CN103004593A - Thin-shell pecan tissue culture bud micro grafting method - Google Patents
Thin-shell pecan tissue culture bud micro grafting method Download PDFInfo
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- CN103004593A CN103004593A CN201210537557XA CN201210537557A CN103004593A CN 103004593 A CN103004593 A CN 103004593A CN 201210537557X A CN201210537557X A CN 201210537557XA CN 201210537557 A CN201210537557 A CN 201210537557A CN 103004593 A CN103004593 A CN 103004593A
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Abstract
The invention discloses a thin-shell pecan tissue culture bud micro grafting method which comprises the following steps of: 1. inducing and multiplying a scion tissue culture bud, wherein five working procedures of collecting and disinfecting an explant, preparing an adventitious bud inducing culture medium, inducing an adventitious bud, preparing an adventitious bud multiplying culture medium and performing adventitious bud multiplying cultivation; 2. cultivating a thin-shell pecan stock; 3. grafting the tissue culture bud; and 4. managing grafting and a healing stage. According to the method, the thin-shell pecan has the advantages of retaining the maternal characteristics along with early bearing, being high in inversion resistance, high in multiplying speed and the like.
Description
Technical field
The present invention relates to a kind of indefinite bud grafting with apocarya group training to the miniature grafting method of the annual seedling shoot of apocarya.
Background technology
Apocarya [Carya illinoinensis (Wangehn.) K.Koch.] has another name called pecan tree, pecan, be Juglandaceae hickory deciduous tree, originate in southern US and northern Mexico, it is in the world one of famous oil plant dry fruit tree variety, it simultaneously also is the good economic tree of fruit, wooden dual-purpose, can adapt to soil acidity or alkalinity in a big way, under natural environment, apocarya is by seminal propagation.The apocarya dry fruit can be processed into various characteristic typical local food, health products, and its kind benevolence oil content is rich in unsaturated fatty acid up to 70%～72%, contains crude protein 7.8%～9.6% and contains the necessary mineral elements of human body such as K, Ca, Mg, Na.Apocarya wooden hard, texture is beautiful, anticorrosive, of many uses in military project, boats and ships and building industry, be a kind of very good economic tree of China.
The existing upper one-hundred-year history of China's introducing and planting apocarya, Zhejiang Province began to introduce a fine variety the sixties in 19th century.Apocarya is as important southern characteristic dry fruit, be economic benefit best, be subjected to one of seeds that hill farmer welcomes most, the good ecological economy seeds that are fit to especially the Limestone Mountain cultivation have become the suitable important mountain region economic tree in province of promoting in the south China subtropical zones such as Zhejiang, Anhui, Yunnan, Fujian, Hunan, Jiangxi.At present, apocarya has become one of first-selected economic tree of south China forestation in mountain area.Yet owing to mostly introduce by seed in early days, existing operation woods mostly is seedling tree, and cultivation is fragmentary to be disperseed, kind is very different, flourish along with southern dry fruit industry restructuring and economic forest, and at present apocarya deterioration of variety separates obviously, the result is slow, yield poorly, poor quality, it is large to add the height of climbing the tree, and plucks inconvenient, as fruit production, Difficulty.Although regional larger area popularizing plantings such as Yunnan, Jiangsu, Zhejiang, Hunan, Guangxi, Sichuan, Jiangxi, Gansu, Henan, Guizhou, Hebei in recent years, but apocarya is few owing to high quality seedling quantity in cultivation, so that survival rate is low, afforestation is not grown into forest, poor growth, production cycle is long, yield per unit area is low, large, the good scion of low yield Low-efficiency forest area source less, the seedling propagation coefficient is low and the problem such as high-quality technology for vigorous seedling cultivation shortage becomes increasingly conspicuous, and has seriously restricted the rapid and healthy of apocarya industry.
Forestry circle, numerous scholars have done many new explorations and trial to the breeding of apocarya, apocarya root segment seedling growing process such as the Mr.s' such as Li Junnan apocarya simple bud side grafting technology, Chang Jun etc., the apocarya of Yang Guorong etc. is with the numerous seedling fast-propagation of seedling technology etc., and each has made useful contribution to society.Through retrieval, find so far to cultivate indefinite bud through miniature grafting by the excellent strain annotinous branch of apocarya through test tube, the relevant report of the method that breeding takes root breeds plant.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of take the apocarya annotinous branch as explant, and induce and form indefinite bud, Elongation of adventitious bud is cultivated, and with the indefinite bud grafting of the elongation miniature grafting method to the new skill of the annual seedling of apocarya.
Technical problem of the present invention solves by following technical solution:
The miniature grafting method of this apocarya group training bud carries out as follows:
One, scion group training bud is induced and is bred:
(1) explant collection and sterilization: choose healthy and strong 1 year healthy and strong shoot of life of the excellent strain of apocarya, carry out successively running water flushing 2h, on the superclean bench with 75% ethanol sterilization 3min, aseptic water washing 3～4 times, concentration is 0.525% NaClO sterilization 20min and vacuumizes, behind the aseptic water washing 5 times, blot surface moisture with sterilization filter paper, downcutting with terminal bud and axillalry bud length is that the branch section of 1～1.5cm is stand-by;
(2) preparation of adventitious bud induction culture base: WPM additional saccharose 20～30g/L, agar A 6.5～8.5g/L, 6-BA are 6-aminoadenine 0.5～2.0mg/L, and IBA is indolebutyric acid 0.1～0.5mg/L, pH value 5.7;
(3) adventitious bud inducing: in the adventitious bud induction culture base with the young shoot access aforementioned (2) that radicle growth is good behind the first culture, through inducing of 21d, obtain indefinite bud; With microcomputer time-controlled switch control culturing room light application time every day, every day, light dark period was the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
(4) preparation of adventitious bud proliferation medium: WPM additional saccharose 20～30g/L, agar A 6.5～8.5g/L, 6-BA are 6-aminoadenine 1.0～3.0mg/L, and IBA is indolebutyric acid 0.05～0.1mg/L, pH value 5.7;
(5) adventitious bud proliferation is cultivated: the indefinite bud that obtains is bred cultivation at the adventitious bud proliferation medium of aforementioned (4), and subculture was 1 time in 21 days, and subculture is 3 times altogether, and growth coefficient is 5～8; Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
Two, the apocarya stock is cultivated:
Select fully ripe then apocarya to make the stock seed, in booth (6m*30m) seedbed is set, seedbed length and width are respectively 1m and 10m, and the bottom, seedbed is by the standard cloth ground hot line of every square metre of 100W, and temperature is by the control of temperature control view; Gibberellin with 1000mg/L concentration soaked seed 2～3 days, pull out and directly seed is placed on the seedbed afterwards, the apocarya suture is vertical with the soil table, put 700～800 for general every square metre, on apocarya, cover sandy soil 8～10cm, water permeable again covering film, the control temperature is at 25 ± 2 ℃;
Three, group training bud grafting:
Take out the high seed seedling of 18～20cm from the seedbed and do stock in booth, 2～3cm cuts off at the place anvil seedling more than the cotyledon attachment region, along slightly partially rip cutting of medulla, and dark 1cm; Cut the long stalwartness group training bud of 3.0～3.5cm, insert the longitudinal incision of stock, with mocromembrane (0.5cm*10cm) wrapping, and transplant and plant in container for plant growth, at container for plant growth outer cover transparent plastic bag heat and moisture preserving; After one week, cut off gradually the bag angle of transparent plastic bag, until all cut off and remove bagging;
Four, management:
After the miniature grafting, scion and stock healing stage greenhouse temperature are controlled at 25 ± 2 ℃, and humidity is controlled at 85% ± 2%, avoids direct sunlight, until scion and stock heal fully, grafting survives.
The invention has the beneficial effects as follows:
Use apocarya group training adventitious bud inducing and organize the grafting of training bud, can make newborn plant keep maternal merit, effectively improve the reproduction coefficient of the excellent strain of apocarya, be fast numerous the laying a good foundation of apocarya breeding.Compare with seed propagation, excellent strain adventitious bud inducing can keep maternal good biological property, and the difficult problem that is difficult for taking root in the indefinite bud bottle can be effectively avoided in the grafting of excellent strain indefinite bud, and growing-seedling period is short.Comparing with cottage propagation or conventional propagation by grafiting, is cottage propagation or conventional propagation by grafiting 50～60 times at the reproduction coefficient of a growth cycle inner tissue cultivation of 6 months by a definite date.
Embodiment
The present invention is described in further detail below in conjunction with embodiment: in the scion adventitious bud inducing and proliferated culture medium of the miniature grafting method of this apocarya group training bud, selecting minimal medium is WPM medium (its composition is public), carefully do not state, this is through using MS, 1/2MS, WPM, four kinds of minimal mediums of DKW are equal additional saccharose 20～30g/L separately, agar A 6.5～8.5g/L, 6-BA is 6-aminoadenine 0.5～3.0mg/L, IBA is indolebutyric acid 0.05～0.5mg/L, pH value 5.7, test, every group is repeated 3 times, every kind of minimal medium has the comparative trial of 20 explants to draw, after processing 21 days in the WPM medium, uncertain buds growth is best, the indefinite bud quantity of inducing is maximum, best in quality.
Cultivating stock uses the gibberellin of 1000mg/L concentration to soak seed 2-3 days, the apocarya suture is vertical with the soil table, puts 700～800, cover sandy soil 8～10cm on apocarya for general every square metre, water permeable again covering film, the control temperature is at 25 ± 2 ℃;
Take out suitable seed seedling from the seedbed and do stock, 2～3cm cuts off at the place anvil seedling more than cotyledon petiole, along slightly partially rip cutting of medulla, deeply about 1cm; Treated group training bud is whittled into wedge shape, inserts the longitudinal incision of stock, with the mocromembrane wrapping, plant in container for plant growth, and the moisturizing of outer cover transparent plastic bag, shear angle removes bag in good time, carries out at last the booth management work of scion and stock healing stage.
In a word, the proportioning value of each raw material of medium disclosed in this invention also can adopt respectively replacing near value of preferred plan definite value of the present invention.
Given below is most preferred embodiment of the present invention.
This apocarya group is trained the miniature grafting method of bud, comprises two kinds of medium of different Induction periods, and raw material and the additional amount thereof of two kinds of medium are respectively:
(1) adventitious bud induction culture base: WPM additional saccharose 25g/L, agar A7.5g/L, 6-BA are 6-aminoadenine 2.0mg/L, and IBA is indolebutyric acid 0.3mg/L, pH value 5.7;
(2) adventitious bud proliferation medium: WPM additional saccharose 25g/L, agar A7.5g/L, 6-BA are 6-aminoadenine 1.0mg/L, and IBA is indolebutyric acid 0.1mg/L, and the pH value is 5.7;
Carry out apocarya adventitious bud inducing and enrichment procedure process the following step with above-mentioned medium:
At first be explant collection and sterilization: choose healthy and strong 1 year healthy and strong shoot of life of the excellent strain of apocarya, carry out successively running water flushing 2h, on the superclean bench with 75% ethanol sterilization 3min, aseptic water washing 3～4 times, concentration is 0.525% NaClO sterilization 20min and vacuumizes, behind the aseptic water washing 5 times, blot surface moisture with sterilization filter paper, (1～1.5cm) is stand-by to downcut branch section with terminal bud and axillalry bud;
Next is adventitious bud inducing: in the aforesaid just culture base of the rataria that takes out under step (1) aseptic condition access, cultivate 21d, light dark period is to cultivate intensity of illumination under the condition of the dark 8h of light 16h/: 40～50 μ mol m
-2s
-1Cultivation temperature is 25 ± 2 ℃;
Be adventitious bud proliferation at last: the indefinite bud that obtains is grown to 0.5cm left and right sides Gao Shi, downcut and be inoculated into the aforesaid adventitious bud proliferation medium from basal part of stem and cultivate, subculture was 1 time in 21 days, and subculture is 3 times altogether, and each growth coefficient is 5～8.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, 25 ± 2 ℃ of cultivation temperature.
Claims (1)
1. an apocarya group is trained the miniature grafting method of bud, it is characterized in that carrying out as follows:
One, scion group training bud is induced and is bred;
(1) explant collection and sterilization: choose healthy and strong 1 year healthy and strong shoot of life of the excellent strain of apocarya, carry out successively running water flushing 2h, on the superclean bench with 75% ethanol sterilization 3min, aseptic water washing 3～4 times, concentration is go out seedling 20min and vacuumizing of 0.525% NaClO, behind the aseptic water washing 5 times, blot surface moisture with sterilization filter paper, downcutting with terminal bud and axillalry bud length is that the branch section of 1～1.5cm is stand-by;
(2) preparation of adventitious bud induction culture base: WPM additional saccharose 20～30g/L, agar A 6.5～8.5g/L, 6-BA are 6-aminoadenine 0.5～2.0mg/L, and IBA is indolebutyric acid 0.1～0.5mg/L, pH value 5.7;
(3) adventitious bud inducing: in the adventitious bud induction culture base with the young shoot access aforementioned (2) that radicle growth is good behind the first culture, through inducing of 21d, obtain indefinite bud; With microcomputer time-controlled switch control culturing room light application time every day, every day, light dark period was the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
(4) preparation of adventitious bud proliferation medium: WPM additional saccharose 20～30g/L, agar A 6.5～8.5g/L, 6-BA are 6-aminoadenine 1.0～3.0mg/L, and IBA is indolebutyric acid 0.05～0.1mg/L, pH value 5.7;
(5) adventitious bud proliferation is cultivated: the indefinite bud that obtains is bred cultivation at the adventitious bud proliferation medium of aforementioned (4), and subculture was 1 time in 21 days, and subculture is 3 times altogether, and growth coefficient is 5～8; Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
Two, the apocarya stock is cultivated:
Select fully ripe then apocarya to make the stock seed, the seedbed is set in booth, seedbed length and width are respectively 1m and 10m, and the bottom, seedbed is by the standard cloth ground hot line of every square metre of 100W, and temperature is controlled by temperature controller; Gibberellin with 1000mg/L concentration soaked seed 2～3 days, pull out and directly seed is placed on the seedbed afterwards, the apocarya suture is vertical with the soil table, puts 700～800, covers sandy soil 8～10cm on apocarya for every square metre, water permeable again covering film, the control temperature is at 25 ± 2 ℃;
Three, group training bud grafting:
Take out the high seed seedling of 18～20cm from the seedbed and do stock in booth, 2～3cm cuts off at the place anvil seedling more than the cotyledon attachment region, along slightly partially rip cutting of medulla, and dark 1cm; Cut the long stalwartness group training bud of 3.0～3.5cm, insert the longitudinal incision of stock, with the mocromembrane wrapping of 0.5cm*10cm, and transplant and plant in container for plant growth, in the moisturizing of container for plant growth outer cover transparent plastic bag; After one week, cut off gradually the bag angle of transparent plastic bag, until all cut off and remove bagging;
Four, management: after the grafting, scion and stock healing stage greenhouse temperature are controlled at 25 ± 2 ℃, and humidity is controlled at 85% ± 2%, avoids direct sunlight, until scion and stock heal fully, grafting survives.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103749158A (en) * | 2014-01-07 | 2014-04-30 | 四川胜泽源农业投资有限公司 | Micro-grafting method for acer palmatum and acer paimatum under tissue culture condition |
| CN104025911A (en) * | 2014-05-15 | 2014-09-10 | 湖北圭萃园农林股份有限公司 | Method for cultivating walnut seedling with container in greenhouse |
| CN104137739A (en) * | 2013-05-10 | 2014-11-12 | 孔令波 | Thin-shell walnut double-tongue grafting technology |
| CN104304007A (en) * | 2014-10-08 | 2015-01-28 | 广东药学院 | In-vitro culture method for berchemia lineata |
| CN104904499A (en) * | 2014-05-09 | 2015-09-16 | 常州市金土地农牧科技服务有限公司 | Method for grafting caryaillinoinensis |
| CN104957040A (en) * | 2015-07-16 | 2015-10-07 | 句容市容北茶文化有限公司 | Carya illinoensis cell embryo tissue culture method |
| CN108966891A (en) * | 2018-09-30 | 2018-12-11 | 四川省农业科学院园艺研究所 | A method of preparing Lee, the sterile scion of apricot silver leaf |
| CN109496773A (en) * | 2018-12-26 | 2019-03-22 | 云南省林业科学院漾濞核桃研究院 | Apocarya Light media small container grafting Raining season wooding method |
| CN110192528A (en) * | 2019-07-01 | 2019-09-03 | 南京林业大学 | A kind of blue or green money willow adventitious bud inducing and shoot proliferation cultural method |
| CN110810251A (en) * | 2019-12-24 | 2020-02-21 | 南京林业大学 | Micro-grafting method for apocarya |
| CN114793659A (en) * | 2022-05-19 | 2022-07-29 | 北京林业大学 | Improved variety rapid propagation method for mature chestnut plants |
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| CN104137739A (en) * | 2013-05-10 | 2014-11-12 | 孔令波 | Thin-shell walnut double-tongue grafting technology |
| CN103749158A (en) * | 2014-01-07 | 2014-04-30 | 四川胜泽源农业投资有限公司 | Micro-grafting method for acer palmatum and acer paimatum under tissue culture condition |
| CN103749158B (en) * | 2014-01-07 | 2015-07-01 | 四川胜泽源农业投资有限公司 | Micro-grafting method for acer palmatum and acer paimatum under tissue culture condition |
| CN104904499A (en) * | 2014-05-09 | 2015-09-16 | 常州市金土地农牧科技服务有限公司 | Method for grafting caryaillinoinensis |
| CN104025911A (en) * | 2014-05-15 | 2014-09-10 | 湖北圭萃园农林股份有限公司 | Method for cultivating walnut seedling with container in greenhouse |
| CN104304007A (en) * | 2014-10-08 | 2015-01-28 | 广东药学院 | In-vitro culture method for berchemia lineata |
| CN104957040A (en) * | 2015-07-16 | 2015-10-07 | 句容市容北茶文化有限公司 | Carya illinoensis cell embryo tissue culture method |
| CN108966891A (en) * | 2018-09-30 | 2018-12-11 | 四川省农业科学院园艺研究所 | A method of preparing Lee, the sterile scion of apricot silver leaf |
| CN108966891B (en) * | 2018-09-30 | 2020-11-10 | 四川省农业科学院园艺研究所 | Method for preparing aseptic scions of plum and apricot |
| CN109496773A (en) * | 2018-12-26 | 2019-03-22 | 云南省林业科学院漾濞核桃研究院 | Apocarya Light media small container grafting Raining season wooding method |
| CN110192528A (en) * | 2019-07-01 | 2019-09-03 | 南京林业大学 | A kind of blue or green money willow adventitious bud inducing and shoot proliferation cultural method |
| CN110192528B (en) * | 2019-07-01 | 2020-10-16 | 南京林业大学 | Method for inducing adventitious buds of cyclocarya paliurus and subculture multiplication culture |
| CN110810251A (en) * | 2019-12-24 | 2020-02-21 | 南京林业大学 | Micro-grafting method for apocarya |
| CN114793659A (en) * | 2022-05-19 | 2022-07-29 | 北京林业大学 | Improved variety rapid propagation method for mature chestnut plants |
| CN114793659B (en) * | 2022-05-19 | 2023-11-03 | 北京林业大学 | Improved variety rapid propagation method for adult chestnut plants |
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Application publication date: 20130403 |