Tissue culture rapid propagation method for limacia sagittata

A technology of tissue culture rapid propagation and green ox gall, applied in horticultural methods, botany equipment and methods, horticulture, etc., can solve the problems of low seed reproduction rate, long germination cycle, low seed germination rate, etc., and achieve high industrial production The effect of promotion value, fast reproduction speed and high tissue culture efficiency

Active Publication Date: 2018-12-07
INST OF MEDICINAL PLANTS YUNNAN ACAD OF AGRI SCI +1
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AI-Extracted Technical Summary

Problems solved by technology

[0003] The medicinal herb Lavender, which uses Tinobilis as the plant-based source, is a cold-backed medicinal material. However, in recent years, with the research, development and application of uses, the demand for Lavender has increased by leaps and bounds, and the limited wild resources cannot meet the market demand. , its price has also risen rapidly thereupon, and various enterprises and institutions have carried out planting technology research on green bile gall, but due to the small amount of green cattle gallbladder seeds, low seed ger...
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Abstract

The invention discloses a tissue culture rapid propagation method for limacia sagittata. The method comprises (1) explant selection; (2) explant disinfection; (3) induction of cluster buds; (4) proliferation of the cluster buds; and (5) serial rooting steps of the cluster buds. The efficient and low-cost tissue culture rapid propagation method for limacia sagittata is established, so that the scaled planting of limacia sagittata becomes possible.

Application Domain

Horticulture methodsPlant tissue culture

Technology Topic

BudCell budding +1

Image

  • Tissue culture rapid propagation method for limacia sagittata

Examples

  • Experimental program(2)
  • Comparison scheme(2)

Example Embodiment

[0020] Example 1
[0021] (1) the robust plants are planted indoors, sprayed 3 times with 500 times of 75% carbendazim every 7 days, and the young stem segments of 20~30cm at the ends are selected as explants;
[0022] (2) Defoliate the selected explants, cut them into 2-3cm stem sections with buds, rinse with saturated soapy water solution, use a soft toothbrush to remove dirt from the epidermis and depressions, and rinse with tap water for 3-4 When there is no soap residue, add 2 to 3 drops of Tween-80 and an appropriate amount of water, shake for 15 minutes, and rinse with tap water for 60 minutes. Transfer to the ultra-clean workbench, disinfect with 75% alcohol for 40s, rinse with sterile water for 2 to 3 times, disinfect with saturated bleach solution for 40 minutes, rinse with sterile water for 2 to 3 times, 0.1% mercuric solution + 2 to 3 drops of vomiting Sterilize at -80 temperature for 15min, rinse with sterile water 6 times;
[0023] (3) Use sterile paper to absorb the surface moisture of the sterilized explants, cut the wounds at both ends to 1.0-1.5cm of the stem section, and insert them obliquely into WPM+IAA 0.1mg/L+KT 3.0mg/L+TDZ On the induction medium of 0.2mg/L + charcoal 0.5g/L, at a temperature of 26 °C, with a light intensity of 2000lx, a single day of light time of 12h, the light and dark were alternately cultured for 50 days; the pollution rate and the induction rate of cluster buds were recorded. .
[0024] (4) Cut the induced clump buds into single buds and insert them obliquely into the proliferation culture of MS+NAA 0.1mg/L+KT 3.0mg/L+TIBA2.0mg/L+silver nitrate 1.0μg/L+coconut juice 100ml/L In the base, the temperature was 26°C, the light intensity was 2000lx, and the light-dark time was 12h in a single day for 33 days; the proliferation of the clump buds was recorded.
[0025] (5) Cut the induced or proliferated cluster buds into stem segments with 2 to 3 buds, stand upright in the rooting medium of 1/2MS+IBA 2.0mg/L+coconut juice 100ml/L+activated carbon 0.5g/L, in The temperature was 26 °C, the light intensity was 4000 lx, and the light and dark were alternately cultivated for 12 h on a single day, and the cultivation was carried out for 35 days; the rooting situation was recorded.
[0026] (6) with 2 to 3 roots and a height of 5 to 10 cm in a tissue culture bottle of green ox gallbladder, wash the culture medium, and plant it in a 5:1 ratio of peat soil and perlite. Keep the air humidity at 80%, carry out hardening; record the situation of hardening.

Example Embodiment

[0027] Example 2
[0028] (1) the robust plant is planted in the room, sprayed 4 times with 500 times of 75% carbendazim every 7 days, and the young stem section of 20~30cm at the end is selected as the explant;
[0029] (2) Defoliate the selected explants, cut them into 2-3cm stem sections with buds, rinse with saturated soapy water solution, use a soft toothbrush to remove dirt from the epidermis and depressions, and rinse with tap water for 3-4 When there is no soap residue, add 2 to 3 drops of Tween-80 and an appropriate amount of water, shake for 10 minutes, and rinse with tap water for 50 minutes. Transfer to the ultra-clean workbench, disinfect with 75% alcohol for 30s, rinse with sterile water for 2 to 3 times, disinfect with saturated bleach solution for 30 minutes, rinse with sterile water for 2 to 3 times, 0.1% mercuric solution + 2 to 3 drops of spit Sterilize at temperature -80 for 10 minutes, rinse with sterile water for 7 times;
[0030] (3) Use sterile paper to absorb the surface moisture of the sterilized explant, cut the wounds at both ends to 1.0-1.5cm of the stem, and insert it obliquely into WPM+IAA 0.01mg/L+KT 2.0mg/L+TDZ On the induction medium of 0.05mg/L + charcoal 0.5g/L, at a temperature of 23°C, with a light intensity of 1000lx, a single day of light time of 12h, the light and dark were alternately cultured for 58 days; the pollution rate and the induction rate of cluster buds were recorded. .
[0031] (4) Cut the induced clump buds into single buds and insert them obliquely into the proliferation culture of MS+NAA 0.01mg/L+KT 2.0mg/L+TIBA1.0mg/L+silver nitrate 1.0μg/L+coconut juice 50ml/L In the base, the temperature was 23°C, the light intensity was 1000lx, and the light-dark time was 12h in a single day for 40 days; the proliferation of the clump buds was recorded.
[0032] (5) Cut the induced or proliferated cluster buds into stem segments with 2 to 3 buds, and stand upright in the rooting culture of 1/2MS+IBA 1.0mg/L+coconut juice 50ml/L+activated carbon 0.5g/L. The temperature was 23 °C, the light intensity was 3000 lx, and the light and dark were alternately cultivated for 12 h on a single day, and the cultivation was carried out for 42 days; the rooting situation was recorded.
[0033] (6) with 2 to 3 roots and a height of 5 to 10 cm in a bottle of green ox gallbladder tissue culture seedlings, wash the culture medium, and plant them in a 5:1 ratio of peat soil and perlite. Keep the air humidity at 80%, carry out hardening; record the situation of hardening.

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