CN103548669B - Method for promoting asymbiotic germination of cymbidium ensifolium seeds - Google Patents
Method for promoting asymbiotic germination of cymbidium ensifolium seeds Download PDFInfo
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Abstract
The invention discloses a method for promoting asymbiotic germination of cymbidium ensifolium seeds. The cymbidium ensifolium seeds are obtained through artificial pollination; when successfully pollinated seeds grow for 300-360 days, the seeds are planted on a germination culture medium for germination; white or colorless transparent circular corms are formed after the seeds are geminated, and then are turned to be green rhizomes; the seeds start to germinate after being cultured for 25-30 days, and the germination rate is up to 50-75 percent when the seeds are cultured for 70 days; therefore, the aim of the method disclosed by the invention is fulfilled. The method has the advantages of high seed germination rate, short germination time, high rhizome quality, high actual operability, high application value and the like and has a very positive significance for improvement of cymbidium ensifolium seed germination rate and acceleration of cymbidium ensifolium seedling production.
Description
Technical field
The present invention relates to the germination method of traditional rare flower sword-leaved cymbidium seed, be specifically related to a kind of method of the sword-leaved cymbidium seed non-symbiosis germination based on aseptic seeding technology, there is significant application value.
Background technology
Sword-leaved cymbidium (Cymbidiu ensifolium) has another name called four seasons orchid, Xia Hui, sword a species of orchid, Qiu Lan, torch lily etc., for the orchid family (Ocrhidacea) Cymbidium (Cymbidium) perennial evergreen herbaceous plant, it is one of representational kind of most in state orchid, be distributed widely in Southeast Asian countries, domesticly mainly be distributed in the provinces and regions such as Fujian, Taiwan, Yunnan, Sichuan, Anhui, Guangxi and Guangdong, distribution is wider.The sword-leaved cymbidium florescence is unique, within 1 year, can bloom for several times and the florescence longer, be uniquely can remontant kind in numerous states orchid, sword-leaved cymbidium be many, unique, and flower pattern, pattern are all very abundant, and has very pleasant fragrance.Research finds sword-leaved cymbidium blade energy adsorption of dust, absorbs the toxic gas such as benzene and formaldehyde, is described as " SafeClean Utilities ".The same with other orchids; due to habitat destruction with excessively excavate; wild sword-leaved cymbidium is endangered; belong to the object of protection in " wild animals and plants Convention on International Trade in Endangered Species " (Convention on International Trade in Endangered species of Wild Fauna and Flora, CITES) annex I and be prohibited trade.
Sword-leaved cymbidium seed is very small, and embryo does not break up, and without endosperm or only there is a small amount of endosperm, natural germination rate is extremely low, and required time is long especially, generally takes 6-12 month or could sprout for more time.And by animal nutrition, non-symbiosis germination is carried out to Orchid Seeds, effectively can improve the germination rate of seed and shorten sprout time, being conducive to seeling industry obtains successfully, can produce good economic benefit.Therefore, promote that sword-leaved cymbidium seed non-symbiosis germination effectively can improve germination rate and reach the object of producing seedling fast by biotechnology.
Summary of the invention
The object of this invention is to provide that a kind of sword-leaved cymbidium seed germination rate is high, sprout time is short, the measured non-symbiosis germination method of root-like stock matter.
The present invention selects that ornamental value is high, market demand is comparatively large and adaptable sword-leaved cymbidium kind is material, utilizes aseptic seeding technology to carry out non-symbiosis germination research to its seed.Sword-leaved cymbidium seed is obtained by artificial pollination, when successful fruit growth of pollinating was by 300 ~ 360 days, planting seed is sprouted on germination medium, white or water white round bulb is formed after seed germination, change green root-like stock afterwards into, when cultivating 25 ~ 30 days, seed starts to sprout, and when 60 days, germination rate reaches 50 ~ 75%, thus achieves object of the present invention.
A kind of method promoting sword-leaved cymbidium seed non-symbiosis germination of the present invention, comprises following concrete steps:
(1) artificial pollination: carry out self-pollination in 1 ~ 2 day at flowering, mutually pollinates the pollen on different for same plant flower, removes lip simultaneously and carry out bagging process, preventing the interference of foreign pollen, remove bagging after 5 ~ 7 days, ripening rate 72.5 ~ 85%;
(2) selection of kernel maturing: the rear seed gathering different growth number of days of pollination is inoculated on germination medium, and carry out sprouting test, grow 300 ~ 360 days after selecting artificial pollination, capsule fully expands, and the fruit and seed that pericarp is dark green is optimal material; Described seed germination medium is: 1/6MS, NAA 6.0 ~ 10.0 mgL
-1, coconut milk 150 ~ 300 mL, active carbon 1.0 ~ 2.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2;
(3) seed presowing treatment: when capsule was grown by 300 ~ 360 days, whole fruit is cut and first cleans up with running water, then after soaking 2 ~ 3 minutes with the alcoholic solution that volume fraction is 70 ~ 75% successively, with the mercuric chloride solution soaking disinfection 15 ~ 20 minutes that mass fraction is 0.2%, Xian is rushed 4 ~ 6 times again with sterile water, blot the moisture of fruit surface with aseptic filter paper, longitudinally cut fruit, seed sterile gauze is wrapped at KT 0.1 ~ 0.3 mgmL
-1, IBA 0.1 ~ 1 mgmL
-1mixed solution in soak 4 ~ 6 minutes, then with aseptic washing 2 ~ 3 times;
(4) sowing and cultivating: the seed handled well is inoculated on germination medium, under the dark condition of temperature 30 ~ 32 DEG C, when cultivating 25 ~ 50 days, seed starts to expand the round bulb of formation water white transparency or white, and when 70 days, germination rate reaches 50 ~ 75%; Described seed germination medium contains: 1/6MS, NAA6.0 ~ 10.0 mgL
-1, coconut milk 150 ~ 300 mL, active carbon 1.0 ~ 2.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
Seed soaking solution described in step (3) is preferably: KT 0.3 mgmL
-1, IBA 0.2 mgmL
-1.
Described in step (2) and (4), seed germination medium is preferably: 1/6MS, NAA8.0 mgmL
-1, coconut milk 200 mL, active carbon 1.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
In described medium, NAA is 1-methyl α-naphthyl acetate.
Adopt this method, sword-leaved cymbidium seed germination required time is about 70 days, and germination rate reaches 50 ~ 75%.
Advantage of the present invention has: seed germination rate is high, sprout time is short, root-like stock quality is good, and actual operation is strong, using value advantages of higher, has very positive meaning to raising sword-leaved cymbidium seed germination efficiency and quickening sword-leaved cymbidium seeling industry.
Embodiment
Below in conjunction with embodiment, content of the present invention is further described, but is not limitation of the invention.
Embodiment 1
The non-symbiosis germination method of sword-leaved cymbidium kind treasured island fairy maiden's seed:
(1) artificial pollination: carry out self-pollination in 1 ~ 2 day at flowering, mutually pollinates the pollen on different for same plant flower, removes lip simultaneously and carry out bagging process, preventing the interference of foreign pollen, remove bagging after 5 days, ripening rate 79.5%;
(2) selection of kernel maturing: beginning in 120 days of pollinating gathers a fruit for every 30 days and carries out sprouting test, and germination medium is: 1/6MS, NAA8.0 mgL
-1, coconut milk 200 mL, active carbon 1.0 gL
-1, sucrose 20 g L
-1, agar 5.5 g L
-1, pH is 5.2, cultivate under the dark condition of temperature 25 ~ 28 DEG C, result with the seed of 360 days after artificial pollination for optimal material;
(3) seed presowing treatment: when capsule was grown by 360 days, whole fruit is cut and first cleans up with running water, then, after soaking 2 minutes with the alcoholic solution that volume fraction is 75% successively, be the mercuric chloride solution soaking disinfection 20 minutes of 0.2% with mass fraction, then rush Xian 6 times with sterile water.Blot the moisture of fruit surface with aseptic filter paper, longitudinally cut fruit, seed sterile gauze is wrapped at KT 0.3 mgmL
-1, IBA0.2 mg mL
-1solution in soak 5 minutes, then with aseptic washing 3 times;
(4) sowing and cultivating: the seed handled well is inoculated on germination medium, under the dark condition of temperature 30 ~ 32 DEG C, when cultivating 25 ~ 30 days, seed starts to expand the water white or white round bulb of formation, and when 70 days, germination rate reaches 75%; Described seed germination medium is: 1/6MS, NAA8.0 mgL
-1, coconut milk 200 mL, active carbon 1.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
Embodiment 2
The non-symbiosis germination method of sword-leaved cymbidium kind lotus element seed:
(1) artificial pollination: carry out self-pollination in 1 ~ 2 day at flowering, mutually pollinates the pollen on different for same plant flower, removes lip simultaneously and carry out bagging process, preventing the interference of foreign pollen, remove bagging after 5 days, ripening rate 85%;
(2) selection of kernel maturing: beginning in 120 days of pollinating gathers a fruit for every 30 days and carries out sprouting test, and germination medium is: 1/6MS, NAA6.0 mgL
-1, coconut milk 150 mL, active carbon 1.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2, cultivate under the dark condition of temperature 25 ~ 28 DEG C, result with the seed of 300 days after artificial pollination for optimal material;
(3) seed presowing treatment: when capsule was grown by 300 days, whole fruit is cut and first cleans up with running water, then, after soaking 2 minutes with the alcoholic solution that volume fraction is 70% successively, be the mercuric chloride solution soaking disinfection 15 minutes of 0.2% with mass fraction, then rush Xian 6 times with sterile water.Blot the moisture of fruit surface with aseptic filter paper, longitudinally cut fruit, seed sterile gauze is wrapped at KT 0.2 mgmL
-1, IBA 1.0 mg mL
-1solution in soak 5 minutes, then with aseptic washing 3 times;
(4) sowing and cultivating: the seed handled well is inoculated on germination medium, under the dark condition of temperature 30 ~ 32 DEG C, when cultivating 40 ~ 50 days, seed starts to expand the water white or white round bulb of formation, and when 70 days, germination rate reaches 50%; Described seed germination medium is: 1/6MS, NAA6.0 mgL
-1, coconut milk 150 mL, active carbon 1.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
Embodiment 3
The non-symbiosis germination method of sword-leaved cymbidium kind seven fairy maiden seed:
(1) artificial pollination: carry out self-pollination in 1 ~ 2 day at flowering, mutually pollinates the pollen on different for same plant flower, removes lip simultaneously and carry out bagging process, preventing the interference of foreign pollen, remove bagging after 5 days, ripening rate 72.5%;
(2) selection of kernel maturing: beginning in 120 days of pollinating gathers a fruit for every 30 days and carries out sprouting test, and germination medium is: 1/6MS, NAA10.0 mgL
-1, coconut milk 300 mL, active carbon 2.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2, cultivate under the dark condition of temperature 25 ~ 28 DEG C, result with the seed of 300 days after artificial pollination for optimal material;
(3) seed presowing treatment: when capsule was grown by 300 days, whole fruit is cut and first cleans up with running water, then, after soaking 2 minutes with the alcoholic solution that volume fraction is 70 successively, be the mercuric chloride solution soaking disinfection 15 minutes of 0.2% with mass fraction, then rush Xian 6 times with sterile water.Blot the moisture of fruit surface with aseptic filter paper, longitudinally cut fruit, seed sterile gauze is wrapped at KT 0.1 mgmL
-1, IBA 0.1 mgmL
-1solution in soak 5 minutes, then with aseptic washing 3 times;
(4) sowing and cultivating: the seed handled well is inoculated on germination medium, under the dark condition of temperature 30 ~ 32 DEG C, when cultivating 30 ~ 50 days, seed starts to expand the water white or white round bulb of formation, and when 70 days, germination rate reaches 56.7%; Described seed germination medium is: 1/6MS, NAA10.0 mgL
-1, coconut milk 300 mL, active carbon 2.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
Claims (3)
1. promote a method for sword-leaved cymbidium seed non-symbiosis germination, it is characterized in that comprising the following steps:
(1) artificial pollination: carry out self-pollination in 1 ~ 2 day at flowering, mutually pollinates the pollen on different for same plant flower, removes lip simultaneously and carry out bagging process, preventing the interference of foreign pollen, remove bagging after 5 ~ 7 days;
(2) selection of kernel maturing: the rear seed gathering different growth number of days of pollination is inoculated on germination medium, and carry out sprouting test, grow 300 ~ 360 days after selecting artificial pollination, capsule fully expands, and the fruit and seed that pericarp is dark green is optimal material; Described seed germination medium is: 1/6MS, NAA 6.0 ~ 10.0 mgL
-1, coconut milk 150 ~ 300 mL, active carbon 1.0 ~ 2.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2;
(3) seed presowing treatment: when capsule was grown by 300 ~ 360 days, whole fruit is cut and first cleans up with running water, then after soaking 2 ~ 3 minutes with the alcoholic solution that volume fraction is 70 ~ 75% successively, with the mercuric chloride solution soaking disinfection 15 ~ 20 minutes that mass fraction is 0.2%, Xian is rushed 4 ~ 6 times again with sterile water, blot the moisture of fruit surface with aseptic filter paper, longitudinally cut fruit, seed sterile gauze is wrapped at KT 0.1 ~ 0.3 mgmL
-1, IBA 0.1 ~ 1 mgmL
-1mixed solution in soak 4 ~ 6 minutes, then with aseptic washing 2 ~ 3 times;
(4) sowing and cultivating: the seed handled well is inoculated on germination medium, under the dark condition of temperature 30 ~ 32 DEG C, when cultivating 25 ~ 50 days, seed starts to expand the round bulb of formation water white transparency or white; Described seed germination medium is: 1/6MS, NAA6.0 ~ 10.0 mgL
-1, coconut milk 150 ~ 300 mL, active carbon 1.0 ~ 2.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
2. method according to claim 1, is characterized in that: described in step (3), seed presowing treatment solution is: KT 0.3 mgmL
-1, IBA 0.2 mgmL
-1mixed solution.
3. method according to claim 1, is characterized in that: described in step (2) and (4), seed germination medium is: 1/6MS, NAA8.0 mgL
-1, coconut milk 200 mL, active carbon 1.0 gL
-1, sucrose 20 gL
-1, agar 5.5 gL
-1, pH is 5.2.
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| CN104472487A (en) * | 2014-11-17 | 2015-04-01 | 广西源隆科技有限公司 | Chinese orchid root growth promoting agent and preparation method thereof |
| CN107484461A (en) * | 2017-09-12 | 2017-12-19 | 中国科学院华南植物园 | A kind of method that the pocket orchid seed germination rate for improving cryopreservation is handled using sodium hypochlorite |
Citations (3)
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| CN101816283A (en) * | 2010-05-05 | 2010-09-01 | 浙江大学 | Method for hybridizing cymbidium goeringii, aseptically sowing seeds and raising seedlings |
| CN102726288A (en) * | 2012-07-22 | 2012-10-17 | 云南农业大学 | Method for culturing and rapidly propagating large-flowered cymbidiums |
| CN103299911A (en) * | 2013-07-16 | 2013-09-18 | 葛建 | Method for obtaining virus-free seedlings of pure cymbidium efficiently |
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| KR20110074071A (en) * | 2009-12-24 | 2011-06-30 | 강경원 | New breed of cymbidium, oriental gold mine |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101816283A (en) * | 2010-05-05 | 2010-09-01 | 浙江大学 | Method for hybridizing cymbidium goeringii, aseptically sowing seeds and raising seedlings |
| CN102726288A (en) * | 2012-07-22 | 2012-10-17 | 云南农业大学 | Method for culturing and rapidly propagating large-flowered cymbidiums |
| CN103299911A (en) * | 2013-07-16 | 2013-09-18 | 葛建 | Method for obtaining virus-free seedlings of pure cymbidium efficiently |
Non-Patent Citations (3)
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| MICROPROPAGATION OF CYMBIDIUM ENSIFOLIUM VAR. MISERICORS THROUGH CALLUS-DERIVED RHIZOMES;CHEN CHANG et.al.,;《In Vitro Cell. Dev. Biol.--Plant》;20001231;第36卷;第517-520页 * |
| 兰科植物种子的非共生萌发研究进展;朱泉等;《江苏农业科学》;20091231(第4期);第2.3节 * |
| 多花兰种子无菌萌发及离体快繁研究;唐凤鸾等;《广西植物》;20121130;第32卷(第6期);第793-799页 * |
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