CN101911912A - Method for rapidly propagating jatrohpa curcas L. - Google Patents
Method for rapidly propagating jatrohpa curcas L. Download PDFInfo
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- CN101911912A CN101911912A CN2010102604953A CN201010260495A CN101911912A CN 101911912 A CN101911912 A CN 101911912A CN 2010102604953 A CN2010102604953 A CN 2010102604953A CN 201010260495 A CN201010260495 A CN 201010260495A CN 101911912 A CN101911912 A CN 101911912A
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Abstract
The invention relates to a method for rapidly propagating jatrohpa curcas L., which comprises the steps of: sterilizing a jatrohpa curcas L. seed, picking off the embryo of the sterilized jatrohpa curcas L. seed under the aseptic condition, inoculating to a culture medium, and culturing with light for 1 month; cutting a cotyledon or main leaf into small blocks, inoculating to an inducing culture medium prepared from 0.1-0.5mg/L of MS and TDZ, 0.01-0.1mg/L of BA, 30g/L of sucrose and 0.6-0.8% of agar, culturing for 2 weeks to generate callus, and continuing culturing for 2 weeks to differentiate an adventitious bud; transferring the adventitious bud into an enrichment culture medium prepared from 0.5-1.0mg/L of g1 and BA, 0.5-1.0mg/L of KT, 0.01-0.1mg/L of IBA, 1-5mg/L of AgNO3, 25-35g/L of sucrose and 0.6-0.8% of agar to enable tissue cultured crowd shoots to sprout from the adventitious bud; and cutting down a 2-3cm shoot, and inoculating into a rooting culture medium for rooting and culturing to obtain the tissue cultured rooted seedlings. The invention directly induces the cotyledon or tissue cultured seedling leaf to generate the callus and differentiate adventitious bud without replacing the culture medium, the differentiation frequency reaches 90 percent, and the induction process of the adventitious bud is greatly shortened.
Description
Technical field
The present invention relates to the little seeds of a tung oil tree method of breeding fast, especially a kind of blade that relates to tissue cultivating seedling is the raising technology of the plant nursery stock of induced material.
Background technology
The little seeds of a tung oil tree (
Jatropha curcasL.) crying Jatropha curcas again, is Euphorbiaceae Jatropha curcas platymiscium.Perennial, dungarunga or shrub.Mainly be distributed in the torrid zone and subtropical zone, China in Guangdong, Guangxi, Yunnan, Guizhou, Fujian, Hainan etc. economize widely plantation, or wildly be in groups.Its seed oil content height can reach 40 ~ 60%, is the universally acknowledged promising bioenergy plant that is.Its grease can become diesel oil after extraction and sex change.This biodiesel is biodegradable, and environmental sound is more clean and efficient than mineral diesel.Under the situation that the energy is in short supply day by day at present, Jatropha curcas is as a kind of reproducible bioenergy, extremely people's attention.Many countries have carried out the research of this respect, the great scientific research and development project that development utilization and the research of Jatropha curcas are listed more for a long time by the Chinese government in country.In addition, Jatropha curcas still is a kind of medicinal plant, and multiple medicinal ingredient is arranged, and also is the pesticide material that good insecticidal effect is arranged.Drought-resistant, the soil depletion of Jatropha curcas, breeding is convenient, but seed sowing, but cottage propagation is planted easyly, can plant in a large number at the uncultivated area band, is to improve the ecological environment, develop the desert, the fabulous seeds of increasing peasant income.
At present, utilize tissue culture technique to carry out report more (Qin Hong, Song Songquan, the Long Chunlin etc. of the quick proliferation of the little seeds of a tung oil tree.The tissue culture of the little seeds of a tung oil tree and plant regeneration [J]. Yunnan plant research, 2006,28(6): 649-652.; Jatropha curcas stem section cultured in vitro and breeding research [J] fast. the Guangxi agricultural science, 2006,37(3): 221-223.; Lu Weida, Wei Qin, Tang Lin etc.Inducing and breeding [J] fast of Jatropha curcas callus. use and the environmental organism journal 2003,9 (2): 127-130.; Lin Juan, Tang Lin displays.Tissue culture of Jatropha curcas and plant regeneration [J]. Plant Physiology Communications, 2002,38(3): 252. etc.), but there are the following problems:
1, all adopts basic element of cell division 6-benzyladenine (6-BA) and growth hormone indolebutyric acid (IBA) or the combination callus induction of methyl (NAA) variable concentrations and the differentiation of indefinite bud.Formation required time from the callus induction to the indefinite bud is long, about 6 ~ 12 weeks;
2, in the above report, all adopt different explant (leaflet tablet of cotyledon, hypocotyl, plumule, earsh and petiole, stem section etc.), all can evoked callus, but that callus is divided into the frequency difference of indefinite bud is very big, is 0 ~ 80%;
3, the bud growth rate is slow, can not satisfy the demand that batch production is produced.
Adopt above method, carry out the callus induction seedling differentiation, individual reproduction speed is not ideal enough, for addressing this problem, has the researcher to carry out promoting the way of Jatropha curcas axillalry bud branch to breed achieving success (Leeization, Ceng Ni, Jia Yongjiong etc. fast.The fast numerous and root induction [J] of the short axillalry bud branch of Jatropha curcas. Sichuan University's journal, 2006,43(5): 1116-1120).This approach, though making the speed of bud propagation accelerates greatly, but still can't avoid making the accumulation of plant corpus inner cell mitogen because of subculture repeatedly, the situation that causes tissue cultivating seedling growth potential to die down takes place, also can make the tissue cultivating seedling of acquisition have the difficult problem of taking root, the bad tissue cultivating seedling of unrooted or root system development is difficult to transplant survival.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of little seeds of a tung oil tree method of breeding fast, has desirable high differentiation frequency.
The present invention solves the problems of the technologies described above the technical scheme of being taked: a kind of little seeds of a tung oil tree are the method for breeding fast, carries out tissue culture by following step in regular turn:
The first step: after the seed disinfection with the little seeds of a tung oil tree, strip embryo under aseptic condition, be inoculated in the medium of 1/2MS+ sucrose 25 ~ 35g/L+0.6 ~ 0.8% agar, illumination cultivation was carried out 28 ~ 35 days in pH5.8 ~ 6.2, obtained aseptic seedling;
Second step: the cotyledon or the true leaf of aseptic seedling are cut into 0.4 ~ 0.6cm
2Fritter, be inoculated in the inducing culture, described inducing culture is MS+TDZ0.1 ~ 0.5mg/L+IBA0.01 ~ 0.1mg/L+ sucrose 25 ~ 35g/L+0.6 ~ 0.8% agar, pH5.8 ~ 6.2, cultivate and produced callus in 12 ~ 16 days, continue cultivation and differentiated indefinite bud in 12 ~ 16 days;
The 3rd step: change indefinite bud over to proliferated culture medium and carry out the shoot proliferation cultivation, described proliferated culture medium is gl+BA0.5 ~ 1.0mg/L+KT0.5 ~ 1.0mg/L+IBA0.01 ~ 0.1mg/L+AgNO
31 ~ 5mg/L+ sucrose, 25 ~ 35g/L+0.6 ~ 0.8% agar, pH5.8 ~ 6.2, generation group training clump bud;
The 4th step: the bud that will organize 2 ~ 3cm in the training clump bud downcuts, insert the root of hair medium and carry out culture of rootage, the root of hair medium is MS+NAA0.5 ~ 1.5 mg/L+ sucrose, 15 ~ 25g/L+0.6 ~ 0.8% agar, pH5.8 ~ 6.2, change over to after 3 ~ 7 days in the medium of MS+ sucrose 15 ~ 25g/L+0.6 ~ 0.8% agar, took root through 10 ~ 20 days, cultivate 28 ~ 35 days acquisition groups and train the seedling of taking root, transplant.
On the basis of such scheme, the group training of the getting robust growth blade of seedling of taking root, repeating step two be to step 4, to substitute the group training clump bud that dies down through the growth impetus behind the subculture repeatedly.
On the basis of such scheme, described seed disinfection method is: soaked in 70 ~ 80% alcohol 2 ~ 3 minutes, under aseptic condition, peel off and used 70 ~ 80% alcohol disinfecting behind kind of the skin again 2 ~ 3 minutes, then with 0.05% mercuric chloride sterilization 15 ~ 25 minutes, finish the back with aseptic water washing at least 4 times, blot redundant moisture with sterile gauze at last.
On the basis of such scheme, described seed disinfection method is: the condition of described illumination cultivation is: 25 ± 2 ℃ of culturing room's temperature, intensity of illumination 1500 ~ 2000LX, 10 ~ 14 hours every days of light application time.
The invention has the beneficial effects as follows:
1, with basic element of cell division Thidiazuron (TDZ) 0.1 ~ 0.5mg/L and growth hormone indolebutyric acid (IBA) 0.01 ~ 0.1mg/L combination, directly induce cotyledon or tissue cultivating seedling blade to produce callus and differentiate indefinite bud, the process of callus differentiation indefinite bud does not need to change culture medium prescription.The method can differentiate indefinite bud in 1 month, and differentiation frequency reaches 90%, had shortened the process of inducing of indefinite bud greatly;
2, adopt above same recipe, also can make the take root blade of seedling of group training differentiate indefinite bud, behind enrichment culture, the alternative group training clump bud that dies down through growth potential behind the subculture repeatedly, so form production cycle, produce high-quality group endlessly and cultivate seedling.
Embodiment
A kind of little seeds of a tung oil tree are the method for breeding fast, carries out tissue culture by following step in regular turn:
The first step: small idesia was soaked 2 minutes in 75% alcohol, under aseptic condition, peel off and used 75% alcohol disinfecting behind kind of the skin again 2 minutes, with 0.05% mercuric chloride sterilization 20 minutes, back aseptic water washing 6 times that finish were blotted redundant moisture with sterile gauze at last then; Under aseptic condition, strip embryo, be inoculated in the medium of 1/2MS+ sucrose 30g/L+0.6-0.8% agar pH5.8 ~ 6.2 then, carry out illumination cultivation, 25 ± 2 ℃ of culturing room's temperature, intensity of illumination 1500 ~ 2000LX, 10 ~ 14 hours every days of light application time, cultivated for 2 weeks, obtain aseptic seedling;
Second step: the cotyledon or the true leaf of aseptic seedling are cut into 0.5cm
2Fritter, be inoculated in the inducing culture, described inducing culture is MS+TDZ0.1 ~ 0.5mg/L+IBA0.01 ~ 0.1mg/L+ sucrose 30g/L+0.6 ~ 0.8% agar, pH5.8 ~ 6.2 are cultivated and 2 weeks were produced callus, healing rate 100%, cultivated for 2 weeks again, differentiate indefinite bud, differentiation frequency 90%, average every explant can produce 6.3 indefinite buds;
The 3rd step: change indefinite bud over to proliferated culture medium and carry out the shoot proliferation cultivation, described proliferated culture medium is gl+BA0.5 ~ 1.0mg/L+KT0.5 ~ 1.0mg/L+IBA0.01 ~ 0.1mg/L+AgNO
31 ~ 5mg/L+ sucrose 30g/L+0.6 ~ 0.8% agar, pH5.8 ~ 6.2, generation group training clump bud, subculture 1 time, the propagation ratio is 1:2;
The 4th step: the bud of 2 ~ 3cm in the group training clump bud of shoot proliferation incubation acquisition is downcut, insert the root of hair medium, described root of hair medium is MS+NAA0.5 ~ 1.5 mg/L+ sucrose 20g/L+0.6 ~ 0.8% agar, pH5.8 ~ 6.2, change over to after 3 ~ 7 days in the medium of MS+ sucrose 20g/L+0.6 ~ 0.8% agar, took root through 10 ~ 20 days, rooting rate can be transplanted in 90%, 1 month.
The group training of the getting robust growth blade of seedling of taking root repeated for second step to the 4th step, to substitute the group training clump bud that dies down through the growth impetus behind the subculture repeatedly.
Claims (4)
1. the little seeds of a tung oil tree method of breeding fast, carry out tissue culture by following step in regular turn:
The first step: after the seed disinfection with the little seeds of a tung oil tree, strip embryo under aseptic condition, be inoculated in the medium of 1/2MS+ sucrose 25～35g/L+0.6～0.8% agar, illumination cultivation was carried out 28～35 days in pH5.8～6.2, obtained aseptic seedling;
Second step: the cotyledon or the true leaf of aseptic seedling are cut into 0.4～0.6cm
2Fritter, be inoculated in the inducing culture, described inducing culture is MS+TDZ0.1～0.5mg/L+IBA0.01～0.1mg/L+ sucrose 25～35g/L+0.6～0.8% agar, pH5.8～6.2, cultivate and produced callus in 12～16 days, continue cultivation and differentiated indefinite bud in 12～16 days;
The 3rd step: change indefinite bud over to proliferated culture medium and carry out the shoot proliferation cultivation, described proliferated culture medium is gl+BA0.5～1.0mg/L+KT0.5～1.0mg/L+IBA0.01～0.1mg/L+AgNO
31～5mg/L+ sucrose, 25～35g/L+0.6～0.8% agar, pH5.8～6.2, generation group training clump bud;
The 4th step: the bud that will organize 2～3cm in the training clump bud downcuts, insert the root of hair medium and carry out culture of rootage, the root of hair medium is MS+NAA0.5～1.5 mg/L+ sucrose, 15～25g/L+0.6～0.8% agar, pH5.8～6.2, change over to after 3～7 days in the medium of MS+ sucrose 15～25g/L+0.6～0.8% agar, took root through 10～20 days, cultivate 28～35 days acquisition groups and train the seedling of taking root, transplant.
2. the little seeds of a tung oil tree according to claim 1 are the method for breeding fast, it is characterized in that: the group training of the getting robust growth blade of seedling of taking root, repeated for second step to the 4th step, to substitute the group training clump bud that dies down through the growth impetus behind the subculture repeatedly.
3. the little seeds of a tung oil tree according to claim 1 are the method for breeding fast, it is characterized in that: described seed disinfection method is: soaked in 70～80% alcohol 2～3 minutes, under aseptic condition, peel off and used 70～80% alcohol disinfecting behind kind of the skin again 2～3 minutes, then with 0.05% mercuric chloride sterilization 15～25 minutes, finish the back with aseptic water washing at least 4 times, blot redundant moisture with sterile gauze at last.
4. the little seeds of a tung oil tree according to claim 1 are the method for breeding fast, it is characterized in that: described seed disinfection method is: the condition of described illumination cultivation is: 25 ± 2 ℃ of culturing room's temperature, intensity of illumination 1500～2000LX, 10～14 hours every days of light application time.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102524064A (en) * | 2011-12-19 | 2012-07-04 | 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 |
| CN105165615A (en) * | 2015-09-26 | 2015-12-23 | 中南民族大学 | Culturing method for sunflower callus tissue |
| CN106212288A (en) * | 2016-08-24 | 2016-12-14 | 华南农业大学 | A kind of tissue culture propagation method of Machilus pauhoi |
| CN106577277A (en) * | 2016-11-23 | 2017-04-26 | 云南云投生态环境科技股份有限公司 | Callus induction, differentiation and plant regeneration method of plukenetia volubilis linneo |
| CN106857247A (en) * | 2017-01-16 | 2017-06-20 | 华南农业大学 | A kind of method that Jatropha curcas aseptic seedling respectively organizes callus induction |
| CN107771673A (en) * | 2017-11-14 | 2018-03-09 | 广西壮族自治区药用植物园 | The rapid propagation method of Chinese tallow tree callus seedling |
| CN113892432A (en) * | 2021-11-22 | 2022-01-07 | 合肥师范学院 | Method for directly regenerating barbadosnut leaf |
-
2010
- 2010-08-24 CN CN2010102604953A patent/CN101911912A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102524064A (en) * | 2011-12-19 | 2012-07-04 | 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 |
| CN105165615A (en) * | 2015-09-26 | 2015-12-23 | 中南民族大学 | Culturing method for sunflower callus tissue |
| CN106212288A (en) * | 2016-08-24 | 2016-12-14 | 华南农业大学 | A kind of tissue culture propagation method of Machilus pauhoi |
| CN106212288B (en) * | 2016-08-24 | 2018-07-31 | 华南农业大学 | A kind of tissue culture propagation method of Machilus pauhoi |
| CN106577277A (en) * | 2016-11-23 | 2017-04-26 | 云南云投生态环境科技股份有限公司 | Callus induction, differentiation and plant regeneration method of plukenetia volubilis linneo |
| CN106857247A (en) * | 2017-01-16 | 2017-06-20 | 华南农业大学 | A kind of method that Jatropha curcas aseptic seedling respectively organizes callus induction |
| CN106857247B (en) * | 2017-01-16 | 2019-11-08 | 华南农业大学 | A kind of method that Jatropha curcas aseptic seedling respectively organizes callus induction |
| CN107771673A (en) * | 2017-11-14 | 2018-03-09 | 广西壮族自治区药用植物园 | The rapid propagation method of Chinese tallow tree callus seedling |
| CN113892432A (en) * | 2021-11-22 | 2022-01-07 | 合肥师范学院 | Method for directly regenerating barbadosnut leaf |
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Application publication date: 20101215 |