A kind of method for rapid propagation and seedling growth of flue-cured tobacco
A technology for rapid seedling breeding and tissue culture, which is applied in the field of tobacco production to achieve the effects of prospering the rural economy, increasing the yield of tobacco leaves, and reducing the incidence of insect pests
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Embodiment 1
[0030] Embodiment 1 Tobacco Seedling Acquisition
[0031] Wrap the seeds of tobacco variety K326 (provided by Guangdong Tobacco Qingyuan City Co., Ltd.) with gauze, rinse them in running water for 10 to 15 minutes, and then soak the seeds in 75% ethanol solution for 30 seconds on an ultra-clean workbench. Rinse 3 times; then disinfect in 0.1% HgCl solution for 5-7 minutes; rinse 5 times with sterile water; sow in 1 / 2MS medium. Germination culture was carried out under (26±1)°C and 16h / d light conditions. Then subculture, in the subculture medium: MS+0.5mg / L naphthaleneacetic acid (NAA)+1.0mg / L6-benzylaminopurine (6-BA)+7.5~8.0g / L kara powder+0.05g / Differentiate buds on L-inositol + 30g / L sucrose; then root and strengthen seedling medium B: 1 / 2MS + 7.5-8.0g / L kara powder + 0.2mg / LNAA + 0.05g / L inositol + 30g / L of sucrose+0.5g / L activated carbon (AC) to take root and grow strong seedlings into required aseptic seedlings.
Embodiment 2
[0032] Induction of embodiment 2 callus
[0033] Take aseptic seedlings with a leaf age of about 3, cut the leaves into 0.5cm*0.5cm size, sow them in the 16 kinds of media in the following table 1, and connect 8 bottles for three replicates for each treatment, and inoculate 5 pieces in each bottle , cultured at (26±1)°C, dark conditions until callus appeared, and then moved to (26±1)°C, 16h / d light conditions for cultivation. Observe and record every five days, and study the callus morphology, growth speed, and induction rate. The specific results are shown in Table 1.
[0034] Table 1 Effects of different hormone concentration ratios on callus growth and induction rate
[0035]
[0036] As can be seen from the data in Table 1, under the culture conditions of the present invention, the medium selection formula for inducing callus production is MS+Kara powder 7.5~8.0g / L+inositol 0.05g / L+sucrose 30g / L+0.2 mg / L naphthaleneacetic acid (NAA) + 2.0mg / L kinetin (KT), the callus ...
Embodiment 3
[0037] The differentiation subculture of embodiment 3 buds
[0038] During the bud differentiation, the 4 formulations with the best effect in the callus induction were taken, and then the shoots were subcultured. Each treatment received 8 bottles for three replicates, and each bottle was inoculated with 4 evenly cut shoots.
[0039] The bud differentiation subculture formula adopts several formulas that perform better in callus induction. The results of different concentrations of hormone ratios on bud differentiation are as follows:
[0040] From Table 2 and figure 1 It can be seen that among the various formulas tested, the value-added rate of A3 buds has reached 100%, the pollution rate is low, and the effect of differentiation and subculture is good. The buds of A3 are in the best shape, and the buds are relatively clustered. Long roots, long seedlings.
[0041] Table 2 Effects of different hormone concentration ratios on bud differentiation value-added rate and bud poi...
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