CN1255023C - Quick breeding technolgy for Renanthera imschootiana Rolfe - Google Patents
Quick breeding technolgy for Renanthera imschootiana Rolfe Download PDFInfo
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- CN1255023C CN1255023C CN 200410050909 CN200410050909A CN1255023C CN 1255023 C CN1255023 C CN 1255023C CN 200410050909 CN200410050909 CN 200410050909 CN 200410050909 A CN200410050909 A CN 200410050909A CN 1255023 C CN1255023 C CN 1255023C
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Abstract
The present invention relates to a sterile sowing and tissue culture technique for Renanthera imschootiana Rolfe. Renanthera imschootiana Rolfe plants have fifteen kinds and have very high ornamental value. International commercial seedlings for Renanthera imschootiana Rolfe are produced by manual sowing and tissue culture propagation at present, and the seedlings of Renanthera imschootiana Rolfe can not be produced in our nation. The propagation of Renanthera imschootiana Rolfe is often carried out by a plant division method, but the reproduction speed is low. Because the growth of the embryos of the seeds of Renanthera imschootiana Rolfe is incomplete, the seeds of Renanthera imschootiana Rolfe are extremely difficult to germinate in a natural state. Compared with the prior art, the present invention has the advantages of high seed germination rate, less cultivation time of test-tube seedlings, robust test-tube seedlings, strong operability and high application value. The pot flowers and cut flowers for Renanthera imschootiana Rolfe have large market requirements, and the present invention has good application foreground.
Description
Technical field
The present invention relates to the aseptic seeding and the tissue culture technique of Herba Renantherae coccineae.
Background technology
The Herba Renantherae coccineae platymiscium has 15 kinds, and China originates in 2 kinds, all has very high ornamental value, its Hybrid is various, can carry out intergeneric cross with a plurality of genus, with Moth orchid, all ages orchid etc. hybridizes the cut-flower and the potted plant kind of many top grades, is rare kind emerging in world's orchid industry.Present international Herba Renantherae coccineae commercial seedling all will be produced by by artificial seeding and tissue culture propagation, the domestic seedling production of still not having Herba Renantherae coccineae, and the market demand of the potted flower of Herba Renantherae coccineae and cut-flower is big, present technique has good application prospects.
Offshoot is adopted in the breeding of Herba Renantherae coccineae more, and reproduction speed is slow; And the Herba Renantherae coccineae seed is incomplete owing to embryonic development, extremely difficult sprouting the under the nature, but on suitable synthetic medium, can carry out.The application adopts the method for aseptic seeding can obtain a large amount of test-tube plantlets, and the protocorm that utilizes aseptic seeding to obtain can be bred in a large number.
At present, the domestic report that does not have Herba Renantherae coccineae aseptic seeding and Study on tissue culture and large-scale production and patent application.Patent of the present invention provides the method and the employed medium of the aseptic seeding of Herba Renantherae coccineae and tissue culture propagating different with international existing report, the seed germination rate height, short, the robust growth of test-tube plantlet incubation time, have workable, the advantage that using value is high.
Summary of the invention
The purpose of this invention is to provide the quick propagating technology prescription and the production procedure of Herba Renantherae coccineae, this technology is fit to suitability for industrialized production very much, and the prescription effect is preferable, germination rate, and rate of increase height, plant strain growth is vigorous.The propagation method of the Herba Renantherae coccineae seedling that provides and employed medium, cost is low, and is workable.
The characteristics of this technology are:
1, technology of the present invention is Herba Renantherae coccineae (Renanthera coccinea), Yunnan Herba Renantherae coccineae (Renantheraimschootiana), leopard's spots Herba Renantherae coccineae (Renanthera monachica) and orange Herba Renantherae coccineae (Renanthera citrus) for the examination material.
2, the technology of the present invention content is aseptic seeding and grows seedlings, and the optimization culture medium prescription is provided, and seed germination rate is up to 85%, and seedling early growth is rapid, robust plant.
3, during test-tube seedling transplanting, be natural daylight lower refining seedling 7-14 days, transplanting survival rate is up to 95-100%.
4, Herba Renantherae coccineae is rare ornamental plants, and breeding is main adopts division propagation, but reproduction speed is slow, and the Herba Renantherae coccineae seed does not have endosperm because embryonic development is incomplete, extremely difficult sprouting the under the nature.This patent adopts the method for aseptic seeding can obtain a large amount of test-tube plantlets in a short time, and the protocorm that utilizes aseptic seeding to produce can be bred.
5, the technology of the present invention can successfully be carried out aseptic seeding and the tissue-culturing quick-propagation of Herba Renantherae coccineae original seed and crossbreed thereof, has simple effective, the less investment of prescription, the characteristics that output is high.Implementing this patent only need have simple Plant Tissue Breeding equipment to carry out.
The quick propagating technology of Herba Renantherae coccineae of the present invention may further comprise the steps:
1. material: Herba Renantherae coccineae (Renanthera coccinea), Yunnan Herba Renantherae coccineae (Renantheraimschootiana), leopard's spots Herba Renantherae coccineae (Renanthera monachica) and orange Herba Renantherae coccineae (Renanthera citrus) maternal plant of choosing robust growth when blooming carry out artificial pollination, produce selfing and hybridization fruit, be used for sowing as explant when pollination back fruit was mature on the whole about 120 days.
2. aseptic seeding: with 75% alcohol-pickled 30 seconds being placed in 0.1% the mercuric chloride solution sterilization 20 minutes, cut fruit after the rinsed with sterile water 4～5 times, with transfer needle the white powder seed is inoculated into seed germination medium last 30 day of left and right sides embryo germination, form complete plantlet about 60 days, in time plantlet is changed over to the strong seedling culture base.
3. enrichment culture: for obtaining more seedling, the protocorm that aseptic seeding is produced is cultivated and can form protocorms on proliferated culture medium, and the rate of increase is more than 5, and 30 days is a subculture cycle.
4. strong plantlets and rootage is cultivated: protocorm is at the long-time seedling that can form unrooted in about 60 days of cultivating on seed culture or the proliferated culture medium, in time plant is cut into individual plant, be inoculated on the strong plantlets and rootage medium and cultivate, 4 weeks can form whole plant, and the plant in 6-8 week can carry out acclimatization and transplants.
5. test-tube seedling transplanting: when test-tube plantlet strong seedling culture after 60 days (plant has 3-4 sheet leaf approximately), transferred to the natural daylight lower refining seedling 7-14 days, then it is taken out from vial, clean the medium of root, plant in 1.5 cun basins with thick colored sphagna, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach 95-100%.
The composition of the used medium of this method is:
Seed germination medium: coconut milk+activated carbon 0.5-2 grams per liter of the precious medium 1 of compound flower (spend precious No. one 2 gram, spend precious No. 21 grams)+ND medium organic component+6-benzyl purine 0.5-2 mg/litre+methyl 0.2-1 mg/litre+10-20%.
Proliferated culture medium: coconut milk+activated carbon 0.5-2 grams per liter of the precious medium 1+ND of compound flower medium organic component+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+10-20%.
Strong plantlets and rootage medium: the precious medium 2 of compound flower (spend precious No. one 1 gram, spend precious No. 22 grams)+banana homogenate 100 grams per liters+6-benzyl purine 0.1-0.5 mg/litre+methyl 0.5-2 mg/litre+activated carbon 0.5-2 grams per liter.
Above medium all contains sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%.Cultivation temperature (28 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
The quick propagating technology of Herba Renantherae coccineae of the present invention compared with prior art has the germination rate height, and it is fast to emerge, rate of increase height, and seedling is strong, advantages such as seedling quality better, well-grown.The present invention simultaneously is broadly representative for the examination material, can be as other kind of flame Cymbidium, belong in or with the method for other aseptic seeding of bigenering and tissue culture.
Embodiment:
Example 1:
1, obtains seed material: choose the Herba Renantherae coccineae (renanthera coccinia) of greenhouse culture, the healthy and strong plant that leopard's spots Herba Renantherae coccineae (renantheramonachita) is bloomed, with the Herba Renantherae coccineae is maternal, the leopard's spots Herba Renantherae coccineae is that male parent is hybridized, the visible column cap closure of pollinating after 2 days, ovary germinates, pollinate after 130 days, visible fruit changes yellow, takes off fruit and uses for sowing.
2, the sterilization method of fruit: the fruit that will take off from maternal plant is with 75% alcohol-pickled 30 seconds being placed on 0.1% the mercuric chloride solution sterilization 20 minutes, cuts fruit behind the aseptic water washing 4～5 times.
3, seed germination: the white powder embryo is inoculated into germination medium: coconut milk+activated carbon 0.5 grams per liter of the precious medium 1 of compound flower (spend precious No. one 2 gram, spend precious No. 21 grams)+ND medium organic component+6-benzyl purine 0.5 mg/litre+methyl 0.5 mg/litre+10% with transfer needle.30 days left and right sides embryo germinations, germination rate about 85%.
4, enrichment culture: the PLB that seed germination is produced is transferred to proliferated culture medium: coconut milk+6-benzyl purine 2 mg/litre+methyl 0.2 mg/litre+activated carbon 0.5 grams per liter of the precious medium 1+ND of compound flower medium organic component+10%.Add active carbon and help preventing brownization, the rate of increase is more than 6, and subculture cycle is 30 days.
5, strong plantlets and rootage is cultivated: the protocorms on proliferated culture medium or seed germination medium is cultivated the seedling that can form unrooted about 60 days, these seedlings is cut into individual plant is inoculated into the strong seedling culture base: the precious medium 2 of compound flower (spend precious No. one 1 gram, spend precious No. 22 grams)+banana homogenate 100 grams per liters+6-benzyl purine 0.1 mg/litre+methyl 1 mg/litre+activated carbon 1 grams per liter.Cultivating for 2 weeks is that visible root grows, and can form the transplanted seedling that has 3-5 sheet leaf after cultivating for 6 weeks.
6, test-tube plantlet hardening and transplanting: the whole bottle of the test-tube plantlet that can transplant is positioned over 1 week of hardening in the greenhouse of 70% shading, take out seedling, medium is cleaned, dry bacterium liquid after 500 times of carbendazim aqueous solution soaking, plant in 2.5 cun little basins with superfine sphagna, keep suitably ventilating and enough humidity, the survival rate of transplanting can reach 100%.
Above medium all contains sucrose 20 grams per liters, pH 5.2-5.4, agar 0.7%.Cultivation temperature (28 ± 2) ℃, illuminance 1500～2000lx, illumination 12 hours/day.
Example 2:
The Different Individual of choosing the healthy and strong plant of Yunnan Herba Renantherae coccineae (renanthera imschootiana) of greenhouse culture is a parental source, carried out selfing in back 2 days in blooming, the visible column cap closure of pollinating after 2 days, ovary germinates, pollinate after 120 days, as seen fruit changes yellow, takes off fruit and uses for sowing.Other basic operation methods are with embodiment 1 unanimity.But its seed germination medium is: coconut milk+activated carbon 0.5 grams per liter of the precious medium 1 of compound flower (spend precious No. one 2 gram, spend precious No. 21 grams)+ND medium organic component+6-benzyl purine 0.5 mg/litre+methyl 0.5 mg/litre+20%, its germination rate is the highest by about 60%.The strong plantlets and rootage medium is the precious medium 2 of compound flower (spend precious No. one 1 gram, spend precious No. 22 grams)+banana homogenate 100 grams per liters+6-benzyl purine 0.5 mg/litre+methyl 1 mg/litre+activated carbon 2 grams per liters.The survival rate of test-tube seedling transplanting can reach 95%.
Claims (2)
1, a kind of method for quickly breeding of Herba Renantherae coccineae is characterized in that this method may further comprise the steps:
1), material: the Herba Renantherae coccineae maternal plant of choosing robust growth when blooming carries out artificial pollination, produces selfing and hybridization fruit, when pollination back fruit 120 days is ripe, is used for sowing as explant;
2), aseptic seeding: explant is with 75% alcohol-pickled 30 seconds being placed in 0.1% the mercuric chloride solution sterilization 20 minutes, cut fruit after the rinsed with sterile water 4～5 times, with transfer needle the white powder seed is inoculated into seed germination medium last 30 day of embryo germination, 28 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day; Formed complete plantlet in 60 days, in time plantlet is changed over to the strong seedling culture base, the seed germination medium is: coconut milk+activated carbon 0.5-2 grams per liter of the precious medium 1+ND of compound flower medium organic component+6-benzyl purine 0.5-2 mg/litre+methyl 0.2-1 mg/litre+10-20%; Contain sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%; The precious medium 1 of described compound flower is: spend precious No. one 2 gram, spend precious No. 21 grams;
3), enrichment culture: for obtaining more seedling, the protocorm cultivation that aseptic seeding is produced can form protocorms, 28 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day on proliferated culture medium; 30 days is a subculture cycle, proliferated culture medium is: coconut milk+activated carbon 0.5-2 grams per liter of the precious medium 1+ND of compound flower medium organic component+6-benzyl purine 1-5 mg/litre+methyl 0.2-0.5 mg/litre+10-20%, contain sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%; The precious medium 1 of compound flower is for spending precious No. one 2 gram, spending precious No. 21 grams;
4), strong plantlets and rootage is cultivated: protocorm is the long-time seedling that can form unrooted in 60 days of cultivating on seed culture or proliferated culture medium, in time plant is cut into individual plant, is inoculated on the strong plantlets and rootage medium and cultivates, 28 ± 2 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day; 4 weeks can form whole plant, the plant in 6-8 week can carry out acclimatization and transplants, the strong plantlets and rootage medium is: the precious medium 2+ of compound flower banana homogenate 100 grams per liters+6-benzyl purine 0.1-0.5 mg/litre+methyl 0.5-2 mg/litre+activated carbon 0.5-2 grams per liter, contain sucrose 20 grams per liters, pH5.2-5.4, agar 0.7%; The precious medium 2 of compound flower is for spending precious No. one 1 gram, spending precious No. 22 grams;
5), test-tube seedling transplanting: when test-tube plantlet strong seedling culture after 60 days, plant has 3-4 sheet leaf approximately, transfers to the natural daylight lower refining seedling 7-14 days, then it is taken out from vial, clean the medium of root, plant in 1.5 cun basins, keep suitably ventilating and enough humidity with thick flower sphagna.
2, according to the method for quickly breeding of the Herba Renantherae coccineae in the claim 1, it is characterized in that 1) described in the Herba Renantherae coccineae maternal plant be Yunnan Herba Renantherae coccineae, leopard's spots Herba Renantherae coccineae or orange Herba Renantherae coccineae.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410050909 CN1255023C (en) | 2004-07-30 | 2004-07-30 | Quick breeding technolgy for Renanthera imschootiana Rolfe |
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| CN 200410050909 CN1255023C (en) | 2004-07-30 | 2004-07-30 | Quick breeding technolgy for Renanthera imschootiana Rolfe |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101112174B (en) * | 2007-08-23 | 2010-08-18 | 浙江省农业科学院 | Tissue cultivation quick breeding method of kangarno paws |
| CN101584298B (en) * | 2009-06-02 | 2011-09-14 | 中国科学院华南植物园 | Test-tube breeding method for Nothodoritis germchit |
| CN102144536A (en) * | 2011-03-09 | 2011-08-10 | 中国科学院华南植物园 | Method for crossbreeding and aseptic sowing of renanthera coccinea |
| CN106489730B (en) * | 2016-09-30 | 2018-09-28 | 中国科学院华南植物园 | A kind of Herba Renantherae coccineae once-seedling forming quick propagating method and culture medium |
| CN107580890A (en) * | 2017-09-12 | 2018-01-16 | 中国科学院华南植物园 | A kind of Herba Renantherae coccineae cuttage rapid propagating method |
| CN107711500A (en) * | 2017-10-25 | 2018-02-23 | 黄海民 | A kind of breeding method of orchid seedling |
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