CN101715730B - Tissue culture rapid propagation method for sugarcane new Taisugar No.10 - Google Patents
Tissue culture rapid propagation method for sugarcane new Taisugar No.10 Download PDFInfo
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Abstract
The invention relates to a tissue culture rapid propagation method for sugarcane new Taisugar No.10. The new Taisugar No.10 has high plant, compact plant shape, straight leaf, proper width, deep green leaves in the all growth stage and high yield increase potential, belongs to a variety of early mid-maturation, high yield and sugar and wide adaptability and is suitable for planting in a main planting area of China. At present, the sugarcane adopts stem bud asexual propagation and has low propagation coefficient, and the yield and the quality of sugarcane are greatly reduced caused by the repeated invasion of various disease source substances after planting for many years; the variety degradation of a fine variety is accelerated, the economic service life of the fine variety is shortened, and the rapid popularization and application of the sugarcane variety are greatly limited. The propagation method comprises the step of carrying out the propagation of cluster buds by axillary buds in sterile sugarcane sheaths and forming a complete plant by rhizogenesis culture. The axillary bud tissue culture rapid propagation technology of the new Taisugar No.10 is suitable for the industrial rapid propagation of a sugarcane seedling.
Description
Technical field:
The invention belongs to biotechnology engineering field, be specifically related to No. 10 tissue culture rapid propagation techniques of the new platform sugar of sugarcane.
Background technology:
No. 10 plant of the new platform sugar of sugarcane are tall and big, and plant type is compact, and big in the stem footpath, internode is longer, and leaf is straight and upright, and width is medium, and mesophyll is thicker, and the time of infertility, the leaf look dark green, and rudiment is fast, and tillering ability is strong and neat, and full phase growth is vigorous, and yield potential is big.Ripe in belonging to early, high yield, high sugar, degeneration-resistant strong, resistant to lodging, perennial root is strong, the kind of wide adaptability, the master who suits in China plants ground plantations such as Guangxi, district, Yunnan, Guangdong.The prior art sugarcane production adopts the stem eye vegetative propagation at present; Reproduction coefficient is low; And the decline significantly of sugarcane yield and quality is caused because of receiving infecting repeatedly of pathogeny thing in the plantation back for many years, causes breeding kind sexual involution; Breeding shortening service life has greatly limited the promotion and application of sugar cane breed.Tissue culture rapid propagation technique is that the quick breeding and the improved variety popularization of sugar cane breed provides effective means, and tissue culture technique not only can increase reproduction coefficient, saves big content of starting materials sugarcane, and remarkable to mosaic of sugarcane virus disease detoxification efficiency.Yet; The Different Crop kind is different to the reaction with a kind of medium; Have the varietY specificity problem, the culture effect that has is fine, and the then reproduction coefficient that has is very low; Some kind still can not form complete regenerating system, thus limited tissue culture technique sugarcane plants fast numerous with genetic improvement on application.Therefore, different sugar cane breeds need be inquired into different condition of tissue culture and method.No. 10, new platform sugar since introduction of plant, have sugar content height, branchs evil soon, advantages such as strong stress resistance, wide adaptability.But because provenance is few, price is expensive again, breeds popularization by the sugarcane stem of routine, the time is long, and cost is high.Adopting axillary bud tissue to cultivate method for quickly breeding is the new way of promoting this breeding rapidly.Do not see the report that No. 10 quick breeding method for tissue culture of the new platform sugar of sugarcane are arranged in the prior art.
Summary of the invention:
The present invention is intended to quicken with tissue culture rapid propagation technique the reproduction speed of No. 10, the new platform sugar of sugarcane, obtains a large amount of test-tube plantlets, for the needs that satisfy No. 10 seedling markets of the new platform sugar of sugarcane with improve sugarcane yield and sugar a valid approach be provided.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
No. 10 quick breeding method for tissue culture of the new platform sugar of sugarcane comprise the steps:
1) preparing culture medium: MS is as basal medium, additional saccharose 2.5-3.5%, agar powder 0.7-0.9%, pH value 5.6-5.8.Group training basic condition of culture of each stage is: 27 ± 2 ℃ of cultivation temperature, illuminance 1500LX, illumination every day 12 hours.Each component and every liter of contained weight of each stage medium of group training are following:
Axillalry bud propagating culture medium: MS adds 6-BA 1.5～3.0mg/L and NAA 0.1～0.3mg/L again as basal medium;
Bud proliferated culture medium: MS adds 6-BA 0.5～1.0mg/L and NAA 0.1mg/L again as basal medium;
Root media: 1/2MS adds the activated carbon of NAA 0.1～0.5mg/L and 0.05% again as basal medium.
2) select explant sterilization inoculation: choose No. 10 stem tips of the new platform sugar of sugarcane of 6-8 month, divest the outer leaf sheath of stem tip layer 2-3, get the long sugarcane stem of the following 40cm of stem apex; After alcohol with 70% is cleaned; Intercepting was soaked aseptic water washing 3-4 time from the sugarcane stem below the stem apex 10cm 15 minutes with 0.1% mercuric chloride+2 a tween thimerosal; Under aseptic condition, continue to divest outside leaf sheath; Cut the stem section of band axillalry bud again, be inoculated into the axillalry bud propagating culture medium and be that to add 6-BA 1.5～3.0mg/L among the MS last with NAA 0.1～0.3mg/L, the cultivation of inoculation back after 3～5 days axillalry bud grow;
3) hyperblastosis of liquid bud is cultivated: 2) axillalry bud to transfer to the shoot proliferation medium be to add among the MS 3 weeks of cultivation 6-BA0.5～1.0mg/L and NAA 0.1mg/L oned, axillalry bud differentiates many sugarcane seedlings formation clump buds;
4) culture of rootage is with 3) the clump bud that forms of propagation is cut into individual plant to be seeded in root media is to add among the 1/2MS in the activated carbon of NAA0.1～0.5mg/L and 0.05% to carry out 3 weeks of culture of rootage;
5) tissue cultivating seedling is transplanted: with 4) the gained tissue cultivating seedling is placed on 80% the greenhouse refining seedling 10 days of shading; Opened bottle cap refining seedling again 2～3 days; Clean medium then and be transplanted on the nursery matrix of loam, peat soil and river sand preparation in 1: 1: 1 by volume, heel in 45 days and go out transplanting land for growing field crops, garden.
The present invention more specifically scheme can be summarized as:
1) axillalry bud breeding is cultivated: choose 6-8 month stalwartness of sugarcane field growth, the good cane stalk tip of proterties is as explant.Get the long sugarcane stem from the following 40cm of stem apex, after the alcohol with 70% was cleaned 1 time, the sugarcane stem below the intercepting stem apex 10cm soaked 15 minutes with 0.1% mercuric chloride+2 a tween thimerosal again.Aseptic water washing 3-4 time cuts the stem section that is about 2cm band axillalry bud again, is inoculated on the axillary bud tissue medium (2).Remaining part is peeled Bao Ye off, directly downcuts to have the stem section of axillalry bud, is inoculated on the identical medium and cultivates.The axillalry bud elongation is up to 3cm after 3～5 days.
2) shoot proliferation is cultivated: will cut off the axillalry bud of elongation from base portion, and insert on the medium (3), after 5 days, base portion grows many small salient points, after 10 days, differentiates many budlets, forms the bud clump.Constantly cutting bud clump obtains a large amount of tissue cultivating seedling soon.With 2 weeks be 1 subculture cycle, each cycle can be bred the new talent more than 2 times.
3) inducing of root: connect subculture seedling (being no more than for 9 generations) on the root media (4), base section dissolves many adventive root after 5 days, grows many radial after 10 days, goes out the root rate and reaches more than 80%.
4) refining seedling and transplanting: rooting tube plantlet is placed outdoor sunlight lower refining seedling; Remove bottle cap after 10 days; Refined again 3 days; Taking-up is cleaned medium with running water, immerses in 1000 times the 70% thiophanate methyl solution 10 minutes, plants new loam+peat soil+sandy soil by in 1: 1: 1 mixed matrix.Whenever sprayed water 1 time at a distance from 3 days, just portable is to the land for growing field crops after 45 days, and survival rate reaches more than 90%, and the output of tissue cultivating seedling is identical with the kind stem breeding.
The inventive method comprises the explant of selecting suitable position, optimizes the axillary bud tissue medium, shoot proliferation medium and root media and high-servival rate cultivation matrix.Make the sugarcane axillary bud tissue cultivate and allly fast plant simplely that strong operability is promoted the use of easily.Can accelerate the sugarcane reproduction speed greatly and shorten the production cycle, be applicable to cane seedling production and breeding work.Enforcement should technology, only needs conventional Plant Tissue Breeding equipment to carry out, and has less investment, and cost is low, and the characteristics that output is high are suitable for the cane seedling batch production and breed fast.
Embodiment:
Further specify essentiality content of the present invention with embodiments of the invention below, but do not limit the present invention with this.
Embodiment 1:
1) axillalry bud breeding is cultivated: choose 6-8 month stalwartness of sugarcane field growth, the good cane stalk tip of proterties is as explant.Get the long sugarcane stem of the following 40cm of stem apex, the alcohol with 70% again from the sugarcane stem below its intercepting 10cm, soaked 15 minutes with 0.1% mercuric chloride+2 a tween thimerosal after cleaning 1 time.Aseptic water washing 3-4 time cuts the stem section that is about 2cm band axillalry bud again, is inoculated on the axillary bud tissue medium (2).Remaining part is peelled off all the other leaf sheaths, and near the bud of each stipes of stem apex is all come out; Downcut the internode conduct that only has an axillalry bud on the stem section one by one and grow body outward; Be inoculated into the axillalry bud propagating culture medium and be add 6-BA 1.5～3.0mg/L among the MS and NAA 0.1～0.3mg/L last, be placed on 27 ± 2 ℃ of temperature after the inoculation, illumination 1500LX; Illumination every day was cultivated under the condition in 12 hours, and the axillalry bud elongation is up to 3cm after 3～5 days.
2) shoot proliferation is cultivated: the axillalry bud that will cut off elongation from base portion; Inserting the shoot proliferation medium is to add on 6-BA 0.5～1.0mg/L and the NAA 0.1mg/L among the MS to cultivate, and after 5 days, base portion grows many small salient points; Differentiate many budlets after 10 days, form the clump bud.Constantly cutting clump bud obtains a large amount of tissue cultivating seedling soon.With 2 weeks be 1 subculture cycle, each cycle can be bred the new talent more than 2 times.
3) inducing of root: connecing root media into to subculture seedling (being no more than for 9 generations) is to add among the 1/2MS in the activated carbon of NAA0.1～0.5mg/L and 0.05% to carry out culture of rootage; Base section dissolves many adventive root after 5 days; Grow many radial after 10 days, go out the root rate and reach more than 80%.
4) refining seedling and transplanting: with 3) the gained rooting tube plantlet places 80% the greenhouse refining seedling 10 days of shading; Remove bottle cap after 10 days; Refined again 3 days; Taking-up is cleaned medium with running water, immerses in 1000 times the 70% thiophanate methyl solution 10 minutes, plants in 1: 1: 1 by volume matrix prepared of new loam+peat soil+sandy soil.Whenever sprayed water 1 time at a distance from 3 days, just portable is to the land for growing field crops after 45 days, and survival rate reaches more than 90%, and the output of tissue cultivating seedling is identical with the kind stem breeding.
Claims (2)
1. No. 10 quick breeding method for tissue culture of the new platform sugar of sugarcane comprise the steps:
1) preparing culture medium, minimal medium with each component and every liter of contained weight of other group training stage medium is:
Basal medium: MS is as basal medium, additional saccharose 2.53.5%, agar powder 0.7-0.9%, pH value 5.6-5.8,27 ± 2 ℃ of cultivation temperature, illuminance 1500LX, illumination 12 hours;
Axillalry bud propagating culture medium: add 6-BA 1.5～3.0mg/L and NAA 0.1～0.3mg/L among the MS;
Bud proliferated culture medium: add 6-BA 0.5～1.0mg/L and NAA 0.1mg/L among the MS;
Root media: the activated carbon that adds NAA 0.1～0.5mg/L and 0.05% among the 1/2MS;
2) select explant sterilization inoculation: choose No. 10 stem tips of the new platform sugar of sugarcane of 6-8 month, divest the outer leaf sheath of stem tip layer 2-3, get the long sugarcane stem of the following 40cm of stem apex; After alcohol with 70% was cleaned, the sugarcane stem below the intercepting 10cm added 2 tween thimerosals with 0.1% mercuric chloride and soaked 15 minutes; Aseptic water washing 3-4 time continues to divest outside leaf sheath under aseptic condition, cut the stem section of the band axillalry bud that is about 2cm again; Be inoculated into the axillalry bud propagating culture medium and be add 6-BA 1.5～3.0mg/L among the MS and NAA 0.1～0.3mg/L last, be placed on 27 ± 2 ℃ of temperature after the inoculation, light intensity 1500LX; Illumination was cultivated under the condition in 12 hours, and axillalry bud grows after 3～5 days;
3) axillary bud tissue enrichment culture: 2) breeding obtains axillalry bud to transfer to the shoot proliferation medium is to add 6-BA 0.5～1.0mg/L among the MS to go up with NAA 0.1mg/L and cultivated for 3 weeks, and callus differentiates the sugarcane seedling, formation clump bud;
4) culture of rootage is with 3) the clump bud that forms of propagation is cut into individual plant to be seeded in root media is to add among the 1/2MS in the activated carbon of NAA0.1～0.5mg/L and 0.05% to carry out 3 weeks of culture of rootage;
5) tissue cultivating seedling is transplanted: with 4) the gained tissue cultivating seedling is placed on 80% the greenhouse refining seedling 10 days of shading; Remove bottle cap after 10 days; Refined again 3 days, and be transplanted to then on the nursery matrix of loam+peat soil+river sand preparation in 1: 1: 1 by volume, heel in 45 days and go out transplanting land for growing field crops, garden.
2. method according to claim 1 is characterized in that: said minimal medium with group each each component of stage medium of training and every liter of contained weight is:
Basal medium: MS is as basal medium, additional saccharose 2.5-3.5%, agar powder 0.7-0.9%, pH value 5.6-5.8,27 ± 2 ℃ of cultivation temperature, illuminance 1500LX, illumination 12 hours;
Axillalry bud propagating culture medium: add 6-BA 3.0mg/L and NAA 0.3mg/L among the MS;
Bud proliferated culture medium: add 6-BA 1.0mg/L and NAA 0.1mg/L among the MS;
Root media: the activated carbon that adds NAA 0.5mg/L and 0.05% among the 1/2MS.
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| CN2009102183858A CN101715730B (en) | 2009-12-17 | 2009-12-17 | Tissue culture rapid propagation method for sugarcane new Taisugar No.10 |
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Families Citing this family (7)
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| CN102823493B (en) * | 2012-08-20 | 2013-07-17 | 云南云蔗科技开发有限公司 | Method of sugarcane distant hybrid embryo in-vitro culture and seedling breeding |
| CN102907328B (en) * | 2012-11-14 | 2014-08-27 | 中国热带农业科学院热带生物技术研究所 | Method for utilizing microelements to accelerate tissue culture seedling of sugarcane |
| CN105684902A (en) * | 2016-01-28 | 2016-06-22 | 广西南亚热带农业科学研究所 | Industrialized seedling culture method for detoxified health sugarcane seedlings |
| CN105875409B (en) * | 2016-04-13 | 2018-05-29 | 广东省农业科学院作物研究所 | A kind of acquisition methods of sugarcane aseptic explant |
| CN106234069B (en) * | 2016-08-08 | 2019-12-03 | 云南省农业科学院甘蔗研究所 | A kind of method of the peat soil matrix and its seedbed preparation of culturing sugarcane seedlings |
| CN107094623A (en) * | 2017-04-19 | 2017-08-29 | 江苏农林职业技术学院 | A kind of propagation method of sugarcane tissue culture |
| CN113349056A (en) * | 2021-07-07 | 2021-09-07 | 江门粤恬科技有限公司 | Tissue culture rapid propagation method for detoxification of black skin chewing cane |
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