CN105123519A - Ground cover grevillea tissue culturing method - Google Patents
Ground cover grevillea tissue culturing method Download PDFInfo
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- CN105123519A CN105123519A CN201510550899.9A CN201510550899A CN105123519A CN 105123519 A CN105123519 A CN 105123519A CN 201510550899 A CN201510550899 A CN 201510550899A CN 105123519 A CN105123519 A CN 105123519A
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Abstract
The invention discloses a ground cover grevillea tissue culturing method. The method includes the steps that a disinfected explant is inoculated to clumpy bud induction culturing media, and clumpy buds are induced; the clumpy buds are inoculated to subculturing enrichment culturing media to be continuously cultured; then the clump buds are inoculated to rooting culturing media to root, and seedling hardening and field culturing management are carried out after rooting is achieved. Ground cover grevillea clone seedlings obtained with the method are high in yield and short in time, and the obtained seedlings are robust and solve the problem that ground cover grevillea is lacked in the planting industry.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding, be specifically related to a kind ofly by the method for silvery birch tissue cultures.
Background technology
Silvery birch (Grevillea) is that Proteaceae (Proteaceae) can burst forth the plant of large-scale inflorescence, originates in the Sulawesi Island in the middle part of Australia, New Guinea, New Caledonia and Indonesia.Silvery birch is the third-largest seeds closely inferior to yearning between lovers and eucalyptus in Australian native plants, and have a very wide distribution, the desert area from the tropical rain forest of year rainfall 3000mm to year rainfall 200mm, has distribution from high mountain to coastal and arid inland.Genotype is varied, about 360 kinds, variety variations from the ground being less than 0.5m by silvery birch to 1 ~ 5m shrub and the arbor up to 35m.Silvery birch easily realizes interspecific cross and natural variation, therefore naturally produces or has artificially cultivated hundreds of cultivar and crossbreed.
Ground is famous drought-resistant seeds by silvery birch, is mostly distributed in Australia, western desert area, and the long-term drought in this area, because this place is had very strong drought-resistant ability by a lot of kinds of silvery birch.There are many tiny intensive fibrous roots on ground in silvery birch root structure, this is also long term survival in barren on the spot lower evolution result, and such structure is conducive to the nutrient in absorption lean soil, because this place is also barren-resistant seeds by silvery birch.Plantation ground is only needed by silvery birch the slow-release fertilizer executing low fertilizer efficiency, or does not apply fertilizer.Therefore, ground is had by silvery birch and manages and protects the low feature of cost.
Ground is raceme by silvery birch, and its flower pattern is peculiar, mostly in hairbrush shape or spider shape, flower shape is large, pattern is many, and have from blood orange yellow to yellow fraction, milky white even green, the florescence is long, some kinds are bloomed the whole year, ground is leaf abundant by silvery birch, and from needle-like, blade is shallow to be split and drastic crack, leaf look mostly is dark green, grayish green, and gardening has very high ornamental value.Ground is applicable to intensity by silvery birch and prunes, and plastic to produce the gardening come in every shape tree-like.Ground is high by survival rate in silvery birch grafting to the stock of shrub silvery birch and arbor silvery birch, can cultivate novel weeping branch silvery birch, because this place is widely used in courtyard greening and landscape engineering by Australia by silvery birch.Ground containing abundant nectar, can be provided food for the mankind and birds, honeybee class by silvery birch, because of the seeds that this place is famous trick bird by silvery birch and keeps pet.
China at present a large amount of grown place is also immature by the technology of silvery birch nursery stock, and be difficult to accomplish the intensive of operation, the high efficiency of production, also cannot meet market over the ground by the wilderness demand of silvery birch seedling.The ground that method for building up is simple, rooting rate is high, survival rate is high, by silvery birch method for tissue culture, will be played important effect by applying of silvery birch over the ground.
Summary of the invention
In order to solve above-mentioned Problems existing, the present invention has carried out ground by silvery birch Study on tissue culture under the support of introduce projects " introduced by silvery birch new varieties and raising technology ", establish ripe ground by the technological process of silvery birch tissue culture technique platform and factorial seedling growth, and successful reproduction 30,000 strains ground by silvery birch plantlet in vitro.
The object of the present invention is to provide a kind ofly by the method for silvery birch tissue cultures.
The technical solution used in the present invention is:
By a method for silvery birch tissue cultures, comprise the following steps:
1) collection of explant;
2) sterilization of explant;
3) clump bud inducement is cultivated: be seeded in clump bud inducement medium by the explant after upper step sterilization, Fiber differentiation 3 ~ 5 months, period need be forwarded to new clump bud inducement medium 3 ~ 5 times, and explant can be induced out multiple clumps of buds;
4) shoot proliferation is cultivated: the ground upper one-step inducing being gone out clump bud in subculture multiplication medium by silvery birch tissue culture plant inoculation, is often cultivated to be forwarded in new subculture multiplication medium by silvery birch propagation seedling on ground for 28 ~ 32 days and continued to cultivate;
5) root induction is cultivated: cut by the simple bud highly reaching more than 2cm in Multiplying culture in access root media, cultivate 10 ~ 15 days;
6) hardening: enter the hardening stage after the propagation seedling when at least 60% starts to take root, is taken root the ground of acquisition for plantlet in vitro light intensity 3000 ~ 6000Lx, temperature 26 ~ 30 DEG C of condition lower refining seedlings 15 ~ 20 days by silvery birch;
7) transplant and manage: by the Nursery stock transplanting after hardening in transplanting medium, carrying out field planting management.
Further, step 1) in time of explant collection be 6 ~ September, gather 25 ~ 30cm part of the new coppice shoot top tip, and cut off blade.
Further, step 2) in the explant concrete grammar of disinfecting be: the branch of collection is cleaned up, cuts into that 3 ~ 4cm is long, the stem section of band 1 ~ 2 axil, with the HgCl of 0.08 ~ 0.12%v/v in gnotobasis
2soaking disinfection 3 ~ 7min, period constantly shakes and guarantees that thimerosal fully contacts with branch, then clean by sterile water wash, finally stem section two ends is respectively cut 0.4 ~ 0.6cm, leaves the middle axil stem section that contains as aseptic explant.
Further, step 3) in the formula of clump bud inducement medium be: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.48 ~ 0.52mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
Further, step 4) in the formula of subculture multiplication medium be: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.2 ~ 0.3mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
Further, step 5) in the formula of root induction medium be: 412 ~ 413mg/LNH
4nO
3, 473 ~ 477mg/LKNO
3, 42 ~ 43mg/LKH
2pO
4, 92 ~ 93mg/LMgSO
47HO
2, 108 ~ 112mg/LCaCl
22HO
2, 11 ~ 11.5mg/LMnSO
44HO
2, 4 ~ 4.5mg/LZnSO
47HO
2, 2.8 ~ 3.4mg/LH
3bO
3, 0.12 ~ 0.13mg/LNa
2moO
42HO
2, 13.5 ~ 14.5mg/LFeSO
47HO
2, 18.3 ~ 18.9mg/LNa
2eDTA2HO
2, 0.41 ~ 0.42mg/LKI, 0.012 ~ 0.013mg/LCuSO
45HO
2, 0.012 ~ 0.013mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.45 ~ 0.55mg/L puridoxine hydrochloride, 0.45 ~ 0.55mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.65 ~ 0.75mg/LNAA, 18 ~ 22g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
Further, step 3) and step 4) in the condition of culture cultivated of clump bud inducement and shoot proliferation be: temperature 23 ~ 27 DEG C, illumination 9 ~ 11hd
-1, intensity of illumination 2200 ~ 2700Lx; Step 5) in root induction cultivate condition of culture be: temperature 24 ~ 26 DEG C, illumination 8 ~ 10hd
-1, intensity of illumination 2200 ~ 2700Lx.
Further, step 7) described in transplant medium and contain 45 ~ 55% yellow mud and 45 ~ 55% peat soil matrix.
Further, step 7) described in the concrete grammar of field planting management be: is first 90 ~ 95% in sunshade rate after transplanting, relative moisture is 85% ~ 90%, and temperature is cultivate 7 ~ 8 days under the condition of 25 ~ 33 DEG C; Then nursery stock being placed in sunshade rate is 75% ~ 80%, and humidity is 75% ~ 80%, and temperature is cultivate 8 ~ 20 days under the condition of 25 ~ 33 DEG C; Sunshade and trickle management are carried out in nursery routinely later.
Further, step 7) in transplant time be 3 ~ June or 9 ~ November.
The invention has the beneficial effects as follows:
The present invention establishes and a kind ofly can obtain in large quantities by the method for tissue culture of silvery birch clone nursery stock, wherein, the present invention improves ground by the clump bud inducement medium of silvery birch, proliferated culture medium and root media by research, especially the selection to the hormone kind in these three kinds of medium, concentration is embodied in, the improvement of trace and macroelement concentration, just ensure that by the high yield of silvery birch clone nursery stock, growth is fast, and the desired result of nursery stock stalwartness.
It is high by the productive rate of silvery birch clone nursery stock that the inventive method obtains ground, and the time is fast, and the nursery stock obtained is healthy and strong, and these nursery stocks are with solving growing industry by problem that silvery birch nursery stock lacks.
The inventive method obtains the ground of a large amount of high-quality health by silvery birch nursery stock by this vegetative method of tissue cultures, and this vegetative method can guarantee that nursery stock is inherited completely and keeps all fine quality of original maternal plant.
Accompanying drawing explanation
Fig. 1 is over the ground by impact that silvery birch plantlet in vitro is taken root containing the root media of variable concentrations macroelement;
Fig. 2 is over the ground by impact that silvery birch plantlet in vitro is taken root containing the root media of variable concentrations trace element;
Fig. 3 is that different N AA concentration (mg/L) is over the ground by impact that silvery birch plantlet in vitro is taken root.
Embodiment
By a method for silvery birch tissue cultures, comprise the following steps:
1) collection of explant;
2) sterilization of explant;
3) clump bud inducement is cultivated: be seeded in clump bud inducement medium by the explant after upper step sterilization, Fiber differentiation 3 ~ 5 months, period need be forwarded to new clump bud inducement medium 3 ~ 5 times, and explant can be induced out multiple clumps of buds;
4) shoot proliferation is cultivated: the ground upper one-step inducing being gone out clump bud in subculture multiplication medium by silvery birch tissue culture plant inoculation, is often cultivated to be forwarded in new subculture multiplication medium by silvery birch propagation seedling on ground for 28 ~ 32 days and continued to cultivate;
5) root induction is cultivated: cut by the simple bud highly reaching more than 2cm in Multiplying culture in access root media, cultivate 10 ~ 15 days;
6) hardening: enter the hardening stage after the propagation seedling when at least 60% starts to take root, is taken root the ground of acquisition for plantlet in vitro light intensity 3000 ~ 6000Lx, temperature 26 ~ 30 DEG C of condition lower refining seedlings 15 ~ 20 days by silvery birch;
7) transplant and manage: by the Nursery stock transplanting after hardening in transplanting medium, carrying out field planting management.
Preferably, step 1) in time of explant collection be 6 ~ September, gather 25 ~ 30cm part of the new coppice shoot top tip, and cut off blade.
Preferred, step 1) in time of explant collection be that in 6 ~ 9 months, weather continues point ~ 10 point in the morning 9 after sunny 4 ~ 6 days.
Preferably, step 2) in the explant concrete grammar of disinfecting be: the branch of collection is cleaned up, cuts into that 3 ~ 4cm is long, the stem section of band 1 ~ 2 axil, with the HgCl of 0.08 ~ 0.12%v/v in gnotobasis
2soaking disinfection 3 ~ 7min, period constantly shakes and guarantees that thimerosal fully contacts with branch, then clean by sterile water wash, finally stem section two ends is respectively cut 0.4 ~ 0.6cm, leaves the middle axil stem section that contains as aseptic explant.
Preferably, step 3) in the formula of clump bud inducement medium be: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.48 ~ 0.52mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
Preferably, step 3) in the condition of culture of clump bud inducement be: temperature 23 ~ 27 DEG C, illumination 9 ~ 11hd
-1, intensity of illumination 2200 ~ 2700Lx.
Preferably, step 4) in the formula of subculture multiplication medium be: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/Lmg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.2 ~ 0.3mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
Preferably, step 4) in the condition of culture of subculture Multiplying culture be: temperature 23 ~ 27 DEG C, illumination 9 ~ 11hd
-1, intensity of illumination 2200 ~ 2700Lx.
Preferably, step 5) in the formula of root induction medium be: 412 ~ 413mg/LNH
4nO
3, 473 ~ 477mg/LKNO
3, 42 ~ 43mg/LKH
2pO
4, 92 ~ 93mg/LMgSO
47HO
2, 108 ~ 112mg/LCaCl
22HO
2, 11 ~ 11.5mg/LMnSO
44HO
2, 4 ~ 4.5mg/LZnSO
47HO
2, 2.8 ~ 3.4mg/LH
3bO
3, 0.12 ~ 0.13mg/LNa
2moO
42HO
2, 13.5 ~ 14.5mg/LFeSO
47HO
2, 18.3 ~ 18.9mg/LNa
2eDTA2HO
2, 0.41 ~ 0.42mg/LKI, 0.012 ~ 0.013mg/LCuSO
45HO
2, 0.012 ~ 0.013mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.45 ~ 0.55mg/L puridoxine hydrochloride, 0.45 ~ 0.55mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.65 ~ 0.75mg/LNAA, 18 ~ 22g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
Preferably, step 5) in root induction cultivate condition of culture be: temperature 24 ~ 26 DEG C, illumination 8 ~ 10hd
-1, intensity of illumination 2200 ~ 2700Lx.
Preferably, step 7) described in transplant medium and contain 45 ~ 55% yellow mud and 45 ~ 55% peat soil matrix.
Preferably, step 7) described in the concrete grammar of field planting management be: is first 90 ~ 95% in sunshade rate after transplanting, relative moisture is 85% ~ 90%, and temperature is cultivate 7 ~ 8 days under the condition of 25 ~ 33 DEG C; Then nursery stock being placed in sunshade rate is 75% ~ 80%, and humidity is 75% ~ 80%, and temperature is cultivate 8 ~ 20 days under the condition of 25 ~ 33 DEG C; Sunshade and trickle management are carried out in nursery routinely later.
Preferably, step 7) in transplant time be 3 ~ June or 9 ~ November.
Preferred, step 7) in time of transplanting be 3 ~ April.
Preferably, before transplantation, transplant medium with 1/1000 disinfecting solution of potassium permanganate, transplant after each week with 1/800 tpn and carbendazim spray in turn, prevent miscellaneous bacteria from growing, transplant and within latter 15 days, start fertilising, adopt composite fertilizer's solution of 1/1000 to spray weekly once.
Below in conjunction with specific embodiment, the present invention is further illustrated, but be not limited thereto.
Embodiment 1 ground is by the method for silvery birch tissue cultures
Ground is as follows by silvery birch tissue cultures concrete steps:
(1) collection of explant
In 6 ~ September that rainfall is less, after weather continues sunny 5 days, point ~ 10 point, gathered the top tip 25 ~ 30cm of new coppice shoot, cut off blade and put into clean freshness protection package, and take back laboratory disinfection in time the morning 9.
(2) sterilization of explant
Soak branch 30min with washing powder solution, then rinse well at running water down-flow water.Branch is cut into the stem section that 3 ~ 4cm is long, be with 1 ~ 2 axil, with the HgCl of 0.1% on superclean bench
2soaking disinfection 3 ~ 7min, period constantly shakes and guarantees that thimerosal fully contacts with branch, then uses sterile water wash 4 times, each 5min, finally stem section two ends are respectively cut 0.5cm, leave the middle axil stem section that contains as aseptic explant, be inoculated on clump bud inducement medium.
(3) clump bud inducement is cultivated
Explant gathers and after sterilizing, first induced synthesis clump bud on clump bud inducement medium, induction time is 3 ~ 5 months, and midway needs switching and changes fresh culture 3 ~ 5 times, the clump bud induction rate about 15% of explant.Through experimental studies have found that, ground is needed higher 6-BA concentration by during silvery birch clump bud inducement.The condition of culture of clump bud inducement is: temperature 23 ~ 27 DEG C, illumination 9 ~ 11hd
-1, intensity of illumination 2200 ~ 2700Lx.
The formula of above-mentioned clump bud inducement medium is: 412.5mg/LNH
4nO
3, 1900mg/LKNO
3, 85mg/LKH
2pO
4, 370mg/LMgSO
47HO
2, 132mg/LCaCl
22HO
2, 22.3mg/LMnSO
44HO
2, 8.6mg/LZnSO
47HO
2, 6.2mg/LH
3bO
3, 0.25mg/LNa
2moO
42HO
2, 27.8mg/LFeSO
47HO
2, 37.3mg/LNa
2eDTA2HO
2, 0.83mg/LKI, 0.025mg/LCuSO
45HO
2, 0.025mg/LCoCl
26HO
2, 2mg/L glycine, 0.1mg/L thiamine hydrochloride, 0.5mg/L puridoxine hydrochloride, 0.5mg/L nicotinic acid, 100mg/L inositol, 1g/LPVP, 0.5mg/L6-BA, 0.05mg/LIBA, 30g/L sucrose and 7 ~ 8g/L carragheen, pH5.8 (see table 2).
(4) shoot proliferation is cultivated
The ground upper one-step inducing being gone out clump bud by silvery birch plantlet in vitro be transferred to subculture multiplication medium carries out breed and expand numerous.The condition of culture of shoot proliferation is: temperature is 25 ± 2 DEG C, illumination 10h/d, intensity of illumination 2500Lx, and switching in every 30 days once, is cultivated 3 ~ 5 months.In order to configure compatibly by the subculture multiplication medium that silvery birch plantlet in vitro grows, the present embodiment conducts in-depth research Relationship with other Nutrient Elements.
1. macroelement
By the comparative trial of macroelement, the NH in macroelement
4nO
3, KH
2pO
4, CaCl
2the key factor of impact propagation seedling state, containing 0.25MSNH in medium
4nO
3+ 0.5MSKH
2pO
4+ 0.3MSCaCl
22HO
2+ 1MSKNO
3+ 1MSMgSO
47HO
2can make group train propagation seedling keep excellent condition, otherwise propagation seedling there will be top withered fall situation.
2. hormone
Find that the concentration of 6-BA is the deciding factor determining plantlet in vitro proliferation times, height and vitrifying degree by research.
In order to study hormone concentration over the ground by the impact of silvery birch propagation seedling, 0.2mg/L, 0.3mg/L, 0.4mg/L, 0.5mg/L4 concentration gradient is devised to the hormone 6-BA concentration in proliferated culture medium, each concentration processes 10 bottles at every turn, every bottle graft kind propagation bud 4, cultivate after 30 days, statistics proliferation results.
Table 1 hormone concentration is over the ground by the impact of silvery birch propagation seedling
Research shows that the concentration of 6-BA has remarkable impact (see table 1) to proliferation times, the height of breeding seedling, the vitrified degree of propagation seedling.Although when 6-BA concentration is 0.5mg/L, proliferation times can reach 2 times, the height of propagation seedling is too low, is unfavorable for the root induction in later stage, and the vitrified ratio of propagation seedling is also higher, and the ratio of healthy seedling can be caused to decline.Consider the factor of each side, the concentration of 6-BA at 0.2 ~ 0.3mg/L, be best suited for by the Multiplying culture of silvery birch plantlet in vitro.
In sum, need the concentration (table 1) suitably reducing 6-BA when shoot proliferation is cultivated, the final formula obtaining best subculture multiplication medium is: containing the modified MS medium of 0.2 ~ 0.3mg/L6-BA and 0.05mg/LIBA;
Subculture multiplication medium is specifically filled a prescription and is: 412.5mg/LNH
4nO
3, 1900mg/LKNO
3, 85mg/LKH
2pO
4, 370mg/LMgSO
47HO
2, 132mg/LCaCl
22HO
2, 22.3mg/LMnSO
44HO
2, 8.6mg/LZnSO
47HO
2, 6.2mg/LH
3bO
3, 0.25mg/LNa
2moO
42HO
2, 27.8mg/LFeSO
47HO
2, 37.3mg/LNa
2eDTA2HO
2, 0.83mg/LKI, 0.025mg/LCuSO
45HO
2, 0.025mg/LCoCl
26HO
2, 2mg/L glycine, 0.1mg/L thiamine hydrochloride, 0.5mg/L puridoxine hydrochloride, 0.5mg/L nicotinic acid, 100mg/L inositol, 1g/LPVP, 0.2 ~ 0.3mg/L6-BA, 0.05mg/LIBA, 30g/L sucrose, 7 ~ 8g/L carragheen, pH5.8 (see table 2).
(5) root induction
When breeding seedling and transferring each time, the ground highly reaching more than 2cm is cut access root induction medium by silvery birch group training simple bud, cultivate 10 ~ 15 days, cultivation temperature 24 ~ 26 DEG C, illumination 8 ~ 10hd in Multiplying culture
-1, intensity of illumination 2500lx.In order to configure the root induction medium of compatibly being taken root by silvery birch plantlet in vitro, the present embodiment conducts in-depth research Relationship with other Nutrient Elements.
By studying the content of macroelement, trace element, hormone in root media, find when macroelement content be 0.25MS, micronutrient levels be 0.5MS, NAA concentration be 0.7mg/L time, the most applicablely taken root (see Fig. 1, Fig. 2, Fig. 3) by silvery birch plantlet in vitro inductively.
According to the experimental result of Fig. 1 ~ 3, the most applicablely by the prescription of rooting medium that silvery birch plantlet in vitro is taken root be inductively: 412.5mg/LNH
4nO
3, 475mg/LKNO
3, 42.5mg/LKH
2pO
4, 92.5mg/LMgSO
47HO
2, 110mg/LCaCl
22HO
2, 11.15mg/LMnSO
44HO
2, 4.3mg/LZnSO
47HO
2, 3.1mg/LH
3bO
3, 0.125mg/LNa
2moO
42HO
2, 13.9mg/LFeSO
47HO
2, 18.65mg/LNa
2eDTA2HO
2, 0.415mg/LKI, 0.0125mg/LCuSO
45HO
2, 0.0125mg/LCoCl
26HO
2, 2mg/L glycine, 0.1mg/L thiamine hydrochloride, 0.5mg/L puridoxine hydrochloride, 0.5mg/L nicotinic acid, 100mg/L inositol, 0.7mg/LNAA, 20g/L sucrose, 7 ~ 8g/L carragheen, pH5.8 (see table 2).
Table 2 ground is by the culture medium prescription of silvery birch tissue-culturing rapid propagation
Note: the usage amount of carragheen can adjust according to the product of different manufacturers.
(6) hardening
The hardening stage can be proceeded to after propagation seedling when at least 60% starts to take root.Be: intensity of illumination is 6000 ~ 8000lx, temperature 26 ~ 30 DEG C that the suitable hardening time is 15 ~ 20 days aptly by silvery birch training tissue culture seedling condition of taking root.Through hardening, height of seedling of taking root can reach 4 ~ 5cm, well developed root system, and nursery stock is sturdy, leaf color jade green, can directly transplanting in the nutritious bag in nursery.
(7) transplant and manage
Carefully taken out from blake bottle by the offspring wood that has completing hardening, with the medium of the clean nursery stock base portion of clear water rinsing, being implanted into the same day containing mass percent is in the nutritious bag of 50% yellow mud and 50% peat soil matrix, and nutritious bag specification is 6 × 11cm.
Ground is 90 ~ 95% by silvery birch plantlet in vitro transplant early in sunshade rate, and relative moisture is 85% ~ 90%, and temperature is cultivate 7 ~ 8 days under the condition of 25 ~ 33 DEG C; Then nursery stock being placed in sunshade rate is 75% ~ 80%, and humidity is 75% ~ 80%, and temperature is cultivate 8 ~ 20 days under the condition of 25 ~ 33 DEG C; Sunshade and trickle management are carried out in nursery routinely later, and Nursery stock transplanting survival rate is greater than 85%.
Concrete operations are as follows:
1) Nursery stock transplanting 1st ~ 7 days: film is added a cover in seedbed, it is the shading screen of 95% that film is added a cover 1 layer of sunshade rate; Fine day is sprayed 5 ~ 6 times every day, and every day at cloudy day sprays 4 ~ 5 times, sprays every day rainy day 2 ~ 3 times; During temperature height, shading screen drenches temperature lowering water, controlling seedling bed temperature is 25 ~ 33 DEG C.
2) Nursery stock transplanting 8th ~ 30 days: on seedbed, the film of two and side arranges a ventilating opening at interval of 1 meter, lid sunshade on daytime rate is the sunshade net of 85%.During fine day, every day trickle 4 ~ 5 times; Every day at cloudy day trickle 3 ~ 4 times; If the rainy day, then whole day only lid film, every day trickle 0 ~ 1 time.
3) Nursery stock transplanting 31 ~ 60 days: without the need to lid film again, but at the sunshade net still needing lid 50% daytime of fine day, every day trickle 2 ~ 3 times; Cloudy day does not need lid sunshade net, every day trickle 1 ~ 2 time; The little rainy day, do not need lid sunshade net, do not need trickle yet; The large rainy day needs lid film, but need arrange ventilating opening at the two in seedbed and side, does not need lid sunshade net, does not also need trickle.
Before Nursery stock transplanting, transplant medium with 1/1000 disinfecting solution of potassium permanganate, transplant after each week with 1/800 tpn and carbendazim spray in turn, prevent miscellaneous bacteria from growing.Transplant and within latter 15 days, start fertilising, adopt composite fertilizer's solution of 1/1000 to spray once for every 1 week.
At 3 ~ June in Guangdong or 9 ~ 11 monthly portables by silvery birch plantlet in vitro, wherein 3 ~ April be by the best seasonal migration of silvery birch plantlet in vitro, and temperature, humidity are all applicable to the further cultivation of nursery stock, reach afforestation height and requirement.In 7 ~ August, temperature is too high, and transplanting success is on the low side, is applied fertilizer by intensity, and nursery stock can reach afforestation height.10 ~ November, transplanting survival was also higher, but seedling growth mistake in season, and nursery stock there is no method and reached afforestation height spring next year.
The ground originating in Australia owing to having using value mostly at present is artificial culture or natural Hybrid by silvery birch, most self-sterility, and therefore, ground is difficult to obtain by the seed of silvery birch; Seed growing is also difficult to the good characteristic keeping hybrid new breed on the other hand, after introducing a small amount of maternal plant, only has by this pierre technology breeding nursery stock of group training, just can apply these new varieties.It is high by the productive rate of silvery birch clone nursery stock that the inventive method obtains ground, and the time is fast, and the nursery stock obtained is healthy and strong, and these nursery stocks are with solving plant husbandry by problem that silvery birch nursery stock lacks.
Embodiment 2 one kinds of ground are by the method for silvery birch tissue cultures
1) collection of explant: weather continues point ~ 10 point in the morning 9 after sunny 6 days in annual 6 ~ 9 months, gathers 25 ~ 30cm part of the new coppice shoot top tip, and cuts off blade;
2) sterilization of explant: cleaned up by the branch of collection, cuts into the stem section that 3 ~ 4cm is long, be with 1 ~ 2 axil, with the HgCl of 0.08 ~ 0.12%v/v in gnotobasis
2soaking disinfection 3 ~ 7min, period constantly shakes and guarantees that thimerosal fully contacts with branch, then clean by sterile water wash, finally stem section two ends is respectively cut 0.4 ~ 0.6cm, leaves the middle axil stem section that contains as aseptic explant.
3) clump bud inducement is cultivated: be seeded in clump bud inducement medium by the explant after upper step sterilization, Fiber differentiation 4 months, period need be forwarded to new clump bud inducement medium 4 times, and often cultivate and within one month, change a subculture, explant can be induced out multiple clumps of buds; Condition of culture is: temperature 25 DEG C, illumination 10hd
-1, intensity of illumination 2500Lx;
The formula of above-mentioned clump bud inducement medium is: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.48 ~ 0.52mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
4) shoot proliferation is cultivated: the ground upper one-step inducing being gone out clump bud in subculture multiplication medium by silvery birch tissue culture plant inoculation, is often cultivated to be forwarded in new subculture multiplication medium by silvery birch propagation seedling on ground for 28 ~ 32 days and continued to cultivate; Condition of culture is: temperature 25 DEG C, illumination 10hd
-1, intensity of illumination 2500Lx;
The formula of above-mentioned subculture multiplication medium is: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.2 ~ 0.3mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
5) root induction is cultivated: cut by the simple bud highly reaching more than 2cm in Multiplying culture in access root media, cultivate 10 ~ 15 days; Condition of culture is: temperature 245 DEG C, illumination 9hd
-1, intensity of illumination 2700Lx.
The formula of above-mentioned root induction medium is: 412 ~ 413mg/LNH
4nO
3, 473 ~ 477mg/LKNO
3, 42 ~ 43mg/LKH
2pO
4, 92 ~ 93mg/LMgSO
47HO
2, 108 ~ 112mg/LCaCl
22HO
2, 11 ~ 11.5mg/LMnSO
44HO
2, 4 ~ 4.5mg/LZnSO
47HO
2, 2.8 ~ 3.4mg/LH
3bO
3, 0.12 ~ 0.13mg/LNa
2moO
42HO
2, 13.5 ~ 14.5mg/LFeSO
47HO
2, 18.3 ~ 18.9mg/LNa
2eDTA2HO
2, 0.41 ~ 0.42mg/LKI, 0.012 ~ 0.013mg/LCuSO
45HO
2, 0.012 ~ 0.013mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.45 ~ 0.55mg/L puridoxine hydrochloride, 0.45 ~ 0.55mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.65 ~ 0.75mg/LNAA, 18 ~ 22g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
6) hardening: enter the hardening stage after the propagation seedling when at least 60% starts to take root, is taken root the ground of acquisition for plantlet in vitro light intensity 3000 ~ 6000Lx, temperature 30 DEG C of condition lower refining seedlings 15 days by silvery birch;
7) transplant and manage: by the Nursery stock transplanting after hardening in the transplanting medium containing 45 ~ 55% yellow mud and 45 ~ 55% peat soil matrix, carrying out field planting management.The time of transplanting is 3 ~ April.Before transplantation, transplant medium with 1/1000 disinfecting solution of potassium permanganate, transplant after each week with 1/800 tpn and carbendazim spray in turn, prevent miscellaneous bacteria from growing, transplant within latter 15 days, start apply fertilizer, employing 1/1000 composite fertilizer's solution spray weekly once.
The concrete grammar of field planting management is: first after transplanting is 90 ~ 95% in sunshade rate, and relative moisture is 85% ~ 90%, and temperature is cultivate 7 ~ 8 days under the condition of 25 ~ 33 DEG C; Then nursery stock being placed in sunshade rate is 75% ~ 80%, and humidity is 75% ~ 80%, and temperature is cultivate 8 ~ 20 days under the condition of 25 ~ 33 DEG C; Sunshade and trickle management are carried out in nursery routinely later.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from Spirit Essence of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. ground is by a method for silvery birch tissue cultures, it is characterized in that: comprise the following steps:
1) collection of explant;
2) sterilization of explant;
3) clump bud inducement is cultivated: be seeded in clump bud inducement medium by the explant after upper step sterilization, Fiber differentiation 3 ~ 5 months, period need be forwarded to new clump bud inducement medium 3 ~ 5 times, and explant can be induced out multiple clumps of buds;
4) shoot proliferation is cultivated: the ground upper one-step inducing being gone out clump bud in subculture multiplication medium by silvery birch tissue culture plant inoculation, is often cultivated to be forwarded in new subculture multiplication medium by silvery birch propagation seedling on ground for 28 ~ 32 days and continued to cultivate;
5) root induction is cultivated: cut by the simple bud highly reaching more than 2cm in Multiplying culture in access root media, cultivate 10 ~ 15 days;
6) hardening: enter the hardening stage after the propagation seedling when at least 60% starts to take root, is taken root the ground of acquisition for plantlet in vitro light intensity 3000 ~ 6000Lx, temperature 26 ~ 30 DEG C of condition lower refining seedlings 15 ~ 20 days by silvery birch;
7) transplant and manage: by the Nursery stock transplanting after hardening in transplanting medium, carrying out field planting management.
2. method according to claim 1, is characterized in that: the time of explant collection in step 1) is 6 ~ September, gathers 25 ~ 30cm part of the new coppice shoot top tip, and cuts off blade.
3. method according to claim 1, it is characterized in that: step 2) in the explant concrete grammar of disinfecting be: the branch of collection is cleaned up, cut into the stem section that 3 ~ 4cm is long, be with 1 ~ 2 axil, with the HgCl of 0.08 ~ 0.12%v/v in gnotobasis
2soaking disinfection 3 ~ 7min, period constantly shakes and guarantees that thimerosal fully contacts with branch, then clean by sterile water wash, finally stem section two ends is respectively cut 0.4 ~ 0.6cm, leaves the middle axil stem section that contains as aseptic explant.
4. according to method according to claim 1, it is characterized in that: in step 3), the formula of clump bud inducement medium is: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.48 ~ 0.52mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
5. method according to claim 1, is characterized in that: in step 4), the formula of subculture multiplication medium is: 412 ~ 413mg/LNH
4nO
3, 1898 ~ 1902mg/LKNO
3, 84.5 ~ 85.5mg/LKH
2pO
4, 369 ~ 371mg/LMgSO
47HO
2, 131 ~ 133mg/LCaCl
22HO
2, 22 ~ 22.5mg/LMnSO
44HO
2, 8.4 ~ 8.8mg/LZnSO
47HO
2, 6 ~ 6.4mg/LH
3bO
3, 0.22 ~ 0.27mg/LNa
2moO
42HO
2, 27.5 ~ 28mg/LFeSO
47HO
2, 37 ~ 37.5mg/LNa
2eDTA2HO
2, 0.8 ~ 0.85mg/LKI, 0.023 ~ 0.027mg/LCuSO
45HO
2, 0.023 ~ 0.027mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.48 ~ 0.52mg/L puridoxine hydrochloride, 0.48 ~ 0.52mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.8 ~ 1.2g/LPVP, 0.2 ~ 0.3mg/L6-BA, 0.048 ~ 0.052mg/LIBA, 27 ~ 32g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
6. according to method according to claim 1, it is characterized in that: in step 5), the formula of root induction medium is: 412 ~ 413mg/LNH
4nO
3, 473 ~ 477mg/LKNO
3, 42 ~ 43mg/LKH
2pO
4, 92 ~ 93mg/LMgSO
47HO
2, 108 ~ 112mg/LCaCl
22HO
2, 11 ~ 11.5mg/LMnSO
44HO
2, 4 ~ 4.5mg/LZnSO
47HO
2, 2.8 ~ 3.4mg/LH
3bO
3, 0.12 ~ 0.13mg/LNa
2moO
42HO
2, 13.5 ~ 14.5mg/LFeSO
47HO
2, 18.3 ~ 18.9mg/LNa
2eDTA2HO
2, 0.41 ~ 0.42mg/LKI, 0.012 ~ 0.013mg/LCuSO
45HO
2, 0.012 ~ 0.013mg/LCoCl
26HO
2, 1.8 ~ 2.2mg/L glycine, 0.08 ~ 0.12mg/L thiamine hydrochloride, 0.45 ~ 0.55mg/L puridoxine hydrochloride, 0.45 ~ 0.55mg/L nicotinic acid, 99 ~ 101mg/L inositol, 0.65 ~ 0.75mg/LNAA, 18 ~ 22g/L sucrose, 7 ~ 8g/L carragheen, pH5.75 ~ 5.85.
7. according to method according to claim 1, it is characterized in that: the condition of culture that in step 3) and step 4), clump bud inducement and shoot proliferation are cultivated is: temperature 23 ~ 27 DEG C, illumination 9 ~ 11hd
-1, intensity of illumination 2200 ~ 2700Lx; The condition of culture that in step 5), root induction is cultivated is: temperature 24 ~ 26 DEG C, illumination 8 ~ 10hd
-1, intensity of illumination 2200 ~ 2700Lx.
8. method according to claim 1, is characterized in that: transplant medium described in step 7) and contain 45 ~ 55% yellow mud and 45 ~ 55% peat soil matrix.
9. method according to claim 1, is characterized in that: the concrete grammar of field planting management described in step 7) is: first after transplanting is 90 ~ 95% in sunshade rate, and relative moisture is 85% ~ 90%, and temperature is cultivate 7 ~ 8 days under the condition of 25 ~ 33 DEG C; Then nursery stock being placed in sunshade rate is 75% ~ 80%, and humidity is 75% ~ 80%, and temperature is cultivate 8 ~ 20 days under the condition of 25 ~ 33 DEG C; Sunshade and trickle management are carried out in nursery routinely later.
10. method according to claim 1, is characterized in that: the time of transplanting in step 7) is 3 ~ June or 9 ~ November.
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| CN107926675A (en) * | 2017-12-13 | 2018-04-20 | 佛山推启农业研究院（普通合伙） | A kind of implantation methods of selenium-rich water spinach |
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Non-Patent Citations (6)
| Title |
|---|
| 张晓明等: "莫丽银桦组培技术研究", 《林业科技开发》 * |
| 李湘阳等: ""罗宾戈登"银桦组培苗不定根的诱导", 《北方园艺》 * |
| 萧洪东等: "红花银桦的组织培养与快速繁殖", 《植物生理学通讯》 * |
| 赵平丽等: "白玉银桦组培快繁技术研究", 《安徽农业科学》 * |
| 赵平丽等: "白翅银桦组培快繁技术研究", 《江西林业科技》 * |
| 龚峥等: "绵毛银桦的组织培养与快速繁殖", 《广东农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107926675A (en) * | 2017-12-13 | 2018-04-20 | 佛山推启农业研究院（普通合伙） | A kind of implantation methods of selenium-rich water spinach |
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