CN109511549B - Method for inhibiting differentiation and reversion of protocorm in bletilla striata seed culture process - Google Patents
Method for inhibiting differentiation and reversion of protocorm in bletilla striata seed culture process Download PDFInfo
- Publication number
- CN109511549B CN109511549B CN201811412031.2A CN201811412031A CN109511549B CN 109511549 B CN109511549 B CN 109511549B CN 201811412031 A CN201811412031 A CN 201811412031A CN 109511549 B CN109511549 B CN 109511549B
- Authority
- CN
- China
- Prior art keywords
- protocorm
- culture
- bletilla striata
- auxin
- culture medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims abstract description 38
- 241001313857 Bletilla striata Species 0.000 title claims abstract description 29
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 10
- 238000011218 seed culture Methods 0.000 title claims abstract description 8
- 230000004069 differentiation Effects 0.000 title claims description 7
- 239000001963 growth medium Substances 0.000 claims abstract description 26
- 229930192334 Auxin Natural products 0.000 claims abstract description 17
- 239000002363 auxin Substances 0.000 claims abstract description 17
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000005983 Maleic hydrazide Substances 0.000 claims abstract description 14
- BGRDGMRNKXEXQD-UHFFFAOYSA-N Maleic hydrazide Chemical compound OC1=CC=C(O)N=N1 BGRDGMRNKXEXQD-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000009630 liquid culture Methods 0.000 claims abstract description 12
- 239000007787 solid Substances 0.000 claims abstract description 11
- 239000000243 solution Substances 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 210000002257 embryonic structure Anatomy 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 230000012010 growth Effects 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 7
- 230000005764 inhibitory process Effects 0.000 abstract description 7
- 238000012258 culturing Methods 0.000 abstract description 3
- 239000002373 plant growth inhibitor Substances 0.000 abstract description 3
- 230000005087 leaf formation Effects 0.000 abstract 1
- 210000001161 mammalian embryo Anatomy 0.000 description 8
- 239000003966 growth inhibitor Substances 0.000 description 6
- 239000003112 inhibitor Substances 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 5
- 229930006000 Sucrose Natural products 0.000 description 4
- 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 description 4
- 229960004793 sucrose Drugs 0.000 description 4
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 3
- 241001313855 Bletilla Species 0.000 description 3
- 241001505935 Phalaenopsis Species 0.000 description 3
- 230000032823 cell division Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 241001076416 Dendrobium tosaense Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 230000007226 seed germination Effects 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000002352 surface water Substances 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- SNVRDQORMVVQBI-UPHRSURJSA-N (z)-but-2-enedihydrazide Chemical compound NNC(=O)\C=C/C(=O)NN SNVRDQORMVVQBI-UPHRSURJSA-N 0.000 description 1
- YDDUMTOHNYZQPO-UHFFFAOYSA-N 1,3-bis{[(2E)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]oxy}-4,5-dihydroxycyclohexanecarboxylic acid Natural products OC1C(O)CC(C(O)=O)(OC(=O)C=CC=2C=C(O)C(O)=CC=2)CC1OC(=O)C=CC1=CC=C(O)C(O)=C1 YDDUMTOHNYZQPO-UHFFFAOYSA-N 0.000 description 1
- ZMZGFLUUZLELNE-UHFFFAOYSA-N 2,3,5-triiodobenzoic acid Chemical compound OC(=O)C1=CC(I)=CC(I)=C1I ZMZGFLUUZLELNE-UHFFFAOYSA-N 0.000 description 1
- ZKQDCIXGCQPQNV-UHFFFAOYSA-N Calcium hypochlorite Chemical compound [Ca+2].Cl[O-].Cl[O-] ZKQDCIXGCQPQNV-UHFFFAOYSA-N 0.000 description 1
- SITQVDJAXQSXSA-CEZRHVESSA-N Cynarin Natural products O[C@@H]1C[C@@](C[C@H](O)[C@H]1OC(=O)C=Cc2ccc(O)c(O)c2)(OC(=O)C=Cc3cccc(O)c3O)C(=O)O SITQVDJAXQSXSA-CEZRHVESSA-N 0.000 description 1
- YDDUMTOHNYZQPO-RVXRWRFUSA-N Cynarine Chemical compound O([C@@H]1C[C@@](C[C@H]([C@@H]1O)O)(OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)C(O)=O)C(=O)\C=C\C1=CC=C(O)C(O)=C1 YDDUMTOHNYZQPO-RVXRWRFUSA-N 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
- 240000008790 Musa x paradisiaca Species 0.000 description 1
- 235000018290 Musa x paradisiaca Nutrition 0.000 description 1
- 241000233855 Orchidaceae Species 0.000 description 1
- 241000219000 Populus Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000020415 coconut juice Nutrition 0.000 description 1
- 235000020197 coconut milk Nutrition 0.000 description 1
- 229950009125 cynarine Drugs 0.000 description 1
- YDDUMTOHNYZQPO-BKUKFAEQSA-N cynarine Natural products O[C@H]1C[C@@](C[C@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)(OC(=O)C=Cc3ccc(O)c(O)c3)C(=O)O YDDUMTOHNYZQPO-BKUKFAEQSA-N 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 210000002308 embryonic cell Anatomy 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 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 description 1
- 239000003448 gibberellin Substances 0.000 description 1
- 235000021552 granulated sugar Nutrition 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000006194 liquid suspension Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000037039 plant physiology Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000002786 root growth Effects 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a method for maintaining the growth and expansion of protocorms, inhibiting sprouting and leaf formation and reversing the inhibition to form leaves and grow roots in the bletilla striata seed culture process. Shake culturing bletilla striata seed with liquid culture medium without auxin, making seed turn green to generate apical meristem, adding plant growth inhibitor maleic hydrazide to obtain protocorm with relatively complete structure, transferring the protocorm to solid culture medium with auxin, standing for culture, and allowing the protocorm to leave and grow into young seedling. The invention has simple operation, low cost, ideal effect and easy popularization and application.
Description
Technical Field
The invention belongs to a plant seed seedling raising technology in the field of agriculture and forestry, and particularly relates to a method for maintaining the growth and expansion of protocorms, inhibiting the emergence of leaves and roots and reversing the inhibition to realize the emergence of leaves and roots in the culture process of bletilla seeds.
Background
Bletilla striata (Bletilla striata) belongs to Orchidaceae, has high medicinal value, and the fruit contains thousands of seeds, and the seeds are fine and are a group of undifferentiated embryonic cells. The color of cells of bletilla striata embryo is changed into light green after water absorption and expansion, a growing point (apical meristem) with vigorous cell division activity appears at one end of the seed embryo, a growing cone is formed, the growing point is expanded to form a protocorm, a leaf primordium is differentiated from the top of the protocorm and gradually develops into leaves (Zhang Yan, and the like, seed germination and seedling morphology of bletilla striata on different culture media, northwest plant bulletin, 2009,29(8): 1584-1589). The protocorm is a common material for in vitro preservation of plant germplasm, artificial seed production, propagation by using a bioreactor, transgenic operation and the like.
One seed of bletilla striata expands into an protocorm, the top of the protocorm only contains one bud, and the observation shows that when the bletilla striata seed expands into a tender protocorm, the apical meristem also differentiates into germs at the same time, and the leaf and the root are quickly pulled out. The fast appearance of leaf and root organs, the nutrient absorbed by protocorm and the nutrient synthesized by photosynthesis are used for sprouting and root growing, the self growth and expansion of protocorm are affected, the protocorm with complete structure can be seen in the culture bottle, the volume is small, and the growth and retention time is short (leaf and root growing can be realized quickly). Bletilla striata embryo expands, shoots and grows leaves, and radicle bulge root extends later (Zhang Jianxia et al, relation between bletilla striata embryo development and seed germination. 2005, subtropical botanics science 34(4):32-35), so leaves are the main organs that destroy the structural integrity of protocorm and make it not near-globular. The mature protocorm is accompanied by longer leaves, which seriously influences the operation and effect of manufacturing artificial seeds, expanding propagation by using a bioreactor, preserving germplasm and the like.
The bud and leaf of the seed have back-to-ground growth, and the root has ground-to-ground growth. In the liquid shake culture, the seeds or protocorms roll continuously, so that the directionality of the sprouting and root growing is disturbed, the sprouting and root growing speed is slowed down, the growth and expansion of the protocorms are not influenced, and the relative integrity of the structure is kept. Similar examples include significant differentiation of protocorm bud of Dendrobium officinale Kimura et Migo on solid medium, weak and small embryoid, insignificant differentiation of protocorm bud on suspension medium, uniform and strong embryoid (Sujiang et al. dynamics research on solid and liquid suspension culture of protocorm of Dendrobium officinale Kimura et Migo. J.N.G. agricultural science, 2007, 16 (4): 161-165). The protocorm of the phalaenopsis can be prevented from generating polar differentiation and inhibiting the growth of buds and roots by carrying out liquid shake culture on the protocorm of the phalaenopsis (Xuwenhua. liquid culture research on proliferation of the phalaenopsis. Yangtze university (own edition) agro paper, 2007,4(4): 71-72).
The plant growth inhibitor (plant growth inhibitor) can inhibit the growth of apical meristem, so that the plant loses apical dominance, has more lateral branches and small leaves, and can not be reversed by externally applied gibberellin, but can be reversed by externally applied auxin (see: the main edition of panzu, plant physiology, 7 th edition in 2012, advanced education publishers, page 232; the main edition of Shimingwang and Populus pistil, the safe use speed of common pesticides, page 2017, the chemical industry publishers, 387 + 388). Common inhibitors are triiodobenzoic acid and maleic hydrazide (maleic dihydrazide, cynarin). After the maleic hydrazide is externally applied to the plant body, the maleic hydrazide is mainly concentrated to vigorous growth tissues and accumulated in terminal buds to act on apical meristems to inhibit cell division (Shimingwang and Zhusu Mei are mainly compiled, and safety use guidance of pollution-free vegetable garden pesticides is provided in 2017, chemical industry publishers, page 292-.
The bletilla striata seeds can be cultured and expanded in a liquid culture medium without auxin in a shaking way, for example, the culture medium of patent application No. 201510759434.4 is 1/2MS + 3% of cane sugar + 10-20% of coconut juice milk +0.5-1.5mg/L of 6-BA, the culture medium of application No. 201510922087.2 is MS +0.5-2.0mg/L of TDZ +30g/L of white granulated sugar, Jiangbeli and the like (Jiangbeli and the like, the research on the bletilla striata seedling raising technology based on protocorm culture, Anhui agricultural science, 2018, 46(6):39-42.) the culture medium is/2 MS +100g/L of potato juice +20g/L of cane sugar. That is, no auxin is added during the process of the bletilla striata seeds developing into protocorms, and when the growth inhibitor is added into the culture medium, the effect of the inhibitor is not counteracted.
Disclosure of Invention
The invention particularly relates to a method for keeping the growth and the swelling of protocorms and inhibiting sprouting and root growth during the culture process of bletilla seeds so as to obtain mature protocorms with complete structures and a method for reversing the inhibition to ensure that the protocorms grow leaves and roots.
The method adopts seed liquid culture medium added with growth inhibitor maleic hydrazide for shake culture. The seeds swell by absorbing water, all cells of the whole embryo grow and have consistent division level before the apical meristem appears, if the maleic hydrazide is added, the embryos are uniformly distributed in the whole embryo after being absorbed, the further swelling of the embryo can be inhibited, so the maleic hydrazide is added into the culture solution when the seeds are pre-cultured to be green and the apical meristem appears, and the maleic hydrazide is transferred to the apical meristem with vigorous cell division after being absorbed to play a role in inhibiting the sprouting without obvious inhibition on other tissues.
The culture medium does not contain auxin in the process of the bletilla striata seeds developing into protocorms, and the inhibition effect of a growth inhibitor cannot be counteracted. The protocorm with complete structure is obtained by the treatment of the growth inhibitor, if further sprouting and root growing are needed, the inhibition effect of the inhibitor on the sprouting of the protocorm can be reversely eliminated by transferring the protocorm to a solid culture medium added with auxin for standing culture, and the protocorm can leave leaves and grow roots.
The technical scheme adopted by the invention is as follows:
dissolving growth inhibitor maleic hydrazide with 4% sodium hydroxide to obtain an aqueous solution, and filtering and sterilizing the aqueous solution for later use. Preparing liquid culture medium of bletilla striata seed without auxin, and conventionally performing high-pressure sterilization. Sterilizing and disinfecting the surfaces of bletilla striata fruits, digging the fruits in a sterile environment, shaking off the seeds to a culture solution, and shaking to culture a shaking table at a rotating speed of 80-120 r/min. The culture temperature is 25 + -2 deg.C, and the illumination time is 10-12 h/d. After 10-20 days (different due to the maturity of the seeds), when the seeds are expanded to green and apical meristem appears, the filtered and sterilized maleic hydrazide solution is injected into the culture solution in a sterile environment and is mixed uniformly, and the concentration is 2-20 mg/L. The seeds continue to shake and grow under the original culture condition until mature protocorms with a nearly spherical shape are formed. In order to make the protocorm leave and grow roots, the protocorm is washed twice by sterile water, surface water is absorbed by filter paper, and the protocorm is transferred to a solid culture medium added with auxin for conventional standing culture, wherein the auxin is NAA (naphthalene acetic acid), indolebutyric acid (IBA) or a mixture of the two, the concentration of the auxin is 0.2-1.0mg/L, and the protocorm leaves and grows roots into tender seedlings.
The invention has the beneficial effects that: the invention can obviously inhibit the seeds of bletilla striata from leaf-drawing and root-growing in the process of growing and expanding to protocorm to obtain mature protocorm with relatively complete structure, and has no obvious influence on the growing and expanding of the protocorm. The inhibiting effect of the growth inhibitor is easy to reverse artificially, and leaves and roots can be smoothly grown. The method has the advantages of simple operation, low cost, ideal effect and easy popularization and application.
Detailed Description
Example one
The method comprises the following steps:
1. washing rhizoma bletilla fruit with water, soaking in 70-75% ethanol in a clean bench for instant sterilization, soaking in 10% calcium hypochlorite solution for 20 min, and washing with sterile water for 2 times. The basic culture medium of the seed culture medium is MS + 20% of coconut milk (V/V) + 2.0% of cane sugar (W/V), the common high-pressure sterilization is carried out, bletilla striata fruits are dissected on a super clean workbench, the seeds are shaken off to the surface of the culture medium, the liquid culture medium is shake-cultured, and the rotating speed of a shaking table is 100 r/min. The culture temperature is 25 +/-2 ℃, and the illumination time is 12 h/d.
2, (1) conventional solid culture: the formula of the culture medium is that the minimal medium is added with agar 0.08% (W/V). (2) conventional liquid culture: the formula of the culture solution is a basic culture medium. (3) Method of the invention (conventional liquid culture + inhibitor): dissolving maleic hydrazide with 4% sodium hydroxide solution to obtain water solution, and filtering and sterilizing. The seed culture solution formula is a basic culture medium, the seeds begin to turn green after shaking culture for 16 days, apical meristem appears, maleic hydrazide solution which is filtered and sterilized in advance is injected into the culture solution in a super clean bench, the final concentration is 5mg/L, and shaking culture is continued.
3. The above 3 methods were performed until the protocorm was "mature", and 50 protocorms were randomly observed for each treatment.
4. And (3) washing the bletilla striata protocorm obtained by the method (2) and the method (3) twice in an aseptic manner, sucking the surface water by using filter paper, transferring the bletilla striata protocorm to a solid culture medium MS + NAA0.5mg/L + 10% banana puree (W/V) + sucrose 2.0% (W/V) + agar 0.08% (W/V) added with auxin NAA, standing and culturing, and observing the germination condition of the protocorm.
(II) results:
1. bletilla striata seeds obviously turn green and swell after being cultured in a liquid culture medium for 16 days, solid culture seeds turn green slightly later but sprout quickly and grow leaves and root, protocorms with complete structures are not seen all the time, and observation data are shown in the following table.
TABLE. three methods for culturing mature protocorm of bletilla striata seed
The conventional liquid culture (method 2) and the method of the invention (method 3) have the advantages that the seed expansion and development are faster, the seed culture lasts for 45 days, the size and the specification are similar, and the seed embryo expansion is 1 time larger than that of the seed embryo cultured for 60 days in a solid state.
Compared with the method (1), the method (3) obviously inhibits the average length of the longest leaves by 90 percent and reduces the number of the protocorms with leaves by 50 percent. The average length of the longest leaves of the protocorm obtained by the method (3) is one third of that of the protocorm obtained by the method (2), and the number of the protocorms with leaves is reduced by 29 percent compared with the method (2). The method of the present invention is almost never followed.
3. Transferring mature bletilla striata protocorm obtained by the methods (2) and (3) to a solid culture medium added with auxin, wherein the protocorm can normally grow leaves and root.
(III) conclusion:
the inhibitor adding method in the conventional liquid culture can obviously inhibit the bletilla striata seeds from leaf-drawing and root-growing in the process of expanding into protocorms, and has no obvious influence on the expansion and growth of the protocorms. The external application of auxin can reverse the inhibition of inhibitor, and protocorm can normally take out leaves and grow roots.
Claims (2)
1. A method for inhibiting differentiation and reversion of protocorms in bletilla striata seed culture process is characterized by comprising the following steps: inoculating bletilla striata seeds in a liquid culture medium without auxin for shake culture, adding maleic hydrazide into the culture solution when the seeds are expanded to generate apical meristem, continuing shake culture, and expanding and developing seed embryos into protocorms; washing protocorm with water, transferring to a solid culture medium added with auxin, standing for culture, and allowing the protocorm to leave leaves and grow roots;
the auxin is NAA, IBA, or their mixture, and the concentration is 0.2-1.0mg/L culture medium.
2. The method for inhibiting differentiation and reversion of protocorms during bletilla striata seed culture process according to claim 1, wherein: the maleic hydrazide is dissolved with 4% sodium hydroxide, and the aqueous solution is sterilized by filtration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811412031.2A CN109511549B (en) | 2018-11-25 | 2018-11-25 | Method for inhibiting differentiation and reversion of protocorm in bletilla striata seed culture process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811412031.2A CN109511549B (en) | 2018-11-25 | 2018-11-25 | Method for inhibiting differentiation and reversion of protocorm in bletilla striata seed culture process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109511549A CN109511549A (en) | 2019-03-26 |
| CN109511549B true CN109511549B (en) | 2021-03-16 |
Family
ID=65778688
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811412031.2A Expired - Fee Related CN109511549B (en) | 2018-11-25 | 2018-11-25 | Method for inhibiting differentiation and reversion of protocorm in bletilla striata seed culture process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109511549B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103548681A (en) * | 2013-10-31 | 2014-02-05 | 浙江农林大学天目学院 | Culture medium for rapidly inducing adventitious buds in tissue culture of dendrobium nobile |
| KR101434162B1 (en) * | 2012-09-28 | 2014-08-29 | 농업회사법인 (주) 그린아그로 | Method for inhibiting fructification of ginseng berry using maleic hydrazide and ginseng plant produced by the method |
| WO2015040493A2 (en) * | 2013-09-23 | 2015-03-26 | Centro De Investigacion Y De Estudios Avanzados Del Instituto Politecnico Nacional (Cinvestav) | Systems for cloning plants through asexual means |
| CN106416972A (en) * | 2016-08-29 | 2017-02-22 | 浙江金石生物科技有限公司 | Method for quickly cultivating bletilla striata seedlings |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104255496B (en) * | 2014-09-23 | 2016-03-16 | 江苏农林职业技术学院 | A kind of method of Fast-propagation bletilla striata test-tube plantlet |
-
2018
- 2018-11-25 CN CN201811412031.2A patent/CN109511549B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101434162B1 (en) * | 2012-09-28 | 2014-08-29 | 농업회사법인 (주) 그린아그로 | Method for inhibiting fructification of ginseng berry using maleic hydrazide and ginseng plant produced by the method |
| WO2015040493A2 (en) * | 2013-09-23 | 2015-03-26 | Centro De Investigacion Y De Estudios Avanzados Del Instituto Politecnico Nacional (Cinvestav) | Systems for cloning plants through asexual means |
| CN103548681A (en) * | 2013-10-31 | 2014-02-05 | 浙江农林大学天目学院 | Culture medium for rapidly inducing adventitious buds in tissue culture of dendrobium nobile |
| CN106416972A (en) * | 2016-08-29 | 2017-02-22 | 浙江金石生物科技有限公司 | Method for quickly cultivating bletilla striata seedlings |
Non-Patent Citations (4)
| Title |
|---|
| Plant Metabolic Studies of the Growth Regulator Maleic Hydrazide;Dieter Komossa等;《American Chemical Society》;19951231;第43卷(第10期);2713-2715 * |
| The mode of Action of Maleic Hydrazide：Inhibition of Growth;Larry D. Nooden;《PHYSIOLOGIA PLANTARUM》;19690228;第22卷;260-270 * |
| 不同因素对白芨原球茎和假鳞茎形成的影响;史俊等;《江苏农业科学》;20180925;第46卷(第18期);153-156 * |
| 植物根状茎发生机理的研究进展;王振兴等;《中国林副特产》;20100215(第1期);97-100 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109511549A (en) | 2019-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Anjanawe et al. | Effect of plant growth regulators and growth media on seed germination and growth vigour of papaya | |
| Amin et al. | In vitro response of apical bud explants from mature trees of jackfruit (Artocarpus heterophyllus) | |
| CN101176429A (en) | Culture medium composition adapting for blueberry test tube plantlet radication as well as method thereof | |
| CN104381131B (en) | A kind of Pinus tabuliformis somatic embryo occurs and plant regeneration method | |
| CN103782811A (en) | Watermelon tissue culture seedling test tube micro-grafting method | |
| CN107593454A (en) | A kind of anti-pine nematode masson pine somatic embryo occurs and plant regeneration method | |
| CN106718889A (en) | A kind of synchronized anti-pine nematode Japanese red pine somatic embryo high occurs and plant regeneration method | |
| CN111543137B (en) | Rose seed seedling raising method | |
| CN1154413C (en) | Amorphophallus rivieri group seedling-culturing batch production and cultivation technology | |
| CN111134019B (en) | Method for directly generating somatic embryos and regenerating plants of ilex denticulata | |
| RU2302106C1 (en) | Method for preparation of in vitro propagated strawberry (fragraria l) plants to ex vitro cultivation | |
| CN109511549B (en) | Method for inhibiting differentiation and reversion of protocorm in bletilla striata seed culture process | |
| KR101246135B1 (en) | Method for germination improvement of genus rosa spp. | |
| CN111587688A (en) | In-vitro preservation and breeding method of kiwi fruit resources | |
| WO2012116475A1 (en) | Seedling-raising and root-shaping agent for inserting into plate of rice seeding machine and application method thereof | |
| KR101390396B1 (en) | Multiplication method of monocotyledon by removing stipes and by treating low temperature | |
| CN112931226B (en) | Tissue culture rapid propagation method for alnus ferox | |
| CN109863998B (en) | Tissue culture method of tilia mongolica seedlings | |
| CN108668869B (en) | Hydroponic seedling culture method for arrowheads | |
| Lomtatidze et al. | Production of sapling material of blueberry in in vitro culture | |
| Gutiérrez-Nicolás et al. | Seed germination and in vitro propagation of Maytenus canariensis through regeneration of adventitious shoots from axillary and apical buds | |
| CN108967193A (en) | A kind of Potato Shoot-tips removing fast breeding technique method | |
| CN112704012B (en) | Preparation method of rejuvenating seedlings of Dioscorea polystachya | |
| CN117158320B (en) | Construction method of eustoma grandiflorum multi-variety somatic embryo efficient regeneration system | |
| KR102200935B1 (en) | Method for plant formation of jujube 'Bokjo' cultivar(Ziziphus jujuba Mill) using internode culture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20210316 |