CN103355168B - Ficus tikoua rapid propagation method - Google Patents
Ficus tikoua rapid propagation method Download PDFInfo
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- CN103355168B CN103355168B CN201310302401.8A CN201310302401A CN103355168B CN 103355168 B CN103355168 B CN 103355168B CN 201310302401 A CN201310302401 A CN 201310302401A CN 103355168 B CN103355168 B CN 103355168B
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Abstract
The invention discloses a ficus tikoua rapid propagation method. According to the method, ficus tikoua terminal buds or axillary buds with stem sections are cultured in an initial medium, such that aseptic seedlings are obtained; clustered buds are induced on a proliferation medium; and rooting is carried out on a rooting medium. When ficus tikoua tissue culture is carried out with the set of technical methods provided by the invention, the period is short, propagation coefficient is high, cost is low, and high factory production capacity is provided. When propagation is carried out with the method provided by the invention, ficus tikoua can be effectively protected, developed, and utilized.
Description
Technical field
The invention belongs to the quick propagating technology field of ground loquat, particularly the method for the tissue-culturing quick-propagation of a kind of ground loquat (Ficus tikoua).
Background technology
Ground loquat (Ficus tikoua) belongs to Moraceae Ficus, claims again pachyrhizus, digua fig stem and leaf, ground courage purple, Fructus Fici tikouae.The perennial fallen leaves shrub that crawls, complete stool has milk, originates in the ground such as Guangxi, Guizhou, Hunan, in Guizhou and the western Hunan as Miao ethnic group's conventional crude drugs.Both at home and abroad the pharmaceutical research of this platymiscium is shown, that this platymiscium has is hypoglycemic, relaxing smooth muscle, the effect such as antitumor, antibacterial.It is edible, delicious fragrant and sweet that ground loquat also can make fruit, lovely luster, and its fruit is as shown in Figure 1.Ground loquat also can do to see fruit, see leaf hanging potted landscape, and pliable and tough emerald green stem is climing graceful elegant, has the beautiful scenery of " green silk silk ribbon ten thousand hangs down ".Ground loquat is also suitable for park or subjects to the place greening that visitor tramples on, and can bear numerous visitors and trample on.Ground loquat is always growing because this tough and tensile and in great numbers stem is climing crisscrossly, will naturally " knit " like this one " huge fishing net ", can be loosened the soil in surface, sandy soil " catch " firmly, add dense leaf " front cover " all the year round, exactly like and added a cover a permanent green carpet, the solid native special efficacy of this sand prevention be other ground cover plant institute can not and, can be rated as sand prevention and consolidate native nonsuch.In sum, ground loquat is to integrate multi-purpose plant edible, medicinal, that afforest, view and admire.
Ground loquat artificial propagation mainly adopts cottage propagation, but wild resource is very limited, and conventional propagation technique cannot provide quantity huge seedling in a short period of time, is difficult to commercialize.Only have the artificial Fast-propagation of solution, expand the planting number of ground loquat, the planting base of setting up commercial development utilization just can make this precious resources obtain the most effectively protecting and developing, and realizes the real valency of its resource and grows.It is investigated inquiry, there is no the report of ground loquat Study on tissue culture and patent application at present both at home and abroad.
Summary of the invention
The object of the invention is to, provide a kind of ground loquat (Ficus tikoua), the method for tissue-culturing quick-propagation; The ground loquat Multiple Buds that the method energy rapid induction is a large amount of, and growth coefficient and rooting rate high, easy to operate, there is good application prospect.
The object of the invention is to realize in the following manner.
A method for quickly breeding for ground loquat, comprises the following steps:
Getting the long terminal bud with stem section of ground loquat (Ficus tikoua) 2～3cm or axillalry bud is inoculated into just in culture base MS+6-BA0.5-1.5mg/L+IAA0.1-0.8mg/L and cultivates and obtain aseptic seedling, then aseptic seedling is carried out to shoot proliferation and culture of rootage, finally carry out hardening and transplanting.
In said method, in first culture base, the concentration of 6-BA is preferably 0.8mg/L; The concentration of IAA is preferably 0.3mg/L.
In said method, shoot proliferation medium used is: molysite+6-BA1.0-2.5mg/L+KT0.2-0.5mg/L+IAA0.1-0.8mg/L+ paclobutrazol 0.5-2.0mg/L of organic principle+MS medium of trace element+MS medium of macroelement+MS medium of N6 medium.
In said method, in subculture proliferated culture medium, the concentration of 6-BA is preferably 1.0mg/L; The concentration of KT is preferably 0.5mg/L; The concentration of IAA is preferably 0.2mg/L; The concentration of paclobutrazol is preferably 1.0mg/L.
In said method, culture of rootage medium used is: MS+IAA0.1-0.8mg/L+NAA0.1-0.8mg/L+KT0.5-1.0mg/L+ paclobutrazol 0.2-1.0mg/L.
In said method, in root media, the concentration of IAA is preferably 0.2mg/L; The concentration of NAA is preferably 0.5mg/L; The concentration of KT is preferably 0.5mg/L; The concentration of paclobutrazol is preferably 0.5mg/L.
Test-tube plantlet hardening and transplanting process in said method: by the test-tube plantlet of taking root, natural daylight lower refining seedling 5～7 days, wash root medium off, be transplanted in the matrix of sand, perlite, the mixing of peat soil equal-volume, spraying keeps humidity in air to reach 85%～95%, 20～30 DEG C of cultivations, transplant land for growing field crops for 30～40 days.
In said method, just before culture, the terminal bud with stem section or axillalry bud are sterilized, detailed process is:
Get terminal bud or the axillalry bud of ground loquat with stem section, first clean up with washing powder, on aseptic operating platform, with aseptic washing 5～7 times, with 70% alcohol infiltration 1 minute, then put into 0.1% mercuric chloride solution sterilizing 12～18 minutes, aseptic water washing 5～6 times.
Condition of culture in said method is at 24 ± 2 DEG C, every day 10 ± 2h) illumination, intensity of illumination is 1500-2000lx.
In said method, the terminal bud with stem section of ground loquat or axillalry bud are cultivated after 4 weeks in first culture base, explant all grows 2-5 joint, is highly the aseptic seedling of 3-6cm; Then aseptic seedling is cut into stem section, each stem section is with a terminal bud or an axillalry bud; Stem section is seeded in shoot proliferation medium and is cultivated, cultivate after 4 weeks, each stem section all grows again the aseptic seedling of 2-6 joint, goes out more than 3 Multiple Buds from stem segment base minister simultaneously; First culture or shoot proliferation are cultivated to the aseptic seedling obtaining to be cut into be inoculated in root media, to cultivate after 4 weeks with the stem section of bud and to go out 2-4 bar root from stem segment base minister.
Taking ground loquat, the terminal bud with stem section or axillalry bud are explant in the present invention, utilize medium of the present invention, have opened up a kind of Fast-propagation ground loquat aseptic seedling and have kept the new method of its mother plant hereditary capacity.The inventive method has the following advantages and beneficial effect:
1) this method has overcome conventional method and has bred slow defect, in a short period of time can the needed seedling of large-scale breeding; This method is drawn materials easily, and the process that obtains aseptic seedling is simple, and reproduction coefficient is high, and within 4 weeks, propagation multiple can reach more than 10, and group training seedling rooting rate reaches more than 80%, and transplanting survival rate can reach more than 90%;
2) this method production cost is low, and using value is high, emerges fast, stable high, seedling quality better, and resistance, has very strong batch production production capacity;
3) utilize method of the present invention breeding spot loquat group training seedling, can more effectively protect and develop ground loquat.
Brief description of the drawings
Fig. 1 is the photo of the edible fruit of ground loquat;
Fig. 2 is the just terminal bud of culture and the photo of axillary bud growth situation of the present invention ground loquat;
Fig. 3 be the present invention ground loquat shoot proliferation cultivate 1 generation Multiple Buds growing state photo;
Fig. 4 be the present invention ground loquat shoot proliferation cultivate 5 generation Multiple Buds growing state photo;
Fig. 5 is the photo of the present invention ground loquat rooting of vitro seedling situation;
Fig. 6 is the photo that the present invention ground loquat test-tube seedling transplanting survives situation.
Embodiment
Be intended to further illustrate the present invention below in conjunction with embodiment, and unrestricted the present invention.
Embodiment 1, tissue-culturing quick-propagation ground loquat aseptic seedling
One, medium preparation and sterilizing
Just culture base: MS+6-BA0.8mg/L+IAA0.3mg/L;
Subculture medium: N6 macroelement+MS trace element+MS organic principle+MS molysite+6-BA1.0mg/L+KT0.5mg/L+IAA0.2mg/L+ paclobutrazol (CCC) 1.0mg/L;
Root media: MS+IAA0.2mg/L+NAA0.5mg/L+KT0.5mg/L+ paclobutrazol (CCC) 0.5mg/L;
At 121 DEG C of above medium, sterilizing 20 minutes.
Two, first culture
Get terminal bud or the axillalry bud (the overall length 2-3cm of stem section and bud) of ground loquat (Ficus tikoua) with stem section, first clean up with washing powder, on aseptic operating platform with aseptic washing 5～7 times, with 70% alcohol infiltration 1 minute, put into again 0.1% mercuric chloride solution sterilizing 12～18 minutes, aseptic water washing 5～6 times, terminal bud with stem section or axillalry bud are proceeded to and be equipped with just on culture base in triangle bottle (50 milliliters of capacity), each triangular flask explant number is 3, under following condition of culture, cultivate: 24 DEG C of left and right, illumination 10 hours every days, intensity of illumination is 1500-2000lx,
9 of terminal bud inoculations, 21 of axillalry bud inoculations.Cultivate 4 weeks, these 30 terminal buds and axillalry bud all grow 2-5 joint aseptic seedling, and aseptic seedling height is 2-6cm, and each strain group training seedling is a strain.Terminal bud and axillary bud growth situation are as shown in Figure 2.
Three, shoot proliferation is cultivated
20 strains of getting in above-mentioned 30 strains are carried out successive propagation, and concrete grammar is as follows: the aseptic seedling that grows 2-5 joint is cut into stem section, and each stem section is with a terminal bud or an axillalry bud.Above-mentioned stem section is seeded in subculture medium and is cultivated, the same step 2 of condition of culture;
Above-mentioned stem section, in proliferated culture medium, cultivates after 4 weeks, and each stem Duan Jun grows the aseptic seedling of 2-6 joint, goes out more than 3 Multiple Buds from stem segment base minister simultaneously.Multiple Buds growing state as shown in Figure 3;
This aseptic seedling more according to the method described above every 4 weeks subcultures once, subculture 5 times.Subculture 1 time, each stem Duan Jun grows the aseptic seedling of 2-6 joint, and aseptic seedling height is 3-6cm, goes out 4-6 Multiple Buds from stem segment base minister simultaneously, and reproduction coefficient reaches more than 10.Multiple Buds growing state as shown in Figure 4.
Four, culture of rootage
By above-mentioned aseptic seedling as stated above, be cut into stem section, be transferred on the root media of N6+IAA0.3mg/L+NAA0.5mg/L+KT-1.0mg/L+ paclobutrazol (CCC) 0.5 the same step 2 of condition of culture; Cultivate 4 weeks, each stem Duan Jun grows the aseptic seedling of 2-6 joint, and aseptic seedling height is 3-6cm, goes out 2-4 bar root from stem segment base minister, and rooting rate reaches more than 85%.Take root seedling growing state as shown in Figure 5.
Five, acclimatization and transplants
In the time that test-tube plantlet stem segment base minister goes out 2-4 bar root, natural daylight lower refining seedling 5～7 days, wash root medium off, be transplanted in the matrix of sand, perlite, the mixing of peat soil equal-volume, spraying and moisturizing 85%～95%, is incubated 20～30 DEG C of cultivations, within 30～40 days, transplant land for growing field crops, survival rate reaches more than 90%.Test-tube seedling transplanting survives situation as shown in Figure 6.
Ground provided by the present invention loquat tissue culture propagation, the advantage such as the terminal bud with stem section or axillalry bud are explant, draw materials easily, and quantity is large, and genetic stability is high, well-grown.In incubation, not only terminal bud or axillary bud growth speed are fast, and stem segment base portion can synchronously induce Multiple Buds, and reproduction coefficient is large.Culture of rootage aseptic seedling can be cut into terminal bud equally or stem section proceeds in root media, and not only terminal bud or axillary bud growth speed are fast, and can synchronously induce root.
Embodiment 2, tissue-culturing quick-propagation ground loquat aseptic seedling
Medium preparation and sterilizing
Just culture base: MS+6-BA1.0mg/L+IAA0.5mg/L;
Subculture medium: N6 is a large amount of+and MS trace+MS is organic+MS molysite+6-BA1.0mg/L+KT0.3mg/L+IAA0.2mg/L+ paclobutrazol (CCC) 0.5mg/L;
Root media: MS+IAA0.3mg/L+NAA0.3mg/L+KT0.5mg/L+ paclobutrazol (CCC) 0.5mg/L.
At 121 DEG C of above medium, sterilizing 20 minutes;
Get ground loquat (Ficus tikoua) terminal bud or stem section, basic operation method is consistent with embodiment 1, propagation, and rooting efficiency is worse than embodiment 1 slightly, but also can reach fine.
Embodiment 3, tissue-culturing quick-propagation ground loquat aseptic seedling
Just culture base: MS+6-BA0.5-1.5mg/L+IAA0.1-0.8mg/L
Shoot proliferation medium is: molysite+6-BA1.0-2.5mg/L+KT0.2-0.5mg/L+IAA0.1-0.8mg/L+ paclobutrazol 0.5-2.0mg/L of organic principle+MS medium of trace element+MS medium of macroelement+MS medium of N6 medium.
Culture of rootage medium is: MS+IAA0.1-0.8mg/L+NAA0.1-0.8mg/L+KT0.5-1.0mg/L+ paclobutrazol 0.2-1.0mg/L.
In the concentration range of above-mentioned several medium, carry out according to the operating procedure of embodiment 1, also substantially can reach more satisfied effect.
Claims (7)
1. a method for quickly breeding for ground loquat, is characterized in that, comprises the following steps:
Get the long terminal bud with stem section or the axillalry bud of ground loquat (Ficus tikoua) 2～3cm, be inoculated in first culture base MS+6-BA0.5-1.5mg/L+IAA0.1-0.8mg/L and cultivate and obtain aseptic seedling, then aseptic seedling is carried out to shoot proliferation and culture of rootage, finally carry out hardening and transplanting;
Shoot proliferation medium used is:
Molysite+6-BA1.0-2.5mg/L+KT0.2-0.5mg/L+IAA0.1-0.8mg/L+ paclobutrazol 0.5-2.0mg/L of organic principle+MS medium of trace element+MS medium of macroelement+MS medium of N6 medium;
Culture of rootage medium used is:
MS+IAA0.1-0.8mg/L+NAA0.1-0.8mg/L+KT0.5-1.0mg/L+ paclobutrazol 0.2-1.0mg/L;
Test-tube plantlet hardening and transplanting process:
By the test-tube plantlet of taking root, natural daylight lower refining seedling 5～7 days, wash root medium off, be transplanted in the matrix that sand, perlite, peat soil equal-volume mixes, spraying keeps humidity in air to reach 85%～95%, and land for growing field crops is transplanted in 20～30 DEG C of cultivations for 30～40 days.
2. the method for quickly breeding of loquat according to claim 1ly, is characterized in that: in first culture base, the concentration of 6-BA is 0.8mg/L; The concentration of IAA is 0.3mg/L.
3. the method for quickly breeding of loquat, is characterized in that according to claim 1ly, and in shoot proliferation medium, the concentration of 6-BA is 1.0mg/L; The concentration of KT is 0.5mg/L; The concentration of IAA is 0.2mg/L; The concentration of paclobutrazol is 1.0mg/L.
4. the method for quickly breeding of loquat, is characterized in that according to claim 1ly, and in root media, the concentration of IAA is 0.2mg/L; The concentration of NAA is 0.5mg/L; The concentration of KT is 0.5mg/L; The concentration of paclobutrazol is 0.5mg/L.
5. the method for quickly breeding of loquat according to claim 1ly, it is characterized in that: before first culture, the terminal bud with stem section or axillalry bud are sterilized, detailed process is: get terminal bud or the axillalry bud of ground loquat with stem section, first clean up with washing powder, on aseptic operating platform with aseptic washing 5～7 times, infiltrate 1 minute with 70% alcohol, then put into 0.1% mercuric chloride solution sterilizing 12～18 minutes, aseptic water washing 5～6 times.
6. according to the method for quickly breeding of the ground loquat described in any one in claim 1-5, it is characterized in that: the condition of culture in described method is at 24 ± 2 DEG C 10 ± 2h illumination every day, intensity of illumination 1500-2000lx.
7. according to the method for quickly breeding of the ground loquat described in claim 1-5 any one, it is characterized in that: the terminal bud with stem section of ground loquat or axillalry bud are cultivated after 4 weeks in first culture base, and explant all grows 2-5 joint, is highly the aseptic seedling of 3-6cm; Then aseptic seedling is cut into stem section, each stem section is with a terminal bud or an axillalry bud; Stem section is seeded in shoot proliferation medium and is cultivated, cultivate after 4 weeks, each stem section all grows again the aseptic seedling of 2-6 joint, goes out more than 3 Multiple Buds from stem segment base minister simultaneously; First culture or shoot proliferation are cultivated to the aseptic seedling obtaining to be cut into be inoculated in root media, to cultivate after 4 weeks with the stem section of bud and to go out 2-4 bar root from stem segment base minister.
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| CN103518625B (en) * | 2013-11-01 | 2015-11-25 | 重庆文理学院 | The tissue culture medium (TCM) of Root of Fiddleleaf Fig blade and in-vitro regeneration method |
| CN104620827B (en) * | 2015-02-11 | 2017-01-18 | 中南大学 | Cutting propagation method for ficus tikoua |
| CN105325296B (en) * | 2015-11-13 | 2017-10-24 | 广西壮族自治区药用植物园 | A kind of jackfruit quick breeding method for tissue culture |
| CN105325300B (en) * | 2015-11-30 | 2017-10-20 | 华南农业大学 | The methods and applications of Eriobotrya distant hybrid progeny material are quickly bred by embryo culture |
| CN107372107A (en) * | 2017-07-26 | 2017-11-24 | 兰溪市顺光园艺技术有限公司 | The fast culture process of ground loquat |
| CN110432055A (en) * | 2019-06-24 | 2019-11-12 | 铜仁学院 | A kind of Moraceae ficus species ground fruit cultural method |
| CN110432054A (en) * | 2019-06-24 | 2019-11-12 | 铜仁学院 | A kind of Moraceae ficus species ground long climing thick root seedling fostering method of fruit |
| CN112438152A (en) * | 2020-11-02 | 2021-03-05 | 铜仁学院 | Large-scale efficient breeding method for Diguo seedlings |
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| JP2002233360A (en) * | 2001-02-06 | 2002-08-20 | Univ Hiroshima | Cultured cell of ficus thunbergii maxim., and method for tissue culture using the same |
| JP3787624B2 (en) * | 2002-10-04 | 2006-06-21 | 独立行政法人農業・食品産業技術総合研究機構 | Method for inducing adventitious roots and adventitious buds in figs |
| CN102232360B (en) * | 2011-04-25 | 2012-12-12 | 安徽农业大学 | Method for establishing in-vitro rapid propagation system of anubias barteri var. barteri, anubias barteri var. nana, ficus henryi and anubias hastifolia |
| CN103053416A (en) * | 2012-12-26 | 2013-04-24 | 中国长江三峡集团公司 | Method for tissue culture and rapid propagation of ficus elastica |
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