CN103444515A - Transfer method for male sterile line of purple wuta-tsai - Google Patents

Abstract

The invention discloses a transfer method for a male sterile line of purple wuta-tsai. In the traditional method, an Ogura sterile source has the defects that plants are yellowed when meeting a low temperature at a seedling stage, a floral organ nectary is degenerated and the like; in the method disclosed by the invention, a high-quality Ogura cytoplasm male sterile source purple non-heading Chinese cabbage male sterile line RC1 is utilized to be hybridized with a purple wuta-tsai stable selfing line PW1501; the PW1501 is used as a backcrossing male parent to continuously backcross and transfer for six generations; a wuta-tsai Ogura cytoplasm male sterile line RHP16 which is stable in sterility and has sterile plant rate and sterile degree being up to 100% is bred through economic character selecting and disease resistance and stress resistance identifying. According to the sterile line, the plants are not yellowed at the low temperature during the seedling stage, the nectary is developed, the sterility is stable, the combining ability is good, the comprehensive resistance is strong and the utilization value is relatively high.

Description

A kind of transfer method of purple Wuta-tsai male sterile line

Technical field

The invention belongs to the brassicaceous vegetable breeding technical field, particularly, the invention belongs to a kind of invention of purple Wuta-tsai male sterile line transfer method.

Background technology

Wuta-tsai is the mutation that the Cruciferae rape belongs to Vegetable Crops of Brassica Chinese cabbage subspecies, originates from basin, China middle and lower reach of Yangtze River, now the cultivation history of existing nearly one thousand years.Wuta-tsai adaptability is wider, and current national most area all has cultivation, and some places are as the special vegetable introduction of plant.Wherein south cultivation is more, especially in Anhui, the ground such as Shanghai, Jiangsu eat very general, area under cultivation is larger.The Wuta-tsai tolerance to cold is strong, especially is applicable to planted in fall, and supply around the Spring Festival, with sweet and famous through the frost and snow aftertaste, is to adjust the desirable vegetables of " winter lacks " and " spring is light ", just as important vegetables, cultivates since ancient times.Wuta-tsai not only is subject to domestic consumer's favor deeply, and has been subject to external attention, and export volume constantly increases.But, the research of Wuta-tsai is rested on substantially to the economical character evaluation of external form, in Variety Introduction and culture technique research.The position of work in extreme backwardness of Wuta-tsai breeding aspect, the Wuta-tsai of production use mostly is conventional and plants, and has restricted greatly the development of Wuta-tsai industry.Research shows, the hybrid vigour of Wuta-tsai is very remarkable, but lacks optimized production of hybrid seeds new technology, and its heterosis utilization is limited by very large.

Male sterile is ubiquitous a kind of phenomenon in higher plant.Cruciferae Brassica Crops hybrid vigour is obvious, is the ideal material that utilizes male sterile line to carry out hybrid seed production, and the seed selection of crossbreed has become breeding method and the trend that this owner wants.Utilize male sterile line to be cross-breeding and not only can save the labour of artificial emasculation, can also avoid the parent to degenerate because of the merit that selfing causes continuously, obtain success greatly in crossbreeding, therefore, based on male sterile crossbreeding, also become a kind of efficient breeding method.

Male sterile source the earliest is the little storehouse of Ogura(, 1968) in certain radish varieties seed field of Japan, find, and through this sterility of evidence by plasmon and 2 pairs of adelomorphic cell nuclear gene co-controllings, this sterile source is called Ogu CMS, and it is also current most study, utilization one of cytoplasmic male sterility source the most widely.The advantage in Ogura male sterile with radish cytoplasm source is that staminody is thorough, and without pollen, sterile plant rate and sterile degree are 100%, but exists blade yellow, growth retardation, floral organ nectary to degenerate and the defect such as coordinate force is poor.The Pelletier of nineteen eighty-three France academy of agricultural sciences etc. improves Ogu CMS by protoplast fusion, obtained the sterile plant of the green of acting normally under low temperature, Normal nectary.Afterwards, Chinese scholars is by Asymmetric Protoplast Fusion, hybridization, how for technique improvements such as backcross transformations, now successfully be bred as the not yellow of low temperature plant, the nectary developments such as cauliflower, Chinese cabbage, wild cabbage, purple tsai-tai cytoplasm male sterility line preferably, but transformation purple Wuta-tsai male sterile line there is not yet report.

Summary of the invention

Technical problem to be solved by this invention is for the deficiencies in the prior art, a kind of transfer method of purple Wuta-tsai male sterile line is provided, can select sterility by the method stable, sterile plant rate and sterile degree reach 100%, not yellow of low temperature plant in seedling stage, the nectary prosperity, sterility is stable, the purple Wuta-tsai Ogura cytoplasm male sterility line kind that coordinate force is good.

Technical solution problem of the present invention adopts following technical scheme:

A kind of purple Wuta-tsai male sterile line transfer method, the method be take Chinese cabbage Ogura cytoplasmic male sterilty source purple Chinese cabbage male sterile line RC1 as maternal, stablize inbred line PW1501 hybridization with the purple Wuta-tsai, and with inbred line PW1501 for the male parent that backcrosses continuous 6 generation backcross transformation, select sterility stable, sterile plant rate and sterile degree reach 100% Wuta-tsai Ogura cytoplasm male sterility line.

Described Wuta-tsai is stablized inbred line PW1501 seed selection acquisition as follows:

1) select the purple Chinese cabbage PC601 of inheritance stability;

2) select the good Wuta-tsai inbred line of Comprehensive Traits;

3) take purple Chinese cabbage PC601 as maternal, is that male parent is hybridized with the Wuta-tsai selfing, obtains the F1 plant;

4) select purple F1 plant to do maternal, backcross with the Wuta-tsai inbred line, obtain the BC1 seed;

5) do maternally with the purple plant of BC1 seed, with Wuta-tsai inbred line continuous backcross 3-6 time, obtain BCn plant identical with paternal character, the blade purple;

6) purple BCn plant is carried out to selfing and obtain seed BCnS1;

7) BCnS1 is selected to the Wuta-tsai plant that acquisition inheritance stability, leaf look are purple through conventional field test combination;

8) the purple Wuta-tsai plant of acquisition is continued to the Wuta-tsai seed that a large amount of purples of numerous acquisition are expanded in selfing.

Described Wuta-tsai inbred line is selected from YW905.

Purple Chinese cabbage PC601 in the present invention and two kinds of germplasm resources of Wuta-tsai inbred line YW905 all are kept in the Horticultural Research Institute species bank of Academy of Agri-Science and Technology Anhui Province.Wherein the purple Chinese cabbage derives from the purple Chinese cabbage variant of finding in the Horticultural Research Institute variety source of Academy of Agri-Science and Technology Anhui Province, by preferably involving continuous individual plant selfing purifying in a criminal case, the a series of deep mixed strain obtained, select the more gorgeous further selfing purifying of strain of color, obtain purple Chinese cabbage that the offspring separates without the proterties inbred line PC601 that isozygotys; Wuta-tsai inbred line YW905 is that the Anhui Native Wuta-tsai resource six that will collect is pacified the gold zone crows by the selfing continuously of 3-5 generation, the inbred line of isozygotying of the stabilization characteristics of genetics of acquisition.

The not yellow of male sterile line low temperature in seedling stage plant of the inventive method transformation; Sterility is stable, and nectary development is good, and column cap is normal, and blossoming and bearing fruit property is good; Coordinate force is good; Comprehensive resistance is strong, is good male sterile line.

The accompanying drawing explanation

The transformation family tree that Fig. 1 is purple Wuta-tsai cytoplasm male sterility line.

Embodiment

Below in conjunction with embodiment and accompanying drawing, the present invention is described in detail.

The parent selects: Ogura cytoplasmic male sterilty source RC1: buy and obtain, and the face of blade purple, the leaf back green, petiole is emerald green, and its sterile degree, sterile plant rate are 100%, and seedling stage is without aetiolation.The male parent that backcrosses inbred line PW1501(PW1501 is Academy of Agri-Science and Technology Anhui Province Horticultural Research Institute Germplasm Bank resource), PW1501 cultivates acquisition in the following manner: (PC601 is Academy of Agri-Science and Technology Anhui Province Horticultural Research Institute Germplasm Bank resource as maternal to take the PC601 of purple Chinese cabbage variant selfing purifying, from the Academy of Agri-Science and Technology Anhui Province Horticultural Research Institute, experimental field find), take from the local germplasm of gold zone crow of Liuan City, Anhui Province Yuan District collection---the six peace gold zone crow inbred line YW905 that six peace gold zone crows obtain after the inbreeding of more generation purifying are male parent (YW905 is Academy of Agri-Science and Technology Anhui Province Horticultural Research Institute Germplasm Bank resource), hybridize and obtain purple F1 plant, the YW905 of take again backcrosses as male parent, (identify its leaf look in conjunction with field test, leaf, petiole, plant type, whether output etc. are similar to the male parent that backcrosses) the selection purple plant, through continuous backcross, obtain the Wuta-tsai backcross progeny of purple, obtain by selfing the purple Wuta-tsai inbred line PW1501 that hereditary capacity is stable again.This strain plant half is upright, plant height 18-20cm, and degree of development 22-25cm, the blade subcircular, there is the blister gauffer on blade face, siphonal lobe purple, petiole light green color.Cold-resistant, disease resistance is strong, and fiber is few, quality tender (table 1).

The disease-resistant resistance of table 1 different cultivars relatively

The backcross transformation of sterile material: select that growth potential in purple Chinese cabbage male sterile line RC1 plant is strong, Normal nectary, without losing complete sterility individual plant that flower bud, cross-fertile rate are good for maternal, the purple Wuta-tsai of take is stablized inbred line PW1501 as male parent, adopts artificial bud pollination technology to hybridize and obtains the F1 plant.Artificial bud pollination method: male parent was got pollen at bagging the previous day of blooming (removing open flower) the same day of blooming, and maternal flower bud phase stripping flower bud, be applied in fresh paternal pollen on maternal column cap, obtains transformation generation seed F1.

Autumn and winter and investigate the botany proterties of F1 plant spring in next year then, the growth potential of selecting and remain is strong, Normal nectary, cold-resistant, disease-resistant, similar to the transformation male parent without the fine individual plant work female parent that loses flower bud, take purple Wuta-tsai inbred line PW1501 as male parent backcrosses, obtain first backcross generation seed BC1.

Cold-resistant, Resistance screening method is: strain phase investigation plant, be subject to freeze injury degree and plant downy mildew, the virus disease incidence of disease.

Cold-resistant index=∑ (the cold-resistant rank of k*) * 100/5* investigates total strain number; K is appropriate level strain number.

Cold-resistant grade scale:

0 grade, manifest symptom, do not appear in the plant normal growth;

1 grade, leaf color jade green, a small amount of (1) siphonal lobe is slightly wilted;

2 grades, 2-3 sheet siphonal lobe is slightly wilted;

3 grades, 3 above outer leafs are wilted;

4 grades, internal lobe is softening wilts, but can recover;

5 grades, the softening wilting inside arranged, irrecoverable.

Cold-resistant evaluation criterion: cold-resistant: cold-resistant index (0-5): in anti-: cold-resistant index (5-10): cold-resistant: cold-resistant index (10-20): sensitivity: cold-resistant index (20-100).

The BC1 seed of acquisition is carried out to field test relatively, according to plant type, resistance, the proterties breeding goals such as output, select growth potential strong, Normal nectary, cold-resistant, disease-resistant, the nothing similar to the transformation male parent loses the backcross female parent of the fine individual plant of flower bud as next round, the purple Wuta-tsai inbred line PW1501 of take is the male parent continuous backcross, by field selection and continuous 6 generations backcross transformation acquisition sterility are stable repeatedly, sterile plant rate and sterile degree reach 100%, not yellow of low temperature plant in seedling stage, the nectary prosperity, coordinate force is good, disease-resistant, cold-resistant purple Wuta-tsai Ogura cytoplasm male sterility line RHP16(is now Academy of Agri-Science and Technology Anhui Province Horticultural Research Institute Germplasm Bank resource).

The principal character characteristic of purple Wuta-tsai male sterile line:

The floral organ feature: flower pesticide is shrivelled, white, fine, be degenerated to sarconeme shape, examines under a microscope, and can not find the pollen of this male sterile line; The many mastoid processes of stigma surface, ovary mode of appearance size is basic identical with maintainer, and structure function is normal; Nectary development is good, can attract the normal gathering honey pollination of insect such as honeybee.

The sterility performance: be not subject to the impact of environmental condition, sterility is stable, sterile plant rate 100%, sterile degree 100%.

Comprehensive Horticultural Characters: through too much, for backcross transformation and selection, the purple Wuta-tsai male sterile line RHP16 of incubation is basically identical with corresponding maintainer PW1501 main plant proterties.Plant half is upright, plant height 15cm left and right, and degree of development 28cm * 28cm left and right, the blade subcircular, there is the blister gauffer on blade face, siphonal lobe purple, the green white of petiole, the heavy 265g of individual plant left and right.Cold-resistant, disease resistance is strong, and fiber is few, quality tender (table 2).

The comprehensive Horticultural Characters of table 2 purple Wuta-tsai male sterile line RHP16 and maintainer relatively

Fecundity: male sterile line RHP16 and maintainer PW1501 thereof are in naturally solid test, and fecundity is in Table 3.Set seeds number and thousand kernel weight of the mono-pod of RHP16 is respectively 18.9 and 2.36g, than corresponding maintainer PW1501, distinguishes many 1.1 and 0.61g, increases respectively 6.2% and 34.8%.

The fecundity of table 3 purple Wuta-tsai male sterile line RHP16 and maintainer relatively

Claims (3)

1. a purple Wuta-tsai male sterile line transfer method, it is characterized in that: the method be take Chinese cabbage Ogura cytoplasmic male sterilty source purple Chinese cabbage male sterile line RC1 as maternal, stablize inbred line PW1501 hybridization with the purple Wuta-tsai, first-filial generation with inbred line PW1501 for the male parent that backcrosses continuous 6 generation backcross transformation, select sterility stable, sterile plant rate and sterile degree reach 100% Wuta-tsai Ogura cytoplasm male sterility line.

2. a kind of purple Wuta-tsai male sterile line transfer method as claimed in claim 1, is characterized in that, described Wuta-tsai is stablized inbred line PW1501 seed selection acquisition as follows:

1) select the purple Chinese cabbage PC601 of inheritance stability;

2) select the good Wuta-tsai inbred line of Comprehensive Traits;

3) take purple Chinese cabbage PC601 as maternal, is that male parent is hybridized with the Wuta-tsai selfing, obtains the F1 plant;

4) select purple F1 plant to do maternal, backcross with the Wuta-tsai inbred line, obtain the BC1 seed;

5) do maternally with the purple plant of BC1 seed, with Wuta-tsai inbred line continuous backcross 3-6 time, obtain BCn plant identical with paternal character, the blade purple;

6) purple BCn plant is carried out to selfing and obtain seed BCnS1;

7) BCnS1 is selected to the Wuta-tsai plant that acquisition inheritance stability, leaf look are purple through conventional field test combination;

8) the purple Wuta-tsai plant of acquisition is continued to the Wuta-tsai seed that a large amount of purples of numerous acquisition are expanded in selfing.

3. a kind of purple Wuta-tsai male sterile line transfer method according to claim 2, is characterized in that, described Wuta-tsai inbred line is selected from YW905.

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