CN1038743A

CN1038743A - Mycen osmundicola and gastrodia seed germination culture technique

Info

Publication number: CN1038743A
Application number: CN 88103744
Authority: CN
Inventors: 徐锦堂; 冉砚珠; 郭顺星
Original assignee: Institute Of Medicinal Plant Resources Development Chinese Academy Of Medical Sciences
Current assignee: Institute Of Medicinal Plant Resources Development Chinese Academy Of Medical Sciences
Priority date: 1988-06-24
Filing date: 1988-06-24
Publication date: 1990-01-17

Abstract

A kind of osmund Xiao Ru and gastrodia seed germination culture technique, be under optimum conditions, with PDA medium culture one-level Mycen osmundicola bacterial classification → sawdust, the leaf of wheat bran second class inoculum → Fagaceae plant, sawdust, wheat bran three-class strain, select the blade of three-class strain then for use, place on the incubator foam-rubber cushion, keep saturation moisture, intensity of illumination 100-150Lux, under 25 ℃ of aeration conditions, can induce fruit body in 30 days.Broadcasting seeds of Gastrodia elata on the three-class strain blade, keep blade moistening, place under 25 ℃, seed can germinate after 20 days, evidence, and the leaf of the cultural hypha of separating with the Mycen osmundicola fruit body is mixed and is broadcast seeds of Gastrodia elata, germination rate is 19.65% after one month, and control group only is 8.98%, so turn out fruit body, is the key of bacterium classification and rejuvenation of spawn.

Description

Mycen osmundicola and gastrodia seed germination culture technique

The present invention relates to a kind of Mycen osmundicola (Mycena osmundicola Lange.) and rhizoma Gastrodiae (Gastrodiaelata Bl) seed symbiotic germination technology and Mycen osmundicola fruit body culture technique.

Rhizoma Gastrodiae is a kind of important rare traditional Chinese medicine, and the past, main dependence excavated wild hyoscine.Nearly two during the last ten years, domesticly utilizes the stem tuber vegetative propagation to cultivate successfully, but how degradation phenomena to occur for the amagaspecies fiber crops, and output descends significantly.Therefore, the research of planting seed sexual propagation has been carried out in some areas.Seeds of Gastrodia elata is minimum, no endosperm and other nutrient reserves, and seed sprouting is difficulty very.For a long time, people think that all the rhizoma Gastrodiae whole growth cycle (comprising the seed germination stage) all be unable to do without halimasch.Yet experimental results show that, halimasch not only can not provide seed germination needed nutrition, the effect of obvious suppression seed sprouting is arranged on the contrary, and study successfully " rhizoma Gastrodiae sexual propagation-leaf bacterium bed method ", this method be with planting seed on leaf, impel gastrodia seed germination with the nutrition that utilizes leaf.In recent years, from the protocorm of seed germination, isolate 12 kinds of seeds of Gastrodia elata symbiotic germination bacterial strains, cultivating these fungies with the leaf of Fagaceae plant mixes with the leaf that carries disease germs and broadcasts seeds of Gastrodia elata, germination rate can reach 2.83%-79.39%, the proof leaf is the medium of symbiotic germination bacterium, is the indirect source of nutrition of gastrodia seed germination.And seed germination is the symbiotic germination bacterium mycelia acquisition nutrition of invading embryo, utilizing leaf to cultivate out by digestion.The effect of symbiotic germination bacterium in the gastrodia seed germination process is undiscovered for a long time, and its classification position does not obtain scientific verification yet, makes the overall process of rhizoma Gastrodiae history of life be difficult to illustrate clear.Therefore, turn out the fruit body of seeds of Gastrodia elata symbiotic germination bacterium, determine that its classification section in Basidiomycetes belongs to, and design corresponding spawn culture and seeds of Gastrodia elata type of seeding, for research rhizoma Gastrodiae seeding technique, it is significant to improve rhizoma Gastrodiae output.Recently, induce fruit body from No. 01 better bacterium pearl of seeds of Gastrodia elata symbiotic germination bacterium, assay certificate is the Mycen osmundicola (Mycena osmundicola Lange) of Tricholomataceae (Trichomat-aceae) Mycena (Mycena).Do not see any report of relevant gastrodia seed germination and Mycen osmundicola relation through consulting the retrieval data at home and abroad, also do not see the report of artificial culture Mycen osmundicola entity and corresponding seeds of Gastrodia elata seeding technique.

The objective of the invention is to improve percentage of seedgermination and rhizoma Gastrodiae output, and the fruit body of turning out the symbiotic germination bacterium, identify its classification section and belong to, illustrate the rhizoma Gastrodiae overall process of the history of life theoretically.

Subject matter of the present invention: prove by experiment, seeds of Gastrodia elata must with the Mycen osmundicola symbiosis, obtaining nutrition by the digestion Mycen osmundicola could germinate, mix with the Fagaceae plant leaf that infects the Mycen osmundicola mycelia in the field and to broadcast seeds of Gastrodia elata, germination rate reaches 29.73% after two months, do not connecing (control group) on the leaf of bacterium and broadcast the same period, germination rate is 0%, and the successful seeds of Gastrodia elata seeding technique of corresponding research, prevented that the many kind fiber crops that cause for vegetative propagation of rhizoma Gastrodiae from degenerating, and opened whole secrets of the rhizoma Gastrodiae history of life.Studies show that, in the seeds of Gastrodia elata development stage, halimasch has the obvious suppression effect, and embryo is by digesting the Mycen osmundicola acquisition nutrition of invading and germinateing, the protocorm that germinates carries out vegetative propagation immediately, must set up symbiotic relation with halimasch again and just can finish its history of life.

Mycen osmundicola spawn culture technology:, glucose to boil water, agar) medium culture first class inoculum → sawdust, the leaf of wheat bran second class inoculum → Fagaceae plant, sawdust, wheat bran three-class strain with the PDA(potato.Under 25 ℃ of conditions, cultivate one and a half months, can produce three-class strain

Seeds of Gastrodia elata laboratory germination test technology: select three grades of blades of producing bacterial classification of Mycen osmundicola for use, being cut into diameter is 1.5-2.0 centimetre of size, broadcast seeds of Gastrodia elata, place on the incubator foam rubber cushion, every device 3-4 sheet splashes into distilled water, makes sponge keep saturation moisture content, place under 25 ℃ of conditions, broadcast the back and began to check germination rate in 20 days.The detection method of germination rate is, the wire of appropriate thickness is made the basket frame of 3 centimetres of diameters, middlely is divided into 3 centimetres lattice with superfine wire, is pressed on the blade seeds of Gastrodia elata number and germination protocorm number in the every little lattice of order statistics.The culture technique of Mycen osmundicola fruit body: select three grades of blades of producing bacterial classification of Mycen osmundicola for use, place on the incubator foam rubber cushion, often splash into distilled water, make sponge keep saturated water content, leaf water content is at 100-200%, be positioned under 25 ℃ of constant temperature, the environmental condition that maintains the circulation of air, scattered light intensity is 100-150Lux.Can induce the Mycen osmundicola fruit body about 30 days.The Mycen osmundicola mycelia repeatedly transfers, repeatedly also degradation phenomena can occur after the vegetative propagation, must turn out fruit body, by the bacterial classification of separate tissue or spore germination acquisition rejuvenation, mixes and broadcasts seeds of Gastrodia elata, and germination rate could obviously improve.Mix as the leaf of the cultural hypha of separating with fruit body and to broadcast seeds of Gastrodia elata, broadcasting back 30 days germination rates is 19.65%, and the same period in many generations vegetative bacterial classification mix and broadcast seeds of Gastrodia elata and compare, germination rate is 8.98%.So turn out fruit body, be the key of bacterium classification and rejuvenation of spawn.

Seeds of Gastrodia elata field sowing method: in 40 centimetres of dark caves of 30 centimetres of diameter, fill out earlier the wet leaf of one deck, three grades of production of seeds of Gastrodia elata and Mycen osmundicola bacterial classification is spread on the leaf after mixing and broadcasting, simultaneously three grades of halimasch kinds (bacterium branch) of cultivating in the culturing room are sprinkled in the cave, on straighten 5-10 centimetre the fresh wooden stick 3-5 root in footpath, cover thick 10 centimetres soil layer on the rod behind the lid layer leaf again.Adopt this method, in June, 1984, Beijing indoor seeding 16 caves, gather in the crops January next year, and 35.5 kilograms in rhizoma Gastrodiae is received in no hole altogether, and average cave is produced 2.2 kilograms, and the highest cave is produced and is reached 3.6 kilograms, and a year and a half gathered in the crops in November, 1985 after broadcasting, and on average the cave is produced 5.75 kilograms.Promote sowing 56500 caves in the Ningqiang County, Shaanxi Province, also obtain satisfied harvest.

Advantage of the present invention and good effect are: the leaf that utilizes Mycen osmundicola to cultivate is mixed and is broadcast seeds of Gastrodia elata, the field germination rate reaches 29.73%, broadcast rear a year and a half, cave output can reach 5.75 kilograms, and adopting rhizoma Gastrodiae generative propagation-leaf fungal bed method, the leaf of collecting from woodland is mixed and is broadcast seeds of Gastrodia elata, field germination rate only 7.43%, broadcast rear a year and a half, cave output only has 1.37 kilograms. Turn out the Mycen osmundicola fructification, not only identify the classification position of this bacterial classification, illustrated rhizoma Gastrodiae whole history of life, also make bacterial classification obtain rejuvenation, mix and broadcast behind the seeds of Gastrodia elata one month, germination percentage improves 10.67%.

Claims

1, the cultural method of a kind of Mycen osmundicola and seeds of Gastrodia elata symbiotic germination technology and Mycen osmundicola fruit body, it is characterized in that under suitable condition, go out Mycen osmundicola with specific medium culture and produce bacterial classification for three grades, it is mixed broadcast seeds of Gastrodia elata, can improve percentage of seedgermination and output.

2, according to the described technology of claim 1, it is characterized in that selecting for use three grades of blades of producing bacterial classification of Mycen osmundicola, broadcast seeds of Gastrodia elata, place incubator, keep 25 ℃ of temperature and saturated water content to cultivate.

3, according to the described technology of claim 2, the detection method that it is characterized in that germination rate, adopt the wire of appropriate thickness to make the basket frame of 3 centimetres of diameters, the middle lattice that is divided into 3 millimeters again with superfine wire, be pressed on the blade seeds of Gastrodia elata number in the order statistics lattice and germination protocorm number.

4, according to the described technology of claim 1, it is characterized in that the rhizoma Gastrodiae field sowing produces bacterial classification with three grades of Mycen osmundicolas and mix and broadcast seeds of Gastrodia elata, be sowed on the leaf of sowing cave, be sprinkled into three grades of halimasch kinds simultaneously.

5, in accordance with the method for claim 1, it is characterized in that to boil water with the PDA(potato, add glucose, agar) medium culture first class inoculum → sawdust, the leaf of wheat bran second class inoculum → Fagaceae plant, sawdust, wheat bran produce bacterial classification for three grades.

6, in accordance with the method for claim 5, it is characterized in that the condition of culture of fruit body is, 25 ℃ of optimum temperatures, leaf water content 100-200%, environment should maintain the circulation of air, the leaf of culture matrix and Fagaceae plant, scattered light illumination is 100-150Lux.

7, according to claim 5,6 described methods, it is characterized in that the fruit body of turning out by separate tissue or spore germination, obtain the bacterial classification of rejuvenation.