A method for raising eucalyptus regnans

Abstract

The application discloses a seedling raising method of Eucalyptus tereticornis, and selects semi-lignified branches of current-year growth of seedlings as explant materials after Eucalyptus tereticornis seeds are sowed to obtain seedlings, and the seedlings are subjected to disinfection treatment and then are subjected to induction culture, subculture and rooting culture, and finally, the rooting bottle seedlings with complete rooting and 3-4cm seedling height are transplanted. The seedlings are obtained through sowing, and the early growth excellent Eucalyptus tereticornis seedlings are selected as the raw materials of the tissue culture explants. The mature leaves of Eucalyptus tereticornis have high leatherization degree, and the difficulty of obtaining latent buds increases with the increase of age. The 1-year-old Eucalyptus tereticornis explants are easier to induce sterile buds. After induction, subculture and rooting culture, the rooting rate is more than 90%, and the transplanting survival rate is 90%.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of plant cultivation, and relates to a seedling raising method of Corymbia flavescens. BACKGROUND

[0002] Corymbia flavescens is a tree species of Myrtaceae, with a height of 15 meters, forming a woody tuber. The bark is smooth, powdery, and falls off in thin scales of ash yellow, showing bright white. It is originally from Australia (Northern Territory, Queensland, Western Australia), and has not been introduced in China. In recent years, the development of eucalyptus artificial industrial raw material forest in China is very rapid, and the planting scale is continuously expanding. Corymbia flavescens has certain cold resistance, and has broad prospects for popularization in cold regions.

[0003] The tissue culture systems of different strains of eucalyptus are quite different, and the tissue culture system of different single plants, especially excellent single plants, basically needs to be re-established. Therefore, in order to better and faster popularize Corymbia flavescens, it is necessary to break through the tissue culture and rapid propagation technology according to the physiological and biochemical characteristics of Corymbia flavescens. SUMMARY

[0004] The present application provides a seedling raising method of Corymbia flavescens, which realizes tissue culture and rapid propagation of Corymbia flavescens, and can improve the rooting rate and transplanting survival rate of tissue culture.

[0005] The technical scheme adopted by the present application is as follows:

[0006] A seedling raising method of Corymbia flavescens obtains seedlings by sowing Corymbia flavescens seeds, selects semi-lignified branches of current-year seedlings as explant materials, performs induction culture, subculture and rooting culture after disinfection, and finally transplants the rooting bottle seedlings with complete rooting and a seedling height of 3-4 cm;

[0007] The main steps include:

[0008] (1) Sowing and seedling raising

[0009] Mix Corymbia flavescens seeds and sowing substrate uniformly according to a weight ratio of 1:0.5, spread on the sowing substrate, then cover a layer of sowing substrate, take the seedlings as the degree, spray with a sprayer, keep the sowing substrate moist, and cover a film.

[0010] (2) Transplanting of seedlings

[0011] When the Corymbia flavescens after sowing grows 3 pairs of leaves and the seedling height is 4-5 cm, transplanting is performed, a small hole is punched on a seedling raising cup containing transplanting substrate with a small stick, then the Corymbia flavescens seedlings are transplanted, the seedlings are sprayed with rooting water, the cup is covered with a film after arching, and a shading net is finally covered; after 120-150 days of transplanting, the height of the Corymbia flavescens seedling is 30-35 cm, and the ground diameter is 3-3.5 mm, and the seedling is out of the nursery.

[0012] (3) Disinfection of explants

[0013] Semi-lignified branches of the current-year Eucalyptus leucoxylon seedlings were selected after the nursery, and 2-3 cm stem segments were cut as explants. The explants were immersed in 75% alcohol for 20 s, washed with sterile water for 2-3 times, then immersed in 2% sodium hypochlorite for 2 min, and finally washed with sterile water for 2-3 times. The explants were dried with filter paper and prepared for use;

[0014] (4) Induction culture

[0015] The explants treated in step (3) were inoculated on the induction medium for induction culture. After dark culture at 25±2℃ for 5-7 d, the culture was carried out under the conditions of 12 h / d illumination and 1500-2000 lux illumination intensity for 20-25 d;

[0016] (5) Subculture

[0017] The aseptic sprouts obtained in step (4) were cut and transferred to the subculture medium. After dark culture at 25±2℃ for 5-7 d, the culture was carried out under the conditions of 12 h / d illumination and 2000-2500 lux illumination intensity for 20-25 d;

[0018] (6) Rooting culture

[0019] When the sprouts obtained in step (5) grew to 3-5 cm, the sprouts were cut and transferred to the rooting medium. After dark culture at 25±2℃ for 5-7 d, the culture was carried out under the conditions of 12 h / d illumination and 2000-2500 lux illumination intensity for 20-25 d;

[0020] (7) Transplanting of tissue culture seedlings

[0021] The rooting bottle seedlings with complete roots and 3-4 cm in height were washed of the culture medium and transplanted into nutrient cups containing transplanting substrate. The transplanting substrate was soaked, covered with a moisture-retaining film and a shading net, and sprayed with a 500-600-fold solution of carbendazim every 5-7 d to prevent root rot. After 10 d, the shading net and the film were removed, and leaf nutrient solution was sprayed every 10 d.

[0022] Preferably, the raw material components of the induction medium in step (4) above are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / L Ca(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.3-0.5 mg / L 6-BA, 0.1-0.2 mg / L NAA, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.2.

[0023] Preferably, the raw material components of the subculture medium in step (5) above are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / L Ca(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.3-0.5 mg / L 6-BA, 0.2-0.4 mg / L NAA, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.2.

[0024] Preferably, the rooting medium raw material components of the above step (6) are: 900 mg / L KNO3, 180 mg / L NH4NO3, 135 mg / L KH2PO4, 100 mg / L CaCl2, 89 mg / L Ca(NO3)2, 180 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.4-0.6 mg / L IBA, 0.3-0.5 mg / L ABT1, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.2.

[0025] Preferably, the seeding substrate of the above step (1) is a mixture of yellow heart soil and clean river sand in a weight ratio of 1:1, which is sterilized with a 0.5% volume concentration potassium permanganate solution, covered with a film and sealed for one day, then uncovered and dried, and then soaked with water for use.

[0026] Preferably, the seedling is sprayed with thiophanate-methyl or chlorothalonil every 7 days after the above step (2) to prevent diseases, and then sprayed with a 0.2-0.5% volume concentration compound fertilizer solution every 10 days after the new roots of the Eucalyptus leucophloia seedlings grow.

[0027] Preferably, the transplanting substrate of the above steps (2) and (7) is a mixture of coconut husk, carbonized rice husk, peat soil and yellow heart soil in a weight ratio of 6:2:1:1, to which 4 kg / m3 of calcium-magnesium-phosphorus fertilizer is added. 3 , which is obtained after being covered with a film and composted under the sun for one month to fully ferment.

[0028] Preferably, the raw material components of the nutrient solution of the above step (7) are: 1 L of the foliar nutrient solution contains 180 mg KNO3, 36 mg NH4NO3, 27 mg KH2PO4, 20 mg CaCl2, 36 mg MgSO4, 1 mg H3BO3, 2.5 mg MnSO4, 1 mg ZnSO4, 0.025 mg Na2MoO4, 0.0025 mg CuSO4, 0.0025 mg CoCl2, and 0.083 mg KI.

[0029] The application has the following advantages and beneficial effects:

[0030] 1. The application firstly obtains seedling seedlings by sowing, and selects early growth excellent Eucalyptus citriodora seedling seedlings as the raw material of tissue culture explants, because the mature leaf of Eucalyptus citriodora has high degree of leatherization, and the difficulty of latent bud acquisition increases with age, so that it is easier to induce sterile buds by selecting 1-year-old Eucalyptus citriodora explants.

[0031] 2. On the basis of induction culture and subculture, the rooting medium of the application reduces the use of a large amount of elements and part of trace elements, and through the collocation between different elements, the rooting rate and quality of Eucalyptus citriodora are ensured, and finally the in-vitro rooting rate can reach more than 90%, which lays a foundation for transplanting.

[0032] 3. The application selects specific substrates when transplanting, which ensures the comprehensive and sufficient nutrition required for the growth and survival of Eucalyptus citriodora, effectively improves the transplanting survival rate of Eucalyptus citriodora tissue culture seedlings, and the transplanting survival rate can reach more than 90%, reduces material loss, and promotes the large-scale production of Eucalyptus citriodora.

[0033] 4. There are many Eucalyptus hybrid line tissue culture systems, but the differences between different Eucalyptus lines are still relatively large, and the tissue culture system of different single plants, especially excellent single plants, basically needs to be re-established. The application establishes the tissue culture system of Eucalyptus citriodora excellent single plants, promotes the development of Eucalyptus citriodora tissue culture seedling factory production, is conducive to the popularization and planting of Eucalyptus citriodora varieties, and has good economic and ecological benefits. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 It is a seedling seedling figure after sowing and raising by the application.

[0035] Figure 2 It is a seedling seedling figure after sowing and raising by the application.

[0036] Figure 3 It is a subculture seedling figure obtained by subculture according to the application.

[0037] Figure 4 It is a rooting bottle seedling figure obtained by rooting culture according to the application.

[0038] Figure 5 It is a scene after transplanting management of the tissue culture seedling of the application Figure I .

[0039] Figure 6 It is a scene after transplanting management of the tissue culture seedling of the application Figure II . DETAILED DESCRIPTION

[0040] The application will be further described below in combination with examples. Embodiment

[0041] A eucalyptus regnans seedling raising method,

[0042] The main steps include:

[0043] (1) Sowing seedling raising

[0044] Mix the eucalyptus regnans seeds and the sowing substrate uniformly according to the weight ratio of 1:0.5, spread on the sowing substrate, then cover a layer of sowing substrate, and cover the film after spraying water with a sprayer to keep the sowing substrate moist;

[0045] The sowing substrate is mixed by the yellow heart soil and clean river sand according to the weight ratio of 1:1, disinfected by 0.5% potassium permanganate solution, then covered with a film, sealed for one day, uncovered and dried after the film is uncovered, and then soaked with water for standby;

[0046] (2) Transplanting seedling

[0047] When the eucalyptus regnans grows 3 pairs of leaves and the seedling height is 4-5 cm after sowing, a small hole is inserted on the seedling cup filled with the transplanting substrate by a small stick, then the eucalyptus regnans seedling is transplanted, soaked with the rooting water, covered with a film after the arch is raised, and then covered with a shading net;

[0048] The transplanting substrate is mixed by the coconut husk, carbonized rice husk, peat soil and yellow heart soil according to the weight ratio of 6:2:1:1, then 4 kg / m 3 , and obtained after covering with a film under the sun for one month for fermentation;

[0049] The seedling is transplanted every 7 days, and the seedling is sprayed with the methyldithane according to the conventional method to prevent diseases, the seedling is sprayed with the compound fertilizer water solution with the volume concentration of 0.2-0.5% every 10 days when the eucalyptus regnans seedling grows new roots, and the eucalyptus regnans seedling is transplanted out of the nursery after 120-150 days after the seedling is transplanted, the height of the eucalyptus regnans seedling is 30-35 cm, and the ground diameter is 3-3.5 mm;

[0050] (3) Disinfection of explants

[0051] The semi-lignified branches of the current-year eucalyptus regnans seedling transplanted out of the nursery are selected, 2-3 cm stem segments are cut as explants, the explants are soaked with 75% alcohol for 20 s, washed with sterile water for 2-3 times, then soaked with 2% sodium hypochlorite for 2 min, finally washed with sterile water for 2-3 times, dried with filter paper, and standby;

[0052] (4) Induction culture

[0053] The explant treated in step (3) is inoculated on the induction medium for induction culture. After dark culture at 25±2℃ for 5-7 days, the explant is cultured under the condition of 12h / d illumination and 1500-2000 lux illumination intensity for 20-25 days;

[0054] The raw material components of the induction medium are: 1800mg / L KNO3, 360mg / L NH4NO3, 270mg / L KH2PO4, 200mg / L CaCl2, 178mg / L Ca(NO3)2, 360mg / L MgSO4, 10mg / L H3BO3, 25mg / L MnSO4, 10mg / L ZnSO4, 0.25mg / L Na2MoO4, 0.025mg / L CuSO4, 0.025mg / L CoCl2, 0.83mg / L KI, 1000mg / L PVP, 2.4mg / L D-calcium pantothenate (VB5), 0.24mg / L biotin (VH), 100mg / L L-cysteine, 2mg / L glycine, 5mg / L nicotinic acid (VB3), 2mg / L thiamine (VB1), 0.5mg / L pyridoxine (VB6), 37.3mg / L Na2-EDTA, 27.8mg / L FeSO4, 100mg / L myo-inositol, 0.3mg / L 6-BA, 0.2mg / L NAA, 30g / L sucrose, 8g / L agar powder, and the pH value is 5.8-6.2;

[0055] (5) Subculture

[0056] The sterile bud obtained in step (4) is transferred into the subculture medium, and after dark culture at 25±2℃ for 5-7 days, the explant is cultured under the condition of 12h / d illumination and 2000-2500 lux illumination intensity for 20-25 days;

[0057] The raw material components of the subculture medium are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / L Ca(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.3 mg / L 6-BA, 0.4 mg / L NAA, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.0;

[0058] (6) Rooting culture

[0059] When the bud seedlings obtained in step (5) grow to 3-5 cm, the bud seedlings are cut and transferred into a rooting medium, and after dark culture at 25±2℃ for 5-7 days, the culture is carried out under the conditions of light illumination of 12 h / d and light intensity of 2000-2500 lux for 20-25 days;

[0060] The rooting medium raw material components are: 900 mg / L KNO3, 180 mg / L NH4NO3, 135 mg / L KH2PO4, 100 mg / L CaCl2, 89 mg / L Ca(NO3)2, 180 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.4 mg / L IBA, 0.3 mg / L ABT1, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.0;

[0061] (7) Transplanting of the tissue culture seedlings

[0062] The rooting complete and 3-4 cm high seedlings are washed with the medium and then transplanted into a nutrient cup containing the transplanting substrate, the transplanting substrate is soaked, a film is covered for moisture retention, a shading net is covered, and a 500-600 times solution of carbendazim is sprayed every 5-7 days to prevent root rot; the shading net and the film are opened after 10 days, and a foliar nutrient solution is sprayed every 10 days;

[0063] The foliar nutrient solution raw material components are: 1 L of the foliar nutrient solution contains 180 mg of KNO3, 36 mg of NH4NO3, 27 mg of KH2PO4, 20 mg of CaCl2, 36 mg of MgSO4, 1 mg of H3BO3, 2.5 mg of MnSO4, 1 mg of ZnSO4, 0.025 mg of Na2MoO4, 0.0025 mg of CuSO4, 0.0025 mg of CoCl2, and 0.083 mg of KI.

[0064] The transplanting substrate is obtained by uniformly mixing coconut husk, carbonized rice husk, peat soil and yellow heart soil at a weight ratio of 6:2:1:1, and then adding 4 kg / m3 of calcium-magnesium-phosphorus fertilizer. 3 The coconut husk, the carbonized rice husk, the peat soil and the yellow heart soil are mixed and fermented under the sun for one month. Embodiment

[0065] A yellow leaf eucalyptus seedling raising method mainly includes the following steps:

[0066] (1) Seedling raising

[0067] Mix the seed of yellow leaf eucalyptus and the seeding substrate uniformly according to the weight ratio of 1:0.5, spread on the seeding substrate, then cover a layer of seeding substrate, so as not to see the seed, spray with a sprayer, keep the seeding substrate moist, and cover the film;

[0068] The seeding substrate is mixed by yellow heart soil and clean river sand according to the weight ratio of 1:1, disinfected by 0.5% potassium permanganate solution, then covered with a film, sealed for one day, uncovered the film after airing, and then soaked with water for standby;

[0069] (2) Transplanting of seedling

[0070] When the yellow leaf eucalyptus grows 3 pairs of leaves and the height of seedling is 4-5 cm after seeding, a small hole is inserted on the seedling cup filled with transplanting substrate by a small stick, then the yellow leaf eucalyptus seedling is transplanted into the hole, soaked with rooting water, covered with a film after arching to keep moist, and finally covered with shading net;

[0071] The transplanting substrate is mixed by coconut husk, carbonized rice husk, peat soil and yellow heart soil according to the weight ratio of 6:2:1:1, then 4 kg / m 3 , and then covered with a film under the sun for one month to ferment;

[0072] After the seedling is transplanted, methylthiophanate is sprayed every 7 days according to the conventional method to prevent diseases, and when the yellow leaf eucalyptus seedling grows new roots, compound fertilizer solution with a volume concentration of 0.2-0.5% is sprayed every 10 days; After 120-150 days after transplanting, the height of the yellow leaf eucalyptus seedling is 30-35 cm, and the ground diameter is 3-3.5 mm, and the seedling is out of the nursery;

[0073] (3) Disinfection of explants

[0074] The semi-lignified branches of the current year's yellow leaf eucalyptus seedling are selected after the seedling is out of the nursery, and 2-3 cm stem segments are cut as explants. The explants are soaked in 75% alcohol for 20 s, washed with sterile water for 2-3 times, then soaked in 2% sodium hypochlorite for 2 min, finally washed with sterile water for 2-3 times, dried with filter paper, and standby;

[0075] (4) Induction culture

[0076] The treated explants in step (3) are inoculated on the induction medium for induction culture, and cultured in the dark for 5-7 days at a temperature of 25±2℃, and then cultured under the conditions of light for 12 h / d and light intensity of 1500-2000 lux for 20-25 days;

[0077] The raw material components of the induction medium are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / L Ca(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.4 mg / L 6-BA, 0.2 mg / L NAA, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.0;

[0078] (5) Subculture

[0079] The sterile buds obtained in step (4) are transferred into a subculture medium, and after dark culture at a temperature of 25±2°C for 5-7 days, the culture is carried out under the conditions of light illumination for 12 h / d and light intensity of 2000-2500 lux for 20-25 days;

[0080] The raw material components of the subculture medium are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / L Ca(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.4 mg / L 6-BA, 0.3 mg / L NAA, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.0.

[0081] (6) Rooting culture

[0082] When the shoots obtained in step (5) grow to 3-5 cm, the shoots are cut and transferred to a rooting medium, and after dark culture at 25±2℃ for 5-7 days, the shoots are cultured under light of 12h / d and light intensity of 2000-2500 lux for 20-25 days;

[0083] The raw material components of the rooting medium are: 900 mg / L KNO3, 180 mg / L NH4NO3, 135 mg / L KH2PO4, 100 mg / L CaCl2, 89 mg / L Ca(NO3)2, 180 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L calcium D-pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.5 mg / L IBA, 0.4 mg / L ABT1, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 5.8-6.0;

[0084] (7) Transplanting of the tissue culture seedlings

[0085] The rooting seedlings with complete roots and a height of 3-4 cm are washed of the culture medium and transplanted into nutrient cups containing a transplanting substrate, the transplanting substrate is soaked, a film is covered for moisture retention and a shading net is covered, and a 500-600 times solution of carbendazim is sprayed every 5-7 days to prevent root rot; after 10 days, the shading net and the film are opened, and a foliar nutrient solution is sprayed every 10 days;

[0086] The raw material components of the foliar nutrient solution are: 1L of the foliar nutrient solution contains 180 mg KNO3, 36 mg NH4NO3, 27 mg KH2PO4, 20 mg CaCl2, 36 mg MgSO4, 1 mg H3BO3, 2.5 mg MnSO4, 1 mg ZnSO4, 0.025 mg Na2MoO4, 0.0025 mg CuSO4, 0.0025 mg CoCl2, and 0.083 mg KI;

[0087] The transplanting medium is made by mixing coconut husk, carbonized rice husk, peat soil and loess soil in a weight ratio of 6:2:1:1, and then adding 4kg / m2 of calcium magnesium phosphate fertilizer. 3 , covered with a film and piled up in the sun for a month to allow it to fully ferment. Example

[0088] A method for raising seedlings of Eucalyptus erythrorhizome, the main steps of which include:

[0089] (1) Sowing and seedling raising

[0090] Mix the Eucalyptus eryngii seeds and the sowing substrate in a weight ratio of 1:0.5, sow on the sowing substrate, then cover with a layer of sowing substrate until the seeds are no longer visible, water thoroughly with a sprayer, and cover with a film to keep the sowing substrate moist;

[0091] The sowing medium is a mixture of loess soil and clean river sand in a weight ratio of 1:1, disinfected with a 0.5% volume concentration of potassium permanganate solution, then covered with a film and sealed for one day. After uncovering the film and drying in the sun, soak it with water and set aside.

[0092] (2) Transplanting seedlings

[0093] After sowing, the yellow leaf eucalyptus grows 3 pairs of leaves and the seedling height is 4-5cm. Transplant it. Use a small wooden stick to poke a small hole in the seedling cup filled with transplanting medium, then plant the yellow leaf eucalyptus seedling, drench it with water to establish roots, cover it with film to keep it moist after it arches, and finally cover it with a shade net.

[0094] The transplanting medium is made by mixing coconut husk, carbonized rice husk, peat soil and loess soil in a weight ratio of 6:2:1:1, and then adding 4kg / m2 of calcium magnesium phosphate fertilizer. 3 , covered with film and piled up in the sun for one month to allow it to fully ferment;

[0095] After transplanting, spray the seedlings with chlorothalonil every 7 days to prevent diseases. When the yellow-leaf eucalyptus seedlings grow new roots, spray them with a 0.2-0.5% compound fertilizer solution every 10 days. 120-150 days after transplanting, the yellow-leaf eucalyptus seedlings should be transplanted when they are 30-35 cm tall and 3-3.5 mm in diameter at ground level.

[0096] (3) Explant disinfection

[0097] Select semi-lignified branches of Eucalyptus chrysantha seedlings of the current year after transplanting, cut 2-3 cm stem segments as explants, soak the explants in 75% alcohol for 20 seconds, wash them 2-3 times with sterile water, then soak them in 2% sodium hypochlorite for 2 minutes, finally wash them 2-3 times with sterile water, blot them dry with filter paper, and set aside;

[0098] (4) Induction culture

[0099] The explant treated in step (3) is inoculated on the induction medium for induction culture, and after dark culture at 25±2℃ for 5-7 days, it is cultured under the condition of 12h / d illumination and 1500-2000 lux illumination intensity for 20-25 days;

[0100] The raw material components of the induction medium are: 1800mg / L KNO3, 360mg / L NH4NO3, 270mg / L KH2PO4, 200mg / L CaCl2, 178mg / L Ca(NO3)2, 360mg / L MgSO4, 10mg / L H3BO3, 25mg / L MnSO4, 10mg / L ZnSO4, 0.25mg / L Na2MoO4, 0.025mg / L CuSO4, 0.025mg / L CoCl2, 0.83mg / L KI, 1000mg / L PVP, 2.4mg / L D-calcium pantothenate (VB5), 0.24mg / L biotin (VH), 100mg / L L-cysteine, 2mg / L glycine, 5mg / L nicotinic acid (VB3), 2mg / L thiamine (VB1), 0.5mg / L pyridoxine (VB6), 37.3mg / L Na2-EDTA, 27.8mg / L FeSO4, 100mg / L myo-inositol, 0.5mg / L 6-BA, 0.1mg / L NAA, 30g / L sucrose, 8g / L agar powder, and the pH value is 6.0-6.2;

[0101] (5) Subculture

[0102] The sterile bud obtained in step (4) is transferred to the subculture medium, and after dark culture at 25±2℃ for 5-7 days, it is cultured under the condition of 12h / d illumination and 2000-2500 lux illumination intensity for 20-25 days;

[0103] The raw material components of the subculture medium are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / L Ca(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.5 mg / L 6-BA, 0.2 mg / L NAA, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 6.0-6.2;

[0104] (6) Rooting culture

[0105] When the bud seedlings obtained in step (5) grow to 3-5 cm, the bud seedlings are cut and transferred into a rooting medium, and after dark culture at 25±2°C for 5-7 days, the culture is carried out under the conditions of light illumination of 12 h / d and light intensity of 2000-2500 lux for 20-25 days;

[0106] The rooting medium raw material components are: 900 mg / L KNO3, 180 mg / L NH4NO3, 135 mg / L KH2PO4, 100 mg / L CaCl2, 89 mg / L Ca(NO3)2, 180 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / L MnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate (VB5), 0.24 mg / L biotin (VH), 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid (VB3), 2 mg / L thiamine (VB1), 0.5 mg / L pyridoxine (VB6), 37.3 mg / L Na2-EDTA, 27.8 mg / L FeSO4, 100 mg / L myo-inositol, 0.6 mg / L IBA, 0.3 mg / L ABT1, 30 g / L sucrose, 8 g / L agar powder, and the pH value is 6.0-6.2;

[0107] (7) Transplanting of the tissue culture seedlings

[0108] The rooting bottle seedlings with complete rooting and 3-4 cm in height are washed with the culture medium, transplanted into the nutrient cups containing the transplanting substrate, the transplanting substrate is soaked, covered with a moisture-retaining film and a shading net, and sprayed with a 500-600 times solution of carbendazim every 5-7 days to prevent root rot; the shading net and the film are opened after 10 days, and the leaf nutrient solution is sprayed every 10 days;

[0109] The raw material components of the leaf nutrient solution are: 1 L of the leaf nutrient solution contains 180 mg of KNO3, 36 mg of NH4NO3, 27 mg of KH2PO4, 20 mg of CaCl2, 36 mg of MgSO4, 1 mg of H3BO3, 2.5 mg of MnSO4, 1 mg of ZnSO4, 0.025 mg of Na2MoO4, 0.0025 mg of CuSO4, 0.0025 mg of CoCl2, and 0.083 mg of KI.

[0110] The transplanting substrate is obtained by uniformly mixing coconut husk, carbonized rice husk, peat soil and yellow heart soil at a weight ratio of 6:2:1:1, and then adding 4 kg / m3 of calcium-magnesium-phosphorus fertilizer. 3 The mixture is fermented for one month under the sun with a film covering.

[0111] Comparative Example 1

[0112] The difference between the example 3 is only that, without the step (1) and step (2) of example 3, step (3) selects the excellent single plant of 2-year-old Eucalyptus maculata real seedlings in Eucalyptus maculata forest as the mother tree of explant raw material, takes the semi-lignified branches of this year, cuts 2-3cm stem segments as explants, soaks the explants with 75% alcohol for 20s, washes with sterile water for 2-3 times, then soaks with 2% sodium hypochlorite for 2min, finally washes with sterile water for 2-3 times, absorbs with filter paper, and stands by.

[0113] Comparative example 2:

[0114] The difference between the example 3 is only that, step (6) is different in the raw material components of rooting medium, the use amount of K, N, P, Ca, Mg and other elements is increased, specifically, the rooting medium raw material components used in this comparative example are: 1800mg / L KNO3, 360mg / L NH4NO3, 270mg / L KH2PO4, 200mg / L CaCl2, 178mg / L Ca(NO3)2, 360mg / L MgSO4, 10mg / L H3BO3, 25mg / L MnSO4, 10mg / L ZnSO4, 0.25mg / L Na2MoO4, 0.025mg / L CuSO4, 0.025mg / L CoCl2, 0.83mg / L KI, 1000mg / L PVP, 2.4mg / L calcium D-pantothenate (VB5), 0.24mg / L biotin (VH), 100mg / L L-cysteine, 2mg / L glycine, 5mg / L nicotinic acid (VB3), 2mg / L thiamine (VB1), 0.5mg / L pyridoxine (VB6), 37.3mg / L Na2-EDTA, 27.8mg / L FeSO4, 100mg / L myo-inositol, 0.6mg / L IBA, 0.3mg / L ABT1, 30g / L sucrose, 8g / L agar powder, and the pH value is 6.0-6.2.

[0115] Comparative example 3:

[0116] The difference between the example 3 is only that, the raw material components of foliar nutrient solution in step (7) are different, the foliar nutrient solution of this comparative example is a nitrogen, phosphorus and potassium mixed solution (1L mixed solution contains 20-30g superphosphate, 3-5g urea and 10-15g potassium sulfate).

[0117] Test example:

[0118] The in-bottle rooting rate and average root length of Eucalyptus maculata tissue culture seedlings of examples 1-3 and comparative examples 1-2, and the survival rate after three months of transplanting are calculated, and the results are as follows:

[0119] Group In-bottle rooting rate Average root length of bottle seedling Transplanting survival rate Example 1 93.5% 2.9 92.8% Example 2 95.2% 2.9 95.0% Example 3 91.8% 2.8 92.1% Comparative Example 1 78.9% 2.6 80.2% Comparative Example 2 71.7% 2.3 69.8% Comparative Example 3 91.2% 2.8 81.5%

[0120] From the above table, the technology of the present application is a complete technical solution, especially after the complete steps of seeding and seedling, transplanting of seedling, rooting culture and post-transplanting nutrient solution spraying of the present application, the rooting rate of Eucalyptus tereticornis tissue culture seedlings in the bottle can be increased to more than 90%, and the transplanting survival rate can be increased to more than 90%.

Claims

1. A method for raising seedlings of Eucalyptus erythrorhizome, characterized in that: After sowing Eucalyptus fulva seeds to obtain seedlings, semi-lignified branches of the seedlings grown in the current year were selected as explant materials, and after disinfection, they were used for induction culture, subculture and rooting culture. Finally, the rooted seedlings with complete rooting and a height of 3-4 cm were transplanted; The main steps include: (1) Sowing and seedling raising Mix the Eucalyptus eryngii seeds and the sowing substrate in a weight ratio of 1:0.5, sow on the sowing substrate, then cover with a layer of sowing substrate until the seeds are no longer visible, water thoroughly with a sprayer, and cover with a film to keep the sowing substrate moist; (2) Transplanting seedlings After sowing, the yellow-leaf eucalyptus grows 3 pairs of leaves and the seedlings are 4-5 cm tall before transplanting. Use a small wooden stick to poke a small hole in the seedling cup filled with transplanting medium, then plant the yellow-leaf eucalyptus seedlings, drench them with water to establish roots, cover them with film to keep them moist after arching, and finally cover them with shade nets. 120-150 days after transplanting, the yellow-leaf eucalyptus seedlings are transplanted when they are 30-35 cm tall and 3-3.5 mm in diameter at ground level. (3) Explant disinfection Select semi-lignified branches of Eucalyptus chrysantha seedlings of the current year after transplanting, cut 2-3 cm stem segments as explants, soak the explants in 75% alcohol for 20 seconds, wash them 2-3 times with sterile water, then soak them in 2% sodium hypochlorite for 2 minutes, finally wash them 2-3 times with sterile water, blot them dry with filter paper, and set aside; (4) Induction culture The explants treated in step (3) were inoculated onto an induction medium for induction culture. After culturing in the dark at a temperature of 25±2°C for 5-7 days, they were then cultured under the conditions of 12 h / d of light and 1500-2000 lux for 20-25 days. (5) Subculture The sterile buds obtained in step (4) were cut and transferred to subculture medium. After culturing in the dark at a temperature of 25±2°C for 5-7 days, they were cultured under the conditions of 12 hours / day of light and 2000-2500 lux of light intensity for 20-25 days. (6) Rooting culture When the seedlings obtained in step (5) grow to 3-5 cm, cut the seedlings and transfer them to rooting medium. After culturing in the dark at 25±2°C for 5-7 days, they are cultured under the conditions of 12 h / d of light and 2000-2500 lux for 20-25 days. (7) Transplanting tissue culture seedlings After the rooted seedlings have fully rooted and are 3-4 cm tall, wash the culture medium and transplant them into a nutrient cup filled with transplanting medium. Drench the transplanting medium thoroughly, cover with a film to keep it moist, and cover with a shade net. Spray with 500-600 times diluted carbendazim every 5-7 days to prevent root rot. After 10 days, open the shade net and film, and spray the leaves with nutrient solution every 10 days. The raw material components of the induction culture medium in step (4) are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / LCa(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / LMnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-pantothenate calcium, 0.24 mg / L biotin, 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid, 2 mg / L thiamine, 0.5 mg / L pyridoxine, 37.3 mg / L Na2-EDTA, 27.8mg / L FeSO4, 100mg / L inositol, 0.3-0.5mg / L 6-BA, 0.1-0.2mg / L NAA, 30g / L sucrose, 8g / L agar powder, pH 5.8-6.2; The raw material components of the subculture culture medium in step (5) are: 1800 mg / L KNO3, 360 mg / L NH4NO3, 270 mg / L KH2PO4, 200 mg / L CaCl2, 178 mg / LCa(NO3)2, 360 mg / L MgSO4, 10 mg / L H3BO3, 25 mg / LMnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83 mg / L KI, 1000 mg / L PVP, 2.4 mg / L D-calcium pantothenate, 0.24 mg / L biotin, 100 mg / L L-cysteine, 2 mg / L glycine, 5 mg / L nicotinic acid, 2 mg / L thiamine, 0.5 mg / L pyridoxine, 37.3 mg / L Na2-EDTA, 27.8mg / L FeSO4, 100mg / L inositol, 0.3-0.5mg / L 6-BA, 0.2-0.4mg / L NAA, 30g / L sucrose, 8g / L agar powder, pH 5.8-6.2; The rooting medium raw material components of step (6) are: 900mg / L KNO3, 180mg / L NH4NO3, 135mg / LKH2PO4, 100mg / L CaCl2, 89mg / LCa(NO3)2, 180mg / L MgSO4, 10mg / L H3BO3, 25 mg / LMnSO4, 10 mg / L ZnSO4, 0.25 mg / L Na2MoO4, 0.025 mg / L CuSO4, 0.025 mg / L CoCl2, 0.83mg / L KI, 1000mg / L PVP, 2.4mg / L D-calcium pantothenate, 0.24mg / L biotin, 100mg / L L-cysteine, 2mg / L glycine, 5mg / L nicotinic acid, 2mg / L thiamine, 0.5mg / L pyridoxine, 37.3mg / L Na2-EDTA, 27.8mg / L FeSO4, 100mg / L inositol, 0.4-0.6mg / L IBA, 0.3-0.5mg / L ABT1, 30g / L sucrose, 8g / L agar powder, pH 5.8-6.2; The raw material components of the foliar nutrient solution described in step (7) are as follows: 1L of foliar nutrient solution contains 180mgKNO3, 36mgNH4NO3, 27mgKH2PO4, 20mgCaCl2, 36mgMgSO4, 1mgH3BO3, 2.5mgMnSO4, 1mgZnSO4, 0.025 mgNa2MoO4, 0.0025mgCuSO4, 0.0025mgCoCl2, and 0.083mgKI.

2. A method for raising seedlings of Eucalyptus eryngii according to claim 1, characterized in that: The sowing matrix described in step (1) is a mixture of loess soil and clean river sand in a weight ratio of 1:1, disinfected with a potassium permanganate solution with a volume concentration of 0.5%, and then covered with a film and sealed for one day. After the film is removed and aired, it is soaked with water and set aside.

3. A method for raising seedlings of Eucalyptus eryngii according to claim 1, characterized in that: Step (2) After the seedlings are transplanted, they are sprayed with thiophanate-methyl or chlorothalonil according to the conventional method every 7 days to prevent diseases. When the yellow-leaved eucalyptus seedlings grow new roots, they are sprayed with a compound fertilizer aqueous solution with a volume concentration of 0.2-0.5% every 10 days.

4. A method for raising seedlings of Eucalyptus eryngii according to claim 1, characterized in that: The transplanting medium described in step (2) and step (7) is a mixture of coconut husk, carbonized rice husk, peat soil and loess soil in a weight ratio of 6:2:1:1, and then adding 4kg / m3 of calcium magnesium phosphate fertilizer. 3 , covered with a film and piled up in the sun for a month to allow it to fully ferment.

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