CN106605595A - Method for breeding improved variety of hemp through shoot tip - Google Patents

Method for breeding improved variety of hemp through shoot tip Download PDF

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Publication number
CN106605595A
CN106605595A CN201611140039.9A CN201611140039A CN106605595A CN 106605595 A CN106605595 A CN 106605595A CN 201611140039 A CN201611140039 A CN 201611140039A CN 106605595 A CN106605595 A CN 106605595A
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weight portion
seedling
hemp
plant
stem apex
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CN201611140039.9A
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CN106605595B (en
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罗永志
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Qinzhou Qinnan Jinwo Industrial Park Management Committee
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罗永志
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/001Culture apparatus for tissue culture
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008Methods 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)
  • Cultivation Of Plants (AREA)

Abstract

The invention discloses a method for breeding an improved variety of hemp through a shoot tip, belonging to the technical field of hemp plantation. The method comprises the following steps: plant screening, pretreatment and sterilization of cut embryo buds, cutting of leaf primordial, cultivation of tissue culture seedlings, domestication of the seedlings, transplanting of the seedlings, etc. The method provided by the invention overcomes the problem of great difficulty in large-scale production of hemp caused by low seed germination rate, long germination acceleration time, inconsistent germination time, difficulty in collection of hemp seed resources, incapability of remaining all the excellent properties of male parents and female parents due to variation of filial generations and the like in traditional seed propagation of hemp.

Description

Cultivated fine seed strains by stem apex the method for Urtica cannabina L.
Technical field
The present invention relates to Urtica cannabina L. planting technology field, especially a kind of method of Urtica cannabina L. of being cultivated fine seed strains by stem apex.
Background technology
Urtica cannabina L. sapling multiplication technology is mainly that based on sexual propagation method, the advantage of seminal propagation is once to broadcast with seminal propagation Planting can obtain a large amount of nursery stocks, seed collection, storage, convenient transportation, and seedling growth is vigorous, strong stress resistance, easily domestication.But pass There is following defect in the Urtica cannabina L. seminal propagation of system:(1) percentage of seedgermination is low, and highest only reaches 80% or so;(2) germination time is long Up to 20 days or so, and germinating time was inconsistent;(3) Urtica cannabina L. resource scarcity, batch capture is difficult;(4)Syngenesis produce variation Probability it is high, filial generation possibly cannot retain the maternal whole good characteristics of male parent.The technology of Urtica cannabina L. seminal propagation at present is present Problem seriously hinders the large-scale production of Urtica cannabina L., thus cannot meet demand of the market to Urtica cannabina L., it is necessary to which setting up can be a large amount of, fast The method of speed breeding Urtica cannabina L. seedling.
Plant cell has totipotency, i.e. each plant cell containing all gene that can produce whole plant.It is theoretical On say, as long as condition is suitable, containing all genetic cells can Differentiation, produce whole plant.Mitogenetic group of stem apex It is usually virion that is nontoxic or only containing extremely low concentration in knitting, and in older tissue, viral level is with a distance from stem apex Increase and improve.Accordingly principle can obtain nontoxic plant using stem tip tissue culture.Using this principle, Urtica cannabina L. can be carried out good Plant and breed, cultivate virus-free seedling.
The content of the invention
The present invention provides a kind of method of Urtica cannabina L. of cultivating fine seed strains by stem apex, in the way of it can be to solve seminal propagation Urtica cannabina L. Cannot large-scale production breeding Urtica cannabina L. problem.
In order to solve the above problems, the technical solution adopted in the present invention is:It is this to be cultivated fine seed strains Urtica cannabina L. by stem apex Method, comprises the steps:
A, plant screening:When Urtica cannabina L. is ripe or during rudiment, the plant of no disease and pests harm is selected;
B, pretreatment and sterilization:Complete sub- bud of the length of step A gained plant more than 3cm is taken, it is clean with clean water, then use 70% ethanol rinsing 30s, then with 0.1% HgCl2Solution sterilization 3min, aseptic washing 3 is throughout the above;
C, the sub- bud obtained by step B is placed under microscope, after alcohol-pickled, then the tweezers that cooled down with calcination and shears are by son Bud meristematic zone cell is cut into the stem apex with 1~2 phyllopodium, then with sterile water wash 3 throughout the above;
D, cultivation:Step C gained stem apex segment is put into and is lured containing MS+6- benzyl purines 1.5mg/L+naphthalene acetic acid 1.5mg/L Lead and cultivate in culture medium to 3cm~4cm Multiple Buds are grown, then the Multiple Buds are cut into into 3 leaves, be about the segment of 1.5cm Access MS full dose proliferated culture mediums, 7 days~10 days be 1 switching cycle, propagation 3 cycles after, seedling is accessed in batches containing The root media culture of MS+6- benzyl purines 1.5mg/L+naphthalene acetic acid 1.0mg/L is grown to there is white root system;
E, the seedling of long root that step D gained comes into leaves are put into 3~5 days under sunlight, take out seedling, are transplanted to seedling bed, OK Away from 5~6cm, 5~6cm of spacing in the rows, deep 3~4cm is opened, one plant of seedling of often place's plantation keeps seedling bed moistening;The seedling bed be by Walnut husk is mixed with cane mill's filter mud and/or fish pond mud, calcium magnesium phosphate ferments and obtain as substrate;As 60% children When Seedling plant length is to height of seedling 50cm~60cm, land for growing field crops is transplanted to, you can.
In above-mentioned technical proposal, more specifically technical scheme can also be:The condition of culture of the breeding phase is temperature 23~28 DEG C, 1500~1800LUX of intensity of illumination.
Further:Each composition proportion of the seedling bed is weight portion~100 weight portion of walnut husk 80, cane mill Filter mud and/or weight portion~25 weight portion of fish pond mud 20, weight portion~10 weight portion of dipotassium hydrogen phosphate 5, the weight of magnesium sulfate 5 Part~10 weight portions, weight portion~10 weight portion of zinc sulfate 5.
As a result of above-mentioned technical proposal, the present invention has the advantages that compared with prior art:
1st, the present invention cultivates Urtica cannabina L. by the asexual reproduction method of stem apex, and the mode for solving traditional seminal propagation Urtica cannabina L. is brought Percentage of seedgermination is low, germination time length and germinating time is inconsistent, collection Urtica cannabina L. seed resource is difficult, filial generation is due to variation The big problem of large-scale production Urtica cannabina L. difficulty caused by maternal whole merit of male parent etc. cannot be retained.
2nd, the present invention can cultivate virus-free breeding Urtica cannabina L. seedling using the cellular omnipotency of stem apex, be planted Plant, plant strain growth is healthy and strong, growing way is consistent, and pest and disease damage incidence rate is low, and can keep all premium properties of wild Urtica cannabina L. and effective Component content.
Specific embodiment
Below in conjunction with specific embodiment, the invention will be further described:
The method of this Urtica cannabina L. of being cultivated fine seed strains by stem apex, is comprised the steps:
A, plant screening:When Urtica cannabina L. is ripe or during rudiment, the plant of no disease and pests harm is selected;
B, pretreatment and sterilization:Complete sub- bud of the length of step A gained plant more than 3cm is taken, it is clean with clean water, then use 70% ethanol rinsing 30s, then with 0.1% HgCl2Solution sterilization 3min, aseptic washing 3 is throughout the above;
C, the sub- bud obtained by step B is placed under microscope, after alcohol-pickled, then the tweezers that cooled down with calcination and shears cut Stem apex with 1~2 phyllopodium, then with sterile water wash 3 throughout the above;
D, cultivation:Or step C gained stem apex segment is put into containing MS+6- benzyl purines 1.5mg/L+naphthalene acetic acid 1.5mg/ Cultivate in L inducing cultures to 3cm~4cm Multiple Buds are grown, then the Multiple Buds are cut into into 3 leaves, be about the little of 1.5cm Section accesses MS full dose proliferated culture mediums, is within 7 days 1 switching cycle, after 3 cycles of propagation, seedling is accessed containing MS+6- in batches The root media culture of benzyl purine 1.5mg/L+naphthalene acetic acid 1.0mg/L is grown to there is white root system;
Above condition of culture be 23~28 DEG C of temperature, 1500~1800LUX of intensity of illumination;
E, the seedling of long root that step D gained comes into leaves are put into 3~5 days under sunlight, take out seedling, are transplanted to seedling bed, OK Away from 5~6cm, 5~6cm of spacing in the rows, deep 3~4cm is opened, one plant of seedling of often place's plantation keeps seedling bed moistening;The seedling bed be by The weight portion of walnut husk 80 is mixed with the weight portion of cane mill's filter mud 10, the weight portion of fish pond mud 10, phosphoric acid hydrogen two as substrate The weight portion of potassium 5, the weight portion of magnesium sulfate 5, the weight portion of zinc sulfate 5 ferment and obtain;When 60% seedling plants length to height of seedling 50cm~ During 60cm, land for growing field crops is transplanted to, you can.

Claims (3)

1. a kind of method of Urtica cannabina L. of being cultivated fine seed strains by stem apex, it is characterised in that comprise the steps:
A, plant screening:When Urtica cannabina L. is ripe or during rudiment, the plant of no disease and pests harm is selected;
B, pretreatment and sterilization:Complete sub- bud of the length of step A gained plant more than 3cm is taken, it is clean with clean water, then use 70% ethanol rinsing 30s, then with 0.1% HgCl2Solution sterilization 3min, aseptic washing 3 is throughout the above;
C, the sub- bud obtained by step B is placed under microscope, after alcohol-pickled, then the tweezers that cooled down with calcination and shears are by son Bud meristematic zone cell is cut into the stem apex with 1~2 phyllopodium, then with sterile water wash 3 throughout the above;
D, cultivation:Step C gained stem apex segment is put into and is lured containing MS+6- benzyl purines 1.5mg/L+naphthalene acetic acid 1.5mg/L Lead and cultivate in culture medium to 3cm~4cm Multiple Buds are grown, then the Multiple Buds are cut into into 3 leaves, be about the segment of 1.5cm Access MS full dose proliferated culture mediums, 7 days~10 days be 1 switching cycle, propagation 3 cycles after, seedling is accessed in batches containing The root media culture of MS+6- benzyl purines 1.5mg/L+naphthalene acetic acid 1.0mg/L is grown to there is white root system;
E, the seedling of long root that step D gained comes into leaves are put into 3~5 days under sunlight, take out seedling, are transplanted to seedling bed, OK Away from 5~6cm, 5~6cm of spacing in the rows, deep 3~4cm is opened, one plant of seedling of often place's plantation keeps seedling bed moistening;The seedling bed be by Walnut husk is mixed with cane mill's filter mud and/or fish pond mud, calcium magnesium phosphate ferments and obtain as substrate;As 60% children When Seedling plant length is to height of seedling 50cm~60cm, land for growing field crops is transplanted to, you can.
2. the method for Urtica cannabina L. of being cultivated fine seed strains by stem apex according to claim 1, it is characterised in that:The breeding phase Condition of culture be 23~28 DEG C of temperature, 1500~1800LUX of intensity of illumination.
3. the method for Urtica cannabina L. of being cultivated fine seed strains by stem apex according to claim 1, it is characterised in that:The seedling bed it is each Composition proportion is weight portion~100 weight portion of walnut husk 80, cane mill's filter mud and/or weight portion~25 of fish pond mud 20 Weight portion, weight portion~10 weight portion of dipotassium hydrogen phosphate 5, weight portion~10 weight portion of magnesium sulfate 5, weight portion~10 of zinc sulfate 5 Weight portion.
CN201611140039.9A 2016-12-12 2016-12-12 It is cultivated fine seed strains the method for fiery fiber crops by stem apex Active CN106605595B (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10477791B2 (en) 2017-10-25 2019-11-19 Cell Science Holding Ltd. Method of production of phytocannabinoids for use in medical treatments

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1887043A (en) * 2006-07-13 2007-01-03 江苏省中国科学院植物研究所 Fast propagation method of cannabis for inductrial use
CN104782502A (en) * 2015-05-12 2015-07-22 中国农业科学院麻类研究所 Method for rapidly obtaining regenerated plants of fiber hemps

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1887043A (en) * 2006-07-13 2007-01-03 江苏省中国科学院植物研究所 Fast propagation method of cannabis for inductrial use
CN104782502A (en) * 2015-05-12 2015-07-22 中国农业科学院麻类研究所 Method for rapidly obtaining regenerated plants of fiber hemps

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
REN WANG ET.AL.,: "A MICROPROPAGATION SYSTEM FOR CLONING OF HEMP (CANNABIS SATIVA L.) BY SHOOT TIP CULTURE", 《PAK. J. BOT.,》 *
尹品训等: "大麻组织培养中玻璃化苗研究初报", 《云南农业科技》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10477791B2 (en) 2017-10-25 2019-11-19 Cell Science Holding Ltd. Method of production of phytocannabinoids for use in medical treatments
US11477953B2 (en) 2017-10-25 2022-10-25 Cell Science Holding Ltd. Method of production of phytocannabinoids for use in medical treatments
US11690335B2 (en) 2017-10-25 2023-07-04 Cell Science Holding Ltd Method of production of phytocannabinoids for use in medical treatments

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Address after: 535000 3 / F, building 1, Qinzhou nixing pottery cultural and Creative Industrial Park (Millennium ancient pottery city), Qinzhou City, Guangxi Zhuang Autonomous Region

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Address before: Fengshan County town the Guangxi Zhuang Autonomous Region Hechi city Wenchang 547600 Phoenix Road No. 22 South Lane

Patentee before: Luo Yongzhi