CN103190264A - Potato stem tip detoxification method - Google Patents

Abstract

The invention discloses a potato stem tip detoxification method which includes the following steps: A1 sowing pre-basic seed to be detoxified in a field; A2 conducting normal management in the plant growing period; A3 selecting and marking healthy plants from the potato field in the budding period or the flowering period, harvesting the marked plants half a month earlier than other plants and selecting a large-size standard-type healthy potato seed of each plant to be singly harvested and stored; A4 shearing buds to conduct virus detection after the potato seed sprouts; A5 conducting sprouting on the detected healthy potato seeds, shearing large buds which are subjected to alcohol and corrosive sublimate sterilization processing and placing the buds on a common MS solid medium to produce stem seedlings; A6 shearing 1/3 upper parts of the growing stem seedlings to conduct breed conservation and conducting virus detection on the surplus 2/3 parts; and A7 conducting rapid propagation according to a common method after virus detection is qualified. By means of the potato stem tip detoxification method, potato detoxification test tube seedlings can be obtained in a short period, and the obtained detoxification test tube seedlings are healthy and stable in heredity gene.

Description

A kind of potato stem apex poison-removing method

Technical field

The invention provides in the stem apex detoxification of a kind of solution potato and the detoxification process, the stem apex callusization causes genetic mutation and long method of detoxification cycle.This technology belongs to biological technical field.

Background technology

Potato infects virus and causes that degeneration has directly influenced quality and the output of potato in planting process, seriously hindered production and the utilization of potato.Using the potato detoxicating technology, is to solve the major technique measure that seed potato is degenerated.Therefore, virus-free seed potato more and more is subjected to vast potato culture person's welcome.China organizes at the potato stem apex and has also obtained a lot of achievements in research river aspect the cultivation, and builds up the virus-free seed potato breeding of method in various places.Potato stem apex tissue culture detoxicating seedling is the basis of virus-free seed potato industry, and high-quality Shoot Tip Culture rapidly is that time and benefit have been won in the production of later detoxic seedling and virus-free potato.By organizing the cultivation approach, be the present very effective approach that obtains virus-free plant thereby set up virus-free production system.Current toxicity-removing white potato Shoot Tip Culture efficient is low, and planting percent and virus elimination rate do not reach intended purposes, and it is very necessary therefore to optimize potato stem apex detoxification cultural method.

Potato stem apex detoxification standardization program is as follows: stem tuber, plant or budling (perhaps malicious test-tube plantlet) in spite of illness; In conjunction with thermal treatment (36 ℃, illumination in 16 hours; 8 hours dark, 30 ℃); After the surface clean, routine disinfection is handled; Peel off the about 0.1～0.3mm of meristematic tissue under the anatomical lens, be with 1～2 primary leaf original hase; Under aseptic condition, operate; Insert sterilized meristematic tissue medium culture (4-5 month); After treating the meristematic tissue differentiation, change growth medium Cheng Miao (15-20 days) over to; The stem apex differentiation seedling and propagating of tool 5～7 leaves (switching in 15-20 days once); Each strain is that virus and the viroids of stem apex differentiation seedling detects; The test-tube plantlet of no pathogen gives over to basic seedling; At least one season of growth; Industrial fast breeding is used after genetic stability is identified.

This technology is the general procedure of present potato stem apex detoxification, but has many shortcomings:

1, the detoxification cycle is long, link is many, make viral passivation through Overheating Treatment, carries out callus after stem apex is peeled off and becomes seedling to cultivate, and obtaining detoxic seedling also needs to carry out the stability evaluation later on.

2, technology and equipment is required height, need anatomical lens, the operation that stem apex is peeled off is very careful, and stem apex can not be excessive or too small, and excessive virus elimination rate is low, and too small being difficult for survives, and generally requires 0.1-0.3mm, is with 1-2 leaf primordium the most suitable.

3, detoxic seedling may morph, and the callus cultivation meeting after stem apex is peeled off adds some hormones in medium, and meristematic tissue is subjected to these hormonal effects may produce genetic variation, and potato haulm taper born of the same parents are little, and possible genetic material is incomplete.

Summary of the invention

Technical problem to be solved by this invention is to provide a kind of potato stem apex poison-removing method at the deficiencies in the prior art.

Technical scheme of the present invention is as follows:

A kind of potato stem apex poison-removing method may further comprise the steps:

A1, will need the breeder's stock of detoxification to carry out field sowing;

A2, plant strain growth phase Routine Management;

A3, budding or flowering stage the healthy plant of screening in the potato ground, carry out mark, shift to an earlier date the two weeks results than other plant, every strain screening is big, potato type standard and healthy potato seed be results and storing separately;

After A4, potato seed were germinateed, the clip young shoot carried out virus and detects;

A5, will detect healthy potato seed, the big bud of clip is put on the conventional MS solid culture medium and makes stem Duan Miao after alcohol and mercuric chloride sterilization processing after the vernalization again;

A6, with the stem Duan Miao that grows tall, clip top 1/3rd is highly protected kind, residue 2/3rds is carried out virus detection;

A7, virus detect qualified after, carry out numerous soon according to conventional method.

Can in the short relatively cycle, obtain the potato detoxicating cuvette seedling by present technique, the detoxification test tube plantlet health that obtains, gene are stable.Because material is the detoxification breeder's stock, to select through the field forward, the virus elimination rate height has improved detoxification efficient.

Embodiment

Below in conjunction with specific embodiment, the present invention is described in detail.

Potato stem apex poison-removing method of the present invention may further comprise the steps:

1, will need the breeder's stock of detoxification to carry out field sowing.

2, plant strain growth phase Routine Management.

3, budding or flowering stage the healthy plant of screening in the potato ground, carry out mark, shift to an earlier date the two weeks results than other plant, every strain screening is big, potato type standard and healthy potato seed be results and storing separately.

4, after potato seed was germinateed, the clip young shoot carried out virus and detects.

5, will detect healthy potato seed, the big bud of clip is put on the conventional MS solid culture medium and makes stem Duan Miao after alcohol and mercuric chloride sterilization processing after the vernalization again.

6, with the stem Duan Miao that grows tall, clip top 1/3rd is highly protected and is planted, and residue 2/3rds is carried out virus and detected.

7, virus detect qualified after, carry out numerous soon according to conventional method.

Table 1

Annotate: capitalization is represented difference extremely significantly (p＜0.01), lowercase alphabet differential different significantly (p＜0.05).

The technology of the present invention has been simplified the step that stem apex is peeled off, significantly reduced stem apex lift-off technology difficulty, make regeneration plant become strong sprout by weak seedling, become the rapid Cheng Miaowei forming seedling through one step culture of traditional multistep, becoming the seedling time is (to see Table 1) about 20 days, shorten about 33.3%-60% of detoxification cycle, realized the leap in the cycle that shortens from the potato piece to the detoxic seedling long period; Increase planting percent more than 44%, improved the present situation that stem apex is peeled off into the seedling difficulty; Improve virus elimination rate more than 36%, broken through the technical bottleneck that the conventional detoxicity method PVS of potato and PVM are difficult to remove.

The technology of the present invention is started with from shortening detoxification cycle and assurance seed potato genetic stability two links.By with detoxification breeder's stock field sowing, the health control of plant strain growth phase; Squaring period or flowering stage the mark healthy plant; Every strain is gathered in the crops separately, and the screening stature is big, potato type standard and health, treat that it germinates after, its young shoot is carried out virus detection, make stem Duan Miao with detecting qualified potato seed young shoot by aseptic process, stem Duan Miao is carried out virus detect, qualified stem Duan Miao carries out safe preservation as detoxification test tube plantlet, the detoxification test tube plantlet seedling that secures good health, satisfy soilless culture mode needs such as mist training and matrix, thereby reach healthy and potato primary stock production efficiently.

Should be understood that, for those of ordinary skills, can be improved according to the above description or conversion, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.

Claims (1)

1. a potato stem apex poison-removing method is characterized in that, may further comprise the steps:

A1, will need the breeder's stock of detoxification to carry out field sowing;

A2, plant strain growth phase Routine Management;

A3, budding or flowering stage the healthy plant of screening in the potato ground, carry out mark, shift to an earlier date the two weeks results than other plant, every strain screening is big, potato type standard and healthy potato seed be results and storing separately;

After A4, potato seed were germinateed, the clip young shoot carried out virus and detects;

A5, will detect healthy potato seed, the big bud of clip is put on the conventional MS solid culture medium and makes stem Duan Miao after alcohol and mercuric chloride sterilization processing after the vernalization again;

A6, with the stem Duan Miao that grows tall, clip top 1/3rd is highly protected kind, residue 2/3rds is carried out virus detection;

A7, virus detect qualified after, carry out numerous soon according to conventional method.