CN100424169C - Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method - Google Patents
Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method Download PDFInfo
- Publication number
- CN100424169C CN100424169C CNB2005100050177A CN200510005017A CN100424169C CN 100424169 C CN100424169 C CN 100424169C CN B2005100050177 A CNB2005100050177 A CN B2005100050177A CN 200510005017 A CN200510005017 A CN 200510005017A CN 100424169 C CN100424169 C CN 100424169C
- Authority
- CN
- China
- Prior art keywords
- callus
- composition
- breeding
- culture
- tissue culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses a culture medium for anthurium andraeanum tissue culture and a method for breeding tissue culture seedlings. The concentration combination of major element components in the culture medium does not exist in the existing culture mediums for anthurium andraeanum tissue culture, and simultaneously, abscisic acid used in the culture medium is not used in the prior art for fast breeding anthurium andraeanum. The method for breeding tissue culture seedlings of anthurium andraeanum comprises the steps that: leaves or leaf stalks are induced to generate calluses and adventitious buds by utilizing the culture medium; the calluses which polarize out the adventitious buds are transplanted onto an improved MS culture medium to culture and to breed adventitious branches; the adventitious branches are transplanted into a rooting culture medium to be induced to root, and then, the rooting tissue culture seedlings are transplanted into soil substrate by using a conventional transplantation method for the tissue culture seedlings. By adopting the technical scheme provided by the present invention, the calluses and regenerated plants of the leaves and the leaf stalks of nutrition-breeding potted seedlings of each of two varieties of anthurium andraeanum can be induced, the influence of variety genotypes on the induction of the calluses of the leaves and the leaf stalks and the plantlets regeneration is reduced, and the breeding speed of the tissue culture seedlings of anthurium andraeanum is quickened.
Description
Technical field
The present invention relates to tissue cultured seedling in the Plant Biotechnology (test-tube plantlet) breeding technology, spend the substratum and the tissue cultured seedling propagating method of group training usefulness in particular to a kind of ancestral of peace.
Background technology
An Zuhua (claiming the red palm, fancy candles lit in the bridal chamber at wedding, fiery crane again) is Araeceae (Araceae) peace ancestral Pittosporum (Anthurium) plant.Main commodity flower cultivar has A.andreanum and A.scherzerianum, and its sales volume occupies the second on flowers market, the world, be to be only second to the blue rare flower in the torrid zone.The breeding of peace ancestral flower seedling is mainly by the breeding of tissue cultured seedling cultured method.Since (1974) such as Pierik induced A.andreanum to form callus, (2004) such as (2003), Martin etc. (2003), Vargas such as (1997), Xiao Sanyuan and Liang Guoping (2000), Joseph such as Pierik (1976), Kunisaki (1980), Kuehnle and Sugii (1991), Teng (1997), Chen etc. (2004) and Guo Weiming had reported the tissue cultured seedling breeding of A.andreanum potted flower and cut-flower respectively; Geier (1986), Liu Chunming and Xu Zhihong (1992) and Hamidah etc. (1997) have reported that respectively the A.scherzerianum organ takes place and somatic cell embryogenesis regeneration plant.The characteristics of prior art show following several respects.
When (1) breeding A.andreanum and A.scherzerianum fast, use the substratum that has nothing in common with each other
Induce A.scherzerianum callus and plant regeneration then to adopt Nitsch (Nitsch, 1969) minimum medium or partly measure MS (Murashige and Skoog, 1962) substratum and reduce NH
4NO
3Content.For example, Geier (1986) induces the A.scherzerianum callus to use the Nitsch minimum medium, reduces NH
4NO
3Concentration is added 1mg/L6-benzyladenine and 0.1mg/L 2,4 dichlorophenoxyacetic acid; Induce plant regeneration to use identical minimum medium, improve NH with breeding
4NO
3Concentration is added the 0.5mg/L6-benzyladenine; Root media is identical with regeneration and propagating culture medium, but does not add growth regulator.
The substratum of inducing A.andreanum leaf and petiole explant callus and plant regeneration to use is MS minimum medium (Kunisaki, 1980; Teng, 1997) and MS macroelement reduce by half (Kuehnle and Sugii, 1991; Chen etc., 1997; Martin etc., 2003; Joseph etc., 2003) or the reduce by half substratum of (Kuehnle and Sugii, 1991) of part macroelement.Martin etc. (2003) all use and partly measure the MS substratum in A.andreanum cutting flower variety plant regeneration process, growth regulator evoked callus, the indefinite bud by adding different sorts and different concns and taking root.Teng (1997), is taken root on the substratum of no growth regulator containing evoking adventive bud on the MS substratum of 6-benzyl aminopurine at MS substratum (adding 2.2-4.4 μ M 6-benzyl aminopurine and 0.9 μ M 2,4 dichlorophenoxyacetic acid) evoked callus.
So far, do not see as yet about a kind of minimum medium all be fit to the peace ancestral spend the callus induction of 2 kinds and the report of plant regeneration.
(2) reproducible kind of prior art or genotype are limited
Many potted flowers and cutting flower variety in A.andreanum and A.scherzerianum, have been cultivated respectively.In the research of same kind, Kunisaki (1980) has reported the quick breeding of 4 kinds, Geier (1986) only induces 10 genotype regeneration plants from 18 genotype, Kuehnle and Sugii (1991) use leaf explant callus and the regeneration plant that same medium is induced 7 kinds, Liu Chunming and Xu Zhihong (1992) have been induced the genotype plant regeneration of 7 seed sources, Chen etc. (1997) induce the root explant regeneration plant of 2 kinds, (2003) such as Joseph (2003) and Martin induce the leaf explant regeneration plant of 3 and 2 cutting flower varieties respectively, (2004) such as Vargas etc. (2004) and Guo Weiming have reported the plant regeneration of a kind respectively, and kind quantity is not reported in other researchs.These research report explanations utilize existing substratum can only induce the callus and the regeneration plant of indivedual kinds, do not develop the substratum of peace ancestral's flower variety or ind callus induction of genotype and plant regeneration as yet.
The potted plant seedling leaf of (3) nourishing and generating and the breeding technology of petiole explant regeneration plant are immature
The explant that is used for evoked callus and regeneration plant has embryo, vegetative bud, leaf, petiole, spadix, spathe, root and stem section.Wherein, leaf and petiole explant research are many, and 80% research uses leaf and petiole to make explant.The peace ancestral spends and induces leaf and the general approach that adopts of petiole regeneration plant to be in the industry, at first obtains the aseptic seedling of seed germination or cultivates the generation tissue cultured seedling by stem eye, and then induce tissue cultured seedling leaf and petiole to produce callus and regeneration plant.The research that has is explant (Kunisaki, 1980 with seedling or tissue culture seeding stem segment, leaf and the petiole of seed germination also; Geier, 1986; Liu Chunming and Xu Zhihong, 1992; Hamidah etc., 1997; Guo Weiming etc., 2004; Vargas etc., 2004).
Another kind of approach is directly to induce potted plant seedling leaf and petiole to produce callus and regeneration plant (Teng, 1997; Xiao Sanyuan and Liang Guoping, 2000; Joseph, 2003; Martin etc., 2003).Induce difficulty or ease with regard to callus and regeneration plant, relatively easy is the leaf and the petiole explant of seedling and test-tube plantlet, and the potted plant seedling explant difficulty of nourishing and generating is big.The former can induce more kind leaf explant regeneration plant, and the latter has 3 kinds at most by regeneration induction plant (Joseph, 2003).Is that the quantity of drawing materials is big and the time of drawing materials is long, helps introducing a fine variety promoting and breed fast new variety with the potted plant seedling leaf of nourishing and generating and petiole as the advantage of explant.The quick breeding current techique of therefore, the research and development peace ancestral seeds of flowering plants and potted plant seedling leaf of kind and petiole regeneration plant has very big using value.Existing research is only succeedd on indivedual kinds, has limited the practical application of result of study.
Summary of the invention
Technical problem to be solved by this invention is to overcome above-mentioned deficiency of the prior art, spend group to train the substratum and the tissue cultured seedling propagating method of usefulness and propose a kind of ancestral of peace, use this substratum and method breeding peace ancestral to spend tissue cultured seedling, potted plant seedling leaf and petiole callus and the regeneration plant that can induce the peace ancestral to spend 2 kind A.andreanum and each kind of A.scherzerianum to nourish and generate, alleviate the influence of kind or genotype, and can accelerate to pacify the reproduction speed that the ancestral spends tissue cultured seedling leaf and petiole callus induction and plant regeneration.
Technical scheme provided by the present invention is:
A kind of ancestral of peace spends the substratum of group training usefulness, is used to induce leaf and petiole callus and differentiation adventitious buds thereof, comprises following ingredients:
Prescription 1:
(1) macroelement composition
KNO
3，400～850mg/L；NH
4NO
3，150～350mg/L；CaCl
2·2H
2O，200～500mg/L；MgSO
4·7H
2O，150～400mg/L；KH
2PO
4，100～250mg/L；
(2) micro-composition
FeSO
4·7H
2O，13.9～27.8mg/L；Na
2EDTA·2H
2O，18.65～37.3mg/L；KI，0～0.83mg/L；H
3BO
3，0～6.2mg/L；MnSO
4·4H
2O，11.2～25mg/L；ZnSO
4·7H
2O，4.3～10mg/L；Na
2MoO
4·2H
2O，0.25mg/L；CuSO
4·5H
2O，0.01～0.025mg/L；CoCl
2·6H
2O，0.025mg/L；
(3) organotrophy composition
Inositol, 500～1000mg/L; Nicotinic acid, 0.5～5mg/L; Pyridoxine hydrochloride, 0.5～1mg/L; Vitamin, 0.1～1mg/L; Glycine, 2.0mg/L;
(4) growth regulator
2,4 dichlorophenoxyacetic acid, 0.1～0.5mg/L; 6-benzyl aminopurine, 0.5～1.5mg/L; Dormin 0.01～0.3mg/L;
(5) carbon source: sucrose, 20～30g/L;
(6) peptizer: agar, 6～7g/L;
Final pH is adjusted to 5.6～5.8;
Prescription 2:
(1) macroelement composition
KNO
3，400～850mg/L；NH
4NO
3，150～350mg/L；CaCl
2·2H
2O，200～500mg/L；MgSO
4·7H
2O，150～400mg/L；KH
2PO
4，100～250mg/L；
(2) micro-composition
FeSO
4·7H
2O，13.9～27.8mg/L；Na
2EDTA·2H
2O，18.65～37.3mg/L；KI，0～0.83mg/L；H
3BO
3，0～6.2mg/L；MnSO
4·4H
2O，11.2～25mg/L；ZnSO
4·7H
2O，4.3～10mg/L；Na
2MoO
4·2H
2O，0.25mg/L；CuSO
4·5H
2O，0.01～0.025mg/L；CoCl
2·6H
2O，0.025mg/L；
(3) organotrophy composition
Inositol, 500～1000mg/L; Nicotinic acid, 0.5～5mg/L; Pyridoxine hydrochloride, 0.5～1mg/L; Vitamin, 0.1～1mg/L; Glycine, 2.0mg/L;
(4) growth regulator
α-Nai Yisuan 0.1～0.5mg/L, 6-benzyl aminopurine 0.5～1.5mg/L;
(5) carbon source: sucrose, 20～30g/L;
(6) peptizer: agar, 6～7g/L;
Final pH is adjusted to 5.6～5.8.
A kind of peace ancestral spends tissue cultured seedling propagating method, comprises the steps:
(1) induce leaf or petiole to produce callus and indefinite bud
Utilize the above-mentioned prescription 1 or the substratum described in 2 of filling a prescription to induce and produce callus and indefinite bud: the potted plant seedling leaf that will nourish and generate or petiole are seeded on the substratum that adopts prescription 1 preparation, after cultivating for 3～8 weeks, induce the generation callus, or then induce the generation indefinite bud, though or generation callus, when not producing indefinite bud, then will be greater than 3～5mm
3Callus transfer on the substratum that adopt prescription 2 preparations, after cultivating through 6～8 weeks, callus is by the regeneration induction indefinite bud again;
(2) adventitious shoot breeding
Above-mentioned callus with differentiation capability has promptly been differentiated the callus of indefinite bud, and subculture is cultivated on modified MS medium, propagation callus and breeding tissue cultured seedling fast; Callus is cultivated in 1: 4 ratio shoot proliferation after cultivating for 6～7 weeks on the modified MS medium; When being used for tissue cultured seedling and breeding fast, the callus subculture is in 1: 2 ratio subculture, and keeps the adventitious shoot less than 0.5cm length on callus;
(3) tissue cultured seedling root induction and transplanting
With length is that adventitious shoot more than 2～3cm is transferred in the root media, and after cultivating through 2～4 weeks, adventitious shoot produces new root, and new root growth to 0.5～when 1cm is long adopts the conventional tissue cultured seedling method for transplanting tissue cultured seedling of will taking root to be transplanted in the soil matrix.
Preferably, the macroelement composition of described inducing culture is KNO
3, 600mg/L; NH
4NO
3, 200mg/L; CaCl
22H
2O, 220mg/L; MgSO
47H
2O, 370mg/L; KH
2PO
4, 170mg/L.
Further, described peace ancestral spends tissue culture propagation method, wherein the 2nd goes on foot used modified MS medium, and the growth regulator composition in this substratum is: α-Nai Yisuan, 0.1～0.5mg/L, 6-benzyl aminopurine, 0.4～2.0mg/L; Carbon source: sucrose, 20～30g/L; Peptizer: agar, 4～6g/L; Macroelement composition, micro-composition, organotrophy composition are identical with corresponding composition in the MS minimum medium; Final pH is adjusted to 5.6～5.8.
Further, described peace ancestral spends tissue culture propagation method, and wherein used root media of the 3rd step contains the MS minimum medium composition that macroelement content reduces by half in this substratum; Naphthylacetic acid, 0.05～0.2mg/L; Sucrose, 10g/L; Agar, 7g/L; The final pH value is adjusted to 5.6～5.8.
The present invention has following advantage: (1) has been set up new peace ancestral's flowerpot and has been planted Miao Ye and petiole callus inducing medium, do not have in the existing substratum of the concentration combination of the macroelement composition in this substratum, simultaneously the dormin that uses in this substratum be existing peace ancestral spend do not have in the quick breeding technology used, combination of macroelement composition and dormin are the key factors of inducing leaf and the success of petiole callus, under the suitable situation of other conditions, lack any processing in above-mentioned two kinds of processing, callus induction is suppressed.Inositol concentration in the substratum is brought up to 500～1000mg/L, help inducing of callus.(2) set up the peace ancestral and spent the system of the quick breeding of tissue cultured seedling.This individual system is made up of the inducing culture of setting up and improvement inducing culture, modified MS medium and 1/2MS (macroelement concentration reduces by half) root media.Inducing culture is the basis of this rapid propagation system, have only and on inducing culture or improvement inducing culture, to have the callus succeeding transfer culture of differentiation capability on modified MS medium, could promote the frequency of callus propagation and raising differentiation adventitious buds, promote the peace ancestral to spend the reproduction speed of tissue cultured seedling.(3) set up the peace ancestral and spent 2 kind A.andreanum and the general quick breeding technology of tissue cultured seedling of A.scherzerianum, can induce the peace ancestral to spend the leaf and the petiole callus of the potted plant seedling that 2 kind A.andreanum and A.scherzerianum nourish and generate, and regeneration plant, alleviated the influence of kind or genotype to leaf and petiole callus induction and plant regeneration.Utilize technical solution of the present invention can induce potted plant seedling leaf of each kind of An Zuhua and petiole to produce callus and regeneration plant.
Embodiment
At first preparation breeding peace ancestral spends the required substratum of tissue cultured seedling, and conventional substratum compound method is adopted in the making of substratum.Callus induction and differentiation adventitious buds substratum adopt inducing culture, and the prescription of this inducing culture is:
Prescription 1:
Macroelement composition: KNO
3, 600mg/L; NH
4NO
3, 200mg/L; CaCl
22H
2O, 220mg/L; MgSO
47H
2O, 370mg/L; KH
2PO
4, 170mg/L;
Trace element composition: FeSO
47H
2O, 13.9mg/L; Na
2EDTA2H
2O, 18.65mg/L; KI, 0.83mg/L; H
3BO
3, 6.2mg/L; MnSO
44H
2O, 22.3mg/L; ZnSO
47H
2O, 8.6mg/L; Na
2MoO
42H
2O, 0.25mg/L; CuSO
45H
2O, 0.025mg/L; CoCl
26H
2O, 0.025mg/L;
Organotrophy composition: inositol, 1000mg/L; Nicotinic acid, 0.5mg/L; Pyridoxine hydrochloride, 0.5mg/L; Vitamin, 0.1mg/L; Glycine, 2.0mg/L;
Growth regulator: 2,4 dichlorophenoxyacetic acid, 0.2mg/L; 6-benzyl aminopurine, 1.0mg/L; Dormin, 0.1mg/L;
Carbon source: sucrose, 20g/L;
Peptizer: agar, 6g/L;
Final pH is adjusted to 5.6.
Among the present invention this substratum is called the Xiao substratum.
Prescription 2:
Macroelement composition: KNO
3, 600mg/L; NH
4NO
3, 200mg/L; CaCl
22H
2O, 220mg/L; MgSO
47H
2O, 370mg/L; KH
2PO
4, 170mg/L;
Trace element composition: FeSO
47H
2O, 13.9mg/L; Na
2EDTA2H
2O, 18.65mg/L; KI, 0.83mg/L; H
3BO
3, 6.2mg/L; MnSO
44H
2O, 22.3mg/L; ZnSO
47H
2O, 8.6mg/L; Na
2MoO
42H
2O, 0.25mg/L; CuSO
45H
2O, 0.025mg/L; CoCl
26H
2O, 0.025mg/L;
Organotrophy composition: inositol, 1000mg/L; Nicotinic acid, 0.5mg/L; Pyridoxine hydrochloride, 0.5mg/L; Vitamin, 0.1mg/L; Glycine, 2.0mg/L;
Growth regulator: α-Nai Yisuan, 0.1mg/L, 6-benzyl aminopurine, 1.0mg/L,
Carbon source: sucrose, 20g/L;
Peptizer: agar, 6g/L;
Final pH is adjusted to 5.6.
Among the present invention this substratum is called improvement Xiao substratum.
The substratum of adventitious shoot breeding adopts modified MS medium, and it consists of:
Macroelement composition: KNO
3, 1900mg/L; NH
4NO
3, 1650mg/L; CaCl
22H
2O, 440mg/L; MgSO
47H
2O, 370mg/L; KH
2PO
4, 170mg/L;
Trace element composition: FeSO
47H
2O, 27.8mg/L; Na
2EDTA2H
2O, 37.3mg/L; KI, 0.83mg/L; H
3BO
3, 6.2mg/L; MnSO
44H
2O, 22.3mg/L; ZnSO
47H
2O, 8.6mg/L; Na
2MoO
42H
2O, 0.25mg/L; CuSO
45H
2O, 0.025mg/L; CoCl
26H
2O, 0.025mg/L;
Organotrophy composition: inositol, 100mg/L; Nicotinic acid, 0.5mg/L; Pyridoxine hydrochloride, 0.5mg/L; Vitamin, 0.1mg/L; Glycine, 2.0mg/L;
Growth regulator: α-Nai Yisuan, 0.1mg/L, 6-benzyl aminopurine, 1.0mg/L;
Carbon source: sucrose, 20g/L;
Peptizer: agar, 5g/L;
Final pH is adjusted to 5.7;
Consisting of of root media: MS minimum medium (macroelement content reduces by half)+0.1mg/L α-Nai Yisuan+10g/L sucrose+0.7% agar, pH5.8, that is:
Macroelement composition: KNO
3, 950mg/L; NH
4NO
3, 825mg/L; CaCl
22H
2O, 220mg/L; MgSO
47H
2O, 185mg/L; KH
2PO
4, 85mg/L;
Trace element composition: FeSO
47H
2O, 27.8mg/L; Na
2EDTA2H
2O, 37.3mg/L; KI, 0.83mg/L; H
3BO
3, 6.2mg/L; MnSO
44H
2O, 22.3mg/L; ZnSO
47H
2O, 8.6mg/L; Na
2MoO
42H
2O, 0.25mg/L; CuSO
45H
2O, 0.025mg/L; CoCl
26H
2O, 0.025mg/L;
Organotrophy composition: inositol, 100mg/L; Nicotinic acid, 0.5mg/L; Pyridoxine hydrochloride, 0.5mg/L; Vitamin, 0.1mg/L; Glycine, 2.0mg/L;
Growth regulator: α-Nai Yisuan, 0.1mg/L;
Carbon source: sucrose, 10g/L;
Peptizer: agar, 7g/L;
Final pH is adjusted to 5.8.
Macroelement composition in the substratum, ferrous components, other micro-compositions and organotrophy composition are mixed with stock solution by 10 times, 200 times, 1000 times and 100 times of concentration respectively, simultaneously required growth regulator is mixed with the mother liquor of 1mg/ml, is kept in 4 ℃ of refrigerators.Amount is prepared each substratum and is regulated pH with 1N NaOH on demand then, boils the back branch and installs in the culturing bottle, and 121 ℃ of sterilizations 15 minutes, the cooling back was standby.
The clip petiole is about 5cm, does not launch blade, and the flowing water flushing with clean filter paper suck dry moisture, was cut into suitable size with explant and places aseptic beaker after 30 minutes, carried out following surface sterilization in Bechtop.70% alcohol immersion 30 seconds is used 0.1%HgCl again
2Handled 6-8 minute, the sterilizing process discontinuous is shaken beaker.After the surface sterilization, clean explant 5 times with aseptic deionized water, each residence time is 3-5 minute, and shakes the thimerosal on the abundant eccysis explant of beaker.
Blade after the sterilization cleaning is cut into about 1cm along master pulse
2Size is cut into length about 1cm with petiole, is inoculated on the Xiao substratum, and every bottle (50ml) inoculates 3-4 explant.The explant of inoculation is placed on dark or the low light level, and (cultivate under 300～500lx) conditions, culture temperature is 25 ± 1 ℃.
After inoculating for 3 weeks, when the explant incision grows callus gradually, culture is transferred to cultivation under the illumination condition.Culture condition from now on is: intensity of illumination 4000lx, the photoperiod is irradiation 16 hours and dark 8 hours, 25 ± 1 ℃ of temperature.Callus is in Xiao substratum or continued growth and differentiate indefinite bud gradually behind subculture on the same substratum.For example, 3 weeks of Abbe Shen (Ambition) petiole inoculation back produce callus, 6～7 week of inoculation back differentiation indefinite bud.If callus growth to 3～5mm
3Size does not have differentiation adventitious buds, and callus need be transferred on the improvement Xiao substratum and cultivate for 6～8 weeks, the evoking adventive bud differentiation, culture condition is the same.For example, be inoculated in Ya Lisangna (Arizona) blade on the Xiao substratum induced produce callus after, need could be to the improvement Xiao substratum by the regeneration induction indefinite bud with the callus subculture.The blade callus induction rate is lower than petiole.To Ya Meige (Amigo), watt logical sequence sky slave (Valentino), red cutting, orange champion (Orange Champion, greatly), orange champion's (little), 7 kind blades of Ya Lisangna (Arizona) and sweetie (Sweetheart Red) and petiole callus induction rate on average are respectively 63.2% and 85.8%.
Behind the callus differentiation indefinite bud, this callus with differentiation capability is shifted subculture on modified MS medium, cultivate.Modified MS medium plays the function that promotes callus propagation and adventitious shoot breeding.Generally at first promote callus propagation, treatment process is the 6-benzyl aminopurine level that improves in the modified MS medium, makes callus growth fast, and differentiation adventitious buds is less relatively.When the callus amount acquires a certain degree, need to reduce the callus growth amount, promote differentiation adventitious buds and growth, treatment process is to reduce 6-benzyl aminopurine concentration in the MS improved culture medium.Any culturing purposes no matter, callus per 6～7 all subcultures 1 time.Wherein, in the callus breeding in 1: 4 ratio succeeding transfer culture, and in the adventitious shoot reproductive process in 1: 2 ratio succeeding transfer culture.As if after the indefinite bud on the callus is excised fully, evoked callus breaks up indefinite bud and grows into 2-3cm length again, needs to cultivate the time about 10 weeks during subculture.Therefore, fast the subculture callus need keep indefinite bud and less than the adventitious shoot of 3～5mm length in the reproductive process, after these indefinite buds and adventitious shoot are cultivated through 6～7 weeks again, just can grow into the above length of 2～3cm, guarantee that each subculture can both cut off the adventitious shoot of suitable height, is used for root induction.(each subculture can obtain 30～40 strain unrooted tissue cultured seedling and is used for root induction among the φ=8.8cm) at 1 big culturing bottle.
The adventitious shoot that 2～3cm is long cuts off from callus, is inoculated on the root media, through cultivating about 3 weeks, produces tender new of white children at basal part of stem, and every strain tissue cultured seedling can be induced 3～5 new roots.When new root growth to 5～10mm length, the tissue cultured seedling of will taking root is transplanted in the soil matrix.Soil matrix is made up of coir, vermiculite and perlite, and each accounting example is 1/3.The tissue cultured seedling of transplanting places the greenhouse to cultivate, and the transplanting initial stage hides with plastics film, and keeping relative air humidity is about 90%, 20～25 ℃ of temperature, intensity of illumination 3000～5000lx.When 2～3 weeks, the back tissue cultured seedling recovered growth, open plastics film gradually, relative air humidity remains on about 70% in the greenhouse, and improving illumination is about 8000lx.The tissue cultured seedling transplanting survival rate reaches more than 90%.Transplanted back about 4 months, tissue cultured seedling grows into 8～10cm height, tissue cultured seedling is entered the cultivation management of next stage with seedling as production.
Use the Xiao substratum, or cooperates with improvement Xiao substratum the existing potted plant seedling of introducing of all peace ancestral's flower varieties is all successfully induced leaf and petiole generation callus and regeneration plant, these variety names are listed below.
27 Cultivars are arranged in the A.andreanum kind, be Ya Meige (Amigo), watt logical sequence sky slave (Valentino), Atlanta (Atlanta), red cutting, Ka Xiluo (Casino), orange champion (OrangeChampion), orange champion (little), red roasting draw (Carre), Ya Lisangna (Arizona), Robinson, Crusoe (Robino), sweetie (Sweetheart Red), Abbe Shen (Ambition), powder champion (PinkChampion), Beijing success (Beijing success), tie up he (Vitara), holder card (Tucano), I clings to agate (Alabama), Mississippi (Mississippi), blessing (Impreza), auspicious horse (Rima), Dao Keta (Dakota), quiet (Silence), Sha Wode (Sharade), Minnesota (Minnesota), Latin America (Latino), crolla many (Colorado), Missouri (Missouri).
3 Cultivars are arranged, i.e. A Tusi (Artus), Santiago Solari (Solara), carat Promised Land (Graffity) in the A.scherzerianum kind.
Claims (3)
1. pacify the inducing culture that the ancestral spends group training usefulness for one kind, be used for evoked callus and regenerated adventitious bud, it is characterized in that comprising following ingredients:
Prescription 1:
(1) macroelement composition
KNO
3，400～850mg/L；NH
4NO
3，150～350mg/L；CaCl
2·2H
2O，200～500mg/L；MgSO
4·7H
2O，150～400mg/L；KH
2PO
4，100～250mg/L；
(2) micro-composition
FeSO
4·7H
2O，13.9～27.8mg/L；Na
2EDTA·2H
2O，18.65～37.3mg/L；KI，0～0.83mg/L；H
3BO
3，0～6.2mg/L；MnSO
4·4H
2O，11.2～25mg/L；ZnSO
4·7H
2O，4.3～10mg/L；Na
2MoO
4·2H
2O，0.25mg/L；CuSO
4·5H
2O，0.01～0.025mg/L；CoCl
2·6H
2O，0.025mg/L；
(3) organotrophy composition
Inositol, 500～1000mg/L; Nicotinic acid, 0.5～5mg/L; Pyridoxine hydrochloride, 0.5～1mg/L; Vitamin, 0.1～1mg/L; Glycine, 2.0mg/L;
(4) growth regulator
2,4 dichlorophenoxyacetic acid, 0.1～0.5mg/L; 6-benzyl aminopurine, 0.5～1.5mg/L; Dormin 0.01～0.3mg/L;
(5) carbon source: sucrose, 20～30g/L;
(6) peptizer: agar, 6～7g/L;
Final pH is adjusted to 5.6～5.8;
Prescription 2:
(1) macroelement composition
KNO
3，400～850mg/L；NH
4NO
3，150～350mg/L；CaCl
2·2H
2O，200～500mg/L；MgSO
4·7H
2O，150～400mg/L；KH
2PO
4，100～250mg/L；
(2) micro-composition
FeSO
4·7H
2O，13.9～27.8mg/L；Na
2EDTA·2H
2O，18.65～37.3mg/L；KI，0～0.83mg/L；H
3BO
3，0～6.2mg/L；MnSO
4·4H
2O，11.2～25mg/L；ZnSO
4·7H
2O，4.3～10mg/L；Na
2MoO
4·2H
2O，0.25mg/L；CuSO
4·5H
2O，0.01～0.025mg/L；CoCl
2·6H
2O，0.025mg/L；
(3) organotrophy composition
Inositol, 500～1000mg/L; Nicotinic acid, 0.5～5mg/L; Pyridoxine hydrochloride, 0.5～1mg/L; Vitamin, 0.1～1mg/L; Glycine, 2.0mg/L;
(4) growth regulator
α-Nai Yisuan 0.1～0.5mg/L, 6-benzyl aminopurine 0.5～1.5mg/L;
(5) carbon source: sucrose, 20～30g/L;
(6) peptizer: agar, 6～7g/L;
Final pH is adjusted to 5.6～5.8.
2. inducing culture as claimed in claim 1 is characterized in that: described macroelement composition is KNO
3, 600mg/L; NH
4NO
3, 200mg/L; CaCl
22H
2O, 220mg/L; MgSO
47H
2O, 370mg/L; KH
2PO
4, 170mg/L.
3. a peace ancestral spends tissue cultured seedling propagating method, comprises the steps:
(1) induce leaf or petiole to produce callus and indefinite bud
Utilize claim 1 or 2 described inducing cultures to induce and produce callus and indefinite bud: the potted plant seedling leaf that will nourish and generate or petiole are seeded on the substratum that adopts prescription 1 preparation, after cultivating for 3～8 weeks, induce the generation callus, or then quilt is induced the differentiation indefinite bud, though or generation callus, when not producing indefinite bud, then will be greater than 3～5mm
3Callus transfer on the substratum that adopt prescription 2 preparations, after cultivating through 6～8 weeks, callus is induced the differentiation indefinite bud again;
(2) adventitious shoot breeding
Above-mentioned callus with differentiation capability has promptly been differentiated the callus of indefinite bud, and subculture is cultivated on modified MS medium, propagation callus and breeding tissue cultured seedling fast; Callus is cultivated in 1: 4 ratio shoot proliferation after cultivating for 6～7 weeks on the modified MS medium; When being used for tissue cultured seedling and breeding fast, the callus subculture is in 1: 2 ratio subculture, and keeps the adventitious shoot less than 0.5cm length on callus;
Growth regulator composition in the described modified MS medium is: α-Nai Yisuan 0.1～0.5mg/L, 6-benzyl aminopurine 0.4～2.0mg/L; Carbon source: sucrose, 20～30g/L; Peptizer: agar, 4～6g/L; Macroelement composition, micro-composition, organotrophy composition are identical with corresponding composition in the MS minimum medium; Final pH is adjusted to 5.6～5.8;
(3) tissue cultured seedling root induction and transplanting
With length is that adventitious shoot more than 2～3cm is transferred in the root media, and after cultivating through 2～4 weeks, adventitious shoot produces new root, and new root growth to 0.5～when 1cm is long adopts the conventional tissue cultured seedling method for transplanting tissue cultured seedling of will taking root to be transplanted in the soil matrix;
Contain MS minimum medium composition, naphthylacetic acid 0.05～0.2mg/L, sucrose 10g/L, agar 7g/L that macroelement content reduces by half in the described root media, the final pH value regulates 5.6～5.8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100050177A CN100424169C (en) | 2005-01-31 | 2005-01-31 | Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100050177A CN100424169C (en) | 2005-01-31 | 2005-01-31 | Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1813525A CN1813525A (en) | 2006-08-09 |
| CN100424169C true CN100424169C (en) | 2008-10-08 |
Family
ID=36905978
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100050177A Expired - Fee Related CN100424169C (en) | 2005-01-31 | 2005-01-31 | Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100424169C (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823503B (en) * | 2012-09-26 | 2013-08-14 | 钦州市林业科学研究所 | Tissue culture medium for propagating anthurium buds by using buds |
| CN102893868B (en) * | 2012-10-18 | 2014-06-18 | 广东省农业科学院花卉研究所 | Method for embryogenic cells of anthurium scherzerianum to quickly proliferate and regenerate plant |
| CN103181326A (en) * | 2013-04-10 | 2013-07-03 | 苏州大学 | In vitro tissue cultivation method of potted anthurium andraeanum varieties |
| CN105010141A (en) * | 2015-07-09 | 2015-11-04 | 虞龙 | Anthurium rapid propagation method |
| CN107155552A (en) * | 2017-04-25 | 2017-09-15 | 马山县盛世农业发展有限责任公司 | Use the implantation methods of the elegant jessamine of tissue-cultured seedling |
| CN109906938B (en) * | 2019-03-07 | 2021-10-29 | 三明市农业科学研究院 | Anthurium germplasm resource in vitro preservation method |
| CN110063258A (en) * | 2019-05-23 | 2019-07-30 | 三明市农业科学研究院 | A kind of red fruit beadplant quick breeding method for tissue culture |
Citations (2)
| 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 |
| CN1552199A (en) * | 2003-12-19 | 2004-12-08 | 山西省农业科学院园艺研究所 | Quick reproduction of Anzu flower by cell embryo induction |
-
2005
- 2005-01-31 CN CNB2005100050177A patent/CN100424169C/en not_active Expired - Fee Related
Patent Citations (2)
| 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 |
| CN1552199A (en) * | 2003-12-19 | 2004-12-08 | 山西省农业科学院园艺研究所 | Quick reproduction of Anzu flower by cell embryo induction |
Non-Patent Citations (6)
| Title |
|---|
| 不同红掌品种的叶片、叶柄和茎段愈伤组织的诱导及植株再生. 兰芹英等.西北植物学报，第23卷第6期. 2003 |
| 不同红掌品种的叶片、叶柄和茎段愈伤组织的诱导及植株再生. 兰芹英等.西北植物学报，第23卷第6期. 2003 * |
| 安祖花叶片的离体培养. 高遐虹等.北京农学院学报，第10卷第2期. 1995 |
| 安祖花叶片的离体培养. 高遐虹等.北京农学院学报，第10卷第2期. 1995 * |
| 红掌叶片离体培养与植株再生研究. 吕复兵等.广东农业科学，第6期. 2002 |
| 红掌叶片离体培养与植株再生研究. 吕复兵等.广东农业科学，第6期. 2002 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1813525A (en) | 2006-08-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101587707B1 (en) | Producing method of orchid seedlings | |
| Boxus | The production of strawberry plants by in vitro micro-propagation | |
| CN101695283B (en) | Culture medium for tissue culture of pedicel buds of oncidium hybridum and tissue cultured seedling propagating method | |
| CN100424169C (en) | Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method | |
| CN103190347B (en) | Teapot dates tissue culturing method | |
| CN101983556A (en) | Spruce somatic embryogenesis and plant regeneration method | |
| CN105028193B (en) | A kind of utilization blueberry Lai Gexi blades induction produces the mating system of micro adventitious bud | |
| CN106665367B (en) | A kind of Golden Bell Tree quick breeding method for tissue culture | |
| JPS6137029A (en) | Regeneration of sunflower by germ and organ formation | |
| JP3330365B2 (en) | Culture fluid for Scutellaria plants | |
| Beruto et al. | In vitro culture of tree peony through axillary budding | |
| CN105379621A (en) | Efficient in-vitro plant regeneration method of adult high-quality single-plant Xiaoqiao oriental cherry of cerasus lannesiana var. speciosa | |
| CN1224314C (en) | Root inductive method for microbody reproduction of Japan dahurian larch | |
| CN101268758A (en) | Quick replication method for Xuan pawpaw tissue cultivation | |
| CN107155882A (en) | A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method | |
| CN114600772A (en) | Tissue culture method and rapid propagation method of michelia figo in remote mountains | |
| JP3706085B2 (en) | Medium for plants of the genus Camellia | |
| KR100457999B1 (en) | Propagation Method of Pinus rigida × P. taeda through Somatic Embryogenesis Technique | |
| HU183433B (en) | Process for producing multiplying material of digitalis lanata ehrh in tissue culture | |
| CN104255493B (en) | A kind of method of producing feather cockscomb thumb flower | |
| CN114467753B (en) | Tissue culture method of silvery deer maple | |
| CN110419447B (en) | Blueberry tissue culture method | |
| Economou et al. | Regeneration in vitro of oleaster (Elaeagnus angustifolia L.) from shoot tips of mature tree | |
| JP3715689B2 (en) | Mass propagation of Gmelina trees using tissue culture | |
| JPH0322931A (en) | Formation and proliferation of crown of plant of genus asparagus and proliferation of seedling |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: XIAO ZUNAN Effective date: 20130205 Owner name: BEIJING NORMAL UNIVERSITY Free format text: FORMER OWNER: XIAO ZUNAN Effective date: 20130205 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20130205 Address after: 100875 Beijing city Haidian District Xinjiekou Avenue No. 19 Beijing Normal University Patentee after: Beijing Normal University Patentee after: Xiao Zunan Address before: 100875 School of life sciences, Beijing Normal University, 19 Xinjie street, Haidian District, Beijing Patentee before: Xiao Zunan |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20081008 Termination date: 20210131 |