CN111165344B - Method for rapidly creating sweet corn inbred line rich in anthocyanin - Google Patents

Method for rapidly creating sweet corn inbred line rich in anthocyanin Download PDF

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CN111165344B
CN111165344B CN202010120607.9A CN202010120607A CN111165344B CN 111165344 B CN111165344 B CN 111165344B CN 202010120607 A CN202010120607 A CN 202010120607A CN 111165344 B CN111165344 B CN 111165344B
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anthocyanin
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李春艳
李高科
胡建广
李武
卢文佳
文天祥
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Shenzhen A Corn Technology Co ltd
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CROP Research Institute of Guangdong Academy of Agricultural Sciences
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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
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • 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
    • 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
    • A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
    • A01H1/08—Methods for producing changes in chromosome number

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Abstract

The invention belongs to the technical field of crop breeding, and particularly relates to a method for quickly creating a sweet corn inbred line rich in anthocyanin, which comprises the following steps: 1) selecting a colorless corn inbred line to be hybridized with the corn rich in anthocyanin to obtain F1Seed generation; 2) planting F1Generation of seed, F1Selfing the plant to obtain F2Replacing seeds, and selecting dark seeds; 3) hybridizing the dark seeds with a high-oil induction line to obtain F2ySeeds; 4) selecting haploid seeds by adopting a nuclear magnetic resonance sorting system, and doubling the haploid seeds after planting the haploid seeds to obtain homozygous doubled haploid seeds DH; 5) and (3) selecting the seeds which are rich in anthocyanin and shrink from the DH seeds to obtain the required sweet corn inbred line rich in anthocyanin. By using the haploid doubling method, the breeding process is shortened, and the homozygous anthocyanin-rich maize inbred line can be obtained through 4 generations in the breeding process to obtain the double haploid pure line.

Description

Method for rapidly creating sweet corn inbred line rich in anthocyanin
Technical Field
The invention belongs to the technical field of crop breeding, and particularly relates to a method for quickly creating a sweet corn inbred line rich in anthocyanin.
Background
The anthocyanin is a soluble flavonoid substance existing in plants in the nature, is a main color substance in the plants, the most main physiological activity function of the anthocyanin in the plants is free radical scavenging capacity and antioxidant capacity, researches prove that the anthocyanin is the most effective antioxidant found by human at present and is also the most effective free radical scavenger, the antioxidant capacity of the anthocyanin is 50 times higher than that of vitamin E and 20 times higher than that of vitamin C, the bioavailability of the anthocyanin to human bodies is one hundred percent, and the anthocyanin can be detected in blood 20min after being taken. Meanwhile, researches show that other activity mechanisms of anthocyanin are beneficial to human health, and the anthocyanin and the bioflavonoid are rich in anthocyanin mixture, so that the anthocyanin and the bioflavonoid have the functions of preventing DNA cracking, improving estrogenic activity, inhibiting enzyme, promoting production factors and the like, regulating immune response and resisting inflammatory activity.
The existing black corn rich in anthocyanin, also called purple corn, is originally produced in south America, and gradually becomes one of favorite health foods along with the pursuit of people for quality of life, but the black corn is still not very popular in the market, so that the production mode of the black corn is slightly difficult and the growth cycle is longer.
The prior patent CN107396829A discloses a method for breeding a pericarp anthocyanin synthesized fresh-eating corn inbred line, which adopts the hybridization of a corn inbred line with black or purple pericarp and a corn inbred line with colorless pericarp, and finally selects a full-black or full-purple corn inbred line in an n-generation inbred mode;
also, patent CN107347634B discloses a method for breeding a synthetic fresh sweet corn inbred line of anthocyanin based on leaf tongue color, which adopts a traditional hybridization method to accelerate the breeding process according to leaf tongue color breeding, wherein the inbred algebra still needs to reach 5-7 generations;
and a patent CN108293873A discloses a breeding method of black sweet waxy corn, which adopts a traditional selfing mode, screens out purple black waxy grains by a minimally invasive single-grain iodine solution identification method, selects an excellent corn selfing line to carry out selfing continuously, but the subsequent selfing still needs to carry out 7-8 generations for breeding.
Disclosure of Invention
Aiming at the problems, in the prior art, the period for producing the black corn rich in anthocyanin is long, the period for 5-10 generations of inbred lines needs to be waited, the cultivation process is slightly complex, the time cost, the labor cost and the like are wasted, and the anthocyanin content of the variety after multi-generation inbreeding is influenced. Therefore, the invention provides a method for quickly creating the sweet corn inbred line rich in anthocyanin, and provides the sweet corn inbred line which can quickly produce high-quality sweet corn inbred line rich in anthocyanin with high yield.
The technical content of the invention is as follows:
the invention provides a method for rapidly creating a sweet corn inbred line rich in anthocyanin, which comprises the following steps:
1) selecting a colorless corn inbred line to be hybridized with the corn rich in anthocyanin to obtain F1Seed generation;
2) planting F1Generation of seed, F1Selfing the plant to obtain F2Replacing seeds, and selecting dark seeds;
3) hybridizing the dark seeds with a high-oil induction line to obtain F2ySeeds;
4) selecting haploid seeds by adopting a nuclear magnetic resonance sorting system, and doubling the haploid seeds after planting the haploid seeds to obtain homozygous doubled haploid seeds DH;
5) and (3) selecting the seeds which are rich in anthocyanin and shrink from the DH seeds to obtain the required sweet corn inbred line rich in anthocyanin.
Further, the colorless corn of step 1) comprises yellow or white sweet corn;
further, the maize rich in anthocyanin in the step 1) comprises the maize with red peel, purple peel or black peel;
further, the maize rich in anthocyanin in the step 1) comprises sweet maize or waxy maize or feeding maize, and the maize rich in anthocyanin can be used as the hybrid female parent in the step 1);
further, the haploid doubling in step 4) comprises haploid doubling with colchicine.
The invention has the following beneficial effects:
according to the method for rapidly creating the anthocyanin-rich sweet corn inbred line, the breeding process is shortened by utilizing a haploid doubling method, the homozygous anthocyanin-rich corn inbred line can be obtained through 4 generations in the breeding process, the obtained double haploid pure line (DH line) is 100% homozygous in theory, compared with the technology which can be completed through 8-10 generations in the prior art and has no high purity of the DH line, the technology can realize rapid obtaining of the high-content anthocyanin-rich corn inbred line;
for the selection of the haploid seeds, the traditional method is manual selection, the selection is carried out according to the fact that the top end of corn kernels has the color of an induction line, and embryos have no color, but the top end of the corn kernels rich in anthocyanin has the color, the color is overlapped with the marked color of the induction line, the judgment is difficult by naked eyes, and the manual selection cannot be completed.
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FIG. 1 shows a selection of colorless sweet corn and anthocyanin-rich sweet corn;
FIG. 2 is an anthocyanin-rich sweet maize inbred line.
Detailed Description
The present invention is described in further detail in the following description of specific embodiments and the accompanying drawings, it is to be understood that these embodiments are merely illustrative of the present invention and are not intended to limit the scope of the invention, which is defined by the appended claims, and modifications thereof by those skilled in the art after reading this disclosure that are equivalent to the above described embodiments.
All the raw materials and reagents of the invention are conventional market raw materials and reagents unless otherwise specified.
Example 1
A method for rapidly creating a sweet corn inbred line rich in anthocyanin comprises the following steps:
1) selecting a high-quality yellow sweet corn inbred line and the anthocyaninCrossing black corn of the seed to obtain F1Seed generation;
2) planting F1Generation of seed, F1Selfing the plant to obtain F2Replacing seeds, and selecting black seeds;
3) hybridizing black seeds with a high-oil induction line to obtain F2ySeeds;
4) because the induction frenulum is marked and according to the difference of oil content, haploid seeds are selected by adopting an OnlinemR20-015V-S type nuclear magnetic resonance sorting system, and the haploid seeds planted are doubled by the haploid through colchicine to obtain homozygous double haploid seeds DH;
5) selecting black and shrunken seeds from DH seeds to obtain the required black sweet corn inbred line rich in anthocyanin;
the obtained black sweet corn inbred line can be used as a parent material for cultivating new black sweet corn varieties rich in cyanine. The black sweet corn inbred line obtained by the method is hybridized with a yellow or white sweet corn inbred line to obtain a new black sweet corn variety rich in anthocyanin.
Example 2
A method for rapidly creating a sweet corn inbred line rich in anthocyanin comprises the following steps:
1) selecting a high-quality yellow sweet corn inbred line to be hybridized with the purple corn rich in anthocyanin to obtain F1Seed generation;
2) planting F1Generation of seed, F1Selfing the plant to obtain F2Carrying out seed generation, namely selecting purple seeds;
3) hybridizing the purple seeds with a high-oil induction line to obtain F2ySeeds;
4) because the induction frenulum is marked and according to the difference of oil content, haploid seeds are selected by adopting an OnlinemR20-015V-S type nuclear magnetic resonance sorting system, and the haploid seeds planted are doubled by the haploid through colchicine to obtain homozygous double haploid seeds DH;
5) selecting purple and shrunken seeds from DH seeds to obtain the required purple sweet corn inbred line rich in anthocyanin;
the obtained purple sweet corn inbred line can be used as a parent material for cultivating new purple sweet corn varieties rich in cyanine. The purple sweet corn inbred line obtained by the method is hybridized with a yellow or white sweet corn inbred line to obtain a new purple sweet corn variety rich in anthocyanin.
Example 3
A method for rapidly creating a sweet corn inbred line rich in anthocyanin comprises the following steps:
1) selecting a high-quality white sweet corn inbred line to be hybridized with the red corn rich in anthocyanin to obtain F1Seed generation;
2) planting F1Generation of seed, F1Selfing the plant to obtain F2Replacing seeds, and selecting red seeds;
3) hybridizing the red seeds with a high-oil induction line to obtain F2ySeeds;
4) because the induction frenulum is marked and according to the difference of oil content, haploid seeds are selected by adopting an OnlinemR20-015V-S type nuclear magnetic resonance sorting system, and the haploid seeds planted are doubled by the haploid through colchicine to obtain homozygous double haploid seeds DH;
5) selecting red and shrunken seeds from DH seeds to obtain the required red sweet corn inbred line rich in anthocyanin;
the obtained red sweet corn inbred line can be used as a parent material for cultivating new red sweet corn varieties rich in cyanine. The red sweet corn inbred line obtained by the method is hybridized with a yellow or white sweet corn inbred line to obtain a new red sweet corn variety rich in anthocyanin.
Comparative example 1
The method comprises the following steps of (1) culturing the anthocyanin-rich maize inbred line by adopting a traditional method:
1) selecting a high-quality yellow sweet corn inbred line to be hybridized with the purple corn rich in anthocyanin to obtain F1Seed generation;
2) f is to be1Reserving seeds for self-breeding to obtain F2Generation;
3) f is to be2Sowing the seeds in different generations, selecting the phenotypic characters of specific plant type, leaf type and disease resistanceSelfing the excellent single plant to obtain F3Generation;
4) f is to be3Planting according to ear rows, selecting single plants with purple full ear fruit skins and excellent phenotypic characters such as plant type, leaf type, disease resistance and the like, selfing and reserving single ears to obtain F4Generation;
5) f is to be4Planting according to ear rows, selecting single plants with purple full ear fruit skins and excellent phenotypic characters such as plant type, leaf type, disease resistance and the like, selfing and reserving single ears to obtain F5Generation;
6) f is to be5Planting according to ear rows, selecting single plants with purple full ear fruit skins and excellent phenotypic characters such as plant type, leaf type, disease resistance and the like, selfing and reserving single ears to obtain F6Generation;
7) f is to be6Planting according to ear rows, selecting single plants with purple full ear fruit skins and excellent phenotypic characters such as plant type, leaf type, disease resistance and the like, selfing and reserving single ears to obtain F7Generation;
8) f is to be7Planting according to ear rows, selecting single plants with purple full ear fruit skins and excellent phenotypic characters such as plant type, leaf type, disease resistance and the like, selfing and reserving single ears to obtain F8Generation;
9) f is to be8And planting according to ear rows, selecting a single plant with purple full ear fruit skin and excellent phenotypic characters such as plant type, leaf type, disease resistance and the like for selfing and reserving seeds, and obtaining a purple maize selfing line rich in anthocyanin.
The anthocyanin-rich corn of step 1) in examples 1-3 and comparative example 1 above includes sweet corn, waxy corn, or forage corn.
The maize inbred lines obtained in examples 1 to 3 and comparative example 1 were subjected to anthocyanin content measurement by liquid chromatography, and the measurement results were as follows:
TABLE 1 anthocyanin content of maize inbred lines
Figure BDA0002392852640000071
The above examples all show that the method for cultivating the anthocyanin-rich sweet corn inbred line only needs 4 generations, and the obtained anthocyanin-rich corn inbred line of the double haploid pure line greatly improves the efficiency of cultivating the anthocyanin-rich corn inbred line and saves manpower, material resources and time cost compared with the prior art.
As shown in figure 1, the selected colorless corn and the anthocyanin-rich corn are shown, and figure 2 is the anthocyanin-rich shrunken sweet corn.

Claims (2)

1. A method for rapidly creating a sweet corn inbred line rich in anthocyanin is characterized by comprising the following steps:
1) selecting a colorless corn inbred line to be hybridized with the corn rich in anthocyanin to obtain F1Seed generation;
the colorless corn comprises yellow or white sweet corn;
the maize rich in anthocyanin comprises red peel, purple peel or black peel maize;
the maize rich in anthocyanin comprises sweet maize or waxy maize or feeding maize;
2) planting F1Generation of seed, F1Selfing the plant to obtain F2Replacing seeds, and selecting dark seeds;
the dark seeds comprise red peel, purple peel or black peel seeds;
3) hybridizing the dark seeds with a high-oil induction line to obtain F2ySeeds;
4) selecting haploid seeds by adopting a nuclear magnetic resonance sorting system, and doubling the haploid seeds after planting the haploid seeds to obtain homozygous doubled haploid seeds DH;
5) and (3) selecting the seeds which are rich in anthocyanin and shrink from the DH seeds to obtain the required sweet corn inbred line rich in anthocyanin.
2. The method of anthocyanin-rich sweet corn inbred line of claim 1, wherein the haploid doubling in step 4) comprises haploid doubling with colchicine.
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101377481A (en) * 2008-10-06 2009-03-04 中国农业大学 Method for identifying corn parthenogenesis haploidy
CN104521742A (en) * 2014-12-26 2015-04-22 广东省农业科学院作物研究所 Method for rapidly creating sweet-waxy recessive maize inbred line
CN107347634A (en) * 2017-09-13 2017-11-17 安徽省农业科学院烟草研究所 A kind of method based on tip of a leaf color assist-breeding pericarp anthocyanidin synthesis type fresh edible maize self-mating system

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101377481A (en) * 2008-10-06 2009-03-04 中国农业大学 Method for identifying corn parthenogenesis haploidy
CN104521742A (en) * 2014-12-26 2015-04-22 广东省农业科学院作物研究所 Method for rapidly creating sweet-waxy recessive maize inbred line
CN107347634A (en) * 2017-09-13 2017-11-17 安徽省农业科学院烟草研究所 A kind of method based on tip of a leaf color assist-breeding pericarp anthocyanidin synthesis type fresh edible maize self-mating system

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
玉米单倍体的诱导及其化学加倍方法;魏俊杰等;《安徽农业科学》;20110110;第39卷(第2期);全文 *
玉米单倍体育种技术研究及应用;陈绍江;《2019年中国作物学会学术年会论文摘要集》;20191027;第7页第1-5行，第11-18行 *
玉米高油型诱导系CHOI1和CHOI3的单倍体鉴定效率评价;董昕等;《中国农业大学学报》;20190615;第24卷(第6期);第2页左栏第1段第1-6行、第2段，第2页右栏第1段第1-5行、第3段，第3页第1.3节，第9页左栏第4节 结论 *

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Address after: 518000 Xinghe WORLDA801-02, No.1 Yabao Road, Nankeng Community, Bantian Street, Longgang District, Shenzhen, Guangdong

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Patentee before: CROP Research Institute GUANGDONG ACADEMY OF AGRICULTURAL SCIENCES