CN1341351A - Chinese sweetgum tissue culture and quick propagation method - Google Patents
Chinese sweetgum tissue culture and quick propagation method Download PDFInfo
- Publication number
- CN1341351A CN1341351A CN 01134054 CN01134054A CN1341351A CN 1341351 A CN1341351 A CN 1341351A CN 01134054 CN01134054 CN 01134054 CN 01134054 A CN01134054 A CN 01134054A CN 1341351 A CN1341351 A CN 1341351A
- Authority
- CN
- China
- Prior art keywords
- bud
- tissue
- medium
- hormone
- 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.)
- Granted
Links
Images
Abstract
The present invention is characterized by that on the basis of selecting dominant tree its respectively adopts the annual seedling stem apex, leaf blade and petiole, hypocotyl, male rhachis and immature embryo of Chinese sweetgum to make tissue cultivation. Said invention provides a seedling cultivating method for large-scal planting Chinese sweetgum, and its period is short, propagation coefficient is high and cost is low.
Description
The invention belongs to the forest tissue cultivating and seedling technology in the forest-science
Echinopodospora jamaicensis (Liquidambar formosana) is a kind of of Hamamelidaceae Liquidambar.The Liquidambar modern times have four main kinds, and they all are tall and big deciduous trees, are grown in semi-tropical warm area.Wherein satin walnut (Liquidambar styraciflua) is distributed widely in the high altitude mountainous area in south, North America, the southeast and Mexico, Central America to Honduras south; Storax (L.orientalis) is a kind of of little Ya Xiya, now is distributed in the osmanli west and south; What be distributed in China has echinopodospora jamaicensis (Liquidambarformosana) and two kinds of scarce calyx sweetgum (Liquidambar acalycina), and they are all very wide in the region that China distributes, to the east of Taiwan, to the border, Sichuan-Tibet, reach Guangdong and Hainan in the south, north reaches the Qinling Mountains, Basin of Huaihe River, and distribution is all arranged.
The tree-like grace of echinopodospora jamaicensis is dense, and the green and the end setting off each other after autumn, is one of fine tree species of building street tree, flower garden and amenity forest.Its trunk is perfectly straight satisfactory, and the grain of wood is careful, though easily buckling deformation can be preserved for a long time in aeration-drying place, is the last handy material of building, and the title of " the last maple of Wan Niange " is arranged.Sweetgum is a first-class material of making plywood, also is the ideal material of manufacturing large-scale extranal packing boxes such as tealeaves, food simultaneously.Brush woods such as branch, the tip can be cultivated mushroom, auricularia auriculajudae; The standing tree of living can be tapped maple fat, for system spices; Root, skin, fruit can be used as medicine, and fruit medicine name " plays both sides of the street ", and the effect of dispelling wind and removing obstruction in the meridians, the logical breast of Li Shui is arranged.The tree root of sweetgum bury very dark and well developed root system can drought-resistant, barren and farmland running to weeds, and its adaptability is similar to masson pine with vitality.Property happiness sunlight, also ability shade, wind resistance, cold-resistant.Fertile, moistening neutrality and acid ground are born in happiness, but also can grow on barren grit soil or heavy earth.Rudiment power and natural regeneration ability are strong, on cutting blank, open forest land and deserted mountain natural regeneration good, become pioneer tree species or dominant tree species.Sweetgum is in natural forest, and the height of tree and breast diameter growth 10 years are slower before death, and 10～50 years is that speed is given birth to the stage when living, and 20～30 years be fast life peak period when livings.Under suitable land occupation condition, can become a useful person about 20 years.
The echinopodospora jamaicensis monoecism.General employing seminal propagation.Observation according to recent years finds that the seed production of echinopodospora jamaicensis has the branch of biennial bearing, and biennial bearing cycle basic fixed (2 years).In seed good year, on the branch at each position of bearing tree tree crown aggregate fruit is arranged all; In seed off year, have only the minority aggregate fruit on the branch at the top of tree crown give birth to.The solid characteristic of echinopodospora jamaicensis and seed orchard breeding production research still are in the incipient stage at present, and supply falls short of demand for breeding.Therefore, on the basis that the echinopodospora jamaicensis superior families is selected, be necessary to carry out the vegetative propagation research and the technology development work of echinopodospora jamaicensis.
Tissue culture technique is one of main contents of present forestry biotechnology, and its main application is exactly large-scale breeding and the genetic transformation of forest.1902, under the promotion of the cell theory that Scheleiden and Schwann grew up, German plant physiologist Haberlandet proposed the organ of higher plant and has organized and can constantly cut apart until the viewpoint of assigning to individual cells.Because selected experiment material was that the cell and the medium that have highly broken up are too simple at that time, did not comprise and induced mature cell to divide necessary somatotropin, did not observe cell division in experiment.Up to 1934, White cultivated the stripped tip of a root success of tomato.
Twentieth century the mid-1930s is to the latter stage fifties, by broad research to condition of culture and medium composition, particularly to vitamin B group, the research that the growth hormone and the basic element of cell division act in tissue culture, realized control to cells in-vitro growth and differentiation, thereby tentatively established the technical system of tissue culture, for later development is laid a good foundation.The forties and the initial stage fifties, the researcher who is active in the Plant Tissue Breeding field is representative with Skoog, and the purine substance that to the effect that utilizes of research is handled tobacco marrow callus with the growth of control tissue and the formation of bud.Skoog (1949) and Skoog are with discovery such as Cui Cheng (1951), adenine and adenosine not only can promote callus Growth, and can also remove growth hormone heteroauxin (indole-3-acetic acid in the medium, hereinafter to be referred as IAA) inhibitory action that bud is formed, the formation of induced bud, thus the ratio of determining adenine and growth hormone is one of essential condition of control bud and root formation.These experimental results have caused the discovery of later kinetin and have utilized kinetin later on and growth hormone breaks up carrying out of work tissue culture middle controller official.In the forties, another development of plant tissue culture technique is that Overbeek etc. (1941) joins coconut milk in the medium first, make the heart-shaped phase rataria of datura can cultured in vitro to ripe.To the beginning of the fifties,, thereby make coconut milk all obtain extensive use in the every field of tissue culture because Steward etc. have also used this material in the carrot tissue culture.Nineteen fifty-two, Morel and Martin confirm first, by the cultured in vitro of shoot apical meristem, and can be by obtaining nontoxic plant in the dahlia that is subjected to virus infection.1953～1954 years, Muir carries out unicellular cultivation and obtains initial success, method is that the callus of Aztec marigold and tobacco is transferred on the liquid nutrient medium, be placed on the shaking table and vibrate, make historrhexis, the cell suspending liquid that formation is made up of unicellular and cell aggregation is bred by successive transfer culture then.Nineteen fifty-five, Miller etc. are by isolating a kind of known first basic element of cell division in the herring sperm dna, and it is named are kinetin.Now, have with Kinetins existing multiplely like active synthetic or native compound, they are generically and collectively referred to as the basic element of cell division (cytokinin).Use this class material, can induce cell ripe and well differentiated tissue to divide.Nineteen fifty-seven, Skoog has proposed the notion that relevant plant hormone control organ forms with Miller, point out in the tobacco myeloid tissue is cultivated, the differentiation of root and stem is the function of growth hormone pair cell mitogen ratio, regulate the differentiation that the relative concentration of material can be controlled organ by changing this two class in the medium, hestening rooting when this ratio is high promotes the differentiation of bud when low, when the two concentration equates, organize and then tend to grow in a kind of structureless mode.Proved afterwards, hormone is adjustable, and organogenetic notion is all applicable for most species, be owing to the interior unboiled water of these hormones in different tissues is flat different, thereby concerning a certain concrete form generating process, the level of their desired exogenous hormones also can be different.1958～nineteen fifty-nine, Reinert and Steward report respectively, have formed somatic embryo in carrot callus cultivates, and this is a kind of regeneration that forms plant by the differentiation of bud and root that is different from.
Twentieth century is since the sixties, and tissue culture technique has obtained developing rapidly aspect following three:
1. the protoplast cultivation obtains important breakthrough---and nineteen sixty, people such as Cocking utilize fungal cellulase separating plant protoplast to succeed.1971, Takebe etc. have obtained whole plant by protoplast first on tobacco, this has proved in theory that not only the protoplast of no wall has totipotency equally except that somatic cell and reproductive cell, and can provide desirable acceptor material for the importing of foreign gene in practice.After 1985, cultivate in succession as the protoplast of the cereal plants of grain and feed main source and to succeed; Aspect woody plant, mainly also obtaining success aspect the protoplast cultivation of some deciduous species.The success that protoplast is cultivated has promoted the development of somatic cell integration technology.1972, Carlson etc. obtained first burdo by the fusion of protoplast between two tobacco species.
2. anther culture achieves notable results---and 1964, Guha and Maheshwari report became embryo by the anther culture that exsomatizes by microspore direct development in the golden flower of Nan Yang.1967, Bougin and Nitsch obtained complete tobacco plant by anther culture.Because the important function of monoploid in sudden change selection and isozygotying of acceleration heterozygote process, the research in this field obtained developing rapidly in the whole seventies.
3. little propagating technology is used widely---and nineteen sixty, Morel has proposed the method for an isolated vegetative propagation orchid, and reproduction coefficient is high.Because this method has great practical value, adopted very soon by orchid production person, set up orchid industry rapidly.At present, except that orchid, aspect other many ornamental plantss and economic crops and forest (willow, eucalyptus), little propagating technology has also reached the production scale of batch production.
Organ generation regeneration plant approach in the Plant Tissue Breeding needs separated bud induction period of experience and root induction phase.Organ has indirect and direct dual mode.Under indirect mode, indefinite bud induces from callus.Under direct mode, the bud of regeneration is to induce from the bud that has existed.When bud is from those positions (as needle, cotyledon) that should not produce bud when inducing, this bud is called indefinite bud, and when bud be when regenerate in the position that can produce bud usually, this bud is called lateral bud and axillalry bud.
From callus surface or inner bud and the root of forming, the ratio of the basic element of cell division and growth hormone in the medium is depended in its differentiation.In the experiment of classics, Skoog and Miller utilize the tobacco marrow callus of cultured in vitro, usually study these two kinds of exogenous hormones to the organogenetic control of leaf explant by applying variable concentrations combined I AA and excitement.The result shows that high-caliber kinetin (1.0mg/l) can promote to form bud.When having IAA to exist, the kinetin of medium level (0.2mg/l) helps the propagation of callus, the generation of root is then induced in the independent effect of IAA, this experiment confirm the regulating and controlling effect of interaction partners tobacco healing tissue differentiation of the basic element of cell division and growth hormone.But different Plant Tissue Breeding systems have than big-difference the desired level of these two kinds of exogenous hormones, and obviously this is relevant with the level of endogenous hormones in the tissue.Concerning most vegetable materials, the isolated organ differentiation needs the effect of certain external source growth regulatory substance usually.
In twentieth century sixties, Torry proposes a hypothesis, thinks that isolated organ arises from a kind of meristematic tissue group that forms among callus, or is called the class meristematic tissue.These meristematic cell groups can produce reaction and form original hase different stimulated.Finally being divided into root as for these original hases still is bud (perhaps embryoid), then depends on the acting in conjunction of internal factor and outside stimulus.Later many histological observations that organ in the tissue culture is formed show before the growth of adventive root or indefinite bud, the appearance in local meristematic tissue zone is arranged all in the callus.Understand not to the utmost for the definite reason and the condition that form this kind meristem zone territory and site at present, but appear at the fact of cultured tissue and medium intersection usually according to these meristematic tissue, the someone infers the diffusion and the physiological gradient that forms may play effect in the position that decision class meristematic tissue forms in tissue of some active substance in the medium.
In most of dicotyledons, when growth hormone in the medium/when basic element of cell division ratio was 1～100, in vitro tissue often was in callus induction and vegetative state.Otherwise, when the basic element of cell division/when the growth hormone ratio reaches 10～100, then promote the differentiation and the formation of indefinite bud.Therefore concerning these plants if just callus is transferred to from the medium of high growth hormone on the differential medium of the high basic element of cell division can induced bud generation.But under situation about having, then need in medium, economize and omit a kind of exogenous hormone and just can reach this purpose.But this exogenous hormone condition of the bud differentiation of exsomatizing of inducing should not regarded blanket rule as.Because in reality is cultivated, many exceptions that do not meet these rules are arranged.Street thinks, by regulating exogenous auxin/basic element of cell division ratio, perhaps have following several by the reason of removing the differentiation that growth hormone still can not induced bud: accumulated more endogenous hormones in (1) callus, the exogenous hormone ratio that is adopted is not enough to offset their startups to atomization; (2) at the additional other exogenous hormone of a certain specific material require; (3) physical condition of a certain trophic factor or cultivation is unfavorable for the startup of atomization.
Kinetin is the most frequently used organogenetic exogenous hormone of adjusting.Other compounds with induced bud differentiation also have benzyladenine (benzylaminopurine is hereinafter to be referred as BA), 2-isopentennyladenine (2-ip) and zeatin (zeatin is hereinafter to be referred as ZT) etc.In addition, some phenylurea compounds are as the be formed with good effect of thiadiazole phenylurea (thidiazuron is hereinafter to be referred as TDZ) to lateral bud differentiation in the organ generation and indefinite bud.In addition, endogenous ethylene also can influence the bud differentiation.Found in Plant Tissue Breeding in early days, this liquid hormone substance can check organ and take place.But form the stage at original hase, ethene then has the effect that promotes growth.
The metabolism of carbohydrate also is counted as the factor that bud forms that influences.The external source carbohydrate is also being exercised the function of regulating osmotic pressure except as the energy.In the tissue of cultured in vitro,, thereby influence its growth and form even the change of slight osmotic pressure also can cause the variation of callus biochemical metabolism.
The formation of root is the organ occurring mode that the frequency of occurrences is the highest in the Plant Tissue Breeding.Two kinds of different root ex vivo differentiation forms are arranged.A kind of is thereby that the bud that has formed bears root formation regrowth in cultivation, is called take root (rooting).Another kind of then be to be divided into the original hase of root and further to grow by the class meristematic tissue in the callus to form root, be called root generation (rhizogenesis).By tissue culture elder generation evoking adventive bud, and then dissolving root in its base section, is a kind of common isolated organ occurring mode.At this moment, the differentiation of bud and growth obviously can change the composition and the ratio of endogenous hormones in the tissue, thereby the formation to root exerts an influence, and this process that obtains regeneration plant through taking root by indefinite bud also is to utilize tissue culture to carry out one of important channel of clonal reproduction.Root generation phenomenon in the callus is often not as the general and easy acquisition like that of taking root, and the differentiation capability of callus root can weaken gradually along with the increase of subculture number so that forfeiture.
In the forest tissue-culturing quick-propagation, producing regeneration plant by the organ occurring mode is the most successful cultured in vitro modes of reproduction.Coniferous tree is as main reproducting tree species, and its group culturation rapid propagating technology begins starting in the 60 to 70's of twentieth century, and is rapidly developed.1958, the Brown of georgia ,u.s.a university and Lawrence tested out the BL medium that is applicable to the pine tree tissue culture.In 1975, Sommer etc. realized obtaining regeneration plant by the mode of adventitious shoot regeneration first in pine tree.Owing to acerose fertility can descend along with the increase of the age of tree, once adopted juvenile form material (comprising the segment of the seedling that mature zygotic embryos and germination seed generate) so be most appropriate to the material of the quick breeding of coniferous tree always.The method of this quick breeding is suitable for the expanding propagation of select tree seed most.Amerson had once reported and has adopted this technology to come the method for quick propagating torch pine.The torch pine seed was sprouted under aseptic condition after 5 days, downcut cotyledon as explant.Cotyledon is cultivated on the additional improvement BL medium that 44 μ M are arranged and is induced meristematic tissue, after 14 days, it is transferred to does not contain BA but adds on the medium of active carbon (active carbon is used for eliminating the effect of BA).On this bud differential medium, can observe from meristematic tissue punishment and dissolve indefinite bud, and begin elongation.The result shows that after through 4 months cultivation, on average each embryo can form 46 buds, 48% bud wherein, and its length is more than 0.5cm.When 1～2cm is grown in blastogenesis, be used for root induction.In the field contrast experiment who is done with the seedling and the same family half sibs seedling from seed of group training generation, the seedling that the training of discovery group produces is grown in first season of growth and is obviously lagged behind, but its growth significantly increases than seedling from seed in ensuing 2～4 years.As a result, in 4～6 years, both total growth recruitments are similar substantially.Lag behind the seedling of seed source although this seedling that shows that the group training is originated is grown in early days, when reaching rotation, the difference of the two is little.The seedling of in the contrast experiment, also observing group training source in addition growth regulation in the time of 2 years the seedling than seed source show more ripe feature.
Be easier to relatively although be used for quick breeding as explant, undertake a certain risk for it is worth without its heredity of check with the juvenile form material.In this case, adopt that tissue culture mode is fast numerous just to become more valuable through the bearing tree of field inspection for many years.But because the forest provenance, the difference of the growth conditions and the age of tree when carrying out fast numerous experiment between different forests, is difficult to find a kind of blanket culture technique.At present, fast numerous system of the adult select tree of Chinese larch (Sequoia sempervirens), maritime pine (P.maritima) and pine (P.radiata) sets up, and its corresponding field trial is also launched (Dunstan D.J., 1988).Gupta and Duzan had once introduced the method for the adult select tree of a kind of in-vitro propagate pesudotsuga taxifolia.At first get the stem section of the long band joint of 2～3cm from the big tree that gives birth to the sixties, cultivate on the DCR of no hormone medium the sterilization back.After 3～4 weeks, the bud that newly grows is transferred on the medium of no BA additional activity charcoal, impels bud to continue elongation.Shen Chang bud is transferred to the generation that promotes axillalry bud on the medium of 1 μ MBA then.The stem section of getting new generation simultaneously is used for inducing of bud.The bud that is used to take root is about 0.3～0.4cm is long, but rooting rate is very low.Have only 20% bud to take root after 12 weeks.
21st century the fifties end, Steward etc. (1958) almost find from the carrot root cells of cultured in vitro to produce a kind of structure similar to embryo with (1959) such as Reinert simultaneously, and observe by this structure and grow up to complete plant.Generally this structure is called somatic embryo or embryoid (Street andWithers, 1974 now; Reinert, 1978; Zhu Cheng, 1978).Embryoid can be divided into two classes by its source: a class is by the dliploid somatic cell generation of the various organs of common plant spore body, is commonly referred to somatic embryo, can develop into normal plant thus; Another kind of is the embryoid that is produced by haploid cells such as microspore or its cleavage products, abbreviates pollen embryo usually as, can develop into haplobiont, could normally blossom and bear fruit but generally must carry out chromosome doubling.Also in some plants, also obtain embryoid in addition, but develop into plant by embryoid with failing from the endosperm cultivation.Zhu Cheng (1978) thinks that the definition of embryoid comprises 3 implications: (1) embryoid is the product of tissue culture, is only limited in the tissue culture scope and uses, and is different from apomictic embryo; (2) embryoid originates from non-zygote cell, is different from zygotic embryo; (3) process of the formation of embryoid experience embryonic development is different from the sprout that breaks up in the tissue culture.
The body embryo takes place owing to can obtain more nutrition propagule in the shorter time, and the bigger genetic gain of acquisition, and because the body embryogenesis culture also is the important source of separating protoplast, can be used for genetic transformation and the somatic hybridization research of forest, therefore utilize forest body embryo to carry out a kind of important means that the forest vegetative propagation is following Developing Clonal Forestry.Research takes place and starts from the seventies later stage in the body embryo of forest, but is just developed rapidly up to back several years of the eighties, and the acquisition howling success.In the coniferous species, since reported first such as Hakman in 1985 induced body embryo and regeneration of plantlet by Norway spruce (Picea abies) immature zygotic embryos, research took place and has obtained the progress that attracts people's attention in acerose body embryo.According to the preliminary statistics, from abies (Abies), Larch (Larix), Picea (Picea), Pinus (Pinus), Pseudotsuga (Pseudotsuga) has produced somatic embryo with at least 20 kinds of different coniferous tree explants cultivations that sequoia sempervirens belongs to (Sequoia).Yang Jinling etc. (1997) cultivate through the body embryo plant that regenerates with Picea meyeri (Picea meyeri) mature zygotic embryos.It is that the explant cultivation obtains the body embryo that Rajbhandari (1997), Guevin (1997) utilize Abies fraseri (Abies frasi) unmature subleaf zygotic embryo in early stage and mature zygotic embryos respectively.In deciduous species, Castanea (Querus), Populus (Populus), eucalyptus belongs to (Eucalyptus), Aesculus (Aesculus), genus hevea (Hevea), santal belongs to (Santalum), Liquidambar (Liquidambar), Liriodendron (Liriodendron), Corylus (Corylus) etc. is observed the body embryo and is taken place or the plant that regenerates in tissue culture.
Since Hakman in 1985 reports that at first usefulness Norway spruce (Picea abies) immature zygotic embryos is as explant induction body embryo, immature embryo, mature embryo, seedling hypocotyl and the juvenile form materials such as earsh organ or regrowth organ of adopting as explant more in forest body embryo is studied, thereby reduced the method that from immature seed, strips this labor intensive of rataria, and can be provided for the explant (Tautorus, 1991) that the body embryo takes place throughout the year.Because the juvenile form material is without field inspection, heredity is worth unknown, so will be applied to production practices from its body embryo that induces, bear great risk, might cause immeasurable loss.In this case, for avoiding loss, general way is: at first induce the embryo culture from the juvenile form material, after enlarging propagation, each genotypic embryo culture keeps a duplicate samples low temperature and preserves (liquid nitrogen,-196 ℃), induce the body embryo from remaining embryo culture then, body embryonic development maturation germination Cheng Miaohou is used for afforestation.Through the field trial in 5～15 years, pick out the good genotype of minority according to the data that clonal test obtains, utilize these excellent genes types inductor embryo after the embryo culture that low temperature is preserved is bred in a large number to be applied to produce (Gupta, 1993) again.Though avoided loss like this, slowed down the progress of genetic improvement.Forest body embryo is applied to production practices, is ideal material from the explant of bearing tree.Because bearing tree has passed through the check in field for many years, the resistance of the output of product, quality and tree body is all known, can select best individuality to breed, to guarantee the economic benefit after body embryo seedling is afforested.But difficulty especially takes place again in the embryo of bearing tree explant, and Chen Zhenghua (1990) proposes bearing tree explant recovery embryo three ways: (1) utilizes the rejuvenation system of bearing tree self.As obtaining somatic embryo easily as explant with premeiotic immature inflorescence or postmeiotic megarchidium tissue; (2) with keeping the part of juvenile form in the bearing tree as explant.Often keeping juvenile form at the tissue at the low position of tree body such as sleeping bud and root turion, their embryo's generating ability is forfeiture not, therefore gets the bearing tree sprout tillers and also might make the success of body embryonal induction as explant; (3) can obtain embryo callus from the bearing tree tissue through the long-term subculture cultivation.
The woody plant that the part that table 1 has been reported utilizes bearing tree explant induction body embryo to take place
Botanical name explant author scruboak (Quercus ilex) blade Feraud, 1989 Norway spruces (Picea abies) needle, bud Wescott, 1994 white poplars * canine tooth poplar blade Michler, 1991 (Populus alba x P.grandkenta) Lombardy poplar * distant poplar blade Park, 1989 (Populus nigra x P.maximowiczii) olive (Olea europaea) petiole Rugini, 1995 Hevea rubbers (Hevea brasiliensis) secundine Michayx, 1995 wormwood artemisia willows (Salix viminalis) gynoecium Gronroos, 1989 European horse-chestnuts (Aesculus hippocustamum) filigree Jorgensen, 1989 cocoas (Theobroma caca) petal, staminodium, filigree Lopez-Baez, 1993 oil palms (Elaeis guineensis) prematurity female inflorescence Teixieira, the male catkin Gingas of 1994 swamp white oaks (Quercus bicolor), the male catkin Gingas of 1991 American Red oaks (Q.rubra), the male catkin Gingas of 1991 American Black oaks (Q.velutina), 1991 satin walnuts (Liquidambar styraciflua) prematurity male inflorescence Merkle, the old positive China of 1998 rubber (Hevea brasiliensis) flower pesticide, Liu Guizhen etc., 1989 teaks (Tectona grandis) terminal bud Rijuta, 1996
In the at present relevant all kinds of documents and materials of plant tissue culture technique, except that there was the relevant patented technology report of organizing training on satin walnut (Liquidambar styracifua) in the U.S., Shang Weijian organized the complete set technology of training and delivers on echinopodospora jamaicensis (Liquidambar formosana).
Main purpose of the present invention is to carry out the multiple target breeding research of echinopodospora jamaicensis (Liquidambr formosana), on the basis that superior families is selected, carry out the exploitation of echinopodospora jamaicensis tissue culture and little propagating technology, seek the packaged technology that is applicable to the echinopodospora jamaicensis tissue-culturing quick-propagation.
Major technique of the present invention is characterised in that, on the basis that select tree is selected, adopts 1 year living seedling stem apex of echinopodospora jamaicensis respectively, blade and petiole, and hypocotyl, male inflorescence axle and immature embryo carry out tissue culture.
1. adopt 1 year living seedling stem apex of echinopodospora jamaicensis to carry out tissue culture.Stem apex growth and the used minimal medium of bud differentiation and proliferation are WPM (Woody Plant Medium, be the woody plant tissure medium) and MS medium (Murashing and Skoog medium, hereinafter to be referred as MS), additional plant hormone is that concentration is the BA of 0～2.0mg/l, the naa of 0～0.2mg/l (naphthylene acetic acid, hereinafter to be referred as NAA), remove the plant hormone combination that BA concentration is lower than NAA.In addition, the TDZ of the additional 0.01～0.1mg/l of WPM medium.
2. the blade and the petiole that adopt 1 year living seedling stem-tip tissue of echinopodospora jamaicensis to turn out carry out tissue culture.It is the WPM medium that excised leaf, petiole organ take place to cultivate used minimal medium, and additional plant hormone combination is the NAA of 0～0.1mg/l, the BA of 0.5～5.0mg/l.In addition, add the TDZ of 0.01mg/l separately.
3. take off plumular axis after adopting the echinopodospora jamaicensis seed under aseptic condition, to sprout, be seeded in the method for carrying out tissue culture on the callus inducing medium.The minimal medium that is adopted is MS and MS/2.
4. adopt when mixed bud has just begun to expand outside male inflorescence stretches out mixed bud, get the male inflorescence axle and be used for the method that callus induction carries out tissue culture.The minimal medium that is adopted is MS/2.
5. adopt at seed after 4～6 weeks of pollination, when the cotyledon type embryo to seed, occurring, take out rataria and be used for the method that callus induction carries out tissue culture.The minimal medium that is adopted is MS/2.
The hormone that callus induction is used is the 2.4-D of 0.5～3.0mg/l, the BA of 0～2.0mg/l.The hormone that adventitious bud inducing is used is the ZT of 0.5～1.0mg/l, the BA of 0.5～1.0mg/l and the NAA of 0.1mg/l.
WPM (Woody Plant Medium) is that the standard recipe of woody plant tissure medium is as follows:
NH
4NO
3 400mg/l
Ca(NO
3)
2·4H
2O 556mg/l
CaCl
2·2H
2O 96mg/l
MgSO
4·7H
2O 370mg/l
KH
2PO
4 170mg/l
K
2SO
4 990mg/l
Na
2-EDTA 37.3mg/l
FeSO
4·7H
2O 27.8mg/l
MgSO
4·H
2O 22.3mg/l
ZnSO
4·7H
2O 8.6mg/l
H
3BO
3 6.2mg/l
Na
2MoO
4·2H
2O 0.25mg/l
CuSO
4·5H
2O 0.25mg/l
Inositol 100mg/l
Nicotinic acid 0.5mg/l
Thiamine hydrochloride 1mg/l
The standard recipe of glycine 2mg/l MS medium (Murashing and Skoog medium) is as follows:
NH
4NO
3 1650mg/l
KNO
3 1900mg/l
CaCl
2·2H
2O 440mg/l
MgSO
4·7H
2O 370mg/l
KH
2PO
4 170mg/l
KI 0.83mg/l
H
3BO
3 6.2mg/l
MnSO
4·H
2O 16.9mg/l
ZnSO
4·7H
2O 8.6mg/l
Na
2MoO
4·2H
2O 0.25mg/l
CuSO
4·5H
2O 0.025mg/l
CoCl
2·6H
2O 0.025mg/l
FeSO
4·7H
2O 27.8mg/l
Na
2-EDTA 37.3mg/l
Inositol 100mg/l
Nicotinic acid 0.5mg/l
Thiamine hydrochloride 0.1mg/l
Puridoxine hydrochloride 0.5mg/l
Glycine 2mg/l MS/2, promptly the standard recipe of the MS medium (Murashing and Skoog medium) that reduces by half of macroelement is as follows:
NH
4NO
3 825mg/l
KNO
3 950mg/l
CaCl
2·2H
2O 220mg/l
MgSO
4·7H
2O 185mg/l
KH
2PO
4 85mg/l
KI 0.83mg/l
H
3BO
3 6.2mg/l
MnSO
4·H
2O 16.9mg/l
ZnSO
4·7H
2O 8.6mg/l
Na
2MoO
4·2H
2O 0.25mg/l
CuSO
4·5H
2O 0.025mg/l
CoCl
2·6H
2O 0.025mg/l
FeSO
4·7H
2O 27.8mg/l
Na
2-EDTA 37.3?mg/l
Inositol 100mg/l
Nicotinic acid 0.5 mg/l
Thiamine hydrochloride 0.1mg/l
Puridoxine hydrochloride 0.5mg/l
Glycine 2mg/l
Below be the specific embodiment that adopts the technology of the present invention:
Embodiment 1: adopt 1 year living seedling stem apex of echinopodospora jamaicensis to carry out tissue culture
At late March, 1 year living seedling plant is chosen the branch of active growth from the echinopodospora jamaicensis field of robust growth, gets its 5cm top, through 75% ethanol 30s, after 0.1% mercuric chloride, the 2 min sterilization, aseptic water washing 4～5 times, the stem apex that cuts 1.0～1.5cm is as explant.
Stem apex growth and the used minimal medium of bud differentiation and proliferation are WPM and MS medium, additional plant hormone combination is respectively BA0,0.1,0.5,1.0,2.0mg/l, NAA 0,0.05,0.2mg/l, removal BA concentration is lower than the plant hormone combination of NAA, WPM cultivates additional TDZ 0.1 in addition, and 0.01mg/l contains sucrose 3% in the medium, agar 0.65%, pH transfers to pH5.8 before the sterilization, 121 ℃ of high temperature, autoclaving 16min, 24～26 ℃ of cultivation temperature, illumination 16hr/d is about illuminance 2000lx.
If material after several days brown stain takes place in inoculation, material in time can be transferred on the fresh culture.Cultivate the differentiation bud test-tube plantlet that obtained echinopodospora jamaicensis in about 15 days afterwards.
Its different hormone concentrations combination to the influence of Shoot Tip Culture bud quantity of differentiation referring to accompanying drawing 1;
Its different hormone concentration combinations are broken up the influence of height referring to accompanying drawing 2 to the Shoot Tip Culture bud.
Higher (1.0, on medium 2.0mg/l), it is normal bud when just differentiation has been come out that the part bud is arranged, and is vitrifying bud transparent, many water but gradate as time passes in BA concentration.By reducing BA concentration, can effectively prevent the generation of vitrifying bud to below the 0.5mg/l and increase the content to 7% of agar in the subculture medium.
Table 2 different B A concentration is to the vitrified influence of bud
The BA stem apex is counted bud differentiation number vitrifying bud vitrifying rate (mg/l) (individual) (individual) (individual) (%)
0 12 12 0 0
0.1 12 15 2 13.3
0.5 12 43 9 20.9
1.0 12 60 37 61.7
2.0 12 49 33 67.4
Embodiment 2: the blade and the petiole that adopt 1 year living seedling stem-tip tissue of echinopodospora jamaicensis to turn out carry out tissue culture
On the echinopodospora jamaicensis differentiation bud that goes out by Shoot Tip Culture, choose the tender leaf that launches or just launched, the band petiole downcuts together.Under aseptic condition, with scalpel blade and petiole are separated, stay 1～2mm petiole end on the blade, then blade laterally is cut into the wide slice of 3～5mm with scalpel, adaxial and its surface lies against on the organ generation medium up, and the segment that petiole is cut into about 5mm lies against on the organ generation medium.
Used minimal medium is WPM, and additional plant hormone combination is NAA 0,0.1mg/l, BA0.5,1.0,2.5,5.0mg/l.In addition, additional separately TDZ 0.01mg/l.Contain sucrose 3% in the medium, agar 0.65%, medium sterilization and condition of culture are with embodiment 1.
Cultivated for 6 weeks at blade, leafstalk culture can obtain the test-tube plantlet of echinopodospora jamaicensis after 8 weeks.
Its different hormone concentrations combination to the influence of excised leaf, petiole differentiation referring to accompanying drawing 3
As can be seen from Figure 3, be treated to optimum with BA2.5mg/l and NAA0.1mg/l in the reason throughout.Under this condition, on average each blade can be induced 6.0 indefinite buds, and on average each petiole can be induced 4.2 indefinite buds, and the organ generating ability of blade is than being eager to excel of petiole, and the regeneration frequency of indefinite bud all was that blade is higher than petiole during all were handled.
Embodiment 3: cultivate the echinopodospora jamaicensis test-tube plantlet with the seed hypocotyl
Behind echinopodospora jamaicensis seed flowing water flushing 3hr, through 75% ethanol 30s, after 0.1% mercuric chloride, the 10 min sterilization, aseptic water washing 4～5 times is seeded on the MS medium of no hormone.After 1 week, seed begins to sprout, and when seedling to be sprouted grows to 1～2cm, takes off plumular axis and is cut into 3～5mm segment as explant.The minimal medium that callus induction is used is MS and MS/2.Used hormone is 2 of 0.5～3.0mg/l in the medium, 4-D, the BA of 0～2.0mg/l.The hormone that adventitious bud inducing is used is the ZT of 0.5～1.0mg/l, the BA of 0.5～1.0mg/l and the NAA of 0.1mg/l.
Callus has adopted three kinds of pretreatment modes when going on the differentiation adventitious buds medium by the callus proliferated culture medium:
1. do not have preliminary treatment and directly shift, this mode in contrast;
2. transfer to earlier on the no hormone culture-medium and cultivated 5 days;
3. transfer to earlier on the medium of no hormone and additional 3% active carbon and cultivated 5 days.
Three kinds of pretreatment modes to the influence of callus differentiation adventitious buds referring to accompanying drawing 4.
Embodiment 4: cultivate the echinopodospora jamaicensis test-tube plantlet with rhachis
Early March is adopted down the echinopodospora jamaicensis spray in indoor water planting, gets mixed bud weekly 1 time, and the male inflorescence axle of getting wherein is used for experiment.When mixed bud did not launch, whole mixed bud sterilization stripped out the male inflorescence axle then and is inoculated on the MS/2 medium under aseptic condition, and the mixed bud perula is launched, and after male inflorescence stretches out, directly cuts the male inflorescence axle, was cut into the long segment of 3-5mm after the sterilization as explant.
The minimal medium that callus induction is used is MS/2.Used hormone is 2 of 0.5～3.0mg/l in the medium, 4-D, the BA of 0～2.0mg/l.The hormone that adventitious bud inducing is used is the ZT of 0.5～1.0mg/l, the BA of 0.5～1.0mg/l and the NAA of 0.1mg/l.
Embodiment 5: cultivate the echinopodospora jamaicensis test-tube plantlet with immature embryo
Gather aggregate fruit since the first tenday period of a month in May (pollination 4～6 weeks of back), adopt weekly 1 time, to seed, have cotyledonary embryos to occur.Be loaded in the plastic sack after the aggregate fruit that collects wraps up with wet gauze and refrigerated for 1 weeks, behind the surface sterilizing, take out seed at 4 ℃, peel off kind of a skin, or strip out seed, behind the surface sterilizing, take out embryo and insert evoked callus in the MS/2 medium that adds hormone as explant.
Used hormone is 2 of 0.5～3.0mg/l in the medium, 4-D, the BA of 0～2.0mg/l.The hormone that adventitious bud inducing is used is the ZT of 0.5～1.0mg/l, the BA of 0.5～1.0mg/l and the NAA of 0.1mg/l.
Embodiment 6: the taking root and transplant of test-tube plantlet
When the blastogenesis on the test-tube plantlet is grown to 2～3 centimetres, choose the bud that growth is vigorous, the children is tender, scale off, carry out root induction.Root media is WPM medium and the additional indolebutyric acid (indole-3-butyric acid is hereinafter to be referred as IBA) 0,0.75,1.0 that macroelement reduces by half, 2.0mg/l.Behind the seedling rooting, move to vermiculite/perlite (1: 1, in matrix V/V), keep humidity.Can transplant plantation according to a conventional method after taking root.
After test-tube plantlet is inoculated into and carries out root induction on the root media that adds variable concentrations IBA, seedling base portion elder generation adularescent adventive root projection (processing that IBA concentration is high has callusization to a certain degree), the seedling base portion bears many radial white roots then.
Table 2 IBA to the effect IBA rootage duration of rooting of vitro seedling take root the mean elements seedling base portion on the every offspring of percentage callus degree (mg/l) (d) (%) (bar) 0 12 37.3 3.8 do not have 0.75 14 96.1 5.3 do not have 1.0 18 85.2 3.7 slight 2.0 20 81.8 2.1 serious
The result shows that the echinopodospora jamaicensis rooting of vitro seedling is not very strict to the requirement of IBA concentration.IBA during by 0～2.0mg/l rooting rate all more than 37.3%.The Huang phenomenon of withering takes place going to root media in the seedling of not taking root in experiment in the time of 5～10 days, yellow leaf comes off, the base portion blackout.Experiment is found, the state that grows of the seedling that selection is used to take root and the Huang phenomenon of withering is in close relations, select growth vigorous, blade is peak green and the long young tender attitude test-tube plantlet root induction of internode, can obtain higher rooting rate, and the death of withering of the short stereotype test-tube plantlet most Huangs in the root induction process of leaf dark green, internode has only indivedual rooting of vitro seedling.
Also can cut eugonic 2～3 centimetres bud directly is inserted into vermiculite/perlite of the bacterium of having gone out (1: 1, in matrix V/V), keeping temperature was that 20～30 ℃, matrix keep moistening.Can take root after about 10 days.Can transplant plantation according to a conventional method after taking root.
The WPM medium is good than the MS medium for the effect of Shoot Tip Culture, and sprouts rapidly.On the MS medium, the stem apex browning degree is serious, and poor growth has only the minority bud to launch, and the blade of expansion is also downright bad gradually.And stem apex inoculation has brown stain in the time of 2～3 days slightly on the WPM medium, generally change fresh culture after, can eliminate brown stain.After 1 week, stem apex is light green, expands, and bud is open in the time of 15 days, and Zhan Ye is branched out.When carrying out callus culture, the MS medium that macroelement reduces by half then more helps inducing, breed and breaking up of callus.
The WPM medium add variable concentrations BA (0.5～5.0mg/l) with NAA (0～0.2mg/l) can effectively induce stem apex and blade, petiole to produce indefinite bud.The optimum condition of stem apex adventitious bud inducing is NAA0.05mg/l, BA1.0mg/l, and on average each stem apex was cultivated through 1 month and can be produced 7.2 indefinite buds; The best hormone combinations of blade, petiole adventitious bud inducing is NAA 0.1mg/l, BA2.5mg/l, and wherein average each blade can be induced 6.0 indefinite buds, and on average each petiole can be induced 4.2 indefinite buds.Can induce two types callus from hypocotyl, at medium 1/2MS+2, the callus rate on 4-D 3.0mg/l+BA0.5mg/l+ sucrose 40 mg/l+ caseinhydrolysates (casein hydrolysace the is called for short CH) 1000mg/l is the highest.Inducing with sampling of male inflorescence axle, immature embryo callus is relevant period: the callus rate that obtains when wherein the male inflorescence axle is stretched in just by mixed bud is the highest, be 17.1%, the callus induction rate of immature embryo when developing into cotyledon shape embryo is the highest, is 12.9%.The callus in hypocotyl source differentiates indefinite bud on the differential medium that contains BA, ZT, NAA, the differentiation effect of ZT is good than BA.The paraffin wax section shows that indefinite bud originates from the meristematic tissue group that forms in interior of callus.
The indefinite bud that differentiates is on the WPM medium that the macroelement that adds 0.75～2.0mg/l IBA reduces by half, and about 2 weeks, rooting rate is more than 80%.
Shoot Tip Culture is a kind of valid approach for the fast numerous of select tree, and living seedling test was directly drawn materials from the select tree that grows up and carried out little numerous higher value that has on the basis in 1 year.In case after Shoot Tip Culture obtains aseptic seedling, utilize blade, the petiole evoking adventive bud of aseptic seedling to take place to increase reproduction coefficient again.And because without the callus stage, directly evoking adventive bud can reduce the generation of variant, for keeping the good characteristic of select tree, fine germplasm resources that guarantee is provided in the breeding fast.
The description of the drawings:
Accompanying drawing 1 is the influences of different hormone concentration combinations to Shoot Tip Culture bud quantity of differentiation;
Accompanying drawing 2 is different hormone concentration combinations influences to Shoot Tip Culture bud differentiation height;
Accompanying drawing 3 is different hormone concentration combinations influences to excised leaf, petiole differentiation;
Accompanying drawing 4 is the influences to the callus differentiation adventitious buds of three kinds of pretreatment modes.
Adopt the packaged technology of echinopodospora jamaicensis tissue-culturing quick-propagation provided by the present invention to carry out Chinese maple The exploitation of fragrant tissue cultivation and little propagating technology not only can be advanced on the basis that superior families is selected fast and efficiently The multiple target breeding research of row echinopodospora jamaicensis also can provide a kind of by the implant mass for sweetgum in production of forestry The method for culturing seedlings that cycle is short, breeding potential is high, with low cost.
Claims (7)
1. the method for an echinopodospora jamaicensis tissue-culturing quick-propagation is characterized in that, on the basis that select tree is selected, adopts 1 year living seedling stem apex of echinopodospora jamaicensis respectively, blade and petiole, and hypocotyl, male inflorescence axle and immature embryo carry out tissue culture.
2. the method for echinopodospora jamaicensis tissue-culturing quick-propagation as claimed in claim 1 is characterized in that, adopts 1 year living seedling stem apex of echinopodospora jamaicensis to carry out tissue culture.Stem apex growth and the used minimal medium of bud differentiation and proliferation are Woody P1ant Medium and Murashing and Skoogmedium, additional plant hormone is that concentration is the BA of 0～2.0mg/l, the NAA of 0～0.2mg/l removes the plant hormone combination that BA concentration is lower than NAA; The TDZ of the additional 0.01～0.1mg/l of WPM medium.
3. the method for echinopodospora jamaicensis tissue-culturing quick-propagation as claimed in claim 1 is characterized in that, the blade and the petiole that adopt 1 year living seedling stem-tip tissue of echinopodospora jamaicensis to turn out carry out tissue culture.It is Woody PlantMedium that excised leaf, petiole organ take place to cultivate used minimal medium, and additional plant hormone combination is the NAA of 0～0.1mg/l, the BA of 0.5～5.0mg/l; And the TDZ of additional 0.01mg/l.
4. the method for echinopodospora jamaicensis tissue-culturing quick-propagation as claimed in claim 1 is characterized in that, takes off plumular axis after employing echinopodospora jamaicensis seed is sprouted under aseptic condition, is seeded in the method for carrying out tissue culture on the MS/2 medium that adds hormone.
5. the method for echinopodospora jamaicensis tissue-culturing quick-propagation as claimed in claim 1 is characterized in that, adopts when mixed bud has just begun to expand outside male inflorescence stretches out mixed bud, gets the male inflorescence axle and is used for the method that callus induction carries out tissue culture; The used minimal medium of callus induction is MS/2.
6. the method for echinopodospora jamaicensis tissue-culturing quick-propagation as claimed in claim 1 is characterized in that, adopts at seed after 4～6 weeks of pollination, when the cotyledon type embryo occurring to seed, takes out rataria and is used for the method that callus induction carries out tissue culture; The used minimal medium of callus induction is MS/2.
7. as claim 4, the method for the echinopodospora jamaicensis tissue-culturing quick-propagation described in 5,6 is characterized in that, the hormone that callus induction is used is the 2.4-D of 0.5～3.0mg/l, the BA of 0～2.0mg/l.The hormone that adventitious bud inducing is used is the ZT of 0.5～1.0mg/l, the BA of 0.5～1.0mg/l and the NAA of 0.1mg/l.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01134054 CN1132512C (en) | 2001-10-18 | 2001-10-18 | Chinese sweetgum tissue culture and quick propagation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01134054 CN1132512C (en) | 2001-10-18 | 2001-10-18 | Chinese sweetgum tissue culture and quick propagation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1341351A true CN1341351A (en) | 2002-03-27 |
| CN1132512C CN1132512C (en) | 2003-12-31 |
Family
ID=4672190
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 01134054 Expired - Fee Related CN1132512C (en) | 2001-10-18 | 2001-10-18 | Chinese sweetgum tissue culture and quick propagation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1132512C (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823497A (en) * | 2012-09-19 | 2012-12-19 | 广东省林业科学研究院 | Clonal tissue culture breeding method of Liquidambar formosana Hance |
| CN103125388A (en) * | 2013-02-06 | 2013-06-05 | 中国林业科学研究院热带林业研究所 | Mytilaria laosensis tissue culturing method |
| CN103975856A (en) * | 2014-05-21 | 2014-08-13 | 中国长江三峡集团公司 | Tissue-culture rapid propagation method for inducing regeneration of liquidamba formosana hance embryoid |
| CN108719046A (en) * | 2017-04-13 | 2018-11-02 | 北京林业大学 | A kind of method of induction culturing hybrid sweetgum tetraploid |
| CN109673517A (en) * | 2019-02-20 | 2019-04-26 | 广西壮族自治区林业科学研究院 | A kind of method and culture medium improving sweetgum tissue culture shoot proliferation effect |
| CN110946073A (en) * | 2018-09-27 | 2020-04-03 | 东北林业大学 | Method for rapidly propagating fraxinus mandshurica embryonic cells |
| CN112616659A (en) * | 2020-12-14 | 2021-04-09 | 江苏省中国科学院植物研究所 | Method for somatic embryogenesis and plant regeneration of ilex davidii |
| CN113647318A (en) * | 2021-09-06 | 2021-11-16 | 北京农学院 | Method for promoting generation of new water-cultured Chinese chestnut roots |
| CN114365691A (en) * | 2022-02-21 | 2022-04-19 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of fortune purple maple |
| CN114467753A (en) * | 2022-02-21 | 2022-05-13 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of silvery deer maple |
-
2001
- 2001-10-18 CN CN 01134054 patent/CN1132512C/en not_active Expired - Fee Related
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823497B (en) * | 2012-09-19 | 2015-01-14 | 广东省林业科学研究院 | Clonal tissue culture breeding method of Liquidambar formosana hance |
| CN102823497A (en) * | 2012-09-19 | 2012-12-19 | 广东省林业科学研究院 | Clonal tissue culture breeding method of Liquidambar formosana Hance |
| CN103125388A (en) * | 2013-02-06 | 2013-06-05 | 中国林业科学研究院热带林业研究所 | Mytilaria laosensis tissue culturing method |
| CN103975856A (en) * | 2014-05-21 | 2014-08-13 | 中国长江三峡集团公司 | Tissue-culture rapid propagation method for inducing regeneration of liquidamba formosana hance embryoid |
| CN103975856B (en) * | 2014-05-21 | 2016-03-02 | 中国长江三峡集团公司 | A kind of tissue culture and rapid propagation method of inducing sweetgum somatic embryogenesis |
| CN108719046A (en) * | 2017-04-13 | 2018-11-02 | 北京林业大学 | A kind of method of induction culturing hybrid sweetgum tetraploid |
| CN108719046B (en) * | 2017-04-13 | 2021-12-24 | 北京林业大学 | Method for induced cultivation of hybrid liquidambar formosana tetraploid |
| CN110946073B (en) * | 2018-09-27 | 2021-04-16 | 东北林业大学 | Method for rapidly propagating fraxinus mandshurica embryonic cells |
| CN110946073A (en) * | 2018-09-27 | 2020-04-03 | 东北林业大学 | Method for rapidly propagating fraxinus mandshurica embryonic cells |
| CN109673517A (en) * | 2019-02-20 | 2019-04-26 | 广西壮族自治区林业科学研究院 | A kind of method and culture medium improving sweetgum tissue culture shoot proliferation effect |
| CN109673517B (en) * | 2019-02-20 | 2020-07-14 | 广西壮族自治区林业科学研究院 | Method and culture medium for improving tissue culture subculture proliferation effect of liquidambar formosana hance |
| CN112616659A (en) * | 2020-12-14 | 2021-04-09 | 江苏省中国科学院植物研究所 | Method for somatic embryogenesis and plant regeneration of ilex davidii |
| CN113647318A (en) * | 2021-09-06 | 2021-11-16 | 北京农学院 | Method for promoting generation of new water-cultured Chinese chestnut roots |
| CN114365691A (en) * | 2022-02-21 | 2022-04-19 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of fortune purple maple |
| CN114467753A (en) * | 2022-02-21 | 2022-05-13 | 德兴市荣兴苗木有限责任公司 | Tissue culture method of silvery deer maple |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1132512C (en) | 2003-12-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Etienne | Somatic embryogenesis protocol: coffee (Coffea arabica L. and C. canephora P.) | |
| CN1132512C (en) | Chinese sweetgum tissue culture and quick propagation method | |
| CN102907318A (en) | Rapid propagation of notoginseng regenerated plant by using bioreactor to cultivate notoginseng somatic embryos | |
| CN112931198A (en) | Preparation method of pineapple cold-resistant germplasm | |
| US7932086B2 (en) | Commercially viable process for in vitro mass culture of Jatropha curcas | |
| CN101548646B (en) | Method for rapidly propagating aralia elata through somatic embryo and secondary somatic embryogenesis | |
| CN1813525A (en) | Culture medium for anthurium andraeanum tissue culture and tissue culture seedling breeding method | |
| CN110741928B (en) | Walnut breeding method | |
| CN1140166C (en) | Somatic embryogenesis and plant regeneration technology for hybridized Chinese tuliptree | |
| KR101701494B1 (en) | Method of plant tissue culture for mass propagation of Corydalis turtschaninovii Besser. | |
| Agnihotri et al. | In vitro cloning of female and male Carica papaya through tips of shoots and inflorescences | |
| EP1817956A2 (en) | Commercially viable process for in-vitro mass culture of jatropha curcas | |
| CN102342246A (en) | Rhododendron decorum tissue-culture quick propagation method | |
| Mneney et al. | Clonal propagation of cashew (Anacardium occidentale L.) by tissue culture | |
| CN101785430A (en) | Tissue culture technology using Pinus massoniana isolated mature embryo as explant | |
| CN100355333C (en) | Bulbil konjak sprout breeding method | |
| Aguilar et al. | Production of cocoa plants (Theobroma cacao L.) via micrografting of somatic embryos | |
| CN1214708C (en) | Rapid breeding selection method of novel radish variety | |
| CN1180744A (en) | Method of tissue culture quick breeding for white cauliflower and green cauliflower cross breed new strain test-tube seeding | |
| CN1168376C (en) | Corn plant regenerating method and culture medium therefor | |
| Gomes et al. | Micropropagation of selected trees of Arbutus unedo L. through axillary shoot proliferation and somatic embryogenesis | |
| WO2000016610A1 (en) | A process for the induction of direct in vitro organogenesis in onion | |
| CN107873512A (en) | Anti- pine nematode black pine somatic embryo regeneration plant high efficiency breeding technology | |
| CN1572869A (en) | Media and methods for promoting maturation of conifer somatic embryos | |
| Tsegay et al. | Direct shoot organogenesis from juvenile cotyledon of Jatropha podagrica hook |
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 | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |