41 results about "Hairy root culture" patented technology

A method for the genetic transformation of transgenic Stevia rebaudiana mediated by Agrobacterium rhizogenes, in which the stem tip explants of Stevia rebaudiana after precultivation are immersed in the Agrobacterium rhizogenes bacterium liquid carrying the pRI 101-AN DNA plasmid of the gene of interest, After infection, co-cultivation and sterilization treatment, the hairy roots were cultured on the selection medium containing kanamycin, and induced the formation of kanamycin-resistant hairy roots on the explants, and induced the formation of hairy roots The callus differentiates into adventitious buds, and the adventitious buds continue to grow to form test-tube plantlets, and PCR amplification method is used to identify transgenic plants carrying the target gene. By adopting the technical scheme provided by the invention, the target gene can be introduced into Stevia rebaudiana, and compared with the genetic transformation of chemical methods such as polyethylene glycol, the link of isolation and cultivation from protoplasts is reduced; and the genetic transformation of physical methods such as gene guns In contrast, it does not require expensive instruments and equipment, and the operation technique is simple.

This invention concerns a genetic transformation method to induce hairy root culture of Morinda species, such as Morinda officinalis How, to increase production of therapeutically useful metabolites, related extracts of the roots, and methods to treat ulcerative colitis using extracts of Morinda species, such as Morinda officinalis How.

The invention provides an induced proliferation method for glehnia littoralis hairy roots. The method comprises the following steps: obtaining glehnia littoralis aseptic seedlings; culturing agrobacterium rhizogenes for infection; performing co-culture on the transformed explant; and inducing hairy root generation and hairy root culture proliferation. The formation of glehnia littoralis hairy roots is induced by utilizing the agrobacterium rhizogenes infection, an in-vitro hairy root culture system is established, the content of medicinal component coumarin is analyzed and measured, the content of medicinal component coumarin is contrasted with the content of coumarin in adventitious root and artificially cultured glehnia littoralis roots, the content of coumarin in the hairy roots is the highest, and the glehnia littoralis hairy roots has the advantages of fast growth, short period, relatively high yield of the secondary metabolite and the like and can be used for industrial production of glehnia littoralis coumarin; and moreover, a novel path is provided for industrial production of the secondary metabolite of the glehnia littoralis by utilizing the hairy roots in the future, the threatened plant is protected, and the ecological balance is maintained.

The invention belongs to the field of genetic engineering, and specifically relates to a method for producing chlorogenic acid and dicaffeoylquinic acid by using Agrobacterium rhizogenes to genetically transform stevia rebaudiana to establish a hairy root culture system. The content includes the preparation of stevia rebaudiana explants, the activation culture of Agrobacterium rhizogenes, the suspension culture and the production of chlorogenic acid and dicaffeoylquinic acid. The detection by PCR proved that the hairy root of Stevia rebaudiana was produced by transformation, and the detection by HPLC showed that the hairy root of Stevia rebaudiana could produce chlorogenic acid and dicaffeoylquinic acid. Since the hairy roots used have the characteristics of rapid growth on the hormone-free medium, the method has the advantages of simple operation, low cost, and not being restricted by natural conditions such as climate and land. It provides a stable and sustainable new drug source and food functional factor for the production of chlorogenic acid and dicaffeoylquinic acid by using hairy root culture. The large-scale production of acid provides a reliable source and material basis.

The invention provides a method for inducing the production of tripterygium wilfordii hairy roots by agrobacterium rhizogenes, which comprises the following steps: (1) obtaining aseptic explants; (2) preparing bacterial liquid of agrobacterium rhizogenes; (3) performing agrobacterium rhizogenes infection to induce hairy roots; (4) establishing a hairy root in-vitro culture system, wherein the establishment of the hairy root in-vitro culture system comprises the following steps: culturing hairy roots in a 1 / 2 MS solid medium containing 500 mg / L cefotaxime sodium under a dark culture condition with a temperature of 27 + / -1 DEG C, performing subculture once every 7 days, reducing the using concentration of cefotaxime sodium gradually during subculture till no bacterium is found, finally putting the completely aseptic hairy roots on the 1 / 2 MS solid medium for subculture and preservation. During the detection of the produced aseptic tripterygium wilfordii hairy roots, a considerable amount of triptolide is found to be contained in the hairy roots, and the content is significantly higher than the content of triptolide in traditional tripterygium wilfordii wild roots and tissue culture roots; the method has good prospects for industrial application. The production of triptolide by hairy root culture fills the gap of traditional Chinese medicine.

A process for culturing the hairy root of medusa saussura to obtain saussura polyose includes providing the aseptic test-tube plantlet of medusa saussura, infecting it with bacterial strain R1000, R1601, or LBA 9402, and suspending culture in the 1 / 2 MS culture medium. Its advantages are high content of polyose (5-10%), and high output rate of polyose (1-2 g / L).

The invention relates to a method for increasing the content of flavonoids in Scutellaria baicalensis, which belongs to the field of biotechnology; adopting biochemical engineering technology, using rare earth praseodymium to induce the hairy roots of Scutellaria baicalensis to increase the content of flavonoids, which specifically includes the following steps: (1) sticking Suspension culture of hairy roots of Scutellaria baicalensis: Weigh about 100 mg of fresh hairy roots of Scutellaria baicalensis, place them in a large-scale culture container, and then add 1 / 2 MS0 medium with a volume fraction of 80% for suspension culture; (2) Pr Treatment of elicitors on the hairy root culture of Scutellaria baicalensis: add 25mmol / L Pr3+ solution to the culture medium and place it at a constant temperature of 25°C±1.0°C for 7 days; (3) Extraction and detection of flavonoids ; The present invention uses rare earth praseodymium to improve the medium, which can efficiently produce a new culture system of baicalin flavones. Compared with the 1 / 2MS0 control medium, the production of flavones increases by 4.502 times; A new high-quality sustainable drug source.

The invention discloses a tissue culture method for inducing and proliferating bovine vigorous hairy roots. In the method, the cotyledons of the boulders are used as explants to induce hairy roots, and the method includes explant sterilization, Agrobacterium activation culture, hairy root induction culture, proliferation culture and the like. Advantages of the present invention: (1) adopt the cotyledons of Bovine vigor to transform as explants; (2) Establish the pre-culture time, bacterium concentration, infection time and co-cultivation time suitable for the induction of bovine vigor hairy roots; (3 ) determined the best bacteriostatic agent for explant bacteriostatic culture; (4) determined the best pre-cultivation, co-cultivation, bacteriostatic culture and subculture medium; (5) the whole set of method operation provided by the present invention It is simple, low in production cost, high in induction success rate, and fast in proliferation, and can sustainably obtain the hairy root of the plant, which can be used for large-scale cultivation of the hairy root of the plant to extract the main chemical components such as the polysaccharide of the plant, and has a good application prospect .