CN103039362A - Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof - Google Patents

Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof Download PDF

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CN103039362A
CN103039362A CN2013100129925A CN201310012992A CN103039362A CN 103039362 A CN103039362 A CN 103039362A CN 2013100129925 A CN2013100129925 A CN 2013100129925A CN 201310012992 A CN201310012992 A CN 201310012992A CN 103039362 A CN103039362 A CN 103039362A
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culture
seedling
subculture
inoculation
oil tea
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CN103039362B (en
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刘越秀
潘新建
周其新
张运斌
赵忠菊
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HUANGSHAN INSTITUTE OF FORESTRY
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HUANGSHAN INSTITUTE OF FORESTRY
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Abstract

The invention discloses a subculture medium for tissue culture seedling propagation of camellia oleifera abel, which comprises 0.6-3.0mg/L MS+6-BA and 0.06-1.5mg/L IAA. A subculture medium formed by adding 6-benzyl aminopurine and indoleacetic acid with specific concentration into an MS minimal medium performs subculture of the aseptic seedlings of camellia oleifera abel, and the multiplication rate of the aseptic seedlings of camellia oleifera abel can reach 3.88-5.57 after 40 days; and meanwhile, strong seedling treatment of the camellia oleifera abel seedlings is realized, subsequent strong seedling culture is not required for culturing the aseptic seedlings of camellia oleifera abel, the propagation period of tissue culture seedlings is shortened, the technology is simplified, and the raw materials for configuring a strong seedling medium are saved.

Description

The breeding of oil tea group training seedling subculture medium and propagation method thereof
Technical field
The present invention relates to the vegetative propagation field, be specifically related to a kind of breeding of oil tea group training seedling subculture medium and propagation method thereof.
Background technology
Oil tea is the distinctive high-quality woody oil tree species of China, at the cultivation history in existing more than 2300 year of China oil tea.The tea oil look that it the squeezes perfume (or spice) of distinguishing the flavor of clearly, nutritious, the cardiovascular and cerebrovascular diseases such as hypertension, high fat of blood, cardiopathy are had good medical care effect, united state classifies the senior edible vegetable oil of health type that emphasis is promoted as.Simultaneously, the by-product of tea oil is sampled tea withered and the tea shell can extract Tea Saponin by comprehensive utilization, grind polishing powder and fermentation high protein feed etc., and oil-tea camellia husks can be produced activated carbon and extract tanning material etc., so the added value of oil tea is high.In addition, oil tea wide adaptability, barren-resistant is one of low mountain, south, the good nonwood forest trees in hills and hillock, and does not strive ground with grain and cotton that tool is planted one year, and the characteristics of income are called " iron crops ", " green oil depot " by the self-employed tree cultivator for many years.Therefore, the development camellia oleiferaindustry not only is conducive to alleviate the situation of the long-term dependence on import of China's edible oil, supply and demand anxiety, and the economic benefit of oil tea is high simultaneously, can increase peasant's income, the development that promotes rural economy and the national general level of the health of raising.
The vegetative propagation technique measures such as main employing cuttage and seedling culture, sprout stock grafting and the changing-over of low production forest height of breeding of oil tea seedling.Yet, owing to be subject to the limited restriction of oil tea improved seeds clonal reproduction material resources, add the factor affecting such as the cultivating seedlings time is long, the seedling propagation coefficient is low, being difficult at short notice, quickly breeding goes out a large amount of choiceness nursery stocks, to satisfy the demand of camellia oleiferaindustry development, therefore, active demand provides the propagation method of the oil tea group training seedling that a kind of reproduction coefficient is high, transplanting survival rate is high.
At present, study comparatively " research of oil tea cultured in vitro regeneration induction plant " (Bi Fang tomahawk etc. of famous being called of report in forward position about the breeding of oil tea group training seedling, economic forest research, the 2nd phase of the 22nd volume, the 5th~9 page, 2004) scientific and technical literature, it discloses with stem section, rataria, the cotyledon of Camellia Oleifera Clones Hunan woods No. 4 and has induced breeding in vitro, the stem section is cultivated take MS as minimal medium, additional 6-BA3.0mg/L+NAA1.0mg/L induces bud, and inductivity reaches 83.33%; Continue to carry out on the former medium subculture cultivation and obtain Multiple Buds, for rataria and cotyledon, additional hormone 2,4~D2.0mg/L+KT1.0mg/L induces, the callus of inducing generation is forwarded on the medium of additional 6-BA3.0mg/L+NAA0.05mg/L and 6-BA2.5mg/L+IAA1.5mg/L and induce indefinite bud, inductivity reaches more than 90%, and further disclose aseptic seedling root induction on the MS+NAA7.0mg/L medium the suitableeest, but the regrowth that adopts its disclosed medium to carry out the root induction acquisition only has a main root mostly, fibrous root seldom, the difficulty of taking root after the group training transplantation of seedlings, be difficult to survive, therefore, its in fact and fail to obtain can transplant survival group training seedling.Simultaneously, the concentration of 6-BA, IAA in its inducing culture that disposes is higher, induces the effect of sprouting also bad in actual mechanical process.
In addition, name is called and discloses following technical scheme in " a kind of efficient rooting method for oil tea clone tissue culture seedlings " Chinese patent literature (CN102007867A): efficient rooting method for oil tea clone tissue culture seedlings, comprise just culture, subculture cultivation, strong seedling culture and process of rooting culture, wherein after the strong seedling culture, need dipping pretreatment 5~10s in the solution of plant hormone IBA1000~4000ppm, then be seeded in and cultivate the oil tea group training seedling that obtains to have root system on the root media.Although adopt this scheme can obtain oil tea group training seedling, but experimental study shows, the IBA solution that each aseptic seedling all must more renew before taking root and inoculating soaks it, the solution that is used for the immersion aseptic seedling can not be reused, otherwise the infection of the as easy as rolling off a log occurrence of large-area of aseptic seedling causes the waste of aseptic seedling death and raw material.Simultaneously, the medium that is used for the subculture cultivation that it provides can only promote aseptic seedling to breed, and can't promote Aseptic Seedling Growth (strong sprout), therefore also needs to carry out strong seedling culture in its follow-up phase, so that the man-hour of whole group of training seedling breeding is long.In addition, bud induction rate and the rooting rate in the process of rooting culture in its first culture is still lower.
Summary of the invention
Primary and foremost purpose of the present invention provides the breeding of a kind of oil tea group training seedling and uses subculture medium, can realize simultaneously propagation and the strong sprout of oil tea aseptic seedling when subculture is cultivated, and reduces the breeding cycle of oil tea group training seedling.
Its scheme of taking is: subculture medium is used in the breeding of a kind of oil tea group training seedling, it is characterized in that: comprise MS+6-BA0.6~3.0mg/L+IAA0.06~1.5mg/L.
By in the such scheme 6-benzyl aminopurine (6-BA) of interpolation certain concentration in the MS minimal medium and the subculture medium that heteroauxin (IAA) consists of being carried out the subculture cultivation to the oil tea aseptic seedling, the propagation multiple of oil tea aseptic seedling can reach 3.88~5.57 after 40 days, realize simultaneously processing the strong sprout of oil tea seedling, do not need the follow-up strong seedling culture that arranges again that the oil tea aseptic seedling is cultivated, shorten breeding cycle, the simplification technique of group training seedling and save the raw material that disposes the strong seedling culture base.
Another object of the present invention provides a kind of propagation method of oil tea group training seedling, a kind of propagation method of oil tea group training seedling, it is characterized in that: comprise that the stem section that intercepting sterilization on the oil tea plant is obtained is carried out first culture, subculture is cultivated and the aseptic seedling after the subculture cultivation is carried out culture of rootage, it is MS+6-BA0.6~3.0mg/L+IAA0.06~1.5mg/L that described subculture is cultivated used subculture medium.
By the effect of matching of above-mentioned group of training scheme and subculture medium, can obtain in the short period of time a large amount of oil tea group training seedlings.
Further technical scheme is that the used root media of described culture of rootage is 1/2~1/8MS+IBA2.4~4.8mg/L+NAA2.2~4.6mg/L among the application.The described just used first culture base of culture is MS+6-BA0.6~1.0mg/L+IAA0.07~1.3mg/L.Adopting above-mentioned root media to cultivate can be so that the rooting rate of oil tea aseptic seedling reaches 60~91.4%, average every strain aseptic seedling can be given birth to 1.5~3.18 root systems, root length can reach about 2.8cm, and the root system quality of the group of acquisition training seedling is good, thereby improves the transplanting survival rate of oil tea group training seedling.Simultaneously, be the amount reproduction of realization group training seedling, can carry out a subculture and cultivate, and then the Multiple Buds that will once cultivate intercepts into respectively the aseptic seedling that contains a tender shoots and carries out the secondary subculture and cultivate the growth coefficient of cultivating to improve subculture.
Wherein, MS refers to the minimal medium that is usually used in the breeding of plant tissue culture seedling according to relevant criterion or handbook or textbook configuration, 1/2~1/8MS refer to macroelement be by 1/2~1/8 times of macroelement concentration in the standard MS medium that disposes in handbook or standard or the textbook, trace element with by micro-identical medium in the standard MS medium that disposes in handbook or standard or the textbook.The concentration of IBA in the medium after IBA2.4~4.8mg/L refers to configure is 2.4~4.8mg/L, and the above-mentioned form of presentation that adopt in other places among the application all should so be understood.Certainly, adopting for those skilled in the art mineral salt commonly used, organic matter that cited raw material mineral salt, the organic matter of configuration MS minimal medium in handbook or standard or the textbook is equal to the concentration of replacing or changing among a small circle certain mineral salt and the MS minimal medium is cultivated the scheme that large change do not occur performance also should belong in the scope of MS minimal medium indication among the application.Among the application, preferably adopt potassium nitrate (KNO 3), ammonium nitrate (NH 4NO 3), magnesium sulfate (MgSO 4.7H 2O), potassium dihydrogen phosphate (KH 2PO 4), calcium chloride (GaCl 2), zinc sulphate (ZnSO 4﹒ 7H 2O), boric acid (H 3BO 3), sodium molybdate (NaMo0 4﹒ 5H 2O), copper sulphate (CuSO 4﹒ 5H 2O), cobalt chloride (CoCl 2﹒ 6H 20), the organic compound configuration MS minimal mediums such as the mineral salt such as potassium iodide (KI) and sucrose, vitamin, amino acid.
This propagation method also comprise will obtain after the culture of rootage group training transplantation of seedlings carry out hardening to the hardening matrix and process, hardening matrix is mixed by 28~32 weight portion sawdust/rice husks, 23~27 weight portion yellow soils, 4~6 weight portion ash, 28~32 weight portion northeast peat, 4~6 weight portion perlites and 4~6 weight portion vermiculites and is consisted of.Because the survival rate that the selected matrix of follow-up transplanting is trained transplantation of seedlings to group affects huge, the applicant finds by long term test, to above-mentioned hardening matrix, carry out hardening by organizing the training transplantation of seedlings, oil tea group training seedling can healthy and strong be grown, survival rate is high, improves the survival rate that follow-up oil tea seedling is gone up a hill and transplanted.
Concrete intercepting inoculation stem section and just culture, subculture cultivation, culture of rootage operation are respectively:
Described inoculation stem section is the inoculation branch of the robust growth that intercepts from tea oil tree, every blade on the inoculation branch is clipped 3/4~5/6 carry out clipping remaining blade and an inoculation branch base portion behind the cleaning and sterilizing again, then intercepts by an axillalry bud one stem section to obtain.
First culture: the inoculation stem section after will sterilizing is seeded in the inoculation bottle that is provided with first culture base, and inoculation bottle tightened with sealed membrane sealing, again the inoculation bottle lucifuge was processed 4~6 days, then carried out illumination cultivation 30~40 days, first culture base is MS+6-BA0.6~1.0mg/L+IAA0.07~1.3mg/L;
Subculture is cultivated: the aseptic seedling that first culture is obtained is transferred to carries out strong sprout and propagation is cultivated simultaneously in the inoculation bottle that is provided with subculture medium, cultivated 35~45 days, and subculture medium is MS+6-BA0.6~3.0mg/L+IAA0.06~1.5mg/L;
Culture of rootage: subculture is cultivated the Multiple Buds obtain be cut into 1.2~2.0cm and contain respectively the aseptic seedling of tender shoots and be seeded to cultivate in the inoculation bottle that is provided with root media and take root, cultivated 25~35 days, root media is 1/2~1/8MS+IBA2.4~4.8mg/L+NAA2.2~4.6mg/L.
Implement technical scheme of the present invention by the parameter of combinations thereof, just the bud induction rate of culture can reach 93.3%, the growth coefficient of aseptic seedling can reach more than 5.0 during subculture was cultivated, the rooting rate of culture of rootage can reach 91.4%, every strain group training seedling on average has 3.18 root systems, increases significantly qualitatively at reproduction coefficient and the group training seedling that obtains with respect to the other technologies parameter scheme that realizes the object of the invention.
Description of drawings
Fig. 1 a, 1b are first culture;
Fig. 2 a, 2b are that subculture is cultivated;
Fig. 3 a, 3b are culture of rootage;
Fig. 4 a, 4b, 4c are that hardening is processed.
Embodiment
To pass through embodiment, next the present invention is further illustrated.Wherein operating 1 is that the various medium of configuration are bred with inoculation stem section with obtaining, operation 2 is just in the culture base inoculation stem section to be carried out just culture in difference, operation 3 is to carry out the subculture cultivation to obtaining aseptic seedling behind the first culture in different subculture mediums, operation 4 is in different root medias subculture to be cultivated the rear aseptic seedling that obtains to carry out culture of rootage, and operation 5 is in different hardening matrix culture of rootage acquisition group training seedling to be carried out hardening to process.
Operation 1: the configuration medium is used the stem section with obtaining to breed
(a) configuration medium
It is stand-by to take by weighing respectively following mineral salt, organic matter, plant hormone and Organic additives;
Mineral salt: macroelement comprises potassium nitrate (KNO 3), ammonium nitrate (NH 4NO 3), magnesium sulfate (MgSO 4.7H 2O), potassium dihydrogen phosphate (KH 2PO 4), calcium chloride (GaCl 2) etc.; Trace element comprises zinc sulphate (ZnSO 4﹒ 7H 2O), boric acid (H 3BO 3), sodium molybdate (NaMo0 4﹒ 5H 2O), copper sulphate (CuSO 4﹒ 5H 2O), cobalt chloride (CoCl 2﹒ 6H 20), potassium iodide (KI) etc.;
Organic matter: comprise sucrose, vitamins and amino acid etc.;
Plant hormone: growth hormone heteroauxin (IAA), indolebutyric acid (IBA), methyl α-naphthyl acetate (NAA) are arranged, basic element of cell division 6-benzyl aminopurine (6-BA) and gibberellin.
Organic additives: Organic additives comprises artificial synthetic or natural organic matter, such as fruit juice, carragheen.
By the arrangements MS minimal medium of putting down in writing in relevant criterion or operation manual or the textbook, then get respectively the plant hormone of variable concentrations and kind and first culture base, subculture medium and the root media that the MS minimal medium is configured to various models, and stand-by after the sterilization processing.
(b) obtain breeding with inoculating the stem section
Inoculation material gathers: be taken from Mt. Huang in Anhui city institute of woods section oil tea breeding base for the examination material, kind has Mount Huang No. 1, No. 2, No. 4 and six kinds such as long woods No. 4, No. 27, No. 56.Acquisition time is in April~August, be selected in fine without the dew time period, with tree body middle and upper part give birth to then without infect, without the tender tip of damage by disease and insect, robust growth, as the first culture material of oil tea explant.The oil tea explant of adopting back is divided into four parts, and three parts install sealing with the plastics freshness protection package, are housed in 3~5 ℃ the refrigerator standby not same date inoculation survival rate comparative trial.
The processing of inoculation branch: every blade will inoculating on the branch (explant) cuts off 3/4~5/6, puts into beaker and washes secondary with the laundry vacuole, and then with clear water flush away liquid detergent, cushion 1h with running water after cleaning again.The oil tea inoculation branch drop water purification of buffering 1h is put on the biological cleaning workbench, and with removing alcohol behind 75% the alcohol-pickled 30s, the mercuric chloride solution of pouring fast again proportioning 1 ‰ into soaks sterilization 10~20min, determines sterilization time according to the tender degree of oil tea inoculation branch.During sterilization container is jiggled, make each position of inoculation branch all touch mercuric chloride solution, namely remove mercuric chloride solution after the sterilization, use again the sterile water continuous flushing more than 8 times, rock for every time about flushing 30s, then blot and cut off remaining blade with aseptic blotting paper and clip the base portion of inoculating branch, then by inoculating the branch bottom to ending, intercept take an axillalry bud as a stem section, stand-by as an inoculation stem section.
Operation 2: first culture
Transfer room and equipment disinfection sterilization: first the uviol lamp of transfer room is opened sterilization 30min, connect afterwards the clean work station power supply, start fan blower, light switch, infrared oese sterilizer, the alcohol spraying clean work station inwall with 75%; 75% alcohol is cleaned scissors, tweezers, and puts into 50 ℃~825 ℃ infrared oese sterilizer high-temperature sterilizations, takes out cooling after reaching sterilization effect; Be placed on the clean work station sterile water, aseptic paper, the aseptic blotting paper that disinfect for subsequent use.
Inoculation: the inoculation stem section that will intercept is inoculated in the inoculation bottle of the first culture base that contains various models as shown in table 1, and numbers just 1 group~first 6 groups, and 10 bottles every group, 3 inoculations of every bottle graft kind stem section, sealed membrane is tightened bottleneck, places culturing room to cultivate.Just the pickup kind is cultivated needs start illumination in dark place, greenhouse reason after 5 days, 25~28 ℃ of culturing room's temperature, humidity 60%, every day light application time 10h, intensity of illumination 1000~1500LX.
Observe the first culture situation of inoculation stem section in each inoculation bottle, cultivation results is specifically as shown in table 1.
The first culture base of table 1 different model to inoculation stem section at the beginning of the cultivation results of culture
Figure BDA00002734768800071
In the table 1 just the result of the test of culture show that so that just the culture base effect of cultivating is best in first 6 groups, not only bud induces survival rate up to 93.3%, and the axillalry bud growing way is better, shown in Fig. 1 a, 1b; Secondly induce survival rate also to reach 76.7% for first 5 groups of medium buds, the axillalry bud growing way is better; Although first 4 groups of medium buds induce survival rate also to reach 73.3%, the axillalry bud growing way is general.Although first 1 group, first 2 groups of medium buds induce survival rate higher, the axillalry bud growing way is bad, first 3 groups of culture medium inoculateds failure.
Operating 3 subcultures cultivates
5 leaves, 1 terminal bud that obtains take first culture, the aseptic sprouting of length about 1.2cm are seeded to respectively in the inoculation bottle of the subculture medium that is provided with different model as shown in table 2 as material, and label continues 1~continues 11 groups, and 10 bottles every group, 3 of every bottle grafts.
Observe the cultivation situation in each inoculation bottle, the result is specifically as shown in table 2.
The subculture medium of table 2 different model carries out the cultivation results that subculture is cultivated
Figure BDA00002734768800081
By the result of record in the table 2 as can be known, in 11 subculture cultivation groups, 6 groups, continue 7 groups, continue 8 groups, 9 groups the subculture medium of continuing are cultivated as best, shown in Fig. 2 a, 2b strong sprout of aseptic seedling, propagation to continue; When 6-BA2.0~3.0mg/L+IAA0.07~0.1mg/L, growth coefficient is all more than 5.0, the Multiple Buds robust growth.And two processing methods of five processing methods of 6-BA0.6~1.5mg/L+IAA0.06~1.0mg/L and 6-BA2.0~3.0mg/L+IAA1.5mg/L, its growth coefficient all is lower than 5.0.
Operate 4 culture of rootage
The Multiple Buds that shoot proliferation cultivate is obtained is cut into the stem section about 1.5cm, be inoculated in the inoculation bottle of the root media that contains different model as shown in table 3 and carry out culture of rootage, numbering is given birth to 1~living 3 groups, observes the cultivation situation in each inoculation bottle, and cultivation results is specifically as shown in table 3.
As shown in Table 3, through the cultivation of taking root in 10 days, group training seedling began to have the former base of root to occur, after 30 days, giving birth in 1 group of medium 243 strains inoculation aseptic seedling has 222 strains to take root, and rooting rate reaches 91.4%, 707 of total root systems, 3.18 root systems of average every strain, the longest root system reaches 2.8cm, shown in Fig. 3 a, 3b.Giving birth in 2 groups of medium 50 strains inoculation aseptic seedling has 40 strains to take root, and rooting rate is 80%.Giving birth in 3 groups of medium 60 strains inoculation aseptic seedling has 36 strains to take root, and rooting rate is 60%.Therefore, can see 1 group of used medium culture best results of birth.
The different root medias of table 3 carry out the result of culture of rootage
Figure BDA00002734768800091
Operating 5 hardenings processes
The oil tea group training seedling that has complete root system after the culture of rootage is taken out from inoculation bottle, with running water the root medium is rinsed well, be transplanted to respectively to be equipped with in the plastic basin of 3 kinds of different substrates that yellow soil adds peat, light substance Nutrition Soil and multiple types Nutrition Soil and cultivate, matrix is with 1: 2500 solution disinfection of potassium permanganate.Observe and transplant rear group training seedling upgrowth situation.
Yellow soil adds the peat growing medium preparation: 48~52% yellow soils+48~52% northeast peat, pH3.6.It is husky ventilative to put 1/4 clear water for the bottom of the little plastic basin of transplanting hardening, the upper strata is filled to 3/4 of basin with the Nutrition Soil of preparation, hardening 30 strains, irrigate normal root water (thimerosal of preparation) after the cultivation, cover upper film moisturizing (about humidity 76%), place 25~28 ℃ of greenhouses to cultivate, light application time 10h, intensity of illumination 1000~1500LX.
Light substance Nutrition Soil substrate preparation: 48~52% yellow soils+28~32% northeast peat+9~11% perlites+9~11% vermiculites, slow fertilising 3kg/m 3, pH4.7.It is husky ventilative to put 1/4 clear water for the bottom of the little plastic basin of transplanting hardening, the upper strata is filled to 3/4 of basin with the Nutrition Soil of preparation, hardening 30 strains, irrigate normal root water (thimerosal of preparation) after the cultivation, cover upper film moisturizing (about humidity 76%), place 25~28 ℃ of greenhouses to cultivate, light application time 10h, intensity of illumination 1000~1500LX.
Multiple types Nutrition Soil substrate preparation: 28~32% sawdust (or rice husk), 23~27% yellow soils, 4~6% ash, 28~32% northeast peat, 4~6% perlites, 4~6% vermiculites, slow fertilising 3kg/m 3, pH4.86.It is husky ventilative to put 1/4 clear water for the bottom of the little plastic basin of transplanting hardening, the upper strata is filled to 3/4 of basin with the mesh bag Nutrition Soil of preparation, hardening 240 strains, irrigate normal root water (thimerosal of preparation) after the cultivation, cover upper film moisturizing (about humidity 76%), place 25~28 ℃ of greenhouses to cultivate, light application time 10h, intensity of illumination 1000~1500LX.
The result:
(a) transplant after the 3rd day, add the group training seedling that peat transplants as matrix take yellow soil and substantially all fall leaf, remaining top only, group training seedling was all dead in 3~5 days, and the group training seedling growing way of transplanting take light substance Nutrition Soil and multiple types Nutrition Soil as matrix is better.
(b) transplant rear two weeks, the group of transplanting take the light substance Nutrition Soil as matrix training seedling growing way is general,, therefore waters once maintenance Nutrition Soil moisture about a week because this Nutrition Soil moisture content easily evaporates.The group training seedling of transplanting take the multiple types Nutrition Soil as matrix grows fine, and Nutrition Soil water conservation performance of keeping humidity is good.
(c) transplanted rear one month, the part group training seedling of transplanting with the light substance Nutrition Soil begins to send out young leaves, and growing way is slower, and storage rate is more than 85%; The group training seedling of transplanting with the multiple types Nutrition Soil substantially all grows 1~3 young leaves, robust growth, and through one month cultivation, storage rate was up to 96.9%, shown in Fig. 4 a, 4b, 4c again.After this can transfer to the land for growing field crops hardening, need the moisture-heat preservation processing of shading.

Claims (7)

1. subculture medium is used in oil tea group training seedling breeding, it is characterized in that: comprise MS+6-BA0.6~3.0mg/L+IAA0.06~1.5mg/L.
2. an oil tea is organized the propagation method of training seedling, it is characterized in that: comprise that the inoculation stem section that intercepting sterilization on the oil tea plant is obtained is carried out first culture, subculture is cultivated and the aseptic seedling after the subculture cultivation is carried out culture of rootage, it is MS+6-BA0.6~3.0mg/L+IAA0.06~1.5mg/L that described subculture is cultivated used subculture medium.
3. oil tea group as claimed in claim 1 is trained the propagation method of seedling, and it is characterized in that: the used root media of described culture of rootage is 1/2~1/8MS+IBA2.4~4.8mg/L+NAA2.2~4.6mg/L.
4. oil tea group as claimed in claim 2 is trained the propagation method of seedling, it is characterized in that: the described just used first culture base of culture is MS+6-BA0.6~1.0mg/L+IAA0.07~1.3mg/L.
5. oil tea group as claimed in claim 2 is trained the propagation method of seedling, it is characterized in that: this propagation method also comprise will obtain after the culture of rootage group training transplantation of seedlings carry out hardening to the hardening matrix and process, hardening matrix is mixed by 28~32 weight portion sawdust/rice husks, 23~27 weight portion yellow soils, 4~6 weight portion ash, 28~32 weight portion northeast peat, 4~6 weight portion perlites and 4~6 weight portion vermiculites and is consisted of.
6. organize the propagation method of training seedlings such as claim 2 or 3 or 4 or 5 described oil teas, it is characterized in that: described inoculation stem section is the inoculation branch of the robust growth that intercepts from tea oil tree, every blade on the inoculation branch is clipped 3/4~5/6 carry out clipping again remaining blade and an inoculation branch base portion behind the cleaning and sterilizing, then intercept by an axillalry bud one stem section and obtain.
7. such as the propagation method of the described oil tea group of any one in the claim 2~6 training seedling, it is characterized in that concrete operating procedure is:
First culture: the inoculation stem section after will sterilizing is seeded in the inoculation bottle that is provided with first culture base, and inoculation bottle tightened with sealed membrane sealing, again the inoculation bottle lucifuge was processed 4~6 days, then carried out illumination cultivation 30~40 days, first culture base is MS+6-BA0.6~1.0mg/L+IAA0.07~1.3mg/L;
Subculture is cultivated: the aseptic seedling that first culture is obtained is transferred to carries out strong sprout and propagation is cultivated simultaneously in the inoculation bottle that is provided with subculture medium, cultivated 35~45 days, and subculture medium is MS+6-BA0.6~3.0mg/L+IAA0.06~1.5mg/L;
Culture of rootage: subculture is cultivated the Multiple Buds obtain be cut into 1.2~2.0cm and contain respectively the aseptic seedling of tender shoots and be seeded to cultivate in the inoculation bottle that is provided with root media and take root, cultivated 25~35 days, root media is 1/2~1/8MS+IBA2.4~4.8mg/L+NAA2.2~4.6mg/L.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103461143A (en) * 2013-09-30 2013-12-25 中南林业科技大学 Method for tissue culture and rapid propagation of camellia oleifera
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CN113767850A (en) * 2021-10-15 2021-12-10 上海培林生物科技有限公司 Method for promoting proliferation and rooting of camellia oleifera and application thereof
CN115589943A (en) * 2022-08-12 2023-01-13 安徽农业大学(Cn) In-vitro regeneration method of tea trees

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CN103461143A (en) * 2013-09-30 2013-12-25 中南林业科技大学 Method for tissue culture and rapid propagation of camellia oleifera
CN106069349A (en) * 2016-08-03 2016-11-09 张琦 A kind of oil tea cutting seedling transplants Light medium
CN113767850A (en) * 2021-10-15 2021-12-10 上海培林生物科技有限公司 Method for promoting proliferation and rooting of camellia oleifera and application thereof
CN115589943A (en) * 2022-08-12 2023-01-13 安徽农业大学(Cn) In-vitro regeneration method of tea trees
CN115589943B (en) * 2022-08-12 2023-12-05 安徽农业大学 In-vitro regeneration method for tea trees

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