AU2021107353A4 - A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application - Google Patents
A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application Download PDFInfo
- Publication number
- AU2021107353A4 AU2021107353A4 AU2021107353A AU2021107353A AU2021107353A4 AU 2021107353 A4 AU2021107353 A4 AU 2021107353A4 AU 2021107353 A AU2021107353 A AU 2021107353A AU 2021107353 A AU2021107353 A AU 2021107353A AU 2021107353 A4 AU2021107353 A4 AU 2021107353A4
- Authority
- AU
- Australia
- Prior art keywords
- medium
- preservation
- hosta
- plantlets
- vitro plantlets
- 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.)
- Ceased
Links
- 238000004321 preservation Methods 0.000 title claims abstract description 60
- 241001482471 Hosta plantaginea Species 0.000 title claims abstract description 40
- 238000000338 in vitro Methods 0.000 title claims abstract description 37
- 229930006000 Sucrose Natural products 0.000 claims abstract description 33
- 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 abstract description 33
- 239000005720 sucrose Substances 0.000 claims abstract description 33
- 229920001817 Agar Polymers 0.000 claims abstract description 29
- 239000008272 agar Substances 0.000 claims abstract description 29
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 24
- 229930195725 Mannitol Natural products 0.000 claims abstract description 24
- 239000000594 mannitol Substances 0.000 claims abstract description 24
- 235000010355 mannitol Nutrition 0.000 claims abstract description 24
- 230000012010 growth Effects 0.000 claims description 45
- 241000720945 Hosta Species 0.000 claims description 22
- 238000005286 illumination Methods 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- 239000002609 medium Substances 0.000 abstract description 42
- PWVXXGRKLHYWKM-UHFFFAOYSA-N 5-[2-(benzenesulfonyl)ethyl]-3-[(1-methylpyrrolidin-2-yl)methyl]-1h-indole Chemical compound CN1CCCC1CC(C1=C2)=CNC1=CC=C2CCS(=O)(=O)C1=CC=CC=C1 PWVXXGRKLHYWKM-UHFFFAOYSA-N 0.000 abstract description 17
- 230000007850 degeneration Effects 0.000 abstract description 8
- 239000006870 ms-medium Substances 0.000 abstract description 5
- 230000004083 survival effect Effects 0.000 abstract description 5
- JLIDBLDQVAYHNE-LXGGSRJLSA-N 2-cis-abscisic acid Chemical compound OC(=O)/C=C(/C)\C=C\C1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-LXGGSRJLSA-N 0.000 abstract description 2
- 238000004161 plant tissue culture Methods 0.000 abstract description 2
- JLIDBLDQVAYHNE-YKALOCIXSA-N Abscisic acid Natural products OC(=O)/C=C(/C)\C=C\[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-YKALOCIXSA-N 0.000 description 23
- 241000196324 Embryophyta Species 0.000 description 23
- 230000003111 delayed effect Effects 0.000 description 13
- 230000000694 effects Effects 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 229930002875 chlorophyll Natural products 0.000 description 5
- 235000019804 chlorophyll Nutrition 0.000 description 5
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000002335 preservative effect Effects 0.000 description 4
- 229930191978 Gibberellin Natural products 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- HLVPIMVSSMJFPS-UHFFFAOYSA-N abscisic acid beta-D-glucopyranosyl ester Natural products O1C(CO)C(O)C(O)C(O)C1OC(=O)C=C(C)C=CC1(O)C(C)=CC(=O)CC1(C)C HLVPIMVSSMJFPS-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 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 3
- 239000003448 gibberellin Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000003204 osmotic effect Effects 0.000 description 3
- 230000008635 plant growth Effects 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 239000004382 Amylase Substances 0.000 description 2
- 230000006819 RNA synthesis Effects 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- PZSMUPGANZGPBF-UHFFFAOYSA-N 4-[5-(dithiolan-3-yl)pentanoylamino]butanoic acid Chemical compound OC(=O)CCCNC(=O)CCCCC1CCSS1 PZSMUPGANZGPBF-UHFFFAOYSA-N 0.000 description 1
- SYMHUEFSSMBHJA-UHFFFAOYSA-N 6-methylpurine Chemical compound CC1=NC=NC2=C1NC=N2 SYMHUEFSSMBHJA-UHFFFAOYSA-N 0.000 description 1
- LPXQRXLUHJKZIE-UHFFFAOYSA-N 8-azaguanine Chemical compound NC1=NC(O)=C2NN=NC2=N1 LPXQRXLUHJKZIE-UHFFFAOYSA-N 0.000 description 1
- 229960005508 8-azaguanine Drugs 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 229940123457 Free radical scavenger Drugs 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 241000234280 Liliaceae Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 108010016634 Seed Storage Proteins Proteins 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 240000001949 Taraxacum officinale Species 0.000 description 1
- 235000005187 Taraxacum officinale ssp. officinale Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- FCRACOPGPMPSHN-UHFFFAOYSA-N desoxyabscisic acid Natural products OC(=O)C=C(C)C=CC1C(C)=CC(=O)CC1(C)C FCRACOPGPMPSHN-UHFFFAOYSA-N 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 229940125389 long-acting beta agonist Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 239000005648 plant growth regulator Substances 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 230000005068 transpiration Effects 0.000 description 1
- 230000009105 vegetative growth Effects 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 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
- A01H4/002—Culture media for tissue culture
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/12—Asparagaceae, e.g. Hosta
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)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
This invention discloses a preservation medium of Hosta plantaginea in vitro
plantlets. This invention belongs to the technical field of plant tissue culture. The
preservation medium of Hosta plantaginea in vitro plantlets in this invention is MS
medium supplemented with sucrose, mannitol, abscisic acid (ABA), chlormequat
chloride (CCC), and agar. The medium in this invention is used to preserve Hosta
plantaginea in vitro plantlets. Hosta plantaginea in vitro plantlets can be preserved
for six months and the survival rate of plantlets is 100%. What's more, the phenotype
of plantlets is stable and no degeneration or variation occurred after reculture. The
medium in this invention has low cost, overcomes the shortcomings of existing
preservation media such as short preservation duration and phenotypic variation after
preservation. It can be applied to the preservation of Hosta plantaginea germplasm
resources, and has great application value.
DRAWINGS
FIG.1
FIG. 2
1
Description
FIG.1
FIG. 2
A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application
TECHNICAL FIELD OF THE INVENTION The technical field of this invention is plant tissue culture. To be specific, it involves a preservation medium of Hostaplantagineain vitro plantlets and its application.
BACKGROUND OF THE RELATED ART Hosta plantaginea is a perennial herb of Hosta in liliaceae. It is a variety of famous foliage plant Hosta, which is highly complementary to light-absorbing flowers and plants and is an excellent variety for landscaping. Hosta plantaginea has dense clusters, oval-shaped to heart-shaped leaves, yellow stripes. There are so many cultivated varieties with great differences in leaf shape, leaf color and plant type. The green ones are green, the blue ones are faint, like fog and yarn, pretty and elegant, and the colorful leaves show magical gradient colors. In the great mass fervor of cultivating new varieties of Hosta in the world, Hosta plantaginea stands out among all Hosta varieties because of its unique leaf color. Therefore, Hosta plantagineahas become a wild flower and medicinal plant with great scientific research value and development prospect. Wild Hosta plants are often faced with the invasion of pathogenic bacteria, which causes leaves to become withered, base to become rotten and even the whole plant to die and also suffered from virus attacks. Hence, based on the long-term purpose of variety breeding, resources utilization and promoting the development of economy and industry of flowers and plants in China, the preservation of Hosta plant germplasm resources is of great importance. At present, the germplasm resources of Hosta plantaginea are mainly preserved in field nursery and in vitro genebanks. When preserved in the field, the germplasm can be easily affected by the environment and disappear. For example, underground rhizomes are easily invaded by viruses, making viruses accumulate in large quantities, leading to variety degeneration and yield reduction; in addition, urban land development and environmental change also seriously threaten the safe preservation of the germplasm resources of Hosta plantaginea. Although the preservation of in vitro plantlets has the advantages such as small floor area, not easily being affected by natural disasters and pathogenic bacteria, less investment in manpower and
financial resources, and convenient exchange and transportation of germplasm resources, it needs frequent subculture and renewal and faces the threat of degeneration or variation after subculture for many times.
SUMMARY OF THE INVENTION The purpose of this invention is to provide a preservation medium of Hosta plantagineain vitro plantlets and its application. The medium in this invention is a medium of Hosta plantaginea in vitro plantlets to make up for the deficiencies of the existing technologies such as short preservation period, and frequent subculture and renewal which can easily lead to the degeneration or variation of in vitro plantlets. According to the physiological and growth characteristics of Hosta plantaginea in vitro plantlets, this invention develops a medium especially suitable for preserving Hosta plantaginea in vitro plantlets. The formula of the medium is MS medium supplemented with mannitol, ABA, CCC and sucrose. Specifically, the medium in this invention is supplemented with MS+(502.5)g/L sucrose +(20±1)g/L mannitol +(1.00.05)mg/L ABA +19-42mg/L CCC +0.7 % agar
(pH 5.8). Preferably, the said medium is supplemented with MS+(50±2.0)g/L sucrose+(20±0.5)g/L mannitol+(1.0±0.04)mg/L ABA +20-40mg/L CCC+0.7% agar. More preferably, the said medium is supplemented with MS+(50±1.0)g/L sucrose+(20±0.2)g/L mannitol+(1.0±0.02)mg/L ABA +20-40mg/L CCC+0.7% agar. Most preferably, the said medium is supplemented with MS+50g/L sucrose+20g/L mannitol+1.Omg/L ABA +20mg/L CCC+0.7% agar (pH5.8); or The said culture medium is supplemented with MS+50g/L sucrose+20g/L mannitol+1.Omg/L ABA +40mg/L CCC+0.7% agar (pH 5.8). In the process of protoplast regeneration and culture, mannitol should be added to maintain the balance of osmotic pressure inside and outside the cytomembrane, increase the osmotic pressure, and prevent water from penetrating into the cell and causing cell rupture. Mannitol is mainly used to regulate the osmotic pressure of medium and play the role as a free radical scavenger. According to the invention, when the amount of mannitol in the medium of the invention is 1.9%-2.1%, especially 2% mannitol, combined with other components in the medium, the preservation period of Hosta plantaginea can be prolonged to 6 months, and the
survival rate of the preserved material is 100%, and the phenotype of the plant is stable without degeneration and variation after renewing culture. Abscisic acid (ABA) is a growth-inhibiting plant hormone which for its ability to promote leaves to fall off. Besides promoting leaves to fall off, it has other functions, such as making buds go dormant and promoting potatoes to form tubers. It also inhibits the prolongation of cells. It can improve the drought resistance and salt tolerance of plants and has extremely high value for developing and utilizing medium and low-yield fields, planting trees, greening deserts and so on. ABA is also an effective inhibitor to inhibit seed germination, so it can be used for seed storage to ensure the storage quality of seeds and fruits. In addition, ABA can also cause stomata in leaves to close rapidly, which can be used to keep flowers fresh, adjust flowering period and promote rooting. ABA plays a role in controlling nucleic acid and protein synthesis. It has been reported that ABA inhibits the synthesis of a amylase in barley grains and antagonizes gibberellin in this process. The inhibiting effect on enzymatic synthesis is similar to that of 8-azaguanine and 6-methylpurine which are inhibitors of RNA synthesis, indicating that the effect of ABA may be to inhibit the synthesis of RNA that determines the structure of a-amylase or to prevent RNA from binding to active enzyme units. ABA inhibits RNA synthesis in the leaves of dandelion and DNA synthesis in algae. ABA is sensitive to light and belongs to a compound of strong light degeneration. Chlormequat chloride (CCC) is a kind of quaternary ammonium salt plant growth regulator which can control the vegetative growth of plants (i.e., the growth of roots and leaves), promote the reproductive growth of plants (i.e., the growth of flowers and fruits), and improve the fruit setting rate of plants. CCC can regulate crop growth, promote tillering, and increase ears and production. After use, chlorophyll content increases, leaves' colors are dark green, photosynthesis is strengthened, leaves are thickened and root systems are developed. CCC hinders the biosynthesis of endogenous gibberellin, thus delaying cell elongation, making plants dwarfing, stems stout and intemodes shortened, and preventing plants from excessive growth and lodging. (The inhibitory effect of CCC on intemode elongation can be relieved by external administration of gibberellin.) CCC can improve the water absorbing capacity of root systems, obviously affect the accumulation of proline (which stabilizes the cell membrane) in plants, and is beneficial to improve the stress
resistance of plants, such as drought resistance, cold resistance, salt and alkali resistance and disease resistance. After treatment with CCC, the number of stomata and transpiration rate of leaves decrease, which can enhance drought resistance. It can be seen that CCC not only has the ability to promote plant growth, but also has the ability to delay cell elongation. Therefore, when using CCC, it is necessary to find the most suitable dosage and give full play to its function of regulating plant growth. If the dosage is inappropriate or the formula with other components is inappropriate, it is impossible to predict what function CCC will play. It's found in this invention that adding specific amounts of mannitol, ABA, CCC and sucrose to the MS medium can well preserve the Hosta plantagineain vitro plantlets, extend storage period to 6 months with a 100% survival rate. After the reculture, the phenotype of plants is stable without degeneration or variation. Even if the MS medium added with mannitol, abscisic acid, CCC and sucrose is adopted, if the dosage of the present application is not adopted, a better preservative effect cannot be realized. Therefore, this invention provides the application of the medium of this invention in preserving the Hosta plantaginea in vitro plantlets and in delaying the growth speed of the Hostaplantagineain vitro plantlets. On the other hand, this invention provides an application of the preservative medium of Hosta plantaginea in vitro plantlets, specifically comprising culturing Hosta plantagineain vitro plantlets in the preservative medium. The Hosta plantaginea in vitro plantlets are in vitro plantlets with strong and consistent growth. The Hosta plantaginea in vitro plantlets are obtained via propagation with the following methods: sterile Hosta plantaginea in vitro plantlets are cultured in the medium of MS+3% sucrose +0. 7 % agar (pH 5.8) and preserved for 30-45 days at the culture temperature of 20-25 °C , illumination intensity of 2000-30001x and illumination duration of 12-14h/d. In the preservative process, this invention has the preservation conditions of culture temperature of 20-25 °C , illumination intensity of 2000-30001x and illumination duration of 12-14h/d. Preferably, the preservation conditions are culture temperature of 25C, illumination intensity of 20001x and illumination duration of 14h/d.
The beneficial effects of this invention are: Through a large number of studies, with this invention, it is found that sucrose, mannitol, ABA and CCC are added into the MS medium according to a certain dosage ratio can be used for preserving the Hosta plantaginea in vitro plantlets for 6 months. The preservation method has the advantages of simple operation and low cost, and overcomes the defects of the preservation method reported in the prior art that the preservation method can be stored for at most 3 months, the preservation time is short, and the plant variation and degeneration are caused by frequent subculture. The method of the invention avoids the loss or destruction of germplasm caused by the influence of natural disasters in the field nursery preservation, also reduces the subculture frequency, greatly lowers the preservation cost of the Hosta plantagineain vitro plantlets, and is a safe, reliable, simple and effective method for preserving the germplasm resources of Hosta plantaginea. Applying the method of the invention to the actual production will obtain good economic benefits.
BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 shows the growth state of the controlled Hosta plantaginea in vitro plantlets preserved in Embodiment 3, where a is the growth state after one month of preservation; b is the growth state after 2 months of preservation; and c is the growth state after 3 months of preservation. Fig. 2 shows the growth state of the Hosta plantaginea in vitro plantlets preserved with delayed growth in Embodiment 3 (1), where a is the growth state after 2 months of preservation; b is the growth state after 4 months of preservation; and c is the growth state after 6 months of preservation. Fig. 3 shows the growth state of the Hosta plantaginea in vitro plantlets preserved with delayed growth in Embodiment 3 (2), where a is the growth state after one month of preservation; b is the growth state after two months of preservation; c is the growth state after three months of preservation; d is the growth state after four months of preservation; e is the growth state after five months of preservation; and f is the growth state after 6 months of preservation. Fig. 4 shows the growth state of the Hosta plantaginea in vitro plantlets after 1 month of reculture after preservation with delayed growth for 6 months in Embodiment 4 (1).
Fig. 5 shows the growth state of the Hosta plantaginea in vitro plantlets after 1 month of reculture after preservation with delayed growth for 6 months in Embodiment 4 (2).
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The following embodiments serve to illustrate this invention, and are not intended to limit its scope. Unless otherwise specified, the technical means used in the embodiments are conventional and well-known to those skilled in the field, and all the raw materials used are commercially available. Embodiment 1 Preservation Method of Hosta PlantagineaIn Vitro Plantlets Experimental Materials: Test-Tube Plantlets from National Crop Genebank of China The specific experimental steps are as follows: (1) The growth method of in vitro plantlets after propagation is as follows: the sterile Hosta plantagineain vitro plantlets are inoculated into the medium of MS+3% sucrose+0.7% agar (pH5.8) for 30-45 days at the culture temperature of 20-25 °C
, illumination intensity of 2000-30001x and illumination duration of 12-14h/d. (2) The method of delayed growth preservation is as follows: the sterile Hosta plantagineaplantlets with four leaves and one heart (5±0.5cm in height), which grow healthily and uniformly in step (1), are inoculated into the delayed growth preservation medium of MS+50g/L sucrose +20g/L mannitol +1.0mg/L ABA (ABA)+20mg/L CCC +0.7% agar (pH5.8) for 6 months at the culture temperature of -25°C, illumination intensity of 2000-30001x and illumination duration of 12-14h/d;
or Select the sterile Hosta plantaginea plantlets with four leaves and one heart (5±0.5cm in height), which grow healthily and uniformly in step (1), are inoculated into the delayed growth preservation medium of MS+50g/L sucrose +20g/L mannitol +1.0mg/L ABA+40mg/L CCC +0.7% agar (pH5.8) for 6 months at the culture temperature of 20-25 °C , illumination intensity of 2000-30001x and illumination
duration of 12-14h/d. (3) The method of reculture is as follows: the materials preserved in step (2) are inoculated into the reculture medium of MS+3% sucrose+0.7% agar (pH5.8) for ~60 days at the culture temperature of 20-25 °C , illumination intensity of 2000
30001x and illumination duration of 12-14h/d.
Embodiment 2 Survival Rate of Hosta PlantagineaIn Vitro Plantlets Preserved for 6 Months Sterile Hosta plantagineaplantlets (4 leaves and 1 heart, 5±0.5cm in height), which grow healthily and uniformly after 30~45 days in the medium of MS+3% sucrose+0.7% agar (pH5.8), are inoculated into the 11 preservation media, with the same culture conditions as Embodiment 1. It aims to study the effect of different preservation media on survival rate. Embodiment 3 Height Change of Hosta Plantaginea In Vitro Plantlets in Delayed Growth Preservation (1) Sterile Hosta plantagineaplantlets (4 leaves and 1 heart, 5±0.5cm in height), which grow healthily and uniformly after 30~45 days in the medium of MS+3% sucrose+0.7% agar (pH5.8) are inoculated into the control medium (MS+3% sucrose+0.7% agar, pH5.8) and delayed growth preservation medium (MS+50g/L sucrose +20g/L mannitol +1.Omg/L ABA +20mg/L CCC+0.7% agar, pH5.8), namely the medium in Scheme A of Embodiment 2. The culture conditions are the same as those in Embodiment 1. It aims to study the effect of medium in Scheme A of Embodiment 2 on the height of surviving plants. (2) Sterile Hosta plantagineaplantlets (4 leaves and 1 heart, 5±0.5cm in height), which grow healthily and uniformly after 30~45 days in the medium of MS+3% sucrose+0.7% agar (pH5.8), are inoculated into the control medium (MS+3% sucrose+0.7% agar, pH5.8) and delayed growth medium (MS+50g/L sucrose +20g/L mannitol +1.Omg/L ABA +40mg/L CCC +0.7% agar, pH5.8), namely the medium in Scheme E of Embodiment 2. The culture conditions are the same as those in Embodiment 1. It aims to study the effect of delayed growth medium on the height of surviving plants. Embodiment 4 Heights and Phenotype Changes of Hosta PlantagineaIn Vitro Plantlets after Reculture in Delayed Growth Preservation (1) Sterile Hosta plantagineaplantlets (4 leaves and 1 heart, 5±0.5cm in height), which grow healthily and uniformly after 30~45 days in the medium supplemented with MS+3% sucrose+0.7% agar (pH5.8), are inoculated into the control medium (MS+3% sucrose+0.7% agar, pH5.8) for 60 days and the medium in Scheme A of Embodiment 2 with MS+50g/L sucrose +20g/L mannitol +1.mg/LABA +20mg/L CCC+0.7% agar (pH5.8) for 6 months. Next, the reculture culture will be performed
in aforesaid plantlets for 60 days before the phenotypic examination. The culture conditions are the same as those in Embodiment 1. It aims to study the effects of different preservation media on phenotypic indexes of reculture plants, such as plant height, divergence, compactness, roundness and relative chlorophyll content. (2) Sterile Hosta plantagineaplantlets (4 leaves and 1 heart, 5±0.5cm in height), which grow healthily and uniformly after 30~45 days in the medium supplemented with MS+3% sucrose+0.7% agar (pH5.8) are inoculated into the control medium (MS+3% sucrose+0.7% agar, pH5.8) for 60 days and the slow growth preservation medium supplemented with MS+50g/L sucrose +20g/L mannitol +1.Omg/L ABA +40mg/L CCC +0.7% agar (pH5.8) for 6 months. Next, the reculture will be performed in aforesaid plantlets for 60 days before the phenotypic examination. The culture conditions are the same as those in Embodiment 1. It aims to study the effects of different preservation media on phenotypic indexes and senescence indexes of reculture plants, such as plant height, divergence, compactness, roundness and relative chlorophyll content. Scheme; Plant Height (cm); Divergence; Compactness; Control; 6.65±0.400; 1161.7±122.5; 0.405±0.029; Slow Growth Preservation Scheme; 6.40±0.410; 1247.3±53.5; 0.490±0.065; Scheme; Roundness; Relative Chlorophyll Content (%); Control; 256.9±37.56; 96.63±1.806; Slow Growth Preservation Scheme; 221.1±21.45; 97.33±0.664
Table 5 Changes in Height and Phenotype of Hosta Plantaginea In Vitro Plantlets after Reculture in Delayed Growth Preservation
Scheme Plant Height (cm) Divergence Compactness Control 6.65±0.400 1161.7±122.5 0.405±0.029 Slow Growth Preservation Scheme 6.40±0.410 1247.3±53.5 0.490±0.065 Roundness Relative Scheme Chlorophyll Content ( % )
Control 256.9±37.56 96.63±1.806 Slow Growth Preservation Scheme 221.1±21.45 97.33±0.664
Although this invention has been described in detail through general description, specific embodiments and experiments, it is obvious to those skilled in this field that some modifications or improvements may be made on this basis. Therefore, such
modifications or improvements made without deviating from the spirit of this invention should fall within the scope of protection required by the invention.
Claims (9)
1. A preservation medium of Hosta plantaginea in vitro plantlets, characterized in that the said medium is supplemented with MS+(502.5)g/L sucrose+(20±1)g/L mannitol+(1.0±0.05)mg/L ABA+19-42mg/L CCC+0.7% agar (pH 5.8).
2. A preservation medium of Hosta plantaginea in vitro plantlets as stated in Claim 1, characterized in that the said medium is supplemented with MS+(50±2.0)g/L sucrose+(20±0.5)g/L mannitol+(1.0±0.04)mg/L ABA+20-40mg/L CCC+0.7% agar.
3. A preservation medium of Hosta plantaginea in vitro plantlets as stated in Claim 1, characterized in that the said medium is supplemented with MS+(50±1.0)g/L sucrose+(20±0.2)g/L mannitol+(1.0±0.02)mg/L ABA+20-40mg/L CCC+0.7% agar (pH5.8).
4. A preservation medium of Hosta plantaginea in vitro plantlets as stated in Claim 1, characterized in that the said medium is supplemented with MS+50g/L sucrose+20g/L mannitol+1.Omg/L ABA+20mg/L CCC or 40 mg/L CCC +0.7% agar (pH5.8).
5. The application of the medium according to any of Claims 1-4 in preserving Hosta plantaginea in vitro plantlets or slowing the growth rate of Hosta plantaginea in vitro plantlets.
6. The application as stated in Claim 5, characterized in that the Hosta plantagineain vitro plantlets are cultured in the preservation medium.
7. The application as stated in Claim 5, characterized in that the said Hosta plantaginea in vitro plantlets are in vitro plantlets with strong growth and consistent growth.
8. The application as stated in Claim 5, characterized in that the said Hosta plantaginea in vitro plantlets are obtained through propagation by the following method: culture sterile Hosta plantaginea in vitro plantlets in the medium supplemented with MS+3 o sucrose+0.7% agar (pH5.8) and culture for 30~45 days,
at the culture temperature of 20 25°C, illumination intensity of 2000 30001x and
illumination duration of 12~14h/d
9. The application as stated in any of Claims 5-8, characterized in that the preservation conditions are culture temperature of 20-25 °C, illumination intensity of
2000-30001x and illumination duration of 12-14h/d.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2021107353A AU2021107353A4 (en) | 2021-08-25 | 2021-08-25 | A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2021107353A AU2021107353A4 (en) | 2021-08-25 | 2021-08-25 | A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AU2021107353A4 true AU2021107353A4 (en) | 2021-12-16 |
Family
ID=78948984
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU2021107353A Ceased AU2021107353A4 (en) | 2021-08-25 | 2021-08-25 | A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application |
Country Status (1)
| Country | Link |
|---|---|
| AU (1) | AU2021107353A4 (en) |
-
2021
- 2021-08-25 AU AU2021107353A patent/AU2021107353A4/en not_active Ceased
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wubs et al. | Abortion of reproductive organs in sweet pepper (Capsicum annuum L.): a review | |
| Xu et al. | Technique of grafting with Wufanshu (Vaccinium bracteatum Thunb.) and the effects on blueberry plant growth and development, fruit yield and quality | |
| Yang et al. | Regulated deficit irrigation: an effective way to solve the shortage of agricultural water for horticulture | |
| Pant et al. | Various culture techniques for the mass propagation of medicinal orchids from Nepal | |
| Nin et al. | Effects of environmental factors on seed germination and seedling establishment in bilberry (Vaccinium myrtillus L.) | |
| Dolce et al. | In vitro propagation and germplasm conservation of wild orchids from South America | |
| CN111837947B (en) | Culture medium for preserving test-tube plantlets of Hosta plantaginea and application of culture medium | |
| Tsegaw | Response of potato to paclobutrazol and manipulation of reproductive growth under tropical conditions | |
| Premalatha et al. | Plant training and spatial arrangement for yield improvements in greenhouse cucumber (Cucumis sativus L.) varieties | |
| Minhas | Potato: production strategies under abiotic stress | |
| AU2021107353A4 (en) | A Preservation Medium of Hosta Plantaginea In Vitro Plantlets and Its Application | |
| AU2021107357A4 (en) | A Preservation Method of Hosta Plantaginea In vitro Plantlets | |
| CN111802379B (en) | Method for storing test-tube plantlets of Hosta plantaginea | |
| Mendonça et al. | Accumulated leaf number in strawberry cultivars grown in a greenhouse | |
| Dahal | Post Harvest Handing of Cut-Flower Rose | |
| Jain et al. | Utility of plant growth regulators in improving fruit production | |
| Della Rahayu et al. | Asymbiotic seed germination and plantlet development of Dendrobium spectabile (Blume) Miq. | |
| Rao | Cultivation, economics and marketing of Phyllanthus species | |
| Nan | Effects of pre-and postharvest calcium supplementation on longevity of sunflower (Helianthus annuus cv. Superior Sunset) | |
| Abdel | Water relation in lettuce (Lactuca sativa L. var. Longifolia) | |
| Kumar et al. | Studies on Influence of Growth Retardants on Flower Crops | |
| Tazeem et al. | The impact of climate change on vegetable production | |
| CN113854058B (en) | Method for improving drought resistance and salt resistance of fringed pink | |
| Cavalcante | Mango Flowering: Factors Involved in the Natural Environment and Associated Management Techniques for Commercial Crops | |
| WO2021172594A1 (en) | Plant heat resistance or drought resistance improving agent, salt tolerance improving agent, activity improving agent |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FGI | Letters patent sealed or granted (innovation patent) | ||
| MK22 | Patent ceased section 143a(d), or expired - non payment of renewal fee or expiry |