CN103026900B - Method for obtaining special mycorrhiza source for synthetizing forest establishment mycorrhizas - Google Patents

Abstract

The invention relates to a method for obtaining a special mycorrhiza source for synthetizing forest establishment mycorrhizas, namely a mycorrhiza source except artificially cultured mycelia and fungal spores, and belongs to the technical field of mycology under microbiology in biology. The method comprises the steps that aseptic forestal nursery stocks are cultured under an aseptic condition; during the period when fine roots grow out from the nursery stocks and the nursery stocks are less than one year old, the fine roots of the nursery stocks are artificially inoculated by adopting separated and purified mycelia or mycorrhizal fungi and are cultured for more than two weeks, so as to obtain the mycorrhizas; and the obtained mycorrhizas are cut into segments of length greater than or equal to 1mm by using sharp scissors or a diffusion knife, and are directly used as the mycorrhiza source for implementing nursery stock mycorrhization or forest establishment mycorrhization, or used for culturing the next generation of mycorrhiza inoculation sources. The technology for obtaining the inoculation sources, which is disclosed by the invention, is simple and is easy to operate; and the obtained mycorrhizas survive for a relatively long time, and the inoculation success rate is relatively high.

Description

The preparation method in the synthetic special bacterium of the mycorhiza that grows into forest source

Technical field

The present invention relates to a kind of preparation method of the synthetic special bacterium of the mycorhiza source of growing into forest, a kind of bacterium source except artificial cultured mycelia and fungal spore.Belong to the mycology technical field under microbiology in biology.

Background technology

Mycorhiza refers to the symbiont of fungi and plant roots.All fungies that can cause that plant forms mycorhiza are called mycorrhizal fungi.The feature of Applying Ectomycorrhizal Fungi is with mycelium, closely to surround the absorption root that Young Plant is tender to form fine and close sheath cover, and some sheath cover also grows mycelia or shoestring, has replaced the root hair of plant; The exoderm space between cells that part mycelia is invaded root forms special network structure, is called Hartig net; Mycelia does not generally enter among cortical cell.The fungi that forms ectotrophic mycorrhiza belongs to basidiomycetes more, as russule Zoopagales, Boletales and Tricholoma, also has a few species to belong to sac fungi, as Tuberale.Approximately have 3% plant to have ectotrophic mycorrhiza, wherein majority is arbor species, comprises angiosperm and gymnosperm.

Ectotrophic mycorrhiza can be strengthened the absorption of plant to mineral nutrient elements such as moisture and phosphorus, potassium, nitrogen, calcium, and to plant, provides the metabolites such as growth hormone, vitamin, the basic element of cell division, antibiotic and fatty acid, Promoting plant growth.Ectotrophic mycorrhiza can also improve plant to the resistance of pathogen infection and improve the resistance to extreme environments such as temperature, arid and peracid or mistake alkali or some heavy metals.In this syntaxial system, plant is for mycorrhizal fungi provides good ecotope and organic nutriment etc.Especially part alpine plant, as Pinaceae and Fagaceae, long-term and ectotrophic mycorrhiza symbiosis and producing compared with strong dependency, does not have ectotrophic mycorrhiza symbiosis cannot naturally to grow into forest.Many tests show to build up Va Mycorrhiza Seedling with the pure culture body inoculation sapling of Applying Ectomycorrhizal Fungi, when afforestation, play an important role.On dead soil, ectotrophic mycorrhiza improves forest survival rate and promotes that the effect of growing into forest is particularly evident.Therefore the Ministry of Forestry's the nineties in last century starts to widely popularize Mycorrhizal seedling afforestation technology, in the great Northwest desert virescence rebuild of Mycorrhizal Miao China, exhibits one's skill to the full.

The technology that makes Fine-root infectious bacteria root fungus and form symbiont is called Mycorrhizal technology.Mycorrhizal technology is not only exhibited one's skill to the full in above-mentioned Mycorrhizal seedling afforestation, and is the only way of the precious mycorhiza edible mushroom of development.The truffle mycorrhiza fungi technology that realizes at present industrialization is exactly an example application that economic benefit highlights.And edible VA Mycorrhizal Fungi leaved for development comprises cepe, matsutake, Russula vinosa, delicious lactarius, chicken fir, red beef liver, ox liver, wizened bacterium, but do not place restrictions on these kinds, it is world-renowned organic ticbit that quite a lot of kind is recognized.The Mycorrhizal of growing into forest technology is the prerequisite of these bacterium exploitations, and technology has a extensive future.

The edible VA Mycorrhizal Fungi that up to the present, can stably carry out scale, commercialization bionic cultivation only ferfas ( ttuber sp), this belong to honor ferfas ( tuber magnatumpora) for a plurality of kinds of representative can be synthetic by Seedling Inoculation, mycorhiza, garden style transplanting, standardized management obtain ferfas commodity, the history of commercialization cultivation is more century-old, technology comparative maturity.In recent years, red must false truffle ( rhizopogon rubescenstul.), pixie stool ( cantharellus cibariusfr.), delicious lactarius ( lactarius deliciosusgra) etc. (L:Fr.) imitate the technology path of ferfas bionic cultivation, also successively obtain the patent (Tan Zhuming, Fu Shaochun, Zhou little Ling, 2003) of bionic cultivation.Suillus lutens ( suillus lutus) bionic cultivation (Lv Mingliang, 2007) be after artificial infection in nursery forest bed direct fruiting.The common trait of the mycorhiza edible mushroom of the bionic cultivation that these successfully obtain is to realize Mycorrhizal seedling stage forest.

, comprise that China mainly carries out forestry nursery stock Mycorrhizal at present all over the world, the Mycorrhizal of growing into forest progress is slow.Trace it to its cause and comprise: one, the radicula children of forest Seedling Stage is tender, is easy to mycorhiza and invades, and shows, the easier Mycorrhizal of nursery stock in young age according to the people's such as Meng Fanrong (Meng Fanrong Shao Jing literary composition Qu Shaoyi, 1995) result of study; Its two, the soil in nursery is easy to manual control environment, until the miscellaneous bacteria of using disinfection soil to grow seedlings to reduce in mycorhiza inoculation is disturbed; Its three, the external condition such as seedling inoculation smooth warm water more easy to control is wet.Comparatively speaking, not only sylvan life inoculation is without above-mentioned advantage, must stand microbe species abundant in sylvan life soil, comprises the VA Mycorrhizal Fungi of surely growing in advance, all adverse effects to VA Mycorrhizal Fungi inoculation.Sylvan life inoculation difficulty is the bottleneck of ectotrophic mycorrhiza and the development of mycorhiza edible mushroom.Attempt to walk around this bottleneck, the way that adopts cepe symbiosis nursery stock to move into grow into forest is successfully (Hall I. R. Lyon A. J. E. Wang Y., 1998) also.

In Mycorrhizal technology, normally used inoculation source has artificial cultured mycelia and fungal spore, and artificial cultured mycelia time-to-live in the abundant forest soil of microbe species is very short, is difficult to inoculate successfully; Fungal spore can obtain longer survival period and rate of vaccination in forest soil, but because collection is difficult, cost is high, and cannot carry out artificial propagation, thus affect its use on producing, be mainly used at present the test of sylvan life Mycorrhizal.

Summary of the invention

The invention provides a kind of time-to-live long, the preparation method in the synthetic special bacterium of the mycorhiza source of growing into forest that success ratio of inoculation is higher.

The technical scheme that the present invention takes is as follows:

The preparation method in the synthetic special bacterium of the mycorhiza source of growing into forest of the present invention, the method is cultivated aseptic forestry nursery stock under aseptic condition, nursery stock, grow radicula to seedling growth between 1 anniversary, adopt the mycelium of separation and purification or the spore of VA Mycorrhizal Fungi to carry out artificial infection nursery stock radicula, cultivate more than 2 weeks, to obtain mycorhiza, the mycorhiza obtaining can be observed mycoclena and dense, the outward extending mycelia on radicula under the microscope, even can observe a large amount of shoestring (see figure 1)s.By the long segment of be cut into >=1mm of sharp scissors or cutting knife for mycorhiza, directly as bacterium source, implement nursery stock or the Mycorrhizal of growing into forest, or cultivate for mycorhiza inoculation source of future generation.

The present invention combines the advantage of existing two large class ectotrophic mycorrhiza inoculums, avoided drawback separately simultaneously, realize and be both easy to amount reproduction, can effectively improve again its time-to-live in soil and inoculation efficiency under natural forest, make the Mycorrhizal technology of the natural forestry hill of large area implement to become possibility.According to indoor measurement, at equal 15-20 ℃, in forest soil leachate, during 40hr, in the mycelium of the Russula fungi of artificial culture purified, there are a large amount of vacuoles, lose infected, in one week, all wither away.And the mycorhiza of the same bacterium that the present invention introduces can still keep more than 30% infected (seeing Fig. 1-3) after 4 weeks.

The present invention not only can obtain a large amount of bacterium sources that the time-to-live is long under natural forest, success ratio of inoculation is high, also makes up the breeding problem that minority is difficult to the VA Mycorrhizal Fungi of separation and purification simultaneously.

The inoculation source time-to-live provided by the invention can reach more than 4 weeks.(supplying the Russula fungi of examination under 15-20 ℃ of condition, the culture success ratio 30% after in vitro 4 weeks).

Remarkable advantage of the present invention: 1, technology is simple: nursery stock Mycorrhizal is a very ripe technology, therefore that the present invention obtains inoculation source technology is simple, easy operating; 2, effect is remarkable: key is that the mycorhiza easily obtaining is cut into after small pieces, it infects in the temperature of relation, forest root system under wet condition and soil and can maintain vigour to more than 1 week in suitable foundation, guarantees that mycelia on mycorhiza or shoestring have time enough to extend and succeed to infect; 3, for the VA Mycorrhizal Fungi that is difficult to separation and purification, also can as initial inoculation source, obtain amount reproduction by its spore, significantly expand inoculation source quantity, on producing, there is very important meaning.

Accompanying drawing explanation

Fig. 1 is the entity photo of observing the nursery stock mycorhiza of a large amount of shoestrings.

Fig. 2 is mycorhiza (left figure) (normally) and mycelium (right figure) (normally) microphoto after processing 1hr.

Fig. 3 is mycorhiza (left figure) (normally) and mycelium (right figure) (occurring a large amount of vacuoles in mycelium) microphoto after processing 40hr.

Fig. 4 processes mycorhiza (left figure) (a large amount of miscellaneous bacterias appear in mycorhiza periphery, but mycorhiza still has certain vigor) and mycelium (right figure) (mycelium dissolving) microphoto after 1 week.

Embodiment

1, the present invention is applied to Russula fungi bionic cultivation, from the white pin russule of Putian ( russula sp) isolate pure mycelia, after expand cultivating, be inoculated in triangular flask on the aseptic culture red paper mulberry seedling radicula of 20 days, after 1 month, transplant to aseptic red soil, cultivate again after 5 months, shoestring (white pin russule and red paper mulberry symbiosis) mycorhiza is covered with in intercepting, be cut into the root segment of 3mm size, with take the inoculation additive that fine sand and peat soil be main component and mix thoroughly, in Di Kou township, Jianou City, select the original hill without this bacterium, adopt sylvan life inoculation technique to be seeded on red paper mulberry radicula, successfully after 2 years, obtain fruit body, cave success ratio of inoculation 32%.The report of expert's site inspection shows, in the region that has the inoculation of listing, observe the fruit body of Russula fungi, each fruiting point only can be observed the fruit body of 1 or 2 Russula fungi, and the wild russule observing in the wild fruiting region of locality, each fruiting point can be observed a plurality of fruit bodys and be all living creatures's situation.The Russula fungi form of the sporophore shape of this inoculation fruiting and local periphery hill has notable difference, and performance is that strange land russule inoculates successfully.

2, the present invention is applied to " cepe bionic cultivation ", collect plan cepe from Yunnan ( boletus sp), adopt spore inoculating on aseptic red paper mulberry seedling, after 3 months, collect mycorhiza and inoculate, expand fast inoculation source, adopt afterwards inoculation technique as above, be seeded on the red paper mulberry hill that there is no the Putian of this bacterium Chang Taixiang, the Second Year of sylvan life inoculation, in inoculation, district collects a large amount of plan cepes.

3, the present invention is applied to yield increasing fungus (VA Mycorrhizal Fungi) inoculation of " Russula vinosa protection ground fungal diversity yield-increasing technology ", makes the successfully main russule vitellarium popularization in Fujian Province of this achievement.

list of references

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2、Hall?I.?R.?Lyon?A.?J.?E.?Wang?Y.?and?Sinclair?L.?Ectomycorrhixal?fungi?with?edible?fruiting?Bodies?2.? Boletus?edulis，?Economic?Botany，?1998，?52(1):44-56.

3, Lv Mingliang, bolete artificial cultivation is succeeded, Beijing Agriculture practical technique, 2007 (6):

4, Meng Fanrong Shao Jing literary composition Qu Shaoyi, Shao Gui is loyal etc. for a long time along Wu Qing virtue grandson, affects the Primary Study Journal of northeast Forestry university of Original Artificial Inoculation of Ectomycorrhizal Fungi, 1995,23V, 6:8-13

5, Liang Yu, Guo Liangdong, mark put down, the effect of mycorrhizal fungi in the ecosystem, Acta Phytoecologica Sinica, 2002,26 (6): 739～745

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Claims (1)

1. the method in the synthetic special bacterium of the mycorhiza that grows into forest source, it is characterized in that: the preparation method in this bacterium source for cultivating aseptic forestry nursery stock under aseptic condition, nursery stock, grow radicula to seedling growth between 1 anniversary, adopt the mycelium of separation and purification or the spore of VA Mycorrhizal Fungi to carry out artificial infection nursery stock radicula, cultivate more than 2 weeks, to obtain mycorhiza; By the long segment of be cut into >=1mm of sharp scissors or cutting knife for the mycorhiza of acquisition, directly as bacterium source, implement to grow into forest Mycorrhizal or nursery stock Mycorrhizal, or cultivate for mycorhiza inoculation source of future generation.