CN103782912B - 红桂木组织培养培养基 - Google Patents
红桂木组织培养培养基 Download PDFInfo
- Publication number
- CN103782912B CN103782912B CN201410070503.6A CN201410070503A CN103782912B CN 103782912 B CN103782912 B CN 103782912B CN 201410070503 A CN201410070503 A CN 201410070503A CN 103782912 B CN103782912 B CN 103782912B
- Authority
- CN
- China
- Prior art keywords
- component
- acid
- culture
- substratum
- cassia tree
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 241000365971 Cassia renigera Species 0.000 title claims abstract description 18
- 239000001963 growth medium Substances 0.000 title abstract description 13
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 20
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 20
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 20
- 239000012879 subculture medium Substances 0.000 claims description 14
- 239000005648 plant growth regulator Substances 0.000 claims description 12
- 239000012153 distilled water Substances 0.000 claims description 11
- 239000011573 trace mineral Substances 0.000 claims description 11
- 235000013619 trace mineral Nutrition 0.000 claims description 11
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 claims description 10
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 10
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 10
- 239000004471 Glycine Substances 0.000 claims description 10
- 229910019142 PO4 Inorganic materials 0.000 claims description 10
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 10
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 10
- 239000004327 boric acid Substances 0.000 claims description 10
- LLSDKQJKOVVTOJ-UHFFFAOYSA-L calcium chloride dihydrate Chemical compound O.O.[Cl-].[Cl-].[Ca+2] LLSDKQJKOVVTOJ-UHFFFAOYSA-L 0.000 claims description 10
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 10
- 229910000366 copper(II) sulfate Inorganic materials 0.000 claims description 10
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 claims description 10
- 229940064880 inositol 100 mg Drugs 0.000 claims description 10
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 10
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 10
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 10
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 10
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 10
- 229960003512 nicotinic acid Drugs 0.000 claims description 10
- 235000001968 nicotinic acid Nutrition 0.000 claims description 10
- 239000011664 nicotinic acid Substances 0.000 claims description 10
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 10
- 239000010452 phosphate Substances 0.000 claims description 10
- 239000011591 potassium Substances 0.000 claims description 10
- 229910052700 potassium Inorganic materials 0.000 claims description 10
- 235000007715 potassium iodide Nutrition 0.000 claims description 10
- 229960004839 potassium iodide Drugs 0.000 claims description 10
- 239000004323 potassium nitrate Substances 0.000 claims description 10
- 235000010333 potassium nitrate Nutrition 0.000 claims description 10
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 claims description 10
- 229960004172 pyridoxine hydrochloride Drugs 0.000 claims description 10
- 235000019171 pyridoxine hydrochloride Nutrition 0.000 claims description 10
- 239000011764 pyridoxine hydrochloride Substances 0.000 claims description 10
- RWVGQQGBQSJDQV-UHFFFAOYSA-M sodium;3-[[4-[(e)-[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]-2-methylcyclohexa-2,5-dien-1-ylidene]methyl]-n-ethyl-3-methylanilino]methyl]benzenesulfonate Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C(=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=2C(=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=C1 RWVGQQGBQSJDQV-UHFFFAOYSA-M 0.000 claims description 10
- 229940088594 vitamin Drugs 0.000 claims description 10
- 235000013343 vitamin Nutrition 0.000 claims description 10
- 239000011782 vitamin Substances 0.000 claims description 10
- 229930003231 vitamin Natural products 0.000 claims description 10
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 10
- RZLVQBNCHSJZPX-UHFFFAOYSA-L zinc sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Zn+2].[O-]S([O-])(=O)=O RZLVQBNCHSJZPX-UHFFFAOYSA-L 0.000 claims description 10
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 claims description 5
- 239000005972 6-Benzyladenine Substances 0.000 claims description 5
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 5
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 5
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000012452 mother liquor Substances 0.000 description 15
- 230000001954 sterilising effect Effects 0.000 description 9
- 235000013399 edible fruits Nutrition 0.000 description 6
- 238000004659 sterilization and disinfection Methods 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 239000003104 tissue culture media Substances 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 238000011031 large-scale manufacturing process Methods 0.000 description 3
- 241000218232 Artocarpus Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241001446187 Kermes Species 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- CXORMDKZEUMQHX-UHFFFAOYSA-N kermesic acid Chemical compound O=C1C2=C(O)C(O)=CC(O)=C2C(=O)C2=C1C=C(O)C(C(O)=O)=C2C CXORMDKZEUMQHX-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000008774 maternal effect Effects 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 241001471144 Artocarpus nitidus subsp. lingnanensis Species 0.000 description 1
- 235000021537 Beetroot Nutrition 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241001411320 Eriogonum inflatum Species 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229960002523 mercuric chloride Drugs 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- -1 organism Substances 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 235000019614 sour taste Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Landscapes
- Hydroponics (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明公开了一种红桂木组织培养培养基,该培养基包括继代培养基和生根培养基。首次采用组织培养的方式进行种苗无性繁殖,该技术比通过种子繁殖更快,可达到规模化生产的要求,而且能有效保持种苗的优良遗传性状,保证了种苗的品质和质量。
Description
技术领域
本发明涉及组织培养基,尤其涉及一种红桂木组织培养培养基。
背景技术
红桂木(Artocarpus nitidus ssp.Lingnanensis(Merr.)Jarr.)又名狗果树、大叶胭脂、胭脂木等,属桑科桂木属常绿乔木,喜高温多湿气候,较耐旱,主要分布于广东中部至西南部、广西、海南、福建南部。其种子富含红桂木凝集素,可用于疾病检测治疗,临床应用潜力很大;果酸甜可口,可生津止渴,开胃化痰,消食敛气,药用价值颇高。木材坚硬,纹理通直,可供建筑、家具及器具等用。红桂木树形优美,枝叶繁茂,树干通直,抗污染及抗火性较强,适应性亦强,绿化效果好,苗木供不应求,特别是中大径级苗木更为匮乏,是一种应用前景广阔的优良乡土树种。
红桂木常用种子播种育苗,繁殖较慢,满足不了市场的需求,通过组织培养育苗则能提高繁殖速度,保证苗木质量,达到规模化生产的要求。有关红桂木树的组培育苗技术方面的研究目前还未见有报道。
发明内容
本发明的目的是克服现有技术的不足,提供一种红桂木组织培养培养基,该培养基包括继代培养基和生根培养基,其中:
每升(L)继代培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾1260mg/L、硝酸铵1320mg/L、二水氯化钙440mg/L、七水硫酸镁370mg/L、磷酸二氢钾170mg/L、七水硫酸亚铁27.8mg/L、乙二胺四乙酸二钠37.3mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/L、半胱氨酸15mg/L;
(4)植物生长调节剂:6-苄基腺嘌呤0.3-1.0mg/L、萘乙酸0.1-0.2mg/L;
余量为蒸馏水。
每升(L)生根培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾475mg/L、硝酸铵412.5mg/L、二水氯化钙110mg/L、七水硫酸镁92.5mg/L、磷酸二氢钾42.5mg/L、乙二胺四乙酸二钠37.3mg/L、七水硫酸亚铁27.8mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/;
(4)植物生长调节剂:ABT6号0.4-0.6mg/L、吲哚乙酸0.1-0.3mg/L;
余量为蒸馏水。
本发明红桂木组织培养基中的继代、生根培养基的配制方法为:
1母液的配制与保存
1.1为便于取样,宜先配制各种母液,分为大量元素母液、微量元素母液、有机物母液和各种植物生长调节剂母液。蔗糖、琼脂不宜配成母液,需要时直接称样;
1.2大量元素母液浓度成100倍溶液,有机物、微量元素母液配成200倍溶液,植物生长调节试剂母液的浓度配成1mg/ml;
1.3母液选用无菌的蒸馏水、去离子水或超纯水配制,大量生产时用煮沸过的水;
1.4配制大量元素母液时,每个组分应单独溶解,后按氮、钙、镁、磷的顺序逐个混合,否则容易引起沉淀;
1.5微量元素、有机物、植物生长调节剂母液应用棕色瓶装好并置于冰箱中保存;
1.6母液配制后应及时使用,贮存时间不宜超过1个月;
1.7发现母液有沉淀,或有微生物生长,或有藻类生长,应废弃不用。
2培养基的配制
2.1依照培养基配方,按比例量取各种母液;
2.2在定量容器内放入准备配制培养基总量的约1/2以上的纯净水,并加入适量的糖,然后边搅拌边逐一加入所需量的母液;
2.3琼脂可加热熔化后加入,如有搅拌设备时也可直接加入琼脂粉,然后用纯净水补足所需配制培养基的总量,搅拌均匀即可;
2.4培养基中的糖,在大量组培苗生产中,一般可用市售白糖,但最好是用蔗糖而不要用甜菜糖;
2.5用1.0摩尔/升的盐酸或1.0摩尔/升的氢氧化钠调节培养基的pH值至5.8;
2.6配制好的培养基要尽快分装到培养容器中,以免培养基凝固或变稠而难以分装;
2.7分装培养基时应注意避免培养基粘在瓶口上,如果粘有培养基,在盖瓶盖或瓶塞前必须用干净纱布擦净瓶口。
3培养基消毒灭菌
3.1将分装好的培养基装于高压灭菌锅内消毒;
3.2加热初期,当消毒锅消毒室的气压达0.05Mpa时,打开冷凝阀,排尽消毒室内冷空气;
3.3当消毒室内气压达到0.11Mpa、温度达121℃时,开始计时,保持温度与压力消毒15-20分钟;
3.4关闭加热电源开关,按慢排气方式排放热气,待消毒室内的气压降至大气压时打开消毒锅盖或门,取出培养基;
3.5培养基应置于空气少流动和少灰尘的干净环境冷却,否则在降温进气过程中导致霉菌孢子进入培养器皿,产生霉菌污染。
4培养基储藏
4.1培养基应尽量现配现用;
4.2可在空气干净且不流动的环境短时间储藏培养基,但储藏超过1个月的培养基应废弃不用。
本发明红桂木组织培养的方法为:
1.无菌芽的获取
胭脂树果实为肉质果,7-8月份成熟,当果实由青色转为黄色带红,果肉变软时采摘,将果实揉搓,用清水洗净后得种子,并将其及时置于阴凉处晾干,剥去外壳。在超净台上用75%酒精浸泡消毒10s,无菌水漂洗2-3次,再用0.1%升汞溶液消毒15min,无菌水漂洗5次。将经消毒处理的种子接种在诱导培养基上进行初始培养,诱导种子发芽以获取无菌材料。外植体初次培养时,需全暗培养10天左右,再放到弱光下进行培养,大约经过20天左右的种子开始萌芽,然后再移至见光的地方培养。
2.芽的增殖培养
将诱导获得的无菌芽接种至继代培养基上进行芽的继代增殖,一般40天继代一次,继代材料均放在培养室内培养,培养室温度为(25±2)℃,光照12h.d-1,光照度2000lx。
3.生根的诱导
芽条长至约2-3cm时,将较健壮的芽苗由丛生芽上单个切下转至生根培养基上诱导生根,其余芽苗转至继代培养基上继续进行增殖培养。生根培养需要在弱光下培养15天左右,此时小苗的基部形成白色突起,并逐渐伸长,20天后可形成明显根系,逐渐长成完整的小植株,此时期宜逐步增加光照使小苗变得粗壮。
4.生根苗的移栽
待生根小苗充分木质化,叶片舒展,叶色浓绿,茎轴伸长,苗高约3-4cm时即可进行大棚移栽。移栽时先取出小苗,洗净基部的培养基后,移栽于经0.1%高锰酸钾消毒过用营养袋装好的育苗基质上,移栽后浇透定根水,保持一定的温度和湿度,并要定期进行喷药进行防病处理。
本发明的优点是:
1、首次采用组织培养的方法对红桂木进行苗木繁殖,该技术比通过种子繁殖更快,可达到规模化生产的要求,而且能有效保持母本的优良遗传性状,保证了种苗的品质和质量;
2、以继代培养基进行红桂木增殖培养,培养温度保持在25-28℃左右,继代苗年繁殖系数为3.512,继代苗分化芽多,芽健壮,生长整齐,叶色浓绿;
3、用生根培养基进行生根苗培养生根率达92.0%,温度为25-28℃时,培养20天左右可出根,根系发达,平均出根量有3-5条,40天可移栽,移栽成活率达91.0%。
具体实施方式
下面以实施例对本发明作进一步说明,但本发明并不局限于这些实施例。
实施例1:
红桂木组织培养的培养基配方,该培养基包括继代培养基和生根培养基,其中:
每升(L)继代培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾1260mg/L、硝酸铵1320mg/L、二水氯化钙440mg/L、七水硫酸镁370mg/L、磷酸二氢钾170mg/L、七水硫酸亚铁27.8mg/L、乙二胺四乙酸二钠37.3mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/L、半胱氨酸15mg/L;
(4)植物生长调节剂:6-苄基腺嘌呤1.0mg/L、萘乙酸0.2mg/L;
余量为蒸馏水。
(1)大量元素:硝酸钾475mg/L、硝酸铵412.5mg/L、二水氯化钙110mg/L、七水硫酸镁92.5mg/L、磷酸二氢钾41mg/L、乙二胺四乙酸二钠37.3mg/L、七水硫酸亚铁27.8mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/;
(4)植物生长调节剂:ABT6号0.5mg/L、吲哚乙酸0.3mg/L;
余量为蒸馏水。
实施例2:
红桂木组织培养的培养基配方,该培养基包括继代培养基和生根培养基,其中:
每升(L)继代培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾1260mg/L、硝酸铵1320mg/L、二水氯化钙440mg/L、七水硫酸镁370mg/L、磷酸二氢钾170mg/L、七水硫酸亚铁27.8mg/L、乙二胺四乙酸二钠37.3mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/L、半胱氨酸15mg/L;
(4)植物生长调节剂:6-苄基腺嘌呤0.5mg/L、萘乙酸0.15mg/L;
余量为蒸馏水。
(1)大量元素:硝酸钾475mg/L、硝酸铵412.5mg/L、二水氯化钙110mg/L、七水硫酸镁92.5mg/L、磷酸二氢钾41mg/L、乙二胺四乙酸二钠37.3mg/L、七水硫酸亚铁27.8mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/;
(4)植物生长调节剂:ABT6号0.4mg/L、吲哚乙酸0.2mg/L;
余量为蒸馏水。
实施例3:
红桂木组织培养的培养基配方,该培养基包括继代培养基和生根培养基,其中:
每升(L)继代培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾1260mg/L、硝酸铵1320mg/L、二水氯化钙440mg/L、七水硫酸镁370mg/L、磷酸二氢钾170mg/L、七水硫酸亚铁27.8mg/L、乙二胺四乙酸二钠37.3mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/L、半胱氨酸15mg/L;
(4)植物生长调节剂:6-苄基腺嘌呤0.3mg/L、萘乙酸0.1mg/L;
余量为蒸馏水。
(1)大量元素:硝酸钾475mg/L、硝酸铵412.5mg/L、二水氯化钙110mg/L、七水硫酸镁92.5mg/L、磷酸二氢钾41mg/L、乙二胺四乙酸二钠37.3mg/L、七水硫酸亚铁27.8mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/;
(4)植物生长调节剂:ABT6号0.6mg/L、吲哚乙酸0.1mg/L;
余量为蒸馏水。
通过以上实施例对红桂木进行组培繁殖,用继代培养基进行增殖培养,培养温度保持在25-28℃左右,继代苗年繁殖系数为3.512,继代苗分化芽多,芽健壮,生长整齐,叶色浓绿;用生根培养基进行生根苗培养生根率达92.0%,温度为25-28℃时,培养20天左右可出根,根系发达,平均出根量有3-5条,40天可移栽,移栽成活率达91.0%。采用本发明的技术比通过种子繁殖更快,可达到规模化生产的要求,而且能有效保持母本的优良遗传性状,保证了种苗的品质和质量。
Claims (1)
1.一种红桂木组织培养的培养基,该培养基包括继代培养基和生根培养基,其特征在于:
每升(L)继代培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾1260mg/L、硝酸铵1320mg/L、二水氯化钙440mg/L、七水硫酸镁370mg/L、磷酸二氢钾170mg/L、七水硫酸亚铁27.8mg/L、乙二胺四乙酸二钠37.3mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/L、半胱氨酸15mg/L;
(4)植物生长调节剂:6-苄基腺嘌呤0.3-1.0mg/L、萘乙酸0.1-0.2mg/L;
余量为蒸馏水;
每升(L)生根培养基的组分及各组分的含量如下:
(1)大量元素:硝酸钾475mg/L、硝酸铵412.5mg/L、二水氯化钙110mg/L、七水硫酸镁92.5mg/L、磷酸二氢钾41mg/L、乙二胺四乙酸二钠37.3mg/L、七水硫酸亚铁27.8mg/L;
(2)微量元素:四水硫酸锰22.3mg/L、七水硫酸锌8.6mg/L、硼酸6.2mg/L、二水钼酸钠0.25mg/L、碘化钾0.83mg/L、五水硫酸铜0.025mg/L、六水氯化钴0.025mg/L;
(3)有机物:肌醇100mg/L、甘氨酸2.0mg/L、盐酸吡哆醇(VB6)0.5mg/L、烟酸0.5mg/L、盐酸硫胺素(VB1)0.1mg/;
(4)植物生长调节剂:ABT6号0.4-0.6mg/L、吲哚乙酸0.1-0.3mg/L;
余量为蒸馏水。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410070503.6A CN103782912B (zh) | 2014-02-28 | 2014-02-28 | 红桂木组织培养培养基 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410070503.6A CN103782912B (zh) | 2014-02-28 | 2014-02-28 | 红桂木组织培养培养基 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103782912A CN103782912A (zh) | 2014-05-14 |
CN103782912B true CN103782912B (zh) | 2015-04-22 |
Family
ID=50659300
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410070503.6A Expired - Fee Related CN103782912B (zh) | 2014-02-28 | 2014-02-28 | 红桂木组织培养培养基 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN103782912B (zh) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104663464A (zh) * | 2015-03-31 | 2015-06-03 | 桂林得坤生物科技股份有限公司 | 柿树组织培养培养基 |
CN106900555B (zh) * | 2017-03-21 | 2018-10-09 | 钦州市林业科学研究所 | 阳春砂试管分株培养基及一次成苗组培分株快繁方法 |
CN111072422A (zh) * | 2020-01-16 | 2020-04-28 | 河南华薯农业科技有限公司 | 一种普薯32的专用培养基及其制备方法 |
CN115093276A (zh) * | 2022-05-13 | 2022-09-23 | 和田博正农业科技有限公司 | 一种广谱生根剂的配制及使用方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103125383A (zh) * | 2011-12-02 | 2013-06-05 | 云南省热带作物科学研究所 | 一种建立橡胶树未授粉胚珠再生体系的方法 |
-
2014
- 2014-02-28 CN CN201410070503.6A patent/CN103782912B/zh not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103125383A (zh) * | 2011-12-02 | 2013-06-05 | 云南省热带作物科学研究所 | 一种建立橡胶树未授粉胚珠再生体系的方法 |
Non-Patent Citations (2)
Title |
---|
不同基质和生长调节剂对红桂木光合与水分利用效率的影响;陈香等;《亚热带植物科学》;20111231;第40卷(第1期);第5-8页 * |
牛大力组织培养瓶内复壮瓶外生根快繁技术;时群等;《林业实用技术》;20091231(第12期);第30页 * |
Also Published As
Publication number | Publication date |
---|---|
CN103782912A (zh) | 2014-05-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104145816B (zh) | 白芨组培育苗方法 | |
CN102577976B (zh) | 一种构树简便组培方法 | |
CN103651122B (zh) | 一种白及原球茎诱导培养基 | |
CN102823503B (zh) | 红掌以芽繁芽组织培养培养基 | |
CN102884981B (zh) | 两面针组织培养培养基 | |
CN103798144A (zh) | 红椿组织培养培养基 | |
CN105010109A (zh) | 罗汉果的水培方法 | |
CN103798145A (zh) | 扁桃斑鸠菊组织培养培养基 | |
CN103782912B (zh) | 红桂木组织培养培养基 | |
CN105237090A (zh) | 桑黄固态菌种转液态培养基的培育方法 | |
CN101273709A (zh) | 一种铁皮石斛快繁的组培方法 | |
CN106900555B (zh) | 阳春砂试管分株培养基及一次成苗组培分株快繁方法 | |
CN102150619A (zh) | 辣木胚愈伤组织诱导和植株再生方法 | |
CN104303840B (zh) | 一种盘装金针菇的栽培方法 | |
CN103931496A (zh) | 一种曲茎石斛组培快繁培养基及组培快繁方法 | |
CN104770293A (zh) | 白芨开放式组培育苗方法 | |
CN105309301A (zh) | 鼓槌石斛种苗商品化生产方法 | |
CN103598093B (zh) | 一种蓝莓胚状体的诱导方法 | |
CN109874676A (zh) | 一种百香果脱毒苗的快速繁育方法 | |
CN103392594A (zh) | 金心也门铁组培苗生根诱导技术 | |
CN110214692A (zh) | 一种植物非固体组培培养基及植物非固体组培方法 | |
CN105746358A (zh) | 栓翅卫矛组培配方和培养方法 | |
CN109328985A (zh) | 一种促进霍山石斛种子萌发的培养基及播种方法 | |
CN103858764B (zh) | 鹅不食草组培快繁培养基及一次成苗组培快繁方法 | |
CN104351057A (zh) | 无菌即食生菜的种植方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150422 |
|
CF01 | Termination of patent right due to non-payment of annual fee |