CN103314848A - 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) be the outstanding Sinobambusa of grass family Bambusoideae (
Ampelocalamus) the bamboo kind, be the endangered plants of karst, mainly be 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 ', the vertical distribution height is between 650-1250m.Its good looking appearance, the cultivation ornamental value is arranged, simultaneously, drepanostachyum luodianense can be at the abominable 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 obviously is better than fruticeta to the interception of rainfall, the quantity of water-stable aggregate also is higher than fruticeta and meadow in the sylvan life soil, in Karst ecosystem, the ecological functions such as water source self-restraint, water and soil conservation, nutrient balance are played an important role, have 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 drepanostachyum luodianense is listed in utmost point danger species in " Chinese species Red List (first volume: Red List) ".
The bamboo plant blooming cycle is long, be difficult to take the mode of sexual propagation to obtain the bamboo seedling, at present the modes of reproduction of bamboo plant mainly adopts and moves bamboo, buries whip, the modes of reproduction such as cuttage obtains the 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 the 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 the cuttage and seedling culture reproduction coefficient before dual mode high, be subject to seasonal restrictions in the branch cutting process 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 efficient is extremely low, can't effectively alleviate the problem that the present population quantity of drepanostachyum luodianense sharply reduces.The method efficient 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, be fit to the batch production production of drepanostachyum luodianense bamboo seedling, can fundamentally alleviate the problem that the drepanostachyum luodianense population quantity descends.
Up to now, Bamboo Tissue training both at home and abroad makes great progress, by tissue culture technique, realized to the safety bamboo (
Pseudosasa japonica ' Tsutsumiana'), black bamboo (
Phyllostachys nigra), the gold inlaid jade bamboo (
Phyllostachys aureosulcata ' Spectabilis'), green bamboo (
Sasa pygmaea), luxuriant and rich with fragrance white bamboo (
Pleioblastus fortunei), the floor file bamboo (
Pleiblastus argenteastriatus) etc. the tissue-culturing rapid propagation of Ornamental Bamboo.Group training research for drepanostachyum luodianense is less, culture media composition and application (201010268793.7) thereof that the sterilization method of present rarely seen explant in drepanostachyum luodianense tissue culture (201010268792.2) and drepanostachyum luodianense tissue are cultivated, these two contents only are the mid-early stage work of the fast numerous process of drepanostachyum luodianense, though the drepanostachyum luodianense seedling growing process is had important reference value, but can't fundamentally solve the problem that karst drepanostachyum luodianense quantity sharply descends, (201010268792.2) method therefor is in the sterilization method of explant in drepanostachyum luodianense tissue culture simultaneously: soak with liquid detergent first, wash with flowing water afterwards, at superclean bench 70-75% alcohol disinfecting 1min, after soaking, sterile water sterilized 6-12 minute with 0.1% mercuric chloride, the method is through checking, really effective to disinfecting of drepanostachyum luodianense explant, but because considering that explant is on the impact on the explant survival rate of the endurance of bactericide and disinfecting process, although disinfecting time is longer, sterilization effect is better, but sterilization also may cause explant dead 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 simultaneously two aspects of pollution rate and survival rate.In culture media composition that the drepanostachyum luodianense tissue is cultivated and application (201010268793.7) thereof, related to drepanostachyum luodianense spore induction prescription, but spore induction only is 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 purpose 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 the drepanostachyum luodianense population quantity sharply reduces under the nature, fill up the blank of drepanostachyum luodianense in fast numerous.
Technical scheme: in order to realize the foregoing invention purpose, 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 the liquid detergent water and soaks 2min, and constantly shakes, and then is placed under the flowing water and washes 2-3h, blots surface moisture with filter paper, then, with 70% alcohol immersion, 30 ~ 40s, with aseptic water washing for several times, immerses 0.1% HgCl
2Process 3 ~ 8min in the solution, with aseptic water washing for several times, aseptic filter paper blots surface moisture;
2) drepanostachyum luodianense inducing clumping bud: the drepanostachyum luodianense explant after will sterilizing is transferred in the medium mixture, and 25 ± 1 ℃, illumination 3000 lx, 14 h/d induce the drepanostachyum luodianense Multiple Buds, and induction time continues 20-25d; Wherein, the medium mixture: MS minimal medium, 6-BA concentration are 3 ~ 5mg/L, and additional saccharose 30 g/L add 6.5 g/L agar as coagulating agent as carbon source; Regulating the pH value with 1mol/LNaOH and 1mol/LHCl has been 5.8;
3) drepanostachyum luodianense adventitious buds proliferation: the Multiple Buds that induces is cut into fritter with 3-5 bud at superclean bench, after separating, be transferred on the proliferated culture medium mixture 25 ± 1 ℃, illumination 3000 lx, 14h/d breeds cultivation, and the 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: the drepanostachyum luodianense Multiple Buds after will breeding, after separating, be transferred on the culture of rootage based mixtures, 25 ± 1 ℃, 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 behind the culture of rootage 50-60d is carried out hardening 7d, rinse well with the medium that running water is residual with root system, plant in the nutritive cube that fills matrix.
In the step 1), select robust growth and without the plant of damage by disease and insect, semi-lignified, the stalk bud got wherein with do not sprout stalk bud or the stalk bud of just having sprouted are wrapped in intravaginal branch as the drepanostachyum luodianense explant.
In the step 1), with 70% alcohol immersion 30s, use aseptic water washing 3 times, immerse 0.1% HgCl
2Process 5min in the solution, use aseptic water washing 5 times.
Step 2) in, in the medium mixture, 6-BA concentration is 5mg/L.
In the step 3), in the proliferated culture medium mixture, 6-BA concentration is that 5mg/L, KT concentration are 0.5mg/L.
In the step 4), in the culture of rootage based mixtures, NAA concentration is that 2mg/L, IBA concentration are 1mg/L.
In the step 4), in the culture of rootage based mixtures, NAA concentration is that 1mg/L, IBA concentration are 0.5mg/L.
In the step 5), described matrix is vermiculite, and particle size is 2-4mm.
In the 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, has 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 have been overcome, low such as 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, be fit to factorial seedling growth production, be the quantity under the fast quick-recovery drepanostachyum luodianense nature, the performance drepanostachyum luodianense provides a kind of fast and effectively approach in karst ecological benefits and economic benefit.
Description of 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 be used to the Multiple Buds figure of taking root;
Fig. 6 is the regeneration plant figure of taking root;
Fig. 7 is new root situation map behind the transplanting 14d;
Fig. 8 is new root situation map behind the transplanting 24d;
Fig. 9 is the new root comparison diagram of different formulations behind the 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, semi-lignified, the stalk bud got wherein with do not sprout stalk bud or the stalk bud of just having sprouted are wrapped in intravaginal branch) by outdoor adopt back indoor after, be cut into segment, every section with a stalk bud, the lower 0.5-1cm that respectively stays of joint on the joint, get white cat board liquid detergent 2-3ml, be dissolved in the 2-3L running water, explant is positioned over to be placed on soaks 2min in the liquid detergent water (concentration is about 1 ‰), and constantly shake, then be placed under the flowing water and wash 2-3h, blot surface moisture with filter paper.After the preliminary treatment, the stem section is positioned on the superclean bench, with 70% ethanol and 0.1%HgCl
2Solution carries out the sterilization Processing Test to explant, uses aseptic water washing 3 times after each ethanol disinfection processing, uses aseptic water washing 5 times after mercuric chloride is processed.
6 processing are established in this experiment altogether, and each is processed and repeats 5 times.Disinfection way 70% ethanol disinfection is processed 30s, 40s, 0.1% HgCl
2Disinfect 3min, 5min, 8min.Stem section after the sterilization is placed in the sterilized culture dish, is inoculated into (initial medium is MS+6-BA3.0 mg/L) on the initial medium, and each processes 24 explants of inoculation, repeats 3 times, adds up pollution rate behind the 15d, adds up survival rate behind 25 d.
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, also explant has been caused injury simultaneously, obviously reduced the survival rate of explant, this shows, the processing method that adopts in the sterilization method of document explant in drepanostachyum luodianense tissue culture (201010268792.2) is though pollution rate is low, but the survival rate of explant also is subject to very big impact, thereby to process 6 be 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 the after stain rate than sterilization 30s decreases, but survival rate also significantly reduces, all be lower than 50%, this is because the drepanostachyum luodianense explant is tiny, the children is tender, and ethanol has stronger penetration power and sterilizing power, process 40s there is certain injury in the drepanostachyum luodianense explant, thereby select 30s in the ethanol disinfection time.When the mercuric chloride disinfecting time is 3min, the pollution rate of processing 1 and processing 4 is all above 20%, be significantly higher than other processing, sterilization time is too short to cause the part germ not to be killed, thereby pollution rate is higher, can reach 61.1% though wherein process 1 survival rate, because of its high pollution rate not as optimization process.Processing 2 and process in 3 the selection, although process 3 pollution rates a little less than processing 2, its survival rate but significantly is lower than processes 2, when its reason is sterilization, and HgCl
2Small part Hg dissociates
+, the stalk bud is subject to Hg
+Murder by poisoning and death has affected survival rate.As seen when reducing the 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 flushing 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 the above sterilization method processing, aseptic rate is the highest, and bud induces survival rate high.
Embodiment 2 drepanostachyum luodianense clump buds are induced
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 take testing sieve was processed is induced as test material carries out clump bud, the hormone combination of selecting the most suitable clump bud to induce.24 bottles of every kind of concentration combination, 3 explants of every bottle graft kind.25 ± 1 ℃, illumination 3000 lx, 14h/d induces cultivation; Every 5d observes 1 time, and the observation cycle is 25d, statistics clump spore induction number, the growing state of observation and record bud.
The stalk bud of explant, as shown in Figure 1; The 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 it is A that drepanostachyum luodianense clump bud is induced the optimization formula of hormone combination combination
3B
1C
1, that is: MS+6-BA5 mg/L need not to add KT and NAA.Experimental data is compared as can be known, the extreme difference of 6-BA is that the extreme difference of 29.48, KT is that the extreme difference of 23.27, 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.
Table 2 clump bud is induced the experimental result of hormone combination
Embodiment 3 drepanostachyum luodianense clump buds propagation
The clump bud that induces is cut into fritter with several buds (3-5), after separating, be transferred on the proliferated culture medium, 25 ± 1 ℃, illumination 3000 lx, 14h/d breeds cultivation.Use growth regulator 6-BA, KT, NAA combination to breed, the Three factors-levels orthogonal design is adopted in experiment, and the combination of research variable concentrations is on the impact of 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 as can be known, and the extreme difference of 6-BA is that the extreme difference of 1.04, KT is that the extreme difference of 0.26, NAA is 0.47, between each hormone types on the impact of adventitious buds proliferation multiple is: the 6-BA＞NAA＞KT.
Table 3 variable concentrations hormone is to the experimental result of adventitious buds proliferation multiple
Embodiment 4 drepanostachyum luodianense clump buds are taken root
Be used for the drepanostachyum luodianense group training seedling take root as shown in Figure 5, by the stalk bud induce, drepanostachyum luodianense regrowth (cultivating through embodiment 2,3) by cultivating at the shoot proliferation medium in the mode of the numerous bud of bud.Select healthy and strong drepanostachyum luodianense group training seedling, access root media, 25 ± 1 ℃, illumination 3000 lx, 14h/d, inducing adventitious root.Take MS as minimal medium, select the combination of growth hormone NAA, IBA and 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 see Table 4.
The orthogonal design scheme of table 4 regeneration plant culture of rootage
In the process of the test, every 7d observes once, 5 bottles of drepanostachyum luodianense group trainings of random choose seedling during each is processed, to organize the training seedling takes out from blake bottle, carefully medium is cleaned up with running water, with scissors root system is cut, the statistics root quantity is respectively processed each Root Characteristics value that the drepanostachyum luodianense group is trained seedling with the analysis of root system analyzer simultaneously.
Experimental result is as shown in table 5,9 kinds of prescriptions are to the processing of taking root of drepanostachyum luodianense group training seedling, wherein fill a prescription 2, prescription 3 is processed at root length, root system volume, obviously be better than other prescriptions on the Root Characteristics indexs such as surface area, adopt principal component analysis (PCA) that 9 kinds of prescriptions are analyzed, filter out a principal component, by the principal component integrate score, find that prescription 3 is the optimization formula in each root system prescription of drepanostachyum luodianense group training seedling, but fill a prescription 2,3 principal component scores values are comparatively approaching, therefore, the drepanostachyum luodianense root system induces selectable prescription 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.
Drepanostachyum luodianense group training shoot root was eigen value measurement result (mean value) under table 5 difference was taken root and filled a prescription
6-BA concentration is conducive to and the drepanostachyum luodianense Root growth when 1mg/L most, and when 6-BA concentration reaches 5mg/L, obviously the drepanostachyum luodianense root growth has been played inhibitory action, the drepanostachyum luodianense group training seedling in P7, P8, the P9 prescription is grown without root system in the 30d that test is carried out.The regeneration plant of taking root as shown in Figure 6.
Embodiment 5 drepanostachyum luodianense groups training transplantation of seedlings
Drepanostachyum luodianense group training transplantation of seedlings is key one ring that the fast traditional font of drepanostachyum luodianense system sets up, 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 after the group training transplantation of seedlings, transplanted seedling should be well developed root system, robust growth, trains seedling without the group of damage by disease and insect, this shows that the formulation selection in the root system Induction Process is the prerequisite of growing way after the decision drepanostachyum luodianense group training transplantation of seedlings.Induce experimental result as can be known by analyzing drepanostachyum luodianense group training shoot root system, the prescription of taking root is conducive to the root system of drepanostachyum luodianense for No. 2, No. 3 most induces, and root growth is the fastest, is ideal prescription in the drepanostachyum luodianense root system Induction Process.Be optimization formula and best medium matter during further the fast traditional font of screening drepanostachyum luodianense system sets up, the present embodiment is transplanted after choosing the drepanostachyum luodianense group training seedling rooting of being cultivated for No. 2, No. 3 by the prescription of taking root.
Concrete grammar is as follows:
To process 40d at the drepanostachyum luodianense group training seedling rooting that prescription be grown in 2,3 medium, take off bottle cap, and be positioned in the plastic sack take 12 bottles as one group, and in plastic sack, place one bottle of distilled water, fasten sack, to guarantee the relative moisture of air in the bag.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 (by volume 1:1) culture matrix, the specification of nutritive cube is 20cm * 20cm, 20 basins are established in each processing, place in the green house, and the moisturizing of often spraying water.After transplanting 20d, 5 basins are got in every processing, and transplanted seedling is taken out, and the matrix that places clear water will be attached on the root system is cleaned, and with the newborn root system of scissors clip (new root color is milky), with root system analysis-e/or determining root associated eigenvalue, analyze the Root growth situation.
Experimental result is as follows:
Two kinds of prescriptions are that length is the longest with the drepanostachyum luodianense group training shoot root that vermiculite is cultivated all, take second place with the peat composed of rotten mosses+vermiculite prescription, and be that length is minimum with drepanostachyum luodianense group training shoot root in the peat composed of rotten mosses prescription.It is that the root system that length is cultivated than the peat composed of rotten mosses improves 65.72% that vermiculite is cultivated No. 2 prescription drepanostachyum luodianense group training shoot roots, and it is the root system 31.01% that length is cultivated than the peat composed of rotten mosses that vermiculite is cultivated No. 3 prescription drepanostachyum luodianense group training shoot roots.Two kinds of prescriptions are that growth rate is the fastest with the drepanostachyum luodianense group training shoot root that vermiculite is cultivated all, 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.Training shoot root with the drepanostachyum luodianense group of vermiculite cultivation in 3 kinds of matrix formulations is that surface area is maximum, is that surface area is minimum with drepanostachyum luodianense group training shoot root in the peat composed of rotten mosses prescription, and the vermiculite prescription is than peat composed of rotten mosses prescription raising 45.24%.In No. 2 prescriptions, the root system surface area all is higher than prescription No. 3 in vermiculite matrix and the peat composed of rotten mosses+vermiculite matrix, No. 2 prescriptions are high by 18.08% than No. 3 in the vermiculite prescription, No. 2 the prescription peat composed of rotten mosses+vermiculite matrix improves 11.72% than No. 3, and in the peat composed of rotten mosses matrix, No. 3 prescription drepanostachyum luodianense group training shoot root is that surface area will be higher than prescription No. 2, improves 12.48%.Drepanostachyum luodianense group training shoot root is that 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, only be 44.21% of No. 2 peat composed of rotten mosses+vermiculites.In 3 kinds of matrix formulations, maximum with drepanostachyum luodianense root system volume in the peat composed of rotten mosses+vermiculite prescription, minimum with peat composed of rotten mosses prescription, only be 67.64% of the peat composed of rotten mosses+vermiculite prescription.In 2 kinds of prescriptions of taking root, root system volume all is higher than No. 3 in No. 2 vermiculites, the peat composed of rotten mosses+vermiculite matrix, and in the peat composed of rotten mosses matrix, No. 3 the prescription of taking root will be higher than No. 2, improves 30.87%.Respectively take root, in the matrix formulations, maximum with No. 2 prescription root system fractal dimensions in vermiculite of taking root, be 1.56, minimum in the peat composed of rotten mosses with No. 2 prescriptions of taking root, only be 92.93% of vermiculite.Take root in the prescription, in the peat composed of rotten mosses matrix, No. 3 prescription will be higher than No. 2, and at vermiculite, the peat composed of rotten mosses+vermiculite matrix, No. 3 the prescription of taking root all is lower than No. 2, is respectively 97.46%, 98.78% of No. 2 prescriptions.
This shows, the 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 simultaneously the hormone prescription that adopts in reason and the early stage group training seedling rooting process of growth differences of this class matrix container seedling relevant, the result shows, the root growth situation is better than No. 3 after the 2 drepanostachyum luodianense group training transplantation of seedlings of processing of filling a prescription, the interpretation of result of comprehensive drepanostachyum luodianense group training seedling rooting treatment formulations, P2 prescription is trained seedling rooting comparatively desirable prescription, i.e. MS+6-BA 1 mg/L+NAA 1mg/L+IBA 0.5 mg/L in the transplanting process for the drepanostachyum luodianense group.
New root situation behind the transplanting 14d, as shown in Figure 7; New root situation behind the transplanting 24d, as shown in Figure 8; The new root of different formulations contrasts after transplanting 44d, as shown in Figure 9.
The above, it only is preferred embodiment of the present invention, the content such as related liquid detergent brand is not that the present invention is done any pro forma restriction among the present invention, any technical solution of the present invention content that do not break away from, to any simple modification, equivalent variations and modification that above embodiment does, all belong to the scope of technical solution of the present invention according to technical spirit 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 the liquid detergent water and soaks 2min, and constantly shakes, and then is placed under the flowing water and washes 2-3h, blots surface moisture with filter paper, then, with 70% alcohol immersion, 30 ~ 40s, with aseptic water washing for several times, immerses 0.1% HgCl
2Process 3 ~ 8min in the solution, with aseptic water washing for several times, aseptic filter paper blots surface moisture;
2) drepanostachyum luodianense inducing clumping bud: the drepanostachyum luodianense explant after will sterilizing is transferred in the medium mixture, and 25 ± 1 ℃, illumination 3000 lx, 14 h/d induce the drepanostachyum luodianense Multiple Buds, and induction time continues 20-25d; Wherein, the medium mixture: MS minimal medium, 6-BA concentration are 3 ~ 5mg/L, and additional saccharose 30 g/L add 6.5 g/L agar as coagulating agent as carbon source; Regulating the pH value with 1mol/LNaOH and 1mol/LHCl has been 5.8;
3) drepanostachyum luodianense adventitious buds proliferation: the Multiple Buds that induces is cut into fritter with 3-5 bud at superclean bench, after separating, be transferred on the proliferated culture medium mixture 25 ± 1 ℃, illumination 3000 lx, 14h/d breeds cultivation, and the 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: the drepanostachyum luodianense Multiple Buds after will breeding, after separating, be transferred on the culture of rootage based mixtures, 25 ± 1 ℃, illumination 3000 lx, 14h/d carries out culture of rootage, culture of rootage time remaining 50d-60d; Wherein, the 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 behind the culture of rootage 50-60d is carried out hardening 7d, rinse well with the medium that running water is residual with root system, plant in the nutritive cube that fills matrix.
2. drepanostachyum luodianense method for quickly breeding according to claim 1, it is characterized in that, in the step 1), select robust growth and without the plant of damage by disease and insect, semi-lignified, the stalk bud got wherein with do not sprout stalk bud or the stalk bud of just having sprouted are wrapped in intravaginal branch as the drepanostachyum luodianense explant.
3. drepanostachyum luodianense method for quickly breeding according to claim 1 is characterized in that, in the step 1), with 70% alcohol immersion 30s, uses aseptic water washing 3 times, immerses 0.1% HgCl
2Process 5min in the solution, 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 the medium mixture, 6-BA concentration is 5mg/L.
5. drepanostachyum luodianense method for quickly breeding according to claim 1 is characterized in that, in the step 3), in the 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 the step 4), in the 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 the step 4), in the 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 the step 5), described matrix is vermiculite.
9. drepanostachyum luodianense method for quickly breeding according to claim 1 is characterized in that, in the 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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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 |