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Taxus chinensis var mairei coniferous endophytic fungus strain and application thereof

A southern yew and endophytic fungi technology is applied in the field of southern yew needle endophytic fungi, which can solve the problems of less research and achieve the effect of promoting biosynthesis and accumulation and great application value

Pending Publication Date: 2022-01-28
XUZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many research reports on the diversity, growth-promoting ability and paclitaxel production of endophytic fungi from Taxus chinensis, but there are few related studies on whether endophytic fungi can induce the accumulation of effective ingredients such as taxol from Taxus chinensis

Method used

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  • Taxus chinensis var mairei coniferous endophytic fungus strain and application thereof
  • Taxus chinensis var mairei coniferous endophytic fungus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation, cultivation, identification and preservation of Pseudodidymocyrtis lobariellae KL27:

[0030] Endophytic fungal strain KL27 was isolated by the following steps:

[0031] 1) The tissue material used for the isolation of strains was collected from the healthy wild southern yew in Lichuan Xingdoushan Nature Reserve (29°59′49″N, 109°0′6″E), and the fresh roots, stems and The coniferous leaf tissue wax was packaged in a ziplock bag at low temperature and brought back to the laboratory for storage at 4°C for subsequent isolation of plant endophytic fungi.

[0032] 2) Rinse the intercepted southern yew needles with running water for 2 hours, and ultrasonically clean the washed plant samples 3 times for 2-3 minutes each time, so as to prevent the difficult-to-wash substances attached to the surface of the plant tissue from affecting the separation results.

[0033] 3) Take out the southern yew needles and carry out disinfection treatment in an ultra-clean...

Embodiment 2

[0040] Embodiment 2: Southern yew endophytic fungus KL27 fermented liquid treats southern yew seedlings

[0041]1) Take the Pseudodidymocyrtis lobariellae KL27 obtained in Example 1, under aseptic conditions, pick the mycelium of an inoculation loop, inoculate it in 100mL sterile PDB liquid medium, place it at 28°C, and cultivate it on a shaker at 150rpm for 7 days. Filtrate under reduced pressure, and collect 90 mL of the filtrate.

[0042] 2) Pour the filtrate into a small watering can, and evenly spray it on the southern yew seedlings, and spray the same amount of distilled water on the southern yew seedlings in the control group. The coniferous samples were cut 7 days after treatment, dried and ground, and the target product was extracted for chromatographic detection.

Embodiment 3

[0043] Example 3: Effects of Southern Taxus endophytes on Paclitaxel Biosynthesis in Southern Taxus Needles

[0044] Dry the needles of the two kinds of yews obtained in Example 2 (the southern yew treated with the KL27 fermentation broth, and the southern yew as a control) at 45°C to constant weight and grind them into powder, and weigh 0.5-1.0 g into a 50mL centrifuge tube, add acidified methanol solution at a ratio of 1:20 (w / v), sonicate for 1 hour, filter with double-layer filter paper, and keep the filtrate. Transfer the filter residue to a 50mL centrifuge tube, add a corresponding volume of acidified methanol solution, sonicate for 1 h, filter with double-layer filter paper, keep the filtrate, and repeat this step once. The three filtrates were combined, evaporated to dryness under reduced pressure with a rotary evaporator at <40°C, and the acidified methanol solution was discharged to precipitate the detected substance. Dissolve with methanol and dilute to 1mL, sonica...

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Abstract

The invention provides a taxus chinensis var mairei coniferous endophytic fungus strain and application thereof, wherein the strain is Pseudodidymocyrtis lobariellae KL27 and is preserved in the China General Microbiological Culture Collection Center (CGMCC) on July 14, 2021, and the preservation number is CGMCC NO.22459. According to the strain Pseudodidymocyrtis lobariellae KL27, cultured fermentation broth is sprayed on taxus chinensis var mairei nursery stocks, so that biosynthesis and accumulation of paclitaxel in coniferous leaves of taxus chinensis var mairei can be effectively promoted, and the strain Pseudodidymocyrtis lobariellae KL27 has a relatively high application value.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a southern yew coniferous endophytic fungus and its application. Background technique [0002] Yew (Taxus sp.), also known as yew, is an evergreen tree or shrub belonging to the family Taxaceae. There are about 11 species and more than 200 cultivated species of Taxus in the world. There are 4 species, 1 variety and 1 hybrid in China, namely Northeast Taxus, Himalayan Taxus, Yunnan Taxus, Chinese Taxus, Varieties of southern yew and hybrid Mandia yew and so on. Taxus is a precious timber tree species with high horticultural ornamental value and high medicinal value. Paclitaxel, the anti-cancer active ingredient in yew, can effectively inhibit the proliferation and reproduction of cancer cells, and is a precious drug for treating cancer. But Taxus grows slowly, and the content of paclitaxel is extremely low. Therefore, it is of great significance to use modern biote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01G7/06A01G17/00C12R1/645
CPCC12N1/14A01G7/06A01G17/005
Inventor 蒋继宏曹小迎李露丹徐玲霞宛雯周降生
Owner XUZHOU NORMAL UNIVERSITY
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