Regeneration and propagation method for optimized fresh ginger virus-free seedlings and development of industrialized flow of optimized fresh ginger virus-free seedlings
A technology for detoxifying seedlings and ginger, applied in the field of plant tissue culture, can solve the problems of easy-to-proliferate bacteria, low reproduction efficiency, poor growth after multiple subcultures, etc., and achieves the effects of short cycle, high reproduction efficiency and good state.
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Embodiment 1
[0029] The selection of embodiment 1, ginger explant
[0030] Select ginger pieces that are fat, plump, bright in skin color, fresh in flesh, not dry or shrink, not rotten, not frozen, and hard in texture, and accelerate germination at 28°C and 45% humidity for about 30 days, until the ginger buds grow to 1 ~2cm shoots are cut off for later use.
[0031] The shape of ginger explants directly affects the germination rate and growth strength. The traditional method does not strictly select the explants, which will cause the germination rate and growth of offspring ginger seedlings to be uneven. In this study, the selection of ginger explants was explored. The specific method is as follows:
[0032] (1) Select the ginger buds cultivated above, process with 75% alcohol for 1min, 0.1% mercuric chloride for 10min, wash with sterile water for 3-5 times, peel off the leaf primordia, and obtain 20 ginger bud explants in different states (such as figure 1 shown);
[0033] (2) Place...
Embodiment 2
[0037] Embodiment 2, ginger explant disinfection condition optimization
[0038] Ginger explants will carry a variety of bacteria and fungi, and incomplete disinfection will reduce the overall germination rate of ginger explants and affect the growth of offspring. In the traditional method, the use of less disinfectant dose will cause high pollution rate, and high disinfectant dose will cause low germination rate. In this study, different disinfectant combinations were used, different disinfection conditions were optimized, and a disinfection method with low contamination rate and high germination rate was identified.
[0039] (1) Clean the ginger buds with tap water after peeling them off, then remove the outermost skin, and carry out disinfection treatment in the ultra-clean workbench;
[0040] (2) Different disinfection conditions are as follows:
[0041] Disinfection method 1: 75% alcohol treatment for 1 minute, 0.1% mercury chloride treatment for 10 minutes, and sterile...
Embodiment 3
[0049] Embodiment 3, the selection of induction medium hormone kind
[0050] The germination rate of ginger explants and the differentiation rate of offspring are affected by the types and concentrations of hormones. In this study, the types of cytokinins were screened.
[0051] (1) Cleaning with tap water after peeling off the ginger buds, then removing the outermost epidermis, carrying out disinfection treatment (disinfection method 2) in the ultra-clean workbench, and then peeling off the leaf primordia to obtain the C state explants (see figure 1 ) 60 in total, divided into 3 groups on average;
[0052] (2) Prepare the induction medium of the following different cytokinins and carry out the induction treatment on the explants;
[0053] Medium 1: MS medium + 2mg / LTDZ + 0.5mg / LNAA + 8g agar;
[0054] Medium 2: MS medium + 2mg / LKT + 0.5mg / LNAA + 8g agar;
[0055] Medium three: MS medium + 2mg / L6-BA + 0.5mg / LNAA + 8g agar;
[0056] (3) The culture condition was 26°C and li...
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