CN-121986709-A - Cultivation method of root-free and leaf-free ginger tissue culture seedlings

CN121986709ACN 121986709 ACN121986709 ACN 121986709ACN-121986709-A

Abstract

The invention provides a cultivation method of a root-free and leaf-free ginger tissue culture seedling, which relates to the field of seed seedling cultivation and comprises the steps of material preparation, mixed matrix preparation and tissue culture seedling field planting, wherein the tissue culture seedling field planting comprises the steps of enabling a root-free and leaf-free tissue culture seedling to be vertically mixed into an implantation hole of the mixed matrix, ensuring that a root neck part is completely embedded into the matrix hole, stem sections are vertical, lightly compacting the matrix around a seedling plant, enabling the seedling plant to be fixed and free of lodging, and finally carrying out four-stage cultivation according to time sequence to obtain a transplanted tissue culture seedling. The method cultivates ginger plant groups with consistent tillering and balanced structure through transplanting tissue culture seedlings without roots and leaves and reconstructing root systems with new roots, thereby improving the adaptability and survival rate after transplanting.

Inventors

JIANG LIHUI
PENG KE
Ran Xingrong
YU LI
CHEN LAN
LI HONGLEI
SONG CHAO
XIA MAOQIN

Assignees

重庆文理学院

Dates

Publication Date: 20260508
Application Date: 20260403

Claims (10)

1. A cultivation method of a root-free and leaf-free ginger tissue culture seedling is characterized by comprising the following steps: step S1, preparing materials, namely cutting off all callus of original root hairs and basal parts of ginger detoxification tissue culture seedlings, only reserving root neck parts connecting overground parts and root systems, simultaneously cutting off overground parts and overground long stems and all leaves, only reserving healthy stem segments of 2-3 cm, and enabling incisions to be flat and free from tearing; s2, preparing a mixed matrix in advance, and filling the mixed matrix into a container after nondestructive sterilization to make the surface of the mixed matrix flat; S3, field planting of tissue culture seedlings, namely selecting a seedling raising tray with a water absorption hole at the bottom and a matched tray as a field planting container, uniformly filling mixed matrixes into each hole position of the seedling raising tray, slightly scraping and slightly compacting the mixed matrixes to the position 1cm above the hole position, then vertically punching holes in the center of each hole position matrix, enabling the hole depth to be 0.5-1 cm, enabling the hole diameter to be matched with a stem section of the tissue culture seedlings, then vertically planting the root-free and leaf-free tissue culture seedlings obtained in the step S1 into the holes, ensuring that the neck parts of roots are completely embedded into the holes of the matrixes, enabling the stem sections to be vertical, slightly compacting the matrixes around the seedling plants, enabling the seedling plants to be fixed and free from lodging, and finally sequentially carrying out four-stage cultivation according to time to obtain the transplanted tissue culture seedlings.
2. The cultivation method of the ginger tissue culture seedling without roots and leaves of claim 1, which is characterized in that in the step S1, the ginger detoxification tissue culture seedling is selected to grow for 30-45 days, the height of the seedling is 5-8 cm, 3-5 tissue culture seedlings with fully-unfolded leaves, strong stems, strong growth, no vitrification and obvious plant diseases and insect pests are grown, and the ginger detoxification tissue culture seedling is taken out of a culture bottle and rinsed for 3-4 times by sterile clean water.
3. The method for cultivating a root-free and leaf-free ginger tissue culture seedling according to claim 1 or 2, wherein in the step S2, the mixed matrix comprises hydrophobically modified expanded perlite microspheres, amino modified silica aerogel particles, a bacteria-carrying humic acid grafted vermiculite composite carrier and enzymatic peat fibers, and the mass percentages of the components are 29.5-30.5%, 8.5-9.5%, 37.5-38.5% and 22.5-23.5%, respectively.
4. A cultivation method of a rootless and foliar-less ginger tissue culture seedling according to claim 2 or 3 is characterized in that the preparation method of the hydrophobically modified expanded perlite microsphere specifically comprises the steps of washing 30-50 meshes of perlite, drying, immersing in KH 570-absolute ethanol solution with the concentration of 1.8-2.2%, wherein the mass volume ratio of the perlite to the solution is 0.9-1.1 g:4.5-5.5 mL, oscillating for 3.8-4.2 h at the speed of 120-150 rpm at 38-42 ℃, filtering, washing, and drying for 2-3 h at the temperature of 102-108 ℃.
5. A cultivation method of a rootless and foliar-free ginger tissue culture seedling is characterized in that the preparation method of the amination modified silica aerogel particles comprises the steps of vacuum drying 100-200 meshes of silica aerogel, immersing the silica aerogel in KH 550-absolute ethanol solution with the concentration of 2.8-3.2%, wherein the mass volume ratio of the silica aerogel to the solution is 0.9-1.1 g:3.6-4.4 mL, oscillating for 5.8-6.2 h at a rotating speed of 100-130 rpm under the condition of avoiding light at 48-52 ℃, filtering and cleaning, and vacuum drying for 4-6 h at 58-62 ℃ and minus 0.08-0.09 MPa.
6. The cultivation method of the rooting-free and foliar-free ginger tissue culture seedling is characterized by comprising the steps of adding 200-mesh vermiculite into hydrochloric acid solution with the concentration of 1mol/L, stirring the vermiculite and the hydrochloric acid solution for 3.8-4.2 h at the temperature of 78-82 ℃ at the rotating speed of 140-160 rpm, filtering and washing to be neutral, and drying at the temperature of 102-108 ℃ for 2-3 h; mixing the activated vermiculite and fulvic acid according to a mass ratio of 4.75-5.25:1.9-2.1, adding a mixed cross-linking agent of EDC and NHS, wherein the molar ratio of EDS to NHS is 1:1, the mixed cross-linking agent accounts for 0.48-0.52% of the total mass of the vermiculite and fulvic acid, stirring for 5.8-6.2 h at a constant temperature water bath of 48-52 ℃ at a rotating speed of 180-220 rpm, filtering and cleaning, vacuum drying for 3-5 h at 58-62 ℃ and-0.08 MPa for 3-5 h at-0.09 MPa to obtain a carrier, finally, placing the sterilized carrier into a sterile vacuum adsorption bottle, adding a candida langbei bacterial suspension with a bacterial liquid concentration of 1X 10 9 CFU/mL, keeping the solid-liquid ratio of the carrier to the bacterial suspension at 0.9-1.1 g:2.7-3.3 mL, slowly introducing the carrier into the carrier under a vacuum degree of 180-220 MPa, and performing air permeation under a pressure difference of the carrier to obtain the carrier, and performing air drying for 10-30 ℃ after the carrier is cooled.
7. A cultivation method of a rootless and foliar-free ginger tissue culture seedling is characterized in that peat is crushed, acetic acid solution with pH of 4.8 and sodium acetate solution are added into buffer solution mixed to form the peat tissue culture seedling, the mass volume ratio of peat to the buffer solution is 0.9-1.1 g:9-11 mL, the peat suspension is uniformly stirred at a rotating speed of 180-220 rpm to form peat suspension, cellulase is added into the peat suspension, the cellulase is in an amount of 1.8-2.2% of peat mass, the cellulase is stirred at a rotating speed of 140-160 rpm for 3.8-4.2 h at 48-52 ℃, then the peat suspension after enzymolysis is placed into constant-temperature waste water bath with the pH of 98-102 ℃, the temperature is kept for 8-10 min, the peat fiber is repeatedly washed by deionized water until the pH of filtrate is stabilized at 5.5-6.0, finally, a fiber filter cake is collected, air-dried, classified, and the classified fiber is dried to constant weight at 58-62 ℃ after classification.
8. The method for cultivating a tissue culture seedling of ginger without roots and leaves according to claim 3, wherein in the step S3, four stages comprise a root primordium differentiation cultivation stage, an adventitious root construction cultivation stage, a root system stress-resistant tillering cultivation stage and a strong seedling cultivation stage, wherein the root primordium differentiation cultivation stage is 1-7 days after field planting, the adventitious root construction cultivation stage is 8-20 days after field planting, the root system stress-resistant tillering cultivation stage is 21-35 days after field planting, and the strong seedling cultivation stage is 36-40 days after field planting.
9. The cultivation method of the rootless and foliar-free ginger tissue culture seedlings according to claim 8, wherein the rootlet differentiation cultivation stage is cultivated in an environment with the temperature of 24-26 ℃ and the air humidity of not less than 80% RH, the illumination intensity of 2400-2600 Lux and the illumination time of 12h/d, and the rootlet differentiation nutrient solution is adopted for irrigation, namely, the rootlet differentiation nutrient solution is adopted for primary irrigation after the field planting is finished, the substrate is adopted for one-time watering, and the excessive effusion in a tray is poured off; The root primordium differentiation nutrient solution comprises 1/10 MS culture medium, pH sensitive chitosan coating combined amino acid nitrogen, N- (1-naphthyl) o-carbamyl benzoic acid, 24-epibrassinolide, 1, 4-butanediamine, tea tree essential oil nanoemulsion, carbamide peroxide, gamma-polyglutamic acid and potassium fulvate, wherein the concentration of the pH sensitive chitosan coating combined amino acid nitrogen, N- (1-naphthyl) o-carbamyl benzoic acid, 24-epibrassinolide, 1, 4-butanediamine, tea tree essential oil nanoemulsion, carbamide peroxide, gamma-polyglutamic acid and potassium fulvate is respectively 0.78～0.82g/L、0.09～0.11μmol/L、0.18～0.22μmol/L、0.9～1.1μmol/L、0.09～0.11g/L、4.8～5.2mg/L、0.28～0.32g/L、0.18～0.22g/L.
10. The cultivation method of the rootless and foliar-free ginger tissue culture seedlings according to claim 8, wherein the adventitious root construction cultivation stage is cultivated in an environment with the temperature of 24-26 ℃, the air humidity of 75% -80% RH, the illumination intensity of 3400-3600 Lux and the illumination time of 14h/d, and the adventitious root construction nutrient solution is adopted for irrigation, namely, the first irrigation at 8 th day after field planting, the adventitious root construction nutrient solution is adopted for one-time matrix pouring, and the excessive accumulated liquid in a tray is poured out, and the subsequent supplementary irrigation is carried out for 1 time every 5 days, wherein the water holding capacity of each irrigation is 60% of the matrix; The adventitious root constructing nutrient solution comprises 1/10 MS culture medium, N6-isopentenyl adenosine, L-tryptophan, strigolactone analogues, sodium borate, zinc sulfate, salicin, humic acid chelated compound trace elements, gamma-polyglutamic acid, chitosan oligosaccharide and bacillus subtilis, wherein the concentration of the N6-isopentenyl adenosine, L-tryptophan, strigolactone analogues, sodium borate, zinc sulfate, salicin, humic acid chelated compound trace elements, gamma-polyglutamic acid and chitosan oligosaccharide is 0.58～0.62μmol/L、0.08～0.12μmol/L、0.28～0.32μmol/L、1.9～2.1μmol/L、0.08～0.12μmol/L、0.48～0.52μmol/L、0.95～1.05mL/L、0.28～0.32g/L、0.33～0.37g/L; and the concentration of the bacillus subtilis is 2X 10 8 CFU/mL respectively.

Description

Cultivation method of root-free and leaf-free ginger tissue culture seedlings Technical Field The invention relates to the technical field of seed seedling cultivation, in particular to a cultivation method of a root-free and leaf-free ginger tissue culture seedling. Background In recent years, the detoxification technology of ginger is mature and perfect, the tissue culture seedling of ginger is gradually applied to production, however, compared with the mode of propagation through traditional tubers, the tissue culture seedling of ginger still has the defect of more complex domestication and transplanting processes. The method is characterized by comprising the steps of 1, weak physiological basis and poor adaptability, wherein root systems of ginger tissue culture seedlings are tender, regeneration capability is weak, most of the ginger tissue culture seedlings are adventitious roots, functions of absorbing moisture and nutrients are imperfect, water loss and wilting are easy to occur after transplanting, meanwhile, the ginger tissue culture Miao Qikong is low in potassium content of guard cells, cannot be autonomously synthesized into enough nutrients, depends on external supply and grows slowly, and in addition, tissue culture Miao Chuqi is easy to be infected by bacteria and die and is extremely sensitive to environmental fluctuation (temperature, humidity and illumination). 2. The physiological state of the tissue culture seedlings induced in the bottle is different from that of field plants, the bottle is in a stable environment with constant temperature and humidity, sterility and weak light, and the tissue culture seedlings need to adapt to the fluctuation of field temperature and humidity, the competition of microorganisms and the change of illumination intensity after transplanting, so that the physiological regulation difficulty is high. 3. The method has the advantages of complex operation flow and high cost, and needs to be subjected to multiple links such as uncapping seedling hardening, culture medium cleaning, disinfection, transplanting, seedling recovery management and the like, the links are numerous, manual work is relied on, the domestication period is long, the cultivation period is long, and the artificial influence factors are high. 4. The survival rate and the growth performance are limited, the overall survival rate is generally 60% -80% and is far lower than that of the traditional separated seedlings, the survival seedlings are slow in seedling returning speed, weak in early growth vigor and delayed in growth period, the expansion and the final yield of ginger rootstock are affected, and the problems of character separation, quality reduction and the like are easy to occur after transplanting. In addition, the problem of contradiction between root development and yield of the plants propagated by the ginger tissue culture seedlings is that the root number of the ginger tissue culture seedlings can reach 2.21 times that of the plants propagated by the traditional ginger blocks, but the proportion of endogenous hormones (for example, the IAA/ZR ratio is abnormally increased) is unbalanced, and the normal expansion of underground ginger blocks is inhibited, so that the root system is particularly developed, but the ginger blocks are small and the yield is low. In addition, when the tissue culture seedlings are transplanted to the soil environment, severe environmental stress is caused, so that the seedling recovery period is long, the survival rate is unstable, and the production risk and the cost of the tissue culture seedlings are further improved. Disclosure of Invention Aiming at the problems existing in the prior art, the invention aims to provide a cultivation method of a root-free and leaf-free ginger tissue culture seedling, which is characterized in that a ginger plant group with consistent tillering and balanced structure is cultivated by transplanting the root-free and leaf-free tissue culture seedling and reconstructing a root system, so that the adaptability and the survival rate after transplanting are improved. The aim of the invention is achieved by the following technical scheme: A cultivation method of a root-free and leaf-free ginger tissue culture seedling comprises the following steps: step S1, preparing materials, namely cutting off all callus of original root hairs and basal parts of ginger detoxification tissue culture seedlings, only reserving root neck parts connecting overground parts and root systems, simultaneously cutting off overground parts and overground long stems and all leaves, only reserving healthy stem segments of 2-3 cm, and enabling incisions to be flat and free from tearing; s2, preparing a mixed matrix in advance, and filling the mixed matrix into a container after nondestructive sterilization to make the surface of the mixed matrix flat; S3, field planting of tissue culture seedlings, namely selecting a seedling raising tray with a water