Technical flow applicable to mass production of polyploidy radix scutellariae excellent variety

A polyploid, skullcap technology, applied in application, plant regeneration, horticulture, etc., can solve the problems of difficult seed production and high production costs, and achieve the effects of improving quality and yield, reducing production costs, and creating a huge body size.

Inactive Publication Date: 2012-06-27
SINOBIOWAY BIO AGRI GRP CO LTD
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Problems solved by technology

[0004] The purpose of the present invention is to provide a complete technical process applicable to the large-scale production of tetraploid Scutellaria baicalensis, solve the problems of difficult seed production and high production costs in the large-scale production of tetraploid Scutellaria baicalensis, and obtain high-quality Scutellaria baicalensis Chinese medicinal materials

Method used

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  • Technical flow applicable to mass production of polyploidy radix scutellariae excellent variety
  • Technical flow applicable to mass production of polyploidy radix scutellariae excellent variety
  • Technical flow applicable to mass production of polyploidy radix scutellariae excellent variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 4

[0018] Induction of embodiment 1 tetraploid Scutellaria baicalensis

[0019] When the surface of the Scutellaria baicalensis callus differentiates into green buds, the callus is cut into small pieces with a diameter of 0.5 cm in an ultra-clean workbench, and the callus pieces are washed with 2% dimethyl sulfoxide (DMSO) Soak in 0.2% colchicine aqueous solution for 12-16 hours, rinse with sterile water for 3 times after treatment, inoculate into the culture medium for cultivation, make it differentiate into seedlings, the induction rate can reach 40.0%, the medium formula is as follows:

[0020] MS basic culture medium, sucrose 30g / L, agar 3.2g / L, 6-benzylaminopurine (6-BA) 0.5mg / L, chlormequat (PP 333 )0.5mg / L.

Embodiment 2 4

[0021] Example 2 Identification of tetraploid Scutellaria baicalensis-chromosomal microscopic examination

[0022] Each strain obtained by the treatment of Example 1 was induced to take root on the culture medium, and about 0.5 cm of the newly germinated test tube seedling root was cut, rinsed in distilled water, put into 0.1% colchicine solution at 20° C. for 30 min, and took out Afterwards, wash with distilled water for 3 times, put it into Carnot’s solution and fix it in the refrigerator for 2 hours, then wash with 95% ethanol, 70% ethanol, and distilled water successively for 3 times, isolate with 1M hydrochloric acid at 60°C for 5 minutes, and wash with distilled water for 3 times And hypotonic in distilled water for 30min. During microscopic examination, carefully cut out the root tip, put it in the center of the glass slide, add 1 drop of modified phenol-fuchsin staining solution, cover with a cover glass after 30 minutes, tap and press until the root tip is pressed int...

Embodiment 3 4

[0024] Example 3 Tetraploid Identification-Cell Flow Cytometry Analysis

[0025] Cut 1-2 leaves of Scutellaria baicalensis shoots, add 1ml of ice-cold buffer I to a petri dish, chop with a sharp blade, filter through a 40μm cell strainer, collect the filtrate, place it on ice, centrifuge at 500rpm for 5min, and stain the nuclei with PI staining solution. The DNA is fluorescently labeled, placed on ice, protected from light, and the plant ploidy is identified with a flow cytometer. After the sample is loaded, the flow cytometer automatically forms a DNA content distribution map representing the ploidy of the sample. The results of flow cytometry analysis of diploid Scutellaria baicalensis and tetraploid Scutellaria baicalensis cells are shown in the appendix figure 2

[0026] Buffer I: 45mmol / LMgCl 2 , 30mmol / L trisodium citrate, 20mmol / L MOPS, 0.1% TritonX-100, PH7.0.

[0027] PI staining solution: 50 μg / ml PI, 50 μg / ml RNase in buffer I.

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Abstract

The invention provides a technical flow applicable to the mass production of tetraploid radix scutellariae, which is characterized in that: radix scutellariae is used as a breeding material, a tetraploid radix scutellariae seedling with excellent and consistent characters can be rapidly obtained through the plant polyploidy breeding technology, the tissue culture and rapid propagation technology and the stem cuttage technology. The tetraploid radix scutellariae seedling belongs to the KT optimization, height of the tetraploid radix scutellariae seedling is 1.84 times of a reference, length of a leaf is 1.50 times of the reference, width of the leaf is 2.14 times of the reference, yield of a single plant is high, the yield of a single root is 1.70 times of a diploid reference, content of baicalin is improved by 1.86 percent compared to the reference, the fertility of the polyploidy radix scutellariea excellent variety is sterile, the entire growth period is 25 to 30 days longer than the reference, the tissue culture and rapid propagation technology is used for propagating the polyploidy radix scutellariea excellent variety to obtain original seedlings, and then the original seedlings are made into seedlings through the stem cuttage technology, the seedlings are transplanted to a field to be planted, about 5000 seedlings per Mu are planted, leaves and roots can be harvested in current year, about 40 to 50 kilograms of radix scutellariae tea can be produced per Mu, about 350 to 450 kilograms of roots can be produced, the content of the baicalin can reach 13.5 to 16.56 percent which is far higher than the requirement of pharmacopoeia that the content of the baicalin is not less than 8 percent, and the quality of the radix scutellariae is excellent.

Description

technical field [0001] The invention relates to a unique large-scale production process composed of technical links such as the creation of an excellent variety of tetraploid scutellaria baicalensis, rapid propagation of test-tube seedlings, cutting of original seedlings and field planting of seedlings, and belongs to the field of biotechnology breeding. Background technique [0002] Scutellariae baicalensis (Georgi) is one of the large varieties of Chinese medicinal materials unique to my country. It is mainly produced in North China, and Chengde, Hebei Province is the authentic production area of ​​Scutellaria baicalensis, with the best quality. Scutellaria baicalensis root can be used as medicine, the main active ingredient is baicalin, which has antibacterial, antiviral, antifungal, antipyretic, antihypertensive and anticancer effects. 70% of Chinese patent medicines contain Scutellaria baicalensis; Scutellaria baicalensis root extract, baicalein, and baicalin are used a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01G1/00
Inventor 刘敬梅谢莹周骅高春春
Owner SINOBIOWAY BIO AGRI GRP CO LTD
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