CN106258989A - A kind of Semen Arecae Ke's tissue cultured seedling root induction method - Google Patents

A kind of Semen Arecae Ke's tissue cultured seedling root induction method Download PDF

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Publication number
CN106258989A
CN106258989A CN201610900829.6A CN201610900829A CN106258989A CN 106258989 A CN106258989 A CN 106258989A CN 201610900829 A CN201610900829 A CN 201610900829A CN 106258989 A CN106258989 A CN 106258989A
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root
semen arecae
bud
tissue cultured
culture
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黄文卫
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Liuzhou Tinkling Of Pieces Of Jade Leads To Science And Technology Ltd Co
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Liuzhou Tinkling Of Pieces Of Jade Leads To Science And Technology Ltd Co
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001—Culture apparatus for tissue culture
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008—Methods for regeneration to complete plants

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  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention discloses a kind of Semen Arecae Ke's tissue cultured seedling root induction method, choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 35～40d, it is inoculated in after choosing pruning further in pre-root media, proceeds to again root media is cultivated after 30～35d pre-root culture.Present invention reduces Semen Arecae Ke's subculture blastogenesis root cycle, rooting rate is high, and root system is many, quality is good, and tissue cultured seedling transplanting survival rate of taking root is high, can realize Semen Arecae Ke and organize training industrialization nursery, construction for artificial clone woods provides high quality seedling, has preferable economic benefit, social benefit and ecological benefits.

Description

A kind of Semen Arecae Ke's tissue cultured seedling root induction method
Technical field
The present invention relates to Semen Arecae Ke's vegetative propagation technique, especially a kind of Semen Arecae Ke's tissue cultured seedling root induction method.
Background technology
Semen Arecae Ke (Lithocarpus areca (Hick. et A. Camus) A. Camus), is Fagaceae (Fagaceae) Lithocarpus (Lithocarpus) arbor, high 10～15 meters, sprig canescence, without hair, there is hole skin.Leaf papery, drapes over one's shoulders Aciculiform or narrow oblong, long 13～25 centimetres, wide 3.5～5.5 centimetres, the narrow point in two ends, base portion is downward, and minority is arranged at leaf margin top The shallow sharp mouth split or have full edge concurrently, middle arteries and veins in blade face microprotrusion, the every limit of lateral vein 9～15, in blade face micro-pits, offshoot is obvious, Two sides is homochromy, without having feathering on hair or arteries and veins axil；Petiole length 0.5～1.5 centimetres.Tassel locusta list fringe axil is raw, rare short branch Form coniform, long 5～8 centimetres, to spend many and intensive, filigree is very thin, the longest, and rachis is very thin；Female inflorescence length 4～10 centimetres, Often androgyny, female flower be born in rachis lower in, therefore infructescence is the shortest；Every 3～5 clusters of female flower, usual 1 bow structure is real, gynoecium Ovum shape triangle；Squamella short-term shape when acorn-cup children is tender, extends to 8 millimeters time ripe by long 2～4 millimeters；Nut is olive-shaped ellipse Circle, or conico-acuminate, high 40～50 millimeters, wide 20～35 millimeters, 3 longitudinal directions ridge rib the most in obtuse angle is arranged at top, and top is sharp, Chestnut, without hair, really wall thickness 2～3 millimeters, base portion is smooth, and areola caves in, deep 2～3 millimeters, bore 8～15 millimeters.Florescence 10 Month, really November next year is ripe.Semen Arecae Ke originates in municipal western part of Zhuang National Minority of China (Napo County, Longzhou), the southeast, Yunnan Province (Malipo), is grown on height above sea level 800 meters to 1, and the area of 500 meters is grown in evergreen broadleaf forest more, and North Vietnam also has point Cloth.
At present, Semen Arecae Ke's resource is still in wild distribution, but along with the demand of protection Plant Diversity, scale people Work cultivation Semen Arecae Ke's resource is imperative.Tissue culture technique is a kind of fast asexual propagation technology, selects Semen Arecae Ke You respectable family System or clone are material, breed nursery stock by method for tissue culture, both can meet the quantitative requirement in production, and can protect again Hold maternal character.But during group training, usually occur that rooting rate is low, disunity of taking root, root quantity are few and root system not Sturdy problem.
Summary of the invention
It is an object of the invention to provide a kind of rooting rate that can improve Semen Arecae Ke's tissue cultured seedling, root quantity and root system are sprouted Send out Semen Arecae Ke's tissue cultured seedling root induction method of regularity.
In order to realize foregoing invention purpose, technical scheme is as follows:
A kind of Semen Arecae Ke's tissue cultured seedling root induction method, chooses proliferation and subculture Semen Arecae Ke's seedling, first carries out pre-root culture, then Carrying out root culture again, be placed in control environment cultivation, it is thus achieved that band root tissue cultured seedling, main operational steps is as follows:
(1) draw materials: choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 35～40d, after sterilization bottle appearance, super In sterilized space on clean workbench, choose robust growth, highly 1～the simple bud of 2cm in subculture bud clump, at joint lower 2～3mm Shear, remove radical leaves and petiole, standby；
(2) pre-root culture: the simple bud after step (1) being pruned is inoculated in pre-root media, at specific photo-thermal reaction In carry out pre-root culture；
(3) root culture: until the simple bud in step (2) after 30～35d pre-root culture, simple bud is transferred into root culture Base, carries out root culture in specific photo-thermal reaction.
Above-described its material content of pre-root media is: 1/2 modified MS medium+NAA 1.0～2.0mg/L+ Vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/L.
Its material content of above-described root media is: 1/2 modified MS medium+IBA 1.0～2.0mg/L+ ABT1#0.5～2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium: KNO3915 mg/L；NH4NO3790 mg/L； CaCl2·2H2O 260 mg/L；MgSO4·7H2O 395 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 320 mg/ L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/ L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B1 1.0 mg/ L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Photo-thermal reaction described in above step (2) and step (3) is: temperature 20 ± 1 DEG C, humidity 40～45%, intensity of illumination 2000～2500lux, illumination 15～16h/d.
The present invention has the advantage that and has the beneficial effect that:
1, the present invention uses 1/2 modified MS medium as minimal medium, and emphasis have adjusted the phase taken root with Semen Arecae Ke simultaneously The trace element closed and organic principle so that culture medium is more scientific, more targetedly, is more easy to promote Semen Arecae Ke's subculture blastogenesis root, Effect is notable.
2, the present invention uses low concentration NAA and ascorbic acid (VC) that subculture simple bud carries out pre-training of taking root before root induction Supporting, carry out root culture the most again, tissue cultured seedling root of hair can be made unified, root of hair speed is fast, and root system is sturdy and quantity is more.
3, the present invention proceeds to root culture proliferation and subculture simple bud after 30～35d time pre-root culture, i.e. up to To pre-root culture effect, turn avoid seedling and grow root system in the pre-root culture stage and make later stage root culture root of hair the most whole Together.
4, the present invention under specific light and temperature condition, carry out pre-taking root and root culture, adapt to the life of Semen Arecae Ke's tissue cultured seedling Long characteristic, promotes the growth of nursery stock.
5, present invention reduces Semen Arecae Ke's subculture blastogenesis root cycle, rooting rate is high, and root system is many, and quality is good, tissue cultured seedling of taking root Transplanting survival rate is high, can realize Semen Arecae Ke and organize training industrialization nursery, and the construction for artificial clone woods provides high quality seedling, has Preferably economic benefit, social benefit and ecological benefits.
Detailed description of the invention
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1:
Choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 35～36d, after sterilization bottle appearance, at superclean bench On sterilized space in, choose robust growth, highly 1～the simple bud of 2cm in subculture bud clump, in the lower 2～3mm places shearing of joint, clearly Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 40%, Intensity of illumination 2000lux, carries out pre-root culture under conditions of illumination 16h/d.Wherein said its raw material of pre-root media Content is: 1/2 modified MS medium+NAA 1.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/ L。
Simple bud, after 30～35d pre-root culture, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet Degree 40%, intensity of illumination 2000lux, carry out root culture under conditions of illumination 16h/d.Its raw material of described root media contains Amount is: 1/2 modified MS medium+IBA 1.0mg/L+ABT1#0.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium: KNO3915 mg/L；NH4NO3790 mg/L； CaCl2·2H2O 260 mg/L；MgSO4·7H2O 395 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 320 mg/ L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/ L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B1 1.0 mg/ L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 2:
Choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 36～37d, after sterilization bottle appearance, at superclean bench On sterilized space in, choose robust growth, highly 1～the simple bud of 2cm in subculture bud clump, in the lower 2～3mm places shearing of joint, clearly Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 40%, Intensity of illumination 2500lux, carries out pre-root culture under conditions of illumination 15h/d.Wherein said its raw material of pre-root media Content is: 1/2 modified MS medium+NAA 1.5mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/ L。
Simple bud, after 30～35d pre-root culture, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet Degree 40%, intensity of illumination 2500lux, carry out root culture under conditions of illumination 15h/d.Its raw material of described root media contains Amount is: 1/2 modified MS medium+IBA 1.0mg/L+ABT1#1.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium: KNO3915 mg/L；NH4NO3790 mg/L； CaCl2·2H2O 260 mg/L；MgSO4·7H2O 395 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 320 mg/ L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/ L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B1 1.0 mg/ L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 3:
Choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 37～38d, after sterilization bottle appearance, at superclean bench On sterilized space in, choose robust growth, highly 1～the simple bud of 2cm in subculture bud clump, in the lower 2～3mm places shearing of joint, clearly Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 45%, Intensity of illumination 2000lux, carries out pre-root culture under conditions of illumination 16h/d.Wherein said its raw material of pre-root media Content is: 1/2 modified MS medium+NAA 2.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/ L。
Simple bud, after 30～35d pre-root culture, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet Degree 45%, intensity of illumination 2000lux, carry out root culture under conditions of illumination 16h/d.Its raw material of described root media contains Amount is: 1/2 modified MS medium+IBA 2.0mg/L+ABT1#1.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium: KNO3915 mg/L；NH4NO3790 mg/L； CaCl2·2H2O 260 mg/L；MgSO4·7H2O 395 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 320 mg/ L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/ L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B1 1.0 mg/ L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 4:
Choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 39～40d, after sterilization bottle appearance, at superclean bench On sterilized space in, choose robust growth, highly 1～the simple bud of 2cm in subculture bud clump, in the lower 2～3mm places shearing of joint, clearly Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 45%, Intensity of illumination 2500lux, carries out pre-root culture under conditions of illumination 15h/d.Wherein said its raw material of pre-root media Content is: 1/2 modified MS medium+NAA 2.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/ L。
Simple bud, after 30～35d pre-root culture, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet Degree 45%, intensity of illumination 2500lux, carry out root culture under conditions of illumination 15h/d.Its raw material of described root media contains Amount is: 1/2 modified MS medium+IBA 2.0mg/L+ABT1#2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium: KNO3915 mg/L；NH4NO3790 mg/L； CaCl2·2H2O 260 mg/L；MgSO4·7H2O 395 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 320 mg/ L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/ L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B1 1.0 mg/ L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.

Claims (5)

1. Semen Arecae Ke's tissue cultured seedling root induction method, it is characterised in that: choose proliferation and subculture Semen Arecae Ke's seedling, first carry out pre- Root culture, carries out root culture the most again, is placed in control environment cultivation, it is thus achieved that band root tissue cultured seedling, main operational steps is such as Under:
(1) draw materials: choose strong bud in routine group is trained and cultivate Semen Arecae Ke's subculture bud of 35～40d, after sterilization bottle appearance, super In sterilized space on clean workbench, choose robust growth, highly 1～the simple bud of 2cm in subculture bud clump, at joint lower 2～3mm Shear, remove radical leaves and petiole, standby；
(2) pre-root culture: the simple bud after step (1) being pruned is inoculated in pre-root media, at specific photo-thermal reaction In carry out pre-root culture；
(3) root culture: until the simple bud in step (2) after 30～35d pre-root culture, simple bud is transferred into root culture Base, carries out root culture in specific photo-thermal reaction.
A kind of Semen Arecae Ke's tissue cultured seedling root induction method the most according to claim 1, it is characterised in that: described pre-take root Its material content of culture medium is: 1/2 modified MS medium+NAA 1.0～2.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sugarcane Sugar 25g/L+ agar 5.0g/L.
A kind of Semen Arecae Ke's tissue cultured seedling root induction method the most according to claim 1, it is characterised in that: described training of taking root Supporting its material content of base is: 1/2 modified MS medium+IBA 1.0～2.0mg/L+ABT1#0.5～2.0 mg/L+ sucrose 25g/ L+ agar 5.0g/L.
4. according to the arbitrary a kind of described Semen Arecae Ke's tissue cultured seedling root induction method of Claims 2 or 3, it is characterised in that: described Basic composition is of modified MS medium: KNO3915 mg/L；NH4NO3790 mg/L；CaCl2·2H2O 260 mg/L； MgSO4·7H2O 395 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 mg/L；MnSO4·H2O 22.3 mg/ L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/L；Vitamin B60.5 mg/L； Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
A kind of Semen Arecae Ke's tissue cultured seedling root induction method the most according to claim 1, it is characterised in that: step (2) and step Suddenly the photo-thermal reaction described in (3) is: temperature 20 ± 1 DEG C, humidity 40～45%, intensity of illumination 2000～2500lux, illumination 15～ 16h/d。
CN201610900829.6A 2016-10-17 2016-10-17 A kind of Semen Arecae Ke's tissue cultured seedling root induction method Pending CN106258989A (en)

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Application publication date: 20170104