CN101822160B - Method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting - Google Patents

Method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting Download PDF

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CN101822160B
CN101822160B CN2010101631188A CN201010163118A CN101822160B CN 101822160 B CN101822160 B CN 101822160B CN 2010101631188 A CN2010101631188 A CN 2010101631188A CN 201010163118 A CN201010163118 A CN 201010163118A CN 101822160 B CN101822160 B CN 101822160B
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grafting
oil
stock
seed
bud
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CN101822160A (en
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蔡玲
王以红
黄金使
韦颖文
吴幼媚
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广西壮族自治区林业科学研究院
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/22Improving land use; Improving water use or availability; Controlling erosion
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/40Afforestation or reforestation

Abstract

The invention discloses a method using tissue-subculture sprouts of oil-tea camellia (Camellia oleifera Abel) to raise seedlings by grafting, particularly a method using the tissue-subculture sprout of high-grade oil-tea camellia as a scion and using the germinated germ of oil-tea camellia seeds as a stock to culture a novel plant in a shed sealed by plastic film at 10 to 35 DEG C with the air humidity being 100% on the basis of the method of cleft grafting or slope grafting, wherein the stock seeds are stored at 4 to 5 DEG C by humid sand in an icehouse having high grafting capacity and seed storage capacity; and the grafting process is carried out day by day, and the germination acceleration is carried out group by group, accordingly, the survival rate reaches 75% to 95%. The method of the invention has the advantages of high reproductive rate, high seedling raising capacity, simple method, easy operation and low material consumption; and meanwhile, the offspring can maintain the good hereditary property of the stock plant, thus solving the problem that a large amount of improved branches and budlings are required for the existing method for the vegetative propagation of oil-tea camellia, such as cuttage propagation and seedling-raising by germ-stock grafting, and meeting the requirements of high-grade and high-quality clonal oil-tea camellia sprouts for the development of the oil-tea camellia industry in China.

Description

Method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting
Technical field:
The invention belongs to plant Fast Asexual Propagation Technique field, relate to a kind of method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting.
Background technology:
Oil tea (Camellia oleifera Abel) is Theaceae (Theacae) Camellia (Camellia) plant, oil tea seed oil content height, be the distinctive edible oil materials seeds of China, with olive, oil palm, coconut also claims the world four big edible woody oil tree species, and with the tea oil that its seed extracts, oleic acid and linoleic acid are that main unsaturated fatty acid content accounts for more than 90%, compare favourably with olive oil, be described as " east olive oil ".Do not have cholesterol in the tea oil, the cholesterol in the human serum is increased, be difficult for causing that human vas sclerosis and blood pressure increase, and storage endurance, be difficult for becoming sour, can not produce the aflatoxins that causes the human carcinogen.In recent years international food and agricultural organization classifies the health type edible oil that emphasis is promoted as with the wild tea oil of China.The oil tea main producing region of China concentrates to be distributed in 17 provinces, autonomous regions and municipalities such as Guangxi, Hunan, Jiangxi, Zhejiang, Fujian, Guangdong, Hubei, Guizhou, Anhui, Yunnan, Chongqing, Henan, Sichuan and Shaanxi.Have the camellia oleifera lam area at present these oil tea main products and surpass 4,500 ten thousand mu, only Guangxi just has 6,000,000 mu." the national camellia oleiferaindustry development plan (2009-2020) " that three departments such as National Development and Reform Committee print and distribute, proposition will strive making China's oil tea plantation total scale to reach 7,000 ten thousand mu.The Eleventh Five-Year Plan period, the State Administration of Forestry utilizes the comprehensive agricultural development project funds, gives special assistance to and builds 50 high-yield tea-oil demonstration bases.
Traditional oil tea propagation technique directly is sowed at the seedbed with the oil tea seed, cultivates 1~2 year, goes out the garden with bareroot seedling, and this is the conventional seedbed system method.Its weak point is: 1, forest is very different, yields poorly, only 3~4 kilograms of every per mu yield tea oil; 2, bear fruit late, transplant afforestation after, needed just can bear fruit in 8 years, above could the results of 10 noons just can enter high yield period more than 15 years; 3, a little less than the property of Hangzhoupro, mainly contain anthracnose, soft rot and bituminous coal disease etc.; 4, breeding nursery stock difficulty adopts seed sowing reproductive survival rate only about 20%.
The asexual reproduction method of oil tea mainly contains and connect cottage propagation, inserts that leaf is grown seedlings, sprout stock is transferred three kinds: cottage propagation is a method the most simple and easy to do in the vegetative propagation, be that to choose kind good, sturdy perfectly straight, axillalry bud sound, blade life in complete 1 year is done plugged ear and grown seedlings with interior the spring tip, summer shoot; It then is the healthy and strong elite stand of selecting life in 5~15 years that oil tea inserts the leaf seedling growing process, pluck the blade at its outer rim tree crown 1/3rd places that face south, the spring inserts and will pluck the Lao Ye that passes the winter, and will select the base portion leaf of giving birth to then in two seasons of autumn in summer, it is to the south that spring and autumn is inserted the blade face, northwards grows seedlings in the blade face that the summer inserts; Oil-tea sprout anvil graft technology is to select common oil tea large seed, transfers anvil at indoor sand bed Pregermination and seedling breeding, selects for use the full oil tea bud of the current-year branch that growth is sturdy, axillalry bud is obvious, anosis worm endangers to do scion, a fringe one bud and the grafting of seedling anvil.
The cloning technology for high-yield tea-oil tree of Chinese patent CN1258434A.Be in existing oil tea colony woods, to survey in continuous 3 years to produce to select excellent back cloneization to set up select tree to collect the garden, produce through observation in 1~2 year and survey in continuous 4 years, select the high select tree of tea fruit output to set up high yield clone and adopt firmly, gather then branch cut into single tea tree germ sow be connected on the bud seedling that the deep-fried twisted dough sticks seed cultivates cultivation 2 years high yield oil tea sapling.
A kind of rapid propagation method for clonal tissue culture of oil tea of Chinese patent CN 101530063A.Concrete operations are as follows: (1) is transferred to the aseptic seedling that obtains on the subculture medium and is bred, the medium wide-mouth bottle packing of 300ml, and every bottle of 30ml seals the film wrapping, at 121 ℃ and 1.1kg.cm 2Under the condition, with the high-pressure steam sterilizing pan 20min that sterilizes.The screening of condition of culture: in cultivation temperature is 2 ℃ in 25 soil, illumination 1500~2000Lx, and every day, light application time was to cultivate in the culturing room of 12h.(2) will breed the aseptic seedling that obtains and transfer on the strong seedling culture base and carry out strong sprout, medium is with the wide-mouth bottle packing of 300ml, and every bottle of 30ml seals the film wrapping, at 121 ℃ and 1.1kg.cm 2Under the condition, with the high-pressure steam sterilizing pan 20min that sterilizes.Condition of culture is with (1).(3) aseptic seedling is taken out from blake bottle, clean the root medium, and its base portion soaked in KIBA solution, in disposal plastic cup, a cup mesostroma is prepared by a certain percentage by yellow soil, charing rice husk ash, fine sand with the nursery stock cuttage, the rim of a cup use is sealed the film covering and is preserved moisture, cup is placed the management of taking root of the green house of tool intermittent spraying facility, and temperature keeps 20~30 ℃, 10 minutes intermittent times of spraying, spray time 10 seconds, rooting rate 87%.It is characterized in that: the formation of plant is on same individuality, induces plant organ's polarity in differentiation and forming by exogenous hormone.Promptly utilize the bud organ aseptic, artificially control under nutrition, temperature and the intensity of illumination condition, add certain exogenous hormone, root induction forms.
The propagation method of the using oil-tea young anvil to graft containerized seedlings of Chinese patent CN 101611688A.Propagation method is operated according to the following steps: (1) cultivates young anvil, and concrete operations are as follows: 1. garden ground is selected: select to have water source or pouring condition and the field is done in the place being convenient to manage; 2. carry out the seedbed and carry out container and put: with ground, garden smooth and divided seedbed and pavement after, the container that nutrition soil will be housed again is placed on the seedbed with neatly being close to; 2. seed treatment: educate seed selection, sterilization and the vernalization of anvil seed before the sowing, seed is selected the common oil tea seed in the oil tea species for use: 4. anvil is educated in sowing: sow in container annual February December to next year after the presprouting of seeds; 5. stock management: after planting regularly trickle is preserved moisture, and treats to carry out after nursery stock is unearthed the control of rich water quality management and damage by disease and insect; (2) propagation by grafiting, concrete operations are as follows: 1. build cool canopy: the nursery lot of cultivating stock is put up cool canopy respectively to shade before grafting; 2. grafting: the seedling with 4-6 month is a stock, adopts semi-lignified to lignified Camellia Oleifera Clones branch to do scion, carries out grafting in the suitable phenophase of deep-fried twisted dough sticks; 2. preserve moisture: after the grafting immediately arch cover preserve moisture, humidity remains on 80% one 90% in the cover; 4. managing after grafting: carry out in time remove sprout, the control of rich water quality management and damage by disease and insect.
By the above as can be known, traditional oil tea propagation technique is with the direct seeding and seedling raising of oil tea seed.Because oil tea is a cross-pollinatd plant, it is bigger to make a variation in long-term sexual reproduction process, and good and bad difference great disparity is difficult to keep the merit of maternal plant, and general 6~8 years beginnings were the one of the main reasons that causes the oil tea low yield really.And the vegetative method of existing oil tea, as connect cottage propagation, insert growing seedlings that leaf is grown seedlings and sprout stock is transferred, need a large amount of breeding budlings, limited because of the budling of breeding, be difficult to satisfy the demand of national camellia oleiferaindustry development to Camellia Oleifera Clones.Need the propagation technique deficiency of a large amount of breeding budlings in order to overcome the of poor quality and existing asexual reproduction method of seed sowing, of the present invention group of training subculture bud grafting seedlings-growing method is the good method of oil tea matter breeding seedling fast asexual propagation.This method has that reproduction speed is fast, method is simple, processing ease, material consumption is few, be not subjected to the characteristics of season and region restriction, can produce the higher group training subculture bud grafting fringe of genetic stability in a short time in batches, filial generation simultaneously can keep the good hereditary capacity of maternal plant.
Summary of the invention:
Purpose of the present invention: in order to solve above-mentioned problems of the prior art, the efficient group culturation rapid propagating technology of exploitation Camellia Oleifera Clones aims to provide a kind of method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting.
Technical scheme of the present invention is: have the callus ability after utilizing plant injured, the mechanism that tender tissue callus ability is stronger, oil tea group training bud with high-quality is scion, the plumule that high quality seed is sprouted is a stock, adopt the grafting method of cleft grafting or mitered, utilize tinfoil paper colligation interface, the cortex of scion and stock is healed closely form the technology of whole plant.
The present invention is achieved in that
Method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting comprises seed selection, vernalization, cultivation, grafting, grows seedlings and goes out the garden operational sequence, and the concrete operations step is as follows:
(1) seed selection: select no damage by disease and insect, high quality seed that particle is big and full for use;
(2) vernalization: after seed is sterile-processed, uses the sand breeding, measure greatly if desired, can adopt icebox to refrigerate vernalization in batches;
(3) cultivate: with the oil tea of the high-quality organ of sprouting then is explant, and aseptic bud is obtained in the sterilization inoculation, aseptic bud is transferred to bred on the subculture medium and strong seedling culture;
(4) grafting: adopting high-quality oil tea group training subculture bud is scion, is stock with the plumule of seed germination, takes cleft grafting method or the grafting of mitered method;
(5) grow seedlings: build the high shading shed of 180cm, degree of shading 75% is set up the little shed of the high plastics diaphragm seal of 60cm the oil tea grafting is cultivated under the cool canopy;
(6) go out the garden: scion is grown can go out garden afforestation more than the high 15cm.
The above-described process that refrigerates vernalization in batches with sand breeding, icebox, be to fill up the thick wet sand of one deck 14~16cm on the ground, evenly sprinkle one deck seed, it is thick to cover husky 3cm again, the regular vernalization of preserving moisture of spraying water, the grafting number is big, and storage kind of amount is many, and grafting is carried out day by day, seed is taked vernalization in batches with profit sand stored under refrigeration in 4~5 ℃ of iceboxs.
Above-described aseptic bud being transferred to the process of breeding on the subculture medium with strong seedling culture, is that every bottle of 60~80ml seals with cotton balls with the triangular flask packing of medium with 500ml, again with brown paper wrapping cotton balls and bottleneck, at 120 ℃ and 1.1kg.cm 2Under the condition, with the high-pressure steam sterilizing pan 15min that sterilizes, culturing room adopts the illumination of the natural scattering light of 1000~5000Lx.
The above-described process of taking cleft grafting method or the grafting of mitered method, be with successive transfer culture 30~40d, long 1.5~the 7cm of bud, contain 2~10 in blade, the strong bud that axillalry bud is 1~10 is cut with moistening gauze clad, the scion base portion being whittled into the inclined-plane of two symmetries or cutting an inclined-plane from left to right with scalpel becomes long wedge shape of 0.4~1.0cm or inclined-plane at 45, take cleft grafting method or mitered method, high 3~10cm, the seed plumule that contains 2~8 of true leaves cuts, group training subculture bud is inoculated on the plumule of seed germination,, cultivates and form whole plant with tinfoil paper colligation grafting mouth.
The process that above-described oil tea grafting is cultivated is 10~35 ℃ of the interior temperature of control plastic film sealing canopy, air humidity 100%, in preceding 10d, open plastic film 1 time every 2d, the time is 5~30min, open every day 1 time, the time is 10~90min, in time pours water backward, moisture does not touch interface, in time removes the rudiment of stock, and grafting 40~50d can open film, remove tinfoil paper during 60~80d, suitable sealing fertilizer, concentration 0.05~3%, look the growth of nursery stock, from thin to thick.
Above-described cleft grafting method with scalpel excision stock terminal bud, is split a cutter at the center of stock from top to bottom, and the degree of depth is 0.6~1cm, pries open stock and splits mouth, and stock is inserted in scion from top to bottom, uses the tinfoil paper colligation tight at last.
Above-described mitered method, with scalpel excision stock terminal bud, right-to-left is cut 1 scarf at 45 to stock, and the edge alignment of scion inclined-plane and stock inclined-plane bud is joined, and uses the tinfoil paper colligation tight at last.
Advantage of the present invention and good effect:
1, method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting of the present invention.With high-quality oil tea group training subculture bud is scion, the plumule of seed germination is a stock, by cleft grafting or mitered grafting method, the cambium of group training subculture bud and the cambium of stock are combined closely, utilize tinfoil paper colligation interface, 10~35 ℃ of temperature, air humidity 100%, cultivate the whole plant that makes new advances in the canopy of plastic film sealing, can produce the higher group training subculture bud grafting of genetic stability in a short time in batches, filial generation can keep the good hereditary capacity of maternal plant.Solved existing oil tea asexual reproduction method such as cottage propagation, the required a large amount of breeding branch of sprout stock graft seedling growth and breeding budling problem simultaneously, can satisfy the demand of national camellia oleiferaindustry development Camellia Oleifera Clones good quality and high output oil tea seedling.
2, the stock seed stores in 4~5 ℃ of iceboxs with profit is husky, and it is big to adapt to the grafting number, and storage kind of amount is many, and grafting is carried out day by day, takes vernalization in batches.Adopt this method grafting, survival rate is 75%~95%.Have that reproduction speed is fast, the amount of growing seedlings is big, method is simple, processing ease, material consumption is few, is not subjected to the characteristics of season and region restriction, can adapt to the needs of China's camellia oleiferaindustry development.
Embodiment:
Below in conjunction with embodiment, method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting of the present invention is further described.
Embodiment 1
Grafting time is April, when oil tea is gathered, selects local adaptation strong, and well-grown oil tea provenance seed is made stock.Select the big grain full seed of no damage by disease and insect, sterile-processed after, fill up the thick wet sand of one deck 15cm on the ground, evenly sprinkle one deck seed, it is thick to cover husky 3cm again, the regular vernalization of preserving moisture of spraying water when radicle is about 2cm, moves to seed the interior or upward cultivation of bed of growing seedlings of container of 9cm * 13cm.The grafting number is big, and storage kind of amount is many, and grafting is carried out day by day, and seed stores in 4 ℃ of iceboxs with profit is husky, takes vernalization in batches.Oil tea group training subculture bud organ with high-quality is an explant, and aseptic bud is obtained in sterilization, aseptic bud is transferred to bred on the subculture medium and strong seedling culture.The medium triangular flask packing of 500ml, every bottle of 70ml seals with cotton balls, again with brown paper wrapping cotton balls and bottleneck, at 120 ℃ and 1.1kg.cm 2Under the condition, with the high-pressure steam sterilizing pan 15min that sterilizes, culturing room adopts the illumination of the natural scattering light of 1500~4000Lx.
With oil tea group training subculture bud is scion, and the oil tea plumule is a stock.Adopt cleft grafting method to carry out grafting.Scion is for cultivating the group training subculture bud of 35d, and the long 2~6cm of bud contains 2~10 in blade, 2~10 of axillalry buds, and the shearing back is stand-by with moistening gauze clad; Stock is the plumule that full seed is sprouted, and high 6~10cm contains 5~8 of true leaves.With scalpel the scion base portion is whittled into the inclined-plane of two symmetries, makes it to become the long wedge shape of 0.5cm.Require straight, smooth, the cleaning of tangent plane, the branch skin can not perk.With scalpel excision stock terminal bud, stay a blade.Firmly split a cutter at the center of stock from top to bottom, the degree of depth is 0.6~1cm, pry open stock and split mouth, stock is inserted in scion from top to bottom, attention is alignd the cortex (cambium) of scion and stock and it is coincide closely, use the tinfoil paper colligation tight at last, place the top of plastic greenhouse to build the high shade of 180cm, set up under the cool canopy of shade density 75% in the little shed of the high plastics diaphragm seal of 60cm, keep 100% air humidity, cultivate 40~44d, the interface healing of scion and stock forms whole plant.10d opens plastic film 1 time every 2d before after the grafting, and time 8~10min opens film every day 1 time backward, and time 40~60min in time pours water, and keeps 20~30 ℃ of temperature in the plastic greenhouse, 100% air humidity, and moisture does not touch interface.In time remove the rudiment of stock.Grafting 15~20d fringe bud begins to sprout, and the growth of 23~28d fringe bud young leaves shows that scion and stock otch heal, and opens film this moment, notes keeping nutrient cup or medium of seedling bed moistening.Grafting 70~72d, the nursery stock wound heals fully, enters normal growth, can remove tinfoil paper, suitable sealing fertilizer, concentration 0.05~3% is looked the growth of nursery stock, from thin to thick.The fringe blastogenesis is grown tall 15cm when above, can go out garden afforestation.Graft survival rate reaches 95%.
Embodiment 2
Grafting time is February, when oil tea is gathered, selects local adaptation strong, and well-grown oil tea provenance seed is made stock.Select the big grain full seed of no damage by disease and insect, sterile-processed after, fill up the wet sand of about 15 cm thicks of one deck on the ground, evenly sprinkle one deck seed, it is thick to cover husky 3cm again, the regular vernalization of preserving moisture of spraying water when radicle is about 2cm, moves to seed the interior or upward cultivation of bed of growing seedlings of container of 9cm * 13cm.The grafting number is big, and storage kind of amount is many, and grafting is carried out day by day, and seed stores in 4.5 ℃ of iceboxs with profit is husky, takes vernalization in batches.With the oil tea group training successive transfer culture of high-quality, the bud organ is an explant, and aseptic bud is obtained in sterilization, aseptic bud is transferred to bred on the subculture medium and strong seedling culture.The medium triangular flask packing of 500ml, every bottle of 70ml seals with cotton balls, again with brown paper wrapping cotton balls and bottleneck, at 120 ℃ and 1.0kg.cm 2Under the condition, with the high-pressure steam sterilizing pan 15min that sterilizes, culturing room adopts the illumination of the natural scattering light of 1000~3000Lx.
With oil tea group training subculture bud is scion, and the oil tea plumule is a stock.Adopt cleft grafting and mitered method to carry out grafting.Scion is for cultivating the group training subculture bud of 40d, and the long 1~5cm of bud contains 1~8 in blade, 1~8 of axillalry bud, and the shearing back is stand-by with moistening gauze clad; Stock is the plumule that full seed is sprouted, and high 5~8cm contains 4~6 of true leaves.With scalpel the scion base portion is whittled into and cuts an inclined-plane from left to right, make it to become 45 ° long inclined-plane of 0.4~0.5cm.Tangent plane is straight, smooth, cleaning, and the branch skin can not perk.With scalpel excision stock terminal bud, stay a blade.The mitered method: firmly right-to-left is cut 1 scarf at 45 to stock, scion inclined-plane and stock inclined-plane join, attention is alignd the cortex (cambium) of scion and stock and it is coincide closely, use the tinfoil paper colligation tight at last, place the top of plastic greenhouse to build the high shade of 180cm, set up under the cool canopy of shade density 75% in the little shed of the high plastics diaphragm seal of 60cm, keep 100% air humidity, cultivate 42~46d, the interface healing of scion and stock forms whole plant.10d opens plastic film 1 time every 2d before after the grafting, and time 6~8min opens film every day 1 time backward, and time 30~50min in time pours water, and keeps 10~25 ℃ of temperature in the plastic greenhouse, 100% air humidity, and moisture does not touch interface.In time remove the rudiment of stock.Under the normal condition, grafting 30~35d fringe bud begins to sprout, and the growth of 45~50d fringe bud young leaves shows that scion and stock otch heal, and opens film this moment, notes keeping nutrient cup or medium of seedling bed moistening.Grafting 75~80d, the nursery stock wound heals fully, enters normal growth, removes tinfoil paper, suitable sealing fertilizer, concentration 0.05~3% is looked the growth of nursery stock, from thin to thick.The fringe blastogenesis is grown tall 15cm when above, can go out garden afforestation.Graft survival rate 92%.
Embodiment 3
Grafting time is June, when oil tea is gathered, selects local adaptation strong, and well-grown oil tea provenance seed is made stock.Select the big grain full seed of no damage by disease and insect, sterile-processed after, fill up the thick wet sand of one deck 16cm on the ground, evenly sprinkle one deck seed, it is thick to cover husky 3cm again, the regular vernalization of preserving moisture of spraying water when radicle is about 2cm, moves to seed the interior or upward cultivation of bed of growing seedlings of container of 9cm * 13cm.The grafting number is big, and storage kind of amount is many, and grafting is carried out day by day, and seed stores in 4~5 ℃ of iceboxs with profit is husky, takes vernalization in batches.With the oil tea group training successive transfer culture of high-quality, the bud organ is an explant, and aseptic bud is obtained in sterilization, aseptic bud is transferred to bred on the subculture medium and strong seedling culture.The medium triangular flask packing of 500ml, every bottle of 70ml seals with cotton balls, again with brown paper wrapping cotton balls and bottleneck, under 120 ℃ and 1.1kg.cm2 condition, with the high-pressure steam sterilizing pan 15min that sterilizes, culturing room adopts the illumination of the natural daylight of 2000~5000Lx.
With oil tea group training subculture bud is scion, and the oil tea plumule is a stock.Adopt cleft grafting and mitered method to carry out grafting.Scion is for cultivating the group training subculture bud of 32d, and the long 3~7cm of bud contains 3~8 in blade, 2~6 of axillalry buds, and the shearing back is stand-by with moistening gauze clad; Stock is the plumule that full seed is sprouted, and high 3~6cm contains 2~5 of true leaves.With scalpel the scion base portion is whittled into the inclined-plane of two symmetries or cuts an inclined-plane from left to right, make it to become the long wedge shape of 0.4cm.Tangent plane is straight, smooth, cleaning, and the branch skin can not perk.With scalpel excision stock terminal bud, stay a blade.Cleft graft: firmly split a cutter at the center of stock from top to bottom, the degree of depth is 0.6~0.8cm, pry open stock and split mouth, stock is inserted in scion from top to bottom, attention is alignd the cortex (cambium) of scion and stock and it is coincide closely, use the tinfoil paper colligation tight at last, place the top of plastic greenhouse to build the high shade of 180cm, in the canopy of the plastic film sealing of shade density 75%, keep 100% air humidity, cultivate 40~50d, the interface healing of scion and stock forms whole plant.10d before after the grafting opens plastic film 1 time every 1d, and time 20~30min opens film every day 1 time backward, and time 60~90min in time pours water, and keeps 20~35 ℃ of temperature in the plastic greenhouse, 100% air humidity, and moisture does not touch interface.In time remove the rudiment of stock.Under the normal condition, grafting 15~20d fringe bud begins to sprout, and the growth of 25~30d fringe bud young leaves shows that scion and stock otch heal, and opens film this moment, notes keeping nutrient cup or medium of seedling bed moistening.Grafting 60d nursery stock wound heals fully, enters normal growth, removes tinfoil paper, suitable sealing fertilizer, and concentration 0.05~3% is looked the growth of nursery stock, from thin to thick.The fringe blastogenesis is grown tall 15cm when above, can go out garden afforestation.Graft survival rate 75%.

Claims (2)

1. a method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting comprises seed selection, vernalization, cultivation, grafting, grows seedlings and goes out the garden operational sequence, and it is characterized in that: the concrete operations step is as follows:
(1) seed selection: select no damage by disease and insect, high quality seed that particle is big and full for use;
(2) vernalization: after seed is sterile-processed, refrigerate vernalization in batches with sand breeding, icebox, be to fill up the thick wet sand of one deck 14~16cm on the ground, evenly sprinkle one deck seed, it is thick to cover husky 3cm again, the vernalization of preserving moisture of regularly spraying water, and the grafting number is big, storage kind of amount is many, grafting is carried out day by day, and seed is taked vernalization in batches with profit sand stored under refrigeration in 4~5 ℃ of iceboxs;
(3) cultivate: with the oil tea of the high-quality organ of sprouting then is explant, aseptic bud is obtained in the sterilization inoculation, aseptic bud transferred to breed on the subculture medium and strong seedling culture, be with the triangular flask packing of medium with 500ml, every bottle of 60~80ml, seal with cotton balls, again with brown paper wrapping cotton balls and bottleneck, at 120 ℃ and 1.1kg.cm 2Under the condition, with the high-pressure steam sterilizing pan 15min that sterilizes, culturing room adopts the illumination of the natural scattering light of 1000~5000Lx;
(4) grafting: adopting high-quality oil tea group training subculture bud is scion, plumule with seed germination is a stock, take cleft grafting method or the grafting of mitered method, be with successive transfer culture 30~40d, long 1.5~the 7cm of bud, contain 2~10 in blade, the strong bud that axillalry bud is 1~10 is cut with moistening gauze clad, the scion base portion being whittled into the inclined-plane of two symmetries or cutting an inclined-plane from left to right with scalpel becomes long wedge shape of 0.4~1.0cm or inclined-plane at 45, take cleft grafting method or mitered method, high 3~10cm, the seed plumule that contains 2~8 of true leaves cuts, group training subculture bud is inoculated on the plumule of seed germination,, cultivates and form whole plant with tinfoil paper colligation grafting mouth;
(5) grow seedlings: build the high shading shed of 180cm, degree of shading 75% is set up the little shed of the high plastics diaphragm seal of 60cm the oil tea grafting is cultivated under the cool canopy;
(6) go out the garden: scion is grown can go out garden afforestation more than the high 15cm.
2. method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting according to claim 1, it is characterized in that: described oil tea grafting is cultivated, be 10~35 ℃ of the interior temperature of control plastic film sealing canopy, air humidity 100% in preceding 10d, is opened plastic film 1 time every 2d, time is 5~30min, open every day 1 time, the time is 10~90min, in time pours water backward, moisture does not touch interface, in time remove the rudiment of stock, grafting 40~50d can open film, removes tinfoil paper during 60~80d, suitable sealing fertilizer, concentration 0.05~3% is looked the growth of nursery stock, from thin to thick.
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