CN108848990B - Method for cultivating hermaphrodite taxus chinensis by micro-grafting - Google Patents
Method for cultivating hermaphrodite taxus chinensis by micro-grafting Download PDFInfo
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- CN108848990B CN108848990B CN201810785651.4A CN201810785651A CN108848990B CN 108848990 B CN108848990 B CN 108848990B CN 201810785651 A CN201810785651 A CN 201810785651A CN 108848990 B CN108848990 B CN 108848990B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G2/00—Vegetative propagation
- A01G2/30—Grafting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
Abstract
The invention discloses a method for cultivating hermaphrodite taxus chinensis by microbody grafting, which comprises the following steps of primary culture: respectively inoculating the female plant seedlings and the male plant branches into culture bottles filled with a No. 1 culture medium and a No. 2 culture medium for culturing for 1-2 weeks; micro grafting: cutting off at least one terminal bud of a branch of a female stock; taking out the tassel of the male plant, cutting the base part flat, grafting the tassel of the male plant to the branch of the female plant seedling with the top bud removed; and (3) management after grafting: sealing the culture bottle, continuously culturing the female stock grafted with the male scion in the culture bottle filled with the No. 1 culture medium for 3-4 weeks, and promoting the healing of the grafting opening to obtain a grafted seedling; transplanting the grafted seedlings into a humus soil matrix, putting the humus soil matrix into a greenhouse for acclimation and culture for 3-4 weeks, and culturing to obtain hermaphrodite taxus chinensis seedlings. The hermaphrodite yew cultivated by the method for cultivating the hermaphrodite yew through the microbody grafting has both the female flowering branch and the male flowering branch, realizes the hermaphrodite yew, and improves the ornamental effect.
Description
Technical Field
The invention relates to the field of plant cultivation, in particular to a method for cultivating hermaphrodite taxus chinensis by micro-body grafting.
Background
Taxus cuspidata (Taxus cuspirata) belongs to Taxaceae (Taxaceae), and Taxus genus (Taxus) evergreen conifer trees. The taxus cuspidata is a natural rare anti-cancer plant which is recognized to be endangered and extincted in the world, has a history of 250 ten thousand years on the earth and is a plant activating stone. The United nations textbook organization listed the plants as rare or endangered plants in the world in 1996; the wild plants are listed as first-grade rare or endangered wild plants in China in 1999.
The greening and appreciation are one of the purposes of the taxus cuspidata, the taxus cuspidata is beautiful in tree shape, the branches and leaves are dense and jade in four seasons, fruits are watched in summer and autumn, berries are bright red, ruby is embedded in huge jade, and the trees are rare and green ornamental trees in the northeast. And the northeast yew is male and female, pollen has no air pocket, the flying distance is short, and the optimal pollination period is short. Only by culturing the male and female taxus cuspidata plants at the same time, fruits can be produced, otherwise, the fruits cannot be fruited, and the purpose of viewing the fruits is difficult to realize.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a method for cultivating hermaphrodite taxus chinensis by micro-grafting, which aims to solve the problem that a single plant cannot bear fruits.
In order to achieve the purpose of the invention, the invention adopts the following technical scheme:
the invention aims to provide a method for cultivating hermaphrodite taxus chinensis by micro-body grafting, which comprises the following steps:
s1, primary culture: under the aseptic condition, inoculating female plant seedlings into a culture bottle filled with a No. 1 culture medium for culturing for 1-2 weeks, inoculating male plant branches into a culture bottle filled with a No. 2 culture medium for culturing for 1-2 weeks, and respectively carrying out primary culture on female plant stocks and male plant scions;
s2, micro grafting:
under the aseptic condition, cutting off at least one terminal bud of a branch of a female stock to prepare for grafting;
taking out the male plant scion under the aseptic condition, cutting the base part flat, grafting the male plant scion to the branch of the female plant seedling with the terminal bud removed, and finishing grafting;
s3, management after grafting:
sealing the culture bottle under aseptic conditions, and continuously culturing the female stock grafted with the male scion in the culture bottle filled with the No. 1 culture medium for 3-4 weeks to heal the grafting opening, so as to obtain a grafted seedling;
transplanting the grafted seedlings into a humus soil matrix, putting the humus soil matrix into a greenhouse for acclimation and culture for 3-4 weeks, and culturing to obtain hermaphrodite taxus chinensis seedlings.
Preferably, the primary culture of S1 comprises the following steps:
culturing female stock: selecting a female taxus chinensis plant, shearing 1-2 year-old branches, keeping 3-5 small branches, quickly dipping ABT rooting powder, cutting, controlling the temperature to be 20 +/-2 ℃ and the relative humidity to be 90-100% by using humus as a cutting medium, and inducing adventitious roots, wherein the length of the branches is 3-6 cm; culturing for 6-7 weeks, taking roots successively, sterilizing the inner surface of an ultra-clean workbench in a sterile room when the root length is 0.3-0.5 cm, inoculating the sterilized inner surface into a culture bottle filled with the No. 1 culture medium under the sterile condition, placing the culture bottle filled with the No. 1 culture medium in the culture room for culturing for 1-2 weeks, and removing stain strains to obtain female stock for grafting;
culturing the male plant scions: selecting male plant branches with the diameter similar to that of the female branches, sterilizing the male plant branches on the inner surface of an ultra-clean workbench in a sterile room, inoculating the male plant branches into a culture bottle filled with a No. 2 culture medium under the sterile condition, placing the culture bottle filled with the No. 2 culture medium in a culture room for culturing for 1-2 weeks, and removing stain strains to obtain the male plant scions for grafting.
Preferably, the medium No. 1 comprises modified MS medium, 6-benzylamino adenine, 2,4-D and naphthylacetic acid; the No. 2 culture medium comprises an improved MS culture medium, 6-benzylamino adenine and naphthylacetic acid.
Preferably, the grafting of S2 specifically comprises the following steps:
selecting female stock with good growth, shearing 1-2 branch terminal buds in an ultraclean workbench under an aseptic condition, and preparing for grafting;
taking out the male plant scion under aseptic condition, cutting the base part flat, grafting the male plant scion to the branch of the female plant seedling with the terminal bud removed, and completing grafting.
Preferably, 1-2 branch terminal buds are cut off by using a bent scissors, so that the cut section of the branch is a horizontal section.
Preferably, the bent-type scissors have a V-shape with two extending ends, and at least one of the extending ends has a vertical bending cut portion at an end thereof.
Preferably, the specific steps of grafting the tassel of the male plant to the branch of the female plant seedling with the terminal bud removed are as follows:
under aseptic condition, the cut plane of the base part of the scion of the male plant is contacted with the horizontal section of the female plant seedling with the top bud of the branch cut off, so as to complete grafting.
Preferably, the specific steps of S3 post-grafting management are:
sealing the culture bottle under aseptic conditions, and continuously culturing the female stock grafted with the male scion in the culture bottle filled with the No. 1 culture medium for 3-4 weeks to heal the grafting opening, so as to obtain a grafted seedling;
opening a culture bottle mouth, performing adaptive exercise on the grafted seedling, taking out the grafted seedling after 5-7 days, and cleaning a culture medium adhered to the root of the grafted seedling;
and (3) carrying out traditional Chinese medicine bath on the grafted seedlings in a bactericide aqueous solution, then transplanting the grafted seedlings into a humus soil matrix, putting the humus soil matrix into a greenhouse for hardening seedlings, carrying out acclimation culture for 3-4 weeks, and then culturing to obtain the hermaphrodite taxus chinensis seedlings.
Preferably, the temperature is controlled at 22 +/-2 ℃ during the seedling training period; and (3) gradually reducing the relative humidity of air after the relative humidity of the air is reduced by 80-90% at the initial stage of seedling training, and spraying a bactericide once every 5 days to prevent the grafted seedlings from infecting mixed bacteria.
Preferably, the bactericide is carbendazim.
Compared with the prior art, the hermaphroditic yew cultivated by the method for cultivating the hermaphroditic yew through the microbody grafting has the advantages that the hermaphroditic yew has both female flowering branches and male flowering branches, the hermaphroditic yew is realized, and the ornamental effect is improved.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise.
FIG. 1 shows a female stock being bred;
FIG. 2 shows a scion of a male plant;
FIG. 3 is a diagram showing the seedlings of the same male and female yew cultivated by the method for cultivating the same male and female yew by microbody grafting of the present invention;
FIG. 4 is a photograph of a cut out of the terminal bud of a female stock using a pair of curved scissors.
Detailed Description
Technical solutions in the embodiments of the present invention will be described in detail below with reference to the drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only a part of the embodiments of the present invention, and not all the 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 scope of the present invention.
The invention provides a method for cultivating a hermaphroditic taxus chinensis by micro-grafting, which takes female plant seedlings as stocks and male plant branches as scions for micro-grafting, and specifically comprises the following steps:
s1, primary culture: under the aseptic condition, the female plant seedlings are inoculated into a culture bottle filled with a No. 1 culture medium for culture for 1-2 weeks, the male plant branches are inoculated into a culture bottle filled with a No. 2 culture medium for culture for 1-2 weeks under the aseptic condition, and primary culture is respectively carried out on female plant stocks and male plant scions.
The medium No. 1 in the culture flask with medium No. 1 therein comprises modified MS medium, 6-benzylamino adenine, 2,4-D and naphthylacetic acid. Preferably, the concentration of 6-benzylaminopurine is 0.3-0.6 mg/L, the concentration of 2,4-D is 0.05-0.15 mg/L, and the concentration of naphthylacetic acid is 0.1-0.3 mg/L. Preferably, medium No. 1 includes modified MS medium, 0.5 mg/L6-benzylamino adenine, 0.1 mg/L2, 4-D and 0.3mg/L naphthylacetic acid.
The No. 2 medium in the culture flask with the No. 2 medium therein comprises modified MS medium, 6-benzylamino adenine and naphthylacetic acid. Preferably, the content of 6-benzylaminopurine is 0.3-0.6 mg/L, and the content of naphthylacetic acid is 0.1-0.3 mg/L; preferably, Medium No. 2 includes modified MS Medium, 0.5 mg/L6-benzylamino adenine and 0.2mg/L naphthylacetic acid.
Specifically, the specific steps of culturing the rootstock and the scion in the S1 are as follows:
culturing female stock: referring to the figure 1, selecting a female taxus chinensis plant, shearing 1-2 year-old branches, shearing the branches with the length of 3-6 cm, reserving 3-5 small branches, dipping ABT rooting powder quickly, cutting, controlling the temperature to be 20 +/-2 ℃, controlling the relative humidity to be 90-100% and inducing adventitious roots, wherein the cutting medium is humus soil; culturing for 6-7 weeks, taking roots successively, sterilizing the inner surface of an ultra-clean workbench in a sterile room when the root length is 0.3-0.5 cm, inoculating the sterilized inner surface into a culture bottle filled with the No. 1 culture medium under the sterile condition, placing the culture bottle filled with the No. 1 culture medium in the culture room for culturing for 1-2 weeks, and removing stain strains to obtain female stock for grafting;
culturing the male plant scions: referring to fig. 2, selecting male plant branches with the diameter similar to that of the female branches, sterilizing the male plant branches on the inner surface of an ultra-clean workbench in a sterile room, inoculating the male plant branches into a culture bottle filled with a No. 2 culture medium under the sterile condition, placing the culture bottle filled with the No. 2 culture medium in a culture room for culturing for 1-2 weeks, and removing the stain to obtain the male plant scion for grafting.
S2, micro grafting;
under the aseptic condition, cutting off at least one terminal bud of a branch of a female stock to prepare for grafting;
taking out the male plant scion under aseptic condition, cutting the base part, grafting the male plant scion to the branch of the female plant seedling with the terminal bud removed, and completing grafting.
Specifically, the grafting in S2 specifically comprises the following steps:
selecting female stock with good growth, shearing 1-2 branch terminal buds in an ultraclean workbench under an aseptic condition, and preparing for grafting;
taking out the male plant scion under aseptic condition, cutting the base part flat, grafting the male plant scion to the branch of the female plant seedling with the terminal bud removed, and completing grafting. Specifically, under aseptic conditions, the cut plane of the scion base of the male plant is contacted with the horizontal section of the female plant seedling from which the terminal bud of the branch is cut, so as to complete grafting.
Referring to fig. 4, 1-2 terminal buds of the branches are cut off, and the branches are cut by a bent scissors, so that the cut sections of the branches are horizontal sections.
Furthermore, the bent scissors are V-shaped with two extending ends, and the end part of at least one extending end is provided with a vertical bending cutting part.
S3, managing after grafting;
specifically, referring to fig. 3, the specific steps of post-grafting management in S3 are as follows:
s31, sealing the culture bottle, and culturing the female stock grafted with the male scion in the culture bottle filled with the No. 1 culture medium for 3-4 weeks under the aseptic condition to heal the grafting opening to obtain a grafted seedling;
s32, opening a culture bottle mouth, performing adaptive exercise on the grafted seedling, taking out the grafted seedling after 5-7 days, and cleaning a culture medium adhered to the root of the grafted seedling;
s33, the grafted seedlings are subjected to Chinese medicine bath in a bactericide aqueous solution, then the grafted seedlings are transplanted into a humus soil matrix, the humus soil matrix is placed in a greenhouse for hardening seedlings, and after acclimation culture is carried out for 3-4 weeks, the seedlings of the hermaphrodite taxus chinensis are obtained through cultivation.
Wherein the temperature is controlled to be 22 +/-2 ℃ during the seedling training period; and (3) gradually reducing the relative humidity of air after the relative humidity of the air is reduced by 80-90% at the initial stage of seedling training, and spraying a bactericide once every 5 days to prevent the grafted seedlings from infecting mixed bacteria.
Preferably, the bactericide is carbendazim.
Example 1
In the embodiment, a female taxus cuspidata plant is selected, 1-year-old branches are cut, the length of each branch is 4-5 cm, 3-5 small branches are reserved, ABT rooting powder is dipped quickly, cuttage is carried out, humus soil is used as a cuttage substrate, the temperature is controlled to be 20 +/-2 ℃, the relative humidity is 90-100%, and adventitious roots are induced.
Culturing for 6-7 weeks, sequentially rooting, sterilizing the inner surface of an ultra-clean workbench in a sterile room when the root length is 0.3-0.5 cm, transplanting the seedlings into a prepared MS culture medium under the sterile condition, and culturing in a culture room.
Meanwhile, selecting male plant branches with the same diameter, sterilizing the branches on the inner surface of a clean room superclean workbench, inoculating the branches into a MS culture medium prepared in advance under a sterile condition, and culturing the scions in a culture room. Culturing the female seedlings and the male scions in an MS culture medium for 1-2 weeks, and removing the infected strains.
Selecting well-grown female seedlings, shearing 1-2 branches with a special bent type scissors under the aseptic condition in an ultra-clean workbench, taking out male scion strips, shearing the base parts, grafting the male scion strips on the broken branches of the female seedlings, sealing a bottle cover, and putting the bottle cover back on a culture frame for continuous culture.
Culturing for 3-4 weeks until the grafting opening is healed, removing the bottle cap in a culture room, performing adaptive exercise on the grafted seedling, taking out the grafted seedling after 5-7 days, cleaning the culture medium adhered to the root, performing traditional Chinese medicine bath in 800 times of carbendazim aqueous solution, transplanting the grafted seedling into a humus matrix, and putting the humus matrix in a greenhouse for hardening the seedling.
Controlling the temperature to be 22 +/-2 ℃ during the seedling training period; the relative humidity of the seedlings in the initial stage of seedling exercising is 80-90%, and then the relative humidity of air is gradually reduced; spraying the bactericide once every 5 days to prevent the seedlings from infecting mixed bacteria. Domesticating and culturing in a greenhouse for 3-4 weeks to realize natural growth, wherein the management measures are the same as those of taxus cuspidata cutting seedlings. Thus, the cultivation of the homosexual seedlings of the taxus cuspidata is successful.
The hermaphrodite yew cultivated by the method for cultivating the hermaphrodite yew through the microbody grafting has both the female flowering branch and the male flowering branch, realizes the hermaphrodite yew, and improves the ornamental effect.
While the invention has been shown and described with reference to certain embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims and their equivalents.
Claims (6)
1. A method for cultivating hermaphrodite taxus chinensis by micro-grafting is characterized by comprising the following steps:
s1, primary culture; the S1 primary culture comprises the following specific steps:
culturing female stock: selecting a female taxus chinensis plant, shearing 1-2 year-old branches, keeping 3-5 small branches, quickly dipping ABT rooting powder, cutting, controlling the temperature to be 20 +/-2 ℃ and the relative humidity to be 90-100% by using humus as a cutting medium, and inducing adventitious roots, wherein the length of the branches is 3-6 cm; culturing for 6-7 weeks, taking roots successively, sterilizing the inner surface of an ultra-clean workbench in a sterile room when the root length is 0.3-0.5 cm, inoculating the sterilized inner surface into a culture bottle filled with the No. 1 culture medium under the sterile condition, placing the culture bottle filled with the No. 1 culture medium in the culture room for culturing for 1-2 weeks, and removing stain strains to obtain female stock for grafting; wherein the No. 1 culture medium comprises a modified MS culture medium, 6-benzylamino adenine, 2,4-D and naphthylacetic acid;
culturing the male plant scions: selecting male plant branches with the diameter similar to that of the female branches, sterilizing the male plant branches on the inner surface of an ultra-clean workbench in a sterile room, inoculating the male plant branches into a culture bottle filled with a No. 2 culture medium under the sterile condition, placing the culture bottle filled with the No. 2 culture medium in a culture room for culturing for 1-2 weeks, and removing stain strains to obtain male plant scions for grafting; wherein the No. 2 culture medium comprises an improved MS culture medium, 6-benzylamino adenine and naphthylacetic acid;
s2, micro grafting; the grafting of S2 comprises the following specific steps:
selecting female stock with good growth, shearing 1-2 branch terminal buds in an ultraclean workbench under an aseptic condition, and preparing for grafting;
taking out the male plant scion under the aseptic condition, cutting the base part flat, grafting the male plant scion to the branch of the female plant seedling with the terminal bud removed, and finishing grafting;
s3, managing after grafting; the specific steps of the management after S3 grafting are as follows:
sealing the culture bottle under aseptic conditions, and continuously culturing the female stock grafted with the male scion in the culture bottle filled with the No. 1 culture medium for 3-4 weeks to heal the grafting opening, so as to obtain a grafted seedling;
opening a culture bottle mouth, performing adaptive exercise on the grafted seedling, taking out the grafted seedling after 5-7 days, and cleaning a culture medium adhered to the root of the grafted seedling;
and (3) carrying out traditional Chinese medicine bath on the grafted seedlings in a bactericide aqueous solution, then transplanting the grafted seedlings into a humus soil matrix, putting the humus soil matrix into a greenhouse for hardening seedlings, carrying out acclimation culture for 3-4 weeks, and then culturing to obtain the hermaphrodite taxus chinensis seedlings.
2. The method for cultivating the hermaphroditic yew by the micro-body grafting according to claim 1, wherein 1-2 branch terminal buds are cut by a bent scissors, so that the section of the cut branch is a horizontal section.
3. The method for cultivating the hermaphroditic yew by micro-grafting according to claim 2, wherein the curved scissors have a V shape with two extending ends, and the end of at least one extending end has a vertical bending cut part.
4. The method for cultivating the hermaphroditic taxus chinensis by micro-body grafting according to claim 2, wherein the specific steps of grafting the tassel of the male plant to the branch of the female plant seedling with the terminal bud cut off are as follows:
under aseptic condition, the cut plane of the base part of the scion of the male plant is contacted with the horizontal section of the female plant seedling with the top bud of the branch cut off, so as to complete grafting.
5. The method for cultivating the hermaphrodite taxus chinensis by micro-body grafting according to claim 1, wherein the temperature is controlled to be 22 +/-2 ℃ during seedling hardening; and (3) gradually reducing the relative humidity of air after the relative humidity of the air is 80-90% at the initial stage of seedling hardening, and spraying a bactericide once every 5 days to prevent the grafted seedlings from infecting mixed bacteria.
6. The method for cultivating the hermaphrodite taxus chinensis by micro-body grafting according to claim 1, wherein the bactericide is carbendazim.
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| CN105746356A (en) * | 2016-03-21 | 2016-07-13 | 广西壮族自治区农业科学院花卉研究所 | Distant test tube grafting method for Chinese roses and wild roses |
| CN106417025A (en) * | 2016-10-14 | 2017-02-22 | 山西农业大学 | Test tube micro-grafting method for seedless grape embryo-rescued malformed plantlets |
| CN106718883A (en) * | 2016-11-28 | 2017-05-31 | 华中农业大学 | A kind of Yunnan Chinese catalpa and the miniature engrafting method of test tube seedling of Chinese catalpa |
| CN107211796A (en) * | 2017-06-18 | 2017-09-29 | 余颖恒 | A kind of root system propagation method of Chinese yew |
| CN107278633A (en) * | 2017-08-05 | 2017-10-24 | 崇义县绿地种苗场 | A kind of high position grafting method of Chinese yew |
| CN107509536A (en) * | 2017-10-18 | 2017-12-26 | 广州市农业科学研究院 | A kind of outer micro-grafting method of test tube of balsam pear tissue-cultured seedling |
| CN107821080A (en) * | 2017-11-27 | 2018-03-23 | 四川农业大学 | A kind of method for promoting Taxus x media potted landscape itself to yield positive results |
| CN108243756A (en) * | 2018-03-20 | 2018-07-06 | 山东农业大学 | A kind of Tissue-cultured apple seedling bottle grafts the micro-grafting method taken root outside |
| CN108260427A (en) * | 2018-04-24 | 2018-07-10 | 大理弥纳食品有限公司 | Quick engrafting method in a kind of rose room |
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