured by adopting a fresh corn peel hardness tester (ZL 201920602949.7) invented before, the force (unit: cattle) used for puncturing the corn peel by a measuring needle with the diameter of 1.0mm is used for expressing the crispness of the corn peel, the measured value is less than 2.1N and is taken as the lowest standard of the crispness, the lower the value is, the better the crispness is, and the lower the crispness is, otherwise, the crispness is poor.
3. Generation of assay and sampling
(1) The generation was measured. The test material may be subjected to the measurement screening and then the induction, or may be subjected to the measurement screening after the induction, depending on the test material. The raw material is preferably selected from inbred lines, and preferably from hybrid seeds and comprehensive seeds by inducing and then screening. Or carrying out one-time check measurement after obtaining the inbred line panicle;
(2) and (6) sampling. And taking the fresh-eating-period grains as a detection sample. Sampling in the fresh eating period, breaking off the upper part 1/3-1/2 of each cluster for determination, reserving seeds on the remaining lower clusters and covering a pollination paper bag for protection to prevent water from rotting, and taking 10-15 seeds with regular shapes at the middle part of each cluster.
4. Sample preparation and measurement
Preparing samples and measuring the samples according to the instructions of the instrument. And (4) measuring the central position of the back surface (the surface without the embryo) of the kernel to obtain a measured value. And (4) selecting the fruit ears with the measured values meeting the requirements, and sowing the fruit ears in the next year.
4. Selection system
(1) Selecting according to the intensity value of the fruit peel, selecting the fruit ears with the numerical value smaller than 2.1N, and eliminating the fruit ears with the numerical value larger than the numerical value;
(2) the selected seeds are sowed in single seed, and are sowed according to ear rows or mixed seeds;
(3) and (4) inducing the diploid. Fixing the genotype by utilizing a diploid induction technology to obtain an induction current seed;
(3) sowing the seeds to induce single seeds, and selfing to obtain induced 1-generation fruit clusters. Because the genotype is homozygous and the genotype is consistent with the phenotype, assay screening can begin;
(4) pericarp detection and phenotype identification. And (3) performing peel measurement on the induced 1-generation fruit clusters, sampling in a harvesting period, breaking off the fruit clusters, breaking off the upper parts 1/3-1/2 for measurement, reserving seeds on the remaining lower fruit clusters, and covering a pollination paper bag for protection, so as to prevent water from entering and rotting.
20 grains are taken from each ear close to the middle part of the ear, namely close to the broken part, and the pericarp is measured by a fresh corn pericarp hardness tester. And (4) measuring the sample according to the instruction of the instrument, and measuring the central position of the back surface (the side without the embryo) of the kernel to obtain a measured value. Selecting the ears with the measured value less than 2.1N for selection.
And performing phenotype identification on the appearance of the harvested seeds, wherein the peels at the tops of the seeds have wrinkles, the peels and the endosperm are slightly separated at the corners of the seeds, and the seeds are selected from the normal seeds with normal endosperm, embryo and other parts.
(5) Field identification and crisp and tender reselection of inbred line
Sowing the selected fruit ears, selfing to obtain induced 2-generation ear rows, and performing phenotypic purity identification on the field plants and elimination of undesirable characters. And meanwhile, the selected ear rows are subjected to fruit peel brittleness measurement again, namely, selection. Finally obtaining homozygous ear rows, namely obtaining the pericarp crisp and tender type corn inbred line.
5. Breeding of fresh corn hybrid with crisp and tender pericarp
The pericarp crisp and tender type fresh corn inbred line is used for preparing the pericarp crisp and tender type fresh corn hybrid with parents; or the pericarp crisp and tender type fresh-eating corn inbred line and the non-crisp and tender type fresh-eating corn inbred line are combined into the semi-crisp and tender type fresh-eating corn hybrid, and the palatability of the pericarp is obviously improved.
Example I Breeding of purple waxy corn with crisp and tender pericarp
1. Starting material
Selecting three varieties of purple fresh waxy corn varieties of black waxy corn 600 (Yuju Yu 20170014), black waxy corn 660 (Guju Yu 20200524) and Tian black waxy corn No. 1 (Yuju Yu 2006019), sowing F1 generation 300 plants in an equal amount in a mixed manner, and naturally mixing and pollinating under an isolation condition to harvest.
2. Diploid induction
1000 seeds harvested in the previous generation are sown, and the diploid induction is carried out by using the technology and method of 'a breeding method for inducing parthenogenesis of corn by trifluralin' (patent No. ZL 201110432914.1) invented previously, so that the genotype is fixed. 700 diploid induction seeds were obtained.
3. Crispness and tenderness identification
Sowing and inducing for 1 generation, and selfing to obtain the induced 1 generation individual plant. Sampling in the harvesting period, breaking off the fruit ears, breaking off the upper parts 1/3-1/2 for determination, reserving seeds on the remaining lower fruit ears, covering a pollination paper bag for protection, and preventing water from rotting.
20 grains are taken from each ear close to the middle part of the ear, namely close to the broken part, and the pericarp is measured by a fresh corn pericarp hardness tester. And (4) measuring the sample according to the instruction of the instrument, and measuring the central position of the back surface (the side without the embryo) of the kernel to obtain a measured value. Selecting the ears with the measured value less than 2.1N for selection.
And performing phenotype identification on the appearance of the harvested kernels, wherein the peels at the tops of the kernels are wrinkled, the peels and the endosperm are slightly separated at the corners of the kernels, and the endosperm, the embryo and other parts are normal, so as to obtain 6 ears of the fruit.
4. Field identification and crisp and tender reselection of inbred line
Sowing the selected fruit ears, selfing to obtain induced 2-generation ear rows, and performing phenotypic purity identification on the field plants and elimination of undesirable characters. And meanwhile, the selected ear rows are subjected to fruit peel brittleness measurement again, namely, selection. Finally obtaining 4 rows of homozygous ear rows, namely obtaining 4 sister lines with the serial numbers of C41 and the like of the peel crisp and tender type purple waxy corn inbred line.
5. Breeding of hybrid seeds between sister lines of fresh-eating corn with crisp and tender pericarp
The peel crisp and tender type fresh-eating corn inbred line C41 and the like are used as parents to prepare the peel crisp and tender type fresh-eating corn hybrid, and the palatability of the peel is obviously improved.
6. Effects of the implementation
(1) The hybrid F1 with the parents being crisp and tender peel inbred lines tastes crisp and tender and thin in peel, the peel strength value is about 60 percent of that of the common purple waxy corn, and the hybrid has the mouth feel of melting immediately after being put into the mouth.
(2) Has outstanding taste, is suitable for being processed as a fine product, and has high economic benefit.
Example II Breeding of white waxy corn with crisp and tender pericarp
1. Starting material
Selecting white glutinous fresh-eating corn varieties of Shennuo No. 3 (Liaonong's approved character No. 790) and Wannuo No. 2000 (Guoyouyu 2015032) with good comprehensive properties, sowing the two varieties in an equal amount, mixing and sowing the two varieties into 300 strains of F1, naturally mixing and pollinating the strains under the isolation condition, and reserving the strains.
2. Diploid induction
1000 seeds harvested in the previous generation were sown, and diploid induction was carried out using the "Breeding method for inducing parthenogenesis of maize using trifluralin" (patent No. ZL 201110432914.1) invented previously, to fix the genotype. 600 diploid induction seeds were obtained.
3. Initial selection and identification of brittleness and tenderness
Sowing and inducing for 1 generation, and selfing to obtain the induced 1 generation individual plant. Sampling in the harvesting period, breaking off the fruit ears, breaking off the upper parts 1/3-1/2 for determination, reserving seeds on the remaining lower fruit ears, covering a pollination paper bag for protection, and preventing water from rotting. After sampling, 15 grains are picked at the position of each ear close to the broken part of the ear, and the pericarp is measured by a fresh corn pericarp hardness tester. And (4) measuring the central position of the back surface (the side without the embryo) of the kernel according to the instruction of the instrument to obtain a measured value. Selecting the ear rows with the measured value less than 2.1N for inclusion.
And performing phenotype identification on the appearance of the harvested kernels, wherein the peels at the tops of the kernels are wrinkled, the peels and the endosperm are slightly separated at the corners of the kernels, and the endosperm, the embryo and other parts are normal, so as to obtain 6 ears of the fruit.
4. Inbred line field identification
Sowing the selected fruit clusters, selfing to obtain induced 2-generation ear rows, identifying the phenotype of the field plants, eliminating the poor characters, and selecting and remaining 5 rows of homozygous ear rows to obtain the pericarp crisp and tender white waxy corn inbred line C20 and sister lines.
5. Breeding of fresh corn hybrid with crisp and tender pericarp
Taking a pericarp crisp and tender white waxy corn inbred line C20 and the like as parents to assemble a pericarp crisp and tender fresh corn hybrid; preparing semi-crisp and tender type fresh waxy corn hybrid seeds from the non-crisp and tender type white waxy corn inbred line. The palatability of the peel is significantly improved.
6. Effects of the implementation
(1) The hybrid F1 with the parents being crisp and tender pericarp inbred lines has crisp and tender mouthfeel, thin skin and moderate sweet and glutinous taste.
(2) The F1 fresh taste of the semi-crisp tender type hybrid seed prepared by the crisp and tender peel inbred line and the non-crisp and tender inbred line is also obviously improved, and the peel strength value is about 70 percent of that of the common purple waxy corn.
(3) Has outstanding taste, is suitable for being processed as a fine product, and has high economic benefit.
EXAMPLE III Breeding and utilization of yellow waxy corn with crisp and tender pericarp
1. Starting material
Selecting a yellow glutinous fresh-eating corn variety reclaimed glutinous 1 (national examined corn 2003031) with good comprehensive properties, sowing F1 under isolated conditions, and carrying out natural mixed pollination and seed reservation; and F2 is continuously sown under the isolation condition, and the seeds are naturally mixed and pollinated to obtain F3 seeds.
2. Diploid induction
1000 seeds harvested in the previous generation were sown, and diploid induction was carried out using the "Breeding method for inducing parthenogenesis of maize using trifluralin" (patent No. ZL 201110432914.1) invented previously, to fix the genotype. 650 diploid induction 1-generation seeds were obtained.
3. Crisp and tender identification and screening
Sowing and inducing 1 generation of seeds, and selfing to obtain the ears inducing 1 generation of single plants. Sampling in the fresh eating period, breaking off the fruit ears, breaking off the upper parts 1/3-1/2 for determination, reserving seeds on the remaining lower fruit ears, and sleeving pollination paper bags for protection to prevent water from entering and rotting. 15 grains close to the broken part are picked from each sampled fruit ear, the fruit peel is measured, and the hardness value is measured by a fruit peel hardness meter. And (4) measuring the central position of the fruit skin on the back surface (the side without the embryo) of the kernel according to the instruction of the instrument, and obtaining the measured value. Ears with a measurement value below 2.1N were enrolled.
And performing phenotype identification on the appearance of the harvested seeds, wherein the peels at the tops of the seeds have wrinkles, the peels and the endosperm are slightly separated at the corners of the seeds, and the seeds are selected from the normal seeds with normal endosperm, embryo and other parts. And obtaining the yellow waxy corn ears with crisp and tender pericarp after harvesting.
3. Inbred line field identification
Sowing the selected fruit clusters, selfing to obtain induced 2-generation ear rows, and performing field purity identification. After harvesting, phenotype identification and selection are carried out continuously to obtain pure lines, namely 5 sister lines such as long spike C18 and short spike C20.
5. Breeding of fresh corn hybrid with crisp and tender pericarp
The inbred line C18 of the yellow waxy fresh-eating corn with crisp and tender fruit peel is taken as a female parent, the inbred line C20 of the white waxy fresh-eating corn with crisp and tender fruit peel (bred by the company) is taken as a male parent, and the fruit peel-crisp and tender fresh-eating corn sister line hybrid is prepared.
6. Effects of the implementation
(1) The hybrid F1 has crisp and tender taste, thin skin and mouth feel of melting immediately after being eaten.
(2) The product has outstanding taste, and is suitable for processing as a fine product, and has high economic benefit.
The foregoing is merely a preferred embodiment of the invention, which is intended to be illustrative and not limiting. It will be understood by those skilled in the art that various changes, modifications and equivalents may be made therein without departing from the spirit and scope of the invention as defined in the appended claims.
The principle of the invention is as follows:
(1) according to the phenomenon that the palatability of the pericarp is not necessarily in a negative correlation with the thickness of the pericarp and the rate of the pericarp residue, the possibility of difference of crispness and tenderness of the maize pericarp is presumed, so that a new fresh maize variety with crisp and tender pericarp can be bred; (2) because the maize peel is controlled by polygene and has complex heredity and large difference among different varieties, the maize peel has the diggable heredity potential; (3) the existing fresh-eating corn germplasm created by using a conventional method is heterozygous at the positions of the pericarp characters, namely, the genetic gain has a larger space, so that the breeding selection of the pericarp brittleness is facilitated; (4) the genetic appearance of pericarp differs from that of embryo and endosperm, the appearance of pericarp is not affected by pollen genotype but is solely determined by maternal genotype (sporophyte), and the genotype of the cross is only expressed in the next generation. Therefore, the pericarp character has the problems of inconsistent genotype and phenotype and different generations, and brings trouble to the breeding selection of the pericarp. The diploid induction technology is skillfully used for pericarp breeding, so that the problems of inconsistent pericarp genotype and phenotype and different generations (generations) of the conventional breeding method are solved, and the genotype can be judged through the phenotype.
The diploid induction technique is a "breeding method for inducing parthenogenesis in maize by trifluralin" (201110432914), "a treatment method for inducing parthenogenesis chemically" (ZL 201410405168.0), "an operation method for inducing parthenogenesis in maize chemically" (patent No. ZL 201610337745.6) invented in the present study, and comprises treating filaments with an inducer to induce doubling of the chromosomes of female gametes, thereby directly forming diploid seeds without double fertilization. To distinguish from the haploid induction technique (DH), it is referred to as the diploid induction technique.
The method bypasses the traditional technical route of simply pursuing thin pericarp and low pomace rate, and solves the problems by improving the crispness of the pericarp without considering the pericarp thickness and the pomace rate, in particular to the super-sweet corn (sh 2). The fresh corn hybrid with crisp and tender pericarp is prepared by utilizing the obvious improvement phenomenon of the pericarp palatability shown by crisp and tender, non-crisp and tender and crisp and tender.
First, a new test index, peel strength, is introduced. Namely, the crispness of the peel is represented by the numerical value of the force (unit: cattle) for puncturing the peel by a measuring needle with the diameter of 1.0mm, which is measured by a fresh corn peel hardness tester (ZL 201920602949.7) invented previously. Screening individuals with low strength, namely crisp and tender pericarp through the quantifiable index of the strength value to obtain a fresh-eating corn inbred line with crisp and tender pericarp;
secondly, the corn pericarp is controlled by multiple genes, the pericarp genotype is inconsistent with the embryo and endosperm genotypes, and the separation is asynchronous, namely the pericarp character is not separated among the seeds of the F1 ear, but is separated in the next generation and by taking a single ear as a unit. Aiming at the trouble brought to the conventional breeding by the special genetic behavior of the fruit peel, the 'diploid induction technology' (a breeding method for inducing parthenogenesis of corn by trifluralin, patent technologies such as patent number ZL201110432914.1 and the like) invented before is skillfully used for fruit peel breeding, and the genotype of the selected germplasm is fixed, so that the expression of the genotype and the expression of the phenotype are synchronized, namely, the homozygous genotype with the target character can be accurately obtained through the phenotype. Meanwhile, the homozygous speed is greatly increased, and the crisp and tender peel fresh-eating corn inbred line is obtained.
Thirdly, selection in segregating generations is required in order to fully exploit genetic potential. Based on the fact that the kernel on the F1 ear belongs to F2, namely the female gamete has already started gene recombination separation, F1 can be subjected to induction treatment to homozygously fix various separated genotypes so as to select in the next generation.
The breeding raw material can be any fresh corn such as sweet corn, waxy corn, super sweet corn, etc., hybrid, farmyard or comprehensive species, or any generation such as F1, F2, etc., or semi-finished product of inbred line in the process of inbred line breeding.
By using the technology, the bred inbred line S36 of white waxy fresh corn with crisp and tender pericarp, the inbred line S20 of yellow waxy fresh corn with crisp and tender pericarp, the super sweet corn S26 with crisp and tender pericarp and a plurality of sister lines thereof are bred, and the fresh corn with crisp and tender pericarp prepared by taking the inbred line S20 as a parent is combined. Through phenotype identification, the top pericarp of the pericarp crisp and tender type inbred line seed has wrinkles, the pericarp and the endosperm are slightly separated at the edge of the seed and are easy to damage during friction, and the endosperm, the embryo and other parts are normal.
Claims (4)
1. A breeding method of fresh corn with crisp and tender pericarp is characterized in that: the method comprises the following steps:
(1) original material for breeding
The breeding original material comprises a separation generation of a fresh-eating corn hybrid, the fresh-eating corn hybrid comprises various varieties of sweet corn, super-sweet corn and waxy corn, the separation generation comprises separation generations such as F2 and F3, or semi-finished materials in an inbred line breeding process comprise S1, S2 and S3, or the fresh-eating corn hybrid is obtained by mixed sowing and mixed pollination of a plurality of varieties;
(2) sowing induced single grain
The method for obtaining the induced single particles comprises two methods of screening firstly and then inducing and inducing firstly and then screening; sowing the seeds and inducing single seeds, and selfing to obtain induced 1-generation fruit clusters;
the method for screening firstly and inducing secondly comprises the following steps:
the raw material is an inbred line, and the breeding raw material is subjected to brittleness detection and screening to obtain crisp and tender corn seeds; performing single-seed sowing on the crisp and tender corn seeds, performing induction line selection on the sowed ear rows or single plants according to ear rows or mixed sowing, and obtaining induced single seeds through diploid induction;
the method for inducing firstly and screening secondly comprises the following steps:
the raw material is hybrid seeds and comprehensive seeds; obtaining current-generation induced seeds through diploid induction; performing brittleness detection screening to obtain induced single grains;
the brittleness detection screening method comprises the following steps: taking fresh corn kernels as a detection sample, measuring the crispness and tenderness by using the force (unit: cattle) for puncturing the pericarp by a measuring needle with the diameter of 1.0mm, and determining the value to be less than 2.1N;
(3) pericarp detection and phenotype identification:
performing brittleness detection on the induced 1-generation fruit cluster, wherein the genotype of the induced 1-generation fruit cluster is homozygous, and the genotype is consistent with the phenotype, so that a pericarp crisp and tender type fresh-eating corn inbred line is obtained;
the brittleness detection method is the same as the brittleness detection screening method in the step (2); the fruit peel strength value of the fresh-eating period fruit peel of the crisp and tender type fresh-eating corn inbred line is less than 2.1N, the fruit peel at the top end of mature seed grains has wrinkles, and endosperm, embryos and other parts are normal.
2. The method for breeding the fresh-eating corn with crisp and tender pericarp as claimed in claim 1, which is characterized in that: the brittleness detection method adopts a corn peel hardness tester for testing, and the corn peel hardness tester is a fresh corn peel hardness tester (the patent number is ZL 201920602949.7); sampling in the fresh eating period, breaking off the upper parts 1/3-1/2 of the ears for determination, reserving seeds on the rest ears, covering pollination paper bags for protection, preventing water from entering and rotting, and taking 10-15 grains with regular shapes at the middle part of each ear.
3. The method for breeding the fresh-eating corn with crisp and tender pericarp as claimed in claim 1, which is characterized in that: and (3) performing induction 1-generation selfing to obtain induction 2-generation ear rows, performing field plant phenotype purity identification and elimination on poor characters, performing fruit peel brittleness determination on the selected ear rows again, namely checking, and finally obtaining homozygous ear rows, namely obtaining the fruit peel crisp and tender type corn inbred line.
4. The method for breeding the fresh-eating corn with crisp and tender pericarp as claimed in claim 1, which is characterized in that: the pericarp crisp and tender type fresh corn inbred line is used for preparing crisp and tender pericarp hybrids, and the specific method comprises the following steps: the pericarp crisp and tender type fresh-eating corn inbred line is multiplied by the pericarp crisp and tender type fresh-eating corn inbred line to prepare the crisp and tender type pericarp fresh-eating corn hybrid, or the pericarp crisp and tender type fresh-eating corn inbred line is multiplied by the non-pericarp crisp and tender type fresh-eating corn inbred line to prepare the semi-crisp and tender type fresh-eating corn hybrid.
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