CN106258370A - Simple Fructus Fragariae Ananssae virus-elimination seedlings mating system - Google Patents

Abstract

The invention belongs to the technical field of horticultural strawberry seedling breeding, and discloses a simple breeding method for virus-free strawberry seedlings. Promote steps such as pumping out stolons of original seedlings (5) sorting and fixing seedlings. The invention reduces the breeding technology of strawberry virus-free seedlings and effectively improves the safety of varieties. The method is simple to operate, easy to master, and highly repeatable, breaking through the conventional laboratory biological breeding technology, reducing labor and material costs, and avoiding the risks brought by open field breeding of diseases and insect pests.

Description

Simple Strawberry Virus-free Seedling Breeding Method

technical field

The invention relates to a simple breeding method for virus-free strawberry seedlings, belonging to the technical field of strawberry seedling breeding.

Background technique

Strawberry is an important berry fruit tree. The fruit is bright in appearance, unique in flavor, high in nutritional value, and widely used. It is known as the "Queen of Fruits". Its cultivated area and output have always ranked first in the production of small berries in the world. According to the statistics of the World Food and Agriculture Organization (FAO), in 2013, the cultivated area of strawberries in the world was 5,424,900 mu, and the output reached 7,739,600 tons; It accounts for 30.40% and 38.73% of the world's total strawberry area and total output, both ranking first in the world.

my country needs more than 160 million detoxified original seedlings every year. According to incomplete statistics, the annual production of detoxified original seedlings in my country is about 15 million, and the market gap for detoxified original seedlings reaches 98%. At present, domestic strawberry virus-free original seedlings mainly use biotechnology for tissue culture and rapid propagation. The technical requirements are difficult to master, difficult to popularize, and the quantity is limited, which is far from meeting market demand. In addition, strawberries reproduce asexually, and the breeding market is chaotic. The original seedlings, virus-free seedlings, common seedlings, and production seedlings are confused. The countermeasure to cultivate robust and high-quality virus-free seedlings and break the bottleneck restricting the development of the strawberry industry is to popularize and promote the breeding technology of virus-free seedlings.

Contents of the invention

The object of the present invention aims at the bottleneck restricting the development of the strawberry industry, and aims to provide a simple strawberry virus-free seedling breeding method.

In order to achieve the above object, the invention provides a kind of simple and easy strawberry virus-free seedling breeding method, its steps comprising:

(1) Acquisition of virus-free original seedlings: cut the newly grown stolon tip from the plant in April, rinse with running water, disinfect, inoculate, and grow for 25 ± 2 days to obtain aseptic seedlings; take the above-mentioned aseptic seedlings for subculture Proliferation, rooting, transplanting to promote survival, and growing for 70±8 days to obtain virus-free original seedlings;

(2) Nursery preparation:

2.1) Site selection and site preparation: The selected site is required to be far away from paddy fields of strawberries, vegetables and solanaceous fruits, and the ground should be leveled. Keep the location and management channel of the original seedlings of virus-free;

2.2) Building a greenhouse: Building a greenhouse on a nursery requires heat preservation in winter, shelter from rain and insects in summer, ventilation and light transmission, and when the light is strong, it can be covered with a sunshade net for protection. precision fertilization;

2.3) 20 days before planting, pour enough water on the substrate and sterilize the shed at high temperature until the ventilation is turned on to cool down before planting;

(3) Changing pots for transplanting and planting: In late July, the original seedlings were detoxified and placed in a greenhouse for hardening, and transplanted after one week of centralized management. Neat, reserve plugs around to breed seedlings, routinely manage fertilizer and water, the greenhouse requires ventilation and light transmission, and when the light is strong, cover it with a sunshade net for protection;

(4) Promote the extraction of stolons of the original seedlings of detoxification: adopt suitable management mode to promote the extraction of stolons of the original seedlings of detoxification;

(5) Arranging and fixing seedlings: After the original seedlings of the virus-free seedlings are pumped out, the top seedlings begin to take root and are arranged and fixed, and the seedlings grow in the plug trays to form independent seedlings.

As a preferred technical solution of the present invention, the method for obtaining the original seedlings of the virus-free method is: in April, the newly grown stolon tip is cut from the plant and rinsed with running water for 1.5-2 hours. Disinfect with 75% medical alcohol for 30 seconds, then disinfect with 0.1% mercury liter water for 8 minutes, and finally rinse with sterile water for 3-4 times, peel off the outer layer, and inoculate the shoot tip on the medium 1 to induce primary culture. Obtain aseptic seedlings in 25±2 days; subculture the above-mentioned aseptic seedlings on culture medium 1 for subculture and multiplication, and transfer the independently grown seedlings to culture medium 2 for rooting culture in 25±2 days; the above operations It is carried out in a culture room at 25±2°C, light intensity of about 2400±200lx, and light time of 14-16 hours per day. Medium one refers to MS+0.5mg/L 6-BA+0.1mg/L NAA+30g/L Sucrose + 6.2g/L agar, pH = 5.8 medium, used to induce primary and secondary proliferation; medium 2 refers to 1/2MS + 30g/L sucrose + 6.2g/L agar, pH = 5.8 medium , for rooting; 25±2 days, remove the virus-free original seedlings that have germinated a complete root system out of the tissue culture room, open the sealing film of the rooted tissue culture seedlings in the room for domestication, place them at room temperature for 1 to 2 days, and remove the root medium Clean it up, transplant it to the volume ratio of peat:perlite:vermiculite 3:2:1, moisturize and promote vitality on the substrate, and obtain the virus-free original seedlings in about 18±2 days.

As a preferred technical solution of the present invention, the edge of the tray filled with the matrix is slightly lower than the surrounding soil, which keeps heat and water; the greenhouse has the functions of heat preservation, rain shelter, insect prevention, ventilation, and high-temperature disinfection of the matrix.

As the preferred technical solution of the present invention, the suitable management mode is: half a month after the potted plants are planted, spray 2‰ water-soluble potassium dihydrogen phosphate and nitrogen fertilizer mixture at intervals of about 10 days to promote pumping, and spray water slightly in the morning and evening Once, the substrate is kept semi-humid, and this mode is managed until mid-October; after November, the temperature drops, and the temperature is not lower than 12°C at night when the shed is closed, and the ventilation is kept at 25-30°C during the day to prevent the plants from growing slowly, and the interval is 15 days Spray 2‰ water-soluble potassium dihydrogen phosphate and nitrogen fertilizer mixture on the left and right sides, and manage in this mode until the seedlings leave the warehouse in mid-March.

As a preferred technical solution of the present invention, the pot diameter used when the original seedlings are repotted and transplanted is 20 cm, and the pot depth is 20 cm.

The technical process of the present invention is simple, easy to grasp, strong in operability, and has the following advantages and effects compared with the prior art knowledge and common means in this field:

The present invention obtains detoxified original seedlings through steps (1) and (2), isolates nurseries, and sterilizes breeding medium substrates to effectively prevent sources and infestation of pests and diseases.

In the present invention, in steps (3) and (4), the detoxified original seedlings proliferate and take root, and after transplanting in late July, no growth regulator is used to ensure the vegetative growth and variety purity of the mother, and prolong the breeding time to promote the increase of the breeding coefficient.

The method provided by the present invention is used in the breeding method of strawberry virus-free seedlings. This method not only breaks through the difficult problem that conventional laboratory biological breeding technology is not easy to popularize, but also prevents the degradation of seed properties and cross-contamination caused by multiple subcultures in conventional laboratories. The safety risk of genetic variation, while reducing the heavy generation work, reducing labor and material costs.

The method provided by the invention is used in the breeding method of virus-free strawberry seedlings, rationally utilizes the functions of greenhouses to prevent insects, avoid rain and keep warm to promote the growth of the mother body and stolon extraction, and avoid the risks brought by plant diseases and insect pests in open field breeding.

The method provided by the present invention is used for the breeding method of strawberry virus-free seedlings. The number of virus-free seedlings can be obtained by the method when it is out of the warehouse in the middle of March, and the breeding coefficient reaches 42-53. It is also very good to connect the production of strawberry seeds before and after Qingming. Seedling planting time.

detailed description

Example 1

(1) Acquisition of the original seedlings of detoxification: On April 20, 2015, the newly grown stolon tip was cut from the plant for washing, disinfection, and inoculation, and the aseptic seedling was obtained on May 14; Multiplication by generation, rooting and transplanting to promote survival, and the original seedlings were obtained on July 23;

(2) Nursery preparation: 1) Choose a paddy field far away from strawberries, vegetables and solanaceous fruits, level the ground, fill the 32-hole tray and tamp the strawberry substrate, and then spread the ground so that the edge of the tray is slightly lower than the surrounding soil, which is convenient for drainage and irrigation. Keep the detoxified original seedlings and management channels; 2) Build a greenhouse on the nursery. The shed film adopts four pieces of polyethylene film, the top is two pieces of crimping, and the other two pieces are skirt films on both sides. Open for ventilation and close to avoid rain. Install insect-proof nets and automatic micro-sprinkler irrigation facilities around; 3) On June 26, the matrix was poured with enough water to sterilize the shed at high temperature, and on July 17, the ventilation was turned on to cool down;

(3) Repotting and transplanting: Put the virus-free original seedlings in the greenhouse on July 27, and transplant them after one week of centralized management. The potted method is adopted. The pot diameter is 20 cm, the pot depth is 20 cm, and the edge of the pot mouth is even with the plug. 48 holes are reserved for breeding seedlings around, the fertilizer and water are routinely managed, the greenhouse is opened for ventilation, and the sunshade net is covered for protection until early September;

(4) Promote detoxification of original seedlings: half a month after planting, spray 2‰ water-soluble potassium dihydrogen phosphate and nitrogen fertilizer mixture at intervals of about 10 days to promote extraction, spray water once in the morning and evening, and keep the substrate half Humid, this mode is managed until mid-October; after November, the temperature drops, and the temperature of the closed shed is not lower than 12°C at night, and the ventilation of the open shed during the day is kept at 25-30°C. ‰Water-soluble potassium dihydrogen phosphate and nitrogen fertilizer mixture, this mode of management until mid-March;

(5) Arranging and fixing seedlings: After the original seedlings of the virus-free seedlings are pumped out, the top seedlings begin to take root and are arranged and fixed, and the seedlings grow in the plug trays to form independent seedlings.

Statistical breeding coefficients of seedlings: the average breeding coefficients of the tested varieties are: Hongjiao 36.68, Zijinjiuhong 53.16. The sources of varieties are: Red Cheek is a Japanese variety introduced in 1999, and Zijin Jiuhong is a new variety independently selected and bred by Jiangsu Academy of Agricultural Sciences, which passed the appraisal of Jiangsu Province in 2015.

Claims (5)

1. a simple Fructus Fragariae Ananssae virus-elimination seedlings mating system, its step includes:

(1) acquisition of detoxification original seed Seedling: the stolon stem apex that April, clip from plant newly grew, flowing water rinse, sterilization, Inoculation, growth obtains aseptic seedling；Take above-mentioned aseptic seedling subculture multiplication, rooting and transplant promotees to live, and growth obtains detoxification original seed Seedling；

(2) nursery prepares:

2.1) addressing, whole: the address challenges of selection is filled also away from Fructus Fragariae Ananssae, the paddy field of vegetable solanaceous vegetables, ground grading, hole tray Tiling earth's surface after compacting substrate, makes hole tray edge be slightly below surrounding soil, and irrigation and drainage are convenient, by reserved detoxification original seed Seedling position and pipe Reason passage；

2.2) booth is built: on nursery, build booth, it is desirable to winter can be incubated, insect protected of taking shelter from rain summer, ventilation and penetrating light, illumination Time strong, sunshade net protection can be added a cover, automatic micro-sprinkling irrigation facility is installed simultaneously, can micro-spray repair water, can precision fertilizing；

2.3) substrate is watered before field planting foot water, and to boil in a covered pot over a slow fire canopy high-temperature sterilization aeration-cooling to field planting front opening；

(3) pot transplanting field planting is changed: late July, by seedling exercising in the original seed rearmounted booth of Seedling detoxification, transplants after 1 week of centralized management, Detoxification original seed Seedling uses potted plant mode, and basin mouth edge is concordant with hole tray, around reserves hole tray and breeds seedling, Routine Management rich water, Booth requires ventilation and penetrating light, when illumination is strong, adds a cover sunshade net protection；

(4) detoxification original seed Seedling stolon pumping is promoted: use suitable management mode to promote detoxification original seed Seedling stolon pumping；

(5) seedling arranges fixing: after detoxification original seed Seedling pumping stolon, and top seedling starts to take root to carry out arranging and fixes, seedling In hole tray, growth forms independent seedling.

Simple Fructus Fragariae Ananssae virus-elimination seedlings mating system the most according to claim 1, it is characterised in that: described detoxification original seed Seedling obtains Access method is: the stolon stem apex that April, clip from plant newly grew rinses 1.5-2h through flowing water, only works aseptic behaviour On platform, first with 75% medical alcohol sterilization 30s, the most again with 0.1% mercuric chloride water sterilization 8min, last aseptic water washing 3-4 time After peel off outer layer bag sheet, stem apex is seeded in culture medium one carry out induce initial culture, 25 ± 2 days obtain aseptic seedling；By upper State aseptic seedling subculture in culture medium one, carry out subculture multiplication expanding propagation, within 25 ± 2 days, the switching of independent seedling will be bred in culture medium Two carry out root culture；Aforesaid operations be 25 ± 2 DEG C, intensity of illumination about 2400 ± 200lx, light application time every day 14～16h Culturing room in carry out, culture medium one refers to MS+0.5mg/L 6-BA+0.1mg/L NAA+30g/L sucrose+6.2g/L agar, The culture medium of pH=5.8, is used for inducing just generation and subculture multiplication；Culture medium two refers to 1/2MS+30g/L sucrose+6.2g/L fine jade Fat, the culture medium of pH=5.8, it is used for taking root；Within 25 ± 2 days, the detoxification original seed Seedling removal group training room of complete root system will be sprouted, in room The sealed membrane inside opening tissue cultured seedling of taking root is tamed, and places 1～2 day, root culture medium cleaned up under normal temperature condition, moves Planting to volume proportion peat: perlite: Vermiculitum is 3: 2: 1, in substrate, moisturizing promotees to live, and within 18 ± 2 days, obtains detoxification original seed Seedling.

Simple Fructus Fragariae Ananssae virus-elimination seedlings mating system the most according to claim 2, it is characterised in that: the cave of described filling substrate Plate edge is slightly below surrounding soil, is incubated water conservation；Booth possesses insulation, takes shelter from rain, insect protected, ventilation function, and substrate is through high-temperature sterilization.

Simple Fructus Fragariae Ananssae virus-elimination seedlings mating system the most according to claim 3, it is characterised in that: described suitable management mould Formula is: after potted plant field planting two weeks, is spaced and within about 10 days, sprays 2 ‰ water-soluble phosphoric acid potassium dihydrogens and nitrogenous fertilizer mixed liquor and promote to take out Raw, once, substrate keeps semi-moist to the most each micro-water spray, and this schema management is to mid-October；Mercury dropped after November, closes canopy Insulation is not less than 12 DEG C night, opens canopy ventilation daytime and keeps 25-30 DEG C, and prevention plant pumping is slow, is spaced and sprays for about 15 days 2 ‰ water-soluble phosphoric acid potassium dihydrogens and nitrogenous fertilizer mixed liquor, this schema management is to seedling outbound mid-March.

Simple Fructus Fragariae Ananssae virus-elimination seedlings mating system the most according to claim 4, it is characterised in that: described original seed Seedling changes basin and moves Basin bore 20cm used, basin deep 20cm during cultivation.