CN102893864B - Rhus chinensis adventitious bud high frequency plant regeneration method - Google Patents
Rhus chinensis adventitious bud high frequency plant regeneration method Download PDFInfo
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Abstract
The invention discloses a Rhus chinensis adventitious budr high frequency plant regeneration method. The method has high regeneration efficiency, fast propagation speed and high average germination rate up to 95%; and the obtained sprout has good rooting rate, and each plant produces about 4-8 robust root systems. The present invention has proved that rapid propagation of Rhus chinensis seedling can be realized without through a seed seedling method or a root pressure method, so as to save time, release time limits, greatly improve reproductive efficiency, and rapidly provide sufficient seedlings for construction of Rhus chinensis economic forest; at the same time, the high efficiency regeneration method lays good foundation for molecular improvement especially transgenic technology of Rhus chinensis.
Description
Technical field
The invention belongs to plant biotechnology field, be specifically related to a kind of method of Chinese sumac adventitious bud high frequency rate plant regeneration; This method is not subject to the restriction of season, time, and the plant offspring who obtains by the method makes a variation little, for the fast development of Chinese sumac eco-energy woods and molecular genetic improvement provide support.
Background technology
Chinese sumac is Anacardiaceae Rhus defoliation small arbor, it is a kind of important energy tree species of extensive distribution, also China Main Economic seeds, its seed oil content is up to 25%, unsaturated fatty acid content reaches 90%, is equivalent to the content in oil tea, and wherein linoleic acid content is at 70.4%-75.3%, far above the content in oil tea, there is very high health care as table oil and be worth.Chinese sumac has very high medical value simultaneously, root, stem, Ye Junke are used as medicine, and the formed gall nut of aphid insect gall colonized on its blade is important traditional Chinese medicine and the raw material of industry, be widely used in the industries such as medicine, chemical industry, printing and dyeing metallurgy, be China's important woods special product product, become the emphasis of countries in the world research.Along with the continuous increase of gall nut demand, the Chinese sumac kind of enlarging Chinese sumac woods, raising Gallnut yield, quality, cultivation high yield and high quality has become a kind of inexorable trend.
Sowing is transplanted and grown seedlings is the main seedling-cultivating method of current Chinese sumac, due to Rhus deep dormancy type, must carry out the husky Tibetan of low temperature and dewaxing treatment etc., after emerging, also needs to transplant, and process is loaded down with trivial details, waste time and energy, and sowing is subject to the restriction of time; Moreover seed is the position of squeezing edible oil and extracting various secondary metabolitess, utilize that seed sprouting not only wastes, reproductive efficiency is lower, and can't carry out Fast-propagation and popularization to some elite germplasms, far can not meet the fast development to demand and the Chinese sumac industry of seedling; In addition, due to the regenerating system of neither one efficient stable, make the research of Chinese sumac aspect the molecular genetic improvement in blank out.Therefore, be badly in need of the method for a kind of Chinese sumac highly efficient regeneration of exploitation, to meet the needs of current Chinese sumac seedling Fast-propagation and breed improvement.There is not yet up to now the report that relates to the Chinese sumac cultured in vitro is arranged both at home and abroad, more there are no the report of the high-frequency Chinese sumac plant regeneration technique by adventitious bud inducing.
Summary of the invention
For addressing the above problem, the present invention uses animal nutrition, and a kind of method of Chinese sumac adventitious bud high frequency rate plant regeneration is provided, thereby sets up a kind of efficient stable, Chinese sumac Multiple Buds renovation process fast.
The present invention adopts following technical scheme to achieve these goals:
A kind of method of Chinese sumac adventitious bud high frequency rate plant regeneration is characterized in that: at first get ripe Chinese sumac seed and be inoculated into after surface sterilization in the seed germination medium, through cultivating in 1-2 week, grow complete Chinese sumac plant; Under aseptic condition, obtain Chinese sumac cotyledon, cotyledonary node and hypocotyl sections, they are inoculated in respectively in cotyledon, cotyledonary node and hypocotyl adventitious bud induction culture base, obtain peak green indefinite bud clump after 25-30d; Peak green indefinite bud clump is moved in corresponding inducing culture and carries out the subculture cultivation, after growth 15-20d, grow the healthy and strong seedling of launching 2-4 sheet true leaf fully that 2-3cm is high; Seedling is transferred again and carry out culture of rootage in root media, 25-30d grows up to the healthy and strong test tube plantlet of well developed root system, after the hardening of finally being uncapped, transplants, and obtains healthy and strong Chinese sumac regrowth.
Described surface sterilization refers to: seed is processed after 30min the ethanol surface sterilization 1min with 75% with 98% the concentrated sulfuric acid, with 0.1% the mercuric chloride 3min that sterilizes, finally uses aseptic water washing 5 times afterwards, during sterilization, ceaselessly shakes.
Described cotyledon adventitious bud induction culture base is: MS+0.5-8.0mg/L 6-BA or 0.1-3.0mg/L TDZ+0.0-1.0mg/L IBA+30g/L sucrose+7.5g/L agar; Described cotyledonary node adventitious bud induction culture base is: MS+0.05-3.0 mg/L TDZ+0.0-0.5mg/L IBA+30g/L sucrose+7.5g/L agar; Described hypocotyl adventitious bud induction culture base: MS+0.25-8.0mg/L 6-BA or 0.05-3.0 mg/L TDZ+0.0-1.0mg/L IBA+30g/L sucrose+7.5g/L agar.
Described root media is: 1/2MS+0.5-2.0mg/L IBA+30g/L sucrose+7.5g/L agar.
Beneficial effect of the present invention is:
1, it is loaded down with trivial details that seedling raising process is transplanted in the Chinese sumac sowing, waste time and energy and be subject to seasonal restrictions, can not meet the demand of Chinese sumac establishment of economic forest, seriously restrict renewable sources of energy industry and take the development of the medicinal industry that gall nut is representative, the method of at present relevant Chinese sumac Multiple Buds highly efficient regeneration there is not yet report, Chinese sumac Multiple Buds highly efficient regeneration technology will provide important technical support for Fast-propagation and the molecular genetic improvement of China's Main Economic seeds Chinese sumac, to break relevant this research, still belong to blank situation at present;
2, the present invention confirms that carrying out the fast numerous of Chinese sumac seedling can pass through seed seedling-raising method and root platen press, thereby has saved the time, has removed time restriction and reproductive efficiency improves greatly, can be fast provides sufficient seedling for the construction of Chinese sumac economic forest; Simultaneously, the molecular improvement that this efficient renovation process is Chinese sumac particularly transgenic technology provides good basis.
The accompanying drawing explanation
Fig. 1 is the whole seed seedling.
Fig. 2 is cotyledon, cotyledonary node and hypocotyl indefinite bud clump, and wherein (a) is cotyledon indefinite bud clump, is (b) cotyledonary node indefinite bud clump, is (c) hypocotyl indefinite bud clump.
Fig. 3 is the healthy and strong plant seedling that Multiple Buds is taken root and obtained, and wherein (a) takes root for Multiple Buds, is (b) healthy and strong plant seedling.
Embodiment
Below in conjunction with embodiment, the present invention is described in further details.
Embodiment
A kind of method concrete operations of Chinese sumac adventitious bud high frequency rate plant regeneration are as follows:
1, at first seed processes after 30min the ethanol surface sterilization 1min with 75% with 98% the concentrated sulfuric acid, with 0.1% the mercuric chloride 3min that sterilizes, finally uses aseptic water washing 5 times afterwards, during sterilization, ceaselessly shakes;
2, the seed disinfected is placed on germination medium and is grown, through the cultivation of 1-2 about week, form 2 cotyledons and launch fully, the long hypocotylar whole seed seedling (Fig. 1) in 3-5cm left and right; The cotyledon of seed seedling, cotyledonary node, hypocotyl are all as the explant material of evoking adventive bud in the present invention;
3, the aseptic Chinese sumac cotyledon, cotyledonary node and the hypocotyl that obtain are seeded in the adventitious bud induction culture base, obtain light green indefinite bud clump (Fig. 2) after 30d, and inductivity can reach 95%.Cotyledon evoking adventive bud medium is MS+0.5-8.0mg/L 6-BA or 0.1-3.0mg/L TDZ+0.0-1.0mg/L IBA+30g/L sucrose+7.5g/L agar; Cotyledonary node adventitious bud induction culture base: MS+0.05-3.0 mg/L TDZ+0.0-0.5mg/L IBA+30g/L sucrose+7.5g/ agar L; Hypocotyl adventitious bud induction culture base: MS+0.25-8.0mg/L 6-BA or 0.05-3.0mg/L TDZ+0.0-1.0mg/L IBA+30g/L sucrose+7.5g/L agar;
The healthy and strong seedling that 2-3cm is high that has that 4, will obtain proceeds in root media, incubation growth 25-30d under illumination, can obtain the regeneration plant seedling (Fig. 3) of well developed root system, root media is 1/2MS+0.5-2.0mg/L IBA+30g/L sucrose+7.5g/L agar; The acclimatization and transplants of finally uncapping, obtain healthy and strong regrowth.
Claims (2)
1. the method for a Chinese sumac adventitious bud high frequency rate plant regeneration, is characterized in that: at first get ripe Chinese sumac seed and be inoculated into after surface sterilization in the seed germination medium, through cultivating in 1-2 week, grow complete Chinese sumac plant; Under aseptic condition, obtain Chinese sumac cotyledon, cotyledonary node and hypocotyl sections, they are inoculated in respectively in cotyledon, cotyledonary node and hypocotyl adventitious bud induction culture base, obtain peak green indefinite bud clump after 25-30d; Peak green indefinite bud clump is moved in corresponding inducing culture and carries out the subculture cultivation, after growth 15-20d, grow the healthy and strong seedling of launching 2-4 sheet true leaf fully that 2-3cm is high; Seedling is transferred again and carry out culture of rootage in root media, 25-30d grows up to the healthy and strong test tube plantlet of well developed root system, after the hardening of finally being uncapped, transplants, and obtains healthy and strong Chinese sumac regrowth; Described cotyledon adventitious bud induction culture base is: MS+0.5-8.0mg/L 6-BA or 0.1-3.0mg/L TDZ+0.0-1.0mg/L IBA+30g/L sucrose+7.5g/L agar; Described cotyledonary node adventitious bud induction culture base is: MS+0.05-3.0 mg/L TDZ+0.0-0.5mg/L IBA+30g/L sucrose+7.5g/L agar; Described hypocotyl adventitious bud induction culture base: MS+0.25-8.0mg/L 6-BA or 0.05-3.0 mg/L TDZ+ 0.0-1.0mg/L IBA+30g/L sucrose+7.5g/L agar; Described root media is: 1/2MS+0.5-2.0mg/L IBA+30g/L sucrose+7.5g/L agar.
2. the method for Chinese sumac adventitious bud high frequency rate plant regeneration according to claim 1, it is characterized in that: described surface sterilization refers to: seed is processed after 30min the ethanol surface sterilization 1min with 75% with 98% the concentrated sulfuric acid, afterwards with 0.1% mercuric chloride sterilization 3min, finally use aseptic water washing 5 times, ceaselessly shake during sterilization.
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Citations (2)
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| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| EP2311970A1 (en) * | 2008-04-28 | 2011-04-20 | Naturally Scientific Technologies Limited | Method for the production of bioproducts |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| EP2311970A1 (en) * | 2008-04-28 | 2011-04-20 | Naturally Scientific Technologies Limited | Method for the production of bioproducts |
Non-Patent Citations (6)
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| Abbas SAFARNEJAD,et al..Tissue culture in medicinal of sumac(Rhus coriaria).《L.J.S.N.》.2011,第2卷(第4期),参见对比文件1第760页右栏第2段、表1，第761页左栏第1-2段、表4. |
| Benamar benmahioul, et al..In vitro embryo germination and proliferation of pistachio (Pistacia vera L.).《Scientia Horticulturae》.2009,第122卷第479页右栏第2段，第480页左栏第8-24行，右栏第2段第1-2行. |
| In vitro embryo germination and proliferation of pistachio (Pistacia vera L.);Benamar benmahioul, et al.;《Scientia Horticulturae》;20091231;第122卷;第479页右栏第2段，第480页左栏第8-24行，右栏第2段第1-2行 * |
| Tissue culture in medicinal of sumac(Rhus coriaria);Abbas SAFARNEJAD,et al.;《L.J.S.N.》;20111231;第2卷(第4期);第760页右栏第2段、表1，第761页左栏第1-2段、表4 * |
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| 黄家德等.盐肤木种子繁殖试验.《广西植物》.1994,第14卷(第1期),第87页倒数第1段. |
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Effective date of registration: 20220127 Address after: 461143 group 2, Xingzhuang village, he Jie Xiang, Jian'an District, Xuchang City, Henan Province Patentee after: Henan flora Agricultural Technology Co.,Ltd. Address before: 230031 No. 350 Lake Road, Hefei, Anhui, Shushan Patentee before: HEFEI INSTITUTES OF PHYSICAL SCIENCE, CHINESE ACADEMY OF SCIENCES |