CN117016388A - Application of multiwall carbon nanotubes in promoting growth of gametophyte and/or sporophyte production of pteridophyte - Google Patents
Application of multiwall carbon nanotubes in promoting growth of gametophyte and/or sporophyte production of pteridophyte Download PDFInfo
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- CN117016388A CN117016388A CN202310681066.0A CN202310681066A CN117016388A CN 117016388 A CN117016388 A CN 117016388A CN 202310681066 A CN202310681066 A CN 202310681066A CN 117016388 A CN117016388 A CN 117016388A
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- 239000002048 multi walled nanotube Substances 0.000 title claims abstract description 43
- 230000001737 promoting effect Effects 0.000 title claims abstract description 22
- 238000004519 manufacturing process Methods 0.000 title claims description 19
- 238000011161 development Methods 0.000 claims abstract description 25
- 241000874833 Salvinia minima Species 0.000 claims abstract description 21
- 239000001963 growth medium Substances 0.000 claims description 26
- 238000012258 culturing Methods 0.000 claims description 13
- 239000002609 medium Substances 0.000 claims description 13
- 239000012154 double-distilled water Substances 0.000 claims description 12
- 239000000725 suspension Substances 0.000 claims description 12
- 229920001817 Agar Polymers 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 241000985694 Polypodiopsida Species 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 239000007640 basal medium Substances 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 4
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 4
- 230000018109 developmental process Effects 0.000 abstract description 22
- 230000000694 effects Effects 0.000 abstract description 9
- 230000035784 germination Effects 0.000 abstract description 9
- 210000004681 ovum Anatomy 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- 241000196324 Embryophyta Species 0.000 description 7
- 102000002322 Egg Proteins Human genes 0.000 description 6
- 108010000912 Egg Proteins Proteins 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 238000005286 illumination Methods 0.000 description 5
- 230000004763 spore germination Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 241000736301 Pteridaceae Species 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 239000002086 nanomaterial Substances 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 230000014639 sexual reproduction Effects 0.000 description 3
- 240000005893 Pteridium aquilinum Species 0.000 description 2
- 235000009936 Pteridium aquilinum Nutrition 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 241001196073 Alsophila spinulosa Species 0.000 description 1
- 241001148499 Ceratopteris Species 0.000 description 1
- 241001261549 Salviniales Species 0.000 description 1
- 241000592342 Tracheophyta Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000006543 gametophyte development Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
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- 239000000203 mixture Substances 0.000 description 1
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- 150000002894 organic compounds Chemical class 0.000 description 1
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- 230000001568 sexual effect Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000003911 water pollution Methods 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
- A01H4/002—Culture media for tissue culture
Abstract
The invention provides an application of a multiwall carbon nanotube in promoting the development of gametophytes and/or the generation of sporophytes of pteridophytes, belonging to the technical field of pteridophyte culture. The invention provides the multi-wall carbon nano tube for the first time, which has the effect of promoting the development of the gametophyte and/or the generation of the sporophyte of the pteridophyte, and the adoption of the multi-wall carbon nano tube with proper concentration for treatment can obviously promote the development of the gametophyte and the generation of the sporophyte of the pteridophyte, and the number of sperms is obviously increased. The 0.5-2.5 mg/L multiwall carbon nanotube treatment can lead the germination of the water fern spores to be advanced by about 15d, wherein the 0.5-1.0 mg/L multiwall carbon nanotube treatment has the most obvious promotion effect before the flaky stage, and the 2.5mg/L multiwall carbon nanotube treatment has more obvious promotion effect after the flaky stage.
Description
Technical Field
The invention belongs to the technical field of fern culture, and particularly relates to application of a multiwall carbon nanotube in promoting development of gametophytes and/or production of sporophytes of ferns.
Background
The evolution status of the pteridophyte is special, the pteridophyte is not only a higher spore plant, but also a lower vascular plant, and the pteridophyte has sporophyte and gametophyte for 2 generations, can independently survive, is unique in the plant kingdom, and is an important material for researching plant evolution and phylogenetic development. Chinese pteridophyte resources are rich, the Chinese pteridophyte resources are applied to various aspects such as eating, medicine, environmental indication and the like, and part of pteridophytes have important research values due to the self specificity. However, due to the problems of destruction of natural ecological environment, unreasonable development and utilization, short self-life of spores, difficult germination and the like, some precious and rare wild pteridophyte resources are extinct. The basic research of the related propagation technology is enhanced, and the method has important research value and practical significance for the utilization and protection of the germplasm resources of the pteridophytes.
The water fern is a plant of the genus Pteridaceae (Pteridaceae) genus Pteridaceae (Ceratopteris brongn.). The water fern is considered as an important nitrogen-fixing plant and plays an important role in improving soil quality, crop yield and the like. Water ferns are also used as fertilizers or feed in some areas. The water fern can absorb and degrade pollutants such as heavy metal ions, organic compounds and the like, so that the water fern is widely applied to the treatment of water pollution of lakes, reservoirs and the like. Meanwhile, the ecological and environmental adaptability of the water fern can be researched, and a reference can be provided for environmental treatment. Moreover, the water fern is an original plant and has important significance for researching biological evolution and earth life history. Meanwhile, the water fern is an important constituent part of the wetland ecosystem, and has important significance for wetland protection and biodiversity maintenance. In conclusion, the water fern has important economic, ecological and scientific values, strengthens the basic research of the propagation technology related to the water fern, and has important research value and practical significance for the utilization and protection of the germplasm resources of ferns.
The multi-wall carbon nano tube is taken as a one-dimensional nano material, has small radial dimension, extremely high strength and extremely high toughness, and simultaneously has excellent electrical and chemical properties, and is considered as a future super fiber. In recent years, with the deep research of multi-wall carbon nanotubes and nano materials, the wide application prospect of the nano materials is also continuously shown. Conventionally, there have been few applications of multiwall carbon nanotubes in ferns, and the studies of promoting the germination of spores of Alsophila spinulosa (wall. Ex hook.) have been disclosed to achieve only the effect of promoting the formation of filament stages, but for the development of ferns gametophytes, it has been proposed to promote spore germination through several processes such as filament stages, lamellar stages, protoleaf stages and sporophyte formation, but for the whole gametophytes of ferns, the spore germination is implied as long as the pseudoroot extends beyond the spore wall, so that the prior art scheme can finally form lamellar bodies, protoleaf bodies and sporophytes, and it has not been known from knowledge, and whether there is a difference in the effect of adding multiwall carbon nanotubes of different concentrations on each stage of gametophyte development. It can be seen that the prior art is not yet in depth and that there is no report about whether the addition of multiwall carbon nanotubes has an effect on the formation of sexual organs (cervical ova and sperm apparatus).
Disclosure of Invention
In view of the above, the present invention aims to provide an application of multi-walled carbon nanotubes in promoting the development of gametophytes and/or the production of sporophytes of pteridophytes, and the present invention provides a multi-walled carbon nanotube with a proper concentration for the first time, which can obviously promote the development of gametophytes and the production of sporophytes of pteridophytes, and the number of spermatids is obviously increased.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an application of multi-wall carbon nano-tubes in promoting the development of gametophytes and/or the production of sporophytes of pteridophytes.
Preferably, the fern plant comprises a water fern.
The invention also provides a culture medium for promoting the development of the gametophyte and/or the production of the sporophyte of the pteridophyte, which comprises a basic culture medium and 0.1-5.0 mg/L multiwall carbon nano tube.
Preferably, the basal medium comprises 1/2MS medium.
Preferably, the basal medium consists of 1/2MS medium and 7.5g/L agar.
The invention also provides application of the culture medium in the culture of pteridophytes.
The invention also provides a method for promoting the development of gametophytes and/or the production of sporophytes of pteridophytes, which comprises the following steps: mixing the sterilized pteridophyte spores with double distilled water to obtain spore suspension; inoculating the spore suspension into the culture medium of any one of the above.
Preferably, the sterilization comprises the steps of: the pteridophyte spores are treated with NaClO with the volume fraction of 3% 3 Sterilizing for 2min, and cleaning with double distilled water.
Preferably, the culture conditions are 25 ℃, the illumination time is 18h/d, and the illumination intensity is 100-300 mu mol/(m) 2 ·s -1 )。
The invention has the beneficial effects that:
the invention provides the multi-wall carbon nano tube for the first time, which has the effect of promoting the development of the gametophyte and/or the generation of the sporophyte of the pteridophyte, and the adoption of the multi-wall carbon nano tube with proper concentration for treatment can obviously promote the development of the gametophyte and the generation of the sporophyte of the pteridophyte, and the number of sperms is obviously increased. The 0.5-2.5 mg/L multiwall carbon nanotube treatment can lead the germination of the water fern spores to be advanced by about 15d, wherein the 0.5-1.0 mg/L multiwall carbon nanotube treatment has the most obvious promotion effect before the flaky stage, and the 2.5mg/L multiwall carbon nanotube treatment has more obvious promotion effect after the flaky stage.
Drawings
FIG. 1 is a graph showing the results of the development of the gametophytes of the water fern of comparative example 1 and examples 5 to 8, wherein A1, A2, A3, A4, A5 are germination conditions of the water fern spore cultures 15d, 19d, 32d, 41d of comparative example 1, respectively; b1, B2, B3, B4 and B5 are germination conditions of the culture of the water fern spores of example 5 (0.5 mg/L multiwall carbon nanotube treatment group) for 7d, 15d, 19d, 32d and 32d, respectively; c1, C2, C3, C4, C5 are germination of the culture of the Pteridium aquilinum spores of example 6 (1.0 mg/L multiwall carbon nanotube treated group) for 7d, 15d, 19d, 32d, 41d, respectively; d1, D2, D3, D4, D5, D6 are germination conditions for the culture of the water fern spores 7D, 12D, 15D, 19D, 32D, 41D, respectively, of example 7 (2.5 mg/L multiwall carbon nanotube treated group); e1, E2, E3, E4, E5, E6 are germination of the culture of Pteridium aquilinum spores 9d, 16d, 18d, 28d, respectively, of example 8 (5.0 mg/L multiwall carbon nanotube treated group).
Detailed Description
The invention provides an application of multi-wall carbon nano-tubes in promoting the development of gametophytes and/or the production of sporophytes of pteridophytes.
The specific source of the multiwall carbon nanotubes is not particularly limited, and any commercially available product conventionally used in the art can be used. The specific source of the pteridophyte is not particularly limited in the present invention, and in the present invention, the pteridophyte preferably includes a pteridophyte.
The invention also provides a culture medium for promoting the development of the gametophyte and/or the production of the sporophyte of the pteridophyte, which comprises a basic culture medium and 0.1-5.0 mg/L multiwall carbon nano tube.
In the present invention, the basal medium is a medium used for culturing ferns, preferably comprising 1/2MS medium. The specific composition of the 1/2MS culture medium is not particularly limited, and the 1/2MS culture medium which is conventional in the art can be adopted. In the present invention, the basal medium is more preferably composed of 1/2MS medium and 7.5g/L agar. The specific source of the agar is not particularly limited, and the agar can be obtained by using products conventionally and commercially available in the art. In the culture medium of the present invention, the concentration of the multiwall carbon nanotubes is preferably 0.5 to 2.5mg/L.
The invention also provides application of the culture medium in the culture of pteridophytes.
The invention also provides a method for promoting the development of gametophytes and/or the production of sporophytes of pteridophytes, which comprises the following steps: mixing the sterilized pteridophyte spores with double distilled water to obtain spore suspension; inoculating the spore suspension into the culture medium of any one of the above.
In the present invention, the water is preferably double distilled water, and the sterilization preferably includes the steps of: the pteridophyte spores are treated with NaClO with the volume fraction of 3% 3 Sterilizing for 2min, and cleaning with double distilled water. The number of times of sterilization is preferably 1, and after sterilization, centrifugation is carried out, wherein the conditions of the centrifugation are preferably 12000r/minHeart for 2min. The washing times are preferably 3 times, the washing is preferably centrifugal washing, the centrifugal washing conditions are preferably 12000r/min for 2min, after one centrifugal washing is finished, the supernatant is poured out, and the settled spores are added with double distilled water for centrifugal washing. After the cleaning is finished, double-distilled water is preferably adopted for overnight use. The invention relates to NaClO 3 And the specific source of double distilled water are not particularly limited. In the present invention, the culture conditions are preferably 25℃and the illumination time is 18h/d, and the illumination intensity is 100 to 300. Mu. Mol/(m) 2 ·s -1 ) The illumination intensity is more preferably 150 to 250. Mu. Mol/(m) 2 ·s -1 )。
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
In the following examples, conventional methods are used unless otherwise specified.
Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1
A culture medium for promoting the development of gametophyte and/or the production of sporophyte of pteridophyte comprises 1/2MS culture medium, 7.5g/L agar and 0.5mg/L multiwall carbon nanotube.
Example 2
A culture medium for promoting the development of gametophyte and/or the production of sporophyte of pteridophyte comprises 1/2MS culture medium, 7.5g/L agar and 1.0mg/L multiwall carbon nanotube.
Example 3
A culture medium for promoting the development of gametophyte and/or the production of sporophyte of pteridophyte comprises 1/2MS culture medium, 7.5g/L agar and 2.5mg/L multiwall carbon nanotubes.
Example 4
A culture medium for promoting the development of gametophyte and/or the production of sporophyte of pteridophyte comprises 1/2MS culture medium, 7.5g/L agar and 5.0mg/L multiwall carbon nanotubes.
Example 5
Taking the pteridophyte as an explant, throughNaClO with 3% volume fraction 3 Sterilizing for 1 time, centrifuging for 2min at 12000r/min, washing with double distilled water for 3 times (washing condition is 12000r/min for 2 min), and soaking with double distilled water overnight to obtain herba Pteridis Multifidae spore suspension for use.
Inoculating the suspension of herba Pteridis Multifidae spore into the culture medium of example 1, and placing at 25deg.C for 18h/d under light with light intensity of 150 μmol/(m) 2 ·s -1 ) Culturing under the condition, observing spore germination every day, and recording by using a microscope for photographing.
The results are shown in FIGS. 1B1 to B5: after 7d of culture, spores in the 0.5mg/L multiwall carbon nanotube treated group were observed to have developed into a sheet (B1); after 15d of culture, sperm cell (B2) was produced on the pellet; culturing for 19 days, and then developing the flaky body into a protoleaf body, wherein a cervical ovum device is arranged on the protoleaf body to generate (B3); culturing until 32d, increasing the number of cervical ova (B4), and completing sexual reproduction on part of the protozoon to generate sporophytes (B5), and entering sporophyte generation.
Example 6
The difference from example 5 is that the suspension of the water fern spores was inoculated into the medium of example 2 for cultivation, and the rest was the same as example 5.
The results are shown in FIGS. 1C1 to C5: after 7d of culture, spores in the 1.0mg/L multiwall carbon nanotube treated group were observed to have developed into a sheet (C1); after 15d of culture, sperm cell (C2) was produced on the pellet; culturing for 19d, and then developing the flaky body into a protoleaf body, wherein a cervical ovum device is arranged on the protoleaf body to generate (C3); culturing for 32 days, increasing the number of cervical ovum devices, and generating nutrition growth of partial protophylls near the growth point to generate small flaky bodies (C4); culturing to 41d to generate sporophyte on the protophyll, and entering sporophyte generation (C5).
Example 7
The difference from example 5 is that the suspension of the water fern spores was inoculated into the medium of example 3 for cultivation, and the rest was the same as example 5.
The results are shown in FIGS. 1D1 to D6: spore germination was observed in the 2.5mg/L multiwall carbon nanotube treated group after 7D of culture, yielding a filament (D1) consisting of several cells; after 12D of culture, the filaments develop into sheets, and a large number of sperm cells (D2) are produced on the sheets; after 15D of culture, the lamina develops into a protoleaf (D3); after 19D of culture, the primordial leaf body is provided with a cervical ovum device, and the volume is larger (D4); culturing for 32 days, increasing the number of cervical ovum devices, and generating nutrition growth of partial protophylls near the growth point to generate small flaky bodies (D5); culturing to 41D to generate sporophyte on the protophyll, and entering sporophyte generation (D6).
Example 8
The difference from example 5 is that the suspension of the water fern spores was inoculated into the medium of example 4 for cultivation, and the rest was the same as in example 5.
The results are shown in FIGS. 1E1 to E5: after 9d of culture, a small number of spores in the 5.0mg/L multiwall carbon nanotube treated group were observed to have pseudoroots protruding out of the spore wall (E1); after 16d of cultivation, a pellet (E2) was produced; after 18d of culture, the tablet body is provided with a spermatid (E3); culturing to 28d, wherein most gametophytes are still in a lamellar body stage (E4), and sexual reproduction production sporophytes are completed on most of the gametophytes (E5), and the gametophytes enter sporophyte generation.
Comparative example 1
The difference from example 5 is that the suspension of the water fern spores is inoculated into the following medium for cultivation, the rest is the same as example 5: the culture medium consisted of 1/2MS medium and 7.5g/L agar, and no multiwall carbon nanotubes were added. The results are shown in FIGS. 1A1 to A5: after 15d of culture, it was observed that most of spores in the 0.0mg/L multiwall carbon nanotube treated group had not germinated (A1), only a small amount of spores had cracked outer walls, or that a few cells and pseudoroots had protruded out of spore walls (A2); culturing for 19d to obtain a sheet with sperm cell (A3); after 32d of culture, the flaky body develops into a protoleaf body, and a cervical ovum device is arranged on the protoleaf body to generate (A4); culturing to 41d, wherein sexual reproduction has been completed on part of the protophyll to produce sporophytes (A5), and entering sporophyte generation.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (9)
1. Use of multiwall carbon nanotubes for promoting the development of gametophytes and/or the production of sporophytes of ferns.
2. The use according to claim 1, wherein said fern comprises water fern.
3. A culture medium for promoting the development of gametophytes and/or the production of sporophytes of pteridophytes is characterized by comprising a basic culture medium and 0.1-5.0 mg/L multi-wall carbon nano-tubes.
4. A culture medium according to claim 3, wherein the basal medium comprises 1/2MS medium.
5. A culture medium according to claim 3, characterized in that the basal medium consists of 1/2MS medium and 7.5g/L agar.
6. Use of the medium according to any one of claims 3 to 5 for the cultivation of ferns.
7. A method for promoting the development of gametophytes and/or the production of sporophytes in a pteridophyte comprising the steps of: mixing the sterilized pteridophyte spores with double distilled water to obtain spore suspension; inoculating the spore suspension into the culture medium according to any one of claims 3 to 5 for culture.
8. The method of claim 7, wherein the sterilizing comprises the steps of: the pteridophyte spores are treated with NaClO with the volume fraction of 3% 3 Sterilizing for 2min, and cleaning with double distilled water.
9. The method according to claim 8, wherein the culturing is carried out at 25℃for 18 hours/d under light with an intensity of 100 to 300. Mu. Mol/(m) 2 ·s -1 )。
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| CN117016388B (en) | 2024-04-19 |
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