CN102577952B - Tissue culturing method for quercus virginiana - Google Patents

Tissue culturing method for quercus virginiana Download PDF

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CN102577952B
CN102577952B CN2012100297273A CN201210029727A CN102577952B CN 102577952 B CN102577952 B CN 102577952B CN 2012100297273 A CN2012100297273 A CN 2012100297273A CN 201210029727 A CN201210029727 A CN 201210029727A CN 102577952 B CN102577952 B CN 102577952B
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culture
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explant
tissue culturing
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CN102577952A (en
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王树凤
孙海菁
陈益泰
路晓宏
陈雨春
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Shangyu Century Sunshine Gardens Engineering Co Ltd
TAIDONG TONGYUAN SEEDING FIELD
Research Institute of Subtropical Forestry of Chinese Academy of Forestry
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Shangyu Century Sunshine Gardens Engineering Co Ltd
TAIDONG TONGYUAN SEEDING FIELD
Research Institute of Subtropical Forestry of Chinese Academy of Forestry
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Abstract

A tissue culturing method for quercus virginiana belongs to plant tissue culturing methods and is characterized by comprising the following steps: 1 selecting of explants; 2 disinfection of the explants; 3 primary culture; 4 multiplication culturing of adventitious buds; 5 culturing of strong seedlings; 6 culturing of strike roots; and 7 seedling acclimatization and transplanting. The tissue culturing method for the quercus virginiana can keep stress resistance and enjoyment of a female parent and can effectively increase propagation coefficient of the quercus virginiana. Compared with seminal propagation, tissue culturing reduces dependency on imported seeds, can effectively reduce nursery stock cost, and saves expenditures. Compared with cutting propagation, the tissue culturing is short in seedling culturing period, can obtain the nursery stock with strike roots within 4 months, and saves cost.

Description

A kind of tissue culturing method for quercus virginiana
Technical field
The invention belongs to method for plant tissue culture, be specially a kind of tissue culturing method for quercus virginiana.
Background technology
Virginia oak ( Quercus virginiana)Belong to Fagaceae (Fagaceae) oak and belong to (Quercus) seeds, originate in the U.S., be the high megaphanerophyte of evergreen broad-leaved.Single leaf, alternate, the Lao Ye keratin, leaf surface bottle green tool gloss, the green whiting in the back side, the proterties such as leaf color, shape, size, quality have very strong plasticity with impacts such as developmental stage, environment, the large and meat of young leaves, the color pale green, the leaf margin tooth lacks greatly; Little and the keratin of Lao Ye, the leaf margin tooth is little and many.Bloom early spring, brightly yellowish look, and ripening fruits is brown, is commonly called as acorn, for birds and animal provide abundant food source, namely falls after ripe, and seed vitality is lower.The well-drained sandy soil of Virginia oak happiness can salt spray resistance and higher soil salt, and is responsive to soil moisture, can stand moderate arid, and clay also can be grown, and can tolerate the low temperature of minimum-13 ℃.Virginia oak juvenile growth speed, seedling can grow to 4 feet (1.2m) at First Year, and along with growing up of nursery stock, growth rate can reduce.On average can reach 15m the oak that grows up not of hard-core Growth In Space high, the diameter of a cross-section of a tree trunk 1.3 meters above the ground can reach 91-122cm, and more than the tree crown scope can reach 46m, the life-span reached centuries.Virginia oak mainly is distributed in U.S.'s southeastern coast and Gulf of Mexico seashore, namely from Virginia downstate, the Georgia State, Florida State, Louisiana State, until the middle and south, Texas one line; Breathing out the Ma Zhou west and south and Mexico's Northeast Mountain Areas also has fragmentary distribution at the Russia carat, is the important composition seeds in the coastal hard wealthy woods of the U.S. and shrubbery forest land, is widely used as city shade tree and ornamental trees and shrubs in U.S.'s southeastern coast.
The Virginia oak seed that Inst. of Subtropical Forestry, Chinese Academy of Forestry Sciences introduces from Missouri, USA, after seedling, about 5% plant trunk is obvious, and high growth is outstanding, and branch is sparse, and is crown narrow and small; Most nursery stocks do not have obvious trunk, and branches and leaves are intensive, crown the first transaction of a day's business, or be the how dried state of growing thickly.After live oak introduction, all can grow in subtropics, torrid areas such as China Zhejiang, Shanghai, Jiangsu, Fujian, simultaneously because Virginia oak has very strong Salt And Alkali Tolerance ability, well developed root system in addition, germinating property is strong, branch tool toughness, has the ability of resisting hurricane, therefore be widely used in the coast protection forest construction in China's Yangtze River Delta area, having made up the defective that lacks evergreen broad-leaved wind resistance salt tolerant seeds in present China coast protection forest system, is the evergreen tall and big arbor species of unique a kind of salt tolerant in present southeastern coast protection forest system.Simultaneously, acorn has attracted large quantities of birds and animal to collect, and has enriched the bio-diversity of coast protection forest system.Virginia oak also can be used for the public leisure place greening such as garden, park, farm, and its tree crown is wide, and the leaf look vivid, for people good shade, the habitat of Condom and animal.
Every annual meeting produces a large amount of rubbers fruits to Virginia oak in the original producton location, therefore mostly adopts seminal propagation, only has the clone that minority nursery stock company relies on vegetative propagation to obtain various trait to be used for different breeding objectives.The most dependence on import seminal propagation of present domestic Virginia oak nursery stock, but because the Virginia oak seed does not have resting stage, can sprout as long as temperature is suitable, therefore in import kind subprocess, due to reasons such as mechanical damages, make greatly seed can not get sowing in time after sprouting and devitalization causes emergence rate low, seedling cost is higher.Inst. of Subtropical Forestry, Chinese Academy of Forestry Sciences introduces first at home Virginia oak and carries out relevant research, and carried out Virginia oak vegetative propagation technique research, obtained very big breakthrough (ZL200910097684.0) aspect cuttage propagation method of Virginia oak, survival rate can be up to 80-85%.
Summary of the invention
For the above-mentioned problems in the prior art, the object of the invention is to design provides a kind of technical scheme that significantly improves the tissue culturing method for quercus virginiana of its reproduction coefficient, and greatly shortened the Virginia oak growing-seedling period, solve the mode that the dependence on import seed is grown seedlings, saved cost.
Described a kind of tissue culturing method for quercus virginiana is characterized in that comprising the following steps:
1) choosing of explant: explant is taken from the seedling of land for growing field crops plant or indoor cultivation;
2) explant sterilization: get stem with bud as explant, explant is after running water rinses 10-14 hour, then with alcohol disinfecting 25-35 second of 70-75%, then mercuric chloride soaked 3-5 minute, then with aseptic water washing 2-3 time, is placed in aseptic vial standby;
3) first culture: the explant of sterilize was cut in the stem with bud access medium of 0.8-1.2cm cultivation 25-35 days, gave approximately 2500-3500 Lx of illumination, photoperiod 14-18hr/6-10hr, 25 ℃ ± 2 ℃ of temperature; Described just culture base is comprised of WPM medium or 1/4MS medium+6-benzyladenine 0.8-1.2mg/L;
4) adventitious bud proliferation is cultivated: the stem with bud that the growth sign is arranged in first culture base is accessed in proliferated culture medium breed cultivation, the same step 3) of condition of culture, cultivation cycle are 40-50 days; Described proliferated culture medium is comprised of WPM or 1/4MS+6-benzyladenine 0.1-0.3mg/L+ growth regulatory substance methyl α-naphthyl acetate 0.4-0.6 mg/L;
5) strong seedling culture: proliferated culture medium middle period look is dark green, and the indefinite bud of length 1-2cm downcuts one by one, cultivates 25-35 days the same step 3) of condition of culture in access WPM or 1/4MS medium;
6) culture of rootage: bud healthy and strong after strong seedling culture is cut in base portion brown material and callus access root media carry out culture of rootage, first secretly cultivate 1.5-2.5 week, then take root after illumination cultivation 3.5-4.5 week, the same step 3) of condition of culture; Described root media is comprised of WPM or 1/4MS+ growth regulatory substance 6-benzyladenine 0.8-1.2 mg/L+growth regulatory substance methyl α-naphthyl acetate 0.4-0.6 mg/L;
7) hardening and transplanting: will have the medium bottle cap of the Virginia oak seedling that takes root to open, under room temperature, the room temperature lower refining seedling after 2-4 days, takes out seedling, wash away the root medium, transplant and be equipped with on the seedbed of river sand matrix, keep 20-25 ℃ of temperature, humidity 85%-95%, suitably shading gets final product.
Described a kind of tissue culturing method for quercus virginiana is characterized in that in step 1):
The land for growing field crops plant: generally should draw materials in the first half of the year, choosing the stem with bud of sprouting then is explant, and the explant that the land for growing field crops gathers separately adds 1-2 to drip the Tween 80 sterilization;
Indoor cultivation: choose the Virginia oak seed without small holes caused by worms, full grains, soaked 22-26 hour in running water, discard floating seed, then soak with saturated bleaching powder supernatant and carried out surface sterilization in 25-35 minute, planting seed after sterilization is in the plastic containers of 30cm * 20cm * 8cm, cover river sand in container, river sand thickness is 4-6cm, plastic containers are placed in illumination box to be cultivated, 25 ± 1 ℃ of temperature, intensity of illumination 11000-13000Lx, light/dark cycle is 14-18h/6-10h.
Described a kind of tissue culturing method for quercus virginiana is characterized in that adding respectively sucrose 15-25g.L-1 in described first culture base, adventitious bud proliferation medium, strong seedling culture base, root media, and agar 0.5-1.0% regulates PH to 5.5-5.9.
Described a kind of tissue culturing method for quercus virginiana, it is characterized in that to take explant anti-browning measure in the described first culture process of step 3), be specially: just adding the active carbon that adds 0.2-0.4% or 0.4-0.6% in the culture base, or repeatedly switching repeatedly, be after explant is inoculated for the first time, cultivated 2-3 days, and be transferred to same medium, turn 2-3 time like this.
Described a kind of tissue culturing method for quercus virginiana is characterized in that described WPM medium is by being comprised of macroelement, trace element, molysite, organic principle;
Described macroelement is: (1) ammonium nitrate NH 4NO 3, 400 mgL -1, (2) nitrate of lime Ca (NO 3) 2, 556 mgL -1, (3) calcium chloride CaCl 2H 2O, 96mgL -1, (4) magnesium sulfate MgSO 47H 2O, 370 mgL -1, (5) potassium sulphate K 2SO 4, 990 mgL -1, (6) potassium dihydrogen phosphate KH 2PO 4, 170 mg L -1
Described trace element is: (1) boric acid H 3BO 3, 6.2 mgL -1, (2) sodium molybdate Na 2MoO 42H 2O, 0.25 mgL -1, (3) manganese sulphate MnSO 44H 2O, 22.3 mgL -1, (4) copper sulphate CuSO 45H 2O, 0.25 mgL -1, (5) zinc sulphate ZnSO 47H 2O, 8.6 mgL -1
Described molysite is: (1) disodium ethylene diamine tetraacetate Na 2EDTA, 37.3 mgL -1, (2) ferrous sulfate FeSO 47H2O, 27.8 mgL -1
Described organic principle is: (1) inositol C 6H 12O 62H 2O, 100 mgL -1, (2) nicotinic acid NC 5H 4COOH, 0.5 mgL -1, (3) thiamine hydrochloride C 12H 17ClOS2HCl, 1.6 mgL -1
Described a kind of tissue culturing method for quercus virginiana is characterized in that described MS medium is comprised of macroelement, trace element, molysite, organic principle;
Described macroelement is: (1) potassium nitrate KNO 3, 1900 mgL -1, (2) ammonium nitrate NH 4NO 3, 1650 mgL -1, (3) potassium dihydrogen phosphate KH 2PO 4, 170 mg L -1, (4) magnesium sulfate MgSO 47H 2O, 370 mgL -1, (5) calcium chloride CaCl 2H 2O, 440 mgL -1
Described trace element is: (1) potassium iodide KI, 0.83 mgL -1, (2) boric acid H 3BO 3, 6.2 mgL -1, (3) manganese sulphate MnSO 44H 2O, 22.3 mgL -1, (4) zinc sulphate ZnSO 47H 2O, 8.6 mgL -1(5) sodium molybdate Na 2MoO 42H 2O, 0.25 mgL -1(6) copper sulphate CuSO 45H 2O, 0.025 mgL -1, (7) cobalt chloride CoCl 26H 2O, 0.025 mgL -1
Described molysite is: (1) disodium ethylene diamine tetraacetate Na 2EDTA, 37.3 mgL -1, (2) ferrous sulfate FeSO 47H2O, 27.8 mgL -1
Described organic principle is: (1) inositol C 6H 12O 62H 2O, 100 mgL -1(2) glycine NH 2CH 2COOH, 2 mgL -1, (3) thiamine hydrochloride C 12H 17ClOS2HCl, 0.1 mgL -1(4) puridoxine hydrochloride C 8H 11O 3NHCl, 0.5 mgL -1(5) nicotinic acid NC 5H 4COOH, 0.5 mgL -1
Described a kind of tissue culturing method for quercus virginiana is characterized in that 1/4MS medium used is that macroelement is 1/4 of MS minimal medium, and all the other composition trace elements, molysite, organic principle are with the MS minimal medium.
Above-mentioned a kind of tissue culturing method for quercus virginiana can keep maternal resistance and sight preferably, effectively improves the Virginia oak reproduction coefficient.Compare with seminal propagation, tissue is cultivated the dependence that reduces the import seed, can effectively reduce seedling cost, reduces expenses; Compare with cottage propagation, the tissue cultivating and seedling cycle is short, can obtain the nursery stock that takes root, and save cost in 4 months.
Embodiment
The invention will be further described below in conjunction with specific embodiment.
Embodiment 1: a kind of tissue culturing method for quercus virginiana, and its incubation step is as follows:
(1) choosing of explant: explant picks up from the plant of cultivation in the inferior woods degeneration-resistant physiological ecological of institute laboratory, Chinese forest-science academy.
(2) explant sterilization: get stem with bud as explant, explant is after running water rinses 12 hours, again with 30 seconds (if explants that the land for growing field crops gathers of the alcohol disinfecting of 70-75%, add 1-2 to drip Tween 80, to strengthen the infiltration of disinfectant), then mercuric chloride soaked 3-5 minute, then with aseptic water washing for several times, was placed in aseptic vial standby.
(3) first culture: the explant of sterilize was cut in the stem with bud access WPM of 1cm left and right or 1/4MS+ growth regulatory substance 6-benzyladenine 1.0mg/L medium cultivation 30 days, gave approximately 3000lx of illumination, photoperiod 16hr/8hr, 25 ℃ ± 2 ℃ of temperature.Because the Virginia oak explant contains than polyphenols, therefore just need take the anti-browning measure in the culture process, concrete measure comprises: the active carbon that adds 0.2-0.4% or 0.4-0.6% in the medium, or repeatedly switching repeatedly, be after explant is inoculated for the first time, cultivated 3 days, be transferred to same medium, turn like this 3 times, can effectively prevent brownization generation.
(4) adventitious bud proliferation is cultivated: the stem with bud that the growth sign is arranged in first culture base is accessed in WPM or 1/4MS+ growth regulatory substance 6-benzyladenine 0.2mg/L+ growth regulatory substance methyl α-naphthyl acetate 0.5 mg/L medium breed cultivation, condition of culture is with the first culture of step 3), and cultivation cycle is 45 days.
(5) strong seedling culture: proliferated culture medium middle period look is dark green, and the indefinite bud of length 1-2cm downcuts one by one, cultivates 30 days in access WPM or 1/4MS medium, and condition of culture is with the first culture of step 3).
(6) culture of rootage: bud healthy and strong after strong seedling culture is cut in base portion brown material and callus access WPM or 1/4MS+ growth regulatory substance 6-benzyladenine 1.0 mg/L+ growth regulatory substance methyl α-naphthyl acetate 0.5 mg/L medium carry out culture of rootage, first secretly cultivated for 2 weeks, illumination cultivation was taken root after 4 weeks again, and condition of culture is with the first culture of step 3).
(7) hardening and transplanting: will have the medium bottle cap of the Virginia oak seedling that takes root to open, under room temperature, the room temperature lower refining seedling is after 3 days, take out seedling, wash away the root medium, transplanting is equipped with on the seedbed of river sand matrix, keeps 20-25 ℃ of temperature, humidity 85%-95%, suitably shade, survival rate can reach more than 90%.
Described WPM medium is comprised of following material:
Described macroelement is: (1) ammonium nitrate NH 4NO 3, 400 mgL -1, (2) nitrate of lime Ca (NO 3) 2, 556 mgL -1, (3) calcium chloride CaCl 2H 2O, 96mgL -1, (4) magnesium sulfate MgSO 47H 2O, 370 mgL -1, (5) potassium sulphate K 2SO 4, 990 mgL -1, (6) potassium dihydrogen phosphate KH 2PO 4, 170 mg L -1
Described trace element is: (1) boric acid H 3BO 3, 6.2 mgL -1, (2) sodium molybdate Na 2MoO 42H 2O, 0.25 mgL -1, (3) manganese sulphate MnSO 44H 2O, 22.3 mgL -1, (4) copper sulphate CuSO 45H 2O, 0.25 mgL -1, (5) zinc sulphate ZnSO 47H 2O, 8.6 mgL -1
Described molysite is: (1) disodium ethylene diamine tetraacetate Na 2EDTA, 37.3 mgL -1, (2) ferrous sulfate FeSO 47H2O, 27.8 mgL -1
Described organic principle is: (1) inositol C 6H 12O 62H 2O, 100 mgL -1, (2) nicotinic acid NC 5H 4COOH, 0.5 mgL -1, (3) thiamine hydrochloride C 12H 17ClOS2HCl, 1.6 mgL -1
Described MS medium is comprised of following material:
Described macroelement is: (1) potassium nitrate KNO 3, 1900 mgL -1, (2) ammonium nitrate NH 4NO 3, 1650 mgL -1, (3) potassium dihydrogen phosphate KH 2PO 4, 170 mg L -1, (4) magnesium sulfate MgSO 47H 2O, 370 mgL -1, (5) calcium chloride CaCl 2H 2O, 440 mgL -1
Described trace element is: (1) potassium iodide KI, 0.83 mgL -1, (2) boric acid H 3BO 3, 6.2 mgL -1, (3) manganese sulphate MnSO 44H 2O, 22.3 mgL -1, (4) zinc sulphate ZnSO 47H 2O, 8.6 mgL -1(5) sodium molybdate Na 2MoO 42H 2O, 0.25 mgL -1(6) copper sulphate CuSO 45H 2O, 0.025 mgL -1, (7) cobalt chloride CoCl 26H 2O, 0.025 mgL -1
Described molysite is: (1) disodium ethylene diamine tetraacetate Na 2EDTA, 37.3 mgL -1, (2) ferrous sulfate FeSO 47H2O, 27.8 mgL -1
Described organic principle is: (1) inositol C 6H 12O 62H 2O, 100 mgL -1(2) glycine NH 2CH 2COOH, 2 mgL -1, (3) thiamine hydrochloride C 12H 17ClOS2HCl, 0.1 mgL -1(4) puridoxine hydrochloride C 8H 11O 3NHCl, 0.5 mgL -1(5) nicotinic acid NC 5H 4COOH, 0.5 mgL -1
Above-mentioned steps 3) in, explant is cut into the stem with bud of 0.8cm or 1.2cm, cultivated 25 days or 35 days, give illumination approximately 2500 Lx, 2600 Lx or 3500 Lx, photoperiod 14hr/6hr or 18hr/10hr, 6-benzyladenine 0.8mg/L or 1.2mg/L in described just culture base; In step 4), cultivation cycle is 40 days or 50 days; 6-benzyladenine 0.1 or 0.3mg/L in described proliferated culture medium, growth regulatory substance methyl α-naphthyl acetate 0.4 or 0.6 mg/L.In step 5), cultivation cycle is 25 or 35 days; First secretly cultivated for 1.5 week or 2.5 weeks in step 6), then 3.5 week or 4.5 weeks of illumination cultivation, in described root media, the growth regulatory substance 6-benzyladenine is 0.8 or 1.2 mg/L, the growth regulatory substance methyl α-naphthyl acetate is 0.4 or 0.6 mg/L.Other step also can reach beneficial effect of the present invention with embodiment 1.

Claims (6)

1. tissue culturing method for quercus virginiana is characterized in that comprising the following steps:
1) choosing of explant: explant is taken from the seedling of land for growing field crops plant or indoor cultivation;
2) explant sterilization: get stem with bud as explant, explant is after running water rinses 10-14 hour, then with alcohol disinfecting 25-35 second of 70-75%, then mercuric chloride soaked 3-5 minute, then with aseptic water washing 2-3 time, is placed in aseptic vial standby;
3) first culture: the explant of sterilize was cut in the stem with bud access medium of 0.8-1.2cm cultivation 25-35 days, gave illumination 2500-3500 Lx, photoperiod 14-18hr/6-10hr, 25 ℃ ± 2 ℃ of temperature; Described just culture base is comprised of WPM medium+6-benzyladenine 0.8-1.2mg/L or 1/4MS medium+6-benzyladenine 0.8-1.2mg/L;
4) adventitious bud proliferation is cultivated: the stem with bud that the growth sign is arranged in first culture base is accessed in proliferated culture medium breed cultivation, the same step 3) of condition of culture, cultivation cycle are 40-50 days; Described proliferated culture medium is comprised of WPM+6-benzyladenine 0.1-0.3mg/L+ growth regulatory substance methyl α-naphthyl acetate 0.4-0.6 mg/L or 1/4MS+6-benzyladenine 0.1-0.3mg/L+ growth regulatory substance methyl α-naphthyl acetate 0.4-0.6 mg/L;
5) strong seedling culture: proliferated culture medium middle period look is dark green, and the indefinite bud of length 1-2cm downcuts one by one, cultivates 25-35 days the same step 3) of condition of culture in access WPM or 1/4MS medium;
6) culture of rootage: bud healthy and strong after strong seedling culture is cut in base portion brown material and callus access root media carry out culture of rootage, first secretly cultivate 1.5-2.5 week, then take root after illumination cultivation 3.5-4.5 week, the same step 3) of condition of culture; Described root media is comprised of WPM+growth regulatory substance 6-benzyladenine 0.8-1.2 mg/L+growth regulatory substance methyl α-naphthyl acetate 0.4-0.6 mg/L or 1/4MS+ growth regulatory substance 6-benzyladenine 0.8-1.2 mg/L+growth regulatory substance methyl α-naphthyl acetate 0.4-0.6 mg/L;
Add respectively sucrose 15-25g.L-1 in described first culture base, adventitious bud proliferation medium, strong seedling culture base, root media, agar 0.5-1.0% regulates PH to 5.5-5.9;
7) hardening and transplanting: will have the medium bottle cap of the Virginia oak seedling that takes root to open, the room temperature lower refining seedling took out seedling after 2-4 days, wash away the root medium, transplant and be equipped with on the seedbed of river sand matrix, keep 20-25 ℃ of temperature, humidity 85%-95%, suitably shading gets final product.
2. a kind of tissue culturing method for quercus virginiana as claimed in claim 1 is characterized in that in step 1):
The land for growing field crops plant: should draw materials in the first half of the year, choosing the stem with bud of sprouting then is explant, separately adds 1-2 to drip the Tween 80 sterilization;
Indoor cultivation: choose the Virginia oak seed without small holes caused by worms, full grains, soaked 22-26 hour in running water, discard floating seed, then soak with saturated bleaching powder supernatant and carried out surface sterilization in 25-35 minute, planting seed after sterilization is in the plastic containers of 30cm * 20cm * 8cm, cover river sand in container, river sand thickness is 4-6cm, plastic containers are placed in illumination box to be cultivated, 25 ± 1 ℃ of temperature, intensity of illumination 11000-13000Lx, light/dark cycle is 14-18h/6-10h.
3. a kind of tissue culturing method for quercus virginiana as claimed in claim 1, it is characterized in that to take explant anti-browning measure in the described first culture process of step 3), be specially: just adding the active carbon that adds 0.2-0.4% or 0.4-0.6% in the culture base, or repeatedly switching repeatedly, be after explant is inoculated for the first time, cultivated 2-3 days, and be transferred to same medium, turn 2-3 time like this.
4. a kind of tissue culturing method for quercus virginiana as claimed in claim 1, is characterized in that described WPM medium is comprised of macroelement, trace element, molysite, organic principle;
Described macroelement is: (1) ammonium nitrate NH 4NO 3, 400 mgL -1, (2) nitrate of lime Ca (NO 3) 2, 556 mgL -1, (3) calcium chloride CaCl 2H 2O, 96mgL -1, (4) magnesium sulfate MgSO 47H 2O, 370 mgL -1, (5) potassium sulphate K 2SO 4, 990 mgL -1, (6) potassium dihydrogen phosphate KH 2PO 4, 170 mg L -1
Described trace element is: (1) boric acid H 3BO 3, 6.2 mgL -1, (2) sodium molybdate Na 2MoO 42H 2O, 0.25 mgL -1, (3) manganese sulphate MnSO 44H 2O, 22.3 mgL -1, (4) copper sulphate CuSO 45H 2O, 0.25 mgL -1, (5) zinc sulphate ZnSO 47H 2O, 8.6 mgL -1
Described molysite is: (1) disodium ethylene diamine tetraacetate Na 2EDTA, 37.3 mgL -1, (2) ferrous sulfate FeSO 47H2O, 27.8 mgL -1
Described organic principle is: (1) inositol C 6H 12O 62H 2O, 100 mgL -1, (2) nicotinic acid NC 5H 4COOH, 0.5 mgL -1, (3) thiamine hydrochloride C 12H 17ClOS2HCl, 1.6 mgL -1
5. a kind of tissue culturing method for quercus virginiana as claimed in claim 1, is characterized in that described MS medium is comprised of macroelement, trace element, molysite, organic principle;
Described macroelement is: (1) potassium nitrate KNO 3, 1900 mgL -1, (2) ammonium nitrate NH 4NO 3, 1650 mgL -1, (3) potassium dihydrogen phosphate KH 2PO 4, 170 mg L -1, (4) magnesium sulfate MgSO 47H 2O, 370 mgL -1, (5) calcium chloride CaCl 2H 2O, 440 mgL -1
Described trace element is: (1) potassium iodide KI, 0.83 mgL -1, (2) boric acid H 3BO 3, 6.2 mgL -1, (3) manganese sulphate MnSO 44H 2O, 22.3 mgL -1, (4) zinc sulphate ZnSO 47H 2O, 8.6 mgL -1(5) sodium molybdate Na 2MoO 42H 2O, 0.25 mgL -1(6) copper sulphate CuSO 45H 2O, 0.025 mgL -1, (7) cobalt chloride CoCl 26H 2O, 0.025 mgL -1
Described molysite is: (1) disodium ethylene diamine tetraacetate Na 2EDTA, 37.3 mgL -1, (2) ferrous sulfate FeSO 47H2O, 27.8 mgL -1
Described organic principle is: (1) inositol C 6H 12O 62H 2O, 100 mgL -1(2) glycine NH 2CH 2COOH, 2 mgL -1, (3) thiamine hydrochloride C 12H 17ClOS2HCl, 0.1 mgL -1(4) puridoxine hydrochloride C 8H 11O 3NHCl, 0.5 mgL -1(5) nicotinic acid NC 5H 4COOH, 0.5 mgL -1
6. a kind of tissue culturing method for quercus virginiana as claimed in claim 1, is characterized in that 1/4MS medium used is that macroelement is 1/4 of MS minimal medium, and all the other composition trace elements, molysite, organic principle are with the MS minimal medium.
CN2012100297273A 2012-02-10 2012-02-10 Tissue culturing method for quercus virginiana Expired - Fee Related CN102577952B (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103535280B (en) * 2013-10-29 2015-10-28 江苏省林业科学研究院 A kind of water oak tissue culture propagation
CN103548679B (en) * 2013-10-29 2015-02-18 江苏省林业科学研究院 Method for quercus nuttallii somatic embryogenesis
CN103636506B (en) * 2013-12-24 2015-06-10 黑龙江省林业科学研究所 method for performing plant culture by utilizing shepherdia argentea caulicle regenerated plant induction culture medium and
CN104839021A (en) * 2015-05-08 2015-08-19 中国林业科学研究院亚热带林业研究所 Quercus nuttallii tissue culturing method
CN106613948A (en) * 2016-10-17 2017-05-10 柳州玲通科技有限责任公司 An initial culture bud inducing method for lithocarpus carolinae
CN106386496A (en) * 2016-10-17 2017-02-15 柳州玲通科技有限责任公司 Primary culture shoot induction method for lithocarpus elmerrillii
CN106613947A (en) * 2016-10-17 2017-05-10 柳州玲通科技有限责任公司 Primarily-cultured bud inducing method of Lithocarpus craibianus
CN107047297A (en) * 2017-02-14 2017-08-18 唐春艳 A kind of rice sweet oak tissue-cultured seedling rooting induction method
CN108243817B (en) * 2018-01-29 2019-07-02 辽宁省蚕业科学研究所 A kind of Mongolian oak green branch cuttage breeding method
CN108094033B (en) * 2018-01-29 2019-07-02 辽宁省蚕业科学研究所 A kind of Quercus acutissima green branch cuttage breeding method
CN109618805A (en) * 2019-02-27 2019-04-16 湖南春光九汇现代中药有限公司 A kind of implantation methods of evodia rutaecarpa
CN115380822A (en) * 2022-08-03 2022-11-25 中国林业科学研究院高原林业研究所 High mountain tannin extract tissue culture medium and tissue culture method
CN115735774B (en) * 2022-12-10 2024-03-26 河北省洪崖山国有林场(河北雄安新区白洋淀上游规模化林场) Tissue culture method of Quercus mongolica

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
HU211394B (en) * 1990-01-11 1996-02-28 Chinoin Gyogyszer Es Vegyeszet New culture medium for increase of oak plantcallus
ES2180385A1 (en) * 2000-08-01 2003-02-01 Univ Madrid Politecnica Production of cork haploid plants and embryos consists of gametic embryogenesis, germination, and accelerated overall growth
CN1899028A (en) * 2006-07-24 2007-01-24 中国科学院昆明植物研究所 Tissue culture method for tri-ridged oak

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
HU211394B (en) * 1990-01-11 1996-02-28 Chinoin Gyogyszer Es Vegyeszet New culture medium for increase of oak plantcallus
ES2180385A1 (en) * 2000-08-01 2003-02-01 Univ Madrid Politecnica Production of cork haploid plants and embryos consists of gametic embryogenesis, germination, and accelerated overall growth
CN1899028A (en) * 2006-07-24 2007-01-24 中国科学院昆明植物研究所 Tissue culture method for tri-ridged oak

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
M.C.San-jose.E.,et al..Shoot apex explants for induction of somatic embryogenesis in mature Quercus robur L.trees.《Plant cell rep》.2010,第29卷全文.
Plant regeneration through somatic embryogenesis from tissues of mutant oak trees:true-to-type conformity of plantlets by RAPD analysis;S.Valladares,et al.;《Plant cell rep》;20061231;第25卷;全文 *
S.Valladares,et al..Plant regeneration through somatic embryogenesis from tissues of mutant oak trees:true-to-type conformity of plantlets by RAPD analysis.《Plant cell rep》.2006,第25卷全文.
Shoot apex explants for induction of somatic embryogenesis in mature Quercus robur L.trees;M.C.San-jose.E.,et al.;《Plant cell rep》;20101231;第29卷;全文 *
张存旭等.栓皮栎茎段离体培养的研究.《西北植物学报》.2004,第24卷(第7期),全文.
栓皮栎茎段离体培养的研究;张存旭等;《西北植物学报》;20041231;第24卷(第7期);全文 *

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