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Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus and application thereof

A technology of endophytic fungi and coix, which is applied in the field of microorganisms, can solve the problems of limiting the growth cycle and quality of coix, scattered distribution, and high requirements for artificial cultivation techniques, and achieve great application value and the effect of promoting plant growth

Inactive Publication Date: 2016-06-01
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the growth of Coix is ​​mainly maintained by exogenous hormones, chemical fertilizers and soil nutrients, which largely limits the growth cycle and quality of Coix.
Although Coix coix resources have a vast area and a large amount of artificial cultivation has been carried out; however, their distribution is scattered, the resource renewal cycle is long, and the artificial cultivation technology requires high requirements, and the quality of medicinal materials varies.
With the discovery of many pharmacological activities of coix coix, its market demand is increasing, and the demand of the Chinese herbal medicine market cannot be met by directly harvesting coix coix natural and artificial resources
On the other hand, promoting the growth of Coix by fertilization and chemical reagents will also affect the quality of Coix and the clinical efficacy of Coix.

Method used

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  • Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus and application thereof
  • Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus and application thereof
  • Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The coix endophytic fungus of the present invention is a strain obtained from coix root cultivated in Taishun, Zhejiang. The coix endophytic fungus of the present invention is isolated and obtained according to the following steps: the collected fresh coix leaves, stems and seed kernels are washed with tap water, and after removing surface soil and dust, they are washed three times with deionized water. Blot the surface moisture with sterile filter paper, and carry out surface disinfection according to the following procedures: 75% ethanol, 30s→1% sodium hypochlorite, 5min→75% ethanol, 30s→sterile deionized water wash 3 times. After absorbing the residual moisture with sterile filter paper, use sterilized medical scissors or branch shears to cut the leaves, stems, and kernels into small tissue pieces (about 0.5cm×0.5cm for stems and leaves, and about 0.1cm×0.1cm for kernels). cm), 8 tissue blocks were randomly selected from each of the 3 parts, and each 4 tissue blocks ...

Embodiment 2

[0050] (1) Take the endophytic fungus of Coix coix, pick a small amount of hyphae with an inoculation needle under aseptic conditions, insert it into a sterilized solid PDA medium test tube, and activate and cultivate it at 25±1°C for 1-4 weeks. The morphology of the endophytic fungus of Coix (the 18th day) on the PDA medium is as follows: figure 1 shown;

[0051] (2) Mix wheat bran and cottonseed hulls evenly at a mass ratio of 1:1, add them into the prepared nutrient solution at a material-to-water ratio of 1:2, stir well, and autoclave at 121°C for 2 hours. The target bacterial species E.bromicola of the PDA culture dish is divided into small squares (1×1 cm) with an inoculation needle, inoculated on the surface of the solid bacterial culture medium, and cultivated for 2 weeks until the surface is covered with hyphae, and the solid bacterial species is obtained;

[0052] Such as figure 1 As shown, the coix endophytic fungus described in the present invention was cultured ...

Embodiment 3

[0054] (1) Soak coix seeds in 75% alcohol and shake for 3 minutes, transfer to 2.5% NaClO and soak for 15 minutes, repeat once, shake once every 5 minutes, rinse with sterile water 3-4 times, wash with sterile water Soak in water for 40 minutes, and finally rinse the seeds with sterile water 10 times. After sterilizing the Coix coix seeds, the seeds are placed on the MS medium without any hormones for aseptic culture to produce sterile seedlings (such as figure 2 -a). When the aseptic seedling grows to a height of 4-5cm (such as figure 2 -b), get the young leaves and roots of the aseptic seedlings and cut them into 2-3cm long cuts, and inoculate the cuts into callus induction medium (MS+3% sucrose+500mg / LCH (hydrolyzed casein)+2mg / L2, 4-D) for primary culture to induce callus formation (such as figure 2 -c). Then the callus is transferred to the culture medium for inducing clustered shoots (MS+0.1mg / LNAA+0.5mg / L6-BA) to induce clustered shoots (such as figure 2 -d), ...

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Abstract

The invention discloses a Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus which is obtained from a living plant body of the Poaceae coix plant Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf through the endophytic fungus separation and purification technology, and is identified through the microbial taxonomy to belong to the Epichloe bromicola.The bacterial strain is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO.11410.By co-culturing a Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus strain, Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf and Thale Cress, growth of Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf and Thale Cress is promoted, growth of main roots is restrained, development of lateral roots is promoted, plants can more easily derive nutrients and grow, the biomass of Thale Cress is improved, and the Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf endophytic fungus is an important microorganism for promoting growth of Coix lacroyma-jobi L.var.ma-yuen (Roman.) Stapf, Thale Cress and other gramineous plants.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to endophytic fungi of Job's tears and their application in promoting the growth of Arabidopsis or Job's tears. Background technique [0002] Coixlacryma-jobi L. var. mayuen. (Roman.) Stapf belongs to Gramineae (Gramineae), a plant of the genus Coix L., and is a species recorded in the Chinese Pharmacopoeia. The nutritional value is extremely high, and it has a variety of health effects. It is a resource of medicinal and edible crops that are loved and loved all over the world. Coix is ​​a commonly used traditional Chinese medicine, which has diuretic effects, invigorates the spleen, nourishes the lungs, dispels dampness and heat, eliminates edema, drains pus, and relieves diarrhea. Modern research proves that Coix has anti-tumor, immune-enhancing and anti-inflammatory effects. The research on Job's tears focuses on its chemical composition, pharmacological activity, extraction p...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P21/00C12R1/645
CPCA01N63/30C12N1/14C12N1/145C12R2001/645
Inventor 秦路平贾敏杨阳韩婷辛海量张巧艳李义敏孔周扬
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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