CN104160946A - Breeding method for cytoplasmic male sterility line and maintainer line of brassica oleracea - Google Patents
Breeding method for cytoplasmic male sterility line and maintainer line of brassica oleracea Download PDFInfo
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Abstract
The invention discloses a breeding method for cytoplasmic male sterility line and maintainer line of brassica oleracea and application. The method mainly comprises: selecting different types of brassica oleracea materials, inbreeding to obtain a first generation and further to determine a breeding target, directionally selecting a single strain and performing second-generation inbreeding; directionally selecting a single strain from the inbred progeny colony and continuing to perform inbreeding, and directionally selecting a single strain according to the breeding target; also selecting hard-flower type brassica oleracea cytoplasmic male sterility line as a female parent and using the selected single strain as a male parent to perform interspecific hybridization, so as to obtain a hybrid F1 and transfer the CMS gene of the sterile line into brassica oleracea; continuing to perform selfing purification on the inbred progeny, and also screening out a sterile strain closest to the male parent breeding target from the cytoplasmic male sterility line hybrid plant colony and taking the sterile strain as a male parent, taking a single strain closest to the breeding target and screened out from the selfing progeny as a female parent, and continuing to perform paired hybridization; and performing purification of the inbred progeny and transformation of the male sterilty line on each generation at the same time in a paired way, so as to obtain the maintainer line and the cytoplasmic male sterility line after a plurality of generations are obtained through inbreeding. The method is capable of substantially saving labor and saving cost, and shortening breeding time.
Description
Technical field
The present invention relates to a kind of breeding method of preserved egg dish, especially obtain the breeding method of preserved egg dish cytoplasm male sterility line and maintainer, belong to field of plant variety breeding technology.
Background technology
Preserved egg dish (abbreviation of bouquet loose type cauliflower) and common cauliflower comparatively closely bouquet are compared, and bouquet is loose, and bennet is long, green, tastes sweeter, crisp, are difficult for well-donely while stir-frying and eating, and after heating, bennet color becomes greener; Dehydration adds man-hour, take and eats bennet as main, and stalk look dark green, preserved egg dried vegetable unique flavor, and nutritious, liked by consumer.By the analysis to preserved egg dish Main Nutrient Quality, chlorophyll content 12.8 μ gg
-1(FW), carotenoid content 3.3 μ gg
-1(FW), VC content 629mgkg
-1, soluble sugar content 2.82%, with tight type cauliflower ratio, it is high 2.7 times that chlorophyll content is wanted, carotenoid content is high 1.5 times, VC exceeds 40%, it is nearly 10% that soluble sugar exceeds, but aspect moisture, raw fiber, crude protein without significant difference (the Changjiang river vegetables, 2012 (20): 37-39).The mouthfeel that preserved egg dish is good and nutritional characteristic mean that preserved egg dish industry will have huge development prospect, and Substitute For Partial is tightly spent to consumption and the industry demand of type.Preserved egg dish, as far back as Fujian and Taiwan plantation consumption, spreads to China North China, Central China rapidly from area, Zhejiang, Fujian in recent years.China's preserved egg dish major producing area is distributed in the middle and lower reach of Yangtze River at present, Fujian, Zhejiang, high Mountain area, Hubei Province, Lanzhou, Bashang plateau area, and North China, planting area is extensively relatively concentrated again, has basically formed the production pattern in preserved egg dish year-round supply market, and cultivated area has reached 20,000 hm
2above, account for cauliflower and produce 10% of area, the author's prediction, the extensively plantation at home of a few years from now on preserved egg dish, and will become one of main Types of cauliflower consumption, cultivated area will reach 100,000 hm
2above, development potentiality is huge.
Preserved egg dish Heterosis is obvious, and the aspects such as output, quality and resistance all show hybrid vigour.Research shows, utilize cytoplasm male sterility line preparing hybrid kind, have that method is easy, genetic stability, hybrid rate are high, time saving and energy saving, sterility advantage thoroughly, and can effectively control the outflow of merit gene, aspect the crossbreeding of preserved egg dish, have significant application value.And cytoplasmic male sterility (CMS) is a kind of natural phenomena, show as maternal inheritance, pollen abortion and gynoecium normal, with the pollen of fertile plant, give sterile strain gynoecium pollination, can be normally solid.Preserved egg dish is mainly gathered in the crops bouquet trophosome, but not obtains seed, so do not need restorer when hybrid seeding, therefore obtains preserved egg dish cytoplasm male sterility line and maintainer is to utilize the key of male sterile preparing hybrid kind.
Summary of the invention
The technical problem to be solved in the present invention is the defect existing according to prior art, proposes the breeding method of a kind of preserved egg dish cytoplasm male sterility line and maintainer, and the method can significantly save work and economize this, greatly shortens breeding time.
Technical scheme of the present invention realizes by following steps:
(1) collect dissimilar preserved egg dish germ plasm resource, within whole breeding time, observation of plant is learned proterties, disease resistance, yielding ability etc., determines breeding goal, and the directed individual plant of selecting, carries out selfed breeding.
(2) in self progeny colony, select the strain of good, the blue or green stalk preserved egg of disease resistance menu to continue selfing, purify, and according to the directed individual plant of selecting of objective trait.
(3) maternal firmly to spend type Cauliflower cytoplasm male sterility line to do, with directive breeding, individual plant that proterties is good, make male parent, carry out middle intermolecular hybrid, acquisition hybrid F1, thus the sterile gene of male sterile line is transferred on preserved egg dish.
(4), in cytoplasmic male sterilty hybrid plant colony, screening as female parent, continues biparental cross with directive breeding in self progeny, individual plant that proterties is good as male parent with the immediate sterile strain of male parent target; Self progeny continues selfing purification simultaneously.
(5) self progeny's purification and the transformation of male sterile line, in per generation, all carries out in pairs simultaneously, through 6-7 selfing purification and the male sterile line transformation in generation, can obtain cytoplasm male sterility line and maintainer simultaneously; And maintainer has the proterties of good stable, can be used as the parent material of other crossbreed of preparation.
(6) it is maternal take the cytoplasm male sterility line material that institute transformation obtains, and chooses that botany proterties is stable, disease resistance is good, the selfing similar with female parent is male parent, preparing hybrid kind.
In order to reach better effect, preferably:
1, described preserved egg dish inbred line breeding target is: bouquet is pure white, and bennet is dark green, and disease resistance is good, and botany proterties is stable, coordinate force is good, pollen amount is large.
2, the breeding goal of described male sterile line is: sterile plant rate and sterile degree are 100%, and heredity of male sterility is stable, is not subject to the impact of environmental condition; Flower bud development is normal, corolla and petal normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style; Nectary size, quantity and honey amount are physically well developed; Gynoecium structure is normal, can normally set seeds.
3,, in described backcross transformation hybrid population, select the approaching individual plant of the proterties such as plant type, blade profile, bouquet shape, color, disease resistance and male parent to reserve seed for planting.
4, described when selfing purify proceed to 4-5 for time, tree characteristics is no longer separated, then selfing 1-2 generation, can obtain the stable inbred line of isozygotying, and is also maintainer.
5, described 6-7 for backcross transformation cytoplasm male sterility line proterties and maintainer in full accord, its sterile plant rate reaches 100%.
6, described by advance or postpone the mode of cutting bouquet and regulate parents to be consistent at the florescence, and in every mu of seed farm, put into 1-2 case honeybee and carry out supplementary pollination, the isolation distance of seed farm is 1500m at least.
Beneficial effect of the present invention is:
1, through inbreeding of more generation, purify, obtained the inbred line that a large amount of dissimilar botany proterties are stable, coordinate force is good, bouquet proterties is good;
2, through 6-7 for continuous backcross transformation, the male sterile line of acquisition has sterile plant rate and sterile degree is 100%, heredity of male sterility is stable; Flower bud development is normal, corolla and petal normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style; Nectary size, quantity and honey amount are physically well developed; Gynoecium structure is normal, the feature such as can normally set seeds;
The crossbreed of the male sterile line that 3, utilization obtains and the preparation of other inbred line, hybrid vigour is remarkable, botany proterties neat and consistent, sterility is thorough; Saving of labor's abridged edition, has shortened breeding time greatly, has improved breeding efficiency, and can effectively prevent the outflow of specific gene resource, contributes to the protection of independent IP.
Accompanying drawing explanation
Fig. 1 is Breeding Process schematic diagram of the present invention.
Embodiment
Embodiment 1
1) select the preserved egg vegetable kind that is derived from Taiwan, its feature is blue or green stalk, late-maturing; Choose hard colored type Cauliflower cytoplasm male sterility line material B-1 as sterile source.
2) after First selfed progeny, observe characters of progenies separated and change, according to plant type, bouquet shape, blue or green stalk, disease-resistant, high yield, pollen amount is many, compatibility is good breeding goal, directed selection individual plant is reserved seed for planting, and carries out second generation selfing.
3), in selfing 2 generation colony, according to breeding goal, continue the directed individual plant selfing of selecting and purify, and the directed individual plant of selecting carries out third generation selfing.
4) simultaneously and cytoplasm male sterility line material B-1 as maternal, with the selected individual plant of preserved egg dish in 2 generations of selfing, make male parent, hybridize, acquisition hybrid F1, thus the sterile gene of male sterile line is transferred on preserved egg dish.
5) self progeny continues selfing purification; In cytoplasm male sterility line transformation hybrid plant colony, screening is made female parent with the immediate sterile strain of breeding goal, continues biparental cross with the individual plant of directive breeding in self progeny simultaneously.
6) preserved egg dish self progeny's purification and the backcross transformation of cytoplasm male sterility line, in per generation, all carries out in pairs simultaneously, when selfing proceeds to 4-5 for not having proterties obviously separated with backcrossing, selfing and the 1-2 generation that backcrosses again, can obtain the preserved egg dish with objective trait and isozygoty and stablize inbred line and cytoplasm male sterility line, inbred line is also maintainer simultaneously.
7) 6-7 has sterile plant rate for the preserved egg dish cytoplasm male sterility line of backcross transformation and sterile strain degree all reaches 100%, and heredity of male sterility is stable, is not subject to environmental influence; Development of floral organs is normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style, and nectary development is sound; Gynoecium structure is normal, can normally be pollinated and set seeds.
8) take the male sterile based material that obtains is the maternal preserved egg dish hybridization between selfed lines that can educate with other excellence, preparing hybrid kind, during the production of hybrid seeds, by cutting the mode of ball time, regulate parents to be consistent at the florescence, and in every mu of seed farm, put into 1-2 case honeybee and carry out supplementary pollination, the isolation distance of seed farm is 1500m at least.
Sterile plant rate and the sterile degree of the cytoplasm male sterility line that result transformation obtains are 100%, and heredity of male sterility is stable; Flower bud development is normal, corolla and petal normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style; Nectary size, quantity and honey amount are physically well developed; Gynoecium structure is normal, can normally set seeds; Blue or green stalk, high antiviral disease, late-maturing, about 90 days breeding times; The major traits of the maintainer that inbreeding of more generation obtains (inbred line) is: blue or green stalk, and high antiviral is sick, and inheritance stability is late-maturing, about 90 days breeding times.
Embodiment 2
Except step 1) to select and be derived from Japanese commodity kind, blue or green stalk is spent in vain, precocity, high antiviral is sick; Take and firmly spend type Cauliflower cytoplasm male sterility line material B-1 outside sterile source, all the other operate with embodiment 1.
Sterile plant rate and the sterile degree of the cytoplasm male sterility line that result transformation obtains are 100%, and heredity of male sterility is stable; Flower bud development is normal, corolla and petal normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style; Nectary size, quantity and honey amount are physically well developed; Gynoecium structure is normal, can normally set seeds; Blue or green stalk, high antiviral disease, precocity, about 50 days breeding times; The major traits of the maintainer that inbreeding of more generation obtains (inbred line) is: blue or green stalk, and high antiviral is sick, inheritance stability, precocity, about 50 days breeding times.Embodiment 3
Except step 1) select and be derived from Chinese commodity kind, blue or green stalk is spent in vain, in ripe, high antiviral is sick; Take and firmly spend type Cauliflower cytoplasm male sterility line material B-1 outside sterile source, all the other operate with embodiment 1.
Sterile plant rate and the sterile degree of the cytoplasm male sterility line that result transformation obtains are 100%, and heredity of male sterility is stable; Flower bud development is normal, corolla and petal normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style; Nectary size, quantity and honey amount are physically well developed; Gynoecium structure is normal, can normally set seeds; Blue or green stalk, high antiviral are sick, in ripe, about 70 days breeding times; The major traits of the maintainer that inbreeding of more generation obtains (inbred line) is: blue or green stalk, high antiviral is sick, inheritance stability, in ripe, about 70 days breeding times.
Claims (6)
1. a breeding method for preserved egg dish cytoplasm male sterility line and maintainer, comprises the following steps:
(1) collect dissimilar preserved egg dish germ plasm resource, within whole breeding time, observation of plant is learned proterties, disease resistance, yielding ability, determines breeding goal, and the directed individual plant of selecting, carries out selfed breeding;
(2) in self progeny colony, select the strain of good, the blue or green stalk preserved egg of disease resistance menu to continue selfing, purify, and according to the directed individual plant of selecting of objective trait;
(3) maternal firmly to spend type Cauliflower cytoplasm male sterility line to do, with directive breeding, individual plant that proterties is good, make male parent, carry out middle intermolecular hybrid, acquisition hybrid F1, thus the sterile gene of male sterile line is transferred on preserved egg dish;
(4), in cytoplasmic male sterilty hybrid plant colony, screening as female parent, continues biparental cross with directive breeding in self progeny, individual plant that proterties is good as male parent with the immediate sterile strain of male parent target; Self progeny continues selfing purification simultaneously;
(5) self progeny's purification and the transformation of male sterile line, in per generation, all carries out in pairs simultaneously, through 6-7 selfing purification and the male sterile line transformation in generation, can obtain cytoplasm male sterility line and maintainer simultaneously;
(6) it is maternal take the cytoplasm male sterility line material that institute transformation obtains, and chooses that botany proterties is stable, disease resistance is good, the selfing similar with female parent is male parent, and preparing hybrid kind is as filial generation.
2. the breeding method of preserved egg dish cytoplasm male sterility line and maintainer according to claim 1, the breeding goal that it is characterized in that described preserved egg dish inbred line is: bouquet is pure white, bennet is dark green, and disease resistance is good, and botany proterties is stable, coordinate force is good, pollen amount is large.
3. according to the breeding method of preserved egg dish cytoplasm male sterility line and maintainer described in claim 1 or 2, it is characterized in that the breeding goal of male sterile line is: sterile plant rate and sterile degree are 100%, heredity of male sterility is stable, is not subject to the impact of environmental condition; Flower bud development is normal, corolla and petal normal, and it is shrivelled, fine that Anther degeneration becomes, and stamen is significantly shorter than style; Nectary size, quantity and honey amount are physically well developed; Gynoecium structure is normal, can normally set seeds.
4. according to the breeding method of preserved egg dish cytoplasm male sterility line and maintainer described in claim 1 or 2, it is characterized in that in the described filial generation of step (6), select the approaching individual plant of the proterties such as plant type, blade profile, bouquet shape, color, disease resistance and male parent to reserve seed for planting.
5. the breeding method of preserved egg dish cytoplasm male sterility line and maintainer according to claim 1, is characterized in that, the selfing described in step (5) purify proceed to 4-5 for time, tree characteristics is no longer separated, again selfing 1-2 generation, can obtain the stable inbred line of isozygotying, be also maintainer.
6. the breeding method of preserved egg dish cytoplasm male sterility line and maintainer according to claim 5, it is characterized in that, also comprise by advance or postpone the mode of cutting bouquet and regulate parents to be consistent at the florescence, and in every mu of seed farm, put into 1-2 case honeybee and carry out supplementary pollination, the isolation distance of seed farm is 1500m at least.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107347626A (en) * | 2017-07-05 | 2017-11-17 | 张家港市松田创新农业科技有限公司 | A kind of preserved egg dish cultural method |
| CN107581055A (en) * | 2017-08-29 | 2018-01-16 | 云南省农业科学院热区生态农业研究所 | A kind of breeding method of blue or green stalk preserved egg dish |
| CN110301349A (en) * | 2019-08-12 | 2019-10-08 | 浙江省农业科学院 | A kind of selection of green pagoda cauliflower cenospecies |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107347626A (en) * | 2017-07-05 | 2017-11-17 | 张家港市松田创新农业科技有限公司 | A kind of preserved egg dish cultural method |
| CN107581055A (en) * | 2017-08-29 | 2018-01-16 | 云南省农业科学院热区生态农业研究所 | A kind of breeding method of blue or green stalk preserved egg dish |
| CN110301349A (en) * | 2019-08-12 | 2019-10-08 | 浙江省农业科学院 | A kind of selection of green pagoda cauliflower cenospecies |
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