CN108605988B - Irritant for plant grafting propagation and preparation method thereof - Google Patents
Irritant for plant grafting propagation and preparation method thereof Download PDFInfo
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- CN108605988B CN108605988B CN201810334846.7A CN201810334846A CN108605988B CN 108605988 B CN108605988 B CN 108605988B CN 201810334846 A CN201810334846 A CN 201810334846A CN 108605988 B CN108605988 B CN 108605988B
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- 238000002360 preparation method Methods 0.000 title description 7
- 239000002085 irritant Substances 0.000 title description 3
- 231100000021 irritant Toxicity 0.000 title description 3
- 239000001963 growth medium Substances 0.000 claims description 30
- 241000223221 Fusarium oxysporum Species 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 22
- 238000012258 culturing Methods 0.000 claims description 20
- 241000222290 Cladosporium Species 0.000 claims description 15
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- 238000011218 seed culture Methods 0.000 claims description 10
- OTCCIMWXFLJLIA-BYPYZUCNSA-N N-acetyl-L-aspartic acid Chemical compound CC(=O)N[C@H](C(O)=O)CC(O)=O OTCCIMWXFLJLIA-BYPYZUCNSA-N 0.000 claims description 8
- 241001149955 Cladosporium cladosporioides Species 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 5
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 5
- 239000011790 ferrous sulphate Substances 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 238000011081 inoculation Methods 0.000 claims description 5
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 5
- 238000009630 liquid culture Methods 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 5
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 230000004083 survival effect Effects 0.000 abstract description 21
- 230000008635 plant growth Effects 0.000 abstract description 18
- 244000005700 microbiome Species 0.000 abstract description 11
- 241000233866 Fungi Species 0.000 abstract description 8
- 102000018997 Growth Hormone Human genes 0.000 abstract description 7
- 108010051696 Growth Hormone Proteins 0.000 abstract description 7
- 239000000122 growth hormone Substances 0.000 abstract description 7
- 239000003375 plant hormone Substances 0.000 abstract description 6
- 230000002195 synergetic effect Effects 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 38
- 230000012010 growth Effects 0.000 description 18
- 230000000694 effects Effects 0.000 description 8
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- 206010020649 Hyperkeratosis Diseases 0.000 description 7
- 241000207199 Citrus Species 0.000 description 6
- 235000020971 citrus fruits Nutrition 0.000 description 6
- 240000001548 Camellia japonica Species 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 4
- 235000018597 common camellia Nutrition 0.000 description 4
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 description 4
- 239000004062 cytokinin Substances 0.000 description 4
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- 229930192334 Auxin Natural products 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
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- 229930191978 Gibberellin Natural products 0.000 description 3
- 239000002363 auxin Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
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- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 2
- 241000223218 Fusarium Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
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- 230000008901 benefit Effects 0.000 description 2
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- 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 2
- 230000006872 improvement Effects 0.000 description 2
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
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- 229930195732 phytohormone Natural products 0.000 description 2
- 239000000021 stimulant Substances 0.000 description 2
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 2
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- 241000195493 Cryptophyta Species 0.000 description 1
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- 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 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000005712 elicitor Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical class C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 1
- 229960001669 kinetin Drugs 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/10—Aromatic or araliphatic carboxylic acids, or thio analogues thereof; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/12—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing the group, wherein Cn means a carbon skeleton not containing a ring; Thio analogues thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Environmental Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of plant grafting propagation, and discloses a stimulator for plant grafting propagation, which comprises fungi and plant hormones. The novel stimulant is prepared by combining traditional plant growth hormone and microorganisms, is reasonable in formula, is mutually synergistic, is convenient to use, and can improve the survival rate of scions in grafting.
Description
Technical Field
The invention belongs to the technical field of plant grafting propagation, and particularly relates to a stimulant for plant grafting propagation and a preparation method thereof.
Background
The grafting refers to a technology for transferring buds and branches of excellent plant varieties to stems and roots of another plant body and enabling the buds and the branches to heal up to form new independent plants, and is a widely applied technology in the seedling raising and production processes of horticultural crops. The biological principle of grafting survival is that a layer of brown membrane is formed by the residue of dead cells on the cut surface of the stock and the scion, then the cells around the wound are divided under the action of callus, cambium cells are activated, the membrane is broken to form callus, the callus cells are further differentiated to form new xylem inwards and new phloem outwards, and the xylem conduit of the stock and the phloem screen pipe are communicated to connect the conducting tissues. As new external plug cells are formed and connected with both plug cells, the new plug cells are finally healed to form a new plant.
A plant growth stimulant is an exogenous non-nutritive chemical that is usually delivered to the site of action in the plant and at lower concentrations promotes or inhibits certain links in the plant's life process, leading to a desire to meet human needs. The stimulators used in grafting generally include various plant hormones such as cytokinin, kinetin, and various hormones. Before grafting, the stock and the scion are generally soaked in a plant growth stimulant and are used for improving the activity of the callus, promoting the growth of the callus, promoting the germination of the scion, improving the chlorophyll content of the scion, enhancing photosynthesis, accelerating growth and other functions.
At present, the number of plants related to microecology studied is about hundreds, and the plants mainly comprise needle-leaved trees, algae, herbs, shrubs and other various species. As can be seen by analyzing the results of the current research, the endophytes of plants mainly include bacteria and fungi. The distribution range of the endophyte of the plant is very wide, and the endophyte is mainly in organs such as roots, stems, leaves, flowers, seeds, fruits and the like of the plant. In general, endophyte hyphae are present in lesser amounts in the leaves and heels of the plant, but are present in the largest amounts in the seeds and leaf sheaths. The endophytes of the plants are interacted and mutually influenced, so that a good ecological balance is established.
When the healthy plants have plant endophytes, some symptoms cannot be shown, however, when the autoimmunity of the plants is reduced, some endophytes are changed into pathogenic bacteria, and certain damage is caused to the plants. Endophytes are capable of producing certain metabolites that affect plant growth to some extent. From another perspective, the plant may provide conditions relevant for the production of the endophyte. Firstly, the nitrogen fixation function of the endophyte, part of the endophyte can survive under the condition of high sugar, and simultaneously, the nitrogen fixation function can be fully formed, so that the endophyte not only has obvious host specificity, but also has stronger acid resistance. Second, endophytes have some plant growth promoting effect because they can produce a plant growth promoting substance, such as gibberellins, auxins, cytokinins, etc. In addition, the endophyte can improve the absorption of nutrient elements such as P, N by plants, and simultaneously can compete with pathogenic bacteria for nutrition and space, thereby promoting the growth of plants. Third, adult plants are difficult to disinfect because they are bacterial and fungal in their growth, which makes tissue culture and genetic transformation difficult.
At present, the effect on plant ecological bacteria cannot be summarized, and partial documents are reported in the prior art. The influence of a fungal elicitor on the growth of a dendrobium officinale tissue culture, which is discovered in 2004 of the traditional Chinese medicine and pharmacology, that different bacterium types have different influence effects on the dendrobium officinale, and some strains can have obvious promotion effects on the dendrobium officinale, which are mainly shown in the aspects of biomass growth, plant growth, adventitious bud proliferation and root differentiation; some strains inhibit root differentiation. The survival rate of stem tip grafting of citrus seedlings can reach over 90 percent in 2008 of the university journal of Hunan agriculture (Nature science edition), the survival rate of stem tip grafting of citrus scion buds in an adult state of citrus is very low and is about 10 percent, and through preliminary analysis, the survival rate is probably related to the fact that the more citrus endophytes are carried by the scion buds in the adult state of citrus, different endophytes exist in citrus, the frequency of appearance in different organs is different, and the endophytes interfere with the normal growth of the scion.
Disclosure of Invention
Aiming at the technical problems in the prior art, the invention provides the stimulator for the grafting propagation of the plants and the preparation method thereof, the stimulator comprises phytohormone and fungi, is reasonable in compatibility, can improve the grafting survival rate and the growth speed, and promotes the formation of large crowns in a short time.
The invention is realized by the following technical scheme:
a stimulant for plant graft propagation comprising a fungus.
Further, the air conditioner is provided with a fan,
the fungi comprise Fusarium pachyspora or/and Cladosporium cladonioides.
Further, the air conditioner is provided with a fan,
the stimulant is prepared by the following process:
step 1) inoculating fusarium oxysporum on a PDA culture medium for culturing by streaking to obtain a single colony; selecting a single colony, inoculating the single colony on a seed culture medium for culture, and culturing at 30 ℃ for 24h to obtain fusarium oxysporum seed liquid for later use;
step 2) inoculating cladosporium dendritic on a PDA culture medium for culturing to obtain a single colony; selecting a single colony, inoculating the single colony to a PDA liquid culture medium for culture, and culturing at 30 ℃ for 24 hours to obtain a cladosporium dendritic seed solution for later use;
and 3) sequentially inoculating the fusarium oxysporum seed liquid and the cladosporium cladosporioides seed liquid into an MS culture medium, wherein the inoculation amounts are 3-5% and 4-7%, respectively, adding 6-BA, NAA and DA-6, and stirring at 100rpm for 3-6 hours at the temperature of 28-30 ℃ to obtain the fusarium oxysporum seed liquid.
Preferably, the first and second electrodes are formed of a metal,
the formula of the seed culture medium is as follows: 10g/L glucose, 6g/L yeast extract, 1g/L dipotassium phosphate, 1g/L potassium dihydrogen phosphate, 0.01g/L magnesium sulfate and 0.01g/L ferrous sulfate.
Preferably, the first and second electrodes are formed of a metal,
the fusarium oxysporum is ATCC 76222; the cladosporium cladosporioides is ATCC 34010.
Preferably, the first and second electrodes are formed of a metal,
the addition amount of the 6-BA is 50-100mg/L, the addition amount of the NAA is 40-70mg/L, and the addition amount of the DA-6 is 30-50 mg/L.
Compared with the prior art, the invention has the advantages that the following aspects are mainly included but not limited:
the novel stimulant is prepared by combining traditional plant growth hormone and microorganisms, has a reasonable formula and mutual synergy, is convenient to use, and can improve the activity of callus, promote the growth of the callus, promote the germination of scions and improve the survival rate of scions; through comparison tests, the specific microorganism is added into the stimulant to improve the scion survival rate, the growth amount of scion and the lateral bud drawing amount are both obviously improved, and the scion survival rate and the lateral bud drawing amount are the result of joint stimulation of the two microorganisms and the plant growth hormone.
The invention can establish a good ecological balance by selecting specific microorganisms which form symbiosis, interaction and mutual influence with plant tissues, generate substances for promoting plant growth, such as gibberellin, auxin, cytokinin and the like, improve the absorption of nutrient elements such as P, N and the like by plants, and simultaneously compete with pathogenic bacteria for nutrition and space, thereby promoting the growth of plants.
The stimulant of the invention realizes high survival rate of grafted plants, obviously improves the growth amount of scion and lateral bud shoot bud, has quick growth after survival of the grafted plants, can form large crown in a short period, has large crown width and more branches, and ensures that the plants are very robust, thereby improving the cost and economic benefit of camellia grafting.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the present application will be clearly and completely described below with reference to specific embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A stimulant for plant graft propagation, comprising a plant hormone and a fungus;
the preparation method specifically comprises the following steps:
inoculating fusarium oxysporum on a PDA culture medium for culturing by streaking to obtain a single colony; selecting a single colony, inoculating the single colony on a seed culture medium for culture, and culturing at 30 ℃ for 24h to obtain fusarium oxysporum seed liquid for later use; the formula of the seed culture medium is as follows: 10g/L glucose, 6g/L yeast extract, 1g/L dipotassium phosphate, 1g/L potassium dihydrogen phosphate, 0.01g/L magnesium sulfate and 0.01g/L ferrous sulfate;
inoculating cladosporium dendritic on a PDA culture medium for culturing to obtain a single colony; selecting a single colony, inoculating the single colony to a PDA liquid culture medium for culture, and culturing at 30 ℃ for 24 hours to obtain a cladosporium dendritic seed solution for later use;
sequentially inoculating fusarium oxysporum seed liquid and cladosporium dendritic seed liquid into an MS culture medium, wherein the inoculation amounts are 3% and 4%, respectively, adding 100mg/L of 6-BA, 70mg/L of NAA and 50mg/L of DA-6, and stirring at 100rpm for 5 hours at the temperature of 30 ℃ to obtain the fusarium oxysporum seed liquid.
The fusarium oxysporum is ATCC 76222; the cladosporium cladosporioides is ATCC 34010.
Example 2
A stimulant for plant graft propagation, comprising a plant hormone and a fungus;
the preparation method specifically comprises the following steps:
inoculating fusarium oxysporum on a PDA culture medium for culturing by streaking to obtain a single colony; selecting a single colony, inoculating the single colony on a seed culture medium for culture, and culturing at 30 ℃ for 24h to obtain fusarium oxysporum seed liquid for later use; the formula of the seed culture medium is as follows: 10g/L glucose, 6g/L yeast extract, 1g/L dipotassium phosphate, 1g/L potassium dihydrogen phosphate, 0.01g/L magnesium sulfate and 0.01g/L ferrous sulfate;
inoculating cladosporium dendritic on a PDA culture medium for culturing to obtain a single colony; selecting a single colony, inoculating the single colony to a PDA liquid culture medium for culture, and culturing at 30 ℃ for 24 hours to obtain a cladosporium dendritic seed solution for later use;
sequentially inoculating fusarium oxysporum seed liquid and cladosporium dendritic seed liquid into an MS culture medium, wherein the inoculation amounts are 5% and 4%, respectively, adding 70mg/L of 6-BA, 70mg/L of NAA and 30mg/L of DA-6, and stirring at 100rpm for 6 hours at the temperature of 28 ℃ to obtain the fusarium oxysporum seed liquid.
The fusarium oxysporum is ATCC 76222; the cladosporium cladosporioides is ATCC 34010.
Example 3
A stimulant for plant graft propagation, comprising a plant hormone and a fungus;
the preparation method specifically comprises the following steps:
inoculating fusarium oxysporum on a PDA culture medium for culturing by streaking to obtain a single colony; selecting a single colony, inoculating the single colony on a seed culture medium for culture, and culturing at 30 ℃ for 24h to obtain fusarium oxysporum seed liquid for later use; the formula of the seed culture medium is as follows: 10g/L glucose, 6g/L yeast extract, 1g/L dipotassium phosphate, 1g/L potassium dihydrogen phosphate, 0.01g/L magnesium sulfate and 0.01g/L ferrous sulfate;
inoculating cladosporium dendritic on a PDA culture medium for culturing to obtain a single colony; selecting a single colony, inoculating the single colony to a PDA liquid culture medium for culture, and culturing at 30 ℃ for 24 hours to obtain a cladosporium dendritic seed solution for later use;
sequentially inoculating fusarium oxysporum seed liquid and cladosporium dendritic seed liquid into an MS culture medium, wherein the inoculation amounts are 5% and 7%, respectively, adding 50mg/L of 6-BA, 40mg/L of NAA and 50mg/L of DA-6, and stirring at the temperature of 29 ℃ and 100rpm for 4 hours to obtain the fusarium oxysporum seed liquid.
The fusarium oxysporum is ATCC 76222; the cladosporium cladosporioides is ATCC 34010.
Example 4
Application example of the stimulant of the invention in grafting: take grafting of camellia as an example.
The method comprises the following steps of taking a common camellia oleifera large tree as a stock, grafting Yunnan camellia rich in peach red, averagely distributing 5 grafting stocks per plant, grafting 4 scions to the stock per branch, selecting the five-month bottom-six month beginning for grafting, and grafting by adopting a scion grafting method; sawing off the stock about 100cm away from the ground, and disinfecting the stock 2-3 days before grafting; during grafting, the lower part of the scion is soaked in an irritant for 30min, then taken out, grafted and then bundled for shading; counting the scion survival rate (the survival rate percent is calculated by the method of the number of the survival scions/the total number of the grafting scions) in 30 days, wherein the survival judgment standard is as follows: the grafted part heals and turns from red to green. And calculating the growth amount of the scion and the lateral bud extraction amount at the bottom of 10 months.
The components of the stimulant are subjected to a multifactorial test to verify the relationship between the plant growth hormone and the microorganism, which is shown in table 1:
TABLE 1
| Factors of the fact | Group 1 (example 1) | Group 2 | Group 3 | Group 4 | Group 5 |
| Fusarium pachysporium seed liquid | ＋ | － | ＋ | ＋ | － |
| Cladosporium cladonioides seed liquid | ＋ | ＋ | － | ＋ | － |
| 6-BA+ NAA+DA-6 | ＋ | ＋ | ＋ | － | ＋ |
The five groups of stimulants in the table 1 are respectively adopted for grafting, and the five groups of stimulants are divided into five test areas, and the operation modes are the same. 20 plants were randomly selected from each test area and examined, and the average index was calculated. The statistical results of the scion survival rate, the scion growth amount and the lateral bud shoot number of each group are shown in table 2:
TABLE 2
| Group of | Scion survivalPercentage ratio% | Growth of ear in cm | Number of lateral bud tips |
| Group 1 | 96.5 | 5.9 | 2.8 |
| Group 2 | 81.0 | 5.4 | 2.5 |
| Group 3 | 83.5 | 5.5 | 2.6 |
| Group 4 | 62.0 | 4.8 | 2.0 |
| Group 5 | 71.5 | 5.1 | 2.2 |
And (4) conclusion: as can be seen from table 2, compared with group 5 using only phytohormones, the scion survival rate of group 1 was increased by 25%, the scion survival rate of group 2 was increased by 9.5%, and the scion survival rate of group 3 was increased by 12%; group 4 only used microorganisms as the stimulator, which had a poor effect, which was lower than group 5 using plant growth hormone, and thus, the effect of stimulating by microorganisms alone or plant hormone was poor, and the addition of specific microorganisms to the stimulator increased the scion survival rate, which was the highest in group 1 and was the result of co-stimulation by two microorganisms and plant growth hormone; by observing two important indexes of the growth amount of the ear strips and the number of the lateral bud tips, the growth amount of the ear strips and the number of the lateral bud tips of the group 1 are higher than those of other groups, compared with a control group 5 only adopting plant growth hormone, the growth amount of the ear strips is improved by 0.8cm, and the number of the lateral bud tips is improved by 0.6. Probably, microorganisms and plant tissues form symbiosis, interact and influence with each other, good ecological balance is established, substances for promoting plant growth, such as gibberellin, auxin, cytokinin and the like, can be generated, the plant can absorb P, N and other nutrient elements, and meanwhile, the nutrient elements and space can be competed with pathogenic bacteria, so that the plant growth can be promoted.
Similar phenomena are found in grafting of other camellia plants, for example, the scion adopts a child surface which is difficult to graft and survive, the scion survival rate can be improved by more than 20 percent compared with the conventional operation, and other indexes such as the growth amount of scion strips, the lateral bud extraction amount and the like are improved; the grafting effect of the stimulant in other plant varieties is further researched subsequently.
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (4)
1. The stimulant for plant grafting propagation is prepared by the following process:
step 1) inoculating fusarium oxysporum on a PDA culture medium for culturing by streaking to obtain a single colony; selecting a single colony, inoculating the single colony on a seed culture medium for culture, and culturing at 30 ℃ for 24h to obtain fusarium oxysporum seed liquid for later use;
step 2) inoculating cladosporium dendritic on a PDA culture medium for culturing to obtain a single colony; selecting a single colony, inoculating the single colony to a PDA liquid culture medium for culture, and culturing at 30 ℃ for 24 hours to obtain a cladosporium dendritic seed solution for later use;
and 3) sequentially inoculating the fusarium oxysporum seed liquid and the cladosporium cladosporioides seed liquid into an MS culture medium, wherein the inoculation amounts are 3-5% and 4-7%, respectively, adding 6-BA, NAA and DA-6, and stirring at 100rpm for 3-6 hours at the temperature of 28-30 ℃ to obtain the fusarium oxysporum seed liquid.
2. The stimulant of claim 1, wherein the formulation of the seed culture medium is: 10g/L glucose, 6g/L yeast extract, 1g/L dipotassium phosphate, 1g/L potassium dihydrogen phosphate, 0.01g/L magnesium sulfate and 0.01g/L ferrous sulfate.
3. The stimulant of claim 1, wherein the Fusarium oxysporum is ATCC 76222; the cladosporium cladosporioides is ATCC 34010.
4. The stimulant according to claim 1, wherein the addition amount of 6-BA is 50-100mg/L, the addition amount of NAA is 40-70mg/L, and the addition amount of DA-6 is 30-50 mg/L.
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| CN103828722B (en) * | 2014-03-24 | 2015-06-24 | 鲁东大学 | Method for blueberries seedling and cultivating blueberries in large area by applying DSE (Dark Septate Endophyte) fungus |
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| CN103828722B (en) * | 2014-03-24 | 2015-06-24 | 鲁东大学 | Method for blueberries seedling and cultivating blueberries in large area by applying DSE (Dark Septate Endophyte) fungus |
| CN104250620A (en) * | 2014-09-24 | 2014-12-31 | 山东省农业科学院植物保护研究所 | Chlamydospore fusarium strain and application thereof |
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Denomination of invention: A stimulating agent for plant grafting and reproduction and its preparation method Effective date of registration: 20231220 Granted publication date: 20200619 Pledgee: Datian County Sub branch of Postal Savings Bank of China Co.,Ltd. Pledgor: FUJIAN SHANGUAGUA AGRICULTURAL DEVELOPMENT CO.,LTD. Registration number: Y2023350000253 |