CN106613902B - Method for breeding early-maturing new variety by using CMS and DH lines of cabbage - Google Patents

Abstract

The invention discloses a method for breeding a new early-maturing variety by utilizing cabbage CMS and DH lines, which comprises the following steps of selecting a cabbage cytoplasmic male sterile line DH08-GW100M3CMS as a female parent and selecting a DH line DH12-3-M2 as a male parent to carry out hybridization to obtain a DH08-GW100CMS multiplied by DH12-3-M2 combination, and naming the combination as Qinlian 1658 variety. The variety is subjected to variety comparison tests, variety region tests and production demonstration in main vegetable cultivation regions such as Yangling, Xian and Baoji Taibai in Shaanxi province, the heterosis is remarkable, the comprehensive economic character is excellent, the bulb cracking resistance is high, the high-quality disease resistance and the high yield are outstanding, the variety adaptability is strong, the yield increase advantage is stable, and the yield increase amplitude is 10.2% -14.3% compared with that of a control cabbage variety 8398.

Description

Method for breeding early-maturing new variety by using CMS and DH lines of cabbage

Technical Field

The invention belongs to the discipline of crop genetic breeding in agricultural science and technology, and particularly relates to a method for breeding a new early-maturing variety by utilizing a cabbage CMS and a cabbage DH line.

Background

The cabbage is one of the main vegetable varieties in China, contains rich nutrient substances and anticancer substances, and plays an important role in promoting the health of human beings and improving the living standard of people. Spring cabbage occupies almost half of the area in cabbage cultivation in China, and is the main flower color type of spring vegetable supply; the spring cabbage cultivation variety is mainly selected from early-maturing varieties for cultivation. The crack-prone sphericity of early-maturing varieties of spring cabbages seriously hinders the development of the cabbage industry, the fresh vegetable market supply period and the inconsistency of head joints among cabbage plants, so that the harvest asynchronism of the same-field cabbages is easily caused, and the planting benefit of unit area is influenced; the analysis is mainly caused by the lack of breeding resources of the resistant blastomere and the high gene non-homozygosity of the inbred line of the traditional artificial multi-generation inbred breeding. According to the industrial requirements of the cabbages, the spring cabbages are orderly arranged to be resistant to cracking and are listed as the breeding target of new varieties by the national twelve and five; the uniformity of the cabbage excellent hybrid is a prerequisite for production, popularization and application, and is also a standard for measuring the high quality of the hybrid. Therefore, according to the breeding target of the national spring cabbage, a haploid (DH) line with excellent target characters is created and selected by utilizing a cabbage microspore culture technology and an excellent DH line field identification target, and a new spring cabbage variety with crack resistance and highly ordered characters is prepared and bred in a combined mode through a heterosis utilization way, so that the research task target of the people is achieved.

Disclosure of Invention

Aiming at the problems and defects in the prior art, the invention aims to provide a method for breeding a new early-maturing cabbage variety by using a cabbage dihaploid DH line, and the new cabbage variety Qinlian 1658 bred by the method is obvious in heterosis, excellent in comprehensive economic character, excellent in resistance to cracking, and disease resistance and stable in yield-increasing advantage through variety comparison tests, variety region tests and production demonstration.

In order to achieve the purpose, the technical scheme adopted by the invention is a method for preparing a new anti-cracking-ball early-maturing variety by using a cabbage Dihaploid (DH) line, which specifically comprises the following steps:

1) breeding of female parent germplasm material DH08-GW100M3CMS

The female parent DH08-GW100M3CMS cytoplasmic male sterile line is formed by hybridizing an Ogura cytoplasmic cabbage male sterile line serving as a female parent material with a cabbage excellent DH08-GW100M3 line and then carrying out 5-generation backcross transformation;

the female parent maintaining line DH08-GW100M3 is a DH line which is a perfect double haploid line of a crack-resistant bulb and is created by culturing microspores of cabbage early-maturing hybrids 'hope' by a cabbage isolated microspore culture technology.

2) Breeding of male parent germplasm material DH12-3-M2

The male parent DH12-3-M2 is disease-resistant high-quality DH12-3-M2 obtained by culturing early-maturing cabbage variety Qinlian 50 with free microspore to obtain DH12-3 strain and selfing.

3) Selection of hybrid seeds

A cabbage cytoplasmic male sterile line DH08-GW100M3CMS is selected as a female parent and a DH line DH12-3-M2 is selected as a male parent to be hybridized and bred into a DH08-GW100M3CMS xDH 12-3-M2 combination which is named as Qinlian 1658 variety.

Through variety comparison test, variety region test and production demonstration, the Qinlian 1658 is subjected to variety comparison test and demonstration in the main vegetable cultivation regions of Shaanxi province, such as Yangling, Xian and Baoji Taibai, and the like, has the advantages of remarkable heterosis, excellent comprehensive economic character, outstanding crack resistance, high quality and disease resistance, stable yield increase advantage and 10.2-14.3% of yield increase compared with the control cabbage 8398 variety.

The Qinlian 1658 is highly resistant to virus and black rot, resistant to cracking of the bulbs and good in leaf bulb commodity, and the appearance of the leaf bulbs accords with the habit of consumers; the ball leaf slices are crisp and tender in texture, and are fragrant and sweet in raw food.

Drawings

FIG. 1 is a diagram of female parent cabbage cytoplasmic male sterile line DH08-GW100M3CMS breeding.

FIG. 2 is the characteristic diagram of female parent cabbage cytoplasmic male sterile line DH08-GW100M3 CMS.

FIG. 3 is the breeding diagram of male parent DH line DH 12-3-M2.

FIG. 4 shows the characteristics of DH12-3-M2 of male parent.

FIG. 5 is the breeding diagram of the first filial generation Qinlan 1658 variety of cabbage.

FIG. 6 is the character map of the first filial generation Qinlan 1658 variety of cabbage.

Detailed Description

The advantageous effects of the present invention will be described in further detail below by specific examples and specific area tests given by the inventors.

Example 1

1. Sources of materials

1.1 Breeding of female parent germplasm material DH08-GW100M3CMS

The female parent DH08-GW100M3CMS cytoplasmic male sterile line is formed by hybridizing a cabbage Ogura cytoplasmic male sterile line and a maintainer line DH08-GW100 in 2008 and carrying out 5-generation backcross transformation; the female parent line has stable characters, thorough sterility, normal flowering, developed honey glands, degenerated stamens, normal pistils, strong plant growth, no yellowing at low temperature and no pollen grains, and all plants and flower organs are normal at normal temperature except sterility; precocious maturity, planting until harvesting for 58 days; the leaf ball is round, the central column is 5.6cm, the compactness is 0.65, the outer leaf is yellow green, the wax powder is medium, the disease and virus resistance and black rot resistance are realized in open field cultivation in spring, the dry burning core is resisted, the ball cracking resistance is high, the winter property is high, and the combining force is high. The branching of the seed plants is strong, the average height of the seed plants is 86.3cm, and the flowering period is 38-40 days.

The female parent maintaining line DH08-GW100 is a DH line created by culturing microspores of cabbage early-maturing hybrids through a cabbage isolated microspore culture technology.

A breeding technical scheme (figure 1) of a female parent germplasm material DH08-GW100M3CMS and the performance of the characters thereof (figure 2).

1.2 Breeding of male parent germplasm material DH12-3-M2

The male parent DH12-3-M2 is created by utilizing free microspores of a cabbage early-maturing variety Qinlian 50, a DH12-3 strain is obtained in 2010, and a DH12-3-M2 line is obtained in 2011. Through the identification of the agronomic characters in the field in 2011-2012, the characters bred in 2012 are highly regular, the genetic inheritance is stable, and the haploid inbred line has strong disease resistance; the maturity is early, the leaves are yellow green and less in wax powder after being planted for 52 days; the leaf ball is round, the central column is 4.6cm, and the compactness of the leaf ball is 0.64; resisting virus, black rot, dry burning, and cracking; in the reproductive growth period, the plant branching is strong, the average plant height is 100.8cm, the flowering period is about 35 days, the flower organ is normal, the pollen amount is large, the combining force is high, a breeding map of a male parent DH line DH12-3-M2 is shown in figure 3, and a character map of a male parent DH line DH12-3-M2 is shown in figure 4.

1.3 selection of hybrid seeds

Is bred by hybridizing a cabbage cytoplasmic male sterile line DH08-GW100M3CMS and a DH line DH12-3-M2, and is named as Qinlian 1658.

1.3.1 characteristic features of Qinlian 1658:

qinlian 1658(DH08-GW100CMS X DH12-3-M2) is a first generation variety of early-maturing hybrids of spring cabbage, and is planted until harvested for about 58 days; the plant type is semi-vertical, the outer leaves are green, the leaf surfaces are smooth, the wax powder is less, and the leaf balls are emerald green; the plant expansion degree is 38.2-39.1 cm, the number of outer leaves is 10-11, the weight of a single sphere is about 1.2kg, the leaf sphere is circular, the compactness of the leaf sphere is 0.70, the length of a central column is 5.1cm, the height of the central column is less than half of the sphere, and the leaf-leaf ratio is 20.9%. High virus resistance, black rot, dry heartburn, bulb cracking resistance and strong winter property, wherein a cabbage hybrid generation Qinlian 1658 variety breeding diagram is shown in a figure 5, and a cabbage hybrid generation Qinlian 1658 variety character diagram is shown in a figure 6.

1.3.2 variety quality and resistance to blastomere of Qinlian 1658:

the leaf ball has good commodity, and the appearance of the leaf ball accords with the habit of consumers; the ball leaf slices have crisp and tender texture, fragrant and sweet raw food and excellent flavor and quality.

The analysis of the main nutrient components: 1.95% of crude fiber, Vc 50.99mg/100g, 2.55% of soluble sugar and 5.70% of dry matter.

The physiological maturity and late harvest of the phyllosphere 5 celestial sphere index is less than grade 3, the ball cracking rate is less than 5 percent, and the ball cracking resistance reaches high grade.

1.3.3 suitable regions and yield levels

The Qinlan 1658 variety is a spring cabbage cracking-resistant early-maturing variety, has early maturity and strong stress resistance, is suitable for spring cultivation in northern areas of China, and has the yield level of 3739.3kg/667m²。

1.3.4 technical points of cultivation

1) Sowing and seedling raising, namely, in northwest areas, in open field in spring, sowing and seedling raising can be carried out in positive ridges in the middle and last ten days of 12 months or in greenhouses in the middle and last ten days of 1 month, and soil lumps are planted when 6-7 true leaves are planted.

2) Planting: and (3) performing open field cultivation, namely after the land is completely thawed in early spring, keeping the temperature to be more than 10 ℃, and planting seedlings when 6-7 true leaves exist. The air temperature is lower when seedlings are planted in spring, and the seedlings are suitable for planting in water by using the seedling-planting seat, which is beneficial to improving the ground temperature. Flat bed or half-high bed cultivation is adopted, and the density is 4000-4500 seedlings per square meter 667; otherwise, the cultivation is too dense or too sparse, which results in reduced yield or untight bulbs.

3) Field management:

filling water: irrigating once after field planting and seedling revival, wherein the air temperature and the ground temperature are relatively low, and the irrigation quantity is not suitable to be too large. Then controlling irrigation or irrigating water once for a while due to over-dry soil until the squat seedlings reach the initial stage of balling. After the core is wrapped, the temperature is raised, the irrigation quantity needs to be increased, and the irrigation frequency is increased; when the ground is dry, the water should be poured. The water is irrigated for 4-5 times in the growing period, and the water is properly controlled before harvesting to prevent the balls from cracking and influence the yield and the product quality.

Secondly, fertilizing: and (4) topdressing for 3-4 times. The nitrogen, phosphorus and potassium are used as main materials in the early stage of top dressing, and the nitrogen fertilizer is used as a main material in the later stage. Applying base fertilizer again, wherein 5000kg of organic fertilizer and 50kg of compound fertilizer are applied per mu; after planting, intertillage and topdressing are carried out once after about 10 days, 5 kilograms of urea is topdressed per mu, and sufficient nutrients for growth of the rosette leaves are provided. In the initial stage of lotus seedpod application, top dressing is performed in holes, the fertilizing concentration is improved, and 20 kilograms of urea and 10 kilograms of phosphate fertilizer are applied per mu. The middle and later periods of the lotus base are key fertilization periods, furrowing and topdressing are carried out between rows or hole topdressing is carried out, 50 kilograms of compound fertilizer is applied per mu, and irrigation is carried out after furrowing is sealed. 15 kg of urea is applied to each mu in the early stage and the middle stage of the balling.

Thirdly, disease and pest control: the plant diseases and insect pests are less during the cultivation in spring, and mainly occur in the later growth stage. Biological agents and low-residual-toxicity pesticides are adopted to prevent and control pests such as cabbage caterpillars, aphids and plutella xylostella; meanwhile, yellow plates can be used for trapping and killing aphids.

4) Harvesting: the leaf bulbs are fully enlarged, the core is compact, and the bulbs should be harvested in time to prevent bulb cracking.

Test example 1

And (3) combination and matching: according to the parent matching principle of hybrid seeds in cabbage breeding in 2012, 8 cytoplasmic male sterile lines such as DH08-GW100CMS and the like are selected as female parents, 9 DH lines or self-bred lines with excellent economic properties, high matching force, strong winterness, crack resistance and strong disease resistance including DH12-3-M2 are selected as male parents, the matching force is researched and analyzed by a hybridization matching group, and tests in 2013-2014 in Yangling, Xian, Bao chicken and the like show that the combination of DH08-GW100CMS multiplied by DH12-3-M2 shows remarkable heterosis, excellent comprehensive economic properties, crack resistance, excellent planting, disease resistance and yield, meets the breeding target, and the results show that the yield reaches 100%, the low temperature resistance, the weight of a single bulb is about 1.2kg, the disease resistance and yield of a new variety are superior to those of a reference variety 8398 (the disease resistance table 1).

Table 1: comparison result of main target characters of kale Qinlian 1658

2. Innovation point of hybrid generation Qin Gan 1658 variety

2.1 the product has excellent quality and the green leaves.

By survey and analysis of the quality ratio in 2013-2015 and test and analysis of the quality supervision and inspection station for agricultural products in Shaanxi province, the Qinlian 1658 variety has excellent quality compared with the control 8398 variety, the spherical leaves are emerald green, the leaf quality is crisp and sweet, and the length of the central column is 5.1cm and is shorter than the control 0.8 cm; the compactness of the leaf ball is 0.7 which is greater than that of the contrast 0.11; the leaf ratio of the upper leaves is 20.9 percent and is less than that of the control by 3.4 percent. The nutritional quality analysis showed that the dry matter, soluble sugars, crude fiber and crude protein were all higher than the control, 1.1%, 0.47%, 0.27% and 0.25%, respectively (table 2).

TABLE 2 determination of the quality of Qinlian 1265 variety

2.2 stable and high yield and strong yielding ability.

The Qinlian 1658 variety is compared with a control 8398 variety, and the continuous 5-year high yield is outstanding through variety comparison test, regional test and production demonstration, and the average yield of the Qinlian 1658 variety is 3739.3kg/667m²And the yield is increased by 10.6 percent compared with a control 8398 variety.

(1) Sample ratio test: the results of the variety comparison test for three consecutive years from 2013 to 2015 are shown as follows: the average acre yield of the three-year Qin sweet 1658 variety is 3688.1kg/667m²The average yield per mu of the control variety 8398 is 3296.8kg/667m²And the yield of the Qinlan 1658 is increased by 11.8% compared with the control. (Table 1)

(2) Area test: the results of two-year continuous provincial region test in 2015-2016 show that the average acre yield of the two-year Qin-Gan 1658 variety is 3739.3kg/667m²The average yield per mu of the control variety 8398 is 3381.7kg/667m²And the yield of the Qinlan 1658 is increased by 10.6% compared with the control. In 2015, the Qin Gan 1658 variety is tested in a district, the yield is increased by 14.7% in a Hanzhong point, and is increased by 7.6% in a Taibai point, compared with a contrast, by a minimum; 2016The experiment of the Qin Gan 1658 variety in the district shows that the increase of the yield is the largest at the Yangling point and 13.8% at the contrast, and the increase of the yield is the smallest at the county and the county, and 6.5% at the contrast. (Table 3)

Table 3: yield results of the zone test for Qinlian 1658 variety

(3) Production demonstration: the result of the demonstrated yield is produced for two continuous years in 2015-2016, and the average yield per mu of Qinlian 1658 is 3543.2kg/667m²The average yield per mu of the control variety 8398 is 3295.6kg/667m²And the yield of the Qinlan 1658 is increased by 10.9% compared with the control. (Table 4)

TABLE 4 demonstration yield results for production of Qin Gan 1658 cultivar

2.3 resistant to cracking, high disease resistance and strong stress resistance.

Through field identification for many years, the ball breaking index is less than grade 3 after 5 days of late harvest, the ball breaking rate is less than 5 percent, and the ball is resistant to ball breaking; the winterness is strong, and the immature bolting phenomenon does not occur; and (5) identifying disease resistance in the field. High resistance to virus, black rot and dry heartburn. (Table 5)

TABLE 5 identification of disease resistance and stress resistance of Qin Ganshangan 1658 variety

2.4 the plant characters are neat and consistent.

The Qinlian 1658 variety is the first hybrid generation early-maturing spring cabbage variety prepared by using cabbage DH line as a substitute for inbred line as parents, and the high uniformity of DH line genotype enables the plant morphology, leaf color and nodulation of the Qinlian 1658 variety field to be highly regular.

3. Conclusion

Qinlian 1658(DH08-GW100CMS X DH12-3-M2) is a first generation variety of early-maturing hybrids of spring cabbage, and is planted until harvested for about 58 days; the cabbage cytoplasmic male sterile line DH08-GW100CMS and DH line DH12-3-M2 are hybridized and bred, parents are double haploid inbred lines (DH lines), the first hybrid generation field performance uniformity is high, plant types are semi-upright, outer leaves are green, leaf surfaces are smooth, wax powder is less, and leaf bulbs are emerald green; the plant expansion degree is 38.2-39.1 cm, the number of outer leaves is 10-11, the weight of a single sphere is about 1.2kg, the leaf sphere is circular, the compactness of the leaf sphere is 0.70, the length of a central column is 5.1cm, the height of the central column is 1/2% smaller than the sphere, and the leaf ratio is 20.9%. High resistance to virus, black rot and dry heart, high resistance to cracking and high winter performance. The leaf balls are round and regular, the marketability is good, and the appearance of the leaf balls accords with the habit of consumers; the ball leaves are crisp and sweet in texture and excellent in flavor quality. The variety has high and stable yield, the average yield is 3739.3kg/667m2, and the yield is increased by 14.3 percent compared with that of a control 8398 variety; is a new high-quality, disease-resistant and high-yield spring cabbage cracking-resistant bulb variety.

The genetic resources DH08-GW100M3CMS, Ogura, DH08-GW100M3 and DH12-3-M2 related to the invention are all from the seed bank of the gardening college of northwest agriculture and forestry science and technology university.

Claims (1)

1. A method for breeding a new early-maturing variety by utilizing CMS and DH lines of cabbage is characterized by comprising the following steps:

1) breeding of female parent germplasm material DH08-GW100M3CMS

The female parent DH08-GW100M3CMS cytoplasmic male sterile line is formed by hybridizing an Ogura cytoplasmic cabbage male sterile line serving as a female parent material with a cabbage excellent DH08-GW100M3 line and then carrying out 5-generation backcross transformation;

the female parent maintenance line DH08-GW100M3 is a crack-resistant ball excellent double-haploid system, namely a DH line, which is created by culturing microspores of cabbage early-maturing hybrid expected through a cabbage isolated microspore culture technology;

2) breeding of male parent germplasm material DH12-3-M2

The male parent DH12-3-M2 is a disease-resistant high-quality DH12-3-M2 obtained by culturing a cabbage early-maturing variety Qinlian 50 through free microspores and identifying through selfing;

3) selection of hybrid seeds

A cabbage cytoplasmic male sterile line DH08-GW100M3CMS is selected as a female parent and a DH line DH12-3-M2 is selected as a male parent to be hybridized and bred into a DH08-GW100M3CMS xDH 12-3-M2 combination which is named as Qinlian 1658 variety.

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