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Stevia rebaudiana tissue culture method and culture medium thereof

A tissue culture, stevia technology, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of low survival rate, quality degradation, long time, etc. easy effect

Inactive Publication Date: 2014-05-07
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method of seed propagation has a low survival rate; and the cutting propagation leads to quality degradation, and these two propagation methods take a long time and require a large amount of materials such as stem tips, stem segments with axillary buds, and seeds.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Under aseptic conditions, take the green leaves of Stevia rebaudiana stem tissue culture regeneration aseptic seedlings, contact the culture medium with the complete blade front, and inoculate the thickening induction medium on the ultra-clean workbench; thickening induction culture The base composition is MS+benzyladenine 6-BA 0.4mg / L+naphthaleneacetic acid NAA 0.1mg / L+0.7% agar;

[0032] (2) After being cultivated in a sterile culture room for 9 days, the leaves appeared fleshy and thickened, and were transferred to the germination induction medium under sterile conditions; the composition of the germination induction medium was MS + naphthalene acetic acid NAA 0.5mg / L + benzyl adenine 6-BA 2.0mg / L+0.7% agar;

[0033] (3) Start light culture after 1 week of shading and dark culture in a sterile culture room. After 12 days, the leaves germinate and appear purple buds on both sides of the midrib. The number of buds per explant is 13±2. After the seedling height is ...

Embodiment 2

[0039] (1) Pick the leaves from outdoor cultivated plants, wash them with running water for 30 minutes, treat them with 75% alcohol for 6-8 seconds on the ultra-clean workbench, then soak them with 0.1% mercury liter for 8 minutes, pour off the mercury chloride solution, and use Rinse with sterile water 4 to 5 times, not less than half a minute each time, then blot the surface water with sterile paper and avoid overlapping each other, and inoculate it into the thickening induction medium as soon as possible. The composition of the thickening induction medium is MS+benzyl Base adenine 6-BA 0.4mg / L+ naphthalene acetic acid NAA 0.1mg / L+0.7% agar;

[0040] (2) After being cultivated in a sterile culture room for 9 days, the flesh thickened and transferred to the germination induction medium under sterile conditions. BA 2.0mg / L+0.7% agar;

[0041] (3) After 1 week of shading and dark culture in a sterile culture room, light culture was started. After 14 days, purple buds germinate...

Embodiment 3

[0048] (1) Pick the leaves from outdoor cultivated plants, wash them with running water for 30 minutes, treat them with 75% alcohol for 6-8 seconds on the ultra-clean workbench, then soak them with 0.1% mercury liter for 8 minutes, pour off the mercury chloride solution, and use Rinse with sterile water 4 to 5 times, not less than half a minute each time, then blot the surface water with sterile paper and avoid overlapping each other, and inoculate it into the thickening induction medium as soon as possible. The composition of the thickening induction medium is MS+benzyl Base adenine 6-BA 0.5mg / L+ naphthalene acetic acid NAA 0.1mg / L+0.7% agar;

[0049] (2) After being cultivated in a sterile culture room for 9 days, the flesh thickened, and transferred to the germination induction medium under sterile conditions. BA 2.5mg / L+0.7% agar;

[0050] (3) After 1 week of shading and dark culture in a sterile culture room, light culture was started. After 14 days, purple buds germinat...

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Abstract

The invention relates to a stevia rebaudiana tissue culture propagation method and a culture medium of the method, and particularly relates to a tissue culture rapid propagation method for directly forming a bud by using a stevia rebaudiana leaf as an explant. The stevia rebaudiana tissue culture method comprises the following steps: (1), collecting the explant; (2), disinfecting and inoculating; (3), performing thickening induction-culturing; (4), budding and seedling; (5), transferring and grafting, and rooting; and (6), hardening and transplanting. According to the stevia rebaudiana tissue culture method provided by the invention, materials are easily available, a plurality of seed sources can be rapidly propagated, the explant tissue culture has no culture phase of callus cell proliferation and differentiation, thus the culture time is shortened, extremely high reproduction rate is achieved; the stevia rebaudiana tissue culture propagation method is easy to operate and is capable of realizing large-scale production; and the operation of researches such as genetic transformation is facilitated in a culture process. The culture medium provided by the invention ensures that the leaf is capable of reaching the expected culture target in each phase.

Description

technical field [0001] The invention relates to a tissue culture and propagation method of stevia rebaudiana and a culture medium thereof, in particular to a tissue culture rapid propagation method and a culture medium thereof in which stevia leaves are used as explants to directly form buds. Background technique [0002] Asteraceae Stevia Stevin rebaudiann, Bertoni, hereinafter referred to as Stevia, is a perennial herb of the Asteraceae Stevia genus native to Paraguay, South America. Because the steviol glycosides contained in Stevia can control blood sugar, lower blood pressure, promote metabolism and It has the effects of treating diabetes, obesity, regulating gastric acid, and restoring nerve fatigue, and has stable physical and chemical properties, and has no fermentation characteristics. Its products have a relatively long shelf life, and are especially suitable as sweeteners for food for patients with diabetes, hypertension, etc. , health product additives, etc., hav...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 吴卫唐鑫宋伦杨昭李兴娇曾建威何晓洪陈艾萌
Owner SICHUAN AGRI UNIV
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