CN102550417B - Cultivation of Hangzhou white chrysanthemum polyploidy good varieties and one-step seedling rapid propagation technology - Google Patents
Cultivation of Hangzhou white chrysanthemum polyploidy good varieties and one-step seedling rapid propagation technology Download PDFInfo
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- CN102550417B CN102550417B CN201210008065.1A CN201210008065A CN102550417B CN 102550417 B CN102550417 B CN 102550417B CN 201210008065 A CN201210008065 A CN 201210008065A CN 102550417 B CN102550417 B CN 102550417B
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Abstract
Cultivation of Hangzhou white chrysanthemum polyploidy good varieties and a one-step seedling rapid propagation technology comprise the steps of performing virus-free treatment to Hangzhou white chrysanthemum buds with viruses, taking out and placing growing points on an optimized cell starting culture medium to be cultured for 40 days to obtain virus-free calluses and obtaining a large amount of cluster buds through many-time transplantation. The polyploid inducing efficiency of the Hangzhou white chrysanthemum cluster buds transplanted in the culture medium after the cluster buds are treated by 0.2% of colchicine solution for 36 hours is the highest and reaches to 13.3%. The invention provides the one-step propagation technology, test-tube plantlets are inoculated on the one-step culture medium only in one time, and two culturing processes including propagation and rooting can be finished within 30 days. By applying the one-step propagation technology, 50% of production cost can be reduced, simultaneously the propagation speed is further accelerated in time, the cultivation of the Hangzhou white chrysanthemum polyploidy good varieties has good application prospect in the production, and economic benefit of the Hangzhou white chrysanthemum polyploidy good varieties in popularization and application is further improved.
Description
Technical field
The present invention relates to a kind of agro-ecology new technical field, relate in particular to a kind of cultivation and forming seedling through one step culture fast breeding technique of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds.
Background technology
Chrysanthemum is the dry capitulum of feverfew chrysanthemum Chrysanthemum morifolium Ramat.Chrysanthemum is used as medicine with a long history, for conventional large traditional Chinese medicinal materials assortment, is again medicine-food two-purpose simultaneously, domestic and international market demand is increasing, and four medicinal your name chrysanthemums have FLOS CHRYSANTHEMI ALBA from Haizhou of China, tribute chrysanthemum, Chu chrysanthemum and milli chrysanthemum at present, are distributed on both sides of the Changjiang River, the output of FLOS CHRYSANTHEMI ALBA from Haizhou of China wherein, market demand is maximum.But FLOS CHRYSANTHEMI ALBA from Haizhou of China deterioration of variety is at present serious, for solving Dendranthema morifolium Varieties degenerate problem, improve output and the quality of chrysanthemum medicinal material, we apply tissue culture technique and first FLOS CHRYSANTHEMI ALBA from Haizhou of China have been carried out the research of Shoot Tip Culture virus-free technology, then at aseptic in vitro, induction and the seed selection research of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid have been carried out, and the yield traits of the strain that induction is obtained, chemical composition content and some physical signs are measured, the unremitting effort that it's 8 years is past anteroposterior diameter, the FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds of high yield and high quality have finally been selected, simultaneously in order to accelerate the production application popularization of improved seeds, studied successfully and induced FLOS CHRYSANTHEMI ALBA from Haizhou of China to produce the medium of a large amount of Multiple Buds, studied successfully test-tube plantlet one-step method cultivation technology, large scale application for FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds, reduce production costs, increase economic efficiency the feasibility of technology is provided.
Summary of the invention
This patent comprises the technological innovation of following 4 aspects:
1. FLOS CHRYSANTHEMI ALBA from Haizhou of China improved hot is processed virus-free technology
FLOS CHRYSANTHEMI ALBA from Haizhou of China is medicine-food two-purpose economic crops, and various places development is very fast, but FLOS CHRYSANTHEMI ALBA from Haizhou of China is easy to infect multiple virus disease, infected the FLOS CHRYSANTHEMI ALBA from Haizhou of China deterioration of variety of virus disease, leaf shrinkage, poor growth, flower is year by year little, lopsided, poor quality, general underproduction 30-80%.And increase the weight of year by year.The economic benefit of FLOS CHRYSANTHEMI ALBA from Haizhou of China significantly declines, and has seriously hindered the development that FLOS CHRYSANTHEMI ALBA from Haizhou of China is produced.At present to virus disease also effective agricultural chemicals useless prevent and treat, even if there are some agricultural chemicals to use, but virus disease control poor effect also often causes that residue of pesticide exceed standard, virus disease has become " cancer " that FLOS CHRYSANTHEMI ALBA from Haizhou of China is produced.
For FLOS CHRYSANTHEMI ALBA from Haizhou of China virus disease, we have studied successfully first technology of this patent---and FLOS CHRYSANTHEMI ALBA from Haizhou of China improved hot is processed virus-free technology; Application the art of this patent virus-free effect can reach more than 90%, the detoxification FLOS CHRYSANTHEMI ALBA from Haizhou of China cultivating all shows obvious effect of increasing production in the production test of various places, effect of increasing production reaches 30-50% left and right, and general FLOS CHRYSANTHEMI ALBA from Haizhou of China Influenza Virus is more serious, and the effect of increasing production after application virus-free seedling is more obvious.
2. the induction of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid and breeding technique
On the successful basis of FLOS CHRYSANTHEMI ALBA from Haizhou of China virus-free technology, in order further to improve output and the quality of FLOS CHRYSANTHEMI ALBA from Haizhou of China, carried out cultivating the research of FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds, we have carried out induction and the seed selection of the FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid under aseptic condition, the 2nd technology--induction and the breeding technique of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid of the art of this patent have been formed, result of study shows: chrysanthemum accesses the most effective of induction polyploid in medium after 0.2% colchicine solution is processed 36h from sprouting, reach 13.3%.By root tip chromosomes Microscopic Identification; Field trial seed selection by 3 years is worked, and has successfully selected high yield, the FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds of high-quality, and output can further improve 30-50% than dliploid virus-free seedling.Can provide and produce upper Demonstration And Extension application.
3. induction FLOS CHRYSANTHEMI ALBA from Haizhou of China produces a large amount of culture medium prescriptions from sprouting
FLOS CHRYSANTHEMI ALBA from Haizhou of China virus-free, after polyploid improved seeds are cultivated successfully, in order to apply as early as possible on producing, need to solve the technical problem of the Fast-propagation of improved seeds seedling, can breed at short notice just a large amount of high quality seedlings provides in production and applies, and through a large amount of Screening of Media tests, has obtained a large amount of inducing clumping bud medium of FLOS CHRYSANTHEMI ALBA from Haizhou of China test-tube plantlet for this reason, obtained best FLOS CHRYSANTHEMI ALBA from Haizhou of China propagating culture medium formula, (MS+BA0.2-1.0mg/L+IAA0.05-0.5mg/L); Apply this culture medium prescription, monthly the reproduction speed of FLOS CHRYSANTHEMI ALBA from Haizhou of China can reach 20-30 doubly, for FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds large area, produces and lays a good foundation.
4. test-tube plantlet one-step method is cultivated technology
On the basis of above research work, FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds test-tube plantlet can reach in laboratory the batch production level of production, but each strain test-tube plantlet still needs to cultivate through 2-3 time, through 2-3 month cultivate one's ability, enough produce, in order further to reduce costs, improve the feasibility of production application, improve the economic benefit that FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds are applied, we have further carried out the short form test of medium, can discussion be carried out one-step method (an inoculated and cultured technology) Fast-propagation and go out a large amount of FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds test-tube plantlets, through lot of experiments, finally set up one-step method culture technique, test-tube plantlet is only by being once seeded in (MS+IAA0.2-1.0mg/L+PP333 0.1-1.0mg/L) on one-step method medium, just can in 30 days, successively complete breeding and two incubation of taking root.Application one-step method culture technique, can reduce production costs 50%, has further accelerated in time reproduction speed simultaneously, has aborning good application prospect, will further improve the economic benefit that FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds are applied.
Beneficial effect of the present invention: for FLOS CHRYSANTHEMI ALBA from Haizhou of China virus disease, we have studied successfully first technology of this patent---FLOS CHRYSANTHEMI ALBA from Haizhou of China improved hot is processed virus-free technology; Application the art of this patent virus-free effect can reach more than 90%, the detoxification FLOS CHRYSANTHEMI ALBA from Haizhou of China cultivating all shows obvious effect of increasing production in the production test of various places, effect of increasing production reaches 30-50% left and right, and general FLOS CHRYSANTHEMI ALBA from Haizhou of China Influenza Virus is more serious, and the effect of increasing production after application virus-free seedling is more obvious.The 2nd technology--induction and the breeding technique of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid of the art of this patent, the most effective of induction polyploid reaches 13.3%.And obtained best FLOS CHRYSANTHEMI ALBA from Haizhou of China propagating culture medium formula, and apply this culture medium prescription, monthly the reproduction speed of FLOS CHRYSANTHEMI ALBA from Haizhou of China can reach 20-30 doubly, applies one-step method culture technique simultaneously, can reduce production costs 50%.
Accompanying drawing explanation
Fig. 1 is the chromosome 2n=2x=40 of dliploid FLOS CHRYSANTHEMI ALBA from Haizhou of China of the present invention
Fig. 2 is the chromosome 2n=4x=80 of tetraploid FLOS CHRYSANTHEMI ALBA from Haizhou of China of the present invention
Fig. 3 is colchicine processing time and the effect test data statistics figure that the present invention induces FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid
Embodiment
The techniqueflow of this patent:
1. gather the Influenza Virus material (bud) of FLOS CHRYSANTHEMI ALBA from Haizhou of China improved seeds.
2. FLOS CHRYSANTHEMI ALBA from Haizhou of China bud is cleaned, be placed in flowing water and rinse 10 minutes.
3. carefully peel off outer foil 3-5 and open, take out center bud.
4. pair virulent FLOS CHRYSANTHEMI ALBA from Haizhou of China bud carries out 40-50 ℃ of high temperature, pretreatment in short-term in 5-20 minute.
5. pair FLOS CHRYSANTHEMI ALBA from Haizhou of China bud is successively used 0.1% mercury chloride, 2% clorox sterilization 10 minutes.
6. by the FLOS CHRYSANTHEMI ALBA from Haizhou of China bud of germ-free condition, be placed under stereomicroscope, carry out aseptic dissection, take out the meristematic tissue of 0.2-0.5mm as growing point.
7. under aseptic condition, growing point is placed on optimized cell startup medium and is cultivated 40 days.Obtain virus-free callus.
8. under aseptic condition, virus-free callus is transferred on best FLOS CHRYSANTHEMI ALBA from Haizhou of China propagating culture medium, cultivated and within 30 days, obtain a large amount of Multiple Buds.
9. under aseptic condition, a large amount of Multiple Buds of acquisition are transferred on FLOS CHRYSANTHEMI ALBA from Haizhou of China growth medium, through the cultivation of 30 days, Multiple Buds was grown to serve as unrooted test-tube plantlet.
10. under aseptic condition, unrooted test-tube plantlet is transferred on FLOS CHRYSANTHEMI ALBA from Haizhou of China root media and cultivated 20 days, be grown to serve as the test-tube plantlet that can transplant.
11. under aseptic condition, and virus-free callus is transferred on one-step method medium, cultivates and obtains the test-tube plantlet that has in a large number root in 30 days, provides seedbed to grow seedlings.
12. are transplanted to FLOS CHRYSANTHEMI ALBA from Haizhou of China test-tube plantlet seedling cultivation bed, through the cultivation of the 1-2 month, become detoxification FLOS CHRYSANTHEMI ALBA from Haizhou of China original seed seedling.
13. detoxification FLOS CHRYSANTHEMI ALBA from Haizhou of China original seed seedlings are transplanted in nursery and are carried out the breeding under normal climate condition, can breed then 50-100 detoxification FLOS CHRYSANTHEMI ALBA from Haizhou of China original seed seedlings doubly, provide and produce Demonstration And Extension and breeding.
Claims (1)
1. the cultivation of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds and forming seedling through one step culture quick-breeding method, it is characterized in that, use high temperature to kill the virus of FLOS CHRYSANTHEMI ALBA from Haizhou of China bud, obtain again meristematic tissue and cultivate and transplant, cultivate the test-tube plantlet of virus-free for inducing FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds, producing the FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid a large amount of Multiple Buds of improved seeds test-tube plantlet and the fast numerous FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds test-tube plantlet of forming seedling through one step culture;
In order to reach virus-free effect thoroughly, first virulent FLOS CHRYSANTHEMI ALBA from Haizhou of China bud is carried out to high temperature 40-50 ℃, pretreatment in short-term in 5-20 minute, due to the general non-refractory of virus, therefore adopt high temperature, short time to process, reach part and kill or the effect of passivation virus, can not kill FLOS CHRYSANTHEMI ALBA from Haizhou of China bud again, or reduce the injury to bud simultaneously;
Chrysanthemum Multiple Buds accesses the most effective of induction polyploid in medium after 0.2% colchicine solution is processed 36h, reach 13.3%, by root tip chromosomes Microscopic Identification, the FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds that select high yield and high quality, output further improves 30-50% than dliploid virus-free seedling;
Adopt best FLOS CHRYSANTHEMI ALBA from Haizhou of China propagating culture medium to carry out the induction of a large amount of Multiple Buds of FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds test-tube plantlet, its formula is MS+BA0.2-1.0mg/L+IAA0.05-0.5mg/L, apply this culture medium prescription, monthly the reproduction speed of FLOS CHRYSANTHEMI ALBA from Haizhou of China reaches 20-30 doubly;
Set up one-step method culture technique, FLOS CHRYSANTHEMI ALBA from Haizhou of China polyploid improved seeds test-tube plantlet is only by being once seeded on one-step method medium, and its formula is MS+IAA0.2-1.0mg/L+PP
3330.1-1.0mg/L just can successively complete breeding and two incubation of taking root in 30 days, and application one-step method culture technique, reduces production costs 50%, further accelerates in time reproduction speed simultaneously.
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