CN101138319B - Tristellateia australasiae tissue culturing and propagating method - Google Patents
Tristellateia australasiae tissue culturing and propagating method Download PDFInfo
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- CN101138319B CN101138319B CN2007100305940A CN200710030594A CN101138319B CN 101138319 B CN101138319 B CN 101138319B CN 2007100305940 A CN2007100305940 A CN 2007100305940A CN 200710030594 A CN200710030594 A CN 200710030594A CN 101138319 B CN101138319 B CN 101138319B
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Abstract
The present invention relates to a tissue culture and propagating method of Tristellateia australasiae A.Richard; the young leaves bourgeoning at the base of robust Tristellateia australasiae A.Richard are selected as the explants in the growing season; and after being sterilized, the explants are inoculated into an MS culture medium with 0.1 to 0.5 mg 6-benzyl purine and 0.01 to 0.05 mg seradix every liter until the indefinite but is sprouted; after being induced and cut, the indefinite bud is cultivated in the same culture medium for propagation in the pattern of successive transfer culture;when the bud height is 2 - 3 cm, the bud is cut to inoculated into the MS radication culture medium with 0.5 - 1.0 mg of seradix and 0.05 - 1 mg of active carbon every liter until the bud is normallyredicated; at last, when the tube seedling grows to 3 to 5 cm, the seedling is trained in the natural light and then transplanted. The method of the present invention can propagate the tube seedlingsinmass with a radication rate of 100 percent and a survival rate of more than 90 percent; in addition, the seedlings are prevented from disease and insect pest and can grow healthily; the quality isexc ellent; the resistance is strong and the growth is fast.
Description
Technical field
The present invention relates to the process for breeding and culturing of star fruit rattan, is to utilize tissue culture technique to breed the method for star fruit rattan specifically.
Background technology
Star fruit rattan is the evergreen climing property shrub of Malpighiaceae Tristellateia, is distributed in China Taiwan Province, Philippine, Malaysia, Austronesia etc.The star fruit rattan plant ability of overgrowing is strong, and riotous growth, leaf are to life, and the close life of flower is yellow flower, the florescence 4-5 month (or annual), the fruit phase 5-7 month (or annual).Look brown when samara was ripe, and the name of star fruit rattan comes from its samara as the star shape.This kind is rare woody climber, and drought-enduring, wind resistance, happiness sunlight are the lianes that desirable rattan teapoy is planted.Cuttage is used in the routine breeding of star fruit rattan, but reproduction speed is slower, is difficult to obtain a large amount of seedlings in a short time and satisfies market demand.Directly to buy star fruit rattan seedling price higher from external, thereby press for the application organizes cultural method star fruit rattan is carried out fast breeding technique and batch production production seedling.
Summary of the invention
The objective of the invention is to develop a kind of method that can breed star fruit rattan efficiently.
Our totipotency and plant tissue culture technique by plant tissue cell utilizes drawing materials, sterilize, cultivating of explant, and the rooting rate of the star fruit rattan of cultivating in this way reaches 100%, and survival rate is more than 90%, thereby realized purpose of the present invention.
The tissue culture propagation of star fruit rattan of the present invention, its feature comprises the steps:
(1) explant induction materials disinfection and adventitious bud inducing: choosing the young tender budlet that star fruit rattan (Tristellateia australasiae A.Richard) the plant base portion of robust growth sprouts out in the season of growth is explant, water is rinsed the back well earlier with alcohol-pickled 10～30s of 70%～75%, again with the liquor natrii hypochloritis of 0.1%～0.2% mercuric chloride solution or 1%～2% 10～15min that sterilizes, downcut the stem section of the terminal bud or the band joint of band growing point behind the aseptic water washing, be inoculated in the medium, in temperature (28 ± 2) ℃, illuminance 1500～2000lx, illumination 8～12h/ days, cultivate and form indefinite bud, described medium contains 6-benzyl purine 0.1～0.5mg for every liter, indolebutyric acid 0.01～0.05mg, sucrose 25～30g and agar 5～7g, all the other are MS, pH 5.8～6.0;
(2) shoot proliferation of indefinite bud: successive transfer culture is in the medium identical with step (1) again after the indefinite bud cutting that step (1) is induced, and in temperature (28 ± 2) ℃, illuminance 1500～2000lx carried out the propagation of indefinite bud in illumination 8～12h/ days;
(3) culture of rootage: when the high about 2～3cm of step (2) bud, cutting-out is inoculated in the root media, in temperature (28 ± 2) ℃, illuminance 1500～2000lx was cultured to normally and takes root in illumination 8～12h/ days, contained indolebutyric acid 0.5～1.0mg in every liter of the described medium, active carbon 0.05～0.1g, sucrose 25～30g and agar 5～7g, all the other are MS, pH 5.8～6.0;
(4) test-tube seedling transplanting: when the rooting tube plantlet of step (3) grows to 3～5cm, natural lighting lower refining seedling 7～10 days, clean the root medium, be transplanted in the matrix that vermiculite and peat soil and perlite mixed in equal amounts form, spraying and moisturizing 85%～95%, shade 75%～85%, 20～30 ℃ of cultivations of insulation.
The described aseptic water washing number of times of step (1) is 4～5 times, the stem segment length of the terminal bud of described band growing point or band joint is about 1cm, for alleviating brownization, cultivate after 25～30 days earlier and change over to again in the same fresh culture, cut a new otch at the culture materials base portion, continue to cultivate 25～35 days energy and on explant, form indefinite bud.
A shoot proliferation cycle in the step (2) is 30～35 days, and the propagation multiple of all after dates of each shoot proliferation is 3～4 times, and subculture still can keep the rate of increase after 10 generations.
The described incubation time of step (3) generally is 10～15 days, rooting rate can reach 100% in the time of 15 days, and every young plant has 3～5 of radicals, normally elongation growth of root when not adding active carbon in the root media, but the activated carbon excessive concentration then can suppress the generation of adventive root, reduces rooting rate.
Transplant to be generally after 40～50 days in the step (4) and can go up potted plant training.
MS in the above-mentioned medium is international medium, its composition and compound method referring to document (Tan Wencheng, Dai Cegang chief editor. the ornamental plants tissue culture technique. Beijing: China Forest publishing house, 1991.).
The present invention adopts plant tissue culture technique can breed seedling fast, obtain a large amount of test-tube plantlets, the rooting rate of star fruit rattan can reach 100%, survival rate can reach more than 90%, and can be regularly, decide matter, quantitatively provide the test-tube plantlet of the standardized neat and consistent of specification to meet the need of market, worm robust growth that test-tube plantlet is anosis, quality better, resistance, growth are rapid, be convenient to production management and reduce production costs, for the needs that satisfy seedling market provide a valid approach.
Embodiment
Following embodiment further specifies of the present invention, is not limitation of the present invention.
Embodiment 1:
Choosing the long young tender budlet of 3～5cm that star fruit rattan (Tristellateia australasiae A.Richard) the plant base portion of robust growth sprouts out in the season of growth is explant, after under running water, rinsing well, earlier in 70% alcohol, soak 30s, again with 0.2% mercuric chloride solution sterilization 10 minutes, aseptic water washing 4 times, be cut into terminal bud that is about 1cm band growing point or the stem section of being with joint, (every liter contains 6-benzyl purine (6-BA) 0.1mg to be inoculated into medium, indolebutyric acid 0.01mg, sucrose 25g and agar 5g, all the other are MS, pH 5.8), 28 ℃ of temperature, illuminance 1500lx illumination 12h/ days, changes in the new same above-mentioned medium again after 25 days, cut a new otch at the culture materials base portion, cultivate 25 days energy again and on explant, form indefinite bud.With after the cutting of the indefinite bud that induces again successive transfer culture in medium same as described above, 28 ℃ of temperature, illuminance 1500lx, carried out the propagation of indefinite bud in illumination 12h/ days, 30 days is 1 shoot proliferation cycle, the propagation multiple of an all after date of shoot proliferation is 3 times, and repeatedly subculture is bred multiple behind the continuation subculture and slightly raise.When the high about 2～3cm of bud, cutting-out is inoculated into root media, and (every liter contains indolebutyric acid 0.5mg, active carbon 0.1g, sucrose 25g and agar 5g, all the other are MS, pH 5.8), at 28 ℃ of temperature, illuminance 1500l x, illumination 12h/ days, make normally and take root, rooting rate 100% when cultivating 15 days, every young plant has 3～5 of radicals.When test-tube plantlet grows to 3～5cm, natural lighting lower refining seedling 7 days, open bottle stopper then, with tweezers test-tube plantlet is taken out from blake bottle, clean the root medium, be transplanted in the matrix that forms by vermiculite, peat soil and perlite mixed in equal amounts, spraying and moisturizing 85%, shade 85%, at 28 ℃, survival rate 90%.Transplant and to go up potted plant training in back 40 days.
Embodiment 2:
Choosing the long young tender budlet of 3～5cm that star fruit rattan (Tristellateia australasiae A.Richard) the plant base portion of robust growth sprouts out in the season of growth is explant, after under running water, rinsing well, earlier in 75% alcohol, soak 10s, again with 0.1% mercuric chloride solution sterilization 15min, aseptic water washing 5 times, be cut into terminal bud that is about 1cm band growing point or the stem section of being with joint, (every liter contains 6-benzyl purine 0.5mg to be inoculated into medium, indolebutyric acid 0.05mg, sucrose 30g and agar 7g, all the other are MS, pH 6.0), 26 ℃ of temperature, illuminance 2000lx illumination 8h/ days, cultivates and changes in the same fresh culture after 30 days again, cut a new otch at the culture materials base portion, cultivate 30 days energy again and on explant, form indefinite bud.With after the cutting of the indefinite bud that induces again successive transfer culture in medium same as described above, at 26 ℃ of temperature, illuminance 2000lx, illumination 8h/ days, carry out the propagation of indefinite bud, 35 days is 1 shoot proliferation cycle, the propagation multiple of an all after date of shoot proliferation is 4 times, repeatedly subculture.When the high about 2～3cm of bud, cutting-out is inoculated into root media, and (every liter contains indolebutyric acid 1.0mg, active carbon 0.05g, sucrose 30g and agar 7g, all the other are MS, pH 6.0), at 26 ℃ of temperature, illuminance 2000lx, illumination 8h/ days, make normally and take root, rooting rate can reach 90% when cultivating 10 days, and every young plant has 3～5 of radicals.When test-tube plantlet grows to 3～5cm, natural lighting lower refining seedling 10 days, open bottle stopper then, with tweezers test-tube plantlet is taken out from blake bottle, clean the root medium, be transplanted in the matrix that forms by vermiculite, peat soil and perlite mixed in equal amounts, spraying and moisturizing 90%, shade 75%, at 25 ℃, survival rate 95%.Transplant and to go up potted plant training in back 50 days.
Embodiment 3:
Choosing the long young tender budlet of 3～5cm that star fruit rattan (Tristellateia australasiae A.Richard) the plant base portion of robust growth sprouts out in the season of growth is explant, after under running water, rinsing well, earlier in 70% alcohol, soak 30s, use 1.5% liquor natrii hypochloritis's solution disinfection 10min again, aseptic water washing 5 times, be cut into terminal bud that is about 1cm band growing point or the stem section of being with joint, (every liter contains 6-benzyl purine 0.3mg to be inoculated into medium, indolebutyric acid 0.03mg, sucrose 30g and agar 5g, all the other are MS, pH 6.0), 30 ℃ of temperature, illuminance 1800lx illumination 10h/ days, changes over to after 25 days in the new same above-mentioned medium again, cut a new otch at the culture materials base portion, cultivate 35 days energy again and on explant, form indefinite bud.With after the cutting of the indefinite bud that induces again successive transfer culture in medium same as described above, at 30 ℃ of temperature, illuminance 1800lx, illumination 10h/ days, carry out the propagation of indefinite bud, 30 days is 1 shoot proliferation cycle, the propagation multiple of an all after date of shoot proliferation is 3 times, can carry out repeatedly subculture.When the high about 2～3cm of bud, cutting-out is inoculated into root media, and (every liter contains indolebutyric acid 0.7mg, active carbon 0.05g, sucrose 30g and agar 5g, all the other are MS, pH 6.0), at 30 ℃ of temperature, illuminance 1800lx, illumination 10h/ days, make normally and take root, rooting rate can reach 95% when cultivating 13 days, and every young plant has 3～5 of radicals.When test-tube plantlet grows to 2～3cm, natural lighting lower refining seedling 10 days, open bottle stopper then, with tweezers test-tube plantlet is taken out from blake bottle, clean the root medium, be transplanted in the matrix that forms by vermiculite, peat soil and perlite mixed in equal amounts, spraying and moisturizing 95%, shade 80%, at 28 ℃, survival rate 90%.Transplant and to go up potted plant training in back 45 days.
Claims (2)
1. the tissue culture propagation of star fruit rattan, its feature comprises the steps:
(1) explant induction materials disinfection and adventitious bud inducing: choosing the young tender budlet that star fruit rattan (Tristellateia australasiae A.Richard) the plant base portion of robust growth sprouts out in the season of growth is explant, water is rinsed the back well earlier with alcohol-pickled 10～30s of 70%～75%, again with 0.1%～0.2% mercuric chloride solution sterilization, 10～15min or with 1.5% the liquor natrii hypochloritis 15min that sterilizes, downcut the terminal bud of band growing point behind the aseptic water washing, be inoculated in the medium, in temperature (28 ± 2) ℃, illuminance 1500～2000lx, illumination 8～12h/ days, cultivate and form indefinite bud, described medium contains 6-benzyl purine 0.1～0.5mg for every liter, indolebutyric acid 0.01～0.05mg, sucrose 25～30g and agar 5～7g, all the other are MS, pH5.8～6.0;
(2) shoot proliferation of indefinite bud: successive transfer culture is in the medium identical with step (1) again after the indefinite bud cutting that step (1) is induced, and in temperature (28 ± 2) ℃, illuminance 1500～2000lx carried out the propagation of indefinite bud in illumination 8～12h/ days;
(3) culture of rootage: when the high about 2～3cm of step (2) bud, cutting-out is inoculated in the root media, in temperature (28 ± 2) ℃, illuminance 1500～2000lx, be cultured to normally in illumination 8～12h/ days and take root, contain indolebutyric acid 0.5～1.0mg and active carbon 0.05～0.1g, sucrose 25～30g and agar 5～7g in every liter of the described medium, all the other are MS, pH5.8～6.0;
(4) test-tube seedling transplanting: when the rooting tube plantlet of step (3) grows to 3～5cm, natural lighting lower refining seedling 7～10 days, clean the root medium, be transplanted in the matrix that vermiculite and peat soil and perlite mixed in equal amounts form, spraying and moisturizing 85%～95%, shade 75%～85%, 20～30 ℃ of cultivations of insulation.
2. according to the tissue culture propagation of the star of claim 1 fruit rattan, it is characterized in that the described aseptic water washing number of times of step (1) is 4～5 times, the terminal bud cultivation earlier of band growing point is changed in the same fresh culture after 25～30 days again, cut a new otch at the culture materials base portion, continue to cultivate 25～35 days energy and on explant, form indefinite bud; A shoot proliferation cycle in the step (2) is 30～35 days; The described incubation time of step (3) is 10～15 days, transplants in the step (4) and goes up potted plant training after 40～50 days.
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| CN102187810B (en) * | 2010-03-16 | 2012-10-10 | 中国科学院华南植物园 | Tissue culture propagation method for curcuma soloensis |
| CN114586685A (en) * | 2022-03-21 | 2022-06-07 | 玉林师范学院 | Culture method of Chinese starjasmine stem virus-free seedlings |
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| WO2002063023A2 (en) * | 2001-02-05 | 2002-08-15 | The University Of South Carolina Research Foundation | Sustained totipotent regenerable tissue culture of arundo donax (giant reed) and totipotent tissue and plants produced therefrom |
| WO2003006672A2 (en) * | 2001-07-10 | 2003-01-23 | D-Squared Biotechnologies, Inc. | Antimicrobial nucleic acid antibodies, and materials and methods for making and using same |
| CN1166286C (en) * | 2003-04-14 | 2004-09-15 | 中国科学院华南植物研究所 | Tissue culture reproduction method of snake grass |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2002063023A2 (en) * | 2001-02-05 | 2002-08-15 | The University Of South Carolina Research Foundation | Sustained totipotent regenerable tissue culture of arundo donax (giant reed) and totipotent tissue and plants produced therefrom |
| WO2003006672A2 (en) * | 2001-07-10 | 2003-01-23 | D-Squared Biotechnologies, Inc. | Antimicrobial nucleic acid antibodies, and materials and methods for making and using same |
| CN1166286C (en) * | 2003-04-14 | 2004-09-15 | 中国科学院华南植物研究所 | Tissue culture reproduction method of snake grass |
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