CN103314848B - Rapidly breeding method for ampelocalamus luodianensis - Google Patents
Rapidly breeding method for ampelocalamus luodianensis Download PDFInfo
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Abstract
The invention discloses a rapidly breeding method for ampelocalamus luodianensis, comprising: explant selection of the ampelocalamus luodianensis, explant disinfection of the ampelocalamus luodianensis, multiple shoot induction of the ampelocalamus luodianensis, multiple shoot propagation of the ampelocalamus luodianensis, multiple shoot rhizogenesis of the ampelocalamus luodianensis, and tissue culture plant transplanting of the ampelocalamus luodianensis. The rapidly breeding method for ampelocalamus luodianensis has advantages of small investment, fast seedling growing speed of the ampelocalamus luodianensis, high survival rate and consistent growth vigor of the sprouts, and overcomes various disadvantages of traditional mother bamboo transplanting, such as low breeding efficiency, large damage of ecological environment, long breeding period and high transport cost. As the rapidly breeding method for ampelocalamus luodianensis, the breeding of ampelocalamus luodianensis by the method is not limited by seasons, transport cost is low, and the method is suitable for industrial seedling growing production, and provides a rapid and effective approach for rapidly recovering amount of the ampelocalamus luodianensis in a natural state and performing ecological benefits and economic benefits of the ampelocalamus luodianensis in karst areas.
Description
Technical field
The invention belongs to bamboo plant group culturation rapid propagating technology field, specifically relate to a kind of method that adopts tissue culture technique to set up the fast traditional font of drepanostachyum luodianense system
Background technology
Drepanostachyum luodianense (Ampelocalamus luodianensis) is outstanding Sinobambusa (Ampelocalamus) the bamboo kind of grass family Bambusoideae, for the endangered plants of karst, be mainly distributed at present Luodian County, Guizhou Province, Changshun County, Miao-Bouyei Autonomous County of Ziyun, Wangmo County.Whole area is positioned at north latitude: 24 ° 58 '-26 ° 03 '; East longitude: 105 ° 25 '-107 ° 03 ', vertical distribution height is between 650-1250m.Its good looking appearance, there is cultivation ornamental value, simultaneously, drepanostachyum luodianense can be at the severe karst well-grown of environment, can be drought-resistant and barren, solid soil, water conservation, fertilizer-preserving ability effect to enhancing soil are very remarkable, its forest canopy is obviously better than fruticeta to the interception of rainfall, in sylvan life soil, the quantity of water-stable aggregate is also higher than fruticeta and meadow, in Karst ecosystem, the ecological functions such as water source self-restraint, water and soil conservation, nutrient balance are played an important role, there are very high ecological benefits.In the last few years, drepanostachyum luodianense is widely used as paper making raw material by local papermaking enterprise, particularly serious to the young bamboo predation of new life formula felling phenomenon, cause drepanostachyum luodianense clonal population serious degradation, population quantity sharply reduces, and in " Chinese species Red List (first volume: Red List) ", drepanostachyum luodianense is listed in utmost point danger species.
Bamboo plant blooming cycle is long, be difficult to take the mode of sexual propagation to obtain bamboo seedling, at present the modes of reproduction of bamboo plant mainly adopts and moves bamboo, buries the modes of reproduction such as whip, cuttage and obtain bamboo seedling, and there are many problems in all class modess of reproduction, as: move a large amount of female bamboo of bamboo afforestation process need, and cost of transportation is high, survival rate is low, reproduction speed is slow; Bury in whip seedling raising process that bamboo seedling reproduction coefficient is low, growth rate slow and bamboo seedling later stage growing way is weak, the cycle of growing into forest is long; Though cuttage and seedling culture reproduction coefficient is high compared with first two mode, in branch cutting process, be subject to seasonal restrictions obviously and survival rate is low, growing-seedling period is long.The breeding and afforestation mode of drepanostachyum luodianense mainly adopts and moves bamboo afforestation at present, and efficiency is extremely low, cannot effectively alleviate the problem that the current population quantity of drepanostachyum luodianense sharply reduces.The method efficiency that adopts tissue to cultivate is high, condition is controlled, and bamboo seedling growth is fast, the cycle is short, reproduction coefficient is high and be not subject to seasonal restrictions, convenient transportation, the batch production that is applicable to drepanostachyum luodianense bamboo seedling is produced, and can fundamentally alleviate the problem that drepanostachyum luodianense population quantity declines.
Up to now, Bamboo Tissue training both at home and abroad makes great progress, by tissue culture technique, realize the tissue-culturing rapid propagation to Ornamental Bamboo such as safety bamboo (Pseudosasa japonica ' Tsutsumiana'), black bamboo (Phyllostachys nigra), gold inlaid jade bamboo (Phyllostachys aureosulcata ' Spectabilis'), green bamboo (Sasa pygmaea), luxuriant and rich with fragrance white bamboo (Pleioblastus fortunei), floor file bamboos (Pleiblastus argenteastriatus).Group training research for drepanostachyum luodianense is less, culture media composition and application (201010268793.7) thereof that the sterilization method (201010268792.2) of current rarely seen explant in drepanostachyum luodianense tissue culture and drepanostachyum luodianense tissue are cultivated, these two contents are only the fast numerous process mid-early stage work of drepanostachyum luodianense, though drepanostachyum luodianense seedling growing process is had to important reference value, but cannot fundamentally solve the problem that karst drepanostachyum luodianense quantity sharply declines, in the sterilization method of explant in drepanostachyum luodianense tissue culture, (201010268792.2) method therefor is simultaneously: first soak with liquid detergent, rinse with flowing water afterwards, on superclean bench, use 70-75% alcohol disinfecting 1min, sterile water soaks rear with 0.1% mercuric chloride sterilizing 6-12 minute, the method is through checking, really effective to disinfecting of drepanostachyum luodianense explant, but because considering endurance and the disinfecting process impact on explant survival rate of explant on bactericide, although disinfecting time is longer, sterilization effect is better, but sterilization also may cause explant death for a long time, with drepanostachyum luodianense explant survival rate <30% after the method, thereby can not be simple judge best sterilization method according to pollution rate, should consider two aspects of pollution rate and survival rate simultaneously.In the culture media composition of cultivating at drepanostachyum luodianense tissue and application (201010268793.7) thereof, relate to the induction formula of drepanostachyum luodianense bud, but the induction of bud is only the beginning in the fast numerous drepanostachyum luodianense bamboo seedling of employing group training method, the propagation of later stage clump bud, take root, transplanting etc. is even more important.
Summary of the invention
Goal of the invention: for the deficiencies in the prior art, the object of this invention is to provide a kind of drepanostachyum luodianense method for quickly breeding, to overcome drepanostachyum luodianense breeding difficulty, alleviate the problem that under nature, drepanostachyum luodianense population quantity sharply reduces, fill up the blank of drepanostachyum luodianense in fast numerous.
Technical scheme: in order to realize foregoing invention object, the technical solution used in the present invention is as follows:
A kind of drepanostachyum luodianense method for quickly breeding, comprises the steps:
1) the drepanostachyum luodianense explant of getting, is placed in liquid detergent water and soaks 2min, and constantly shake, is then placed under flowing water and rinses 2-3h, blots surface moisture with filter paper, then, with 70% alcohol immersion 30 ~ 40s, with aseptic water washing for several times, immersion 0.1% HgCl
2in solution, process 3 ~ 8min, with aseptic water washing several, aseptic filter paper blots surface moisture;
2) drepanostachyum luodianense inducing clumping bud: the drepanostachyum luodianense explant after sterilization is transferred in medium mixture, 25 ± 1 DEG C, illumination 3000 lx, 14 h/d, induction drepanostachyum luodianense Multiple Buds, induction time continues 20-25d; Wherein, medium mixture: MS minimal medium, 6-BA concentration is 3 ~ 5mg/L, additional saccharose 30 g/L, as carbon source, add 6.5 g/L agar as coagulating agent; Regulating pH value with 1mol/LNaOH and 1mol/LHCl has been 5.8;
3) drepanostachyum luodianense adventitious buds proliferation: the Multiple Buds inducing is cut into the fritter with 3-5 bud on superclean bench, be transferred on proliferated culture medium mixture 25 ± 1 DEG C after separating, illumination 3000 lx, 14h/d, breeds cultivation, and propagation incubation time continues 15-30d; Wherein, proliferated culture medium mixture: MS minimal medium, growth regulator 6-BA concentration 3 ~ 5mg/L, KT concentration are 0.5 ~ 1mg/L, sucrose 30 g/L, and agar 6.5 g/L mix;
4) drepanostachyum luodianense Multiple Buds culture of rootage: by the drepanostachyum luodianense Multiple Buds after propagation, after separating, be transferred on culture of rootage based mixtures, 25 ± 1 DEG C, illumination 3000 lx, 14h/d, carries out culture of rootage, culture of rootage time remaining 50d-60d; Wherein, culture of rootage based mixtures: MS minimal medium, sucrose 30 g/L, agar 6.5 g/L, 6-BA concentration are that 1mg/L, NAA concentration are that 1 ~ 2mg/L, IBA concentration are 0.5 ~ 1mg/L.
5) drepanostachyum luodianense group training transplantation of seedlings: drepanostachyum luodianense group training seedling after culture of rootage 50-60d is carried out to hardening 7d, with running water, medium residual root system is rinsed well, plant in filling the nutritive cube of matrix.
In step 1), select robust growth and without the plant of damage by disease and insect, get wherein and be wrapped in intravaginal branch as drepanostachyum luodianense explant with semi-lignified, the stalk bud of do not sprout stalk bud or the stalk bud of just having sprouted.
In step 1), with 70% alcohol immersion 30s, use aseptic water washing 3 times, immerse 0.1% HgCl
2in solution, process 5min, use aseptic water washing 5 times.
Step 2) in, in medium mixture, 6-BA concentration is 5mg/L.
In step 3), in proliferated culture medium mixture, 6-BA concentration is that 5mg/L, KT concentration are 0.5mg/L.
In step 4), in culture of rootage based mixtures, NAA concentration is that 2mg/L, IBA concentration are 1mg/L.
In step 4), in culture of rootage based mixtures, NAA concentration is that 1mg/L, IBA concentration are 0.5mg/L.
In step 5), described matrix is vermiculite, and particle size is 2-4mm.
In step 5), described matrix is that volume ratio is the peat composed of rotten mosses of 1:1 and the mixture of vermiculite.
Beneficial effect: compared with prior art, drepanostachyum luodianense method for quickly breeding of the present invention, there is small investment, the drepanostachyum luodianense speed of growing seedlings is fast, survival rate is high, the advantages such as bamboo seedling growing way is consistent, all kinds of shortcomings that traditional parent bamboo produces are overcome, as low in reproductive efficiency, habitat destruction is large, breeding cycle is long, cost of transportation is high, and as a kind of drepanostachyum luodianense method for quickly breeding, adopt the method breeding drepanostachyum luodianense bamboo seedling not to be subject to seasonal restrictions, cost of transportation is cheap, being applicable to factorial seedling growth produces, for the quantity under fast quick-recovery drepanostachyum luodianense nature, performance drepanostachyum luodianense provides one approach fast and effectively in karst ecological benefits and economic benefit.
Brief description of the drawings
Fig. 1 is the stalk bud figure as explant;
Fig. 2 is the stalk bud figure of sprouting;
Fig. 3 is inducing clumping bud figure;
Fig. 4 is adventitious buds proliferation figure;
Fig. 5 is the Multiple Buds figure for taking root;
Fig. 6 is the regeneration plant figure of taking root;
Fig. 7 is new root situation map after transplanting 14d;
Fig. 8 is new root situation map after transplanting 24d;
Fig. 9 is the new root comparison diagram of different formulations after transplanting 44d.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.
The external sterilization of embodiment 1 drepanostachyum luodianense
The branch of semi-lignified (is explant: robust growth and without the plant of damage by disease and insect, get wherein and be wrapped in intravaginal branch with semi-lignified, the stalk bud of do not sprout stalk bud or the stalk bud of just having sprouted) by outdoor adopt back indoor after, be cut into segment, every section with a stalk bud, on joint, under joint, respectively stay 0.5-1cm, get white cat board liquid detergent 2-3ml, be dissolved in 2-3L running water, explant is positioned over and is placed on immersion 2min in liquid detergent water (concentration is about 1 ‰), and constantly shake, then be placed under flowing water and rinse 2-3h, blot surface moisture with filter paper.After pretreatment, stem section is positioned on superclean bench, with 70% ethanol and 0.1%HgCl
2solution carries out sterilization processing test to explant, after each ethanol disinfection processing, uses aseptic water washing 3 times, and mercuric chloride is used aseptic water washing 5 times after processing.
6 processing are established in this experiment altogether, and each processing repeats 5 times.Disinfection way 70% ethanol disinfection is processed 30s, 40s, 0.1% HgCl
2disinfect 3min, 5min, 8min.Stem section after sterilization is placed in sterilized culture dish, is inoculated into (initial medium is MS+6-BA3.0 mg/L) on initial medium, and 24 explants of each processing inoculation, repeat 3 times, add up pollution rate after 15d, after 25 d, add up survival rate.
Experimental result is as shown in table 1, from pollution rate, although the result of processing 6 is minimum, pollution rate is 1.4%, but its survival rate is also minimum, only have 41.7%, this processes drepanostachyum luodianense with ethanol 40 s and mercuric chloride 8 min with regard to explanation, although sterilization effect is fabulous, but also explant is caused to injury simultaneously, obviously reduce the survival rate of explant, as can be seen here, the processing method adopting in the sterilization method (201010268792.2) of document explant in drepanostachyum luodianense tissue culture, though pollution rate is low, but the survival rate of explant is also subject to very big impact, thereby processing 6 is not best sterilization method.
The different disinfectants of table 1 are processed the impact that different time is cultivated explant
Though using 70% ethanol disinfection 40s to process after stain rate than sterilization 30s decreases, but survival rate also significantly reduces, all lower than 50%, this is because drepanostachyum luodianense explant is tiny, children is tender, and ethanol has stronger penetration power and sterilizing power, process 40s drepanostachyum luodianense explant is existed to certain injury, thereby select 30s on the ethanol disinfection time.In the time that mercuric chloride disinfecting time is 3min, process 1 and all exceed 20% with the pollution rate of processing 4, be significantly higher than other processing, sterilization time is too short causes part germ not to be killed, thereby pollution rate is higher, can reach 61.1% though wherein process 1 survival rate, because its high pollution rate is not as optimization process.Processing 2 and process in 3 selection, although process 3 pollution rates a little less than processing 2, its survival rate is but significantly lower than processing 2, when its reason is sterilizing, and HgCl
2small part Hg dissociates
+, stalk bud is subject to Hg
+murder by poisoning and death has affected survival rate.Visible in the time reducing pollution rate measure, be not disinfecting time the longer the better, should not adopt the way of lengthening disinfection time.
Therefore,, according to above-mentioned result of the test, the best sterilization method of drepanostachyum luodianense explant is: flowing water rinses 2-3h → 70% Ethanol Treatment 30s → aseptic water washing 3 times → 0.1% HgCl
2process 5 times → aseptic filter paper of 5min → aseptic water washing and blot surface moisture.After the drepanostachyum luodianense explant inoculation after treatment of above sterilization method, aseptic rate is the highest, and bud induction survival rate is high.
Embodiment 2 drepanostachyum luodianense clump bud inductions
It is minimal medium that MS is selected in this test, use the generation of growth regulator 6-BA, NAA and KT combination induced bundle bud, the explant that the best sterilization method (embodiment 1) of selecting taking testing sieve was processed is induced as test material carries out clump bud, selects the hormone combination of the most applicable clump bud induction.24 bottles of every kind of concentration combination, 3 explants of every bottle graft kind.25 ± 1 DEG C, illumination 3000 lx, 14h/d, induces cultivation; Every 5d observes 1 time, and the observation cycle is 25d, adds up the induction number of clump bud, observes and record the growing state of bud.
The stalk bud of explant, as shown in Figure 1; Stalk bud is sprouted, as shown in Figure 2; Inducing clumping bud, as shown in Figure 3.Experimental result is as shown in table 2, and the optimization formula of drepanostachyum luodianense clump bud induction hormone combination combination is A
3b
1c
1, that is: MS+6-BA5 mg/L, without adding KT and NAA.Experimental data is compared known, the extreme difference of 6-BA is 29.48, the extreme difference of KT is 23.27, the extreme difference of NAA is 14.69, each hormone types on the impact of drepanostachyum luodianense clump bud induction rate is: 6-BA > KT > NAA, this explanation basic element of cell division is that clump bud is sprouted necessary.
The experimental result of table 2 clump bud induction hormone combination
Embodiment 3 drepanostachyum luodianense clump bud propagation
The clump bud inducing is cut into the fritter with several buds (3-5), after separating, be transferred on proliferated culture medium, 25 ± 1 DEG C, illumination 3000 lx, 14h/d, breeds cultivation.Use growth regulator 6-BA, KT, NAA combination to breed, experiment adopts Three factors-levels orthogonal design, and research variable concentrations combines the impact on successive propagation.
Adventitious buds proliferation, as shown in Figure 4.Experimental result is as shown in table 3, and the optimization formula of adventitious buds proliferation hormone combination combination is A
3b
2c
1, that is: MS+6-BA 5mg/L+KT 0.5mg/L.Experimental data is compared known, the extreme difference that the extreme difference that the extreme difference of 6-BA is 1.04, KT is 0.26, NAA is 0.47, between each hormone types, on the impact of adventitious buds proliferation multiple is: 6-BA > NAA > KT.
The experimental result of table 3 variable concentrations hormone to adventitious buds proliferation multiple
Embodiment 4 drepanostachyum luodianense clump buds are taken root
Train seedling as shown in Figure 5 for the drepanostachyum luodianense group of taking root, the drepanostachyum luodianense regrowth (cultivating through embodiment 2,3) of inducing, cultivating on shoot proliferation medium by the mode with the numerous bud of bud by stalk bud.Select healthy and strong drepanostachyum luodianense group training seedling, access root media, 25 ± 1 DEG C, illumination 3000 lx, 14h/d, inducing adventitious root.Taking MS as minimal medium, select growth hormone NAA, IBA and the combination of basic element of cell division 6-BA variable concentrations, carry out drepanostachyum luodianense group training seedling rooting culture studies, experimental design and each hormonal readiness are in table 4.
The orthogonal design scheme of table 4 regeneration plant culture of rootage
In process of the test, every 7d observes once, 5 bottles of drepanostachyum luodianense group training seedlings of random choose in each processing, group training seedling is taken out from blake bottle, carefully medium is cleaned up with running water, with scissors, root system is cut, added up root quantity, analyze each Root Characteristics value of each processing drepanostachyum luodianense group training seedling simultaneously with root system analyzer.
Experimental result is as shown in table 5, 9 kinds of formulas are to the processing of taking root of drepanostachyum luodianense group training seedling, wherein fill a prescription 2, formula 3 is processed at root length, root system volume, in the Root Characteristics indexs such as surface area, be obviously better than other formulas, adopt principal component analysis (PCA) to analyze 9 kinds of formulas, filter out a principal component, by principal component integrate score, find that formula 3 is for the optimization formula in the each root system formula of drepanostachyum luodianense group training seedling, but formula 2, 3 principal component scores values are comparatively approaching, therefore, drepanostachyum luodianense root system induces selectable formula to be: MS+6-BA 1mg/L+NAA 1mg/L+IBA 0.5mg/L, MS+6-BA 1mg/L+NAA 2mg/L+IBA 1mg/L.
The table 5 difference lower drepanostachyum luodianense group training of the formula shoot root of taking root is eigen value measurement result (mean value)
6-BA concentration is conducive to the growth with drepanostachyum luodianense root system most in the time of 1mg/L, and in the time that 6-BA concentration reaches 5mg/L, obviously drepanostachyum luodianense root growth is played to inhibitory action, made in 30d that the drepanostachyum luodianense group training seedling in P7, P8, P9 formula carries out in test without root growth.The regeneration plant of taking root as shown in Figure 6.
Embodiment 5 drepanostachyum luodianense group training transplantation of seedlings
Drepanostachyum luodianense group training transplantation of seedlings is key one ring of the fast numerous Establishing of drepanostachyum luodianense, after group training transplantation of seedlings, the height of survival rate and growing way quality are directly connected to the success or failure of the fast traditional font of drepanostachyum luodianense system, transplanted seedling should be well developed root system, robust growth, trains seedling without the group of damage by disease and insect, and the formulation selection in root system Induction Process is the prerequisite of growing way after decision drepanostachyum luodianense group training transplantation of seedlings as can be seen here.Known by analyzing drepanostachyum luodianense group training shoot root system induction experimental result, the formula of taking root is conducive to the root system induction of drepanostachyum luodianense for No. 2, No. 3 most, and root growth is the fastest, is ideal formula in drepanostachyum luodianense root system Induction Process.For further screening optimization formula and the best medium matter in the fast numerous Establishing of drepanostachyum luodianense, the present embodiment is chosen after the drepanostachyum luodianense group training seedling rooting of being cultivated for No. 2, No. 3 by the formula of taking root, and transplants.
Concrete grammar is as follows:
The drepanostachyum luodianense group training seedling rooting of growing in 2,3 medium at formula is processed to 40d, take off bottle cap, be positioned in plastic sack taking 12 bottles as one group, and in plastic sack, place one bottle of distilled water, fasten sack, to ensure the relative moisture of bag interior air.After 7 days, from blake bottle, carefully take out drepanostachyum luodianense group training seedling with tweezers, rinse the residual medium of root well with running water, plant in the nutritive cube that contains the peat composed of rotten mosses, vermiculite (2-4mm), the peat composed of rotten mosses+vermiculite (1:1 by volume) culture matrix, the specification of nutritive cube is 20cm × 20cm, 20 basins are established in each processing, are placed in green house, and the moisturizing of often spraying water.Transplant after 20d, 5 basins are got in every processing, and transplanted seedling is taken out, and be placed in clear water the matrix being attached on root system is cleaned, and with the newborn root system of scissors clip (new root color is milky), by root system analysis-e/or determining root associated eigenvalue, analyze the upgrowth situation of root system.
Experimental result is as follows:
Two kinds of formulas are all the longest with the drepanostachyum luodianense group training seedling root length of vermiculite cultivation, take second place, with drepanostachyum luodianense group training seedling root length minimum in peat composed of rotten mosses formula with the peat composed of rotten mosses+vermiculite formula.Vermiculite is cultivated No. 2 formula drepanostachyum luodianense group training seedling root lengths and is improved 65.72% compared with the root system of peat composed of rotten mosses cultivation, and vermiculite is cultivated No. 3 formula drepanostachyum luodianense group training seedling root lengths compared with the root system 31.01% of peat composed of rotten mosses cultivation.Two kinds of formulas are all that growth rate is the fastest with the drepanostachyum luodianense group training shoot root of vermiculite cultivation, are 28.65cm/d, and cultivating drepanostachyum luodianense group training shoot root with the peat composed of rotten mosses is that growth rate is minimum, is 19.29 cm/d.The drepanostachyum luodianense group training shoot root of cultivating with vermiculite in 3 kinds of matrix formulations is surface area maximum, is surface area minimum with drepanostachyum luodianense group training shoot root in peat composed of rotten mosses formula, and vermiculite formula improves 45.24% compared with peat composed of rotten mosses formula.In No. 2 formulas, in vermiculite matrix and the peat composed of rotten mosses+vermiculite matrix, root system surface area is all higher than No. 3 formulas, in vermiculite formula, No. 2 formulas are high by 18.08% compared with No. 3, No. 2 the formula peat composed of rotten mosses+vermiculite matrix improves 11.72% compared with No. 3, and in peat composed of rotten mosses matrix, No. 3 formula drepanostachyum luodianense group training shoot roots are that surface area will, higher than No. 2 formulas, improve 12.48%.Drepanostachyum luodianense group training seedling root system volume is the fastest with No. 2 peat composed of rotten mosses+vermiculite rates of climb, and its rate of rise is 0.066 cm
3/ d, the slowest with No. 2 peat composed of rotten mosses root system volume rates of rise, be only 44.21% of No. 2 peat composed of rotten mosses+vermiculites.In 3 kinds of matrix formulations, with drepanostachyum luodianense root system volume maximum in the peat composed of rotten mosses+vermiculite formula, minimum with peat composed of rotten mosses formula, be only 67.64% of the peat composed of rotten mosses+vermiculite formula.Take root in formula for 2 kinds, in No. 2 vermiculites, the peat composed of rotten mosses+vermiculite matrix, root system volume is all higher than No. 3, and in peat composed of rotten mosses matrix, taking root for No. 3 formula will, higher than No. 2, improve 30.87%.Respectively take root, in matrix formulations, with No. 2 formula root system fractal dimension maximums in vermiculite of taking root, be 1.56, minimum in the peat composed of rotten mosses with No. 2 formulas of taking root, be only 92.93% of vermiculite.Take root in formula, in peat composed of rotten mosses matrix, No. 3 formula will be higher than No. 2, and at vermiculite, the peat composed of rotten mosses+vermiculite matrix, the formula of taking root for No. 3, all lower than No. 2, is respectively 97.46%, 98.78% of No. 2 formulas.
As can be seen here, vermiculite treatment effect is better than the peat composed of rotten mosses, the peat composed of rotten mosses+vermiculite, and survival rate reaches 100%, thereby the optimal selection of transplanting medium is vermiculite.Cause the reason of growth differences of this class matrix container seedling relevant with the hormone prescription adopting in early stage group training seedling rooting process simultaneously, result shows, after the drepanostachyum luodianense group training transplantation of seedlings of 2 processing of filling a prescription, root growth situation is better than No. 3, the interpretation of result of comprehensive drepanostachyum luodianense group training seedling rooting treatment formulations, P2 formula is trained seedling rooting to comparatively desirable formula, i.e. MS+6-BA 1 mg/L+NAA 1mg/L+IBA 0.5 mg/L in transplanting process for drepanostachyum luodianense group.
New root situation after transplanting 14d, as shown in Figure 7; New root situation after transplanting 24d, as shown in Figure 8; After transplanting 44d, the new root of different formulations contrasts, as shown in Figure 9.
The above, it is only preferred embodiment of the present invention, in the present invention, the content such as related liquid detergent brand is not done any pro forma restriction to the present invention, any technical solution of the present invention content that do not depart from, any simple modification, equivalent variations and the modification above embodiment done according to technical spirit of the present invention, all belong to the scope of technical solution of the present invention.
Claims (9)
1. a drepanostachyum luodianense method for quickly breeding, is characterized in that, comprises the steps:
1) the drepanostachyum luodianense explant of getting, is placed in liquid detergent water and soaks 2min, and constantly shake, is then placed under flowing water and rinses 2-3h, blots surface moisture with filter paper, then, with 70% alcohol immersion 30 ~ 40s, with aseptic water washing for several times, immersion 0.1% HgCl
2in solution, process 3 ~ 8min, with aseptic water washing several, aseptic filter paper blots surface moisture;
2) drepanostachyum luodianense inducing clumping bud: the drepanostachyum luodianense explant after sterilization is transferred in medium mixture, 25 ± 1 DEG C, illumination 3000 lx, 14 h/d, induction drepanostachyum luodianense Multiple Buds, induction time continues 20-25d; Wherein, medium mixture: MS minimal medium, 6-BA concentration is 3 ~ 5mg/L, additional saccharose 30 g/L, as carbon source, add 6.5 g/L agar as coagulating agent; Regulating pH value with 1mol/LNaOH and 1mol/LHCl is 5.8;
3) drepanostachyum luodianense adventitious buds proliferation: the Multiple Buds inducing is cut into the fritter with 3-5 bud on superclean bench, be transferred on proliferated culture medium mixture 25 ± 1 DEG C after separating, illumination 3000 lx, 14h/d, breeds cultivation, and propagation incubation time continues 15-30d; Wherein, proliferated culture medium mixture: MS minimal medium, growth regulator 6-BA concentration 3 ~ 5mg/L, KT concentration are 0.5 ~ 1mg/L, sucrose 30 g/L, and agar 6.5 g/L mix;
4) drepanostachyum luodianense Multiple Buds culture of rootage: by the drepanostachyum luodianense Multiple Buds after propagation, after separating, be transferred on culture of rootage based mixtures, 25 ± 1 DEG C, illumination 3000 lx, 14h/d, carries out culture of rootage, culture of rootage time remaining 50d-60d; Wherein, culture of rootage based mixtures: MS minimal medium, sucrose 30 g/L, agar 6.5 g/L, 6-BA concentration are that 1mg/L, NAA concentration are that 1 ~ 2mg/L, IBA concentration are 0.5 ~ 1mg/L;
5) drepanostachyum luodianense group training transplantation of seedlings: drepanostachyum luodianense group training seedling after culture of rootage 50-60d is carried out to hardening 7d, with running water, medium residual root system is rinsed well, plant in filling the nutritive cube of matrix.
2. drepanostachyum luodianense method for quickly breeding according to claim 1, it is characterized in that, in step 1), select robust growth and without the plant of damage by disease and insect, get wherein and be wrapped in intravaginal branch as drepanostachyum luodianense explant with semi-lignified, the stalk bud of do not sprout stalk bud or the stalk bud of just having sprouted.
3. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that, in step 1), with 70% alcohol immersion 30s, uses aseptic water washing 3 times, immerses 0.1% HgCl
2in solution, process 5min, use aseptic water washing 5 times.
4. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that step 2) in, in medium mixture, 6-BA concentration is 5mg/L.
5. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that, in step 3), in proliferated culture medium mixture, 6-BA concentration is that 5mg/L, KT concentration are 0.5mg/L.
6. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that, in step 4), in culture of rootage based mixtures, NAA concentration is that 2mg/L, IBA concentration are 1mg/L.
7. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that, in step 4), in culture of rootage based mixtures, NAA concentration is that 1mg/L, IBA concentration are 0.5mg/L.
8. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that, in step 5), described matrix is vermiculite.
9. drepanostachyum luodianense method for quickly breeding according to claim 1, is characterized in that, in step 5), described matrix is that volume ratio is the peat composed of rotten mosses of 1:1 and the mixture of vermiculite.
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| CN104705186A (en) * | 2015-03-10 | 2015-06-17 | 朱海燕 | Tissue culture and quick propagation method of S. versicolor |
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Application publication date: 20130925 Assignee: Nanyue agricultural and Forestry Development Research Institute of Yixing City Assignor: Nanjing Forestry University Contract record no.: X2019320000137 Denomination of invention: Rapidly breeding method for ampelocalamus luodianensis Granted publication date: 20141210 License type: Common License Record date: 20191021 Application publication date: 20130925 Assignee: Yangzhou Dayu scenic bamboo garden Assignor: Nanjing Forestry University Contract record no.: X2019320000138 Denomination of invention: Rapidly breeding method for ampelocalamus luodianensis Granted publication date: 20141210 License type: Common License Record date: 20191021 |