CN1739341A - Aseptic seedling tissue culturing and test tube seedling hardening off and transplating technology for anthurium andraeanum - Google Patents
Aseptic seedling tissue culturing and test tube seedling hardening off and transplating technology for anthurium andraeanum Download PDFInfo
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Abstract
The tissue culturing technology for anthurium andraeanum includes selecting small amount of aseptic seedling stem sections from tender tissue induction as the explant, and inoculating to culture medium I, II and III to induce synchronously callus, side bud and adventitious root; cutting test seedling after growing to 2-3 cm high, chemical solution soaking, transplanting, transferring to hardening off chamber and transplanting in survival rate over 95 %. The present invention makes it possible to form great amount of plants and is suitable for industrial seedling culturing.
Description
1.
Invention field
The present invention relates to field of plant tissue culture technique, particularly name is called the aseptic seedling tissue culture technique and the seedling exercising replanting technique for test-tube plantlet of peace ancestral flowering plant.In cultivation, select aseptic explant for use, on the medium of two kinds of different advantages, alternately cultivate, produce callus, lateral bud, adventive root synchronously, in the shortest time, form whole plant; Select for use the high whole plant with medicament of 2-3cm to soak to handle then and transplant, improve survival rate.The present invention creates good condition for the peace ancestral spends factorial seedling growth, and the present invention also helps the tissue culture worker that other plant carries out the batch production breeding.
2. background technology
2.1 tissue of anthurium culture technique
An Zuhua (Anthurium andreamum) originates in the torrid areas in America, is Araeceae peace ancestral Pittosporum perennial herb flowers, and flower is dazzlingly beautiful, and flower shape uniqueness has another name called the red palm, Huo Hehua, lamp stand flower, fancy candles lit in the bridal chamber at wedding, candle flowers.Because of it spends shape uniqueness, pattern is gorgeous and have wax gloss, very graceful lovely, and become the extremely top grade flower of people's welcome; And but the potted flower anniversary blooms, the cut-flowers in bottle life-span is long, is a kind of cut-flower material and potted plant kind in very great demand in the international market.The red palm becomes " nova " of ranking among International Flower market over nearly 5 years.
An Zuhua breeds with plant division and cuttage mode usually, and it is more and more to adopt tissue culture technique to grow seedlings at present, but the explant that major part is selected for use is stem apex, blade, petiole: stem apex differentiation rate height, growth is fast, because branch is few, suitable position of drawing materials is very few, is difficult to satisfy the demand of explant; Blade, petiole evoked callus, differentiation bud time are longer.From Pierk in 1974 cultivate first the peace ancestral spend achieve success since, the peace ancestral is spent the research report of callus induction both at home and abroad, be to breed a bud one this technology path of a whole plant of taking root mostly according to explant one callus one callus; To forming whole plant, need 130-180 days from the explant induction callus; And above technology all is to adopt this approach of organ generation type, callus subculture overlong time, and resistance is poor, easily catches an illness, and transplants to be difficult for surviving.
The objective of the invention is at the problems referred to above, provide a kind of and obtain the quick propagating technology of a large amount of test-tube plantlets in the shortest time, this technology has good stability, easy and simple to handle, proliferative speed advantage faster.Method of the present invention is according to explant one callus, bud, this technology path of adventive root one whole plant, and the whole plant that produces callus, bud, adventive root from explant induction only needed about 50-62 days.
Purpose of the present invention can realize by following technology:
The selection of A, explant: selecting for use the peace ancestral to spend 2～3cm height that young tender organ obtains through tissue culture and robust growth, growing way is even, quality is good aseptic seedling is material, and the stem section that cuts with 1～2 joint is that explant is cultivated.
The inoculation of B, explant: will be with the stem section of 1～2 joint, be seeded in I.1/2MS+6-BA0.5～1.0mg/L+NAA0.05mg/L+3% glucose+0.75% agar, II.1/2MS+6-BA1.5～2.0mg/L+NAA0.05mg/L+3% glucose+0.75% agar, III.1/2MS+6-BA1.0～1.5mg/L+NAA0.05mg/L+IAA0.05mg/L+3% glucose+0.75% agar, 10～12 stem sections of every bottle of culture medium inoculated.
C, condition of culture: move on to culturing room after the inoculation and cultivate, culturing room's temperature is 22～28 ℃, and culturing room's relative moisture is 30～40%, illumination every day 10～12 hours, and intensity of illumination is 1000～2000lx; The explant base portion of cultivating on about 25～32 days three kinds of different medium produces callus, and axil is sprouted 1～2 lateral bud; Cultivated 25～30 days again, and grew to the high bud of 2～3cm, the lateral bud base portion produces 2～5 adventive root, forms whole plant.
D, tissue cultivating seedling subculture: will constantly replace cultivation with II number with the III medium with the explant of callus and bud, the bud seedling of the more stalwartnesses of differentiation induces sturdy adventive root to transplant simultaneously.
The present invention has following advantage and effect:
1, utilize aseptic seedling to be explant, good stability, easy and simple to handle is drawn materials easily, need not pass through external sterilization, and convalescence is short; So far having one piece of report to adopt aseptic leaf, petiole and stem section is explant, but just grows callus after 45 days, changes the differential medium cultivation again over to and just can differentiate budlet point (yellow Jun plum etc., the torrid zone, Fujian science and technology, 2002,27 (1): 12～13) after 30 days.Induce callus and lateral bud synchronously about 25-32 of the present invention days.
2, cultivate by above-mentioned condition of culture, continue to cultivate 25～30 days, when bud 2～3cm was high, base portion produced 2～5 adventive root, formed to whole plant from going out more, only needed 50～62 days.Do sth. in advance 30～60 days formation whole plants than the present report shortest time and (seen blue celery English etc., gardening journal, 2003,30 (1): 107～109).
3, utilize two kinds of medium advantage separately alternately to cultivate, obtain the optimum efficiency that tissue of anthurium is cultivated, do not appearing in the newspapers at present.
4, explant all can produce callus, lateral bud, adventive root synchronously on I, II, III medium, and respectively has superiority; Yet there are no the report medium of callus induction, lateral bud, adventive root synchronously.
5, this technology path of---callus, bud, adventive root---whole plant that utilizes explant only needed 50～62 days to form whole plant, can obtain a large amount of whole plants in 4～5 months, carried out large-scale industrialized breeding.
Below in conjunction with embodiment optimum implementation of the present invention is described:
1, select 2～3cm height that tissue culture obtains and robust growth, even, the colory aseptic seedling of growing way for use, excision blade and petiole are cut into the stem section with 1～2 joint then earlier, are seeded on I, II, the III medium every bottle of 10～12 explants.
2, move on to culturing room after the inoculation and cultivate, culturing room's temperature is 22～28 ℃, and culturing room's relative moisture is 30～40%, illumination every day 12 hours, and intensity of illumination is 1000～2000lx;
3, cultivated about 30 days, average healing rate reaches 93%, and most of axil is sprouted 1-2 lateral bud (seeing Table 1).
The different medium of table 1 are spent the influence that the aseptic seedling callus forms and bud breaks up to the peace ancestral
The outer callus axil that minimal medium hormone (mg/L) inoculation outer planting produces callus group generation callus grows sprouting of lateral bud
The fate that body number (piece) is knitted is planted the lateral bud digit rate (%) of body number (piece) inductivity
(my god) (%)
I、1/2MS 6BA1.0+NAA0.05 100 29 84 84％ 152 89％
II、1/2MS 6BA2.0+NAA0.05 100 25 100 100％ 135 85％
III、1/2MS 6BA1.0+NAA0.05 100 32 95 95％ 194 100％
+IAA0.05+100g/LCM
(annotate: data are for cultivating the result of 35 days statistics in the table.)
4, the explant of above-mentioned cultivation continues to be seeded on three kinds of medium and cultivates, and cultivates 25～30 days again, and when growing to the high bud of 2～3cm, base portion grows 2～5 adventive root, forms whole plant (seeing Table 2).
The different medium propagation of table 2 are spent the influence of each organ breeding to the peace ancestral
Medium callus situation bud seedling differentiation situation number of seedling and growing way root situation blade situation
The I bulb is irregular, and most of terminal bud of differentiation and lateral bud aerial root are more, less on the about 1 part callus of diameter, about 0.5cm
2,
Centimetre, light green or yellowish sprouting are a little sprouted exhibition leaf, every callus 3-5 bar, 2-3cm light green for 3-5.
Look, the quality densification.It is long that tissue has into seedling 2-3.
Strain, high 1.5-3cm.
The II bulbous shape is obvious, and it is few and lack that the most of callus of diameter breaks up most of terminal bud and lateral bud aerial root, less, about 0.5cm
2,
About 2.5cm, pale green, yellowish sprouting a little, the more sprouting exhibition of number leaf, every callus 1-2 bar, 0.5-1.5cm light green.
Or yellow, the quality densification.(5-10).It is long that tissue has into seedling 1-2.
Strain, high 1-2cm.
The III bulbous shape is not obvious, and straight most of callus breaks up most of terminal bud and lateral bud 2-4 bar, and 1-3cm is long.Bigger, about 1cm
2,
The about 2cm in footpath, pale green, yellowish sprouting a little, the more sprouting exhibition of number leaf, every callus strong green, smooth.
Or yellow, quality is smooth to be caused (5-6).Tissue has into seedling 4-5
Close.Strain, high 2-3cm is sturdy.
(annotate: data are for cultivating 62 days statisticses in the table)
5, as can be seen: I medium callus is little and irregular, and differentiation bud point is few, and bud seedling growing way is general, grows but aerial root from table 1, table 2; Callus is big and shape is obvious on the II medium, and differentiation bud point is many, but a little less than the Cheng Miao and lack, and aerial root is few and lack; III medium callus is less, but the differentiation bud counts often, and more than the Cheng Miao and strengthen, aerial root is Duoed than the II medium and grown.Therefore explant is seeded in earlier on the II medium, changes the III medium then over to, changes the II medium again over to and replaces cultivation, and bud seedling, the sturdy adventive root of the more stalwartnesses of differentiation can cut transplanting.
2.2 the peace ancestral spends the acclimatization and transplants technology
An Zuhua likes higher temperature, and the growth thermophilic is 20～30 ℃, and more Xia Wendu is not higher than 31 ℃, and the temperature of surviving the winter is not less than 10 ℃, like gentle, so every day need to foliar spray.Yet test-tube plantlet all is under artificial condition in the bottle seedling stage, various optimum growth conditionss are provided, and finishes process of growth; Therefore, widely different with extraneous natural environmental condition, can adapt to extraneous natural environmental condition in order to make bottle seedling, just must allow test-tube plantlet that the procedure of adaptation of a transition, i.e. hardening are arranged.
Most of plant can constantly grow, breed in the test tube breeding, but does not take root and can't transplant; Tissue of anthurium culture technique reported method is to take root for transplanting through 25～40 days for (seeing yellow Jun plum etc., the torrid zone, Fujian science and technology, 2002,27 (1): 12～13 mostly again both at home and abroad; Cai Weifan, subtropical plant science, 2002,31 (3): 66-68 etc.).
During test-tube seedling transplanting, because of organizing the children tender, be easy to grow assorted bacterium, and the peace ancestral spends root, stem, the Ye Junhui morbidity of test-tube plantlet, the plant that is injured during morbidity at first stops growing, and the position that is injured then is filbert, and the later stage stem is covered with white or the mould layer of pink, last complete stool withered death has reduced transplanting survival rate.
The objective of the invention is the deficiency at above technology, invent a kind of cultural method that lures bud and root synchronously, the cutting test seedling strengthening seedling and rooting just can form the full seedling of root leaf tool; And with different medicament immersion treatment, reduce the germ source when transplanting, pre-disease prevention is accelerated growth.
The invention provides a kind of peace ancestral and spend seedling exercising replanting technique for test-tube plantlet, mainly carry out with method according to the following steps:
A, select for use II, III medium alternately to cultivate the strong sprout that is obtained, need not to go to root media, can form whole plant; Therefore in incubation along with the appearance of adventive root, can transplant by bottle outlet, simplify the breeding program.
B, transplanting preliminary treatment: soaked 5～25 minutes with medicaments such as 500～2000 times potassium permanganate, carbendazim, mancozeb, thiophanate methyl, family expenses streptomycins.
C, cultivation matrix: select the coconut palm chaff during transplanting for use: peat soil: perlite=2: 2: 1 is matrix; Use before the transplanting and soak medicament with concentration and drench and placed 2～3 days, and then transplant.
D, the management of transplanting back: after the transplanting, change seeding room over to, carry out hardening with diaphragm seal, temperature is controlled at 22～28 ℃, and humidity 60～80% was sprayed water once in film, and slowly opened film after 20～40 days in 2～7 days; Transplanted back 10 days, and began when young leaves grows, to be transferred to hot-house culture with MS mother liquor spraying (once in a week).
The present invention has following advantage and effect:
1, adopt medium alternately to cultivate and lure bud and root synchronously, strengthening seedling and rooting just can form the full seedling of root leaf tool separately, grow to 2～3cm high the time, can cut transplanting; Callus and weak seedling can continue to change over to medium and continue to cultivate, and the method is simplified the breeding program.
When 2, transplanting, all with medicament disinfections of seedling and cultivation matrix reduce the germ source, build up resistance, so improve survival rate, quickening growth rate.
3, select for use loosely ventilative, fertile, be rich in organic matter, the crude culture matrix of no damage by disease and insect is used before the transplanting and is soaked medicament with concentration and drench and placed soil humidity about 60% when guaranteeing transplanting 2～3 days.
4, in indoor hardening, dwindle changes in environmental conditions, realize that height survives, transplanting method cheaply.
Below in conjunction with embodiment optimum implementation of the present invention is described:
1, cut down growing to the high seedling of 2～3cm, then seedling is put into clear water the agar of root is cleaned up, note water temperature when washing seedling, should use warm water winter, in order to avoid water temperature is crossed low frostbite seedling.
2, test-tube plantlet is cleaned the back and is soaked processing in 5～25 minutes with medicaments such as 500～2000 times potassium permanganate, carbendazim, mancozeb, thiophanate methyl, family expenses streptomycins.
3, with the coconut palm chaff: peat soil: the ratio preparation matrix of perlite=2: 2: 1 guarantees that the porosity and the water retention capacity of matrix ventilation was good with 1: 1; Use before the transplanting and soak medicament with concentration and drench and placed soil humidity about 60% when guaranteeing to transplant 2～3 days.
When 4, transplanting, can not directly directly insert the seedling of band root in the matrix, fracture easily, cause wound infection because of the seedling root is more crisp.With small wooden or tweezers the matrix of cave dish is dug a duck eye earlier, seedling is put into gently covers matrix again root covered; Firmly do not push matrix, in order to avoid matrix is real excessively, root system poor aeration and cause rotting of root.
5, after the transplanting, in seeding room, use the diaphragm seal hardening, temperature is controlled at 22～28 ℃, humidity 60～80%; In film, sprayed water once, and slowly opened film after 20～40 days in 2～7 days; Transplant after 10 days, begin when young leaves grows, to be transferred to the greenhouse and to cultivate with MS mother liquor spraying (once in a week).
Adopt above method to transplant, the plant strain growth stalwartness, fast growth, the rate of catching an illness is low, and transplanting survival rate reaches more than 95%.
Claims (2)
1, a kind of peace ancestral spends plant tissue's culture technique, it is characterized in that pacifying the ancestral, to spend the aseptic seedling tissue culture technique be to carry out with method according to the following steps: the selection of A, explant: selecting for use the peace ancestral to spend 2～3cm height that young tender organ obtains through tissue culture and robust growth, growing way is even, quality is good aseptic seedling is material, and the stem section that cuts with 1～2 joint is that explant is cultivated.
The inoculation of B, explant: will be with the stem section of 1～2 joint, be seeded in I.1/2MS+6-BA0.5～1.0mg/L+NAA0.05mg/L+3% glucose+0.75% agar, II.1/2MS+6-BA1.5～2.0mg/L+NAA0.05mg/L+3% glucose+0.75% agar, III.1/2MS+6-BA1.0～1.5mg/L+NAA0.05mg/L+IAA0.05mg/L+3% glucose+0.75% agar, 10～12 stem sections of every bottle of culture medium inoculated.
C, condition of culture: move on to culturing room after the inoculation and cultivate, culturing room's temperature is 22～28 ℃, and culturing room's relative moisture is 30～40%, illumination every day 10～12 hours, and intensity of illumination is 1000～2000lx; Cultivated about 25～32 days, the explant base portion on three kinds of different medium produces callus, and axil is sprouted 1～2 lateral bud; Continue to cultivate 25～30 days, grow to the high bud of 2～3cm, the lateral bud base portion produces 2～5 adventive root, forms whole plant.
D, tissue cultivating seedling subculture: will constantly replace cultivation with II number with the III medium with the explant of callus and budlet, the bud seedling of the more stalwartnesses of differentiation induces sturdy adventive root to transplant simultaneously.
2, a kind of tissue of anthurium culture technique, it is characterized in that pacifying the ancestral, to spend seedling exercising replanting technique for test-tube plantlet be to carry out with method according to the following steps:
A, select for use II, III medium alternately to cultivate the strong sprout that is obtained, need not to go to root media, can form whole plant; Therefore in incubation along with the appearance of adventive root, can transplant by bottle outlet, simplify the breeding program.
B, transplanting preliminary treatment: soaked 5-25 minute with medicaments such as 500-2000 times potassium permanganate, carbendazim, mancozeb, thiophanate methyl, family expenses streptomycins.
C, cultivation matrix select: select the coconut palm chaff during transplanting for use: peat soil: perlite=2: 2: 1 is matrix; Use before the transplanting and soak medicament with concentration and drench and placed 2-3 days, and then transplant.
D, the management of transplanting back: after the transplanting, change seeding room over to, use the diaphragm seal hardening, temperature is controlled at 22～28 ℃, humidity 60～80%; In film, sprayed water once, and slowly opened film after 20～40 days in 2～7 days; Transplanted back 10 days, and began when young leaves grows, to be transferred to hot-house culture with MS mother liquor spraying (once in a week).
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| CN103734009A (en) * | 2013-12-20 | 2014-04-23 | 三明市农业科学研究院 | Tissue culture method of anthurium |
| CN104642108A (en) * | 2015-02-03 | 2015-05-27 | 中国科学院亚热带农业生态研究所 | Method suitable for tissue culture mass production of multiple plants |
| CN105010141A (en) * | 2015-07-09 | 2015-11-04 | 虞龙 | Anthurium rapid propagation method |
| CN105493951A (en) * | 2015-12-30 | 2016-04-20 | 广西壮族自治区农业科学院生物技术研究所 | Transplanting method of tissue-culture derived banana plantlets |
| CN105519418A (en) * | 2015-12-30 | 2016-04-27 | 广西壮族自治区农业科学院生物技术研究所 | Method for transplanting passiflora edulis tissue cultured seedlings |
| CN105532402A (en) * | 2015-12-30 | 2016-05-04 | 广西壮族自治区农业科学院生物技术研究所 | Transplanting method of siraitia grosvenorii tissue culture seedlings |
| CN105638405A (en) * | 2015-12-30 | 2016-06-08 | 广西壮族自治区农业科学院生物技术研究所 | Papaya tissue culture seedling transplanting method |
| CN111084087A (en) * | 2019-12-04 | 2020-05-01 | 华南农业大学 | Method for inhibiting 'Pinguanjun anthurium' cluster seedlings from happening through root cutting and cutting cultivation |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1353926A (en) * | 2000-11-20 | 2002-06-19 | 深圳市金百合生物工程技术有限公司 | Red palm tissue plant culture technique |
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| CN103734009A (en) * | 2013-12-20 | 2014-04-23 | 三明市农业科学研究院 | Tissue culture method of anthurium |
| CN103734009B (en) * | 2013-12-20 | 2015-06-17 | 三明市农业科学研究院 | Tissue culture method of anthurium |
| CN104642108A (en) * | 2015-02-03 | 2015-05-27 | 中国科学院亚热带农业生态研究所 | Method suitable for tissue culture mass production of multiple plants |
| CN105010141A (en) * | 2015-07-09 | 2015-11-04 | 虞龙 | Anthurium rapid propagation method |
| CN105493951A (en) * | 2015-12-30 | 2016-04-20 | 广西壮族自治区农业科学院生物技术研究所 | Transplanting method of tissue-culture derived banana plantlets |
| CN105519418A (en) * | 2015-12-30 | 2016-04-27 | 广西壮族自治区农业科学院生物技术研究所 | Method for transplanting passiflora edulis tissue cultured seedlings |
| CN105532402A (en) * | 2015-12-30 | 2016-05-04 | 广西壮族自治区农业科学院生物技术研究所 | Transplanting method of siraitia grosvenorii tissue culture seedlings |
| CN105638405A (en) * | 2015-12-30 | 2016-06-08 | 广西壮族自治区农业科学院生物技术研究所 | Papaya tissue culture seedling transplanting method |
| CN111084087A (en) * | 2019-12-04 | 2020-05-01 | 华南农业大学 | Method for inhibiting 'Pinguanjun anthurium' cluster seedlings from happening through root cutting and cutting cultivation |
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