CN102499093A

CN102499093A - Novel high-efficient quick virus elimination technique for strawberries

Info

Publication number: CN102499093A
Application number: CN2011103750595A
Authority: CN
Inventors: 高山林
Original assignee: TIANJIN ZHONGZHI BIO-TECH CO LTD
Current assignee: TIANJIN ZHONGZHI BIO-TECH CO LTD
Priority date: 2011-11-23
Filing date: 2011-11-23
Publication date: 2012-06-20

Abstract

The invention relates to a novel high-efficient quick virus elimination technique for strawberries, which comprises four aspects that: firstly a high-temperature short-time inactivation virus technique is adopted to perform short-time pretreatment about 5 to 20 minutes at the high temperature of 40 to 50 DEG C to realize an effect that partial virus is killed or the virus is inactivated, then the size of meristematic tissues for the best virus elimination effect is determined as 0.2 to 0.5mm under a stereoscopic vision microscope through a meristematic tissue size optimization technique, mitosis and growth of virus-free cell clusters are started through a cell starting culture medium formula, a series of optimization tests for the culture medium are carried out through mass Caespitose shoots culture technique to obtain the best strawberry breeding culture medium formula, and achieved annual production of one million of virus-eliminated strawberries seedlings technical level. By adopting the technique, the virus elimination effect can reach 90 to 100 percent, the virus-eliminated strawberry seedlings are applied and experimented on the production for ten years, the quantity of the application trial points reaches more than 100, and the yield increase effect reaches about 50 percent.

Description

The efficient virus-free new technology fast of strawberry

Technical field

The present invention relates to a kind of agro-ecology new technical field, relate in particular to the efficient virus-free new technology fast of a kind of strawberry.

Background technology

The China recent years development is very fast, and, strawberry is easy to infect the several diseases viral disease, has infected the strawberry leaf shrinkage of virus disease, poor growth, and fruit is year by year little, lopsided, poor quality, general underproduction 30-80%.And increase the weight of year by year.The economic benefit of strawberry descends significantly, has seriously hindered the development that strawberry produces.The strawberry of producing simultaneously is owing to poor quality, and fruit is little, lopsided; Therefore the strawberry virus disease is the important disease that influences strawberry output and quality; And at present virus disease effective agricultural chemicals also useless is prevented and treated; Even some agricultural chemicals are arranged can be used; But the virus disease control poor often causes that also residue of pesticide exceed standard, and virus disease has become " cancer " that strawberry produces at present.

For the present only way of strawberry virus disease is exactly to manage to remove the virus in the strawberry seedling body; Be virus-free, some reports are arranged at present, but the difference on effect of removing virus disease be very big both at home and abroad; Technical difficulty is big; Clearance does not wait from 20-60%, but uses the technical method that this research is adopted, and the clearance of virus disease can reach 90-100%.

Summary of the invention

Patent art is set up through the professional research work of carrying out the biotechnology virus-free in more than 20 years; The technology of this project of application, the virus-free effect can reach 90-100%, simultaneously virus-free strawberry seedling application test 10 years on producing; It is individual surplus the application test point reaches 100; The strawberry virus elimination technology has been passed through the appraisal of scientific and technological achievements of Jiangsu Province Science and Technology Department tissue, and the detoxification strawberry that the application patent art cultivates all shows tangible effect of increasing production in the production test of various places, and effect of increasing production reaches about 50%; The general susceptible poison of strawberry is more serious, and the effect of increasing production behind the application virus-free seedling more obviously; Patent art comprises the technology contents of following 4 aspects:

1. high temperature, short time passivation virus technology

In order to reach thorough virus-free effect; At first need carry out high temperature 40-50 ℃ to virulent strawberry bud, preliminary treatment in short-term in 5-20 minute is because the general non-refractory of virus; Therefore we adopt high temperature, short time to handle; Reach the effect that part is killed or passivation is viral, can not kill the strawberry bud again simultaneously, perhaps reduce injury bud.

2. the meristematic tissue optimised is technological

Meristematic tissue is to be positioned at the strawberry bud heart to be in a cell that enlivens splitting status, be generally than less virus, and more near the cell at center, it is just fewer to contain virus, does not perhaps contain virus.Through for many years research repeatedly, confirmed that at last the meristematic tissue size of best virus-free effect is 0.2-0.5mm, cutting growing point must operate under stereomicroscope for this reason.

3. cell starts culture medium prescription

Under stereomicroscope, dissecting the meristematic tissue size that cuts is the key that determines whether to have virus, and meristematic cell group is bigger, and viruliferous possibility is just bigger; Meristematic cell group is littler, and is just malicious in spite of illness, perhaps malicious seldom in spite of illness; Therefore be that cell mass is littler in theory, virus-free is more thorough, and still little cell mass often can not be cultivated and emerge; Difficulty is very big, and for this reason, we are through hundreds of tests; The optimized cell of successfully having set up this patent starts culture medium prescription, and the culture success ratio of the smallest cell group reaches 60%.

4. the bud cultivation of growing thickly in a large number is technological

After the virus-free cell mass of strawberry meristematic tissue is cultivated successfully, also need solve quick breeding, i.e. the problem of cloning; Could application on producing; We have carried out a series of Optimum of culture medium tests, through test for many years in strawberry grows thickly the test of bud medium for this reason; Obtained best strawberry propagating culture medium prescription; Use this culture medium prescription, can reach year a technical merit of producing 1,000,000 strains of virus-free strawberry seedling, can satisfy fully detoxification strawberry large tracts of land produce in to the wilderness demand of strawberry virus elimination test-tube plantlet.

Effect of the present invention: the technology of using this project; The virus-free effect can reach 90-100%, and virus-free strawberry seedling application test 10 years on producing simultaneously is surplus the application test point reaches 100; The strawberry virus elimination technology has been passed through the appraisal of scientific and technological achievements of Jiangsu Province Science and Technology Department tissue; The detoxification strawberry that the application patent art cultivates all shows tangible effect of increasing production in the production test of various places, in general effect of increasing production reaches about 50%; The susceptible poison of strawberry is more serious, and the effect of increasing production behind the application virus-free seedling more obviously.In strawberry virus disease serious area, use the detoxification strawberry seedling that patent art is produced, effect of increasing production can reach 50-100%.

Description of drawings

The strawberry baculoviral electron micrograph that Fig. 1 finds for the present invention;

The granular viral electron micrograph of strawberry that Fig. 2 finds for the present invention;

Fig. 3 is the virus-free effect experiment sketch map of strawberry meristematic tissue size of the present invention.

Embodiment

The techniqueflow of this patent:

1. the susceptible malicious material-bud of strawberrying improved seeds.

2. the strawberry bud is cleaned, placed flowing water flushing 10 minutes.

3. carefully peel off outer foil 3-5 and open, take out the center bud.

4. virulent strawberry bud is carried out high temperature 40-50 ℃ 5-20 minute preliminary treatment in short-term.

5. the strawberry bud is successively used 0.1% mercury chloride, 2% clorox sterilization 10 minutes.

6. with the strawberry bud of germ-free condition, place under the stereomicroscope, carry out aseptic dissection, the meristematic tissue that takes out 0.2-0.5mm is a growing point.

7. under aseptic condition, place optimized cell to start on the medium growing point and cultivated 40 days, and obtained virus-free callus.

8. under aseptic condition, virus-free callus is transferred on the best strawberry propagating culture medium, cultivated and obtained to grow thickly in a large number bud in 30 days.

9. under aseptic condition, the bud that will obtain to grow thickly is in a large number transferred on the strawberry growth medium, and through 30 days cultivation, the blastogenesis of growing thickly grew up to and is the unrooted test-tube plantlet.

10. under aseptic condition, the unrooted test-tube plantlet transferred on the strawberry root media cultivated 20 days, be grown to serve as the test-tube plantlet that can transplant.

11. the strawberry test-tube plantlet is transplanted to seedling cultivation bed,, is become detoxification strawberry original seed seedling through the cultivation of the 1-2 month.Detoxification strawberry original seed seedling is transplanted to the breeding of carrying out in the nursery under the normal climate condition, can breed 50-100 detoxification strawberry original seed seedlings doubly then, provides and produces the demonstration promotion and application.

Claims

1. the efficient virus-free new technology fast of a strawberry, utilization high temperature is killed the part virus of strawberry bud, dissects to cut meristematic tissue and carry out virus-free and cultivate and the cloning quick propagating technology again, cultivates the strawberry test-tube plantlet of virus-free.

2. the efficient virus-free new technology fast of strawberry according to claim 1 is characterized in that at first need carrying out high temperature 40-50 ℃ to virulent strawberry bud in order to reach virus-free effect completely; Preliminary treatment in short-term in 5-20 minute; Because the general non-refractory of virus, so we adopt high temperature, short time to handle, and reach that part is killed or the effect of passivation virus; Simultaneously can not kill the strawberry bud again, perhaps reduce injury bud.

3. the efficient virus-free new technology fast of strawberry according to claim 1 is characterized in that meristematic tissue is to be positioned at the strawberry bud heart to be in a cell that enlivens splitting status, is generally than less virus; And heal near the cell at center; It is just few to contain virus, does not perhaps contain virus, for this reason through for many years research repeatedly; The meristematic tissue size of having confirmed best virus-free effect at last is 0.2-0.5mm, and this growing point must be operated under stereomicroscope.