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Suspension culture method for pinellia ternata artificial embryo synchronization

A suspension culture and synchronization technology, applied in horticultural methods, botanical equipment and methods, horticultural tools/equipment, etc. The effect of quality purification and rejuvenation, solving dosage and serious disease

Active Publication Date: 2015-12-02
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although artificial planting has alleviated the contradiction between supply and demand of pinellia medicinal materials to a certain extent, due to factors such as the large amount of seeds per mu, frequent continuous cropping diseases, and serious germplasm degradation, most farmers are discouraged from planting pinellia.
Although the success of Pinellia tissue culture technology has provided a new way to solve Pinellia reproduction and germplasm innovation, but the planting workload of tissue culture seedlings is heavy, the management procedures are cumbersome, and the survival rate is low, so this technology is widely used in the large-scale cultivation of Pinellia. of little practical significance

Method used

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  • Suspension culture method for pinellia ternata artificial embryo synchronization
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  • Suspension culture method for pinellia ternata artificial embryo synchronization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: callus induction and subculture

[0024] The material pinellia [ Pinellia ternata (Thunb.) Breit.] collected from Hezhang County, Guizhou.

[0025] Select the leaves or petioles of Pinellia ternata with good growth as explants, use MS as the basic medium, and use explants, 2,4-D, 6-BA three-factor three-level orthogonal experiment design to test the callus Induced culture. Culture conditions: temperature (25±1) ℃, light intensity 2000lx, light time 12h / d. The callus induction rate and its fresh weight were counted (Table 1).

[0026] It can be seen from Table 1 that different treatment combinations had little effect on the callus induction rate of Pinellia ternata, but had a great influence on the amount of callus formed, the color and texture of callus. Conclusion: Inoculate leaves in MS+2,4-D1.0mg / L+6-BA1.5mg / L or petioles in MS+2,4-D1.0mg / L+6-BA1.0mg / L medium Above (the pH of the medium is 5.8, the same below), the induced callus grows well and loo...

Embodiment 2

[0030] Example 2: Construction of Microcell Aggregate Suspension System

[0031] Loose calli with more than 3 subcultures were selected and inoculated in liquid MS medium supplemented with different types and concentrations of plant growth regulator substances and different concentrations of sucrose and CH. Four factors, 2,4-D, 6-BA, sucrose, and CH were used in a three-level orthogonal experiment, and the rotation speed of the shaker was 90 r / min, and other culture conditions were the same as in Example 1. By examining the effects of different treatments on the proliferation of microcell clusters, it was concluded that the microcell cluster suspension medium was mixed with MS+2,4-D1.0mg / L+6-BA0.5mg / L+CH300mg / L+sucrose 40g / L The effect is better (Table 3). When cultured in this medium, the microcell mass proliferates rapidly and has a high degree of dispersion, and the culture is milky white to light yellow.

[0032]

Embodiment 3

[0033] Embodiment 3 Artificial Seed Embryo Suspension Culture

[0034] Take the cell mass that has been cultured in suspension for more than 3 generations, use MS+6-BA0.5mg / L+CH300mg / L+sucrose 30g / L as the basal medium, add different types and concentrations of auxin substances, and shake the table at a speed of 75r / min , and all the other culture conditions are the same as in Example 1. By examining the effects of different composition combinations on the expansion of cell clusters, the conclusion is that the addition of IBA1.0mg / L in the basal medium has a better effect on the expansion of cell clusters (Table 4). In this medium, the expansion speed of cell clusters is fast and uniform. The degree of differentiation is high, the texture is compact, and the cell mass is white or milky white.

[0035]

[0036] The expanded cell mass was inoculated in MS+CH300mg / L+sucrose 10g / L as the basal medium, and different concentrations of 6-BA and IBA were added for suspension diffe...

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Abstract

The invention discloses a suspension culture method for pinellia ternata artificial embryo synchronization. Firstly, a pinellia ternata aseptic seedling blade or petiole is taken as an explant to induce and generate a callus, then a loose callus suitable for suspension culture is obtained by adjusting subculture media and subculture times, a pinellia ternata fine and uniform cell cluster suspension system is built through suspension culture, and finally, an optimized pinellia ternata artificial embryo synchronization culture condition is constructed by adjusting the plant growth adjustment material types and concentration, sucrose concentration, casein hydrolysate (CH) concentration and illumination intensity. According to the suspension culture method for pinellia ternata artificial embryo synchronization, a pinellia ternata artificial embryo is obtained by suspension culture, the suspension culture method has the advantages that the multiplication rate is fast, the synchronization degree is high, the detoxication effect is good, the operation is easy and convenient and the like, and a novel effective way is proved for construction of the pinellia ternata seed and seedling industrialization technology.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for inducing loose callus of Pinellia pinellia, subculture proliferation and differentiation culture of Pinellia pinellia artificial seed embryo. Background technique [0002] pinellia[ Pinellia ternata (Thunb.) Breit.], also known as Mayu fruit, three step jump, three leaves, swallow tail, etc., is a perennial herbal medicinal plant of the genus Pinellia in the family Araceae, widely distributed in my country (except Inner Mongolia, Heilongjiang , Jilin, Qinghai, Xinjiang, Tibet, other provinces are distributed). Pinellia is a bulk medicinal material. Its tubers are used as medicine. It tastes pungent, warm in nature, and poisonous. Pinellia is contained in more than 200 kinds of Chinese patent medicines. [0003] With the in-depth research on pinellia, its medicinal value is gradually reflected, and the amount of medicinal use is increasing y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张明生刘贵贤齐政吕享高晓峰王汪中韦红边
Owner GUIZHOU UNIV
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