CN100407903C - Fast teething and reproduction for Mafeng tree - Google Patents
Fast teething and reproduction for Mafeng tree Download PDFInfo
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- CN100407903C CN100407903C CNB200610020449XA CN200610020449A CN100407903C CN 100407903 C CN100407903 C CN 100407903C CN B200610020449X A CNB200610020449X A CN B200610020449XA CN 200610020449 A CN200610020449 A CN 200610020449A CN 100407903 C CN100407903 C CN 100407903C
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Abstract
The present invention provides a method for the fast propagation of Jatropha curcas outgrowths. In the method, Jatropha curcas sprouts sterilized are put in a fast propagation culture medium with the pH value of 4.8 to 6.6, and the culture is carried out for 25 to 35 days under the conditions of temperature of 25 to 35 DEG C, the illumination intensity of 1500 to 3000 lx and the illumination time of 8 to 15 h/d to induce the propagation of outgrowths. Meanwhile, the present invention provides two methods for the fast propagation of Jatropha curcas rooting seedlings. In the methods, firstly, sterile outgrowth incisions are moved into an MS basic culture medium or a 1/2MS basic culture medium after being soaked in a sterile growth hormone, or outgrowths are directly put in an MS basic culture medium or a 1/2MS basic culture medium with active carbon to achieve rooting culture. The present invention has the advantages of high speed for outgrowth propagation, high average budding number and high rooting rate for the propagation of rooting seedlings, and the highest rooting rate can reach 100%. The present invention can provide good rooting seedlings for undeveloped-mountain afforestation and the wide planting of Jatropha curcas.
Description
One, technical field
The invention belongs to the micropropagation of plants technical field, be specifically related to the method for a kind of quick breeding bud of Cortex jatrophae and the seedling of taking root.
Two, background technology
Jatropha curcas (Jatropha curcas L.) belongs to Euphorbiaceae (Euphorbiaceae) leprosy Pterostyrax (Jatropha) machaka or dungarunga, originates in tropical America, is distributed in the torrid zone, the world and subtropical zone.China cultivation or semi-wild in Taiwan, ground such as Guangdong, Guangxi, Sichuan, Guizhou, Hainan and Yunnan.The comprehensive utilization value height of Jatropha curcas, its leaf can be treated rheumatism, venereal disease, angina pectoris; Branch milk then has better curative effect to wart, skin trauma etc.; The branch extract has the effect of anti HIV-1 virus; Root can be used for loosing becoming silted up and subsides a swelling, stops blooding; Seed can be used as cathartic; Toxalbumin from jatropha curcas seed extracts has stronger inhibitory action to tumour cell; Jatropha curcas seed also can be used for the extermination of disease and insect pest of plant, also is a kind of oil crop with important economic worth simultaneously, and its kind benevolence oil-containing is about 50%, and seed oil is processed the substitute that just can be used as diesel oil slightly.Even have and report and need not to improve under the diesel engine or the situation of pre-heating fuel, can be directly with the Jatropha curcas oil and diesel oil mixed combustion of 40-50%.And Jatropha curcas is barren anti-dried morning, is the good seeds of dry-hot valley area afforestation, therefore has exploitation prospect widely.
At present, the breeding and culturing means of Jatropha curcas mainly adopt traditional planting seed kind method for planting and cuttage planting method (Lin Juan, weekly selection is enclosed, Tang Kexuan etc. Jatropha curcas plant resources research overview [J]. the tropical and subtropical zone Botany Gazette, 2004,12 (3): 285-290.; Shi Zongming, Li Yun, Luo Yuxing etc. the development and use of the little seeds of a tung oil tree of energy-source plant and cultivation research. Yunnan Normal University's journal, 1992,12 (2): 31-38.).There are the following problems for these two kinds of methods: (1) realizes that by self-sow the speed of Jatropha curcas individual reproduction is slow.Generally breeding from planting seed to ripe result needed for 3 years again, also needed year at least even cuttage cultivation is grown in control environment.(2) realize that by self-sow the Jatropha curcas individual reproduction is limited by season and region easily.As plant Jatropha curcas need be before rainy season, because of moisture to the survival influence of Jatropha curcas greatly.(3) natural propagation is unfavorable for the maintenance of merit.
In order to solve the problem that traditional breeding and culturing means exist, there is the researcher to carry out the research of relevant Jatropha curcas callus of induce seedling differentiation, and (the Lin Juan that succeeds, Tang Lin, display. tissue culture of Jatropha curcas and plant regeneration. Plant Physiology Communications, 2005,38 (3): 252.; Lu Weida, Wei Qin, Tang Lin etc. inducing and breeding fast of Jatropha curcas callus. use and the environmental organism journal 2003,9 (2): 127-130.).Though the mode of callus of induce seedling differentiation is than traditional breeding and culturing mode, improved individual reproduction speed, avoided individual reproduction to be subjected to season and region limitations affect easily, but the individual reproduction speed that improves is desirable not enough, and this class differentiation seedling aberration rate height may be lost the part merit in atomization.In addition, because the process of callus of induce seedling differentiation has used the basic element of cell division of high concentration to induce, so make the tissue culturing seedling of its acquisition have the problem of the difficulty of taking root, and the bad tissue cultivating seedling of root system development is in case transplanting will soon be dead in soil, cause it to be grown on the medium, thereby can not provide the good seedling of taking root for Jatropha curcas carries out afforestation.
Three, summary of the invention
At the problem that prior art exists, first purpose of the present invention provides a kind of method of quick breeding bud of Cortex jatrophae.
Second purpose of the present invention provides the take root method of seedling of a kind of quick breeding Jatropha curcas.
The 3rd purpose of the present invention provides the take root method of seedling of the another kind of Jatropha curcas of breeding fast.
The method of quick breeding bud of Cortex jatrophae provided by the invention, it is characterized in that this method is that the Jatropha curcas bud after the sterilization is placed the pH value is on 4.8～6.6 the fast propagating culture medium, and at cultivation temperature 25-35 ℃, intensity of illumination 1500-3000lx, light application time 8-15h/d cultivate down can induce in 25-35 days and breed young shoot.
The young shoot that cutting-out breeds can place to cultivate on this medium to produce more young shoot repeatedly.
Used fast propagating culture medium is in the MS medium in the said method, adds and counts with the volume of MS medium: 0.5～2.5mg/L basic element of cell division and 0.01～0.1mg/L growth hormone are formed.The wherein used basic element of cell division is any in 6-benzyladenine, the zeatin; Used growth hormone is any in indolebutyric acid, naa, the heteroauxin.
The pH value is regulated the available hydrogen sodium oxide molybdena or hydrochloric acid carries out.
In order to promote young shoot better to grow, in fast propagating culture medium, also can add the silver nitrate of counting 1～10mg/L with the volume of MS medium.
First kind of quick take root method of seedling of breeding Jatropha curcas provided by the invention, it is characterized in that this method is is to soak 0.5～3h in 100～500mg/L growth hormone with aseptic young shoot otch in aseptic concentration earlier, move to then in MS or the 1/2MS minimal medium, at cultivation temperature 25-35 ℃, intensity of illumination 1500-3000lx, light application time 8-15h/d cultivate down can become after 30-40 days to reach and transplant the seedling of taking root that requires.
Fast the breeding Jatropha curcas take root in the seedling-growing method used growth hormone be indolebutyric acid, naa, 2,4 dichlorophenoxyacetic acid (2,4-D) in any.
Another kind provided by the invention is bred the take root method of seedling of Jatropha curcas fast, it is characterized in that this method is young shoot directly to be placed added MS or the 1/2MS minimal medium of counting 0.1～3.0mg/L active carbon with the volume of medium, at cultivation temperature 25-35 ℃, intensity of illumination 1500-30001x, light application time 8-15h/d cultivate down can become after 30-40 days to reach and transplant the seedling of taking root that requires.
The present invention has following advantage:
1, owing to used fast propagating culture medium in the method for quick breeding bud of Cortex jatrophae provided by the invention is in the MS medium, the basic element of cell division and growth hormone have been added, thereby just can on explant, cultivate young shoot about one month, not only fast than the speed of conventional method individual reproduction, and also faster than callus of induce seedling differentiation method reproduction speed (because of there not being the cultivating process of callus).
2, owing to be in the MS medium in the used fast propagating culture medium in the method for quick breeding bud of Cortex jatrophae provided by the invention, the basic element of cell division and growth hormone have been added, thereby the number that on average sprouts is high, the highest number that on average sprouts can be up to 6.15, and only need few material just can turn out many strain young seedling of Jatropha in the short time.
3, because the method for quick breeding bud of Cortex jatrophae provided by the invention is to carry out, belong to in-house operation on medium, thereby be not subjected to the influence of season and region restriction, can breed whenever and wherever possible.
4, because the method for quick breeding bud of Cortex jatrophae provided by the invention belongs to the enhanced axillary branching and rooting technique category, thereby the branch that is produced by its young shoot propagation is to be grown by the nutrition meristematic tissue that exists originally, keep former clonal genotype and phenotype, avoid natural propagation to be unfavorable for the maintenance of merit and callus of induce seedling differentiation breeding aberration rate height, in atomization, may lose the defective of part merit.
5, adopted elder generation that aseptic young shoot otch has been carried out immersion treatment in the asepsis growth element owing to the invention provides the take root method of seedling of quick breeding Jatropha curcas, the technical measures of active carbon perhaps in minimal medium, have been added, thereby not only solve the difficult problem of taking root that prior art tissue culturing seedling exists, improve the rooting rate of inducing the Jatropha curcas tissue cultivating seedling of generation by high concentration of cytokinin significantly, reach as high as 100%, and ratio is also high without the rooting rate of the bud of Cortex jatrophae of the self-sow of crossing basic element of cell division influence, thereby can carry out afforestation for Jatropha curcas, extensively plantation provides the good seedling of taking root.
Four, embodiment
Below by embodiment the present invention is specifically described; be necessary to be pointed out that at this following examples only are used for that the invention will be further described; can not be interpreted as limiting the scope of the invention; the person skilled in the art in this field makes some nonessential improvement and adjustment according to the content of the invention described above to the present invention, still belongs to protection scope of the present invention.
Embodiment 1～6, comparative example 1
This group embodiment and comparative example all adopt the Jatropha curcas stem apex of self-sow to breed fast.
Sterilization treatment intercepts the Jatropha curcas stem apex 1～2cm of self-sow respectively by each embodiment and comparative example inoculation quantity, puts into 70% alcohol and shakes 1min, discards alcohol, uses 0.1%HgCl again
2Solution disinfection 7min discards HgCl
2, use aseptic water washing then 5 times, with the remaining HgCl of thorough flush away
2
The preparation fast propagating culture medium is in the MS medium of (1)～(9) numbering, press basic element of cell division 6-benzyladenine (BA), growth hormone indolebutyric acid (IBA) and the silver nitrate of table 1 interpolation respectively, and its pH value is adjusted to 5.8 with the volume metering of MS medium.
The moisture on stem apex surface after sterilization treatment is blotted in breeding fast, cut unnecessary part, only keep the tender part of most advanced and sophisticated children, direction according to self-sow moves into respectively in the fast propagating culture medium of (1)～(9) numbering then, in cultivation temperature is 25 ℃, and illuminance 1500lx cultivated 30 days under the light application time 12h/d condition, measure the young shoot amount of inducing, the results are shown in Table 1.
Table 1
The result shows: the collocation of BA and IBA helps the numerous soon of Jatropha curcas stem apex, adds an amount of AgNO in the medium
3Can prevent brownization, reduce lopsided seedling, promote the differentiation of bud.
Embodiment 9～17
This group embodiment all adopts the aseptic seedling of being cultivated by hormonal stimulation to breed fast.
Sterilization treatment is by the quantity of each embodiment inoculation, and the kind shell of strip off jatropha curcas seed takes out embryo wherein, puts into 70% alcohol and shakes 1min, discards alcohol, uses 0.1%HgCl again
2Solution disinfection 7min discards HgCl
2, use aseptic water washing then 3 times, with the remaining HgCl of thorough flush away
2, after adding sterile water again and soaking 48h, outwell sterile water.Operate once again by above-mentioned sterilization process.
Cultivate aseptic seedling and blot the moisture of after sterilization treatment, planting the benevolence surface, cut kind of a benevolence (endosperm) with blade along the longitudinal axis, strip out two white cotyledons (two slice, thin piece leaf bases are entrained with embryo), cotyledon is connected embryo, and to be seeded in the pH value be 5.8, and added with culture volume and counted in the MS medium of 0.1mg/L 6-benzyladenine (BA), in cultivation temperature is 28 ± 1 ℃, and illuminance 2000lx cultivates under the condition of light application time 12h/d and promptly obtained aseptic seedling in 20-30 days.
The preparation fast propagating culture medium is in the MS medium of (10)～(17) numbering, press basic element of cell division 6-benzyladenine (BA), growth hormone naa (NAA) and the silver nitrate of table 2 interpolation respectively, and its pH value is adjusted to 5.8 with the volume metering of MS medium.
Fast the aseptic seedling of sprouting is got in breeding, and plumule is downcut, be seeded in respectively (10)～fast propagating culture medium of (17) numbering in, in cultivation temperature is 28 ± 1 ℃, and illuminance 2000lx cultivated 30 days under the light application time 12h/d condition, measure the young shoot amount of inducing, the results are shown in Table 2.
Table 2
The result shows: the collocation of BA and NAA helps the numerous soon of Jatropha curcas stem apex, adds an amount of AgNO in the medium
3Can prevent brownization, reduce lopsided seedling, promote the differentiation of bud.
Embodiment 18
Will by embodiment 9～17 describe through sterilization treatment, cultivate the plumule that the aseptic seedling operation cultivates the aseptic seedling of sprouting and downcut, inoculate 20 by MS+2.0mg/L zeatin+0.05mg/L heteroauxin (IAA)+5mg/L AgNO
3In the fast propagating culture medium of forming, the medium pH value is 4.8, and is 30 ℃ in cultivation temperature, and illuminance 2000lx cultivates after 25 days under the condition of light application time 8h/d, and it is 2.70 that axillalry bud is on average induced number.
Embodiment 19
Will by embodiment 9～17 describe through sterilization treatment, cultivate the plumule that the aseptic seedling operation cultivates the aseptic seedling of sprouting and downcut, inoculate 20 by MS+2.0mg/L zeatin+0.05mg/L heteroauxin (IAA)+5mg/L AgNO
3In the fast propagating culture medium of forming, the medium pH value is 6.6, and is 35 ℃ in cultivation temperature, and illuminance 3000lx cultivates after 35 days under the condition of light application time 15h/d, and it is 3.78 that axillalry bud is on average induced number.
Embodiment 20～25, comparative example 2,3
This group embodiment and comparative example are for cultivating the take root example of seedling of Jatropha curcas.
During the young shoot length to 1 that stand-by the inventive method is induced～2cm, can downcut young shoot and directly move to by table 3 and added in the root media of active carbon, and press the given breeding condition root induction of table 3, the results are shown in Table 3.
The otch of the young shoot that also another part is downcut soaks a period of time earlier, and then moves in MS or the 1/2MS medium in the aseptic high concentration growth element by table 3 configuration simultaneously, presses the given breeding condition root induction of table 3, the results are shown in Table 3.
The pH value of medium all is 5.8.
Comparative example 2 from table 3 as can be seen, though can spontaneously take root without the bud of Cortex jatrophae of crossing the basic element of cell division and influence self-sow, rooting rate is not high comparatively speaking, only is 65%.And from comparative example 3 as can be seen, induce the aseptic young shoot of generation by MS+2.0mg/L BA+0.05mg/L IBA medium, and under situation, lost spontaneous ability of taking root without any processing, can not spontaneously take root at all.Yet according to the seedling of taking root that method provided by the invention is cultivated, rooting rate can be greatly improved.
Table 3
Claims (4)
1. quick method of breeding Jatropha curcas, it is characterized in that this method is the Jatropha curcas stem apex with the self-sow after the sterilization, or the aseptic seedling of cultivating by hormonal stimulation, placing the pH value is on 4.8～6.6 the fast propagating culture medium, and at cultivation temperature 25-35 ℃, intensity of illumination 1500-3000lx, light application time 8-15h/d cultivates down promptly to induce in 25-35 days and breeds young shoot, used fast propagating culture medium is in the MS medium, adds and counts with the volume of MS medium: the basic element of cell division of 0.5～2.5mg/L, 0.01 the growth hormone of～0.1mg/L and the silver nitrate of 1～10mg/L;
Then the young shoot that induces is directly placed MS or 1/2MS medium, in medium, be added with the active carbon of counting 0.1～3.0mg/L with the volume of medium, at cultivation temperature 25-35 ℃, intensity of illumination 1500-3000lx, the light application time 8-15h/d degree of taking root after 30-40 days of cultivating down reaches the requirement of transplanting; Perhaps, the young shoot that induces is downcut, is to soak 0.5～3h in the growth hormone of 100～500mg/L with otch in the concentration of sterilization, move to then in MS or the 1/2MS medium, at cultivation temperature 25-35 ℃, intensity of illumination 1500-3000lx, the light application time 8-15h/d degree of taking root after 30-40 days of cultivating down reaches the requirement of transplanting.
2. the method for quick breeding Jatropha curcas according to claim 1 is characterized in that the basic element of cell division used in the fast propagating culture medium is any in 6-benzyladenine, the zeatin.
3. the method for quick breeding Jatropha curcas according to claim 1 and 2 is characterized in that growth hormone used in the fast propagating culture medium is any in indolebutyric acid, naa, the heteroauxin.
4. the method for quick breeding Jatropha curcas according to claim 1, the growth hormone that it is characterized in that soaking young shoot are any in indolebutyric acid, naa, the 2,4 dichlorophenoxyacetic acid.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101574058B (en) * | 2009-06-16 | 2011-11-16 | 四川大学 | Barbadosnut plantlet tissue culture rapid propagation and rooting method |
| CN103283592A (en) * | 2013-04-17 | 2013-09-11 | 华南农业大学 | Method for direct regeneration of adventitious bud from Jatropha curcas petiole explant |
| CN103430841A (en) * | 2013-08-02 | 2013-12-11 | 华南农业大学 | Method for promoting jatropha curcas explant to regenerate adventitious buds |
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| CN101057557B (en) * | 2007-06-01 | 2010-05-19 | 中山大学 | Non-tube quick reproduction method for seedings of jatropha curcas L. |
| CN101103692B (en) * | 2007-08-03 | 2010-05-19 | 广东金浩能源有限公司 | Introduction and cultivation method of high oil content jatropha curcas |
| CN101361457B (en) * | 2008-09-27 | 2010-11-17 | 西南林学院 | Jatropha regeneration method using excised leaf |
| CN102405832B (en) * | 2011-08-18 | 2014-03-05 | 嘉汉林业(广州)有限公司 | Method for rapid propagation of Jatropha curcas |
| CN102524064B (en) * | 2011-12-19 | 2014-04-16 | 普罗米绿色能源(深圳)有限公司 | Rejuvenation and rooting culturing method for genetic transformation seedlings of Jatropha curcas |
| CN103416307B (en) * | 2013-08-05 | 2015-05-20 | 四川大学 | Rapid propagation method for directly inducing buds by using jatropha curcas L. seed leaf regenerated roots |
| CN103749295B (en) * | 2013-12-31 | 2015-12-09 | 四川大学 | A kind of method of Jatropha curcas regrowth extraneous root |
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2006
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101574058B (en) * | 2009-06-16 | 2011-11-16 | 四川大学 | Barbadosnut plantlet tissue culture rapid propagation and rooting method |
| CN103283592A (en) * | 2013-04-17 | 2013-09-11 | 华南农业大学 | Method for direct regeneration of adventitious bud from Jatropha curcas petiole explant |
| CN103283592B (en) * | 2013-04-17 | 2014-11-12 | 华南农业大学 | Method for direct regeneration of adventitious bud from Jatropha curcas petiole explant |
| CN103430841A (en) * | 2013-08-02 | 2013-12-11 | 华南农业大学 | Method for promoting jatropha curcas explant to regenerate adventitious buds |
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