WO2023236392A1

WO2023236392A1 - Method for creating dwarf winter wheat germplasm resources

Info

Publication number: WO2023236392A1
Application number: PCT/CN2022/121799
Authority: WO
Inventors: 刘彦军; 李彩华; 郭家宝; 班进福; 李尹泽
Original assignee: 石家庄市农林科学研究院
Priority date: 2022-06-10
Filing date: 2022-09-27
Publication date: 2023-12-14
Also published as: CN115005100A

Abstract

Disclosed in the present invention is a method for creating dwarf winter wheat germplasm resources. The process comprises: screening high-quality parents for hybridization, performing anther tissue culturing, chromosome doubling and transplantation on an F1 generation, and screening the morphological indicators, cold resistance, physiological indicators and economic indicators of the offsprings according to standards until offsprings complying with the dwarf traits, equivalent to the dwarf winter wheat germplasm resources, are obtained. The method is simple and convenient to operate, can be used for creating a new excellent dwarf winter wheat germplasm, and provides a simple and effective breeding method for wheat breeders.

Description

[Amended in accordance with Rule 26 17.10.2022] A method for creating winter wheat dwarf germplasm resources

Technical field

[Amended in accordance with Rule 26 17.10.2022]
The present invention relates to the technical field of winter wheat dwarf variety screening, and more specifically to a method for creating winter wheat dwarf germplasm resources.

Background technique

Since the world's first "green revolution" initiated by Dr. Borlaug of CIMMYT (International Maize and Wheat Improvement Center) in the 1960s, when the Norin 10 dwarf gene was used for wheat breeding, the research and utilization of dwarf genes have been increasingly used. It is valued by many geneticists and breeding experts. Since Japanese farmers discovered the local dwarf variety "Dharma" in 1935, 5-6 dwarf varieties have appeared in the world. Scientists have conducted genetic identification and analysis of the dwarf genes contained in them. The currently named dwarf gene There are 25, and they are mostly produced by mutations. At present, the dwarf genes in the world's wheat varieties mostly come from Rht1 (Rht-B1b) and Rht2 (Rht-D1b) of "Nonglin No. 10" and Rht8 and Rht9 of red wheat. There are problems with narrow genetic background and single dwarf genes. Therefore, the significance of continuing to discover, identify, and utilize new dwarf sources has been widely valued by countries around the world.

The dwarf parents currently used in wheat breeding have defects such as premature senescence, low grain protein content, small 1,000-grain weight, and low seed setting rate to varying degrees. Therefore, it is particularly important to create and discover new dwarf sources. At the same time, we should also pay attention to the transformation of comprehensive traits of dwarf germplasm resources. If we can innovate a new dwarf source that has the advantages of Norin 10 or Akagomugi dwarf genes and has dominant or partially dominant genetic effects, it will be Get greater utilization in production. With the development of biotechnology, the application of molecular marker technology in wheat dwarf resources will provide more accurate and advanced scientific methods for the creation of wheat dwarf germplasm resources, identification and evaluation of wheat dwarf genes and genetic analysis. The innovative utilization of dwarf germplasm resources has brought about breakthrough progress.

Therefore, how to provide a method for creating winter wheat dwarf germplasm resources is an urgent problem that those skilled in the art need to solve.

Contents of the invention

In view of this, the present invention provides a method for creating winter wheat dwarf germplasm resources. This method combines parent screening and hybrid preparation, hybrid F1 generation anther tissue culture, and determines morphological indicators and physiological indicators of dwarf germplasm resources. and screening criteria are specified. This method is simple and convenient to operate, can be used to create new excellent winter wheat dwarf germplasm, and provides wheat breeders with a simple and effective breeding method.

In order to achieve the above objects, the present invention adopts the following technical solutions:

A method for creating winter wheat dwarf germplasm resources, including the following steps:

Screen high-quality parent crosses, conduct anther tissue culture on the F1 generation, double the chromosomes, and transplant and colonize the offspring. The morphological indicators, cold resistance, physiological indicators, and economic indicators of the offspring are screened according to standards until offspring that meet the dwarf stem characteristics are obtained. That is winter wheat dwarf germplasm resources.

As a preferred technical solution of the above technical solution, the screening indicators for selecting high-quality parents include agronomic traits, disease resistance, cold resistance, economic indicators and quality traits.

Identification and evaluation of agronomic traits: seedling habits, winter and spring characteristics, plant height, panicle length, shell color, awn type, number of grains per panicle, thousand-grain weight, grain color, grain shape, plumpness, heading period, grain filling habit, maturity stage, plant Characteristics such as structure, uniformity, and yellowing properties.

Disease resistance identification and evaluation: Identify its response to diseases under natural disease or inoculation conditions in the field, mainly including resistance to stripe rust, leaf rust, powdery mildew, scab, etc.

Cold resistance identification and evaluation: identify its cold resistance under natural or artificial stress conditions.

Identification and evaluation of economic indicators: identify number of ears per plant, number of grains per panicle, and thousand-grain weight.

Quality trait identification and evaluation: identify the protein, wet gluten, dough rheological properties, and polymer gluten subunit composition of the variety.

The agronomic character screening criteria of the winter wheat parents provided by the screening of the present invention are: plant height ≤ 80 cm, seedlings semi-creeping to creeping, semi-winter to winter, ear length ≥ 7.5 cm, shell color white, ear shape rectangular or spindle-shaped or rod-shaped Ear, number of grains per panicle ≥ 40, thousand-grain weight ≥ 35g, full grains, good gloss, keratin rate greater than or equal to 70%, heading stage and maturity stage ≤ heading stage and maturity stage of national control species, compact or moderate plant type 1-2 , the ear layer is neat, grade 1-2, good yellowing in the later stage, ripeness grade 1-2. It is convenient to breed new germplasm with excellent comprehensive traits.

The identification and evaluation of disease resistance traits of winter wheat germplasm resources provided by the present invention is to identify its response to diseases under natural onset or inoculation conditions in the field, mainly including resistance to stripe rust, leaf rust, powdery mildew, scab, etc. . The disease resistance screening criteria of winter wheat parents provided by the invention are: moderate or above resistance to stripe rust and leaf rust, high resistance or immunity to powdery mildew, moderate or above resistance to scab, and resistance to leaf blight. Convenient to improve disease resistance of newly cultivated varieties

The invention provides identification and evaluation of disease resistance traits of winter wheat germplasm resources, and identifies their cold resistance under natural or artificial stress conditions. The cold resistance screening criteria of winter wheat parent resources provided by the invention: natural overwintering leaf freezing damage is ≤3 levels, and the overwintering dead seedling rate and stem rate in the field are both zero, which facilitates the cultivation of new germplasm with strong cold resistance.

The screening criteria for the economic traits of winter wheat parents provided by the invention are: under single-grain on-demand sowing conditions, the number of ears per plant is ≥ 16 ears, the number of grains per ear is ≥ 40 grains, and the thousand-grain weight is ≥ 35g. It is convenient to ensure the high yield of newly cultivated varieties.

The identification, evaluation and screening criteria for the quality traits of winter wheat parents provided by the invention: the quality of germplasm resources reaches the national standard for strong gluten wheat or the national standard for medium gluten wheat, and its main quality traits: protein is greater than or equal to 13%, wet gluten is ≥30%, medium gluten wheat dough The stabilization time is ≥3 minutes, and the stabilization time of strong gluten wheat dough is ≥7 minutes. It is convenient to ensure the quality improvement of newly cultivated varieties.

According to the KASP marker screening criteria for winter wheat parents provided by the present invention, the selected parents contain one or more pairs of dwarf alleles. It is convenient to ensure the success probability of short-stem germplasm creation.

As a preferred technical solution of the above technical solution, the selected parents are detected by the KASP marker method and contain at least one pair of dwarf alleles.

As the preferred technical solution of the above technical solution, the anther tissue culture of the F1 generation includes:

1) Callus induction: Inoculate the anthers of the hybrid F1 generation spikelets into the callus induction medium, inoculate 25 bottles, inoculate 300-400 anthers in each bottle, and cultivate in the dark at 28-30°C;

2) Culture of differentiated seedlings: When the callus reaches 9-12 days old, transfer it to the differentiated seedling culture medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to obtain seedlings;

3) In-vitro seedling culture: Use the seedling-strengthening medium with paclobutrazol added to cultivate the seedlings for 10-15 days to obtain in-vitro seedlings;

4) Oversummer treatment: When the test tube seedlings grow to 2-3cm, place the test tube seedlings in a low temperature environment of 3-5°C for oversummer cultivation for 90-100 days to obtain tissue culture seedlings.

As the preferred technical solution of the above technical solution, the chromosome doubling and transplanting are as follows: before transplanting, harden the seedlings for 10-15 days. When the temperature drops to 6°C, transplant the tissue culture seedlings to the greenhouse, and then conduct a microscopic examination of the chromosome ploidy , the haploid plants were treated with double colchicine, and the seeds were harvested, which was the H0 generation. The tissue culture seedlings planted in the field have passed the vernalization stage due to the low temperature conditions of refrigeration during the summer. During the winter, they should be protected by small sheds or solar greenhouses to avoid freezing and killing the seedlings, which will affect subsequent selections.

As a preferred technical solution of the above technical solution, the root tips or growth points of haploid plants are treated with 4/10000 colchicine to obtain diploid normal plants.

As the preferred technical solution for the above technical solution, after transplanting and planting, the H0 generation should be managed with water and fertilizer, specifically as follows: after planting, apply 15kg of urea + 20kg of diamine phosphate + 5kg of potassium chloride per mu; topdress 15kg of urea per mu during the jointing stage. /667m ² ; Irrigation should be done five times at base moisture, stand-up, jointing, booting and grouting, with each watering ≥50m ³ .

As the preferred technical solution for the above technical solution, the morphological indicators, cold resistance, physiological indicators and economic indicators are screened according to the standards: plant height ≤ 65cm, plant type is compact or moderate, stems are elastic, and leaves are slender or moderate. , rectangular or spindle-shaped or rod-shaped ears, ≥16 ears per plant, ear layer uniformity level 1-2, number of kernels per ear ≥40, thousand-grain weight ≥35g, full grains, fullness level 1, bright, horniness rate ≥70%, growth period ≤240 days, good yellowing in the later period, maturity level 1-2; frost damage to overwintering leaves ≤3, cold resistance better than or equivalent to the regional trial control species; medium to above resistance to stripe rust and leaf rust , highly resistant or immune to powdery mildew, moderately or above resistant to scab, and resistant to leaf blight; protein ≥ 13%, wet gluten ≥ 30%, dough stabilization time ≥ 3 minutes, KASP marker detection contains 1 or more pairs of dwarfs culm alleles.

Description of the drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings in the following description are only These are embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on the provided drawings without exerting creative efforts.

The attached picture in Figure 1 shows the breeding pedigree of Shi Ai No. 1;

The attached picture in Figure 2 shows the comparison of panicle traits between Shi'ai No. 1 and its parent plants; the left is the parent Yannong 19, the right is the parent Shi 4185, and the middle is Shi'ai No. 1;

Figure 3 is a diagram showing the grain properties of Shi Ai No. 1;

The attached picture in Figure 4 shows the breeding pedigree of Shi Ai No. 2;

Figure 5 is a comparison chart of panicle traits between Shi'ai 2 and its parents; the left is the parent Gao 8901-11-14, the right is the parent Liangxing 99, and the middle is Shi'ai 2;

The attached picture in Figure 6 shows the plant and panicle diagram of Shiai No. 2 during the filling stage;

The attached picture in Figure 7 shows the breeding pedigree of Shi Ai No. 3;

Figure 8 is a diagram showing comparison of panicle traits between Shi'ai No. 3 and its parents. The left is the parent Shijiazhuang No. 8, the right is the parent Shi'ai No. 1, and the middle is Shi'ai No. 3;

Figure 9 is a diagram of the grain properties of Shi Ai No. 3;

The attached picture in Figure 10 shows the field growth chart of Shiai No. 3 population;

The attached picture in Figure 11 shows the breeding pedigree of Shi Ai No. 4;

Figure 12 is a comparison chart of panicle traits between Shiai No. 4 and its parent plants;

Figure 13 is a diagram of Shi Ai No. 4 seeds;

The attached picture in Figure 14 shows the breeding pedigree of Shi Ai No. 5;

The attached picture in Figure 15 is a comparison of panicle traits between Shiai No. 5 and its parents;

Figure 16 is a diagram of Shi Ai No. 5 seeds;

The attached picture in Figure 17 shows the breeding pedigree of Shi Ai No. 6;

The attached picture in Figure 18 shows the comparison of panicle traits between Shiai No. 6 and its parents;

Figure 19 is a breeding pedigree of Shi Ai No. 7;

The attached picture in Figure 20 is a comparison of panicle traits between Shi Ai No. 7 and its parents;

The attached picture in Figure 21 shows the breeding pedigree of Shi Ai No. 8;

The attached picture in Figure 22 is a comparison of panicle traits between Shi Ai No. 8 and its parent plants;

The attached picture in Figure 23 is a grain diagram of Shi Ai No. 8;

The attached picture in Figure 24 shows the breeding pedigree of Shi Ai No. 9;

Figure 25 is a picture of the field appearance of Shidai No. 9 population;

The attached picture in Figure 26 shows the breeding pedigree of Shi Ai No. 10;

Figure 27 is a picture of the field appearance of Shidai No. 10 population;

Figure 28 is a photo of Shi Ai No. 10 seeds.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention.

The test methods used in the following examples are all examples of the invention mentioned above. So far, 10 new dwarf winter wheat germplasm have been successfully created (see Table 1).

Table 1

Example 1 “Shi Ai No. 1” breeding process and breeding method

1) Breeding and characteristics of the female parent “Yan Nong 19”

Yannong 19 is a new wheat variety with high quality, high yield and wide adaptability, which was bred by the Wheat Research Institute of Yantai Academy of Agricultural Sciences using Yan 1933 as the female parent and Shaan 82-29 as the male parent through sexual hybridization and pedigree selection.

Characteristics: winter nature, semi-prostrate seedlings, dark yellow-green leaves, upturned leaves, plant height about 80cm, high tillering rate, medium-large ears, rectangular ears, long awns, white shells, white grains, horny , the number of kernels per ear is about 40, and the thousand-kernel weight is about 40g. It has good yellowing, strong cold resistance and disease resistance, especially high resistance to scab.

2) Breeding and characteristics of the male parent “Shi 4185”:

Shi 4185 is produced by the Shijiazhuang Academy of Agricultural and Forestry Sciences using Taigu core sterile germplasm, aggregating hybridization of Zhi 8094, Baofeng 7228, and Shi 84-7120 and breeding through recurrent selection. From 1997 to 2001, it passed the regional and national variety approvals in Hebei, Henan, Xinjiang and Hotan.

Characteristics: semi-winter, medium ripe. The seedlings are semi-prostrate, with strong tillering ability, high panicle rate, and plant height of about 75 cm. The plant type is compact, the panicle layer is neat, long awns, white shell, rectangular panicle; the panicle length is about 8 cm, the number of grains in the panicle is about 32, the thousand-grain weight is about 38 grams, and the bulk density is more than 790 g/L. Drought-resistant, water-saving, salt-alkali tolerant, stripe rust-resistant, moderately susceptible to leaf rust, lightly susceptible to powdery mildew.

3) The breeding and characteristics of “Shi Ai No. 1”. The breeding method is shown in Figure 1;

Shi Ai No. 1 is a breeding method that combines conventional breeding and anther tissue culture to sexually cross Yannong 19 and Shi 4185. The F1 generation is cultured from anther tissue, and the offspring are bred using directional systematic selection.

The hybrid combination was prepared in May 2003, using "Yan Nong 19" as the female parent and "Shi 4185" as the male parent.

Tissue culture inoculation In mid-April 2004, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 25 bottles were inoculated, with 300-400 anthers in each bottle.

Callus induction was carried out by culturing in the dark at 28-30°C, 350 pieces of callus tissue were induced, and the healing rate was 4.7%.

After the seedlings are differentiated, wait for the callus to reach about 10 days of age. Then transfer it to the differentiation medium for culture. The temperature is 25°C and the light intensity is 1000 Lux. 45 green seedlings are induced. The green seedling differentiation rate is 12.9% and the seedling yield is 0.6%. .

In vitro seedling culture: Use the seedling growth medium with paclobutrazol added to culture the in vitro seedlings to obtain strong seedlings with many roots, many tillers and green leaves. Oversummer treatment: When the test tube seedlings grow to 2-3 cm, place the test tube seedlings in a low temperature environment of 3-5°C for 90-100 days.

Chromosome doubling and transplanting In late November, when the temperature drops to 6°C, the tissue culture seedlings are transplanted to the field greenhouse. Before transplanting, the seedlings were hardened for 10-15 days, and then the chromosome ploidy was examined under a microscope. The haploid plants were double treated with colchicine, and then transplanted and colonized to the protected area of the greenhouse to become the H0 generation.

Field selection of H0 generation: The H0 generation was selected before the summer harvest in 2005. Among them, 5 plants showed short stems, compact plant type, good ear structure, large ears and many grains. When tested after harvest, the plant height is 44-46cm, the ear length is 8-8.5cm, and the number of ears per plant is 8-16. The number of grains per ear is 45-55;

H1 generation selection: In 2005, 5 single plants (clusters) of the selected H0 generation were planted into panicle rows, and a total of 120 rows (H1 generation lines) were planted. In 2006, selection was made based on field performance, among which zp06- Rows 3877-3920 show short stems, large ears, neat ear layers, and good ear structure. The harvested lines will enter the yield ratio test, and the outstanding single plants will be selected to be promoted to H2;

H2 generation selection: In 2007, a single plant was selected to be planted in row zp072-607-730 of the H2 generation. Among them, zp072-689-690 performed outstandingly and was promoted to the yield ratio test at the same time. The yield per mu was 515.6 kg. This strain was named It is "Shidai No. 1".

Characteristics

This germplasm is winter-natured, and its maturity period is equivalent to that of the control "Shi 4185". The plant height of a single seed sown on demand is about 46.6cm (53cm under group conditions). The seedlings are semi-prostrate, with dark green leaves and good cold resistance. The tillering ability is strong and the panicle rate is high, as shown in Figures 2 and 3; the number of panicles per mu in the plot can be as high as 550,000-650,000, and 18-28 panicles per plant can be produced on demand, with neat panicle layers, rectangular panicles, and Good structure, closely arranged spikelets, long awns, white shells, white grains, oval grains, full grains, good gloss, strong firmness, large number of grains per panicle, about 50 grains per panicle (32-35 under population conditions grains), the thousand-grain weight is 32 grams, the grain protein content is 16.1%, and the wet gluten content is 37%. The flag leaves are upturned, the plant shape is compact, the stems are tough and elastic, the root system is developed, and yellowing is good. Short stems and many ears: plant height is about 46.6cm (50-53cm under group conditions); multiple ears: 550,000-650,000 ears per mu; double dwarf stem gene control: according to genetic testing by the Chinese Academy of Sciences, Shi Ai No. 1 contains Rht- B1b Rht-D1b double dwarf gene.

Example 2 “Shi Ai No. 2” breeding process and breeding method

Breeding and characteristics of female parent “Gao 8901-11-14”

Gao 8901-11-14 is a new high-quality wheat variety selected and bred by Gaocheng Agricultural Science and Technology Institute through sexual hybridization of Polish wheat as the female parent and Linzhang wheat as the male parent.

Characteristics: semi-winter, medium-maturing, semi-creeping seedlings, strong tillering ability and high ear-growing rate. The plant height is 80cm, the panicle is spindle-shaped, with short awns, white shells and chaffs with downy hairs, white grains, hard, resistant to stripe rust, leaf rust, and good yellowing. The number of ears per mu is 430,000-450,000, the number of grains per ear is 35, and the weight of 1,000 grains is 38 grams.

Crude protein 15.75%, lysine 0.39%, wet gluten 36.1%, formation time 6.4 minutes, stabilization time 29.2 minutes, bread volume 773ml, bread score 83.3 points.

Breeding and characteristics of the male parent “Liangxing 99”:

Liangxing 99 was bred by Liangxing Research Institute in Dezhou City. The variety comes from "Ji 91102/Lumai 14//PH85-16", which was approved by Hebei Province in 2004.

Characteristics: semi-winter, medium-maturing, semi-creeping seedlings, dark green leaves, strong tillering ability, and high panicle rate. The plant height is about 78 cm, the plant shape is compact, and the flag leaves are raised upward. The ears are spindle-shaped, with long awns, white grains, and horniness. The stems are solid, elastic and resistant to lodging. Mild premature aging, general yellowing. Good cold resistance. Highly resistant to powdery mildew, moderately resistant to slow stripe rust, moderately susceptible to sheath blight, moderately to highly susceptible to leaf rust and stem rust. The number of ears per mu is about 400,000, the number of grains per ear is about 35.7, and the weight of one thousand grains is about 40.0 grams.

The breeding and characteristics of “Shi Ai No. 2” are shown in Figure 4;

Preparation of hybrid combination: In May 2003, sexual cross was carried out with “Gao 8901-11-14” as the female parent and “Liangxing 99” as the male parent.

tissue culture inoculation

In mid-April 2004, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 20 bottles were inoculated, with 300-400 anthers in each bottle.

Callus induction was carried out by culturing in the dark at 28-30°C, and 366 pieces of callus were induced, with a healing rate of 6.1%.

After the seedlings were differentiated, the callus reached 10 days of age and was transferred to the differentiation medium for culture. The temperature was 25°C and the light intensity was 1000 Lux. 76 green seedlings were induced, with a green seedling differentiation rate of 20.7% and a seedling yield of 1.27%.

In vitro seedling culture: Use the seedling growth medium with paclobutrazol added to culture the in vitro seedlings to obtain strong seedlings with many roots, many tillers and green leaves.

Oversummer treatment: When the test tube seedlings grow to 2-3 cm, place the test tube seedlings in a low temperature environment of 3-5°C to overwinter.

Chromosome doubling and transplanting In late November, when the temperature drops to 6°C, the tissue culture seedlings are transplanted to the field greenhouse. Before transplanting, the seedlings are hardened for 10-15 days, and then the chromosome ploidy is examined under a microscope. The haploid plants are treated with double colchicine, and then transplanted and planted in the protected area of the greenhouse to become the H0 generation;

The H0 generation single plants or single ears harvested in the autumn of 2006 were selected and planted in the field as H1 generation lines. The planting row number was ZP07-0091-0295, a total of 205 rows. Among them, ZP07-220-245 showed short stems, multiple ears, and large size. With advantages such as ear, disease resistance, good ear structure, neat ear layer, etc., two excellent lines, ZP07-225 and ZP07-241, were selected to upgrade the next annual yield ratio test. At the same time, the dwarf germplasm was selected as a key line and named. It is called "Shi Ai No. 2". In 2007, outstanding single plants were selected and promoted to the appraisal nursery in 2008 for further appraisal and selection.

The description of the main characteristics of the variety is shown in Figures 5 and 6; semi-winter, mid-late maturing, short stem, the maturity stage is equivalent to the control "Liangxing 99", the plant height is about 54cm, the seedlings are semi-prostrate, dark green leaves, cold resistance Good, strong disease resistance. The tillering ability is strong and the panicle rate is high. According to the plot test, the number of panicles per mu is 450,000-500,000, and the number of panicles per plant is 18-26. The panicle layer is relatively neat, cone-shaped panicle, with downy hulls, long awns, and white Shell, white kernels, oblong kernels, plump kernels, good luster, about 50 kernels per ear (34-38 kernels in a group), and a thousand kernel weight of about 35 grams. The plant type is relatively loose, with tough stems, good elasticity, well-developed root system, and general yellowing. The grain protein content is 15.5%, the wet gluten content is about 35%, and the high molecular gluten subunits are composed of 1, 5+12, 7+8. Double dwarf gene control: According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai 2 contains the Rht-B1b Rht-D1b double dwarf gene.

Example 3 "Shi Ai No. 3" breeding process and breeding method

Breeding and Characteristics of the Female Parent "Shijiazhuang No. 8"

Shijiazhuang No. 8 was developed by the Shijiazhuang Academy of Agricultural Sciences with Shi 91-5065 as the female parent and Shi 9306 (Jimai No. 38) as the male parent. It was bred through alternating directional selection in different places for many years and has the characteristics of water saving, high yield, cold resistance and disease resistance. It is a new winter wheat variety that combines the advantages of good yellowing, good yellowing and so on. .

Characteristics: Semi-winter, medium-maturing, semi-creeping seedlings, strong tillering ability, and more ears per acre. It is cold-resistant, drought-resistant, medium-resistant to stripe rust, highly resistant to leaf rust and powdery mildew, resistant to dry and hot winds, and has a good ripening phase. The plant height is about 75cm, and the stems are tough. Short awns, spindle ears, ear length is about 8cm, number of kernels per ear is about 32, white kernels, hard, weight of 1,000 kernels is about 45 grams, kernels are plump, glossy, and bulk density is 795 grams/liter. .

Breeding and characteristics of the male parent “Shi Ai No. 1”:

Shi Ai No. 1 is a new dwarf wheat germplasm bred by Shijiazhuang Academy of Agricultural and Forestry Sciences.

Characteristics: Winter nature, the maturity stage is equivalent to the control "Shi 4185", the plant height is about 46.6cm (55cm under group conditions), the seedlings are semi-prostrate, the leaves are dark green, and the cold resistance is good. Strong tillering ability, high panicle rate, the number of panicles per mu in the plot can be as high as 500,000-600,000, and 18-28 panicles per plant can be produced on demand. The panicle layer is neat, rectangular, with good panicle structure, closely arranged spikelets, and long Eawn, white shell, white grains, oval grains, full grains, good luster, strong firmness, large number of grains per panicle, about 55 grains (32-35 grains under group conditions), 1,000-grain weight 32-34 grams, The grain protein content is 16.1% and the wet gluten content is 37%. The flag leaves are upturned, the plant shape is compact, the stems are tough and elastic, the root system is developed, and yellowing is good.

The breeding and characteristics of “Shi Ai No. 3” are shown in Figure 7;

The hybrid combination was prepared in May 2008, using "Shijiazhuang No. 8" as the female parent and "Shiai No. 1" as the male parent.

Tissue culture inoculation: In mid-April 2009, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 25 bottles were inoculated, with 300-400 anthers in each bottle.

Callus induction: Cultivated in the dark at 28-30°C, 252 pieces of callus tissue were induced, and the healing rate was 3.36%.

Seedling differentiation: When the callus reaches about 10 days old, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 45 green seedlings. The green seedling differentiation rate is 17.85% and the seedling yield is 0.6 %.

In-vitro seedling cultivation: Use the seedling-strengthening medium added with paclobutrazol to cultivate in-vitro seedlings to obtain strong seedlings with many roots, many tillers and green leaves.

Chromosome doubling and transplanting: In late November 2009, when the temperature dropped to 6°C, the tissue culture seedlings were transplanted to the field greenhouse. Before transplanting, the seedlings are hardened for 10-15 days, and then the chromosome ploidy is examined under a microscope. The haploid plants are treated with double colchicine, and then transplanted and planted in the protected area of the greenhouse to become the H0 generation.

Field selection: Harvested H0 generation single plants or single panicles were planted in the field in the autumn of 2010 to become H1 generation lines. The planting row number was ZP11-0125-0285, a total of 160 rows. Among them, ZP11-125-136 showed short stems and multiple panicles. With the advantages of large panicles, disease resistance, good panicle structure, and neat panicle layers, two excellent lines, ZP11-134 and ZP11-136, were selected to upgrade the yield ratio test next year. At the same time, the dwarf germplasm was selected as a key line and Named "Shi Ai No. 3", outstanding single plants were selected in 2011 and promoted to the appraisal nursery in 2012 for continued appraisal and selection.

For description of the main characteristics of the variety, see Figure 8, Figure 9 and Figure 10; semi-winter, medium-early maturing, short stem, the maturity period is equivalent to the control "Shi 4185", the plant height is about 48cm, the seedlings are semi-prostrate, cold-resistant and disease-resistant , medium tillering power, good plant leaf shape, large ear head, about 400,000 ears per mu, 15-20 ears per plant when sown on demand, tough and elastic stems, well-developed root system, good yellowing property, and neat ear layer. Rectangular ears, long awns, white shells, white grains, full grains, good luster, about 40 grains per panicle (34-38 grains in a group), and a thousand-grain weight of about 42 grams. The grain protein content is 13.5% and the wet gluten content is 30.1%. According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai 3 contains Rht-B1b Rht-D1b double dwarf genes.

Example 4 “Shi Ai No. 4” breeding process and breeding method

Breeding and Characteristics of the Female Parent "Shi 4185" Shi 4185 is a product of the Shijiazhuang Academy of Agricultural and Forestry Sciences using the Taigu nuclear sterile core germplasm to aggregate and cross plant 8094, Baofeng 7228, and Shi 84-7120 and select through recurrent selection. bred. From 1997 to 2001, it passed the regional and national variety approvals in Hebei, Henan, Xinjiang and Hotan.

Characteristics: semi-winter, medium ripe. The seedlings are semi-prostrate, with strong tillering ability, high panicle rate, and plant height of about 75 cm. The plant type is compact, the panicle layer is neat, long awns, white shell, rectangular panicle; the panicle length is about 8 cm, the number of grains in the panicle is about 32, the thousand-grain weight is about 38 grams, and the bulk density is more than 790 g/L.

Breeding and characteristics of the male parent "Shi'ai No. 1": Shi'ai No. 1 is a breeding method that combines conventional breeding and anther tissue culture. Yannong 19 and Shi 4185 are sexually crossed, and the F1 generation is cultured in anther tissue. , the offspring are bred using the method of directional systematic selection.

This germplasm is winter-native, and its maturity stage is equivalent to that of the control "Shi 4185". The plant height is about 46.6cm (55cm under group conditions), the seedlings are semi-prostrate, the leaves are dark green, and they have good cold resistance. It has strong tillering ability and high panicle rate. The number of panicles per mu in the plot can be as high as 500,000-600,000. A single plant can produce 18-28 panicles by sowing on demand. The panicle layer is neat and rectangular. The panicle structure is good and the spikelets are closely arranged and long. Eawn, white shell, white grains, oval grains, full grains, good luster, strong firmness, large number of grains per panicle, about 55 grains (32-35 grains under group conditions), 1,000-grain weight 32-34 grams, The grain protein content is 16.1% and the wet gluten content is 37%. The flag leaves are upturned, the plant shape is compact, the stems are tough and elastic, the root system is developed, and yellowing is good.

The breeding and characteristics of “Shi Ai No. 4”, the breeding map is shown in Figure 11;

Shi Ai No. 4 is a breeding method that combines conventional breeding and anther tissue culture. Shi 4185 and Shi Ai No. 1 are sexually crossed. The F1 generation is cultured in anther tissue, and the offspring are bred using directional systematic selection.

Breeding process and main characteristics of each generation:

Preparation of hybrid combination: In May 2008, a sexual cross was carried out with “Shi 4185” as the female parent and “Shi Ai No. 1” as the male parent.

Tissue culture inoculation: In mid-April 2009, inoculate the anthers of the hybrid F1 generation spikelets onto the callus induction medium, and inoculate 25 bottles with 300-400 anthers in each bottle.

Callus induction: Cultivated in the dark at 28-30°C, 284 pieces of callus tissue were induced, and the healing rate was 3.79%.

Seedling differentiation: When the callus reaches about 10 days old, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 68 green seedlings. The green seedling differentiation rate is 23.9% and the seedling yield is 0.91. %.

Field selection: Harvested H0 generation single plants or single panicles were planted in the field in the autumn of 2010 to become H1 generation lines. The planting row number was ZP11-0300-0406, a total of 106 rows. Among them, ZP11-325-336 showed short stems and multiple panicles. With the advantages of large ears, disease resistance, good ear structure, neat ear layers, etc., three excellent lines, ZP11-325, ZP11-330, and ZP11-328, were selected to upgrade the annual yield ratio test. At the same time, dwarf lines were selected as key lines. The stem germplasm was named "Shi'ai No. 4". In 2011, outstanding individual plants were selected and promoted to the identification nursery in 2012 for continued identification and selection.

For description of the main characteristics of the variety, see Figure 12 and Figure 13; semi-winter, medium-ripening, short stem, mature stage is equivalent to the control "Shi 4185", plant height is about 55cm, seedlings are semi-prostrate, dark green leaves, good cold resistance , strong disease resistance. Strong tillering ability, high panicle rate, 450,000-500,000 panicles per mu, 16-24 panicles per plant when sown on demand, average panicle layer uniformity, rectangular panicles, long awns, white shells, white grains, and uneven grains The seeds are round, plump and glossy. The number of kernels per ear is about 40 (33-35 in a group), and the thousand-grain weight is about 38 grams. The plant shape is moderately tight, the stems are tough and elastic, the root system is developed, and the yellowing is good. The quality is excellent, with grain protein content of 16.2%, wet gluten content of 35.3%, ductility of 167, and sedimentation value of 36.2ml. According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai 4 is controlled by the Rht-B1b and Rht-D1b double dwarf genes.

Example 5 “Shi Ai No. 5” breeding process and breeding method

Breeding and Characteristics of the Female Parent “Yan Nong 19”

Yannong No. 19 is a high-quality strong-gluten wheat variety selected by the Wheat Research Institute of the Yantai Academy of Agricultural Sciences using Yan 1933 as the female parent and Shaan 82-29 as the male parent using the pedigree method. It was approved by the Shandong Provincial Variety Approval Committee in 2001.

Characteristics: This variety is winter-natured, with semi-prostrate seedlings, upturned leaves, and a plant height of about 75cm. It has strong tillering ability, high panicle rate, medium to large panicle, and about 40 grains per panicle. The ears are rectangular, with long awns, white shells, white grains, and horny grains. The grains are round and the thousand-grain weight is about 36 grams. It has strong disease resistance and drought resistance, and medium cold resistance. .

This germplasm is winter-native, and its maturity stage is equivalent to that of the control "Shi 4185". The plant height is about 46.6cm (55cm under group conditions), the seedlings are semi-prostrate, the leaves are dark green, and they have good cold resistance. Strong tillering ability, high panicle rate, the number of panicles per mu in the plot can be as high as 500,000-600,000, and 18-28 panicles per plant can be produced on demand. The panicle layer is neat, rectangular, with good panicle structure, closely arranged spikelets, and long Eawn, white shell, white grains, oval grains, full grains, good luster, strong firmness, large number of grains per panicle, about 55 grains (32-35 grains under group conditions), 1,000-grain weight 32-34 grams, The grain protein content is 16.1% and the wet gluten content is 37%. The flag leaves are upturned, the plant shape is compact, the stems are tough and elastic, the root system is developed, and yellowing is good.

The breeding and characteristics of “Shi Ai No. 5”, the breeding map is shown in Figure 14;

Preparation of hybrid combination: In May 2008, a sexual cross was carried out with “Yan Nong 19” as the female parent and “Shi Ai No. 1” as the male parent.

Tissue culture inoculation: In mid-April 2009, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 10 bottles were inoculated, with approximately 150 anthers in each bottle.

Callus induction: Cultivated in the dark at 28-30°C, 203 pieces of callus tissue were induced, and the healing rate was 13.5%.

Seedling differentiation: When the callus reaches about 10 days old, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 34 green seedlings. The green seedling differentiation rate is 16.75% and the seedling yield is 2.27 %.

Field selection: In the autumn of 2010, the harvested H0 generation single plants or single ears were planted in the field to become H1 generation lines. The planting row number was ZP11 - a total of 160 rows. Among them, ZP11-125-136 showed short stems, multiple ears, and large ears. With the advantages of disease resistance, good panicle structure, neat panicle layer, etc., two excellent lines, ZP11-134 and ZP11-136, were selected to upgrade the next annual yield ratio test. At the same time, the dwarf germplasm was selected as a key line and named " Shidai No. 5".

For description of the main characteristics of the variety, see Figure 15 and Figure 16; semi-winter, medium-early maturing, short stem, maturity period 1-2 days later than the control "Shi 4185", plant height about 54cm (60cm below the population), seedlings half Creeping, cold-resistant, disease-resistant, medium tillering ability, good plant leaf shape, large ear head, 400,000-450,000 ears per mu, 15-20 ears per plant when sown on demand, neat layer of ears, rectangular ears, long It has awns, white shell, and white grains. The grains are full and shiny. The number of grains per ear is about 45 (34-38 grains in a group), and the thousand-grain weight is about 42 grams. The stems are tough and elastic, with well-developed root system and good yellowing resistance. The grain protein content is 14.53%, the wet gluten content is 32.9%, the extensibility is 150, and the sedimentation value is 31.24ml. According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai 5 is controlled by Rht-B1b Rht-D1b double dwarf genes.

Example 6 “Shi Ai No. 6” breeding process and breeding method

Breeding and characteristics of the female parent "Shi'ai No. 1": Shi'ai No. 1 is a new dwarf wheat germplasm bred by the Shijiazhuang Academy of Agricultural and Forestry Sciences.

Breeding and characteristics of "Yan Nong 19": Yan Nong 19 is a high-quality, strong-gluten wheat variety bred by the Wheat Research Institute of the Yantai Academy of Agricultural Sciences using Yan 1933 as the female parent and Shaan 82-29 as the male parent using the pedigree method. , approved by the Shandong Provincial Variety Approval Committee in 2001.

Characteristics: This variety is winter-natured, with semi-prostrate seedlings, upturned leaves, and a plant height of about 81cm. It has strong tillering ability, high panicle rate, medium to large panicle, and about 40 grains per panicle. The ears are rectangular, with long awns, white shells, white grains, and horny grains. The grains are round and the thousand-grain weight is about 36 grams. It has strong disease resistance and drought resistance, and medium cold resistance.

The breeding and characteristics of “Shiai No. 6”, the breeding map is shown in Figure 17;

The hybrid combination was prepared in May 2008, using "Shi'ai No. 1" as the female parent and "Yanyou 19" as the male parent.

Tissue culture inoculation In mid-April 2009, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 1056 anthers were inoculated.

Callus induction: Cultivated in the dark at 28°C, 174 pieces of callus tissue were induced, and the healing rate was 16.48%.

Seedling differentiation: When the callus reaches about 10 days of age, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux. 40 green seedlings will be induced, with a green seedling differentiation rate of 23.00% and a seedling yield of 3.79 %.

Chromosome doubling and transplanting: In late November 2009, when the temperature dropped to 6°C, the tissue culture seedlings were transplanted to the field greenhouse. Before transplanting, the seedlings are hardened for 10-15 days, and then the chromosome ploidy is examined under a microscope. The haploid plants are treated with double colchicine, and then transplanted and planted in the protected area of the greenhouse to become the H0 generation;

Field selection: Harvested H0 generation single plants or single panicles were planted in the field in the autumn of 2010 to become H1 generation lines. The planting row number was ZP11-0546-0679, with a total of 134 rows. Among them, ZP11-0553-0576 showed short stems and multiple panicles. , large panicles, disease resistance, good panicle structure, neat panicle layers, etc., a superior line, ZP11-0555, was selected to upgrade the next annual yield ratio test. At the same time, the dwarf germplasm was selected as a key line and named "Shi." "Ai No. 6", from which outstanding single plants were selected in 2011 and promoted to the identification nursery in 2012 for continued identification and selection.

The main characteristics of the variety are described in Figure 18. It is semi-winter, medium-late maturing, short stem, and the maturity period is 2 days later than the control "Shi 4185". The plant height is about 56cm, the seedlings are semi-prostrate, dark green leaves, and good cold resistance. Strong disease resistance. Strong tillering ability, 400,000-450,000 ears per mu, 15-26 ears per plant after sowing on demand, average ear layer uniformity, rectangular ears, long awns, white shells, white grains, full grains, good gloss, and ears The number of grains is about 50 (35-40 grains in a group), and the weight of one thousand grains is about 42 grams. The plant shape is moderately tight, the stems are tough and elastic, the root system is developed, and yellowing is good. The grain protein content is 13.9%, the wet gluten content is 30%, the extensibility is 149.8, and the sedimentation value is 28.8ml. According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai 6 contains Rht-B1b and Rht-D1b double dwarf genes.

Example 7 “Shi Ai No. 7” breeding process and breeding method

Breeding and Characteristics of the Female Parent "Shimai 15" Shimai 15 was a research project conducted by the Shijiazhuang Academy of Agricultural and Forestry Sciences and the Institute of Plant Physiology of the Hebei Academy of Agricultural Sciences in 1997 using Jimai 38 as the female parent and 92R137 as the male parent. Sexual cross, and rapid propagation breeding technology was used to backcross the F1 generation with Jimai 38 as the male parent, and backcrossed for 4 consecutive generations, and then through field directional selection for 3 consecutive years.

Characteristics: This variety is semi-winter, medium to late maturing. The seedlings are semi-creeping, with medium tillering power, high panicle rate, plant height 70 cm, spindle-shaped panicles, short awns, white shells, white grains, and hard grains. The average number of panicles per mu is 358,000, the number of grains per panicle is 33.6, and the weight of 1,000 grains 42.9 grams. The cold resistance evaluation results in 2009: frost damage level 2, overwintering stems 99.2%, dead stem rate 0.8%. The cold resistance evaluation results in 2010: frost damage level 5, overwintering stems 97.9%, dead stem rate 2.1%. Disease resistance identification results of artificial inoculation by Tianjin Plant Protection Institute: medium resistance to powdery mildew (MR), high susceptibility to leaf rust (HS), and slow stripe rust (SR). Tested by the Ministry of Agriculture's Grain and Products Quality Supervision, Inspection and Testing Center (Harbin): bulk density 784 g/L, crude protein 13.25%, wet gluten 27.3%, sedimentation value 19.5 ml, water absorption 59.4%, formation time 2.0 minutes, stabilization time 1.2 minutes , degree of weakening 234F.U, evaluation value 28, maximum tensile resistance 95E.U, extensibility 12.4 cm, energy 15.3 cm2, hardness index 68.6.

Breeding and characteristics of the male parent "Shan Nong 20": Shan Nong 20 (formerly codenamed Shan Nong 05-066) was a sexual hybrid produced by Shandong Agricultural University with PH82-2-2 as the female parent and 954072 as the male parent. , a new multi-resistant and high-yield wheat variety bred by pedigree method combined with molecular marker-assisted selection. From 2006 to 2007, it participated in the preliminary trials of Huanghuainan tablets, and from 2007 to 2009, it participated in the national regional trials and production trials. In 2010, it completed the trial procedures and passed the national approval (approval number: National Approval Mai 2010006).

Characteristics: A mid-winter, mid-late maturing variety, with an average maturity period of about 1 day later than the control stone 4185. The seedlings are creeping and have strong tillering ability. Regional field trials have documented better overwintering cold resistance. Spring development is steady, polarization is fast, heading is slightly late, there are many ears per acre, and the ear layer is neat. The plant height is 78 cm, the plant shape is compact, the flag leaf is raised upward, and the leaf color is dark green. Good lodging resistance. Later maturation and yellowing are normal. The ears are spindle-shaped, with long awns, white shells, and white grains, which are horny and plump. The number of ears per mu is 433,000 ears, the number of grains per ear is 35.1, and the weight of 1,000 grains is 41.4 grams. Cold resistance identification: Cold resistance is poor. Disease resistance identification: Highly susceptible to scab, sheath blight, moderately susceptible to powdery mildew, slow stripe rust, moderately resistant to leaf rust. The quality measurement results in 2009 and 2010 were respectively: grain bulk density 828 g/L, 808 g/L, hardness index 67.7 (2009), protein content 13.53%, 13.3%; flour wet gluten content 30.3%, 29.7%, sedimentation Value 30.3 ml, 28 ml, water absorption 64.1%, 59.8%, stabilization time 3.2 minutes, 2.9 minutes, maximum tensile resistance 256E.U, 266E.U, extensibility 133 mm, 148 mm, tensile area 47 square centimeters, 56 square centimeters.

The selection process and the main characteristics of each generation are shown in 191 for the selection map;

The hybrid combination was prepared in May 2010, using "Shimai 15" as the female parent and "Shannong 20" as the male parent.

Tissue culture inoculation: In mid-April 2011, inoculate the anthers of the hybrid F1 generation spikelets onto the callus induction medium, inoculate 25 bottles, and inoculate 300-400 anthers in each bottle.

Callus induction: Cultivate in the dark at 28-30°C, 340 pieces of callus tissue were induced, and the healing rate was 4.5%.

Seedling differentiation: When the callus reaches about 10 days old, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 45 green seedlings. The green seedling differentiation rate is 12.9% and the seedling yield is 0.6 %.

Chromosome doubling and transplanting: In late November, when the temperature drops to 6°C, the tissue culture seedlings are transplanted to the field greenhouse. Before transplanting, the seedlings were hardened for 7 days, and then the chromosome ploidy was examined under a microscope. The haploid plants were treated with double colchicine, and then transplanted and colonized to the protected area of the greenhouse to become the H0 generation.

Field selection: H0 generation selection: The H0 generation was selected before the summer harvest in 2012. Among them, 6 plants showed short stems, compact plant type, good disease resistance, and good panicle structure. After harvest, the plant height was 45-49cm, the ear length was 7.5-8cm, the number of ears per plant was 8-16, and the number of grains per ear was 40-52.

H1 generation selection: In 2012, 5 single plants (clusters) of the selected H0 generation were planted into ear rows, and a total of 120 rows (H1 generation lines) were planted. In 2013, selection was based on field performance, among which the H13- Rows 001-020 have short stems, neat panicle layers, good panicle structure, good disease resistance (immune to powdery mildew), and good yellowing. Harvest strain H13-017 and enter the yield ratio test, and select outstanding individual plants to be promoted to H2.

H2 generation selection: In 2014, selected single plants will be planted in the H14-207-230 row of the H2 generation. Among them, the H13-017 selected strain H14-207-212 has outstanding performance. At the same time, the H13-017 selected for yield ratio test identification has the following performance: Under group conditions, the comprehensive tillering ability is strong, the panicle rate is high, disease resistance is good, the panicle layer is neat, and the comprehensive traits are good. The yield per mu is 525.8 kg, which is the same as the adjacent control. At the same time, the research team evaluated its water-saving and drought resistance. The results showed that this strain has a developed root system, outstanding drought resistance, and has a certain tolerance to both soil and atmospheric drought. The strain was named "Shi'ai No. 7"

The description of the main characteristics of the variety is shown in Figure 20; semi-winter, mid-late maturing, short stem, maturity period is 1 day later than the control "Shi 4185", plant height is about 48cm, seedlings are creeping, leaves are green, good cold resistance, disease resistance Stronger (the natural identification of powdery mildew in the field is mild, and it is a highly resistant type). Strong tillering ability, high panicle rate, 450,000-500,000 panicles per mu, 16-24 panicles per plant when sown on demand, relatively neat panicle layer, spindle-shaped panicles, long awns, white shells, white grains, good grain luster, and fine panicle The number of grains is about 38 (30-33 grains in a group), and the weight of one thousand grains is about 45 grams. The plant type is compact, the stems are tough and elastic, the root system is developed, and the yellowing resistance is good. The grain protein content is 14.54%, the wet gluten content is 31.7%, the extensibility is 145.8, and the sedimentation value is 33ml. According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai 7 contains Rht-B1b Rht-D1b double dwarf genes.

Example 8 “Shi Ai No. 8” breeding process and breeding method

Breeding and characteristics of female parent “Shi H083-366”

Breeding and characteristics of the male parent “Shi Ai No. 1”:

The breeding and characteristics of "Shi'ai No. 8" are shown in Figure 21;

Preparation of hybrid combination: In May 2010, sexual hybridization was carried out with “H083-366” as the female parent and “Shi Ai No. 1” as the male parent.

Tissue culture inoculation: In mid-April 2011, inoculate the anthers of the hybrid F1 generation spikelets onto the callus induction medium, inoculate 30 bottles, and inoculate 300-400 anthers in each bottle.

Callus induction: Cultivate in the dark at 28-30°C, 155 pieces of callus tissue were induced, and the healing rate was 1.7%.

Seedling differentiation: When the callus reaches about 10 days old, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 32 green seedlings. The green seedling differentiation rate is 20.6% and the seedling yield is 0.36 %.

Chromosome doubling and transplanting: In late November 2011, when the temperature dropped to 6°C, the tissue culture seedlings were transplanted to the field greenhouse. Before transplanting, the seedlings were hardened for 7 days, and then the chromosome ploidy was examined under a microscope. The haploid plants were treated with double colchicine, and then transplanted and colonized to the protected area of the greenhouse to become the H0 generation.

Field selection: In the autumn of 2012, the harvested H0 generation single plants or single ears were planted in the field to become H1 generation lines. The planting row number was H13-0195-0255, a total of 60 rows. Among them, H13-0240-0255 showed short stems and multiple ears. With the advantages of large ears, disease resistance, good ear structure, and neat ear layers, two excellent lines, H13--245 and H13-246, were selected to upgrade the yield ratio test next year. At the same time, they were selected as key lines for dwarf germplasm. And named it "Shidai No. 8".

The description of the main characteristics of the variety is shown in Figure 22 and Figure 23. It is semi-winter, mid-late maturing, and short-stem. The maturity period is 1-2 days later than the control "Shi 4185". The plant height is about 45cm. The seedlings are semi-prostrate, with green leaves and cold resistance. It has good sex and strong disease resistance (naturally identified as immune to powdery mildew in the field). Strong tillering ability, high panicle rate, about 450,000 panicles per mu, 16-25 panicles per plant when sown on demand, neat panicle layer, rectangular panicle, long awn, white shell, white grains, good grain luster. The number of grains per ear is about 40 (33-35 grains in the case of a group), and the thousand-grain weight is about 48 grams. The plant type is compact, the stems are tough and elastic, the root system is developed, and the yellowing property is average. The grain protein content is 14.36%, the wet gluten content is 32.52%, the extensibility is 142.3, and the sedimentation value is 31ml. According to the KASP gene marker test of the Chinese Academy of Sciences, Shi Ai No. 8 contains Rht-B1b Rht-D1b double dwarf genes.

Example 9 “Shi Ai No. 9” breeding process and breeding method

The breeding and characteristics of the male parent “Shi Ai No. 1”;

The breeding and characteristics of “Shi Ai No. 9” and the breeding method are shown in Figure 24

Preparation of hybrid combination: In May 2013, sexual hybridization was carried out with “Heng 2011-5007” as the female parent and “Shiai No. 1” as the male parent.

Tissue culture inoculation: In mid-April 2014, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 12 bottles were inoculated, with 300-400 anthers in each bottle.

Callus induction: Cultivate in the dark at 28-30°C, induce 90 pieces of callus tissue, and the healing rate is 2.5%.

Seedling differentiation: When the callus reaches about 10 days of age, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 30 green seedlings, with a green seedling differentiation rate of 33% and a seedling yield of 0.83 %.

Chromosome doubling and transplanting: In late November, when the temperature drops to 6°C, the tissue culture seedlings are transplanted to the field greenhouse. Before transplanting, the seedlings were hardened for 7 days, and then the chromosome ploidy was examined under a microscope. The haploid plants were treated with double colchicine, and then transplanted and planted into the protected area of the greenhouse to become the H0 generation.

Field selection of H0 generation: The H0 generation was selected before the summer harvest in 2015. Among them, 10 plants showed short stems and ears, good plant structure, good disease resistance, large ears, and good ear structure. When tested after harvest, the plant height is 47-50cm, the ear length is 8.5-9cm, and the number of ears per plant is 12-16. Number of grains per ear: 45-50 grains

H1 generation selection: In 2016, 10 single plants (clusters) of the selected H0 generation were planted into ear rows, and a total of 80 rows (H1 generation lines) were planted. In 2016, selection was based on field performance, among which the H16- The two rows 1099-3 and H16-1100-3 have short stems, neat panicle layers, good panicle structure, good disease resistance, and good yellowing. The selected harvest lines will enter the yield ratio test, and the best plants will be selected to be promoted to H2 for continued identification.

In 2017, the strain H16-1099-3, which was promoted to the yield ratio test, performed outstandingly, with a yield of 535.8kg per mu, which was the same as the adjacent control. It also showed short stems and large ears, disease resistance and high yield, and neat panicle layers. This strain was named For "Shidai No. 9"

The description of the main characteristics of the variety is shown in Figure 25; it is semi-winter, medium-ripening, short-stem, and the maturity stage is equivalent to the control "Shi 4185". The plant height is about 50cm, the seedlings are semi-prostrate, green leaves, good cold resistance, and relatively disease resistance. Strong (the natural identification of powdery mildew in the field is mild, and it is a highly resistant type). Strong tillering ability, 400,000-450,000 ears per mu, 15-22 ears per plant after sowing on demand, relatively neat panicle layer, spindle-shaped ears, long awns, white shells, white grains, good gloss, and 45-22 grains per panicle. 50 pieces (35-40 grains in the case of a group), and the weight of 1,000 grains is about 45 grams. The plant type is compact, the stems are tough and elastic, the root system is developed, and the yellowing resistance is good. The grain protein content is 15.1%, the wet gluten is 32.3%, the extensibility is 153.2, and the sedimentation value is 28.6ml.

Example 10 Breeding of “Shi Ai No. 10”

Breeding and characteristics of the female parent "Shi'ai No. 6": semi-winter, mid-late maturing, short stem, and comparison of maturity stages

Photo "Shi 4185" was taken 2 days later, the plant height is about 56cm, the seedlings are semi-prostrate, the leaves are dark green, and they are cold resistant.

Good, strong disease resistance. Strong tillering ability, 400,000-450,000 ears per mu, 15-26 ears per plant when sown on demand,

Layer uniformity is average, rectangular ears, long awns, white shells, white grains, full grains, good luster, ears

The number is about 50 (35-40 grains in a group), and the weight of one thousand grains is about 42 grams. The plant type is moderately tight,

The stems are tough and elastic, with well-developed root system and good yellowing. Kernel protein content 13.9%, wet gluten content

The weight is 30%, the ductility is 149.8, and the sedimentation value is 28.8ml.

Breeding and characteristics of the male parent "H083-366": "H083-366" was produced by the Shijiazhuang Academy of Agricultural Sciences using Shi 03-4391 as the female parent and "Shi B03-5455" as the male parent. The F1 generation was cultured from anther tissue. New winter wheat germplasm with high yield, disease resistance and strong stems obtained through backward selection.

Characteristics: Semi-winter, medium-maturing, semi-creeping seedlings, strong tillering ability, and more ears per acre. The plant height is about 78cm, the stem has good toughness and is strong and resistant to lodging. The plant has good leaf shape, good yellowing, and good yield. Disease resistance: immune to powdery mildew, resistant to stripe rust, leaf rust and leaf blight; insect resistance: mild aphids, resistant to midges.

(3) The breeding and characteristics of “Shi Ai No. 10”. The breeding method is shown in Figure 26;

Preparation of hybrid combination: In May 2011, a sexual cross was carried out with "Shi'ai No. 6" as the female parent and "H083-366" as the male parent.

Tissue culture inoculation: In mid-April 2012, the anthers of the hybrid F1 generation spikelets were inoculated onto the callus induction medium, and 24 bottles were inoculated, with 300-400 anthers in each bottle.

Callus induction: Cultivate in the dark at 28-30°C, induce 300 pieces of callus tissue, and the healing rate is 4.2%.

Seedling differentiation: When the callus reaches about 10 days old, transfer it to the differentiation medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to induce 23 green seedlings. The green seedling differentiation rate is 11.5% and the seedling yield is 0.32 %.

Field selection: H0 generation selection: The H0 generation was selected before the summer harvest in 2013. Among them, 6 plants showed short stems, compact plant type, good disease resistance, and good panicle structure. After harvest, the plant height was 46-48cm, the ear length was 8-8.5cm, the number of ears per plant was 8-18, and the number of grains per ear was 43-45.

H1 generation selection: In 2014, 5 single plants (clusters) of the selected H0 generation were planted into ear rows, and a total of 120 rows (H1 generation lines) were planted. In 2014, selection was based on field performance, among which the H14- Rows 025-040 have short stems, neat panicle layers, good panicle structure, and good yellowing. Select outstanding single plants and advance them to H2.

H2 generation selection: In 2015, single plants will be selected to be planted in the H15-107-160 row of the H2 generation. Among them, H15-158 and H15-207 have outstanding performance, with strong comprehensive tillering ability, high panicle rate, good disease resistance, and panicle layer. Neat, with good overall properties.

H3 generation selection: In 2016, single plants will be selected to be planted in rows H16-3001-3008 of the H3 generation. Among them, H16-3005 has outstanding performance, with strong comprehensive tillering ability, high panicle rate, good disease resistance, neat panicle layer, and comprehensive traits. good. The yield ratio test was carried out. After testing, the yield of H16-3005 per mu was 533.6 kilograms, which was 3.85% higher than the adjacent control. It also showed good comprehensive traits, good yield, short stems and large grains, with a thousand-grain weight of 54g. It has outstanding advantages as a seed resource. Selected as short stem and large grain resource, the release number is "H16-3005" and named "Shi Ai No. 10"

The description of the main characteristics of the variety is shown in Figure 27 and Figure 28; it is semi-winter, medium-ripening, and short-stemmed. The maturity stage is equivalent to the control "Shi 4185". The plant height is about 47cm. The seedlings are semi-prostrate, with green leaves, good cold resistance, and resistance to cold. The disease is relatively strong (the natural identification of powdery mildew in the field is mild, and it is a highly resistant type). Strong tillering ability, high panicle rate, 500,000-600,000 panicles per mu, 16-30 panicles per plant when sown on demand, relatively neat panicle layer, spindle-shaped panicles, long awns, white shells, white grains, good grain luster, and fine panicle The number of grains is about 40 (30-33 grains in a group), and the weight of one thousand grains is about 54 grams. The plant type is compact, the stems are tough and elastic, the root system is developed, and the yellowing resistance is good. The grain protein content is 13.76%, the wet gluten content is 29.8%, the extensibility is 149.1, and the sedimentation value is 28.4ml.

Each embodiment in this specification is described in a progressive manner. Each embodiment focuses on its differences from other embodiments. The same and similar parts between the various embodiments can be referred to each other.

The above description of the disclosed embodiments enables those skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be practiced in other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

A method for creating winter wheat dwarf germplasm resources, which is characterized by including the following steps:

Screen high-quality parent crosses, conduct anther tissue culture on the F1 generation, double the chromosomes, and transplant and colonize the offspring. The morphological indicators, cold resistance, physiological indicators, and economic indicators of the offspring are screened according to standards until offspring that meet the dwarf stem characteristics are obtained. That is winter wheat dwarf germplasm resources.
A method for creating winter wheat dwarf germplasm resources according to claim 1, characterized in that the screening indicators for screening high-quality parents include agronomic traits, disease resistance, cold resistance, economic indicators and quality traits.
A method for creating winter wheat dwarf germplasm resources according to claim 1, characterized in that the selected parents are detected by the KASP marker method and contain at least one pair of dwarf alleles.
A method for creating winter wheat dwarf germplasm resources according to claim 1, characterized in that performing anther tissue culture on the F1 generation includes:

1) Callus induction: Inoculate the anthers of the hybrid F1 generation spikelets into the callus induction medium, inoculate 25 bottles, inoculate 300-400 anthers in each bottle, and induce callus tissue by dark culture at 28-30°C.

2) Culture of differentiated seedlings: When the callus reaches 9-12 days old, transfer it to the differentiated seedling culture medium and culture it at a temperature of 25°C and a light intensity of 1000 Lux to obtain seedlings and induce green seedlings.

3) In-vitro seedling cultivation: Use seedling-strengthening medium with paclobutrazol added to cultivate the seedlings for 10-15 days to obtain in-vitro seedlings.

4) Oversummer treatment: When the test tube seedlings grow to 2-3cm, place the test tube seedlings in a low temperature environment of 3-5°C for oversummer cultivation for 90-100 days to obtain tissue culture seedlings.
A method for creating winter wheat dwarf germplasm resources according to claim 4, characterized in that doubling the chromosomes and transplanting and colonizing are: before transplanting, harden the seedlings for 10-15 days, and perform tissue culture when the temperature drops to 6°C. The seedlings were transplanted and colonized in the greenhouse, and then the chromosome ploidy was examined under a microscope. The haploid plants were double treated with colchicine, and the seeds were harvested to form the H0 generation.
A method for creating winter wheat dwarf germplasm resources according to claim 5, characterized in that, haploid plant root tips or growing points are treated with 4/10000 colchicine for 8 hours to obtain diploid normal plants. .
A method for creating winter wheat dwarf germplasm resources according to claim 6, characterized in that water and fertilizer management is performed on the H0 generation, specifically: after planting, apply 15kg of urea + 20kg of diamine phosphate + 5kg of potassium chloride per mu; Apply 15kg/667m 2 of urea per acre from the starting stage to the jointing stage; irrigation should be done five times including plantar moisture, starting, jointing, booting and grouting, with each watering ≥50m 3 .
A method for creating winter wheat dwarf germplasm resources according to claim 7, characterized in that screening morphological indicators, cold resistance, physiological indicators and economic indicators according to standards is specifically: H0 generation, H1 generation, H2 generation plant height is ≤65cm, plant type is compact or moderate, stems are elastic, leaves are slender or moderate, rectangular or spindle-shaped or rod-shaped ears, single plant has ≥16 ears, and ear layer uniformity is 1-2. The number of kernels per ear is ≥40, the thousand-kernel weight is ≥35g, the kernels are plump, plumpness level 1, bright, cutin rate ≥70%, growth period ≤240d, good yellowing in the later period, ripeness level 1-2; overwintering leaf frost damage ≤level 3 , cold resistance is better than or equivalent to the regional trial control species; medium to above resistance to stripe rust and leaf rust, high resistance or immunity to powdery mildew, above medium resistance to scab, resistance to leaf blight; protein ≥ 13%, Wet gluten ≥30%, dough stabilization time ≥3 minutes, KASP marker detection contains 1 or more pairs of dwarf alleles.