CN111972277B - Method for breeding black wheat germplasm with different genetic backgrounds by utilizing blue dwarf male sterile and purple wheat - Google Patents
Method for breeding black wheat germplasm with different genetic backgrounds by utilizing blue dwarf male sterile and purple wheat Download PDFInfo
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- CN111972277B CN111972277B CN202010800466.5A CN202010800466A CN111972277B CN 111972277 B CN111972277 B CN 111972277B CN 202010800466 A CN202010800466 A CN 202010800466A CN 111972277 B CN111972277 B CN 111972277B
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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
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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
- A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
- A01H1/08—Methods for producing changes in chromosome number
Abstract
The invention belongs to the technical field of wheat breeding, and particularly relates to a method for breeding different genetic backgrounds of rye germplasm by using blue dwarf male sterile and purple wheat. Continuously pollinating blue dwarf-male-sterile backcross by purple wheat, transforming the blue dwarf-male-sterile purple wheat into blue dwarf-male-sterile purple wheat, selecting black grain fertile plant material containing blue grains and purple grain genes separated by breaking the linkage from the transformed blue dwarf-male-sterile purple wheat, radiating the black grains and the blue dwarf-male-sterile purple wheat, pollinating the processed blue dwarf-male-sterile purple wheat by using the processed black grains, selecting homozygous whole black wheat strains, and confirming the genetic background type of the germplasm of the bred blue wheat through root cap tissue tabletting and dyeing microscopic examination. The method for breeding the black wheat is reliable and efficient.
Description
Technical Field
The invention belongs to the technical field of wheat breeding, and particularly relates to a method for breeding different genetic backgrounds of rye germplasm by utilizing blue dwarf male sterile and purple wheat.
Background
Wheat is a main grain crop of people in China, and is the most main staple food of people in northern areas of China, the 'recessive hunger' caused by unbalanced nutrients can be solved through the staple food, the hidden health hazards are relieved, the method is promoted by the world health organization and the international food and rink organization, and the method for solving the problems of insufficient nutrients such as trace elements and vitamins and unbalance through biological strengthening is the same.
The blue wheat contains anthocyanin with an anti-oxidation function in an aleurone layer and an endosperm, can enrich microelements such as zinc, iron, chromium, manganese, selenium and the like which are beneficial to a human body from soil, has a strong health-care effect, and simultaneously, the ground flour has good taste. The purple wheat and the blue wheat have similar or same health care efficacy, but pigments are concentrated in seed skins and are not easy to utilize in the process of grinding, and if the whole wheat flour is eaten, the whole wheat flour has slightly poor taste, but has the same nutrition and health care efficacy as the blue wheat. The rye is a wheat germplasm material containing purple pericarp genes and blue aleurone layer genes. The rye whole wheat flour has higher and better nutritive value and has important significance for special groups of patients with diabetes, cardiovascular diseases, obesity and the like.
However, blue wheat and purple wheat have very unique characteristics in botany and genetics. From a plant perspective, wheat seeds are actually a fruit called caryopsis, which develops from the whole ovary, the outermost "bran" of the seeds is the pericarp and the pericarp that are developed from the ovary wall and the integument are healed together and are mixed by us as the "seed coat", the seed coat is the aleurone layer and the inner mass of endosperm. Purple pigment of purple wheat exists in pericarp and develops from the ovary wall of a mother body, so that the phenomenon of maternal heredity exists on heredity, the phenomenon of next generation exists on the separation of filial generation, the character separation and recombination of the purple wheat is characterized in that the character separation and recombination occur in F3 generation, and the purple wheat is a result of the gene action of purple pericarp nucleus; the blue wheat is actually a blue aleurone layer gene, the aleurone layer belongs to the outermost layer cell of triploid endosperm and a part of endosperm, and the current endosperm is directly felt in the aspect of property expression. The characteristics of botany and genetics necessarily reflect the uniqueness of the breeding method and technology when the black wheat with blue aleurone layer and purple pericarp gene is bred simultaneously.
The blue dwarf male sterile line is a wheat sterile line, belongs to a dominant genic male sterile line of a blue grain monomer type, and has three dominant characters of blue grain (4D/4E), dwarf (Rht 10) and anther abortion (Ms 2) which are interlocked on a translocation addition line 4D # chromosome monomer.
Blue grains are dominant and white grains are invisible in blue dwarf male-sterile, and in the process of backcrossing and blue dwarf male-sterile by using a white grain variety, different numbers of blue grain fertile plants are often separated, which causes troubles for seed production work of wheat heterosis utilization.
Researches of scholars consider that purple wheat has two pairs of independent dominant complementary gene controls, purple grains are white grains in wheat trait inheritance, and purple is dominant; however, purple appears invisible to blue, while blue is dominant. The wheat grains containing blue and purple double genes are remarkably deepened, and are called black grains. When the purple seed variety is pollinated by blue dwarf male sterile, the blue seed is expressed as a sterile line, the purple seed is a fertile plant, and the black seed containing blue and purple double genes can be separated by breaking the linkage in the blue dwarf male sterile. Although blue grains are dominant to blue grains in purple grains, the blue grains are not easy to be transferred as white grains are transferred in a backcross transfer process by utilizing blue dwarf-male-sterile, the color difference between blue, purple and black is not obvious, the blue, purple and black colors are required to be identified carefully, the blue dwarf-male-sterile-breeding method cannot be used for large-scale production and utilization of hybrid wheat seed production and other fields, but is used for small-scale breeding, and the blue dwarf-male-sterile-breeding method still shows the unique advantages of convenience and high efficiency by taking the blue dwarf-male-sterile as a tool carrier for breeding the black wheat. The approaches of purple wheat hybridization, backcross transformation and separation of rye are more convenient and practical compared with other methods and other tool carriers. Until now, no one has been breeding the black wheat containing blue and purple genes by utilizing blue dwarf male sterile, and no one has been breeding the black wheat with different genetic types by utilizing black grains containing blue and purple genes as male parents to pollinate the blue dwarf male sterile.
Disclosure of Invention
The invention aims to overcome the defects of the background technology and provide a method for breeding different genetic background black wheat germplasms by utilizing blue dwarf-male-sterile and purple wheat.
The invention is realized by the following technical scheme:
a method for breeding black wheat germplasm with different genetic backgrounds by utilizing blue dwarf-male-sterile and purple wheat includes utilizing purple wheat to continuously backcross and pollinate blue dwarf-male-sterile wheat germplasm belonging to 4D/4E blue grain translocation line monomer, transferring into blue dwarf-male-sterile purple wheat, selecting black grains containing blue grains and purple grain genes separated by breaking linkage from the transferred blue dwarf-male-sterile purple wheat, carrying out radiation treatment on the transferred blue dwarf-male-sterile purple wheat and the selected black grains, and pollinating the radiation-treated blue dwarf-male-sterile purple wheat by the radiation-treated black grains so as to select and breed the black wheat germplasm.
The radiation treatment adopts 60 C O -r-rays.
The described 60 C O The gamma ray dose is 50-60% lethal dose.
The bred rye germplasm comprises a 4D/4E translocation twin-hull rye germplasm and a 4D/4E translocation substitution line rye germplasm.
The 4D/4E translocation twin-block rye germplasm and the 4D/4E translocation replacement line rye germplasm both utilize root tip tabletting and observe the chromosome number through microscopic examination to confirm the genetic background type of the bred rye.
The method for breeding the black-wheat germ plasm which is the near isogenic line black-wheat strain comprises the following steps: selecting black-grain fertile plants separated by breaking the linkage from the blue dwarf-male-sterile purple wheat which is stably transferred, further breeding whole black-grain selected plants, and then mixing all the plants to obtain the near-isogenic black-wheat strain.
The purple wheat is Shannon purple wheat No. 1, and the blue dwarf male-sterile wheat germplasm is blue dwarf male-sterile Luan 02-1.
The purple wheat is Loxon 1, and the blue dwarf male-sterile wheat germplasm is blue dwarf male-sterile black horse 1.
The invention has the beneficial effects that:
the invention discloses a method for breeding black wheat germplasm by using blue dwarf-male-sterile wheat and purple wheat, and particularly discloses a method for breeding blue purple wheat (black wheat) germplasm, the seed coat of which contains a purple pigment gene and also contains an aleurone layer blue gene, by using blue dwarf-male-sterile germplasm of a 4D/4E blue grain translocation series monomer. The black wheat contains purple pericarp genes and blue aleurone layer genes, has high content of anthocyanin, is rich in beneficial trace elements such as zinc, iron, manganese, chromium, selenium and the like, has high health care nutritive value, is transformed into blue dwarf-male-sterile purple wheat by continuously pollinating blue dwarf-male-sterile with purple wheat, selects black grain fertile plant materials containing blue grains and purple grain genes separated by breaking linkage from the transformed blue dwarf-male-sterile purple wheat, performs radiation treatment on the black grains and the blue dwarf-male-sterile purple wheat, pollinates with the treated black grains as the treated blue dwarf-male-sterile purple wheat, and then selects a homozygous whole black wheat strain.
4D/4E translocation Lonicera japonica germplasm Am-HF1 and Am-HF3 of 44 chromosomes are bred by the method; and 4D/4E translocation substitution line Umbelliferae germplasm HF-Am2, Am-HF3 with 42 chromosomes. The method for breeding the rye is reliable and efficient.
Compared with the prior art, the method has the unique advantages of convenience, high efficiency and the like by taking the blue dwarf male sterile as a tool carrier for breeding the rye. Compared with other existing methods and other tool carriers, the method for hybridizing, backcrossing and transforming purple wheat and separating the black wheat is more convenient and practical.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The blue dwarf-male-sterile wheat is backcrossed and pollinated by purple wheat continuously, the blue dwarf-male-sterile purple wheat is transferred, black grain materials containing blue grain and purple grain genes separated by breaking the linkage are selected from the transferred blue dwarf-male-sterile purple wheat, and the black grain materials are used 60 C O -r ray (50-60% lethal dose) is used for treating the black grains and the blue dwarf male sterile purple wheat to increase the chance of breaking linkage, changing linkage position and increasing character exchange probability, which is beneficial to forming black wheat germplasm materials with different genetic backgrounds in later period, then the radiation-treated black grains are used for pollinating the radiation-treated blue dwarf male sterile purple wheat, black wheat fertile plants are selected from the offspring group, further, homozygous whole black wheat strains with different genetic backgrounds are selected, and the root tip cell tabletting lens is used for carrying out tablettingThe genetic type of the rye is detected and confirmed.
Example 1:
the method comprises the following steps of breeding black wheat germplasm including HF-Am1 black wheat strain and HF-Am2 black wheat strain by using blue dwarf patrinia Luan 02-1 and Shannon purple wheat No. 1 as materials, and specifically comprises the following steps:
and (3) breeding blue dwarf male sterile Shannon purple wheat No. 1: the method comprises the steps of sowing blue grains (blue dwarf-male-sterile) sorted from a blue dwarf-male-sterile population Luan 02-1 in autumn, planting 2 lines, 2 meters in line length, 25 grains/meter and 20 centimeters in line spacing, sowing 1 line of pollinated male parent Shannong purple wheat No. 1 on one side, sowing male parent Shannong purple wheat No. 1 on the other side at the second period before overwintering, sowing 100 grains/meter, occupying 0.8 meter of land for sowing, and respectively reserving 1 meter of small walkways around the sowing belt for experimental observation. Before heading, bamboo-wood piles, iron wires and plastic woven cloth are used for building an isolation area of the isolation wall. The method is characterized in that various separated anther fertile plants are pulled out in time, natural pollination is carried out in the glume pollination period, or the plants are stirred by a small bamboo rod to be artificially and auxiliarily pollinated, when the plants are mature, the blue dwarf male sterile of purple wheat pollen is harvested, grains formed by the blue dwarf male sterile are still blue and white due to the fact that purple is inherited as a parent, and the sorted blue grains are sowed in autumn for later use.
The autumn sowing is in the same field layout as the last year, the blue grains sorted in 2 rows are sown in a parent planting mode, shannong purple wheat No. 1 sown in two sides in stages is used as a backcross male parent, a separation wall is built, natural pollination is carried out, after the generation, the colors of the seeds on the blue dwarf male sterile plants are blue grains and purple grains, and the blue grains are sorted and continue to be sown in the autumn.
Repeating the field planting and indoor blue grain sorting work with the purple grain wheat Shannon purple wheat No. 1 as the backcross male parent and the blue grain blue dwarf male sterile line as the pollination carrier year after year until the backcross is stabilized to be the blue dwarf male sterile Shannon purple wheat No. 1, and properly expanding the population in the last year of the next link breeding to prepare enough seeds for the next link.
Selecting black fertile plants: sowing blue-grain blue dwarf-male-sterile purple wheat sorted from blue dwarf-male-sterile Shannong purple wheat No. 1 backcross populations by using a sowing building with a seed sowing plate in autumn, sowing 6 rows with the row spacing of 20 cm and the row length of 20 m, selecting a seed sowing plate of 5 kg/mu, wherein the population scale is about 5000 grains,the male parent Shannon purple wheat No. 1 is sown for pollination in 1 row on one side, the distance between the male parent and blue dwarf male sterile purple wheat is 30 cm and 30 grains/meter, the male parent Shannon purple wheat No. 1 and 100 grains/meter are sown in the second period before overwintering on the other side, and small walkways of 1 meter are respectively reserved around a sowing plot for experimental observation. Before heading, bamboo-wood piles, iron wires and plastic woven cloth are used for building isolation walls to build isolation areas. After heading, the fertile plants with anther, namely fertile black-grain plants, are searched in blue dwarf male-sterile Shannon purple wheat No. 1 and separated out through breaking chain, the whole plants are bagged and branded, the mature period is waited, black grains and purple grains are sorted according to the harvest of single plants, the black grains sorted from each selected plant are mixed (if the seed quantity is small, propagation can be expanded and breeding can be carried out for one year), the backcross groups of blue dwarf male-sterile Shannon purple wheat No. 1 are synchronously harvested, blue grain blue dwarf male-sterile wheat is sorted, the sorted blue grain blue dwarf male-sterile purple wheat and the selected black-grain fertile plant grains are adopted together before autumn sowing 60 C O -gamma-ray radiation treatment with 50% -60% lethal dose for sowing.
The radiation treatment of the black-grain fertile plants is to pollinate blue dwarf-male-sterile Shannon purple wheat No. 1: sowing 6-row channels of sowing building with seed discharging plates in autumn 60 C O The method comprises the steps of-irradiating blue dwarf-male-sterile Shannon purple wheat No. 1 with 50-60% of gamma ray lethal dose, wherein the row length is 10 m, using a seed sowing plate (the size of more than 2000 colonies, about 800 survives), planting black grains which are selected, sorted and subjected to radiation treatment on male parents at two sides, 30 grains/m, building an isolation wall by using bamboo piles, iron wires and plastic woven cloth before heading to construct an isolation region, naturally pollinating the blue dwarf-male-sterile Shannon purple wheat No. 1 with the black grains fertile plants in the isolation region, or stirring artificial auxiliary pollination by using a small bamboo rod, harvesting the blue dwarf-male-sterile Shannon purple wheat No. 1 with the black grains pollen when the blue dwarf-male-sterile Shannon purple wheat is mature, mixing, harvesting, mixing, threshing, sorting the black grains and purple grains, and establishing a seed selection garden after autumn sowing.
Selecting a whole black-grain plant: the black grains are thinly sown and sorted by a sowing building (or a plot sowing building) with a seed arrangement disc in autumn, the sowing amount is 3-5 kg, and an isolation area is built before heading to prevent pollution to other blue dwarf-male-sterile materials. The seeds can separate partial blue dwarf male sterile plants in the field, most of the blue dwarf male sterile plants are black fertile plants, full black plants exist in the black fertile plants, and single plants with two grain colors of black grains and purple grains of the same plant also exist, so that the purpose of establishing the seed selection garden is to breed the full black plants. And distinguishing the whole black plants one by one in the mature period, pulling out the whole black plants, and selecting the plants with inconsistent grain colors and relative expression deviation by selecting the seeds. And in consideration of the characteristic that purple characters show maternal inheritance, the seeds sorted by all selected individual plants are mixed and planted and selected for one generation according to the mode of the previous generation so as to ensure that the selected individual plants are of a real gene recombination type.
Breeding and stabilizing the rye strain: after selecting the whole black grain single plants of two generations, selecting the whole black grain selected plants according to the single plant sowing in autumn, sowing 3 rows of each single plant, sowing the row spacing of 20 cm, 30 grains/meter and 3 meters of row length, carrying out field investigation and recording, observing the stable consistency condition of each strain, finally threshing according to the single harvest of the strain and metering the yield of the whole area. And (4) re-establishing ears and purifying the unstable rye strains with excellent comprehensive performance. And (3) carrying out yield comparison on the selected whole black-kernel strains, carrying out seed indoor germination, carrying out root tip tabletting, observing the number of chromosomes by microscopic examination, confirming that a good strain HF-Am1 with 44 chromosomes is 4D/4E translocation two-body black-wheat germ plasm, and simultaneously breeding 4D/4E translocation substitution line black-wheat germ plasm HF-Am2 with 42 chromosomes.
And all-UK-grain fertile strains selected from basically stable blue dwarf-male-sterile Shannon purple wheat No. 1 are mixed and bred to form the near isogenic line UK wheat strain of the Shannon purple wheat No. 1, and the order production of UK wheat is carried out, which is a breeding method of the near isogenic line UK wheat.
Example 2:
the blue dwarf-male-sterile black wheat No. 1 and the complex No. 1 purple wheat are used as materials to breed black wheat strains, namely HF-Am3 and HF-Am4 black wheat strains.
The same approach and method as in example 1 are adopted, the self-bred blue dwarf-rancidity black horse No. 1 and purple wheat of the complex No. 1 are transformed into blue dwarf-rancidity complex No. 1, then black grain fertile plants are bred from the blue dwarf-rancidity complex No. 1, then the selected black grains and the blue grains sorted from the blue dwarf-rancidity complex No. 1 are respectively radiated, corresponding breeding programs are arranged according to the hereditary characteristics of purple property precursors, full black grain plants are bred from radiation offspring groups, the number of chromosomes is detected through series selection detection and root tip tabletting staining microscopy, and the genetic type of the selected black wheat quality is confirmed. Finally, 4D/4E translocation catamaran line HF-Am3 with 44 chromosomes and 4D/4E translocation replacement line rye line HF-Am4 with 42 chromosomes are bred. The detailed steps of this embodiment are the same as those of embodiment 1, except that the germplasm material is different, and are not described herein again.
Description of the parent materials involved in the above description:
shannon purple wheat No. 1 is examined purple wheat variety in Shandong province; no. 1 of the black horse is a general white-grain wheat variety examined and determined by Shandong province; teacher Luan 02-1 examined the variety of common Baijingjin wheat for the national committee for crop variety approval: loxozhen No. 1 is a purple wheat variety recorded in the late last century of the Committee for examination of variety in Henan province;
blue dwarf septicemia Luan 02-1 and blue dwarf-male-failure black horse No. 1 are both Zibo-ai-people-breed Limited company, the utilizer Luan 02-1, the black horse No. 1 conventional white grain variety is backcrossed with the vernal blue dwarf-male-failure 97-866 (the applicant and the cooperative agreement of the Yi-nationality western-Chang agricultural department of the Sichuan-Liangshan Yao nationality are conditionally obtained) germplasm for continuous backcrossing, and the winterness and semi-winterness blue dwarf-male-failure germplasm (which can be conditionally obtained for compensatory transfer) is gradually bred in the process of genetic background winterization.
Blue dwarf male sterile shannon purple wheat No. 1: zibo lovemin breed Limited utilizes blue dwarf male sterile Luan 02-1 and takes Shannon purple wheat No. 1 as a male parent to carry out continuous backcross transformation to obtain blue dwarf male sterile purple wheat (which can be obtained by conditional transfer).
Blue dwarf-rancidity No. 1: ziboeismin breed limited utilizes blue dwarf male sterile black wheat No. 1 and then uses complex No. 1 as male parent to carry out continuous backcross and transformation to obtain blue dwarf male sterile purple wheat (which can be obtained by conditional paid transfer).
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; those of ordinary skill in the art will understand that: the technical solutions described in the above embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (3)
1. A method for breeding black wheat germplasm with different genetic backgrounds by utilizing blue dwarf male sterile and purple wheat is characterized by comprising the following steps of: backcross pollination is carried out on the blue dwarf-male-sterile wheat germplasm which belongs to a 4D/4E blue grain translocation system monomer by utilizing purple wheat, the blue dwarf-male-sterile purple wheat is transformed into the blue dwarf-male-sterile purple wheat, black grains which are separated from the transformed blue dwarf-male-sterile purple wheat and contain blue grains and purple grain genes and break the linkage are selected, the transformed blue dwarf-male-sterile purple wheat and the selected black grains are subjected to radiation treatment, then the blue dwarf-male-sterile purple wheat is pollinated by the black grains which are subjected to radiation treatment, and black wheat germplasm is bred; the radiation treatment adopts 60 C O -r-rays; the described 60 C O -r-ray dose is 50% to 60% lethal dose; the purple wheat is Shannon purple wheat No. 1 or Loxon No. 1, when the purple wheat is Shannon purple wheat No. 1, the blue dwarf-male-sterile wheat germplasm is blue dwarf-male-sterile Luan 02-1, and when the purple wheat is Loxon No. 1, the blue dwarf-male-sterile wheat germplasm is blue dwarf-male-sterile black horse No. 1; the blue dwarf septicemia Luan 02-1 is a conventional white-grain variety of chef Luan 02-1, is backcrossed with the vernal blue dwarf male parent and is backcrossed with the germplasm of the white dwarf male-sterile 97-866 continuously, and is bred step by step in the winterization process of the genetic background to form the winterness and semi-winterness blue dwarf male-sterile germplasm; the blue dwarf-male-sterile black horse No. 1 is a conventional white-grain variety of the black horse No. 1, is backcrossed continuously to the vernalized blue dwarf-male-sterile 97-866 germplasm, and is gradually bred into the winter-property and semi-winter blue dwarf-male-sterile germplasm in the process of winterization of the genetic background.
2. The method for breeding the black wheat germplasm with the different genetic backgrounds by using the blue dwarf male sterile and the purple wheat according to claim 1, which is characterized in that: the bred black-wheat germplasm comprises 4D/4E translocation double-body black-wheat germplasm and 4D/4E translocation substitution line black-wheat germplasm.
3. The method for breeding rye germplasm with different genetic backgrounds by using blue dwarf male sterile and purple wheat according to claim 2, wherein the method comprises the following steps: the 4D/4E translocation catamaran black wheat germplasm and the 4D/4E translocation replacement line black wheat germplasm both utilize root tip tabletting and observe the chromosome number through microscope microscopic examination to confirm the genetic background type of the bred black wheat.
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