CN105104170A - Cultivating method for stevia rebaudiana triploid hybrid seeds - Google Patents
Cultivating method for stevia rebaudiana triploid hybrid seeds Download PDFInfo
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- CN105104170A CN105104170A CN201510517754.9A CN201510517754A CN105104170A CN 105104170 A CN105104170 A CN 105104170A CN 201510517754 A CN201510517754 A CN 201510517754A CN 105104170 A CN105104170 A CN 105104170A
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Abstract
The invention discloses a cultivating method for stevia rebaudiana triploid hybrid seeds. Chromosome doubling is conducted on diploid materials of stevia rebaudiana, then tissue culture propagation is conducted on tetraploid and diploid stevia rebaudiana single plant materials which are to be paired and hybridized; a plurality of seed production parent strains of tetraploid and diploid stevia rebaudiana are obtained respectively, the seed production parent strains of tetraploid and diploid stevia rebaudiana are consistent in genotype and reproduction character, the two kinds of seed production parent strains are planted in the same field alternately in rows to conduct hybrid seed production, and due to the fact that stevia rebaudiana has the character of selfing barrenness, the stevia rebaudiana triploid hybrid seeds can be obtained after natural hybridization. The blade yield of stevia rebaudiana can be obviously increased through the measure according to the giant property of plant triploid organs.
Description
Technical field
a breeding method for stevia triploid hydrid kind, relates to stevia triploid Breeding technology, is that a kind of dliploid stevia and tetraploid stevia carry out hybridizing the method for cultivating triploid stevia crossbreed in particular.
Background technology
in crops, many crops have all been bred as triploid high yield and high quality new varieties, bring breakthrough raising to the yield and quality of crop.Its specific practice, for watermelon, is bred as seedling by the watermelon seed of zygoid and the tetraploid watermelon seed that isozygotys, is alternately planted on same soil, and allow them carry out the hybridization of orientation, namely the seed obtained is triploid watermelon seed.It is all adopt conventional kind or even the farm variety of various places that current stevia is produced, and yields poorly and unstable, and quality is uneven.Triploid plant has the huge feature of reproductive organs, if can be used in stevia, the blade of stevia also can be made huge, thus obtains higher output.But due to characteristic and the botanical character of the hereditary reproduction of stevia self, the triploid breeding of stevia can not apply mechanically the method for other crop simply.Stevia is a kind of plant of strict cross pollination, has selfing sterility, but the pollen between different plants can be set seeds in phase mutual cross.If dliploid and tetraploid Stevia seed seed are implanted in same piece of land, not only dliploid and tetraploid can phase mutual crosses, also can pollinate hybridization mutually in Diploid and Tetraploid colony inside, so the seed produced will be disorderly and unsystematic, existing triploid crossbreed, also there are tetraploid and diplontic seed, make colony there is no uniformity, agricultural production standardisation requirements can not be reached.Because flower amount is huge, fruit is very little, and the seed that every flower is tied is few, obviously cannot produce Stevia seed in a large number with manually carrying out hybridizing.How the dliploid of stevia and tetraploid being hybridized, produce consistent stevia triploid hydrid kind, is a difficult problem urgently to be resolved hurrily.
Summary of the invention
the object of the invention is to the above-mentioned difficult problem for stevia triploid Breeding exists, propose a kind of can hybridization by the Diploid and Tetraploid of stevia, produce the method for consistent stevia triploid hydrid kind.
the present invention is achieved in that
a breeding method for stevia triploid hydrid kind, it comprises the steps:
(1) mutagenesis stevia tetraploid: double to become tetraploid with the dliploid material mutagenesis of colchicine by part stevia;
(2) stevia production of hybrid seeds parent strain is cultivated: under condition of tissue culture, expansion is carried out to the stevia individual plant material of Diploid and Tetraploid numerous, breed out stevia production of hybrid seeds parent strain all consistent with reproductive characteristic from the genotype of single plant in a large number respectively;
(3) hybrid seeding: be planted in same Tanaka by alternate by row for the body stevia production of hybrid seeds parent strain of tetraploid stevia production of hybrid seeds parent's strain and two times, utilize insect to carry out mutual hybridization pollination, the seed of results is triploid Stevia seed.
the described tetraploid method of mutagenesis stevia is: on superclean bench, take out test-tube plantlet removing blade, be cut into about 1cm long containing side leaf stem section, be placed horizontally at regeneration culture medium on the surface, flicking is fixed, every bottle graft kind 5 stem sections, each process inoculation 10 bottles, finally be placed in culturing room to carry out taking out after 2d is cultivated in cultivation, superclean bench drips 0.025% colchicine (filtration sterilization), guarantee that colchicine infiltrates each stem section, colchicine is disposed and puts back to culturing room, continue to cultivate (about 30d) when bud seedling grows to 2.0cm height, bud seedling is cut to be inoculated in root media by superclean bench and cultivates 20d, seize and double successful tetraploid plant.
described regeneration culture medium is leaf regeneration seedling medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ agar powder 5.0g/L+ sucrose 30g/L; Culture of rootage is root media is MS+IBA0.5mg/L+ agar powder 5.0g/L+ sucrose 20g/L.
described stevia expands numerous adopted medium by the stage of cultivating, and is divided into regeneration culture medium: MS+6-BA1.0mg/L+NAA0.2mg/L+ agar powder 5.0g/L+ sucrose 30g/L and root media: MS+IBA0.5mg/L+ agar powder 5.0g/L+ sucrose 20g/L.
beneficial effect of the present invention:
the present invention proposes under conditions of tissue culture, with colchicine, the induction of part stevia material is doubled the body that quadruples, again respectively by the method for group training, the production of hybrid seeds parent strain of the single Diploid and Tetraploid of numerous successful genotype is expanded in amplification in enormous quantities by single plant.Its genotype separately in colony between individuality is just the same, is all that a stevia individual plant " is attended to anything else ", reproductive characteristic can be considered an individual plant.Because stevia has the acarpous characteristic of selfing, not only individual selfing can not be solid for such production of hybrid seeds parent strain, can not mutually pollinate solid between the plant of its colony inside separately.The Diploid and Tetraploid stevia seed cultivated like this is implanted in together, when blooming, only has between the plant of Diploid and Tetraploid and can phase mutual cross set seeds, so just obtain genotype triploid Stevia seed all consistent with reproductive characteristic.This method effectively prevent the hybridization of Stevia seed seedling, and the plant phase mutual cross in the parental population of existence, causes the problem that seed seriously mixes.Because triploid plant has the rule of the huge property of organ, thus the output of stevia is significantly improved.
Embodiment
cultivate the step of stevia triploid hydrid kind:
(1) mutagenesis stevia tetraploid: double to become tetraploid with the dliploid material mutagenesis of colchicine by part stevia;
(2) stevia production of hybrid seeds parent strain is cultivated: under condition of tissue culture, expansion is carried out to the stevia individual plant material of Diploid and Tetraploid numerous, breed out stevia production of hybrid seeds parent strain all consistent with reproductive characteristic from the genotype of single plant in a large number respectively;
(3) hybrid seeding: be planted in same Tanaka by alternate by row for the body stevia production of hybrid seeds parent strain of tetraploid stevia production of hybrid seeds parent's strain and two times, utilize insect to carry out mutual hybridization pollination, the seed of results is triploid Stevia seed.
the tetraploid method of mutagenesis stevia is: on superclean bench, take out test-tube plantlet removing blade, be cut into about 1cm long containing side leaf stem section, be placed horizontally at regeneration culture medium on the surface, flicking is fixed, every bottle graft kind 5 stem sections, each process inoculation 10 bottles, finally be placed in culturing room to carry out taking out after 2d is cultivated in cultivation, superclean bench drips 0.025% colchicine (filtration sterilization), guarantee that colchicine infiltrates each stem section, colchicine is disposed and puts back to culturing room, continue to cultivate (about 30d) when bud seedling grows to 2.0cm height, bud seedling is cut to be inoculated in root media by superclean bench and cultivates 20d, seize and double successful tetraploid plant.
regeneration culture medium is leaf regeneration seedling medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ agar powder 5.0g/L+ sucrose 30g/L; Culture of rootage is root media is MS+IBA0.5mg/L+ agar powder 5.0g/L+ sucrose 20g/L.
stevia expands numerous adopted medium by the stage of cultivating, and is divided into regeneration culture medium: MS+6-BA1.0mg/L+NAA0.2mg/L+ agar powder 5.0g/L+ sucrose 30g/L and root media: MS+IBA0.5mg/L+ agar powder 5.0g/L+ sucrose 20g/L.
Claims (4)
1. a breeding method for stevia triploid hydrid kind, is characterized in that, it comprises the steps:
(1) mutagenesis stevia tetraploid: double to become tetraploid with the dliploid material mutagenesis of colchicine by part stevia;
(2) stevia production of hybrid seeds parent strain is cultivated: under condition of tissue culture, expansion is carried out to the stevia individual plant material of Diploid and Tetraploid numerous, breed out stevia production of hybrid seeds parent strain all consistent with reproductive characteristic from the genotype of single plant in a large number respectively;
(3) hybrid seeding: be planted in same Tanaka by alternate by row for the body stevia production of hybrid seeds parent strain of tetraploid stevia production of hybrid seeds parent's strain and two times, utilize insect to carry out mutual hybridization pollination, the seed of results is triploid Stevia seed.
2. the breeding method of a kind of stevia triploid hydrid kind according to claim 1, it is characterized in that, the described tetraploid method of mutagenesis stevia is: on superclean bench, take out test-tube plantlet removing blade, be cut into about 1cm long containing side leaf stem section, be placed horizontally at regeneration culture medium on the surface, flicking is fixed, every bottle graft kind 5 stem sections, each process inoculation 10 bottles, finally be placed in culturing room to carry out taking out after 2d is cultivated in cultivation, superclean bench drips 0.025% colchicine (filtration sterilization), guarantee that colchicine infiltrates each stem section, colchicine is disposed and puts back to culturing room, continue to cultivate (about 30d) when bud seedling grows to 2.0cm height, bud seedling is cut to be inoculated in root media by superclean bench and cultivates 20d, seize and double successful tetraploid plant.
3. the breeding method of a kind of stevia triploid hydrid kind according to claim 2, is characterized in that, described regeneration culture medium is leaf regeneration seedling medium is MS+6-BA1.0mg/L+NAA0.2mg/L+ agar powder 5.0g/L+ sucrose 30g/L; Culture of rootage is root media is MS+IBA0.5mg/L+ agar powder 5.0g/L+ sucrose 20g/L.
4. the breeding method of a kind of stevia triploid hydrid kind according to claim 2, it is characterized in that, described stevia expands numerous adopted medium by the stage of cultivating, and is divided into regeneration culture medium: MS+6-BA1.0mg/L+NAA0.2mg/L+ agar powder 5.0g/L+ sucrose 30g/L and root media: MS+IBA0.5mg/L+ agar powder 5.0g/L+ sucrose 20g/L.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02242622A (en) * | 1989-03-16 | 1990-09-27 | Sanyo Kokusaku Pulp Co Ltd | Stevia triploid and method for proliferation thereof |
| CN102893781A (en) * | 2012-09-24 | 2013-01-30 | 林平 | Stevia rebaudiana single-joint half-stem cottage method |
| CN102893860A (en) * | 2011-10-19 | 2013-01-30 | 崔广荣 | Stevia rebaudiana bertoni polyploid in vitro induction technology |
-
2015
- 2015-08-23 CN CN201510517754.9A patent/CN105104170A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02242622A (en) * | 1989-03-16 | 1990-09-27 | Sanyo Kokusaku Pulp Co Ltd | Stevia triploid and method for proliferation thereof |
| CN102893860A (en) * | 2011-10-19 | 2013-01-30 | 崔广荣 | Stevia rebaudiana bertoni polyploid in vitro induction technology |
| CN102893781A (en) * | 2012-09-24 | 2013-01-30 | 林平 | Stevia rebaudiana single-joint half-stem cottage method |
Non-Patent Citations (1)
| Title |
|---|
| 彭程等: "低浓度秋水仙素离体诱导甜叶菊多倍体技术体系的建立", 《热带作物学报》 * |
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