CN106416995A - Method for breeding novel variety of Brassica oleracea by using two DH (dihaploid) lines - Google Patents

Abstract

The invention discloses a method for breeding novel variety of Brassica oleracea by using two DH (dihaploid) lines, comprising: hybridizing a cytoplasmic male-sterile line DHCMS08-5-3-6-9 trans-bred with a Brassica oleracea DH line as a female parent and the Brassica oleracea DH line DH05X-1-5-9-8 line as a male parent to prepare DHCMS08-5-5-6-9*DH05X-1-5-9-8 hybrid first-generation combination, named as Qingan 1562. The method of the invention is a method to breed a mid-early maturity novel variety of Brassica oleracea by using two DH lines, a novel hybrid first-generation variety of Brassica oleracea is bred through DH liens with genotype purity of 100% created by culturing isolated microspores of Brassica oleracea, the novel hybrid first-generation variety of Brassica oleracea has highly uniform economic traits and completely exhibited target traits, and the breeding of disease-resistant high-quality high-yield variety breeding targets is achieved.

Description

Utilize the method for two DH systems, one wild cabbage new varieties of seed selection

Technical field

The invention belongs to Crop Genetics and Breeding section in agricultural science and technology, be specifically related to one and utilize two DH systems seed selection one The method of wild cabbage new varieties.

Background technology

Wild cabbage breeding of new variety is that the vegetable crop breed breeding established for a long time of country is mainly tackled key problems research project One of.Wild cabbage hybrid vigour is obvious, and the new varieties cultivated now are half-blood；Elite inbred, cytoplasm male sterility line Screening and heterosis utilization have become one of effective way cultivating multiple target new varieties.The collocation of wild cabbage new varieties is planted in agriculture Industry industry restructuring, increase farmers' income on play a leading role.Head cabbage varieties many batches listings in many seasons, to alleviating, market is light In the busy season, meet the stable market supply of fresh vegetable anniversary and play key effect；Wild cabbage is rich in nutriment and medical value, to enhancing mankind's body Body is healthy and increase food dish designs and varieties of uplifting the people's living standard play extremely important effect.Virus in current China Cabbage production Sick, black rot is on the rise, and old variety deterioration, horizontal resistance difference can not efficiently solve production problem, and shoddy product is low, aggravation The demand and supply contraction；The specific breed that wild cabbage ball leaf is green, leaf is thick, dry matter content is high lacks, and stops current industry development；City Field not only needs how anti-kind, and proposes requirements at the higher level to quality.Wild cabbage is to virosis（TuMV、CMV）And black rot（Br）'s Breeding for disease resistance becomes the most effective approach mitigating disease.The high material containing Major Nutrient of wild cabbage, eating mouth feel crisp sweet with Crude fibre is less becomes again consumer demand.Thus, current wild cabbage resists two or more disease, and high-quality, high yield, resistance to blastomere just become The main breeding objective of Mid-late ripening wild cabbage.

Traditionally wild cabbage first generation of hybrid kind preparation is all to select two through many self-mating systems separating acquisition for artificial autocopulation Hybridization forms, and the character gene of self-mating system isozygotys limitation, and hybridization generation proterties regularity is relatively poor, is bred as new product The objective trait performance planted is insufficient.

Content of the invention

Utilize for routine two self-mating system hybridization generation kind technology exist self-mating system easily degenerate, hybrid neat Degree difference and hybrid vigour can not the fully problems such as performance and defect, it is an object of the invention to provide one and utilize two DH Being the method for one wild cabbage new varieties of seed selection, its DH system individual plant selfing algebraically of head cabbage varieties being bred as through the method is few, growth potential By force, first generation of hybrid breeding seed yield is high, and two DH system hybridization generation regularities of high homogenous are high, its improved variety warp Comparative test, regional experiment of variety and production demonstration, performance hybrid vigour is notable, and between hybrid plant, proterties presents completely the same, Integral economic trait is excellent, and yielding ability highlights, and varietal adaptation is strong, and volume increase advantage is stable, and by state verification name, " Qin is sweet 1562 " head cabbage varieties.

To achieve these goals, the technical solution used in the present invention is to utilize one wild cabbage mid-early maturity of two DH systems seed selection The method of kind, comprises the following steps：

1）The seed selection of maternal germplasm materials DHCMS08-5-3-6-9

（1）The seed selection of DH08-5-3-6-9

DH08-5-3-6-9 system is to utilize two wild cabbage self-mating systems MP01-2566 × Y03-3621 to prepare F1, through to its sporidiole Cultivating the excellent DH system being bred as with field test, economical character is highly ordered, disease-resistant, high-quality, and coordinate force is high；

（2）DHCMS08-5-3-6-9 transformation

DHCMS08-5-3-6-9 cytoplasm male sterility line is that the wild cabbage first generation of hybrid that utilization has Ougra male sterility gene is Female parent, with DH08-5-3-6-9 system for the male parent that backcrosses, hybridizes through 5 generations and backcrosses incubation DHCMS08-5-3-6-9 cytoplasmic male not Educate and be；

2）The seed selection of male parent germplasm materials DH05X-1-5-9-8

Male parent DH05X-1-5-9-8 system is " to be wished " by mid-early maturity wild cabbage first generation of hybrid kind, separates and orientation choosing through selfing It after educating the excellent three-generation inbred line of acquisition, then to its three-generation inbred line through microspores culture and field Characters Identification, is bred as gene pure Close the excellent DH system of High quality and diseases resistance；

3）The first generation of hybrid combines seed selection

The cytoplasm male sterility line DHCMS08-5-3-6-9 becoming with the transformation of wild cabbage DH system for maternal with the wild cabbage DH i.e. DH05X-1-of system 5-9-8 system is male parent, is configured to the combination of DHCMS08-5-3-6-9 × DH05X-1-5-9-8 first generation of hybrid, name by hybridization For the Qin sweet 1562.

The Qin sweet 1562 is through 2 years variety tests and regional experiment of variety and produces demonstration, and integral economic trait is excellent Good, show disease-resistant harvesting high-quality high-yield, objective trait highlights and significantly beyond check variety.

The method of the present invention is to select the DH system isozygotied by wild cabbage microspore-isolated culture initiative genotype 100% to be bred as The new first generation of hybrid kind of one wild cabbage, its variety economic character presents highly ordered, and objective trait shows completely, it is achieved disease-resistant excellent The seed selection success of matter high yield breeding for quality target.

Brief description

The maternal DHCMS08-5-3-6-9 cytoplasm male sterility line seed selection figure of Fig. 1；

Fig. 2 male parent DH05X-1-5-9-8 excellent DH system seed selection figure；

The DH strain figure of the DH08-5-3-6-9 of Fig. 3 robust growth；

The DH system figure of Fig. 4 proterties clean fit power height DH08-5-3-6-9；

Fig. 5 DHCMS08-5-3-6-9 system floral organ figure；

The DH strain figure of Fig. 6 proterties clean fit power height DH05X-1-5-9-8；

The DH system figure of Fig. 7 proterties clean fit power height DH05X-1-5-9-8；

Sweet 1562 kinds of Fig. 8 wild cabbage disease-resistant harvesting high-quality high-yield mid-early maturity Qin（DHCMS08-5-3-6-9×

DH05X-1-5-9-8）Figure.

Detailed description of the invention

The specific embodiment being given below by way of inventor and concrete regional testing example beneficial effects of the present invention is made into The detailed description of one step.

Embodiment 1

1. two the DH system parent's seed selections of sweet 1562 first generation of hybrid kind of the Qin and trait expression

1）The seed selection of maternal germplasm materials DHCMS08-5-3-6-9

（1）The seed selection of DH08-5-3-6-9

The holding that Fig. 1 gives maternal DHCMS08-5-3-6-9 cytoplasm male sterility line is seed selection figure, and maternal holding is DH08-5- 3-6-9 system is two the wild cabbage self-mating system MP01-2566 × Y03-utilizing the seed selection of College of Horticulture of western agriculture and forestry science and technology university for 2007 3621 preparation F1, through to its F1 microspores culture（Fig. 3 DH08-5-3-6-9DH strain）The excellent DH being bred as with field test System（The DH system of Fig. 4 DH08-5-3-6-9）, economical character is highly ordered, disease-resistant and high-quality（Table 1）, coordinate force is high.

（2）DHCMS08-5-3-6-9 transformation

Fig. 1 gives the seed selection figure of maternal DHCMS08-5-3-6-9 cytoplasm male sterility line, DHCMS08-5-3-6-9 cytoplasmic male Sterile line be the 2008 annual interests wild cabbage first generation of hybrid with Ougra male sterility gene be maternal, with DH08-5-3-6-9 system For the male parent that backcrosses, hybridize through 5 generations and backcross in incubation DHCMS08-5-3-6-9 cytoplasm male sterility line in 2012（Fig. 5 DHCMS08-5-3-6-9 system sterility floral organ）.

（3）Maternal owner wants trait expression

DH08-5-3-6-9 system and DHCMS08-5-3-6-9 cytoplasm male sterility line in addition to fertility is inconsistent, other trait expression Unanimously.Sterile line and holding are that karyogene proterties is isozygotied, and plant strain growth is healthy and strong, bloom normal, and nectary is flourishing, and gynoecium is normal； The stamen development of DH08-5-3-6-9 system is normal, and DHCMS08-5-3-6-9 cytoplasm male sterility line staminody, sterility are thorough； Sterile line and holding are ripe in ripe property, ripe 65 days of field planting to leaf-head；Plant lotus throne leaf half is upright, and leaf look green, in wax powder, and leaf Ball is round；Best in quality, newel 5.2cm, the degree of packing 0.61；High anti-TuMV, CMV, disease refers to≤1.34, and black rot resistance, disease refers to 3.33（Table 1）；Planting strain branchiness relatively strong, spray growing way is strong, average kind plant height degree 102cm, florescence 30～35d.

2）The seed selection of male parent germplasm materials DH05X-1-5-9-8

（1）The seed selection of DH05X-1-5-9-8

Fig. 2 gives male parent DH05X-1-5-9-8 excellent DH system seed selection figure, male parent DH05X-1-5-9-8 system be 2000 by state One High quality and diseases resistance mid-early maturity wild cabbage first generation of hybrid kind of outer introducing " is wished ", separates and fixed through selfing in 2002～2005 years After seed selection obtains excellent three-generation inbred line, then to its three-generation inbred line through 2006～2007 years microspores culture（Fig. 6 DH05X-1-5-9-8DH strain）It with field Characters Identification, is bred as the excellent DH system of gene pure High quality and diseases resistance（Fig. 7 DH05X-1- 5-9-8DH system）.

（2）Male parent owner wants trait expression

The ripe sex premature of male parent system DH05X-1-5-9-8, ripe 55 days of field planting to leaf-head；Plant siphonal lobe is more, and strain state is closely upright, leaf Look yellow green, wax powder is few, and leaf-head is circular, leaf-head pole consolidation, the degree of packing 0.69；Best in quality, blade is crisp sweet, newel 5.0cm, Side leaf ratio 18.5%；High antiviral is sick（TuMV disease refers to that the 1.67th, CMV disease refers to 0.67）With high black rot resistance（Disease refers to 1.67）（Table 1）； Planting strain robust growth, blooming normal, stamen and gynoecium are normal, in spray branchiness, in spray growing way, averagely plant plant height 98.5cm, florescence 30～35d.

3）The first generation of hybrid combines seed selection

The cytoplasm male sterility line DHCMS08-5-3-6-9 that seed selection in 2012 becomes with the transformation of wild cabbage DH system for maternal with wild cabbage DH system I.e. DH05X-1-5-9-8 system is male parent, is configured to DHCMS08-5-3-6-9 × DH05X-1-5-9-8 first generation of hybrid by hybridization Combination（Sweet 1562 kinds of Fig. 8 Qin（DHCMS08-5-3-6-9×DH05X-1-5-9-8））, this combination was through 2 years variety protection Test and regional experiment of variety and production are demonstrated, and integral economic trait is excellent, shows disease-resistant harvesting high-quality high-yield（Table 1）, objective trait Highlight and significantly beyond check variety, the named Qin sweet 1562.

Table 1：Two DH systems and first generation of hybrid disease resistance and quality show

4）The feature of sweet 1562 kinds of the Qin

Sweet 1562 kinds of the Qin are first cross kind, Early-medium maturing variety, gather in the crops 62 days from field planting to leaf-head.Plant is relatively upright, plant height 35.2cm, degree of development 46.2 cm × 45.1cm, the outer number of sheets 15, siphonal lobe yellow green, in wax powder, leaf-head is round, ball leaf green； Ball indulges footpath 15.2cm, transverse diameter 15.5cm；Leaf-head core length 5.2cm, single ball weight 1.5kg；The leaf-head degree of packing 0.62, quality is excellent Good, the food value of leaf is crisp sweet, and commodity is good, resistance to blastomere；Viral diseases and black rot resistance；Per mu yield level 5000～5500kg.

2. the technical essential that sweet 1562 hybrid seeds of the Qin produce

Half-blood seed takes half strain breeding, selects the area that suitable brassica specie strain grows, seed farm ensure with other ten Seed farm interval more than the 2km of Zi Hua section brassica plant and wild cabbage class different cultivars.Guanzhong Region, Shaanxi Province, China dew on general August 5 Ground nursery, open country field planting on September 15.Parents' field planting ratio is female parent（DHCMS08-5-3-6-9）：Male parent（DH05X-1-5-9- 8）=31, line-spacing 55～60cm, spacing in the rows 35～40cm, male parent spacing in the rows can rational close planted.Parents survive the winter before strengthen root system ridging and Winter irrigation, plants built after strain bolting, preventing and treating plant lodging；Florescence bee pollination, seed farm puts honeybee 1～2 case/mu, after planting strain the flowers are in blossom, Forbid spraying medicine insect protected, in order to avoid killing bee colony, the maternal florescence terminate after pull out male parent system in time, after seed harvest, timely airing, anti- Only go mouldy.

3. the sweet 1562 variety culture technical essentials of the Qin

1）Seeding and seedling raising:

The Northwest's outdoor cropping in spring, can be in mid or late December cold bed seeding and seedling raising or early and middle ten days in January greenhouse or improvement cold bed Seeding and seedling raising, sticks together field planting with band soil during 6～7 true leaves of young plant during field planting.

2）Field planting：

Outdoor cropping is after early spring, soil thawed completely, and temperature is constant more than 10 DEG C, field planting when young plant has 6～7 true leaves.Spring During season young plant field planting, temperature is relatively low, suitably uses seat water to plant seedling, favorably improves ground temperature.Use flat furrow or half high bedding cultivation, density 4000 seedling/667；Otherwise cultivate overstocked or excessively dilute, yield all can be caused to reduce.

3）Field management：

1. pour water：Field planting is poured water once after delaying seedling, and at this moment temperature and ground temperature are relatively low, and irrigation quantity is unsuitable excessive.Control subsequently Pour water or because soil overdrying can little be poured water once, until the balling initial stage is arrived in restraining the growth of seedlings.Treating Bao Xinhou, temperature raises, and need to strengthen and pour water Amount, increases frequency of irrigation；Meet personally and dry see wet, just should pour water when namely meeting personally dry.Pour water altogether growth period 4～5 times, adopting Suitably to control moisture before receipts, prevent blastomere, affect yield and product quality.

2. apply fertilizer：Topdress 3～4 times.Based on early stage of topdressing is with nitrogen, phosphorus, potassium, based on the later stage is with nitrogenous fertilizer.Heavy dressing base manure, mu is executed Fertilizer 5000kg, composite fertilizer 50kg；Intertillage in about 10 days after field planting is topdressed once, and mu imposes l5 kilogram of urea, provides lotus throne The adequate nutrients of leaf growth.Entering the lotus throne initial stage, spreading manuer in holes and topdress, improve fertilising concentration, mu imposes 20 kilograms of urea and phosphate fertilizer 10 Kilogram.The lotus throne middle and later periods is emphasis fertilizer application period, and in the ranks trench digging topdresses or spreads manuer in holes and topdresses, and mu imposes composite fertilizer 50 kilograms, after envelope ditch Pour water.Balling initial stage and mid-term, mu respectively imposed 15 kilograms of urea.

3. insect pest preventing and controlling：Spring cultivation pest and disease damage is less, occurs mainly in Later growth.Use biological agent and low residual hazard The insects such as pesticide control cabbage caterpillar, aphid and diamond-back moth；It is possible with yellow card's trap aphid simultaneously.

4）Gather：

Leaf-head is fully loose, bag heart consolidation, should gather in time, prevent blastomere.

Test example 1：Sweet 1562 variety tests of the Qin and production demonstration（Shaanxi）

Wild cabbage Early-medium maturing variety combination selection research in 2011～2013 years, selects with DHCMS08-5-3-6-9 cytoplasmic male sterilty System is female parent, with the high DH system of 9 economic characters such as DH05X-1-5-9-8 TuMV excellent, anti-, Br, CMV, coordinate force as male parent, Coordinate force is researched and analysed, through 3 years in College of Horticulture of Xibei Univ. of Agricultural ＆ Forest Science ＆ Technology experimental plot by hybridization combo（Yang Ling）、 The ground such as Xi'an, Baoji carries out variety test and produces demonstration（Table 2）.Result shows DHCMS08-5-3-6-9 × DH05X- 1-5-9-8 combination performance hybrid vigour is notable, and integral economic trait is excellent, and disease resistance, quality and yielding ability highlight；Meet new Breeding for quality target, its major economic indicators be mid-early maturity, field planting to results 62 days, best in quality, the food value of leaf is crisp sweet, commodity Good, leaf-head ball-shaped, newel 5.2cm（High less than ball by 1/2）, leaf-head consolidation 0.62, receive 7 days evening without blastomere rate, average yield per mu Amount 5249.7kg, sweet 21 volume increase 5.4 % than in comparison.High antiviral is sick, and black rot, and disease index is respectively 0.67 He 1.95.

Sweet 1562 variety tests of table 2 Qin and production demonstration yield result（Shaanxi）

Test example 2：Sweet 1562 regional experiment of variety of the Qin and production demonstration（The whole nation）

Sweet 1562 kind of the Qin is participated in whole nation head cabbage varieties regional testing for 2013～2015 years and produces demonstration, and this kind commodity are ripe Average 63.4 days of phase, belong to Early-medium maturing variety.Plant strain growth gesture is strong, dark green leaf color, in wax powder, and leaf-head ball-shaped, compared with consolidation 0.62, Core length accounts for high 0.47 of ball, average single ball weight 1.50kg（Table 3）, resistance to blastomere is good, is significantly better than comparison.3 years variety plots Territory test and production demonstration per mu yield 5369.5kg, than in comparison, sweet 21 volume increase 8.5%, occupy tested variety the 2nd.

The sweet 1562 kind principal character characteristics of table 3 whole nation wild cabbage regional testing and the production demonstration Qin

Regional testing in 2013：In addition to the yield in Beijing and Qinghai district pilot is relatively low, other point be above comparison in sweet No. 21,8 The overall average per mu yield of individual district pilot is 5345.5kg, ratio comparison volume increase 5.7%, occupies second in 9 tested varieties（Table 4）.

Regional testing in 2014：Sweet 21,8 districts examination higher than in comparison of the yield of 7 district's pilots in addition to Shanxi district pilot The overall average per mu yield of point is 5493.0kg, ratio comparison volume increase 13.6%, occupies second in 9 tested varieties（Table 4）.

Table 4 whole nation wild cabbage regional testing tested variety yield（Unit：kg/667m2）²

Variety name	Regional testing in 2013	Regional testing in 2014	Average product	Precedence
					Sweet No. 1 of Henan	5228.5	4879.0	4993.0	5
06-103	4956.5	4570.0	4581.1	9
					The Qin sweet 1562	5345.5	5493.0	5419.3	2
06-28	5074.2	4789.0	4725.8	8
					Sweet No. 4 of Henan	5229.2	5127.0	4875.8	7
06-192	5187.2	5293.0	5240.1	4
					Rich No. 6 of favour	5236.9	5429.0	5258.6	3
06-196	5829.3	5716.0	5484.7	1
					In sweet No. 21 (CK)	5059.7	4837.7	4948.7	6

Within 2014, produce demonstration and be shown in Table 5：

Table 5 whole nation Cabbage production test tested variety yield（Unit：kg/667m²）

The yield of 6 district's pilots in addition to Beijing and Hebei higher than the overall average per mu yield of sweet 21,8 district's pilots in comparison is 4976.6kg, ratio comparison volume increase 5.6%, in 9 tested varieties, occupy second.

Claims (1)

1. utilize the method for two DH systems, one wild cabbage new varieties of seed selection, it is characterised in that include the following step：

1）The seed selection of maternal germplasm materials DHCMS08-5-3-6-9

（1）The seed selection of DH08-5-3-6-9

DH08-5-3-6-9 system is to utilize two wild cabbage self-mating systems MP01-2566 × Y03-3621 to prepare F1, through microspores culture The excellent DH system being bred as with field test, economical character is highly ordered, disease-resistant, high-quality, and coordinate force is high；

（2）DHCMS08-5-3-6-9 transformation

DHCMS08-5-3-6-9 cytoplasm male sterility line is that the wild cabbage first generation of hybrid that utilization has Ougra male sterility gene is Female parent, with DH08-5-3-6-9 system for the male parent that backcrosses, hybridizes through 5 generations and backcrosses incubation DHCMS08-5-3-6-9 cytoplasmic male not Educate and be；

2）The seed selection of male parent germplasm materials DH05X-1-5-9-8

Male parent DH05X-1-5-9-8 system is " to be wished " by mid-early maturity wild cabbage first generation of hybrid kind, separates and orientation choosing through selfing It after educating the excellent three-generation inbred line of acquisition, then to its three-generation inbred line through microspores culture and field Characters Identification, is bred as gene pure Close the excellent DH system of High quality and diseases resistance；

3）The first generation of hybrid combines seed selection

The cytoplasm male sterility line DHCMS08-5-3-6-9 becoming with the transformation of wild cabbage DH system for maternal with the wild cabbage DH i.e. DH05X-1-of system 5-9-8 system is male parent, is configured to the combination of DHCMS08-5-3-6-9 × DH05X-1-5-9-8 first generation of hybrid, name by hybridization For the Qin sweet 1562.