CN1430881A

CN1430881A - Method for breeding seedlings by using tissue of lauan

Info

Publication number: CN1430881A
Application number: CN 03118063
Authority: CN
Inventors: 彭信海; 谭柏韬; 蒋利媛; 夏晓敏; 罗先权; 陈灵; 刘先芳; 饶红欣
Original assignee: Chenzhou Forestry Science Research Institute; FOREST BOTANICAL GARDEN HUNAN PROV
Current assignee: Chenzhou Forestry Science Research Institute; FOREST BOTANICAL GARDEN HUNAN PROV
Priority date: 2003-01-29
Filing date: 2003-01-29
Publication date: 2003-07-23
Anticipated expiration: 2023-01-29
Also published as: CN1175730C

Abstract

A tissue culture method for culturing the saplings of lauan includes choosing high-quality plant of lauan, taking its auxiliary bud as explant, washing with CaCl2, disinfecting by HgCl2, initial culturing, secondary culturing, rooting culturing for 2 weeks, and culturing in mixed culture medium. Its advantages are high reproduction coefficient (3-4), high rooting rate (95% or mor), and high survival rate (85% or more).

Description

The Liu An group is cultivated seedling-growing method

Technical field: Liu An group is cultivated seedling-growing method and is related to Plant Tissue Breeding, specifically is that the group of a kind of eucalyptus is cultivated seedling-growing method.

Background technology: Liu An (Eucalyptus saligna) is a ma Yao Jin section eucalyptus plant, is the reproducting tree species that a kind of speed is given birth to, material is good.Because Liu An is a cross pollinated species, uses seminal propagation, individual differentiation is big, is difficult to keep the good strains of seeds of Liu An individuality; Simultaneously, the seed seedling is not cold-resistant, and the overwhelming majority freezes to death in the time of-4 ℃; And adopting conventional cottage propagation survival rate lower, nursery stock production cost height does not have using value aborning.Adopting tissue culture method is the main path of exploring the Liu An fast asexual propagation, a few thing has also been done in the group training research of Liu An both at home and abroad, but because some technical problems do not break through, therefore can not have been dropped into large-scale production; The tissue culture method of having studied successful Dunne eucalyptus and cold-resistant eucalyptus camaldulensis also still fails to solve the tissue culture technology problem of Liu An, therefore needs to explore the tissue culture method of Liu An scale nursery stock production.

Summary of the invention: the present invention is intended to study a kind of Liu An group and cultivates seedling-growing method, improves growth coefficient, rooting rate and the transplanting survival rate of Liu An test-tube plantlet, and in conjunction with other relevant technical measures, the large-scale production of Liu An nursery stock is achieved.

The technical scheme that realizes the foregoing invention purpose is: the Liu An tissue culture method comprises explant sterilization, cultivation just, successive transfer culture, culture of rootage and several parts of test-tube seedling transplanting, through HgCl ₂Explant after the sterilization is used CaCl ₂Under aseptic condition, clean, on the MS+BA+IBA medium, carry out just cultivating, test-tube seedling transplanting is cultivated in the mixed-matrix of yellow soil bran adding shell ash, the medium of successive transfer culture and expanding propagation is MS+BA0.1～0.5mg/l+IBA0.5～1.2mg/l+NAA0.1～0.5mg/l, and to be natural scattering light 3000Lux add artificial fill-in light 1500 ± 200Lux with interior to illumination condition; Root media is MS+IBA0.5～1.2mg/l+NAA0.1～0.5mg/l, and illumination condition is that natural scattering light or the following natural scattering light of 5000Lux of 5000～10000Lux adds artificial secondary light source 2500～3000Lux; Through in root media, cultivating about two weeks, move to booth and practice seedling, through about two weeks, move in the mixed-matrix more again and cultivate.

Be described in further detail the present invention below.

The material of Liu An group training will select the axillalry bud of Liu An fine individual plant to make explant, and explant after 30 minutes, is used 0.25%HgCl with flushing with clean water ₂Sterilized 15 minutes, then under aseptic condition with 0.25～1% CaCl ₂Clean 3 times,, use sterile water wash again 3 times to displace the mercury ion that remains on the explant.Explant through sterilization under aseptic condition, is inoculated on the medium of MS+BA0.1～0.5mg/l+IBA0.5～1.2mg/l, at the beginning of under 1000 ± 200Lux illumination condition, cultivate, cultivated about one month, during axillary bud growth to 1～2cm, enter successive transfer culture with expand numerous; Successive transfer culture is MS+BA0.1～0.5mg/l+IBA0.5～1.2mg/l+NAA0.1～0.5mg/l with the medium that expands numerous employing, to be natural scattering light 3000Lux add artificial secondary light source 1500 ± 200Lux with interior to illumination condition, under such condition of culture, 20 days growth coefficient of test-tube plantlet reaches 3～4, for the large-scale production of Liu An nursery stock lays the foundation; Waiting to expand numerous test-tube plantlet breeds after some, the test-tube plantlet of choosing certain altitude carries out culture of rootage, root media is MS+IBA0.5～1.2mg/l+NAA0.1～0.5mg/l, illumination condition is that natural scattering light or the following natural scattering light of 5000Lux of 5000～10000Lux adds artificial secondary light source 2500～3000Lux, under this condition of culture, rooting rate can reach more than 95%; Successive transfer culture and culture of rootage are used part natural scattering light, both can save the energy, and test-tube plantlet is being grown near under the natural environmental condition, improve transplanting survival rate.On the basis of subculture medium that is used for cold-resistant eucalyptus camaldulensis and root media, added NAA, the successive transfer culture of Liu An and the condition of culture of culture of rootage are met, the large-scale production of Liu An is finally realized.At the test-tube plantlet cultivation stage, cultivation temperature is 25 ± 3 ℃, and medium PH is 5.8～6.0, and light application time is 14 hours.Test-tube plantlet is cultivated about a week in root media, begins to take root, and promptly can be moved to booth about two weeks and practices seedling; Practicing seedling is the preceding transition of test-tube seedling transplanting, helps test-tube plantlet progressively to adapt to natural environment, makes nursery stock strong always, and the leaf look deepens, and reaches more than 85% thereby improve the test-tube seedling transplanting survival rate.After the white silk seedling is about 2 weeks, test-tube seedling transplanting is cultivated in the mixed-matrix of yellow soil bran adding shell ash, yellow soil is taken from the following vegetable soil of plow layer, the volume ratio of yellow soil and chaff shell ash is 2: 1, mixed-matrix is not only ventilative, good water permeability, and the essential P of seedling growth, K and other nutritive elements are arranged, also can reduce the harmful microbe invasion and attack.Test-tube seedling transplanting will irrigate with clear water after to the mixed-matrix, and every 3 days spray medicines once, takes place to prevent damage by disease and insect, sprays medicine and is penicillin, fenaminosulf, carbendazim and use in turn for three kinds.The test-tube seedling transplanting place is good with the cement flooring, and is stamped plastic tunnel, and is fine sun-proof, the insulation in winter.Temperature during test-tube seedling transplanting should be controlled between 0～30 ℃, crosses to exceed low the surviving of test-tube plantlet that all be unfavorable for; Under the temperature suitable condition, test-tube seedling transplanting begins growth after 20 days, and later management is identical with general nursery stock.When treating that seedling grows to 8cm, the cuttage expansion that also can carry out tissue cultivating seedling is numerous, and promptly the plugged ear of the semi-lignified about clip 5cm was soaked 30 minutes with 100mg/lIBA, got final product in above-mentioned mixed-matrix or true yellow cubsoil with the conventional method cuttage.

Cultivate the Liu An test-tube plantlet with tissue culture method of the present invention, growth coefficient reached 3～4 in 20 days, and rooting rate reaches more than 95%, and transplanting survival rate reaches more than 85%, and the large-scale production problem of Liu An nursery stock has obtained basic solution; The cuttage of tissue cultivating seedling expansion simultaneously is numerous both can to enlarge the nursery stock production ability, can reduce seedling cost again, had solved the demand of the maximum of nursery stock for large tracts of land popularizing planting Liu An.

Embodiment: following preferred forms for Liu An group cultivation seedling of the present invention.

The axillalry bud of choosing the Liu An fine individual plant is through flushing with clean water, 0.25%HgCl ₂After the sterilization, use 1%CaCl ₂Under aseptic condition, clean 3 times, use sterile water wash again, the first medium of explant is MS+BA0.2mg/l+IBA1.0mg/l, intensity of illumination is 1000Lux, subculture medium is MS+BA0.2mg/l+IBA0.8mg/l+NAA0.2mg/l, to be natural scattering light 3000Lux add artificial fill-in light 1500Lux with interior to illumination condition, the medium of culture of rootage is MS+IBA0.8mg/l+NAA0.2mg/l, illumination condition is that natural scattering light or the following natural scattering light of 5000Lux of 5000～10000Lux adds artificial secondary light source 2500～3000Lux, test-tube plantlet is cultivated about 2 weeks on root media, move into booth and practice seedling, again through moving to yellow soil about 2 weeks: cultivate on the mixed-matrix of old chaff ash=2: 1 (volume ratio), under the temperature suitable condition, transplant and to grow in back 20 days.

Claims

1 ,-and kind of Liu An group cultivation seedling-growing method, comprise explant sterilization, cultivation just, successive transfer culture, culture of rootage and test-tube seedling transplanting, through HgCl ₂Explant after the sterilization is used CaCl ₂Clean under aseptic condition, carry out just cultivating on the MS+BA+IBA medium, test-tube seedling transplanting is cultivated in the mixed-matrix of yellow soil bran adding shell ash, it is characterized in that:

(1) medium of successive transfer culture and expanding propagation is MS+BA0.1～0.5mg/l+IBA0.5～1.2mg/l+NAA0.1～0.5mg/l, and to be natural scattering light 3000Lux add artificial fill-in light 1500 ± 200Lux with interior to illumination condition;

(2) root media is MS+IBA0.5～1.2mg/l+NAA0.1～0.5mg/l, and illumination condition is that natural scattering light or the following natural scattering light of 5000Lux of 5000～10000Lux adds artificial secondary light source 2500～3000Lux;

(3) through in root media, cultivating about two weeks, move to booth and practice seedling, through about two weeks, move in the mixed-matrix more again and cultivate.