CN101637096A - Quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method - Google Patents
Quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method Download PDFInfo
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Abstract
The invention provides a quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method which belongs to a quick and high-efficiency plant seed seedling breeding method. The method comprises the following steps: (1) seed collection; (2) cleaning and sterilization of seeds; (3) release of seed dormancy at low temperature; (4) sterile germination of seeds; and (5) acclimatization and transplantation. The invention can enhance the seed germinating bud rate to more than 90% (enhanced by about 70%), and the seeding viability exceeds 90% (enhanced by about 60%); compared with the traditional method, the stocking time is shortened by about 2/3; the regular high-quality cephalotaxus breeding seedlings can be produced; and the invention provides technical support for the quick propagation and restoration protection of rare or endangered species of cephalotaxus hainanensis and the development and utilization of resources.
Description
Technical field
The present invention relates to the seed seedling quick efficient breeding method of a plant species, particularly quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method.
Background technology
Cephalotaxus mannii (Cephalotaxus hainanensis Li) is the Cephalotaxaceae plant, and another name cephalotaxus hainanensis, slender lobule bamboo comb China fir are perennial tall and big aiphyllium.Be the distinctive rare tree species of China, its distribution is very narrow, is detected in some areas such as Hainan and Guangdong, Yunnan.Studies show that the not only effective alkaloid of Cephalotaxus mannii is the highest in the Cephalotaxaceae, and curative effect is the most remarkable, is one of best anticancer seeds of generally acknowledging at present.These seeds are cut down repeatly because of material is good in addition, and population quantity is few in addition, are one of most important rare or endangered species of China.But this seeds natural resources is very limited, is difficult to satisfy growing demand.Because that these seeds are dioecism, seed production and germination rate is very low (the only tens of grain of annual every female plant, seed germination rate open-air about 10% (is that Hainan is difficult to satisfy because the Cephalotaxus mannii seed germination needs the low temperature breaking dormancy on the one hand; On the other hand the Cephalotaxus mannii seed very easy in the open air by animal gnaw, tradition is artificial sprouts about 20%); and the open-air and traditional germination method needs of Cephalotaxus mannii seed are about, and the time could Cheng Miao more than 1 year; be subjected to factor such as its ecological and artificial destruction in addition and cause the endangered (Wang Xianpu of Cephalotaxus mannii; Wang Yousheng. reason that Cephalotaxus mannii is in imminent danger and safeguard measure. GUIHAIA; 1994,14 (4): 369-372).Though also the someone studies the employing cottage propagation, but under the endangered situation of germ plasm resource, provide the difficulty of a large amount of nursery stocks very big by cuttage, do not satisfy the demand of pharmaceutical industry far away, and can cause bigger destruction the limited resources of Cephalotaxus mannii to Cephalotaxus mannii; And data shows that cuttage tender stem segments source directly has influence on cuttage growth of seedlings situation, and only deriving from the seedling that the young shoot cuttage on stem top produces can normal upright well grow; And stem top tender stem segments is drawn materials and can directly be destroyed the normal growth of this strain plant.So, cottage propagation can not promote substantially (Fu Wenying, Pan Xuefeng, Chen Xiuhui, Liu its people, Li Renzhu. the Cephalotaxus mannii cuttage and the experimental study of taking root. University Of Hainan's journal (natural science edition), 1997,15 (3): 239-244; Pan Xuefeng, Fu Wenying, Chen Xiuhui, Liu its people, Li Renzhu. the research of Cephalotaxus mannii cottage propagation. tropical agriculture science, 1996, (3): 25-30; Du Daolin, Fu Wenying. rare or endangered species Cephalotaxus mannii population protection bioecology. Hunan science and technology publishing house, 2003.11).Also the someone studies and adopts traditional method for tissue culture that Cephalotaxus mannii is carried out the seedling breeding in addition, but can not emerge substantially, perhaps emergence rate is very low, can not carry out application (Fu Wenying, Du Daolin, tally is equal. the inducing and cultivating of Cephalotaxus mannii callus. and Plant Physiology Communications, 2004,40 (1): 34-36; Li Zhiying, Wang Zhunian, Xu Li. the breeding fast of exsomatizing of Cephalotaxus mannii. Plant Physiology Communications, 2005,41 (5): 786).
Given this, the species conservation of Cephalotaxus mannii causes the great attention of botany research institution.So; the invention provides a kind of systems approach of directly rapidly and efficiently growing seedlings with the Cephalotaxus mannii seed; can realize improving seed germination bud ratio to 90% above (having improved about 70%); seedling survives and surpasses 90% (having improved about 60%); become seedling time ratio conventional method to shorten more than 2/3; and the high-quality Cephalotaxus mannii seedling of production neat and consistent; also utilized extremely limited Cephalotaxus mannii seed resource to greatest extent; the rare or endangered species Cephalotaxus mannii is fast numerous to provide technical guarantee with recovery and protection and resources development and utilization in order to solve, to satisfy this rare or endangered species seedling active demand of Cephalotaxus mannii.
Summary of the invention
The key technical problem that the present invention solves is directly rapidly and efficiently to grow seedlings with the Cephalotaxus mannii seed; improve more than the seed germination bud ratio to 90%; seedling survives and surpasses 90%; become seedling time ratio conventional method to shorten more than 2/3; and the high-quality Cephalotaxus mannii seedling of production neat and consistent, the rare or endangered species Cephalotaxus mannii is fast numerous to provide technical guarantee with recovery and protection and resources development and utilization in order to solve.
The technical problem to be solved in the present invention is to be brood body with the Cephalotaxus mannii seed, and a whole set of Cephalotaxus mannii seedling method for quickly breeding is provided.The technical scheme that addresses the above problem comprises the steps:
(1) seed collection: collect and grow near ripe or ripe Cephalotaxus mannii seed;
(2) cleaning and sterilization of seed: with Cephalotaxus mannii seed bath 1-3h., and clean up with distilled water, put into about 0.5-2.0% liquor natrii hypochloritis vacuum filtration about 3-8min that sterilizes after filter paper blots, sterile water washes 3-6 time repeatedly then;
(3) seed low temperature breaking dormancy: the good Cephalotaxus mannii seed of will sterilizing places 1-6 ℃ low-temperature preservation 1-3 week, removes Seed Dormancy;
(4) seed asepsis sprouting: the seed of breaking dormancy is put into the about 3-8min of about 0.5-2.0% liquor natrii hypochloritis vacuum filtration sterilization again, and sterile water washes 2-5 time repeatedly then; Then seed is inoculated on the following medium and cultivates: 0.5-1MS (Murashige and Skoog Stock medium)+0.01-0.1mgL
-16-BA (6-benzyl purine)+0-100mgL
-1Inositol+0.2-0.6% agar+1-5% sucrose+1-5% coconut milk, pH value 5.0-6.0; Place periodicity of illumination 16hd again behind the dark cultivation 2d
-1, intensity of illumination 2000-3000Lux cold light lamp under, continue in 25 ± 2 ℃ of the environmental temperatures to cultivate 15-25d, just obtain the seedling that root is arranged of new green, emergence rate surpasses 90%;
(5) domestication is transplanted: the good seedling that will take root moves to domestication chamber, intensity of illumination 10000-3000Lux, domestication 1-2 week, seedling is taken out from test tube, with warm water washing shoot root portion medium, use cold water flush again, transplant to volume ratio be coconut palm chaff: river sand=1-3: in 1 or frog stone: perlite: peat=1-3: 1-3: the 1 basin dress artificial substratum that mixes, every basin seedling puts transparent plastic bag, every 3 days sack is cut an osculum, and 9-12d goes basin seedling transparent plastic bag to move to hot-house culture; 1-2 moves into flowerpot respectively after week or a bag seedling is made in pack, and the matrix main component is cultivated 1-2 week again by fetched earth from open-air Cephalotaxus mannii group earth and humus, can change outdoor booth incubation growth over to; Seedling percent reaches more than 90%.
The invention has the beneficial effects as follows: directly rapidly and efficiently grow seedlings with the Cephalotaxus mannii seed; improve seed germination bud ratio to 90% above (having improved about 70%); seedling survives and surpasses 90% (having improved about 60%); become seedling time ratio conventional method to shorten more than 2/3; and the high-quality Cephalotaxus mannii seedling of production neat and consistent, the rare or endangered species Cephalotaxus mannii is fast numerous to provide technical guarantee with recovery and protection and resources development and utilization in order to solve.
Embodiment
The specific embodiment of the invention is illustrated below in conjunction with example: required various medicine and reagents all are commercially available in the medium, press the adapted of conventional method shown in the catalogue, by industry general.
Embodiment 1:
(1) seed collection: collect and grow near 20 in ripe or ripe Cephalotaxus mannii seed;
(2) cleaning and sterilization of seed: with Cephalotaxus mannii seed bath 1h., and clean up with distilled water, put into 0.5% liquor natrii hypochloritis's vacuum filtration sterilization 8min after filter paper blots, sterile water washes 3 times repeatedly then;
(3) seed low temperature breaking dormancy: the good Cephalotaxus mannii seed of will sterilizing places 1 week of about 1 ℃ low-temperature preservation, removes Seed Dormancy;
(4) seed germination: the seed of breaking dormancy is put into 0.5% liquor natrii hypochloritis's vacuum filtration sterilization 3min again, and sterile water washes 3 times repeatedly then; Then seed is inoculated on the following medium and cultivates: 0.5MS (Murashige and SkoogStock medium)+0.01mgL
-16-BA (6-benzyl purine)+0.2% agar+1% sucrose+1% coconut milk, pH value 5.0; Place periodicity of illumination 16hd again behind the dark cultivation 2d
-1, intensity of illumination 2000Lux cold light lamp under, continue to cultivate 15d in 25 ± 2 ℃ of the environmental temperatures, obtain the seedling that root is arranged 19 strains of new green, emergence rate surpasses 95%;
(5) domestication is transplanted: seedling is moved to domestication chamber, intensity of illumination 10000Lux, 1 week of domestication, seedling is taken out from test tube, with warm water washing shoot root portion medium, use cold water flush again, transplant to volume ratio be the coconut palm chaff: in the basin dress artificial substratum of river sand=mix at 1: 1, every basin seedling puts transparent plastic bag, every 3 days sack is cut an osculum, and 9d goes basin seedling transparent plastic bag to move to hot-house culture; Move into flowerpot after 1 week respectively and make the basin seedling, the matrix main component cultivated for 1 week again by fetched earth from open-air Cephalotaxus mannii group earth and humus, promptly changed outdoor booth incubation growth over to; Seedling percent reaches 100%.
Embodiment 2:
(1) seed collection: collect and grow near 20 in ripe or ripe Cephalotaxus mannii seed;
(2) cleaning and sterilization of seed: with Cephalotaxus mannii seed bath 3h, and clean up with distilled water, put into 2% liquor natrii hypochloritis's vacuum filtration sterilization 3min after filter paper blots, sterile water washes 6 times repeatedly then;
(3) seed low temperature breaking dormancy: the good Cephalotaxus mannii seed of will sterilizing places 2 weeks of about 6 ℃ low-temperature preservation, removes Seed Dormancy;
(4) seed germination: the seed of breaking dormancy is put into 2% liquor natrii hypochloritis's vacuum filtration sterilization 8min again, and sterile water washes 6 times repeatedly then; Then seed is inoculated on the following medium and cultivates: 1MS (Murashige and SkoogStock medium)+0.1mgL
-16-BA (6-benzyl purine)+100mgL
-1Inositol+0.6% agar+5% sucrose+5% coconut milk, pH value 6.0; Place periodicity of illumination 16hd again behind the dark cultivation 2d
-1, intensity of illumination 3000Lux cold light lamp under, continue to cultivate 25d in 25 ± 2 ℃ of the environmental temperatures, obtain the seedling that root is arranged 20 strains of new green, emergence rate 100%;
(5) domestication is transplanted: seedling is moved to domestication chamber, intensity of illumination 30000Lux, 2 weeks of domestication, seedling is taken out from test tube, with warm water washing shoot root portion medium, use cold water flush again, transplant to volume ratio be the coconut palm chaff: in the basin dress artificial substratum of river sand=mix at 3: 1, every basin seedling puts transparent plastic bag, every 3 days sack is cut an osculum, and 12d goes basin seedling transparent plastic bag to move to hot-house culture; Move into flowerpot after 2 weeks respectively and make the basin seedling, the matrix main component cultivated for 2 weeks again by fetched earth from open-air Cephalotaxus mannii group earth and humus, promptly changed outdoor booth incubation growth over to; Seedling percent reaches 100%.
Implement by technical method of the present invention, composition relative scale difference can cause the difference on rooting rate, the strong weak degree in the medium, and in a word, numerical value is more near optimum value, and result of implementation is unreasonable to be thought.
Claims (3)
1, quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method is characterized in that carrying out according to following step:
(1) seed collection: collect and grow near ripe or ripe Cephalotaxus mannii seed;
(2) cleaning and sterilization of seed: with Cephalotaxus mannii seed bath 1-3h., and clean up with distilled water, put into 0.5-2.0% liquor natrii hypochloritis vacuum filtration sterilization 3-8min after filter paper blots, sterile water washes 3-6 time repeatedly then;
(3) seed low temperature breaking dormancy: the good Cephalotaxus mannii seed of will sterilizing places 1-6 ℃ low-temperature preservation 1-3 week, removes Seed Dormancy;
(4) seed asepsis sprouting: the seed of breaking dormancy is put into about 0.5-2.0% liquor natrii hypochloritis vacuum filtration sterilization 3-8min again, and sterile water washes 2-5 time repeatedly then; Then seed is inoculated on the medium and cultivates; Place periodicity of illumination 16hd again behind the dark cultivation 2d
-1, intensity of illumination 2000-3000Lux cold light lamp under, continue to cultivate 15-25d in 25 ± 2 ℃ of the environmental temperatures;
(5) domestication is transplanted: the good seedling that will take root moves to domestication chamber, intensity of illumination 10000-3000Lux, domestication 1-2 week, seedling is taken out from test tube, with warm water washing shoot root portion medium, use cold water flush again, transplant in the basin dress artificial substratum, every basin seedling puts transparent plastic bag, every 3 days sack is cut an osculum, and 9-12d goes basin seedling transparent plastic bag to move to hot-house culture; 1-2 moves into flowerpot respectively after week or a bag seedling is made in pack, and the matrix main component is cultivated 1-2 week again by fetched earth from open-air Cephalotaxus mannii group earth and humus, can change outdoor booth incubation growth over to.
2, according to the described quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method of claim, it is characterized in that the composed as follows of medium: 0.5-1Murashige and Skoog Stock medium+0.01-0.1mgL
-16-benzyl purine+0-100mgL
-1Inositol+0.2-0.6% agar+1-5% sucrose+1-5% coconut milk, adjust pH 5.0-6.0.
3, according to the described quick and high-efficiency cephalotaxus hainanensis seed seedling breeding method of claim, it is characterized in that basin dress artificial substratum is composed as follows: count coconut palm chaff: river sand=1-3: 1 or frog stone: perlite: peat=1-3: 1-3: 1 with volume ratio.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103828680A (en) * | 2014-03-20 | 2014-06-04 | 中国热带农业科学院椰子研究所 | Rapid cultivation method for coconut seedlings |
| CN104186121A (en) * | 2014-08-08 | 2014-12-10 | 岭南园林股份有限公司 | One-time collecting and sowing method for seeds different in maturation periods |
| CN105830890A (en) * | 2016-04-08 | 2016-08-10 | 江西樟树天齐堂中药饮片有限公司 | Soilless seedling culturing and cultivation technology of gardenia jasminoides |
| CN105850430A (en) * | 2016-04-08 | 2016-08-17 | 江西樟树天齐堂中药饮片有限公司 | Fructus aurantii disease-resistant high-yield cultivation technology |
| CN105875146A (en) * | 2016-04-18 | 2016-08-24 | 杭州富阳飞博科技有限公司 | Vegetative propagation method of torreya jackii |
| CN108668558A (en) * | 2018-04-19 | 2018-10-19 | 边学静 | A method of abolishing Cephalotaxus mannii seed dormancy |
| CN109496653A (en) * | 2018-11-27 | 2019-03-22 | 山东省林业科学研究院 | A method of it is suitable for that extensive nursery quickly improves acer truncatum shoot cutting rooting rate |
| CN110463524A (en) * | 2019-09-10 | 2019-11-19 | 湖北理工学院 | A kind of method for culturing seedlings of copper weed |
| CN116138055A (en) * | 2023-04-06 | 2023-05-23 | 海南热带海洋学院 | Cephalotaxus hainanensis light induced high-pressure rooting propagation method |
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2009
- 2009-08-14 CN CN2009101842078A patent/CN101637096B/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103828680A (en) * | 2014-03-20 | 2014-06-04 | 中国热带农业科学院椰子研究所 | Rapid cultivation method for coconut seedlings |
| CN103828680B (en) * | 2014-03-20 | 2015-08-05 | 中国热带农业科学院椰子研究所 | A kind of fast breeding method of coconut seedling |
| CN104186121A (en) * | 2014-08-08 | 2014-12-10 | 岭南园林股份有限公司 | One-time collecting and sowing method for seeds different in maturation periods |
| CN105830890A (en) * | 2016-04-08 | 2016-08-10 | 江西樟树天齐堂中药饮片有限公司 | Soilless seedling culturing and cultivation technology of gardenia jasminoides |
| CN105850430A (en) * | 2016-04-08 | 2016-08-17 | 江西樟树天齐堂中药饮片有限公司 | Fructus aurantii disease-resistant high-yield cultivation technology |
| CN105875146A (en) * | 2016-04-18 | 2016-08-24 | 杭州富阳飞博科技有限公司 | Vegetative propagation method of torreya jackii |
| CN108668558A (en) * | 2018-04-19 | 2018-10-19 | 边学静 | A method of abolishing Cephalotaxus mannii seed dormancy |
| CN109496653A (en) * | 2018-11-27 | 2019-03-22 | 山东省林业科学研究院 | A method of it is suitable for that extensive nursery quickly improves acer truncatum shoot cutting rooting rate |
| CN110463524A (en) * | 2019-09-10 | 2019-11-19 | 湖北理工学院 | A kind of method for culturing seedlings of copper weed |
| CN110463524B (en) * | 2019-09-10 | 2022-03-22 | 湖北理工学院 | Seedling raising method for copperleaf flowers |
| CN116138055A (en) * | 2023-04-06 | 2023-05-23 | 海南热带海洋学院 | Cephalotaxus hainanensis light induced high-pressure rooting propagation method |
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