CN102599053A - Method for cultivating Lilium oriental hybrid embryos - Google Patents
Method for cultivating Lilium oriental hybrid embryos Download PDFInfo
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- CN102599053A CN102599053A CN2012100654270A CN201210065427A CN102599053A CN 102599053 A CN102599053 A CN 102599053A CN 2012100654270 A CN2012100654270 A CN 2012100654270A CN 201210065427 A CN201210065427 A CN 201210065427A CN 102599053 A CN102599053 A CN 102599053A
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Abstract
The invention relates to a method for cultivating Lilium oriental hybrid embryos, which is effective in breeding effect and short in breeding period. The method includes: pollinating a Lilium oriental hybrid line; sectioning and peeling small embryos, ovules or ovaries; and performing isolated culture. By cross pollination of the ovaries and young embryos and isolated culture, the embryos can be taken out for culturing before abortion to avoid premature senility of distant hybrid embryos, and accordingly Lilium post-fertilization barrier can be overcome effectively, a great amount of distant hybrid embryos are allowed to continue growing into normal seeds, a breeding procedure is drawn close as soon as possible, and the breeding cycle is shortened. In addition, a great amount of distant hybrids are obtained, Lilium interspecific distant hybridization is achieved, and a batch of novel germplasm is created. The embryo rescue technology and the technology of within-bottle Lilium bulbing are combined, so that culture cycle of Lilium hybrids is shortened effectively, and hybrid survival rate is increased.
Description
Technical field:
The present invention relates to a kind of cultural method of oriental hybrid lily hybrid embryo.
Background technology:
Lily is a kind of important ornamental plants, has higher economic value.Wherein Lilium longiflorum (Longiflorum), Asia lily (Asiatic), oriental hybrid lily (Oriental) and loudspeaker lily (Trumpet) are that commercial most important lily four big hybrids are respectively to have unique merit.Oriental hybrid lily is spent greatly, and pattern is bright-coloured and have strong fragrance, and gray mold (Botrytis) is had very strong resistance, is present domestic most popular lily cut flowers strain.Yet the cutting flower variety of selling in the market all is the kind from external introduction; We also do not have the range of goods of oneself, and it is to carry out the breed improvement and the effective way of the good lily kind of breeding fast that the oriental hybrid lily improved seeds of therefore selecting to have introduced are hybridized.Merit between comprehensive different hybrid system has become an important goal in the lily breeding, but distant hybridization not affinity hindered further developing of lily breeding.The lily distant hybridization exists prefecundation and two kinds of obstacles of after fertilization; The obstacle of after fertilization shows that mainly the growth of after fertilization rataria is imperfect; Hybrid seed hypoplasia after planting all can not germinating growth, and lily hybrid embryo culture technique can effectively solve the after fertilization obstacle.
Summary of the invention:
The present invention is exactly to the problems referred to above, and the cultural method of the oriental hybrid lily hybrid embryo that a kind of breeding effect is good, breeding cycle is short is provided.
In order to realize above-mentioned purpose of the present invention, the present invention adopts following technical scheme, after the system pollination of oriental hybrid lily hybrid, its little embryo, ovule or ovary section is peeled off carry out cultured in vitro.
Concrete incubation step does,
(1) be material with oriental hybrid lily hybrid system, field planting is cultured in the greenhouse blooms;
(2) get the strain of blooming of oriental hybrid lily,, pollinate after 30-40 days, the capsule of its acquisition is taken off carry out sterilization treatment its pollination;
(3) in culture dish, will kill the capsule strip off of bacterium, little embryo, ovule or ovary section are seeded in the medium, inoculative proportion is 1: 20～30 (g/ml
Medium), the cultivation indoor cultivation that is placed on temperature then and is 23～27 ℃ can germinate in 15～45 days, and the germination continued is cultivated after 3～4 months and is cultivated into whole plant.
Little embryo, ovule can germinate through 15 days cultivation;
The ovary slice culture can obviously be seen after the ovule that expands in 30 days, peeled off out the continuation cultivation and can germinate in 15 days.
In culture dish, will kill the capsule one side strip off of bacterium, and have a look at the developmental condition of ovule in the chamber of capsule, the obvious little embryo that can see wire that has just directly strips out and cultivates; The ovule that has maturation is not easy to peel off little embryo, just takes directly to inoculate the ovule cultured method, and what have can only see watery ovule, has transparent kind suitcase, and can't see embryo; The ovule that has also can't see, and just takes the method for ovary slice culture;
In the step (2), oriental hybrid lily hybrid system is pollinated with Asia lily hybrid system.
Sterilizing methods described in the step (2) does, earlier capsule soaked 1-2 second in 75% alcohol, clips with tweezers then and is put into the burning of raising above the alcolhol burner flame, carries out surface disinfection.
In the step (3) little embryo being carried out the used medium of cultured in vitro is 1/2MS+BA0.05～0.2mg/l+NAA 0.05～0.2mg/l+ sucrose 50000mg/l.
In the step (3) ovule being carried out the used medium of cultured in vitro is 1/2MS+BA0.05～0.2mg/l+ sucrose 40000mg/l.
In the step (3) the used medium of cultured in vitro being carried out in the ovary section is MS+NAA0.1～0.3mg/l+ sucrose 30000mg/l.
Beneficial effect of the present invention:
The present invention provides the nutrition of hybrid rataria through the medium of manual work preparation, makes it continued growth and grows until Cheng Miao.Hybridization ovary, ovule and hybridization rataria cultured in vitro are the effective ways that overcome the after fertilization obstacle at present, and be wherein maximum with rataria cultured in vitro chance of success.The rataria cultured in vitro is used for saving at abortion of the middle and later periods of embryonic development or jejune species hybrid embryo widely.
Cultured in vitro through hybridization pollination ovary and rataria can be taken out cultivation with it before the embryo abortion, avoid the early ageing of distant hybrid embryo, effectively solved lily after fertilization obstacle; Make a large amount of distant hybridization embryos continue to develop into normal seed; Get into the seed selection program as early as possible, shortened breeding cycle, obtained a large amount of distant hybrid progenies simultaneously; Realized the lilium interspecific hybridization; Created a collection of new germ plasm, embryo rescue techniques combines with the interior balling technology of lily bottle, has effectively shortened hybrid lily offspring's cultivation period and has heightened the hybrid survival rate.
Embodiment:
Embodiment 1
With the east hybrid lily is that kind ' Suo Bang ' is a female parent; With the Asia hybrid lily is that kind ' Reno ' is a male parent; The capsule that pollination was obtained in back 36 days downcuts, and on superclean bench, it is carried out surface disinfection and handles, and promptly earlier capsule is soaked 1-2 second in 75% alcohol; Clip with tweezers then and be put into the alcolhol burner flame top burning of raising, carry out surface disinfection.Then in culture dish, will kill the capsule one side strip off of bacterium; Little embryo is directly stripped out; Be inoculated into and consist of in the 1/2MS+BA 0.1mg/l+NAA 0.1mg/l+ sucrose 50000mg/l medium, inoculative proportion is 1: 20g/ml, and the cultivation indoor cultivation that then is placed on temperature and is 23 ℃ successfully germinateed after 15 days; Continue to cultivate after 3 months and cultivate into whole plant, cultivate into lily new varieties ' virtue is fragrant '.Use above-mentioned cultural method hybrid survival rate and reach 91%.
' virtue is fragrant ' basic proterties: brightly yellowish look, immaculate, flower footpath 15cm, the long 8cm of petal, the wide 4.5cm of petal, sepal warp degree is little, and column cap is yellow, British plain spirits.Plant height 56cm, 10 weeks of vegetative period; Overall assessment is good.
Embodiment 2
With the east hybrid lily is that kind ' expression ' is a female parent; With the Asia hybrid lily is that kind ' Wal rice ' is a male parent; The capsule that pollination was obtained in back 40 days downcuts, and on superclean bench, it is carried out surface disinfection and handles, and promptly earlier capsule is soaked 1-2 second in 75% alcohol; Clip with tweezers then and be put into the alcolhol burner flame top burning of raising, carry out surface disinfection.Then in culture dish, will kill the capsule one side strip off of bacterium; Little embryo is directly stripped out; Be inoculated into to consist of in the 1/2MS+BA 0.2mg/l+NAA 0.05mg/l+ sucrose 50000mg/l medium and cultivate, inoculative proportion is 1: 30g/ml, and the cultivation indoor cultivation that then is placed on temperature and is 27 ℃ successfully germinateed after 15 days; Continue to cultivate after 4 months and cultivate into whole plant, cultivate into lily new varieties ' red red '.Use above-mentioned cultural method hybrid survival rate and reach 94%.
' red red ' basic proterties: petal is yellow; The petal base portion is red, and the edge is yellow, and petal separates, and purple dot is arranged, flower footpath 14cm, and the long 8cm of petal, the wide 3.7cm of petal, sepal warp degree is little, and column cap is yellow, British plain spirits.Potted flower, plant height 50cm, 12 weeks of vegetative period; Overall assessment is good.
The parent who relates to ' Reno ', ' Wal rice ', ' Suo Bang ', ' expression ' all buy the company from Zhejiang Hongyue Flowers Co., Ltd.
Claims (7)
1. the cultural method of an oriental hybrid lily hybrid embryo is characterized in that, after the pollination of oriental hybrid lily hybrid system, its little embryo, ovule or ovary section is peeled off, and carries out cultured in vitro.
2. the cultural method of oriental hybrid lily hybrid embryo according to claim 1 is characterized in that, concrete incubation step does,
(1) be material with oriental hybrid lily hybrid system, field planting is cultured in the greenhouse blooms;
(2) get the strain of blooming of oriental hybrid lily,, pollinate after 30-40 days, the capsule of its acquisition is taken off carry out sterilization treatment its pollination;
(3) in culture dish, will kill the capsule strip off of bacterium; Little embryo, ovule or ovary section are inoculated in the medium; Inoculative proportion is 1: 20～30g/ml; The cultivation indoor cultivation that is placed on temperature then and is 23～27 ℃ can germinate in 15～45 days, and the germination continued is cultivated after 3～4 months and cultivated into whole plant.
3. the cultural method of oriental hybrid lily hybrid embryo according to claim 2 is characterized in that, in the step (2), with Asia lily hybrid system oriental hybrid lily hybrid system is pollinated.
4. the cultural method of oriental hybrid lily hybrid embryo according to claim 2; It is characterized in that the sterilizing methods described in the step (2) is earlier capsule to be soaked 1-2 second in 75% alcohol; Clip with tweezers then and be put into the alcolhol burner flame top burning of raising, carry out surface disinfection.
5. the cultural method of oriental hybrid lily hybrid embryo according to claim 2 is characterized in that, in the step (3) little embryo being carried out the used medium of cultured in vitro is 1/2MS+BA0.05～0.2mg/l+NAA 0.05～0.2mg/l+ sucrose 50000mg/l.
6. the cultural method of oriental hybrid lily hybrid embryo according to claim 2 is characterized in that, in the step (3) ovule being carried out the used medium of cultured in vitro is 1/2MS+BA0.05～0.2mg/l+ sucrose 40000mg/l.
7. the cultural method of oriental hybrid lily hybrid embryo according to claim 2 is characterized in that, in the step (3) the used medium of cultured in vitro being carried out in the ovary section is MS+NAA0.1～0.3mg/l+ sucrose 30000mg/l.
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| CN2012100654270A CN102599053A (en) | 2012-03-14 | 2012-03-14 | Method for cultivating Lilium oriental hybrid embryos |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102907313A (en) * | 2012-11-03 | 2013-02-06 | 云南省农业科学院花卉研究所 | Method of constructing clone groups of lily hybridization generation |
| CN103782910A (en) * | 2014-02-13 | 2014-05-14 | 中国农业科学院蔬菜花卉研究所 | Isolated culture method of unfertilized ovary of lilium |
| CN105123528A (en) * | 2015-09-18 | 2015-12-09 | 北京林业大学 | Method for embryo rescue at different developmental stages of early phase of lilium formolongi distant hybridization |
| CN112790104A (en) * | 2020-12-30 | 2021-05-14 | 云南农业大学 | Method for identifying affinity of lily distant hybridization |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1994067A (en) * | 2006-11-16 | 2007-07-11 | 云南省农业科学院花卉研究所 | Method for obtaining lily distant hybrid by using rescue isolated culture of buds |
| CN101411304A (en) * | 2008-05-19 | 2009-04-22 | 贵州省园艺研究所 | Method for cultivating plant strain of distant hybrid lily |
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2012
- 2012-03-14 CN CN2012100654270A patent/CN102599053A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1994067A (en) * | 2006-11-16 | 2007-07-11 | 云南省农业科学院花卉研究所 | Method for obtaining lily distant hybrid by using rescue isolated culture of buds |
| CN101411304A (en) * | 2008-05-19 | 2009-04-22 | 贵州省园艺研究所 | Method for cultivating plant strain of distant hybrid lily |
Non-Patent Citations (2)
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| 刑大洲: "百合胚拯救育种技术及引进百合新品种筛选研究", 《北京林业大学》 * |
| 郑思乡等: "黄花百合与东方百合‘如意’远缘杂交的研究", 《西部林业科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102907313A (en) * | 2012-11-03 | 2013-02-06 | 云南省农业科学院花卉研究所 | Method of constructing clone groups of lily hybridization generation |
| CN103782910A (en) * | 2014-02-13 | 2014-05-14 | 中国农业科学院蔬菜花卉研究所 | Isolated culture method of unfertilized ovary of lilium |
| CN103782910B (en) * | 2014-02-13 | 2016-01-06 | 中国农业科学院蔬菜花卉研究所 | The method of lily Unfertilized Ovaries cultured in vitro |
| CN105123528A (en) * | 2015-09-18 | 2015-12-09 | 北京林业大学 | Method for embryo rescue at different developmental stages of early phase of lilium formolongi distant hybridization |
| CN105123528B (en) * | 2015-09-18 | 2018-03-30 | 北京林业大学 | New PE curriculum distant hybridization early stage different developmental phases embryo rescue method |
| CN112790104A (en) * | 2020-12-30 | 2021-05-14 | 云南农业大学 | Method for identifying affinity of lily distant hybridization |
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Application publication date: 20120725 |