CN101536674A - Tissue culture reproduction method of Chinese evergreen - Google Patents
Tissue culture reproduction method of Chinese evergreen Download PDFInfo
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- CN101536674A CN101536674A CN200910038981A CN200910038981A CN101536674A CN 101536674 A CN101536674 A CN 101536674A CN 200910038981 A CN200910038981 A CN 200910038981A CN 200910038981 A CN200910038981 A CN 200910038981A CN 101536674 A CN101536674 A CN 101536674A
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Abstract
The invention discloses a tissue culture reproduction method of Chinese evergreen and aims to provide a tissue culture reproduction method of the Chinese evergreen with high reproduction speed, mass test seedling obtainment in a short term, little damage to a mother plant and the maintenance of mother plant fine traits. The method comprises the following steps: selecting young stems of eugenic Chinese evergreen as explants, disinfecting the explants, carrying out the adventitious-bud induction of an induction culture medium and coupling adventitious buds into a reproduction culture medium for successive transfer reproduction; cutting and inoculating the buds to a rooting culture medium for rooting culture while the buds grow to the height of 3cm to 5cm; training the seedlings under natural light for 7 days to 10 days when the test seedlings grow to the height of 6cm to 7cm; and taking the test seedlings out, washing the rooting culture medium, planting in a volumetric mixture medium comprising sand, perlite, peat soil, and the like and obtaining planting seedlings after conventional culture. The invention can provide markets with mass Chinese evergreen test seedlings.
Description
Technical field:
The present invention relates to the tissue culture propagation of thick rib grass.
Background technology:
Thick rib grass is the common name that the thick rib grass of Araeceae belongs to (Aglaonema) plant, originates in the ground such as India, Thailand, Vietnam, Philippine, Malaysia and Indonesia of the southeast, Asia, is the perennial evergreen herbaceous plant.20～150 centimetres of thick rib grass plant heights.The leaf alternate, lanceolar is extremely narrow avette, long 10～45 centimetres of leaf, leaf is wide 4～16 centimetres.Spend little not obviously, inflorescence is a spadix, spathe white or green white, and fruit is a berry, can become redness when ripe.Thick rib grass ornamental value height is a kind of good watch leaf flower, can be used as high-grade ornamental foliage plant and is used for indoor and the flower garden is viewed and admired.
Three kinds of modess of reproduction such as the available plant division of thick rib grass, cuttage and seminal propagation, division propagation mode speed is slow, though and seminal propagation is the effective means of seed selection new varieties, but required time is long, to becoming the strain time to need the time in 2～3 years, and offspring Yi Xingzhuan separates by seed germination, and conventional breeding is main modes of reproduction with terminal bud and two kinds of cuttages of stem section mostly in producing at present, but this reproduction speed is also limited, and very big to the good maternal plant damage of proterties.
Do not see at present and thick rib grass is carried out tissue culture, the report of seedling large-scale production and patent application.
Summary of the invention:
It is fast to the invention provides a kind of reproduction speed, can obtain a large amount of test-tube plantlets in a short time, and little to the maternal plant damage, can keep the tissue culture propagation of the thick rib grass of maternal plant merit.
Histiocytic totipotency of the main appliable plant of the present invention and plant tissue culture technique are selected suitable explant, and sterilization method is realized the object of the invention thereby unique medium carries out the tissue culture of thick rib grass.
The tissue culture propagation of thick rib grass of the present invention may further comprise the steps:
A. material is selected: choosing the young stem of eugonic thick rib grass is explant;
B. inducing of the sterilization of explant and indefinite bud: explant is washed, be to soak in 70%～80% alcohol 30～60 seconds in volume fraction earlier, be 0.1%～0.2% mercuric chloride solution sterilization 8～10 minutes with the quality concentration expressed in percentage by volume again, aseptic water washing 4～5 times, be cut into sections, be inoculated in the inducing culture, take out after 5～7 days, be 0.1%～0.2% mercuric chloride solution sterilization 8～10 minutes with the quality concentration expressed in percentage by volume once more, aseptic water washing 4～5 times, change in the new inducing culture, on explant, form indefinite bud, during being every liter, described inducing culture contains 1.0～5.0 milligrams of 6-benzyl purines, 0.2～1.0 milligram of methyl, sucrose 20～30 grams, agar 6～7 grams, all the other compositions are MS;
C. shoot proliferation: the propagation of the indefinite bud cutting back successive transfer culture that induces being carried out indefinite bud in proliferated culture medium, contain 5.0～10.0 milligrams of 6-benzyl purines, 0.2～1.0 milligram of methyl, sucrose 20～30 grams, agar 6～7 grams during described proliferated culture medium is every liter, all the other compositions are MS;
D. culture of rootage: when high 3～5 centimetres high of bud, cutting-out is inoculated on the root media, described root media is every liter and contains 0.2～2.0 milligram of 6-benzyl purine, 0.5～1.0 milligram of methyl, active carbon 0.5～1 gram, sucrose 20～30 grams, agar 6～7 grams that all the other compositions are 1/2MS;
E. test-tube plantlet is transplanted: when test-tube plantlet grows to 6～7 centimetres high, natural lighting lower refining seedling 7～10 days, take out test-tube plantlet, wash the root medium off, plant by in sand, perlite, the composite matrix of peat soil equal-volume, obtain cultivating seedling after the routine cultivation;
In above-mentioned a, b, four steps of c, d, used medium (inducing culture, proliferated culture medium and root media) pH 5.5～5.8; Condition of culture: 24～30 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day.
Test-tube plantlet after transplanting among the present invention is noted watering, shades, is incubated, and survival rate can reach more than 85%, just can go up potted plant training after general 40 days.
Usefulness aseptic water washing of the present invention 4～5 times mainly is under aseptic environments, and mercuric chloride is rinsed well.
The sections that is cut into described in the present invention is that the explant that aseptic water washing is clean preferably is cut into the sections about 1 centimetre with after the mercuric chloride sterilization.
The present invention is applicable to the tissue culture propagating of thick rib grass platymiscium.
Described MS is international medium, its composition and compound method are edited referring to Tan Wencheng, Dai Cegang, " ornamental plants tissue culture technique ", Beijing: China Forest publishing house, 1991, described 1/2MS reduces by half macroelement among the MS and trace element, other components unchanged and the medium that forms.
Of the present invention at the natural lighting lower refining seedling, its condition is survival and the growth not influence seedling as conditions such as intensity of illumination, temp. and humidity.
Use explant sterilization method of the present invention, the sterilization success rate can reach 70～80%, 50～60 days energy and form indefinite bud on explant.Can rise in value to indefinite bud by subculture increment method of the present invention, general 45 days is 1 shoot proliferation cycle, and the propagation multiple of the indefinite bud of all after dates of per 1 shoot proliferation is 2～4 times.Indefinite bud can normally be taken root in root media of the present invention, and rooting rate can reach more than 90% in the time of 30 days, and every young plant has 3～5 of radicals.
Beneficial effect of the present invention is as follows:
The present invention only needs simple Plant Tissue Breeding equipment just can finish in 120 days from the one-period of the formation of choosing test-tube plantlet of explant, accelerated the reproduction speed of thick rib grass greatly, rise in value by subculture, each subculture cycle can rise in value 2～4 times to indefinite bud, therefore can obtain a large amount of test-tube plantlets at short notice, it is high that the test-tube plantlet that application the present invention turns out has stability, Miao Zhuan, the seedling quality better, resistance, well-grown characteristics, survival rate can reach more than 85%, and it is fast to emerge, and can satisfy the demand in market in a large number, and this method is little to the maternal plant damage, can keep the merit of maternal plant.
Embodiment:
Embodiment 1
A. material is selected: choosing the thick rib grass of roundleaf (Aglaonema rotundum) tender stem segments in the season of growth is explant.
B. inducing of the sterilization of explant and indefinite bud: after the tender stem segments of the thick rib grass of roundleaf is rinsed well under running water, be to soak 30 seconds in 70% alcohol in volume fraction earlier, be 0.1% mercuric chloride solution sterilization 8 minutes with the quality concentration expressed in percentage by volume again, aseptic water washing 4 times, be cut into sections long about 1 centimetre, be inoculated on the inducing culture, take out sections after 5 days once more with quality concentration expressed in percentage by volume 0.1% mercuric chloride solution sterilization 8 minutes, aseptic water washing 5 times, after change in the new inducing culture, the sterilization success rate is 71%, energy formed indefinite bud on explant in 60 days, during being every liter, described inducing culture contains 2.0 milligrams of 6-benzyl purines, 0.2 milligram of methyl, sucrose 20 grams, agar 6 grams, all the other compositions are MS;
C. shoot proliferation: with after the indefinite bud cutting that induces again successive transfer culture on proliferated culture medium, carry out the propagation of indefinite bud, general 45 days is 1 shoot proliferation cycle, the propagation multiple of all after dates of per 1 shoot proliferation is 2.5 times, contain 5.0 milligrams of promising 6-benzyl purines, 0.5 milligram of methyl, sucrose 20 grams, agar 7 grams in every liter of the described proliferated culture medium, all the other compositions are MS;
D. culture of rootage: when high about 3～5 centimetres high of bud, cutting-out is inoculated on the root media, indefinite bud can normally be taken root, rooting rate can reach 90%, every young plant has 4.5 of radicals, described root media is every liter and contains 1.0 milligrams of 6-benzyl purines, 0.5 milligram of methyl, active carbon 0.5 gram, sucrose 20 grams, agar 7 grams that all the other compositions are 1/2MS
E. test-tube seedling transplanting: when test-tube plantlet grows to 6～7 centimetres high, natural lighting lower refining seedling 9 days.Open bottle stopper then, test-tube plantlet is taken out from blake bottle, wash the root medium off, plant by in sand, perlite, the composite matrix of peat soil equal-volume with tweezers.
Used medium pH 5.5-5.8 in above-mentioned a, b, c, d step; Condition of culture: 24～30 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day.
Transplanted seedling is noted watering, shades, is incubated, and survival rate can reach more than 85%, and transplant the back and can go up potted plant training after about 40 days,
From the general time that needs 120 days of the acquisition of choosing test-tube plantlet of explant.
Embodiment 2
A. material is selected: choosing the thick rib grass of silver-colored king (Aglaonema Silver King) tender stem segments in the season of growth is explant.
B. inducing of the sterilization of explant and indefinite bud: after the tender stem segments of the thick rib grass of silver-colored king is rinsed well under running water, in 75% alcohol, soaked 45 seconds earlier, again with 0.15% mercuric chloride solution sterilization 10 minutes, aseptic water washing 5 times, be cut into sections long about 1 centimetre, be inoculated on the inducing culture, take out sections after 6 days, once more with 0.15% mercuric chloride solution sterilization 8 minutes, aseptic water washing 5 times, after change in the new inducing culture, the sterilization success rate can reach 78%, energy formed indefinite bud on explant in 60 days, contained 3.0 milligrams of 6-benzyl purines during described inducing culture is every liter, 0.5 milligram of methyl, sucrose 25 grams, agar 7 grams, all the other compositions are MS;
C. shoot proliferation: with after the indefinite bud cutting that induces again successive transfer culture on proliferated culture medium, carry out the propagation of indefinite bud, general 45 days is 1 shoot proliferation cycle, the propagation multiple of all after dates of per 1 shoot proliferation is 3 times, contain 7.0 milligrams of 6-benzyl purines, 0.5 milligram of methyl, sucrose 25 grams, agar 7 grams during described proliferated culture medium is every liter, all the other compositions are MS;
D. culture of rootage: when high about 3～5 centimetres high of bud, cutting-out is inoculated on the root media, indefinite bud can normally be taken root, rooting rate can reach 90%, every young plant has 5 of radicals, described root media is every liter and contains 0.5 milligram of 6-benzyl purine, 0.8 milligram of methyl, active carbon 0.75 gram, sucrose 25 grams, agar 6 grams that all the other compositions are 1/2MS
E. test-tube seedling transplanting: when test-tube plantlet grows to 6～7 centimetres high, natural lighting lower refining seedling 7 days.Open bottle stopper then, test-tube plantlet is taken out from blake bottle, wash the root medium off, plant by in sand, perlite, the composite matrix of peat soil equal-volume with tweezers.
Used medium pH 5.5～5.8 in above-mentioned a, b, c, d step, condition of culture: 24～30 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day.
Transplanted seedling is noted watering, shades, is incubated, and survival rate can reach more than 90%, transplants the back and can go up potted plant training after about 40 days.
From the general time that needs 115 days of the acquisition of choosing test-tube plantlet of explant.
Embodiment 3
A. material is selected: choosing the thick rib grass of exceedingly popular leaf (Aglaonema Tembaga Super Red) tender stem segments in the season of growth is explant.
B. inducing of the sterilization of explant and indefinite bud: after the tender stem segments of the thick rib grass of exceedingly popular leaf is rinsed well under running water, in 80% alcohol, soaked 60 seconds earlier, again with 0.2% mercuric chloride solution sterilization 8 minutes, aseptic water washing 5 times, be cut into sections long about 1 centimetre, be inoculated on the inducing culture, take out sections after 7 days, once more with 0.1% mercuric chloride solution sterilization 8 minutes, aseptic water washing 5 times, after change in the new inducing culture, the sterilization success rate can reach 79%, energy formed indefinite bud on explant in 50 days, contained 5.0 milligrams of 6-benzyl purines during described inducing culture is every liter, 1.0 milligrams of methyls, sucrose 30 grams, agar 7 grams, all the other compositions are MS;
C. shoot proliferation: with after the indefinite bud cutting that induces again successive transfer culture on proliferated culture medium, carry out the propagation of indefinite bud, general 45 days is 1 shoot proliferation cycle, the propagation multiple of all after dates of per 1 shoot proliferation is 4 times, contain 10.0 milligrams of promising 6-benzyl purines, 1.0 milligrams of methyls, sucrose 30 grams, agar 6 grams in every liter of the described proliferated culture medium, all the other compositions are MS;
D. culture of rootage: when high about 3～5 centimetres high of bud, cutting-out is inoculated on the root media, indefinite bud can normally be taken root, rooting rate can reach more than 95%, every young plant has 7 of radicals, described root media is every liter and contains 2.0 milligrams of 6-benzyl purines, 1.0 milligrams of methyls, active carbon 1 gram, sucrose 30 grams, agar 7 grams that all the other compositions are 1/2MS;
E. test-tube seedling transplanting: when test-tube plantlet grows to 6～7 centimetres high, natural lighting lower refining seedling 10 days.Open bottle stopper then, test-tube plantlet is taken out from blake bottle, wash the root medium off, plant by in sand, perlite, the composite matrix of peat soil equal-volume with tweezers.
Used medium pH 5.5～5.8 in above-mentioned a, b, c, d step, condition of culture: 24～30 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12 hours/day.
Transplanted seedling is noted watering, shades, is incubated, and survival rate can reach more than 95%, transplants the back and can go up potted plant training after about 40 days.
From the general time that needs 110 days of the acquisition of choosing test-tube plantlet of explant.
Claims (3)
1. the tissue culture propagation of a thick rib grass is characterized in that may further comprise the steps:
A. material is selected: choosing the young stem of eugonic thick rib grass is explant;
B. inducing of the sterilization of explant and indefinite bud: explant is washed, be to soak in 70%～80% alcohol 30～60 seconds in volume fraction earlier, be 0.1%～0.2% mercuric chloride solution sterilization 8～10 minutes with the quality concentration expressed in percentage by volume again, aseptic water washing 4～5 times, be cut into sections, be inoculated in the inducing culture, take out after 5～7 days, be 0.1%～0.2% mercuric chloride solution sterilization 8～10 minutes with the quality concentration expressed in percentage by volume once more, aseptic water washing 4～5 times, change in the new inducing culture, on explant, form indefinite bud, during being every liter, described inducing culture contains 1.0～5.0 milligrams of 6-benzyl purines, 0.2～1.0 milligram of methyl, sucrose 20～30 grams, agar 6～7 grams, all the other compositions are MS;
C. shoot proliferation: the indefinite bud cutting back successive transfer culture that induces is carried out the increment of indefinite bud in proliferated culture medium, contain 5.0～10.0 milligrams of 6-benzyl purines, 0.2～1.0 milligram of methyl, sucrose 20～30 grams, agar 6～7 grams during described proliferated culture medium is every liter, all the other compositions are MS;
D. culture of rootage: when high 3～5 centimetres high of bud, cutting-out is inoculated on the root media, described root media is every liter and contains 0.2～2.0 milligram of 6-benzyl purine, 0.5～1.0 milligram of methyl, active carbon 0.5～1 gram, sucrose 20～30 grams, agar 6～7 grams that all the other compositions are 1/2MS;
E. test-tube plantlet is transplanted: when test-tube plantlet grows to 6～7 centimetres high, natural lighting lower refining seedling 7～10 days, take out test-tube plantlet, wash the root medium off, plant by in sand, perlite, the composite matrix of peat soil equal-volume, obtain cultivating seedling after the routine cultivation;
In above-mentioned a, b, c, d step, used medium pH 5.5～5.8; Condition of culture: 24～30 ℃ of cultivation temperature, intensity of illumination 1500～2000lx, illumination 12 hours/day.
2. the tissue culture propagation of thick rib grass according to claim 1 is characterized in that the sections that is cut in the described b step is the sections that explant is cut into 1 centimeter length.
3. the tissue culture propagation of thick rib grass according to claim 1 and 2 is characterized in that the described explant of a step is the tender stem segments of the thick rib grass of roundleaf Aglaonema rotundum, the thick rib grass Aglaonema Silver King of silver-colored king or the thick rib grass of exceedingly popular leaf Aglaonema Tembaga Super Red.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102440193A (en) * | 2011-10-18 | 2012-05-09 | 东莞市生物技术研究所 | Method for inducing chill-proof Aglaonema spp polyploidy |
| CN103975853A (en) * | 2014-04-23 | 2014-08-13 | 地缘（厦门）生物科技有限公司 | Colorized aglaonema tissue culture method |
| CN106613065A (en) * | 2016-09-18 | 2017-05-10 | 大顺国际花卉股份有限公司 | Method for promoting tillering of Aglaonemaspp |
| CN107711513A (en) * | 2017-11-24 | 2018-02-23 | 广东省农业科学院环境园艺研究所 | A kind of thick rib grass quick breeding method for tissue culture |
| CN109220804A (en) * | 2018-11-05 | 2019-01-18 | 吴子平 | A kind of high efficiency quick breeding method of the thick rib grass of coloured silk leaf |
| CN109566418A (en) * | 2019-01-28 | 2019-04-05 | 三明耿道理生物科技有限公司 | A kind of thick strangle careless tissue culture propagation |
| CN110249953A (en) * | 2019-07-17 | 2019-09-20 | 景维杰 | The color thick rib grass of leaf is promoted to watch color appearance technology |
| CN111972287A (en) * | 2020-07-21 | 2020-11-24 | 广州百德园艺有限公司 | Tissue culture method for spanish burley grass |
| CN114667930A (en) * | 2022-04-12 | 2022-06-28 | 广州花卉研究中心 | Method for changing growth amount and leaf color of leaves of tissue culture seedlings of ribgrass bicolor by using culture medium |
| CN114885839A (en) * | 2022-05-11 | 2022-08-12 | 中国科学院华南植物园 | Method for rapidly propagating seedlings of stem of common andrographis |
-
2009
- 2009-04-24 CN CN2009100389818A patent/CN101536674B/en not_active Expired - Fee Related
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102440193A (en) * | 2011-10-18 | 2012-05-09 | 东莞市生物技术研究所 | Method for inducing chill-proof Aglaonema spp polyploidy |
| CN103975853A (en) * | 2014-04-23 | 2014-08-13 | 地缘（厦门）生物科技有限公司 | Colorized aglaonema tissue culture method |
| CN103975853B (en) * | 2014-04-23 | 2016-04-27 | 地缘（厦门）生物科技有限公司 | The lucky method for tissue culture of a kind of colored thick rib grass |
| CN106613065A (en) * | 2016-09-18 | 2017-05-10 | 大顺国际花卉股份有限公司 | Method for promoting tillering of Aglaonemaspp |
| CN107711513A (en) * | 2017-11-24 | 2018-02-23 | 广东省农业科学院环境园艺研究所 | A kind of thick rib grass quick breeding method for tissue culture |
| CN109220804A (en) * | 2018-11-05 | 2019-01-18 | 吴子平 | A kind of high efficiency quick breeding method of the thick rib grass of coloured silk leaf |
| CN109566418A (en) * | 2019-01-28 | 2019-04-05 | 三明耿道理生物科技有限公司 | A kind of thick strangle careless tissue culture propagation |
| CN110249953A (en) * | 2019-07-17 | 2019-09-20 | 景维杰 | The color thick rib grass of leaf is promoted to watch color appearance technology |
| CN111972287A (en) * | 2020-07-21 | 2020-11-24 | 广州百德园艺有限公司 | Tissue culture method for spanish burley grass |
| CN114667930A (en) * | 2022-04-12 | 2022-06-28 | 广州花卉研究中心 | Method for changing growth amount and leaf color of leaves of tissue culture seedlings of ribgrass bicolor by using culture medium |
| CN114667930B (en) * | 2022-04-12 | 2023-02-21 | 广州花卉研究中心 | Method for changing growth amount and leaf color of leaves of tissue culture seedlings of ribgrass bicolor by using culture medium |
| CN114885839A (en) * | 2022-05-11 | 2022-08-12 | 中国科学院华南植物园 | Method for rapidly propagating seedlings of stem of common andrographis |
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