CN107667868A - A kind of method of canna cultured in vitro - Google Patents

A kind of method of canna cultured in vitro Download PDF

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Publication number
CN107667868A
CN107667868A CN201711201037.0A CN201711201037A CN107667868A CN 107667868 A CN107667868 A CN 107667868A CN 201711201037 A CN201711201037 A CN 201711201037A CN 107667868 A CN107667868 A CN 107667868A
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canna
culture
bud
explant
culture medium
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李海飞
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Guangxi Beiliu Sen Porcelain Industry Co Ltd
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Guangxi Beiliu Sen Porcelain Industry Co Ltd
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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
    • A01H4/001—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)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention discloses a kind of method of canna cultured in vitro, including explant processing, Fiber differentiation, Multiplying culture and culture of rootage, rhizome of the canna underground with bud is chosen as explant, rhizome base portion with bud is cut into stem with bud, explant is carried out after cleaning sterilization processing, it is inoculated in culture in inducing culture and obtains the initial bud of canna, the initial bud of canna is forwarded in proliferated culture medium again and carries out Multiplying culture, finally the 2cm of length 1 differentiated Multiple Buds are transferred in root media and cultivates acquisition and takes root canna.The present invention is by being induced canna, being bred, being taken root, it is quick to obtain canna tissue-cultured seedling, establish good tissue culture rapid propagation system, take root fast, shorten the canna tissue culture time, cost is low, while improves canna rooting rate and survival rate, has preferable economic benefit, social benefit and ecological benefits.

Description

A kind of method of canna cultured in vitro
Technical field
The present invention relates to field of plant growing technology, is related to a kind of method of canna cultured in vitro.
Background technology
Canna (Canna indica), alias orchid any of several broadleaf plants, canna, for Cannaceae, canna platymiscium, perennial napiform root Herbage flower, originate in the ground such as american torrid zone and Africa.Canna plant height is grown thickly, blade is big, blade up to 100~180cm Various colors, Hua great, pattern is gorgeous, color is various, and florescence length, is a kind of very excellent afforestation, beautification of landscape plant. Canna blade can absorb the harmful substances such as sulfur dioxide, hydrogen chloride, though blade is vulnerable, can grow young leaves again, quickly Restoration ecosystem, monitoring pernicious gas pollution indicator plant therefore can be also used as, there is purification air, environmental protection effect.In addition, Canna is also used as the diseases such as drug therapy sore swollen toxin, uterine hemorrhage, leukorrhea, acute icterohepatitis.Have some cities now City is planted canna as main green plants, but because traditional modes of reproduction seeling industry speed is slow, serious limit Canna has been made to apply on a large scale.At present, the breeding of canna is mainly carried out by seed and stem tuber division propagation mode.But It is that the big kind of existing market demand substantially setting percentage is very low, even shaky, seed amount is limited, and passes through stem tuber plant division Mode is bred to be limited by material state itself, production season and soil, and annual output much can not meet the demand in market. Simultaneously as the long-term propagating materials of canna locates underground for a long time, easily by various viral infection, because canna uses more Vegetative propagation, once infection is just difficult to be eradicated by chemicals.In order to form canna large-scale production, explore a kind of suitable The method for closing canna tissue rapid propagation is necessary.
The content of the invention
The present invention is for overcome the deficiencies in the prior art, there is provided a kind of growth result is good, expands numerous fast canna tissue culture Propagation method.
In order to realize foregoing invention purpose, technical scheme is as follows：
A kind of method of canna cultured in vitro, including explant processing, Fiber differentiation, Multiplying culture and culture of rootage, choose Rhizome base portion with bud is cut into stem with bud, explant is washed by rhizome of the canna underground with bud as explant After net sterilization processing, it is inoculated in culture in inducing culture and obtains the initial bud of canna, then the initial bud of canna is transferred Multiplying culture is carried out into proliferated culture medium, finally the long 1-2cm differentiated Multiple Buds are transferred in root media and trained It is foster to obtain the canna that takes root；Mainly comprise the following steps：
（1）Explant processing：Rhizome of the canna underground with bud is chosen as explant, explant 2-3 is rinsed with pure water It is secondary, the rhizome base portion with bud is cut into stem with bud, then soaks 30- in the zineb blue powder solution of volumetric concentration 0.1% 40min, the % HgCl of volumetric concentration 0.15 are used after being cleaned with clear water2Solution carries out sterilization to explant immersion 15-20min, Finally with after the clean explant of aseptic water washing, it is placed on aseptic filter paper and blots water, it is standby；
（2）Fiber differentiation：By step（1）The stem section of rhizome of the canna underground with bud after processing is inoculated into inducing culture Middle culture, first light culture 3-4 d, then illumination cultivation, light application time 12h/d, intensity of illumination 500-1000lx, cultivation temperature 22-28 DEG C, incubation time 20-22d, culture obtains the initial bud of canna；
（3）Multiplying culture：By step（2）It is middle to induce the obtained initial bud of canna to be transferred in proliferated culture medium, light application time 10-12h/d, intensity of illumination 3000-3500lx, 22-28 DEG C of cultivation temperature, incubation time 20-25d, differentiation budding；
（4）Culture of rootage：By step（3）In bud length to the 1-2cm Multiple Buds that differentiate be transferred in root media, light According to time 12h/d, intensity of illumination 1500-2000lx, 22-28 DEG C of cultivation temperature, incubation time 20-30d, beauty of taking root is obtained Any of several broadleaf plants.
Above step（2）The formula of described inducing culture is：Improve LS culture medium+KT 6.0-8.0 mg/L+NAA 15 mg/L of 2.0-3.0 mg/L+VC+brassinosteroid 20mg/L+VB2 The g/ of 15 mg/+ agar powders, 3.6 g/L+ sucrose 20 L。
Above step（3）The formula of described proliferated culture medium is：Improve LS culture medium+KT 3.0-5.0 mg/L+NAA The g/L+ of 10 mg/L+ brassinosteroid 20mg/L+ agar powders of 1.5-2.0 mg/L+ZT 0.1-0.5 mg/L+ glycine 3.6 The g/L of sucrose 30.
Above step（4）The formula of described root media is：Improve LS culture medium+KT 1.5-2.0 mg/L+NAA 0.5-1.0 mg/L+VC 10 mg/L+VB2 The mg/L+ PVPK30 mg/L+ agar powders of 20 mg+ glycine 10 The g/L of 3.6 g/L+ sucrose 30.
The formula of above-described improvement LS culture mediums is：NH4NO3 880 mg/L+KNO3 780mg/L+CaCl2·2H2O 300mg/L+MgSO4.·7H2O 350 mg/L+KH2PO4 220 mg/L+H3BO3 4.5 mg/L+MnSO4.·4H2O 22.3 mg/L+ZnSO4·4H2O 8.6 mg/L+KI0.56mg/L+Na2MoO4·H2O 0.25 mg/L+CuSO4.·5H2O 0.025 mg/L+CoCl2·6H2O 0.025 mg/L+FeSO4The mg/L+ nicotinic acid of 27.8 mg/L+NaEDTA, 37.0 mg/L+ inositols 75 The .5 mg/L of 0.5 mg/L+ puridoxine hydrochlorides, 02 mg/L+ puridoxine hydrochlorides of .5 mg/L+ glycine 0.
The present invention has the advantage that and had the beneficial effect that：
1st, the present invention avoids progeny variation feelings caused by induction using rhizome of the canna underground with bud as explant Condition, by inducing culture culture, the initial bud induction rate 100% of canna；Access and take root after 4-5 cycle proliferation culture Cultivated in culture medium, rooting rate reaches 100%.
2nd, the present invention is improved to LS minimal mediums, while also adds many kinds of substance, and rapid effective suppression is beautiful The lesion of people's any of several broadleaf plants tissue culture bud, at the same promote bud grow and it is sturdy so that the nutrition of culture medium is more comprehensive, more effectively promotes The generation of canna bud induction, breed and take root, realize a large amount of propagation.
3rd, the present invention is quick to obtain canna tissue-cultured seedling by being induced canna, breeding, taking root, and establishes good Good tissue culture rapid propagation system, take root fast, shorten the canna tissue culture time, cost is low, while improve canna rooting rate and Survival rate, there are preferable economic benefit, social benefit and ecological benefits.
Embodiment
With reference to specific embodiment, the present invention will be further described.
Embodiment 1：
Rhizome of the canna underground with bud is chosen as explant, the rhizome base portion with bud is cut into stem with bud, use is pure Water rinses explant 2-3 times, then soaks 30min in the zineb blue powder solution of volumetric concentration 0.1%, is used after being cleaned with clear water The % HgCl of volumetric concentration 0.152Solution carries out sterilization to explant immersion 15-20min, finally clean with aseptic water washing After explant, it is placed on aseptic filter paper and blots water, it is standby.
The stem section of rhizome of the canna underground after processing with bud is inoculated into inducing culture and cultivated, first light culture 3-4 d, then illumination cultivation, light application time 12h/d, intensity of illumination 500lx, 22-25 DEG C of cultivation temperature, incubation time 20- 22d, culture obtain the initial bud of canna, and initial bud induction rate is up to 99%；Wherein, the formula of described inducing culture is：Improvement 3.0 mg/L+NAA of LS culture mediums+KT 2.0,15 mg/L of mg/L+VC+brassinosteroid 20mg/L+VB2 15 mg/+ fine jades The g/L of 3.6 g/L+ sucrose of cosmetics 20.
The initial bud of obtained canna is transferred in proliferated culture medium, light application time 10h/d, intensity of illumination 3000lx, 22-25 DEG C of cultivation temperature, incubation time 20-25d, differentiation budding；Wherein, the formula of described proliferated culture medium is：Improve LS The mg/L+ brassinosteroids 20mg/L+ of 0.1 mg/L+ glycine of culture medium+KT 3.0 mg/L+NAA, 1.5 mg/L+ZT 10 The g/L of 3.6 g/L+ sucrose of agar powder 30.
Bud length to the 1-2cm Multiple Buds that Proliferation, Differentiation goes out are transferred in root media, light application time 12h/d, light According to intensity 1500lx, 22-25 DEG C of cultivation temperature, incubation time 20-30d obtains the canna that takes root, and rooting rate reaches 100%；Its In, the formula of described root media is：Improve the mg/L+ of 0.5 mg/L+VC of LS culture mediums 1.5 mg/L+NAA of+KT 10 VB2 The g/L of 20 mg+ glycine, 10 3.6 g/L+ sucrose of mg/L+ PVPK30 mg/L+ agar powders 30.
The formula of above-described improvement LS culture mediums is：NH4NO3 880 mg/L+KNO3 780mg/L+CaCl2·2H2O 300mg/L+MgSO4.·7H2O 350 mg/L+KH2PO4 220 mg/L+H3BO3 4.5 mg/L+MnSO4.·4H2O 22.3 mg/L+ZnSO4·4H2O 8.6 mg/L+KI0.56mg/L+Na2MoO4·H2O 0.25 mg/L+CuSO4.·5H2O 0.025 mg/L+CoCl2·6H2O 0.025 mg/L+FeSO4The mg/L+ nicotinic acid of 27.8 mg/L+NaEDTA, 37.0 mg/L+ inositols 75 The .5 mg/L of 0.5 mg/L+ puridoxine hydrochlorides, 02 mg/L+ puridoxine hydrochlorides of .5 mg/L+ glycine 0.
Embodiment 2：
Rhizome of the canna underground with bud is chosen as explant, the rhizome base portion with bud is cut into stem with bud, use is pure Water rinses explant 2-3 times, then soaks 35min in the zineb blue powder solution of volumetric concentration 0.1%, is used after being cleaned with clear water The % HgCl of volumetric concentration 0.152Solution carries out sterilization to explant immersion 15-20min, finally clean with aseptic water washing After explant, it is placed on aseptic filter paper and blots water, it is standby.
The stem section of rhizome of the canna underground after processing with bud is inoculated into inducing culture and cultivated, first light culture 3-4 d, then illumination cultivation, light application time 12h/d, intensity of illumination 1000lx, 25-26 DEG C of cultivation temperature, incubation time 20- 22d, culture obtain the initial bud of canna, and initial bud induction rate is up to 99%；Wherein, the formula of described inducing culture is：Improvement 3.5 mg/L+NAA of LS culture mediums+KT 2.5,15 mg/L of mg/L+VC+brassinosteroid 20mg/L+VB2 15 mg/+ fine jades The g/L of 3.6 g/L+ sucrose of cosmetics 20.
The initial bud of obtained canna is transferred in proliferated culture medium, light application time 11h/d, intensity of illumination 3000lx, 25-26 DEG C of cultivation temperature, incubation time 20-25d, differentiation budding；Wherein, the formula of described proliferated culture medium is：Improve LS The mg/L+ brassinosteroids 20mg/L+ of 0.4 mg/L+ glycine of culture medium+KT 3.5 mg/L+NAA, 1.8 mg/L+ZT 10 The g/L of 3.6 g/L+ sucrose of agar powder 30.
Bud length to the 1-2cm Multiple Buds that Proliferation, Differentiation goes out are transferred in root media, light application time 12h/d, light According to intensity 200lx, 25-26 DEG C of cultivation temperature, incubation time 20-30d obtains the canna that takes root, and rooting rate reaches 100%；Its In, the formula of described root media is：Improve the mg/L+ of 0.8 mg/L+VC of LS culture medium+KT 1.8mg/L+NAA 10 VB2 The g/L of 20 mg+ glycine, 10 3.6 g/L+ sucrose of mg/L+ PVPK30 mg/L+ agar powders 30.
The formula of above-described improvement LS culture mediums is：NH4NO3 880 mg/L+KNO3 780mg/L+CaCl2·2H2O 300mg/L+MgSO4.·7H2O 350 mg/L+KH2PO4 220 mg/L+H3BO3 4.5 mg/L+MnSO4.·4H2O 22.3 mg/L+ZnSO4·4H2O 8.6 mg/L+KI0.56mg/L+Na2MoO4·H2O 0.25 mg/L+CuSO4.·5H2O 0.025 mg/L+CoCl2·6H2O 0.025 mg/L+FeSO427.8 the mg/L+ nicotinic acid of 37.0 mg/L+ inositols of mg/L+NaEDTA 75 The .5 mg/L of 0.5 mg/L+ puridoxine hydrochlorides, 02 mg/L+ puridoxine hydrochlorides of .5 mg/L+ glycine 0.
Embodiment 3：
Rhizome of the canna underground with bud is chosen as explant, the rhizome base portion with bud is cut into stem with bud, use is pure Water rinses explant 2-3 times, then soaks 40min in the zineb blue powder solution of volumetric concentration 0.1%, is used after being cleaned with clear water The % HgCl of volumetric concentration 0.152Solution carries out sterilization to explant immersion 15-20min, finally clean with aseptic water washing After explant, it is placed on aseptic filter paper and blots water, it is standby.
The stem section of rhizome of the canna underground after processing with bud is inoculated into inducing culture and cultivated, first light culture 3-4 d, then illumination cultivation, light application time 12h/d, intensity of illumination 1000lx, 26-28 DEG C of cultivation temperature, incubation time 20- 22d, culture obtain the initial bud of canna, and initial bud induction rate is up to 99%；Wherein, the formula of described inducing culture is：Improvement 4.0 mg/L+NAA of LS culture mediums+KT 3.0,15 mg/L of mg/L+VC+brassinosteroid 20mg/L+VB2 15 mg/+ fine jades The g/L of 3.6 g/L+ sucrose of cosmetics 20.
The initial bud of obtained canna is transferred in proliferated culture medium, light application time 12h/d, intensity of illumination 3500lx, 26-28 DEG C of cultivation temperature, incubation time 20-25d, differentiation budding；Wherein, the formula of described proliferated culture medium is：Improve LS The mg/L+ brassinosteroids 20mg/L+ of 0.5 mg/L+ glycine of culture medium+KT 5.0 mg/L+NAA, 2.0 mg/L+ZT 10 The g/L of 3.6 g/L+ sucrose of agar powder 30.
Bud length to the 1-2cm Multiple Buds that Proliferation, Differentiation goes out are transferred in root media, light application time 12h/d, light According to intensity 2000lx, 26-28 DEG C of cultivation temperature, incubation time 20-30d obtains the canna that takes root, and rooting rate reaches 100%；Its In, the formula of described root media is：Improve the mg/L+ of 1.0 mg/L+VC of LS culture mediums 2.0 mg/L+NAA of+KT 10 VB2 The g/L of 20 mg+ glycine, 10 3.6 g/L+ sucrose of mg/L+ PVPK30 mg/L+ agar powders 30.
The formula of above-described improvement LS culture mediums is：NH4NO3 880 mg/L+KNO3 780mg/L+CaCl2·2H2O 300mg/L+MgSO4.·7H2O 350 mg/L+KH2PO4 220 mg/L+H3BO3 4.5 mg/L+MnSO4.·4H2O 22.3 mg/L+ZnSO4·4H2O 8.6 mg/L+KI0.56mg/L+Na2MoO4·H2O 0.25 mg/L+CuSO4.·5H2O 0.025 mg/L+CoCl2·6H2O 0.025 mg/L+FeSO4The mg/L+ nicotinic acid of 27.8 mg/L+NaEDTA, 37.0 mg/L+ inositols 75 The .5 mg/L of 0.5 mg/L+ puridoxine hydrochlorides, 02 mg/L+ puridoxine hydrochlorides of .5 mg/L+ glycine 0.

Claims (5)

  1. A kind of 1. method of canna cultured in vitro, it is characterised in that：Including explant processing, Fiber differentiation, Multiplying culture and Culture of rootage, the rhizome of canna underground with bud is chosen as explant, the rhizome base portion with bud is cut into stem with bud, it is right Explant is carried out after cleaning sterilization processing, is inoculated in inducing culture culture and is obtained the initial bud of canna, then by beauty The initial bud of any of several broadleaf plants, which is forwarded in proliferated culture medium, carries out Multiplying culture, and the long 1-2cm differentiated Multiple Buds finally are transferred into life Acquisition is cultivated in root culture medium to take root canna；Mainly comprise the following steps：
    （1）Explant processing：Rhizome of the canna underground with bud is chosen as explant, the rhizome base portion with bud is cut into band Leaf stem section, rinse explant 2-3 times with pure water, then soak 30- in the zineb blue powder solution of volumetric concentration 0.1% 40min, the % HgCl of volumetric concentration 0.15 are used after being cleaned with clear water2Solution carries out sterilization to explant immersion 15-20min, Finally with after the clean explant of aseptic water washing, it is placed on aseptic filter paper and blots water, it is standby；
    （2）Fiber differentiation：By step（1）The stem section of rhizome of the canna underground with bud after processing is inoculated into inducing culture Middle culture, first light culture 3-4 d, then illumination cultivation, light application time 12h/d, intensity of illumination 500-1000lx, cultivation temperature 22-28 DEG C, incubation time 20-22d, culture obtains the initial bud of canna；
    （3）Multiplying culture：By step（2）It is middle to induce the obtained initial bud of canna to be transferred in proliferated culture medium, light application time 10-12h/d, intensity of illumination 3000-3500lx, 22-28 DEG C of cultivation temperature, incubation time 20-25d, differentiation budding；
    （4）Culture of rootage：By step（3）In bud length to the 1-2cm Multiple Buds that differentiate be transferred in root media, light According to time 12h/d, intensity of illumination 1500-2000lx, 22-28 DEG C of cultivation temperature, incubation time 20-30d, beauty of taking root is obtained Any of several broadleaf plants.
  2. A kind of 2. method of canna cultured in vitro according to claim 1, it is characterised in that：Step（2）Described lures The formula for leading culture medium is：The mg/L of improvement LS culture medium+KT 3.0-4.0 mg/L+NAA 2.0-3.0 mg/L+VC 15+ Brassinosteroid 20mg/L+VB2 The g/L of 15 mg/+ agar powders, 3.6 g/L+ sucrose 20.
  3. A kind of 3. method of canna cultured in vitro according to claim 1, it is characterised in that：Step（3）Described increasing The formula for growing culture medium is：Improve LS culture medium+KT 3.0-5.0 mg/L+NAA 1.5-2.0 mg/L+ZT 0.1-0.5 mg/ The g/L of 10 3.6 g/L+ sucrose of mg/L+ brassinosteroid 20mg/L+ agar powders of L+ glycine 30.
  4. A kind of 4. method of canna cultured in vitro according to claim 1, it is characterised in that：Step（4）Described life The formula of root culture medium is：Improve the mg/L+VB of LS culture medium+KT 1.5-2.0 mg/L+NAA 0.5-1.0 mg/L+VC 102 The g/L of 20 mg+ glycine, 10 3.6 g/L+ sucrose of mg/L+ PVPK30 mg/L+ agar powders 30.
  5. 5. according to a kind of method of any described canna cultured in vitro of claim 2-4, it is characterised in that：Described improvement The formula of LS culture mediums is：NH4NO3 880 mg/L+KNO3 780mg/L+CaCl2·2H2O 300mg/L+MgSO4.·7H2O 350 mg/L+KH2PO4 220 mg/L+H3BO3 4.5 mg/L+MnSO4.·4H2O 22.3 mg/L+ZnSO4·4H2O 8.6 mg/L+KI0.56mg/L+Na2MoO4·H2O 0.25 mg/L+CuSO4.·5H2O 0.025 mg/L+CoCl2·6H2O 0.025 mg/L+FeSO4The mg/L+ puridoxine hydrochlorides 0 of 27.8 mg/L+NaEDTA, 37.0 mg/L+ inositols, 75 mg/L+ nicotinic acid 0.5 .5 the .5 mg/L of 2 mg/L+ puridoxine hydrochlorides of mg/L+ glycine 0.
CN201711201037.0A 2017-11-27 2017-11-27 A kind of method of canna cultured in vitro Withdrawn CN107667868A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114766365A (en) * 2022-05-06 2022-07-22 贵州大学 Method for in-vitro germination and efficient rapid propagation of canna plant seeds

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114766365A (en) * 2022-05-06 2022-07-22 贵州大学 Method for in-vitro germination and efficient rapid propagation of canna plant seeds

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Application publication date: 20180209