CN103598092B - A kind of method that brownization of tree peony callus recovers - Google Patents
A kind of method that brownization of tree peony callus recovers Download PDFInfo
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- CN103598092B CN103598092B CN201310525612.8A CN201310525612A CN103598092B CN 103598092 B CN103598092 B CN 103598092B CN 201310525612 A CN201310525612 A CN 201310525612A CN 103598092 B CN103598092 B CN 103598092B
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- tree peony
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- 206010020649 Hyperkeratosis Diseases 0.000 title claims abstract description 56
- 240000005001 Paeonia suffruticosa Species 0.000 title claims abstract description 15
- 235000003889 Paeonia suffruticosa Nutrition 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title claims abstract description 12
- 239000012499 inoculation medium Substances 0.000 claims abstract description 8
- 239000001963 growth medium Substances 0.000 claims description 16
- 238000011081 inoculation Methods 0.000 claims description 16
- 229920001817 Agar Polymers 0.000 claims description 6
- 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 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 239000005720 sucrose Substances 0.000 claims description 6
- 241000196324 Embryophyta Species 0.000 abstract description 19
- 230000008929 regeneration Effects 0.000 abstract description 16
- 238000011069 regeneration method Methods 0.000 abstract description 16
- 230000006698 induction Effects 0.000 abstract description 7
- 230000015572 biosynthetic process Effects 0.000 abstract description 5
- 241000736199 Paeonia Species 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 238000011017 operating method Methods 0.000 description 3
- 235000006484 Paeonia officinalis Nutrition 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 238000012090 tissue culture technique Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000000763 evoking effect Effects 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of method that brownization of tree peony callus recovers, adopt the method that progressively reduces temperature to recover the callus of brownization, follow the synergy of controlling the hormone-content in inoculation medium simultaneously, the callus of brownization recovers significantly, carry out the induction of regeneration plant with the callus recovering, the regeneration plant of formation does not have difference with the plant of normal callus induction.
Description
Technical field
The present invention relates to a kind of method that brownization of tree peony callus recovers, belong to field of plant tissue culture technique.
Background technology
Tree peony is the perennial fallen leaves of Paeoniaceae Paeonia undershrubs, is China's tradition famous flower, splendid, aromatic, is deeply subject to liking of the people of the world. Tree peony breeding difficulty, mainly breeds by the mode of grafting for a long time, is difficult to so far realize the large-scale production of high quality seedling. At present, tissue culture technique for peony rises gradually, and the ways of regeneration that completes again regeneration plant by induction tree peony explant formation callus becomes fast the effectively shortcut of breeding peony plant; But tissue culture technique for peony is also immature at present, the inductivity of callus is not high, and in the callus forming, brownization problem is obvious. In the process that Browning control forms at evoked callus conventionally, in culture medium, add the material Browning controls such as active carbon to occur, for and tree peony that brownization is serious lower for callus induction rate, only inadequate by Browning control callus, if the callus of brownization can be reverted to normal callus and be used, be tantamount to further improve inductivity and the utilization rate of callus.
Summary of the inventionThe object of this invention is to provide a kind of method that brownization of tree peony callus recovers, the callus that brownization occurs is recovered, make it return to the go forward side by side generation of an one-step inducing regeneration plant of normal structure state.
The present invention is achieved through the following technical solutions: a kind of method that brownization of tree peony callus recovers, comprises the steps:
(1) inoculation for the first time: in gnotobasis, the tree peony callus of brownization is shifted in MS culture medium, cultivate 7d under 20 DEG C, 500-1000Lx;
(2) inoculation for the second time: the callus in step 1 is inoculated for the second time, cultivated 7d under 17 DEG C, full dark condition; (3) inoculation for the third time: the callus in step 2 is inoculated for the third time, cultivated 7d under 14 DEG C, full dark condition;
(4) the 4th inoculations: the callus in step 3 is carried out to the 4th inoculation, cultivate 7d under 10 DEG C, full dark condition; (5) the 5th inoculations: the callus in step 4 is carried out to the 5th inoculation, cultivate 7d under 15 DEG C, full dark condition.
Preferably, the inoculation medium described in described step 1-4 is MS culture medium, adds 40g/L sucrose and 7g/L agar in culture medium, and autoclaving is cooled to after room temperature stand-by.
Preferably, in described step 5, inoculation medium is MS culture medium, adds vitamin 20g/L sucrose and 7g/L agar in culture medium, and autoclaving is cooled to after room temperature stand-by.
Brownization of tree peony callus restoration methods of the present invention, adopt the method that progressively reduces temperature to recover the callus of brownization, simultaneously, the hormone that does not contain any kind and concentration in inoculation medium, avoid the side effect of any antioxidant, the callus of brownization recovers significantly, carries out the induction of regeneration plant with the callus recovering, the regeneration plant of formation with the plant of normal callus induction without any difference.
Detailed description of the inventionEmbodiment 1: the tree peony callus of brownization is shifted in MS culture medium in gnotobasis, cultivate 7d under 20 DEG C, 500-1000Lx; Above-mentioned callus is inoculated for the second time, under 17 DEG C, full dark condition, cultivated 7d; Above-mentioned callus is inoculated for the third time, under 14 DEG C, full dark condition, cultivated 7d; Callus in above-mentioned is carried out to the 4th inoculation, under 10 DEG C, full dark condition, cultivate 7d; Above-mentioned callus is carried out to the 5th inoculation, under 15 DEG C, full dark condition, cultivate 7d; Inoculation medium described in described step 1-4 is MS culture medium, adds 40g/L sucrose and 7g/L agar in culture medium; In described step 5, inoculation medium is MS culture medium, adds 20g/L sucrose and the equal autoclaving of 7g/L agar medium and be cooled to after room temperature stand-by in culture medium. Inoculate altogether the callus of 30 bottles of brownization, wherein 28 bottles are recovered normal, and these 28 bottles of callus are similar to the regeneration plant of normal callus induction, and the plant of formation is normal without deformity.
Embodiment 2: make step identical with embodiment 1, inoculate altogether the callus of 30 bottles of brownization, wherein 27 bottles are recovered normal, the regeneration plant that the callus of these 27 bottles of recoveries forms, the blade that blade forms regeneration plant than normal callus is roomy.
Embodiment 3: operating procedure is identical with embodiment 1, inoculates the callus of 30 bottles of brownization altogether, and wherein 24 bottles are recovered normal, the regeneration plant that the callus of these 24 bottles of recoveries forms, and plant height forms the short 1-3cm of regeneration plant than normal callus. Embodiment 4:
Operating procedure is identical with embodiment 1, inoculates altogether the callus of 30 bottles of brownization, and wherein 28 bottles are recovered normal, and the regeneration plant that the callus of these 28 bottles of recoveries forms is similar to normal callus formation regeneration plant.
Embodiment 5:
Operating procedure is identical with embodiment 1, inoculates altogether the callus of 30 bottles of brownization, and wherein 26 bottles are recovered normal, and the leaf color that the blade of the regeneration plant that the callus of these 26 bottles of recoveries forms forms regeneration plant than normal callus is bud green.
Above-mentioned example just, for explanation technical conceive of the present invention and technical characterstic, can not limit the scope of the invention with this. Equivalent transformation or modification that all essence according to the present invention is done, within all should being encompassed in protection scope of the present invention.
Claims (1)
1. the method that brownization of tree peony callus recovers, is characterized in that, comprises the steps:
(1) inoculation for the first time: in gnotobasis, the tree peony callus of brownization is shifted in MS culture medium, cultivate 7d under 20 DEG C, 500-1000Lx;
(2) inoculation for the second time: the callus in step 1 is inoculated for the second time, cultivated 7d under 17 DEG C, full dark condition;
(3) inoculation for the third time: the callus in step 2 is inoculated for the third time, cultivated 7d under 14 DEG C, full dark condition;
(4) the 4th inoculations: the callus in step 3 is carried out to the 4th inoculation, cultivate 7d under 10 DEG C, full dark condition;
(5) the 5th inoculations: the callus in step 4 is carried out to the 5th inoculation, cultivate 7d under 15 DEG C, full dark condition;
Inoculation medium described in above-mentioned steps 1-4 is MS culture medium, adds 40g/L sucrose and 7g/L agar in culture medium, and autoclaving is cooled to after room temperature stand-by;
In above-mentioned steps 5, inoculation medium is MS culture medium, adds 20g/L sucrose and 7g/L agar in culture medium, and autoclaving is cooled to after room temperature stand-by.
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| CN201310525612.8A CN103598092B (en) | 2013-10-31 | 2013-10-31 | A kind of method that brownization of tree peony callus recovers |
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| CN201310525612.8A CN103598092B (en) | 2013-10-31 | 2013-10-31 | A kind of method that brownization of tree peony callus recovers |
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| CN103598092A CN103598092A (en) | 2014-02-26 |
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| CN105648032B (en) * | 2016-02-04 | 2019-09-06 | 河南农业大学 | The gus gene histochemical stain method of transgenosis tree peony callus |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101548647A (en) * | 2009-05-11 | 2009-10-07 | 中国林业科学研究院林业研究所 | Tree peony callus induction method |
| CN103004602A (en) * | 2012-12-26 | 2013-04-03 | 安徽农业大学 | Method for suppressing browning of carya cathayensis sarg explant |
-
2013
- 2013-10-31 CN CN201310525612.8A patent/CN103598092B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101548647A (en) * | 2009-05-11 | 2009-10-07 | 中国林业科学研究院林业研究所 | Tree peony callus induction method |
| CN103004602A (en) * | 2012-12-26 | 2013-04-03 | 安徽农业大学 | Method for suppressing browning of carya cathayensis sarg explant |
Non-Patent Citations (2)
| Title |
|---|
| 牡丹离体培养中褐化问题的研究进展;牛佳佳等;《中国农学通报》;20091231;第25卷(第11期);第35页右栏最后两段及第36页左栏第2段 * |
| 牡丹组织培养中褐化的发生原因与防止方法的研究;张俊琦等;《沈阳农业大学学报》;20061231;第37卷(第5期);第720-725页 * |
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Address after: 225300 building 30, Xianglong home, Mingzhu street, medical high tech Zone, Taizhou City, Jiangsu Province Patentee after: Ma Genying Address before: 221000 No. 0111, Hardware Market Area 1, Jiefangnan Road Science and Technology City, Xuzhou City, Jiangsu Province Patentee before: Ma Genying |
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