CN1240268C - Tissue cultivation quick breeding method of grass cherry blossom - Google Patents
Tissue cultivation quick breeding method of grass cherry blossom Download PDFInfo
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- CN1240268C CN1240268C CN 200310109643 CN200310109643A CN1240268C CN 1240268 C CN1240268 C CN 1240268C CN 200310109643 CN200310109643 CN 200310109643 CN 200310109643 A CN200310109643 A CN 200310109643A CN 1240268 C CN1240268 C CN 1240268C
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- 238000009395 breeding Methods 0.000 title abstract description 13
- 244000025254 Cannabis sativa Species 0.000 title abstract description 12
- 241000167854 Bourreria succulenta Species 0.000 title abstract 4
- 235000019693 cherries Nutrition 0.000 title abstract 4
- 238000000034 method Methods 0.000 claims abstract description 15
- 241000196324 Embryophyta Species 0.000 claims abstract description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 6
- 235000014001 Prunus serrulata Nutrition 0.000 claims description 20
- 239000006870 ms-medium Substances 0.000 claims description 18
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims description 17
- 239000002609 medium Substances 0.000 claims description 12
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 claims description 10
- 229930192334 Auxin Natural products 0.000 claims description 10
- 239000002363 auxin Substances 0.000 claims description 10
- PVRBGBGMDLPYKG-UHFFFAOYSA-N 6-benzyl-7h-purine Chemical compound N=1C=NC=2N=CNC=2C=1CC1=CC=CC=C1 PVRBGBGMDLPYKG-UHFFFAOYSA-N 0.000 claims description 9
- 238000012258 culturing Methods 0.000 claims description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 8
- 229930006000 Sucrose Natural products 0.000 claims description 8
- 235000013681 dietary sucrose Nutrition 0.000 claims description 8
- 230000001939 inductive effect Effects 0.000 claims description 8
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 229930191978 Gibberellin Natural products 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 4
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims description 4
- 239000003448 gibberellin Substances 0.000 claims description 4
- 239000011159 matrix material Substances 0.000 claims description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 235000019698 starch Nutrition 0.000 claims description 4
- 239000008107 starch Substances 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 3
- 229940064880 inositol 100 mg Drugs 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
- 235000015097 nutrients Nutrition 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 2
- 238000013138 pruning Methods 0.000 claims description 2
- 241000392970 Prunus serrulata Species 0.000 claims 2
- 230000001488 breeding effect Effects 0.000 abstract description 10
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000012364 cultivation method Methods 0.000 abstract description 2
- 238000009776 industrial production Methods 0.000 abstract 1
- 238000011031 large-scale manufacturing process Methods 0.000 abstract 1
- 230000035755 proliferation Effects 0.000 abstract 1
- 238000002054 transplantation Methods 0.000 abstract 1
- 244000141698 Prunus lannesiana Species 0.000 description 19
- 230000004069 differentiation Effects 0.000 description 5
- 229920002261 Corn starch Polymers 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000008120 corn starch Substances 0.000 description 3
- 238000012136 culture method Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 241000283986 Lepus Species 0.000 description 1
- 244000304321 Phlox drummondii Species 0.000 description 1
- 241001479570 Phlox subulata Species 0.000 description 1
- 238000010923 batch production Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention provides a quick breeding method for the tissue cultivation of grass cherry blossom, which is used for factory production to satisfy the market demand. The tissue cultivation of the grass cherry blossom comprises the steps of disinfection of stem apexes, inducement cultivation, proliferation cultivation, rootage cultivation and transplantation field planting. The grass cherry blossom is bred by a plant tissue cultivation method, so a large quantity of foreign exchange is saved, and the method is not influenced by external conditions. The method can be carried out in four seasons, occupied land for breeding seedlings is economized, and the production cost is reduced. All merits of a parent body are maintained, and the inheritable characteristic is stable. A large quantity of superior test tube seedlings can be formed in a short period by using the method, and the quick breeding method is suitable for large-scale production and industrial production.
Description
Technical field
The present invention relates to the tissue culture quick propagation culturing method of careless oriental cherry, belong to artificial propagation and the cultivation method technical field of plant.
Background technology
Grass oriental cherry (P.subulata L.) is born section's drummond phlox for flower and is belonged to perennial herb.Originate in Japan, a plurality of kinds such as adularescent, pink colour, pink, purple.Grass oriental cherry happiness light, cold-resistant heat-resisting again is adapted at the northern area of China growth, has and blooms early, happens few adolescence in early spring; flower is various when containing flower, and the florescence is long, and ornamental value is high; and characteristics such as the four seasons are evergreen, be a kind of good landscape ground by flowers, can be widely used in afforestation project.The grass oriental cherry mainly relies on import seminal propagation in China, and reproduction rate is very low.
Summary of the invention
The object of the present invention is to provide a kind of careless oriental cherry tissue culture quick propagation culturing method, carry out batch production production to meet the need of market.
Technical scheme of the present invention is achieved in that this careless oriental cherry tissue culture quick propagation culturing method, it is characterized in that:
A, stem apex sterilization: adult careless oriental cherry plant stem apex of clip or stem section, with flowing water flushing 1-2 hour, with mercuric chloride 0.1% sterilization 3-5 minute, aseptic water washing 3-5 time was drawn excessive moisture with filter paper then;
B, inducing culture: will wash the stem apex or the stem section that are cut into 0.3 centimetre of band leaf to clear material pruning, be inoculated in the inducing culture of following ratio: the MS medium of 2368.22 mg/litre is a minimal medium, add auxin 0.5-1.0 mg/litre, place culturing room to cultivate and grow up to test-tube plantlet more than 30 days;
C, enrichment culture: will cultivate the test-tube plantlet stem section more than 0.5 centimetre, be inoculated in the proliferated culture medium according to following ratio: the MS medium of 2368.22 mg/litre is a minimal medium, add inositol 100 mg/litre, auxin 1-2 mg/litre, S 3307 0.2 mg/litre, table sugar 30 grams per liters, starch 60 grams per liters carried out enrichment culture more than 25 days;
D, culture of rootage: the bud test-tube plantlet of growing thickly that clip is about 2.5 centimetres, be inoculated in the root media according to following ratio: the MS medium of 1184.11 mg/litre is a minimal medium, add auxin 0.1-2 mg/litre, table sugar 15 grams per liters, starch 60 grams per liters carried out culture of rootage 15-20 days;
E, transplant planting: with whole plant flush away root medium, be transplanted in the nutrient matrix, temperature 22-25 ℃, humidity 80%, field planting is in the flower nursery when waiting to grow more than 10 centimetres.
Described careless oriental cherry quick breeding by group culture method step B, C, D medium-height grass oriental cherry group are trained the auxin that relates in the described medium and are comprised 6-furfuryl group purine, 6-benzyl purine, gibberellin, indolebutyric acid, heteroauxin, methyl wherein one or more.
Described careless oriental cherry quick breeding by group culture method step B medium-height grass oriental cherry group is trained the auxin that relates in the described inducing culture and is comprised 6-furfuryl group purine or 6-benzyl purine 0.5-1.0 mg/litre.
Described careless oriental cherry quick breeding by group culture method step C medium-height grass oriental cherry group is trained the auxin that relates in the described proliferated culture medium and is comprised 6-benzyl purine 0.5-2 mg/litre, gibberellin 0.5-1 mg/litre.
Described careless oriental cherry tissue culture quick propagation culturing method is characterized in that step D medium-height grass oriental cherry group trains the auxin that relates in the described root media and comprise indolebutyric acid 0.1-1 mg/litre or heteroauxin 0.1-1 mg/litre or methyl 0.1-0.5 mg/litre.
Adopt the method for Plant Tissue Breeding to breed careless oriental cherry; adopt few resource to carry out the batch breeding of genetic stability; individual plant bud reproduction coefficient reaches more than 10, and rooting rate and transplanting survival rate all reach 100%, for improved seeds scale breeding provides the otherwise effective technique approach.
Embodiment
Breeding example explanation the specific embodiment of the present invention below in conjunction with careless oriental cherry group training.
Embodiment 1
1, test-tube plantlet inducing culture: stem apex, the stem section of the adult careless oriental cherry of clip, flowing water flushing 1-2 hour; Add mercuric chloride 0.1% sterilization 3-5 minute; Use aseptic water washing 3-5 time then, aseptic filter paper blots surface moisture, be cut into band base of leaf section or stem apex about 0.3 centimetre, put on the inducing culture: 2368.22 mg/litre MS add 6-benzyl purine 1 mg/litre, place culturing room to cultivate more than 30 days, grow up to test-tube plantlet, the test-tube plantlet inductivity is more than 95%.
2, the enrichment culture of test-tube plantlet: change about 0.5 centimetre stem apex or stem section over to proliferated culture medium, 2368.22 the MS medium of mg/litre, add 6-benzyl purine 2 mg/litre, inositol 100 mg/litre, S 3307 0.2 mg/litre, table sugar 30 grams per liters, corn starch 60 grams per liters, cultivate more than 25 days height of seedling 2-4 centimetre.Repeat same process, carry out the breeding of a large amount of test-tube plantlets, a month reproduction coefficient reaches 8-16.
3, the culture of rootage of test-tube plantlet: get the propagation test-tube plantlet (the bud seedling of growing thickly) about 2.5 centimetres, the MS medium of 1184.11 mg/litre is gone in switching, add indolebutyric acid or heteroauxin 0.5 mg/litre, 15 grams per liter table sugar, 50 grams per liter corn starchs the root original hase occurred in 7 days, grew complete root about 15 days, the long 0.2-0.5 of root centimetre, rooting rate 100%.Carry out test-tube seedling transplanting about 20 days, transplanting survival rate 100%.
4, whole plant is taken out from test tube, flush away root medium is transplanted in the nutrient matrix.Grow new blade about 5 days, spray nutritious liquor more than 15 days.When treating that seedling grows to 10 centimetres of left and right sides, carry out field planting.
5, at first prepare the loose soil of spending, with about 10 centimetres, the matrix seedling of robust growth, field planting is to the flower nursery.Under the normal management condition, transplanting survival rate 100%.Transplant and bloomed in 1 year, just spend generally less.
Embodiment 2
Repeat embodiment 1 operating process, the test-tube plantlet inducing culture is: the MS medium of 2368.22 mg/litre, 6-furfuryl group purine 1 mg/litre, S 3307 0.2 mg/litre, table sugar 30 grams per liters, corn starch 60 grams per liters; Bud propagation number is 6-10, and the bud differentiation rate is more than 90%.
Test-tube plantlet changes in the proliferated culture medium, and proliferated culture medium is: the MS medium of 2368.22 mg/litre, 6-benzyl purine 2 mg/litre, gibberellin 0.5-1 mg/litre, S 3307 0.2 mg/litre, table sugar 30 grams per liters, corn starch 60 grams per liters; Cultivate more than 25 days, height of seedling 1-4 centimetre, bud differentiation number reaches 8-20, differentiation rate 100%.
Below operation is with example 1.
Embodiment 3
Repeat embodiment 1 operating process, in the test-tube plantlet breeding, adopt following medium and result as follows:
2368.22 the MS medium of mg/litre adds 6-benzyl purine 1.25 mg/litre, bud differentiation number 6-13;
2368.22 the MS medium of mg/litre, 6-furfuryl group purine 1.5 mg/litre, bud differentiation number 6-14;
In rooting process, adopt following medium, the result is as follows:
1184.11 the MS medium of mg/litre adds indolebutyric acid 0.1 mg/litre, methyl 0.1 mg/litre, rooting rate 43%;
1184.11 the MS medium of mg/litre adds indolebutyric acid 0.2 mg/litre, rooting rate 58%;
1184.11 the MS medium of mg/litre adds indolebutyric acid 0.3 mg/litre, rooting rate 78%;
1184.11 the MS medium of mg/litre adds heteroauxin 0.2 mg/litre, rooting rate 55%;
1184.11 the MS medium of mg/litre adds heteroauxin 0.3 mg/litre, rooting rate 73%;
1184.11 the MS medium of mg/litre adds heteroauxin 0.4 mg/litre, rooting rate 91%.
Claims (1)
1, careless oriental cherry tissue culture quick propagation culturing method is characterized in that:
A, stem apex sterilization: adult careless oriental cherry plant stem apex of clip or stem section, with flowing water flushing 1-2 hour, with mercuric chloride 0.1% sterilization 3-5 minute, aseptic water washing 3-5 time was drawn excessive moisture with filter paper then;
B, inducing culture: will wash the stem apex or the stem section that are cut into 0.3 centimetre of band leaf to clear material pruning, be inoculated in the inducing culture of following ratio: the MS medium of 2368.22 mg/litre is a minimal medium, add auxin 6-furfuryl group purine or 6-benzyl purine 0.5-1.0 mg/litre, place culturing room to cultivate and grow up to test-tube plantlet more than 30 days;
C, enrichment culture: will cultivate the test-tube plantlet stem section more than 0.5 centimetre, be inoculated in the proliferated culture medium according to following ratio: the MS medium of 2368.22 mg/litre is a minimal medium, add people's inositol 100 mg/litre, auxin 6-benzyl purine 0.5-2 mg/litre, gibberellin 0.5-1 mg/litre, S 3307 0.2 mg/litre, table sugar 30 grams per liters, starch 60 grams per liters carried out enrichment culture more than 25 days;
D, culture of rootage: the bud test-tube plantlet of growing thickly that clip is about 2.5 centimetres, be inoculated in the root media according to following ratio: the MS medium of 1184.11 mg/litre is a minimal medium, add people's auxin indolebutyric acid 0.1-1 mg/litre or heteroauxin 0.1-1 mg/litre or methyl 0.1-0.5 mg/litre, table sugar 15 grams per liters, starch 60 grams per liters carried out culture of rootage 15-20 days;
E, transplant planting: with whole plant flush away root medium, be transplanted in the nutrient matrix, temperature 22-25 ℃, humidity 80%, field planting is in the flower nursery when treating that seedling grows to more than 10 centimetres.
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CN 200310109643 CN1240268C (en) | 2003-11-06 | 2003-11-06 | Tissue cultivation quick breeding method of grass cherry blossom |
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CN 200310109643 CN1240268C (en) | 2003-11-06 | 2003-11-06 | Tissue cultivation quick breeding method of grass cherry blossom |
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CN1240268C true CN1240268C (en) | 2006-02-08 |
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Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
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CN101112173B (en) * | 2006-07-28 | 2010-08-25 | 中国矿业大学(北京) | Tissue culture propagating method for heavy metal super-enriched thlaspi caerulescens |
CN101278625B (en) * | 2007-04-06 | 2010-09-22 | 谭宾 | Technique for cultivating oriental cherry |
CN105010144A (en) * | 2015-07-28 | 2015-11-04 | 福建世纪金花科技有限公司 | Double cherry tissue culture propagation method |
CN111084107A (en) * | 2020-02-18 | 2020-05-01 | 美尚生态景观股份有限公司 | Method for inducing axillary buds of cerasus humilis and subculture proliferation culture |
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