CN102577952A - Tissue culturing method for quercus virginiana - Google Patents

Tissue culturing method for quercus virginiana Download PDF

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CN102577952A
CN102577952A CN2012100297273A CN201210029727A CN102577952A CN 102577952 A CN102577952 A CN 102577952A CN 2012100297273 A CN2012100297273 A CN 2012100297273A CN 201210029727 A CN201210029727 A CN 201210029727A CN 102577952 A CN102577952 A CN 102577952A
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culture
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explant
seedling
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CN102577952B (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 Virginia oak method for tissue culture
Technical field
The invention belongs to method for plant tissue culture, be specially a kind of Virginia oak method for tissue culture.
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, proterties such as leaf color, shape, size, quality have very strong plasticity with influences such as developmental stage, environment, the big 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, promptly falls after ripe, and seed vitality is lower.Virginia oak is liked well-drained sandy soil, can salt spray resistance and soil with high salinity, and responsive to soil moisture, can stand moderate arid, clay also can be grown, and can tolerate minimum-13 ℃ low temperature.Virginia oak juvenile growth rapid speed, seedling can grow to 4 feet (1.2m) in 1 year, and along with growing up of nursery stock, growth rate can reduce.Adult not oak in the growth of hard-core space on average can reach the 15m height, and the diameter of a cross-section of a tree trunk 1.3 meters above the ground can reach 91-122cm, and the tree crown scope can reach more than the 46m, and the life-span reaches hundreds of years.Virginia oak mainly is distributed in U.S.'s southeastern coast and Gulf of Mexico seashore, i.e. downstate, the Georgia State, Florida State, Louisiana State from Virginia are until the middle and south, Texas one line; Breathing out the Ma Zhou west and south at the Russia carat also has fragmentary distribution with Mexico's Northeast Mountain Areas, is the important composition seeds in coastal hard wealthy woods of the U.S. and shrubbery forest land, is widely used as the city in U.S.'s southeastern coast and shades and set and ornamental trees and shrubs.
The Virginia oak seed that Inst. of Subtropical Forestry, Chinese Academy of Forestry Sciences introduces from Missouri, USA, the plant trunk of Cheng Miaohou about 5% is obvious, and high growth is outstanding, and branch is sparse, and is crown narrow and small; Most nursery stocks then 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 Virginia oak is introduced a fine variety; All can grow in subtropics, torrid areas such as China Zhejiang, Shanghai, Jiangsu, Fujian, have very strong salt tolerant alkali ability owing to Virginia oak simultaneously, in addition well developed root system; Germinating property is strong; Branch tool toughness has the ability of resisting hurricane, therefore is widely used in the coast protection forest construction in China Yangtze River Delta area; Having remedied 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 the present southeastern coast protection forest system.Simultaneously, acorn has attracted large quantities of birds and animal to compile, and has enriched the bio-diversity of coast protection forest system.Virginia oak also can be used for public leisure place greenings such as garden, park, farm, and its tree crown is wide, and the leaf look vivid, is the habitat of good shade, lie fallow place and animal of people.
Virginia oak is annual in the original producton location can to produce a large amount of rubber fruits, therefore adopts seminal propagation mostly, and the clone of having only minority nursery stock company to rely on vegetative propagation to obtain various trait is 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, as long as temperature is suitable can be sprouted, therefore in import kind subprocess; Owing to reasons such as mechanical damages; Make greatly seed after sprouting, can not get sowing in time and devitalization causes emergence rate low, seedling cost is higher.Inst. of Subtropical Forestry, Chinese Academy of Forestry Sciences introduces Virginia oak at home first and carries out relevant research; And carried out Virginia oak vegetative propagation technique research; Aspect cuttage propagation method of Virginia oak, obtained very quantum jump (ZL200910097684.0), survival rate can be up to 80-85%.
Summary of the invention
To the problems referred to above that exist in the prior art; The objective of the invention is to design provides a kind of technical scheme that significantly improves the Virginia oak method for tissue culture of its reproduction coefficient; And shortened the Virginia oak growing-seedling period greatly; Solve the mode that the dependence on import seed is grown seedlings, practiced thrift cost.
Described a kind of Virginia oak method for tissue culture is characterized in that may further comprise the steps:
1) explant selection: 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, after 10-14 hour, with the alcohol disinfecting 25-35 second of 70-75%, mercuric chloride soaked 3-5 minute explant then again through the running water flushing, with aseptic water washing 2-3 time, placed aseptic vial subsequent use then;
3) initial culture: the stem with bud that the explant of will sterilizing is cut into 0.8-1.2cm inserts in the medium to be cultivated 25-35 days, gave illumination about 2500-3500 Lx, photoperiod 14-18hr/6-10hr, 25 ℃ ± 2 ℃ of temperature; Described initial culture base is made up 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 the initial culture base is inserted in the proliferated culture medium carry out enrichment culture, the same step 3) of condition of culture, cultivation cycle are 40-50 days; Described proliferated culture medium is made up of WPM or 1/4MS+6-benzyladenine 0.1-0.3mg/L+ growth regulatory substance methyl 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, inserts in WPM or the 1/4MS medium and cultivates 25-35 days, the same step 3) of condition of culture;
6) culture of rootage: bud healthy and strong after the strong seedling culture is cut the base portion brown material insert in the root media with callus and carry out culture of rootage, dark earlier to cultivate 1.5-2.5 all, and illumination cultivation 3.5-4.5 was taken root after week again, the same step 3) of condition of culture; Described root media is made up of WPM or 1/4MS+ growth regulatory substance 6-benzyladenine 0.8-1.2 mg/L+growth regulatory substance methyl 0.4-0.6 mg/L;
7) refining seedling 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 under the room temperature; Flush away root medium is transplanted and is equipped with on the seedbed of river sand matrix, keeps 20-25 ℃ of temperature; Humidity 85%-95%, suitably shading gets final product.
Described a kind of Virginia oak method for tissue culture is characterized in that in the 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, the explant that gather in the land for growing field crops, and other adds 1-2 and drips the Tween 80 sterilization;
Indoor cultivation: choose the Virginia oak seed of no small holes caused by worms, full grains, in running water, soaked 22-26 hour, discard floating seed; Soak with saturated bleaching powder supernatant then and carried out surface sterilization in 25-35 minute; Planting seed after the sterilization covers river sand in the container in the plastic containers of 30cm * 20cm * 8cm, river sand thickness is 4-6cm; Place illumination box to cultivate plastic containers; 25 ± 1 ℃ of temperature, intensity of illumination 11000-13000Lx, light/dark cycle is 14-18h/6-10h.
Described a kind of Virginia oak method for tissue culture is characterized in that adding sucrose 15-25g.L-1 respectively in described initial culture base, adventitious bud proliferation medium, strong seedling culture base, the root media, and agar 0.5-1.0% regulates PH to 5.5-5.9.
Described a kind of Virginia oak method for tissue culture; It is characterized in that need taking explant anti-browning measure in the described initial culture process of step 3), be specially: in the initial culture base, add the active carbon that adds 0.2-0.4% or 0.4-0.6%, or repeatedly switching repeatedly; Be after explant is inoculated for the first time; Cultivated 2-3 days, and be transferred to same medium, change 2-3 time like this.
Described a kind of Virginia oak method for tissue culture is characterized in that described WPM medium is by being made up 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 Virginia oak method for tissue culture is characterized in that described MS medium is made up 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) KI 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 Virginia oak method for tissue culture is characterized in that used 1/4MS medium is that macroelement is 1/4 of a MS minimal medium, and all the other composition trace elements, molysite, organic principle are with the MS minimal medium.
Above-mentioned a kind of Virginia oak method for tissue culture can keep maternal resistance and sight preferably, effectively improves the Virginia oak reproduction coefficient.Compare with seminal propagation, tissue culture reduces the dependence to 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 practice thrift cost in 4 months.
Embodiment
Below in conjunction with specific embodiment the present invention is described further.
Embodiment 1: a kind of Virginia oak method for tissue culture, and its incubation step is following:
(1) explant selection: explant picks up from the plant that the degeneration-resistant physiological ecological of Chinese forest-science academy inferior woods institute is tested indoor cultivation.
(2) explant sterilization: get stem with bud as explant; Explant after running water flushing 12 hours, again with the alcohol disinfecting of 70-75% 30 seconds (if the explant that gather in the land for growing field crops then adds 1-2 and drips Tween 80; To strengthen the infiltration of disinfectant); Mercuric chloride soaked 3-5 minute then, with aseptic water washing for several times, placed aseptic vial subsequent use then.
(3) initial culture: the explant of will sterilizing is cut into stem with bud about 1cm and inserts in WPM or the 1/4MS+ growth regulatory substance 6-benzyladenine 1.0mg/L medium and cultivated 30 days, gives illumination about 3000lx, photoperiod 16hr/8hr, 25 ℃ ± 2 ℃ of temperature.Because the Virginia oak explant contains than polyphenols, therefore in the initial culture process, need take the anti-browning measure, concrete measure comprises: the active carbon that in medium, adds 0.2-0.4% or 0.4-0.6%; Or repeatedly switching repeatedly; Be after explant is inoculated for the first time, cultivated 3 days, be transferred to same medium; Change 3 times like this, can effectively prevent brownization generation.
(4) adventitious bud proliferation is cultivated: the stem with bud that the growth sign is arranged in the initial culture base is inserted in WPM or the 1/4MS+ growth regulatory substance 6-benzyladenine 0.2mg/L+ growth regulatory substance methyl 0.5 mg/L medium carry out enrichment culture; Condition of culture is with the step 3) initial culture, 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, inserts in WPM or the 1/4MS medium and cultivates 30 days, and condition of culture is with the step 3) initial culture.
(6) culture of rootage: bud healthy and strong after the strong seedling culture cut in base portion brown material and callus access WPM or the 1/4MS+ growth regulatory substance 6-benzyladenine 1.0 mg/L+ growth regulatory substance methyls 0.5 mg/L medium carry out culture of rootage; Dark earlier the cultivation for 2 weeks; Illumination cultivation was taken root after 4 weeks again, and condition of culture is with the step 3) initial culture.
(7) refining seedling 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 3 days under the room temperature; Flush away 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 made up 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 made up 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) KI 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) explant is cut into the stem with bud of 0.8cm or 1.2cm in; Cultivated 25 days or 35 days; Give about 2500 Lx of illumination, 2600 Lx or 3500 Lx, photoperiod 14hr/6hr or 18hr/10hr, 6-benzyladenine 0.8mg/L or 1.2mg/L in the described initial culture base; Cultivation cycle is 40 days or 50 days in the step 4); 6-benzyladenine 0.1 or 0.3mg/L in the described proliferated culture medium, growth regulatory substance methyl 0.4 or 0.6 mg/L.Cultivation cycle is 25 or 35 days in the step 5); Dark earlier the cultivation for 1.5 week or 2.5 weeks in the step 6), again 3.5 week or 4.5 weeks of illumination cultivation, the growth regulatory substance 6-benzyladenine is 0.8 or 1.2 mg/L in the described root media, the growth regulatory substance methyl is 0.4 or 0.6 mg/L.Other step also can reach beneficial effect of the present invention with embodiment 1.

Claims (7)

1. Virginia oak method for tissue culture is characterized in that may further comprise the steps:
1) explant selection: 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, after 10-14 hour, with the alcohol disinfecting 25-35 second of 70-75%, mercuric chloride soaked 3-5 minute explant then again through the running water flushing, with aseptic water washing 2-3 time, placed aseptic vial subsequent use then;
3) initial culture: the stem with bud that the explant of will sterilizing is cut into 0.8-1.2cm inserts in the medium to be cultivated 25-35 days, gave illumination about 2500-3500 Lx, photoperiod 14-18hr/6-10hr, 25 ℃ ± 2 ℃ of temperature; Described initial culture base is made up 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 the initial culture base is inserted in the proliferated culture medium carry out enrichment culture, the same step 3) of condition of culture, cultivation cycle are 40-50 days; Described proliferated culture medium is made up of WPM or 1/4MS+6-benzyladenine 0.1-0.3mg/L+ growth regulatory substance methyl 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, inserts in WPM or the 1/4MS medium and cultivates 25-35 days, the same step 3) of condition of culture;
6) culture of rootage: bud healthy and strong after the strong seedling culture is cut the base portion brown material insert in the root media with callus and carry out culture of rootage, dark earlier to cultivate 1.5-2.5 all, and illumination cultivation 3.5-4.5 was taken root after week again, the same step 3) of condition of culture; Described root media is made up of WPM or 1/4MS+ growth regulatory substance 6-benzyladenine 0.8-1.2 mg/L+growth regulatory substance methyl 0.4-0.6 mg/L;
7) refining seedling 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 under the room temperature; Flush away root medium is transplanted and is equipped with on the seedbed of river sand matrix, keeps 20-25 ℃ of temperature; Humidity 85%-95%, suitably shading gets final product.
2. a kind of Virginia oak method for tissue culture as claimed in claim 1 is characterized in that in the 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, the explant that gather in the land for growing field crops, and other adds 1-2 and drips the Tween 80 sterilization;
Indoor cultivation: choose the Virginia oak seed of no small holes caused by worms, full grains, in running water, soaked 22-26 hour, discard floating seed; Soak with saturated bleaching powder supernatant then and carried out surface sterilization in 25-35 minute; Planting seed after the sterilization covers river sand in the container in the plastic containers of 30cm * 20cm * 8cm, river sand thickness is 4-6cm; Place illumination box to cultivate plastic containers; 25 ± 1 ℃ of temperature, intensity of illumination 11000-13000Lx, light/dark cycle is 14-18h/6-10h.
3. a kind of Virginia oak method for tissue culture as claimed in claim 1; It is characterized in that adding sucrose 15-25g.L-1 respectively in described initial culture base, adventitious bud proliferation medium, strong seedling culture base, the root media; Agar 0.5-1.0% regulates PH to 5.5-5.9.
4. a kind of Virginia oak method for tissue culture as claimed in claim 1; It is characterized in that need taking explant anti-browning measure in the described initial culture process of step 3), be specially: in the initial culture base, add the active carbon that adds 0.2-0.4% or 0.4-0.6%, or repeatedly switching repeatedly; Be after explant is inoculated for the first time; Cultivated 2-3 days, and be transferred to same medium, change 2-3 time like this.
5. a kind of Virginia oak method for tissue culture as claimed in claim 1 is characterized in that described WPM medium is by being made up 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
6. a kind of Virginia oak method for tissue culture as claimed in claim 1 is characterized in that described MS medium is made up 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) KI 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
7. a kind of Virginia oak method for tissue culture as claimed in claim 1 is characterized in that used 1/4MS medium is that macroelement is 1/4 of a 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 (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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CN103636506A (en) * 2013-12-24 2014-03-19 黑龙江省林业科学研究所 Shepherdia argentea caulicle regenerated plant induction culture medium and method for performing plant culture by utilizing culture medium
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
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
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
CN107047297A (en) * 2017-02-14 2017-08-18 唐春艳 A kind of rice sweet oak tissue-cultured seedling rooting induction method
CN108094033A (en) * 2018-01-29 2018-06-01 辽宁省蚕业科学研究所 A kind of Quercus acutissima green branch cuttage breeding method
CN108243817A (en) * 2018-01-29 2018-07-06 辽宁省蚕业科学研究所 A kind of Mongolian oak 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
CN115735774A (en) * 2022-12-10 2023-03-07 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

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Cited By (17)

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Publication number Priority date Publication date Assignee Title
CN103535280A (en) * 2013-10-29 2014-01-29 江苏省林业科学研究院 Water oak tissue culture breeding method
CN103548679A (en) * 2013-10-29 2014-02-05 江苏省林业科学研究院 Method for quercus nuttallii somatic embryogenesis
CN103548679B (en) * 2013-10-29 2015-02-18 江苏省林业科学研究院 Method for quercus nuttallii somatic embryogenesis
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
CN103636506A (en) * 2013-12-24 2014-03-19 黑龙江省林业科学研究所 Shepherdia argentea caulicle regenerated plant induction culture medium and method for performing plant culture by utilizing culture medium
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
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
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
CN107047297A (en) * 2017-02-14 2017-08-18 唐春艳 A kind of rice sweet oak tissue-cultured seedling rooting induction method
CN108094033A (en) * 2018-01-29 2018-06-01 辽宁省蚕业科学研究所 A kind of Quercus acutissima green branch cuttage breeding method
CN108243817A (en) * 2018-01-29 2018-07-06 辽宁省蚕业科学研究所 A kind of Mongolian oak green branch cuttage breeding 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
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
CN115735774A (en) * 2022-12-10 2023-03-07 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
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

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