CN101057557A - Non-tube quick reproduction method for seedings of jatropha curcas L. - Google Patents
Non-tube quick reproduction method for seedings of jatropha curcas L. Download PDFInfo
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Abstract
The invention discloses a non-tube method for fast breeding aleurite seedling, comprising: taking disinfected stem carrying axillary bud of 2-3cm length as explant; enclosing its morphological upper notch, immersing its morphological lower notch in indolebutyric acid solution, cutting it in sand disk, dressing nutritional solution to keep sand disk wet, culturing it for 16. 2-20. 2 days under conditions of 25-28 Deg. C, 70-80% of relative humidity, 2000-3000 lx lighting strength and light period of 12 hour lighting/ 12 hour dark, and getting rooting and integral plant. The invention is characterized by fast breeding, free from influence of national factors such as season and region, low variation rate, simple process and low cost.
Description
Technical field
The present invention relates to the micropropagation of plants field, specifically, relate to the method for the little tung oil tree seedling of a kind of non-test tube rapid propagation.
Background technology
Little tung oil tree (Jatropha curcas L.), have another name called Jatropha curcas, the little seeds of a tung oil tree, pencil tree, the lotus tree, the cream paulownia, black soap, bright paulownia, the tar paulownia, blue or green paulownia wood, yellow swollen tree, sealing wax, tung tree, artificial flower is given birth to tree, the fiber crops baking is rare etc., be Euphorbiaceae (Euphorbiaceae) leprosy Pterostyrax (Jatropha) machaka or dungarunga, originate in America, now be distributed widely in the torrid zone, subtropics and dry-hot valley area, mainly be distributed in Guangdong in China, Hainan, Fujian, Taiwan, Guangxi, Yunnan, Guizhou, ground such as Sichuan (Chinese Plants will editorial board. Chinese Plants will [M]. Beijing: Science Press, 147-148.).Little tung oil tree has very high economic worth.Its complete stool can be used as medicine, nature and flavor are puckery, be slightly cold, poisonous (Kong Lingyi, Min Zhida, history swordsman etc. the chemical constitution study of leprosy tree root [J]. Botany Gazette, 1996,38 (2): 161-166.), subside a swelling among the people often being used to, analgesia, hemostasis, sterilization, desinsection, antipruritic, eliminate the wart body, emetic having loose bowels, anesthesia, diuresis, promotion wound healing and terminal pregnancy etc. (Liu Yonghong. the value of the little seeds of a tung oil tree and culture technique [J]. economic forest research, 2006,24 (4): 74-76.), little tung oil tree stem-leaf extract also is used to prevention and elimination of disease and pests (Li Jing, Wu Fenhong, Chen Yanyan etc. the jatropha curcas seed extract is to the insecticidal activity [J] of several insects. agricultural chemicals, 2006,45 (1): 57-58,65.).Meanwhile, little tung oil tree seed oil content up to 40～60% (Openshaw K.A review of Jatropha curcas:Anoil plant of unfulfilled promise[J] .Biomass and Bioenergy, 2000,19 (1): 1-15.), and seed oil is processed the substitute that just can be used as diesel oil slightly, it is the energy-source plant of desirable production biodiesel, receive much concern today deficient day by day at crude oil, the United Nations is widely used in the subtropics with little tung oil tree, the ecological construction of dry-hot valley poverty-stricken area, as the major project of the poverty alleviation (He Wenshu that helped and succeed, Xiao Ronggui, Yang Xiaoqiong etc. the DEVELOPMENT PROSPECT [J] of Jatropha curcas in poverty-stricken area development and ecological construction. Chinese traditional Chinese medicine information magazine, 2002,9 (10): 33-36.).In " about the implementation guideline of development bioenergy and biochemical industry property tax support policy " (wealth is built [2006] No. 702) of China's issue in 2006, clearly propose to encourage to utilize woody oil tree species such as little tung oil tree, Chinese pistache to be raw material processing bioenergy.The reply of National Development and Reform Committee's announcement subsequently implements to have in 24 living beings engineering hi-tech industrialization special projects in 2006 2 special supports to set up little tung oil tree implant mass base.
At present, the breeding and culturing of little tung oil tree mainly adopt traditional seminal propagation type of seeding and cottage propagation seedling-cultivating 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) is slow by the speed of self-sow, the little tung oil tree of seminal propagation, efficient is low.Little tung oil tree generally needs 1-3 from being seeded into ripe result, just enters the phase of abounding with on the 5th year, and a small amount of little tung oil tree of domestic present plantation is difficult to provide in a short time sufficient seed to be used for the implant mass construction of base.(2) the seminal propagation type of seeding is unfavorable for the maintenance of merit.(3) the cottage propagation seedling-cultivating method needs the little tung oil tree branch about 20～40cm length, diameter 2cm to make breeding unit materials (Yuan Lichun, Xu Zhongzhi, Wu Kui etc. Jatropha curcas planting technology [J]. the Yunnan agricultural science and technology, 2006, (1): 58.), little tung oil tree belongs to shrub or dungarunga, and the germplasm materials that a small amount of little tung oil tree of domestic present plantation is difficult to provide enough equally satisfies the needs of building implant mass base and afforestation.(4) planting seed and cuttage and seedling culture method are subjected to the influence of natural causes such as season, region big, and its operation has strict space and time requirement.
In order to solve the problem that traditional process for breeding and culturing exists, there is the researcher to carry out the research of test-tube plantlet method for quickly breeding, make explant with stem section, plumule, epicotyl, hypocotyl, cotyledon, endosperm, blade, petiole etc. respectively, successfully regeneration induction little tung oil tree whole plant (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.; Wei Qin, Lu Weida, Liao Yi etc. from Jatropha curcas epicotyl explant regeneration plant (English) [J]. Mol.Biol., 2004,30 (4): 475-478.; Chen Jinhong, Gao Min, Huang Jisheng. Jatropha curcas stem section cultured in vitro and breeding research [J] fast. Guangxi agricultural science, 2006,37 (3): 221-223.; The marquis wears, Zhang Shuwen, Yang Lin etc. Jatropha curcas endosperm callus induction and abatement of pollution thereof [J]. use and the environmental organism journal 2006,12 (2): 264-268.; Leeization, Ceng Ni, Jia Yongjiong etc. the fast numerous and root induction [J] of the short axillalry bud branch of Jatropha curcas. Sichuan University's journal (natural science edition), 2006,43 (5): 1116-1120.; Qin Hong, Song Songquan, Long Chunlin etc. the tissue culture of the little seeds of a tung oil tree and plant regeneration [J]. Yunnan plant research, 2006,28 (6): 649-652.).Though the test-tube plantlet quick-breeding method is than traditional process for breeding and culturing, improved individual reproduction speed, avoided individual reproduction to be subjected to the influence and the restriction of natural causes such as season, region, but complicated operation is loaded down with trivial details, plant needs strict sterile growing environment, the production cost height, and with test-tube seedling transplanting during to soil survival rate lower.In addition, in the tissue culture procedures, plant growth regulator such as test-tube plantlet Long contact time high concentration of cytokinin, archusia easily morph, and cause the forfeiture of part merit.Therefore the test-tube plantlet method for quickly breeding also is difficult to satisfy the demand of large-scale afforestation engineering to little tung oil tree sapling quality and quantity.
The non-tube rapid propagation technology of plant (The technique of efficient and rapid non-tubeplant cloning; TERNPC) (Yan Zhigang, Meng Aidong, Liu Yuan. the utilization [J] of the non-tube rapid propagation technology of plant on medicinal plant. Guangxi medical science, 2006,28 (6): 944-946.) be a new technology that on traditional cuttage raise seedling technique and tissue culture technique basis, grows up, be in manually operated illumination, temperature, under the damp condition, in conjunction with auxin, allow the root original hase of plant form fast, the technology that root system reaches full growth, it can make plant quantity realize multiplication by stages how much in than short-term, and this technology is produced significant for quick popularization and the industrialization of economically valuable plant.
The non-tube rapid propagation technology of plant if can be applied to the breeding of little tung oil tree seedling, that will be significant.
Summary of the invention
The objective of the invention is to overcome the problem that prior art exists, provide a kind of reproduction speed fast, not influenced by natural causes such as season, region, aberration rate is low, and step is simple, the method for the little tung oil tree seedling of the non-test tube rapid propagation that cost is low.
To achieve these goals, the method for the little tung oil tree seedling of non-test tube rapid propagation of the present invention adopts following steps: make explant with the length 2～3cm after the sterilization, the little stem section of little tung oil tree of carrying axillalry bud; Seal its morphology upper end otch, after its morphology lower end otch is soaked in indolebutyric acid (IBA) solution, cuttage is in sand table, imposing nutrient solution keeps sand table moistening, at 25～28 ℃ of temperature, relative moisture 70～80%, intensity of illumination 2000～3000lx, cultivated 16.2～20.2 days under the condition of photoperiod 12h illumination/12h dark, can take root, the regeneration whole plant.
In said method, the described preferred 1-2 of little stem section that carries axillalry bud gives birth to little tung oil tree branch.
In said method, and the preferred 1/2MS minimal medium of described nutrient solution (Gui Yaolin, horse is sincere. Plant Tissue Breeding [M]. and Beijing: Science Press, 1985, p23-24), pH 5.8～6.0, and it contains the Murashige﹠amp that concentration reduces by half; Skoog medium macroelement, trace element and molysite composition.
In said method, the preferred 0.2～2.0mg/L of the concentration of described indolebutyric acid solution.
In said method, described sealing preferably seals with the solid paraffin of fusing.
In said method, preferred 1 hour of the time of described indolebutyric acid solution soaking explant.
Compared with prior art, the present invention has following beneficial effect:
The used explant of the present invention be 2～3cm, the little stem section of little tung oil tree of carrying axillalry bud, compare with the cuttage branch of the long 20～40cm of tradition, used explant quantity reduces about 7～20 times, has greatly saved germplasm materials, has improved fast numerous efficient.
2. the used explant of the present invention intercepts from childhood tung oil tree 1-2 and gives birth to branch, though this kind branch lignification, degree of lignification deeply, does not still keep the vigorous growth state, thereby the cuttage fringe easily takes root, and not perishable.
3. the used explant of the present invention intercepts 1～2 year living branch of tung oil tree from childhood, and cultivating little tung oil tree is shrub, needs constantly in time to prune branch, promote collateral development, keep suitable plant height and shape of tree-crown, to increase output and to be convenient to picking fruit, thereby the explant material amount is big, and is easy to obtain.
4. the present invention adopts 1mg/L IBA solution soaking cuttage fringe 1h, 18.2 just can obtain complete regenerated plant in ± 2.0 days, it is fast not only to obtain individual speed than traditional process for breeding and culturing, and faster than the speed of test-tube plantlet method for quickly breeding acquisition complete regenerated plant in the existing report.
5. the present invention is at in-house operation, thereby is not subjected to the influence and the restriction of natural causes such as season, region, can breed whenever and wherever possible.
6. the invention belongs to cottage propagation technology category, the branch that is produced by axillary bud growth keeps former clonal genotype and phenotype, has avoided the natural seed breeding to be unfavorable for the maintenance of merit and the individual variation rate height that the test-tube plantlet quick-breeding method obtains, the defective that may lose the part merit in tissue culture procedures.
The present invention adopted with little stem segment cuttage in sand table, directly take root, forming seedling through one step culture, thereby avoided test-tube plantlet root leaf development in blake bottle not exclusively and with test-tube plantlet from medium, to transplant the low defective of survival rate to the soil course.
8. the present invention only carries out sterilization treatment to little stem section, thereby compare the test-tube plantlet method for quickly breeding, need not strict sterile tissue and cultivate operation, less demanding to fast numerous equipment, it is low not only to produce regeneration seedling cost, and method of operating is simple, is easy to grasp, be applicable to the factorial seedling growth of little tung oil tree, thereby for afforestation with build extensive little tung oil tree planting base good sapling is provided.
Description of drawings
Fig. 1 is that little stem segment cuttage is in sand table;
Fig. 2 goes out young leaves for little stem segment length;
Fig. 3 is transplanted into the seedbed for seedling.
Embodiment
Embodiment and comparative example all adopt 1 year little tung oil tree branch of life to intercept little stem section and breed fast.
Sterilization treatment: will adopt the branch defoliation that comes, and be soaked in 20min in the thimerosal [5% clorox (NaClO), 0.1%Tween-20], then with running water flushing 5 times; Branch is cut into shorter stem section about 15～20cm, in 70% ethanol, soaks 10min, in thimerosal, soak 15min then, use aseptic water washing 5 times, with the remaining clorox of thorough flush away.
Preparation archusia indolebutyric acid (IBA) solution; Concentration is respectively 0.2,0.5,1.0,2.0 (mg/L).
The configuration nutrient solution: preparation 1/2MS minimal medium, pH 5.8～6.0, and concrete prescription sees Table 1.
Table 1
| Composition | Molecular weight | Working concentration (mg/L) | |
| Macroelement | KNO 3 NH 4NO 3 KH 2PO 4 MgSO 4.7H 2O CaCl 2.2H 2O | 101.11 80.04 136.09 246.47 147.02 | 950 825 85 185 220 |
| Trace element | KI H 3BO 3 MnSO 4.4H 2O ZnSO 4.7H 2O | 166.01 61.83 223.01 287.54 | 0.42 4.0 11.2 5.0 |
| Na 2MoO 4.2H 2O CuSO 4.5H 2O CoCl 2.6H 2O | 241.95 249.68 237.93 | 0.13 0.01 0.01 | |
| Molysite | Na 2.EDTA FeSO 4.7H 2O | 372.25 278.03 | 18.7 13.9 |
Non-test tube rapid propagation: the stem section after will sterilizing is cut into the long little stem section of 2～3cm under aseptic condition, be with an axillalry bud on each little stem section, dries on aseptic super-clean bench; The morphology upper end otch of little stem section is dipped the solid paraffin of fusing, take out cooled and solidified rapidly; The morphology lower end otch of little stem section is soaked in 1h in the solution that contains variable concentrations archusia IBA, and cuttage imposes nutrient solution and keeps the moistening (see figure 1) of sand table in sand table then.Keep the about 2000～3000lx of intensity of illumination, periodicity of illumination 12h illumination/12h dark, 25～28 ℃ of temperature, relative moisture 70～80%.From the 8th day, estimate the situation of taking root every day; The 28th day statistics calculated rooting rate, measures new root length, adopts duncan's new multiple range method to carry out significance of difference analysis.Wherein, total root length is the sum that adds up of all new root length on the little stem section.Statistics sees Table 2.
Treat that little stem Duan Xingen grows to 0.5～1.0cm, bears (see figure 2) behind the 1-2 sheet young leaves, sand table was placed outdoor shady and cool ventilation place hardening 2 days, the seedling of will regenerating then is transplanted into and is coated with (see figure 3) in the good seedbed of nutrition soil, water penetration, but field planting land for growing field crops when treating plant height 50～100cm.
Table 2
| Embodiment | IBA (mg/L) | Explant number (individual) | Rootage duration (my god) mean ± SD | Rooting rate (%) | New root quantity (bar) mean ± SD | Long (cm) mean ± SD of total root |
| 1 2 | 0.2 0.5 | 30 30 | 21.8±1.8 c 21.7±1.8 c | 63.3 b 73.3 b | 5.3±1.3 b 5.4±1.3 b | 4.3±1.9 b 4.5±2.5 b |
| 34 comparative examples 1 | 1.0 2.0 0.0 | 30 30 30 | 18.2±2.0 a 20.3±2.5 b 23.6±2.1 d | 96.7 a 80.0 b 56.7 b | 6.3±1.8 a 4.4±1.9 b 4.2±1.6 c | 6.8±3.5 a 4.3±3.0 b 2.7±2.1 c |
The result shows: IBA soaks the acquisition that helps non-tube rapid propagation seedling, compares embodiment 1～4 and comparative example 1, and embodiment 1-4 is under the IBA soaking conditions, the rooting rate of the little stem section of little tung oil tree is than higher, take root than comparatively fast, root system is relatively more flourishing, transplants the back growth rapidly.Wherein, the most pronounced effects of embodiment 3, embodiment 3 adopts 1mg/L IBA to soak the little stem section of little tung oil tree 1h, and rooting rate reaches 96.7%, and rootage duration is 18.2 ± 2.0 days, and the quantity of new root is 6.3 ± 1.8, total root reaches 6.8 ± 3.5cm.
Claims (6)
1. the method for the little tung oil tree seedling of non-test tube rapid propagation is characterized in that comprising the steps: making explant with the length 2～3cm after the sterilization, the little stem section of little tung oil tree of carrying axillalry bud; Seal its morphology upper end otch, its morphology lower end otch is soaked in the indolebutyric acid solution; Immersion finishes the back cuttage in sand table, imposes nutrient solution and keeps sand table moistening, 25～28 ℃ of temperature, relative moisture 70～80%, intensity of illumination 2000～3000lx cultivated 16.2～20.2 days under the condition of every day illumination half a day and dark half a day, can take root, the regeneration whole plant.
2. the method for the little tung oil tree seedling of non-test tube rapid propagation according to claim 1 is characterized in that described little stem section intercepts 1～2 year living branch of tung oil tree from childhood.
3. the method for the little tung oil tree seedling of non-test tube rapid propagation according to claim 1, the concentration that it is characterized in that described indolebutyric acid solution is 0.2～2.0mg/L.
4. the method for the little tung oil tree seedling of non-test tube rapid propagation according to claim 1 is characterized in that described nutrient solution is the 1/2MS minimal medium, and pH 5.8～6.0.
5. the method for the little tung oil tree seedling of non-test tube rapid propagation according to claim 1 is characterized in that described sealing is the solid paraffin sealing with fusing.
6. the method for the little tung oil tree seedling of non-test tube rapid propagation according to claim 1, the time that it is characterized in that described indolebutyric acid solution soaking explant is 1 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710028385A CN101057557B (en) | 2007-06-01 | 2007-06-01 | Non-tube quick reproduction method for seedings of jatropha curcas L. |
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| CN102379197A (en) * | 2010-08-25 | 2012-03-21 | 伍玉雄 | Aleurites montana large-area seedling raising method |
| CN102696480A (en) * | 2012-05-07 | 2012-10-03 | 广西壮族自治区林业科学研究院 | Method for culturing aseptic oil tea buds in test tube by using sand matrix |
| CN103460927A (en) * | 2013-09-04 | 2013-12-25 | 深圳星宇投资控股有限公司 | Intermediate propagation method for jatropha curcas |
| CN101952447B (en) * | 2008-02-01 | 2014-04-02 | 淡马锡生命科学研究院有限公司 | Regeneration and mass propagation of jatropha curcas through somatic embryogenesis |
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