Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the seedling hardening method of the tissue culture seedlings of the hybrid orchid, which can improve the seedling hardening survival rate of the tissue culture seedlings of the hybrid orchid and improve the potted plant quality.
To achieve the above object of the invention. The technical scheme of the invention is as follows:
a seedling hardening method for tissue culture seedlings of hybrid orchid comprises the procedures of tissue culture bottle seedling selection, small seedling treatment, small seedling cultivation, medium and large seedling transplantation and post-transplantation management, the tissue culture seedlings of hybrid orchid which grow robustly and have rooted roots are selected, cleaned, disinfected, the roots of the tissue culture seedlings are soaked in nutrient solution for 20-24 hours, the tissue culture seedlings are transplanted into a seedling culture container filled with a seedling culture medium to be cultivated for 4-6 months to obtain medium and large seedlings, the medium and large seedlings are transplanted into the culture medium on a cultivation field to be hardened, and the hybrid orchid is managed after watering, fertilizing and transplanting to obtain hybrid orchid seedlings; the main operation steps are as follows:
(1) selecting tissue culture bottle seedlings: selecting strong and rooted hybrid orchid tissue culture seedlings, placing the hybrid orchid tissue culture seedlings in a greenhouse, unscrewing a bottle cap to enable the tissue culture seedlings to adapt to the environment, taking out seedlings in the bottle after 5 days, cleaning a root culture medium, soaking roots in a disinfection solution for 10min, and airing until the roots turn white;
(2) and (3) treating the plantlets: wrapping the roots of the seedlings obtained in the step (1) with wet sphagna, and soaking the wrapped roots in a nutrient solution for 20-24h, wherein the soaking depth is 1/3 of root systems;
(3) and (3) seedling cultivation: transplanting the plantlets obtained in the step (2) into a seedling raising container with a seedling raising substrate, wherein the height of the seedling raising container is 8.5cm, the upper caliber of the seedling raising container is 4.8cm, the lower caliber of the seedling raising container is 2.0cm, and 50 holes are reserved in a greenhouse with the temperature of 20-25 ℃, the relative air humidity of 80-90% and the illumination intensity of 12000-15000 lx; 1-1.5g of solid slow release fertilizer with the effective components of N, P and K being 14, 13 and 13 is sprayed on each plant for 1 time every 3 months; watering is combined, simultaneously liquid water and fertilizer are applied every 10 days, and the cultivation is carried out for 4 to 6 months, so as to obtain seedlings with the medium and large diameters of 20 to 30 cm;
(4) transplanting medium and large seedlings: transplanting the medium and large seedlings obtained in the step (3) into a culture medium on a culture field for hardening seedlings, wherein the illumination intensity temperature in the culture field is 20-25 ℃, the air relative humidity is 75%, and the illumination intensity is 12000-;
(5) managing after transplanting: 1-1.5g of solid slow release fertilizer with the effective components of N, P and K being 14, 13 and 13 is sprayed on each plant for 1 time every 3 months; and (4) watering in combination, and spraying liquid water fertilizer once every 15 days until hybrid orchid seedlings are obtained.
Preferably, the disinfecting solution in step (1) is tea bran leach liquor with volume concentration of 0.03-0.05%.
Preferably, the nutrient solution in the step (2) consists of the following components in concentration: IAA 100-300 mg/L, ABT1100-150 mg/L paclobutrazol 10-15 mg/L, VB125-10 mg/L, biotin 50-100 mg/L, NH4NO3 400 mg/L 、KH2PO4400 mg/L、K2SO4200 mg/L、MgSO4•7H2O 400 mg/L、Ca(NO3)2100 mg/L、H3BO350 mg/L, EDTA-FeNa 10-15 mg/L, and adding citric acid solution and sodium hydroxide solution to adjust the pH value of the nutrient solution to 5.5-6.5.
Preferably, the seedling substrate in the step (3) is a mixture of cedar chips, spicy chips and eucalyptus chips according to a volume ratio of 1:1: 1; the seedling culture substrate is fermented by urea for more than half a year before use, and the culture substrate is disinfected by 2000 times of thiophanate methyl before cultivation.
Preferably, the culture medium in the step (4) is cedar wood chips; before use, the culture medium is fermented by urea for more than half a year, and before seedling, the seedling culture medium is disinfected by 2000 times of methyl mop body fluid.
Preferably, the preparation method of the liquid water fertilizer in the steps (3) and (5) comprises the following steps: 40-45 parts of urea, (NH) are weighed according to the weight4)2HPO4 30-35 parts of K2SO425-30 parts of the raw materials are sequentially added into 10000 parts of water and fully dissolved to obtain a mixed solution I; weighing MgSO450 portions of Ca (NO)3)250 portions of FeSO430 parts of MnSO430 portions of H3BO3 20 portions of ZnSO410 parts of CuSO410 parts of Na2MoO4Sequentially adding 10 parts of the mixture into 10000 parts of water, and fully dissolving to obtain a mixed solution II; uniformly mixing the mixed solution I and the mixed solution II according to the volume ratio of 500:1 to obtain a mixed solution III; and (3) adjusting the pH value of the mixed solution III to 5.8 by using 1 mol/L sodium hydroxide and hydrochloric acid solution to obtain the liquid water fertilizer.
Preferably, the seedling raising container in the step (3) is a seedling raising plug tray with the height of 8.5cm, the upper caliber of 4.8cm, the lower caliber of 2.0cm and 50 holes.
Compared with the prior art, the invention has the following advantages:
1. according to the growth characteristics of the hybrid orchid, different matrixes are adopted for carrying out seedling hardening in stages, so that the parameters of the orchid seedling such as newly added leaf number, new root increment, average root length increment, plant height increment and the like are high, the transplanting survival rate of the orchid hybrid tissue culture seedling is effectively improved, the pot culture quality is improved, and the economic benefit is better.
2. In the seedling cultivation stage, the cedar chips, the spicy chips and the eucalyptus chips are used as seedling cultivation substrates and contain various nutrient substances and trace elements, so that sufficient nutrients are provided for the orchid, the growth of the orchid is promoted, and the growth activity of the tissue culture seedling of the orchid is improved; in the transplanting stage of the medium and large seedlings, the cedar chips are used as the culture medium, the nutrition is rich, the growth requirement of orchid can be met, the contained aromatic substances can effectively prevent the breeding of bacteria, and the survival rate of the transplanted tissue culture seedlings is improved; the matrix adopted by the invention is derived from waste materials of wood processing, the waste of resources is avoided, the matrix has good water and air permeability, moisture and fertilizer are preserved, the transplanting survival rate reaches more than 95%, the growth speed is high, and the seedlings can grow in about one year.
3. The invention firstly immerses the root system of the seedlings in the tissue culture bottle into the nutrient solution before transplanting, directly absorbs the nutrients in the nutrient solution, improves the growth activity of the seedlings, effectively enhances the growth vigor of the seedlings after the tissue culture seedlings of the hybrid orchid are transplanted, and lays a foundation for ensuring the transplanting survival rate.
4. The liquid water fertilizer adopted after transplanting contains major elements and trace elements closely related to the growth of the orchid, the orchid seedlings can be quickly absorbed and utilized to promote the growth and become thick and strong after the liquid water fertilizer is applied, the uniformity of the orchid seedlings is high, and the quality is excellent.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example 1:
selecting strong and rooted hybrid orchid tissue culture seedlings of the cymbidium hybridum and the cymbidium hybridum, placing the hybrid orchid tissue culture seedlings in a greenhouse, unscrewing a bottle cap to enable the tissue culture seedlings to adapt to the environment, taking out seedlings in the bottle after 5 days, cleaning a root culture medium, soaking roots in a tea bran leaching liquor with the volume concentration of 0.03-0.05% prepared by a conventional method for 10min, and airing until the roots become white. The tea bran leaching liquor is obtained by soaking tea bran and warm water at 40 ℃ for 5 hours according to the weight ratio of 1: 5.
Cutting the plantlets with a moist mossThe roots are wrapped and then soaked in nutrient solution for 24 hours, and the soaking depth is 1/3 of the roots; the nutrient solution consists of the following components in concentration: IAA 100 mg/L, ABT1150 mg/L paclobutrazol 10 mg/L, VB1210 mg/L of biotin 50 mg/L, NH4NO3 400 mg/L 、KH2PO4400 mg/L、K2SO4200 mg/L、MgSO4•7H2O 400 mg/L、Ca(NO3)2100 mg/L、H3BO350 mg/L, EDTA-FeNa 10 mg/L, and adding citric acid solution and sodium hydroxide solution to adjust the pH value of the nutrient solution to 5.5.
Transplanting the soaked seedlings into seedling culture hole trays which are filled with seedling culture substrates and have the specification of height of 8.5cm, upper caliber of 4.8cm, lower caliber of 2.0cm and 50 holes, wherein the seedling culture substrates are mixture of cedar chips, peppery chips and eucalyptus chips according to the volume ratio of 1:1: 1; the seedling culture substrate is fermented by urea for more than half a year before use, and the culture substrate is disinfected by 2000 times of thiophanate methyl before cultivation. Placing the well-installed seedling culture hole tray in a greenhouse with the temperature of 20-25 ℃, the relative air humidity of 80-90% and the illumination intensity of 12000-; spraying a solid slow release fertilizer with the effective components of N, P and K being 14, 13 for 1 time every 3 months according to 1-1.5g of each plant after survival; watering is combined, simultaneously liquid water and fertilizer are applied every 15 days, and cultivation is carried out for 4-6 months, and medium and large seedlings with the length of 20-30cm are obtained.
Transplanting the obtained medium and large seedlings into a culture medium on a culture field for hardening seedlings, wherein the culture medium is cedar wood chips; before use, the culture medium is fermented by urea for more than half a year, and before seedling, the seedling culture medium is disinfected by 2000 times of methyl mop body fluid. The light intensity temperature in the cultivation field is 20-25 ℃, the air relative humidity is 75%, and the light intensity is 12000-. Spraying a solid slow release fertilizer with the effective components of N, P and K being 14, 13 and 13 for 1 time every 3 months according to 2-2.5g of each plant; and (4) watering in combination, and spraying liquid water fertilizer once every 15 days until hybrid orchid seedlings are obtained.
The preparation method of the liquid water fertilizer comprises the following steps: weighing 40 parts of urea and (NH) by weight4)2HPO4 35 parts of, K2SO425 parts of the raw materials are sequentially added into 10000 parts of water and fully dissolved to obtain a mixed solution I; weighing MgSO450 portions of Ca (NO)3)250 portions of FeSO430 parts of MnSO430 portions of H3BO3 20 portions of ZnSO410 parts of CuSO410 parts of Na2MoO4Sequentially adding 10 parts of the mixture into 10000 parts of water, and fully dissolving to obtain a mixed solution II; uniformly mixing the mixed solution I and the mixed solution II according to the volume ratio of 500:1 to obtain a mixed solution III; and (3) regulating the pH value of the mixed solution III to 5.8 by using 1 mol/L hydrochloric acid to obtain the liquid water fertilizer.
After 45 days of transplanting, the survival rate of the hybrid orchid of the Tonghai Jian multiplied by the cymbidium hybridum is 96.7 percent, the average number of newly added leaves is 2.5, the average increment of new roots is 2.4, the average increment of root length is 3.1cm, and the average increment of plant height is 2.7 cm.
Example 2:
selecting a strong and rooted cymbidium hybridum tissue culture seedling of cymbidium and small snow element, placing the tissue culture seedling in a greenhouse, unscrewing a bottle cap to enable the tissue culture seedling to adapt to the environment, taking out the small seedling in the bottle after 5 days, cleaning a root culture medium, soaking the root in a tea bran leaching liquor with the volume concentration of 0.04% prepared by a conventional method for 10min, and airing until the root is whitened. The tea bran leaching liquor is obtained by soaking tea bran and warm water at 40 ℃ for 5 hours according to the weight ratio of 1:5
Wrapping the roots of the seedlings with the wet sphagna, and soaking the seedlings in the nutrient solution for 22h, wherein the soaking depth is 1/3 of the roots; the nutrient solution consists of the following components in concentration: IAA 200 mg/L, ABT1130 mg/L paclobutrazol 12 mg/L, VB128 mg/L, biotin 80 mg/L, NH4NO3 400 mg/L 、KH2PO4400 mg/L、K2SO4200 mg/L、MgSO4•7H2O 400 mg/L、Ca(NO3)2100 mg/L、H3BO350 mg/L, EDTA-FeNa 12 mg/L, and adding citric acid solution and sodium hydroxide solution to adjust the pH value of the nutrient solution to 6.0.
Transplanting the soaked seedlings into seedling culture hole trays which are filled with seedling culture substrates and have the specification of height of 8.5cm, upper caliber of 4.8cm, lower caliber of 2.0cm and 50 holes, wherein the seedling culture substrates are mixture of cedar chips, peppery chips and eucalyptus chips according to the volume ratio of 1:1: 1; the seedling culture substrate is fermented by urea for more than half a year before use, and the culture substrate is disinfected by 2000 times of thiophanate methyl before cultivation. Placing the well-installed seedling culture hole tray in a greenhouse with the temperature of 20-25 ℃, the relative air humidity of 80-90% and the illumination intensity of 12000-; spraying a solid slow release fertilizer with the effective components of N, P and K being 14, 13 for 1 time every 3 months according to 1-1.5g of each plant after survival; watering is combined, simultaneously liquid water and fertilizer are applied every 15 days, and cultivation is carried out for 4-6 months, and medium and large seedlings with the length of 20-30cm are obtained.
Transplanting the obtained medium and large seedlings into a culture medium on a culture field for hardening seedlings, wherein the culture medium is cedar wood chips; before use, the culture medium is fermented by urea for more than half a year, and before seedling, the seedling culture medium is disinfected by 2000 times of methyl mop body fluid. The light intensity temperature in the cultivation field is 20-25 ℃, the air relative humidity is 75%, and the light intensity is 12000-. Spraying a solid slow release fertilizer with the effective components of N, P and K being 14, 13 and 13 for 1 time every 3 months according to 2-2.5g of each plant; and (4) watering in combination, and spraying liquid water fertilizer once every 15 days until hybrid orchid seedlings are obtained.
The preparation method of the liquid water fertilizer comprises the following steps: 42 portions of urea and (NH) are weighed according to the weight4)2HPO4 33 parts of, K2SO428 parts of the raw materials are sequentially added into 10000 parts of water and fully dissolved to obtain a mixed solution I; weighing MgSO450 portions of Ca (NO)3)250 portions of FeSO430 parts of MnSO430 portions of H3BO3 20 portions of ZnSO410 parts of CuSO410 parts of Na2MoO4Sequentially adding 10 parts of the mixture into 10000 parts of water, and fully dissolving to obtain a mixed solution II; uniformly mixing the mixed solution I and the mixed solution II according to the volume ratio of 500:1 to obtain a mixed solution III; and (3) regulating the pH value of the mixed solution III to 5.8 by using 1 mol/L hydrochloric acid to obtain the liquid water fertilizer.
After 45 days of transplanting, the survival rate of the hybrid cymbidium hybridum of cymbidium hybridum and small snowflake is 97.1 percent, the average number of newly added leaves is 2.6, the average increment of new roots is 2.3, the average increment of root length is 3.0cm, and the average increment of plant height is 2.9 cm.
Example 3:
selecting strong rooted hybrid orchid tissue culture seedlings of Wu-character emerald green and cymbidium hybridum, placing the hybrid orchid tissue culture seedlings in a greenhouse, unscrewing a bottle cap to enable the tissue culture seedlings to adapt to the environment, taking out seedlings in the bottle after 5 days, cleaning a root culture medium, soaking the roots in a tea bran leaching liquor with the volume concentration of 0.05% prepared by a conventional method for 10min, and airing until the roots become white. The tea bran leaching liquor is obtained by soaking tea bran and warm water at 40 ℃ for 5 hours according to the weight ratio of 1:5
Wrapping the roots of the seedlings with the wet sphagna, and soaking the seedlings in the nutrient solution for 20 hours, wherein the soaking depth is 1/3 of the roots; the nutrient solution consists of the following components in concentration: IAA 300 mg/L, ABT1100 mg/L paclobutrazol 15 mg/L, VB125 mg/L, biotin 100 mg/L, NH4NO3 400 mg/L 、KH2PO4400 mg/L、K2SO4200 mg/L、MgSO4•7H2O 400 mg/L、Ca(NO3)2100 mg/L、H3BO350 mg/L, EDTA-FeNa 15 mg/L, and adding citric acid solution and sodium hydroxide solution to adjust the pH value of the nutrient solution to 6.5.
Transplanting the soaked seedlings into seedling culture hole trays which are filled with seedling culture substrates and have the specification of height of 8.5cm, upper caliber of 4.8cm, lower caliber of 2.0cm and 50 holes, wherein the seedling culture substrates are mixture of cedar chips, peppery chips and eucalyptus chips according to the volume ratio of 1:1: 1; the seedling culture substrate is fermented by urea for more than half a year before use, and the culture substrate is disinfected by 2000 times of thiophanate methyl before cultivation. Placing the well-installed seedling culture hole tray in a greenhouse with the temperature of 20-25 ℃, the relative air humidity of 80-90% and the illumination intensity of 12000-; spraying a solid slow release fertilizer with the effective components of N, P and K being 14, 13 for 1 time every 3 months according to 1-1.5g of each plant after survival; watering is combined, simultaneously liquid water and fertilizer are applied every 15 days, and cultivation is carried out for 4-6 months, and medium and large seedlings with the length of 20-30cm are obtained.
Transplanting the obtained medium and large seedlings into a culture medium on a culture field for hardening seedlings, wherein the culture medium is cedar wood chips; before use, the culture medium is fermented by urea for more than half a year, and before seedling, the seedling culture medium is disinfected by 2000 times of methyl mop body fluid. The light intensity temperature in the cultivation field is 20-25 ℃, the air relative humidity is 75%, and the light intensity is 12000-. Spraying a solid slow release fertilizer with the effective components of N, P and K being 14, 13 and 13 for 1 time every 3 months according to 2-2.5g of each plant; and (4) watering in combination, and spraying liquid water fertilizer once every 15 days until hybrid orchid seedlings are obtained.
The preparation method of the liquid water fertilizer comprises the following steps: weighing 45 parts of urea and (NH) by weight4)2HPO4 30 parts of, K2SO430 parts of the raw materials are sequentially added into 10000 parts of water and fully dissolved to obtain a mixed solution I; weighing MgSO450 portions of Ca (NO)3)250 portions of FeSO430 parts of MnSO430 portions of H3BO3 20 portions of ZnSO410 parts of CuSO410 parts of Na2MoO4Sequentially adding 10 parts of the mixture into 10000 parts of water, and fully dissolving to obtain a mixed solution II; uniformly mixing the mixed solution I and the mixed solution II according to the volume ratio of 500:1 to obtain a mixed solution III; and (3) regulating the pH value of the mixed solution III to 5.8 by using 1 mol/L hydrochloric acid to obtain the liquid water fertilizer.
After 45 days of transplantation, the survival rate of hybrid orchid of Wu-character cui-X cymbidium hybridum is 98.0 percent, the average number of newly added leaves is 2.5, the average new root increment is 2.5, the average root length increment is 3.3cm, and the average plant height increment is 2.6 cm.