CN101953261B - Application of jewel orchid endophytic fungi to jewel orchid seedlings - Google Patents
Application of jewel orchid endophytic fungi to jewel orchid seedlings Download PDFInfo
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- CN101953261B CN101953261B CN2009102100732A CN200910210073A CN101953261B CN 101953261 B CN101953261 B CN 101953261B CN 2009102100732 A CN2009102100732 A CN 2009102100732A CN 200910210073 A CN200910210073 A CN 200910210073A CN 101953261 B CN101953261 B CN 101953261B
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Abstract
The invention relates to application of endophytic fungi, in particular to the application of symbiotic cultivation of jewel orchid endophytic fungi and jewel orchid tissue culturing seedlings. The endophytic fungi are Rhizoctonia spp. and are separated from the root part of the wild jewel orchid by a tissue separation method. The Rhizoctonia spp. is collected in China General Microbiological Culture Collection Center (CGMCC for short), and the address is Institute of Microbiology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing. The Rhizoctonia spp. infects the jewel orchid tissue culturing seedlings to form symbiotic seedlings, so that the application can improve stress resistance of the jewel orchid tissue culturing seedlings, improve survival rate and shorten growth cycle. Therefore, the production cost is reduced and industrialized production is realized.
Description
Technical field:
The present invention relates to a kind of application of endogenetic fungus, specially refer to a kind of Herba Anoectochili roxburghii endogenetic fungus and cultivate the application in the seedling in the Herba Anoectochili roxburghii group.
Background technology:
Herba Anoectochili roxburghii belongs to the per nnial herb that the orchid family is opened the lip Cymbidium, is the wider a kind of traditional valuable ingredient of traditional Chinese medicine of use among the people, is used for treating pulmonary tuberculosis, cough due to lung-heat, rheumatic arthritis etc. more.In recent years, Herba Anoectochili roxburghii was used to treat the effects that difficult and complicated illness obtained such as hypertension, mellitus and tumour, caused the attention of the world of medicine day by day, and especially in Taiwan, Herba Anoectochili roxburghii has the title of " king of Chinese medicine ".
Yet Herba Anoectochili roxburghii is very rare in the nature resource, and wherein reason is very harsh and receive nature animalcule and people's the destruction to the requirement of ecotope except it, and main is that the seed of Herba Anoectochili roxburghii is very little, does not have complete its growth of embryo confession.Under field conditions (factors), need infecting for it provides nutrition of endogenetic fungus just can develop into seedling, this also is universal characteristics of orchid.Endogenetic fungus provides the nutritive substance except can be orchid; Can also promote its absorption to mineral substance; Produce required VITAMINs of growth and development of plants and plant hormone substance, enhancement of plant resistance and disease resistance (being published in " Shanghai Agricultural journal " the 2005.21st (3) phase " function of orchid endogenetic fungus and application prospect [J] ") with reference to Zhou Deping etc.This shows, adopt Herba Anoectochili roxburghii tissue cultured seedling and the symbiotic method of mycorrhizal fungi, advantages such as can promoting plant-growth, enhancement of plant resistance and disease resistance is arranged, can overcome present tissue cultured seedling poor growth, weak, the more high shortcomings of mortality ratio of resistance.At present; Aspect anoectochilus symbiosis; Do not report as yet that abroad domestic Guo adopts the little mushroom of the stem of noble dendrobium (Mycenadendrobii), Mycen osmundicola (Mycena osmundicola) along magnitude; Mycena orchidicola (Mycenaorchidicola), Mycena anoectochila) (Mycena anoectochila) and anoectochilus symbiosis (number of patent application: 02100779.9; The patent No.: ZL 200510117703.3), effect unsatisfactory (high slightly, Guo is along magnitude " middle medicinal herbs " the 2002.33rd (6) interim " three kinds of endogenetic fungus are to the research of Herba Dendrobii, Herba Anoectochili roxburghii growth effect ").
Summary of the invention:
The object of the present invention is to provide a kind ofly can form symbiotic relationship and promote it to grow, improve its breeding potential, survival rate, and strengthen the application of the Herba Anoectochili roxburghii endogenetic fungus of its resistance with Herba Anoectochili roxburghii.
The application of a kind of Herba Anoectochili roxburghii endogenetic fungus in Herba Anoectochili roxburghii is grown seedlings, this endogenetic fungus are rhizoctonia (Rhizoctonia spp.).
Described rhizoctonia (Rhizoctonia spp.) is rhizoctonia (Rhizoctonia sp.) JSH-1 or GXSH-5 or GXSH-6; These three bacterial strains all have been preserved in China Microbial Culture Preservation Commission common micro-organisms center; The address is a Datun Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica; Its deposit number and date are respectively JSH-1:CGMCCNO.3074, preservation date on May 20th, 2009; GXSH-5:CGMCC NO.3273, preservation date on September 7th, 2009; GXSH-6:CGMCC NO.3274, preservation date on September 7th, 2009.
Said rhizoctonia (Rhizoctonia sp.) JSH-1 adopts tissue isolation from the separating obtained mycorrhizal fungi of wild Herba Anoectochili roxburghii root.
Wherein, said rhizoctonia (Rhizoctonia sp.) JSH-1 solid culture is characterized as (Fig. 1, Fig. 2): 28 ℃ of cultivations on potato dextrose agar (PDA) substratum, mycelia initial stage milk yellow; The later stage light tan is the homogeneous radiation shape to outgrowth, and growth velocity is 2.5～3.0cm/d; Aerial hyphae is more; Form the donut that does not wait at interval, bacterium colony begins to occur oily tawny sclerotium after 6 days, is distributed in the donut place more.
Said rhizoctonia (Rhizoctonia sp.) JSH-1 morphological specificity is (Fig. 3):
1). do not form any spore in asexual generation, the mycelia branch is hung at its origin place and is contracted, and barrier film is arranged, no clamp connexion; Rectangular or the acute angle of mycelia branch has sclerotium, and hyphal diameter is 4～7 μ m; Most cells have two nucleus, and individual cells nuclear is arranged individually, and nucleus is rounded.
2). sexual generation, do not find.
Other two strain bacterial strain rhizoctonia (Rhizoctonia sp.) GXSH-5 of the present invention and rhizoctonia (Rhizoctonia sp.) GXSH-6 and rhizoctonia (Rhizoctonia sp.) JSH-1 have following common trait: asexual generation, do not form any spore, and the mycelia branch is hung at its origin place and is contracted; Barrier film is arranged; No clamp connexion, the rectangular or acute angle of mycelia branch, diameter is 4～7 μ m; Most cells have 3 or above nucleus, and sclerotium is arranged.
Anoectochilus symbiosis seedling is cultivated in said rhizoctonia (Rhizoctonia sp.) JSH-1 or GXSH-5 or GXSH-6 and the symbiosis of Herba Anoectochili roxburghii tissue cultured seedling.
The mode that said symbiosis is cultivated is with the whole strain of Herba Anoectochili roxburghii group training seedling and said rhizoctonia (Rhizoctonia sp.) JSH-1 or GXSH-5 or GXSH-6 co-cultivation.
Above-mentioned co-cultivation is in symbiotic culture medium, under the aseptic condition, to carry out; Perhaps also can be in cultivation medium, carry out under the open condition.
When rhizoctonia of the present invention (Rhizoctonia spp.) is used, cultivate rhizoctonia (Rhizoctonia spp.) earlier in Herba Anoectochili roxburghii group cultivation seedling, cultivate with the mode and the symbiosis of Herba Anoectochili roxburghii group training seedling of mycelia agar block or microbial inoculum then, concrete operations are following:
1). the making of mycelia agar block and microbial inoculum
Mycelia agar block: adopt the PDA substratum; (Rhizoctoniaspp.) is inoculated on the culture medium flat plate with the rhizoctonia after the activation; 28 ℃ of dark culturing 2～4 days, (diameter is 0.5～1cm) or with the mycelia agar block of the inoculating needle of calcination block hitting or the picking length of side 0.4～0.7cm on colony edge with sterilized punch tool.
Microbial inoculum: rice bran or wheat bran and water are prepared burden according to (1: 1)～(2: 1) weight ratio; Regulate water cut to 55～60% (possess globule appearance but do not flow down) with PDB, put into the white disk that is used for sterilizing, four layers of gauze of surface coverage with hand-tight; 121 ℃ of sterilization 1h; Rhizoctonia (Rhizoctonia spp.) the mycelia agar block of length and width 0.3～0.8cm is inserted in the cooling back, cultivates 6～10 days for dark 28 ℃, rubs with the hands broken before using.
2). rhizoctonia (Rhizoctonia spp.) and anoectochilus symbiosis type of rearing
Mode a: receive whole strain of aseptic Herba Anoectochili roxburghii group training seedling and rhizoctonia (Rhizoctonia spp.) mycelia agar block on the symbiotic culture medium simultaneously; Each bottle inserts three strain groups training seedling; Under the pure culture condition, cultivated 2～3 months for 20～25 ℃; Intensity of illumination is 800～3000lx, 12 hours every days.
Mode b: be 1～5% microbial inoculum and matrix mixing with weight ratio; Place plansifter, flowerpot or other some hard holder basins, then plant and refined 15～60 days Herba Anoectochili roxburghii group of seedling training seedling, every strain spacing is 4～7cm; Intensity of illumination is 500～4000lx; Relative humidity is 75%～95%, and temperature is 20～30 ℃, cultivates and gathers after 5～7 months.
Adopt rhizoctonia according to the invention (Rhizoctonia spp.) to infect the Herba Anoectochili roxburghii tissue cultured seedling and form symbiosis seedling, can make the Herba Anoectochili roxburghii tissue cultured seedling strengthen resistance, improve survival rate, shorten growth cycle, thereby reduce its production cost, realize industrialization production.
Description of drawings:
Fig. 1 is that rhizoctonia (Rhizoctonia sp.) JSH-1 cultivates 2 days colonial morphology on the PDA substratum;
Fig. 2 is the sclerotium form that rhizoctonia (Rhizoctonia sp.) JSH-1 cultivated on the PDA substratum 7 days;
Fig. 3 is rhizoctonia (Rhizoctonia sp.) JSH-1 individual morphology and safranin O-KOH nucleus dyeing, 400 *;
State when Fig. 4 rigidly connects kind for Herba Anoectochili roxburghii;
Fig. 5 is the Herba Anoectochili roxburghii contrast seedling growth conditions of having grown 60 days;
Fig. 6 is the anoectochilus symbiosis seedling growth conditions of having grown 60 days;
The hyphal knot that Fig. 7 has withered away in the cortex parenchyma cell for field planting, 100 *;
The hyphal knot that Fig. 8 does not wither away in the cortex parenchyma cell for field planting, 400 *.
Embodiment:
Below in conjunction with specific embodiment and accompanying drawing the present invention is further specified.
Embodiment one:
Earlier behind the slant tube actication of culture of cryopreservation, change rhizoctonia (Rhizoctonia sp.) JSH-1 over to the PDA flat board, 28 ℃ of dark culturing two days are subsequent use.
(diameter is 0.5～1cm) or with the mycelia agar block of the inoculating needle of calcination block hitting or the picking length of side 0.4～0.7cm on colony edge with sterilized punch tool.
Choosing highly is the more sturdy Fujian Herba Anoectochili roxburghii group training seedling of 5 ± 0.5cm, receives on the symbiotic culture medium with whole strain mode, and every bottle graft is gone into 3 young plants, and picking rhizoctonia (Rhizoctonia sp.) JSH-1 mycelia agar block places between the 3 strain tissue cultured seedling; Above-mentioned all operations process is all operated on aseptic super clean bench.
Culture condition: temperature: 25 ℃; Intensity of illumination: 1500lx, light application time: 12h/d; Incubation time: 80 days.
Interpretation of result:
1. the Herba Anoectochili roxburghii tissue cultured seedling connects upgrowth situation behind the bacterium
Cultivate the initial stage altogether at tissue cultured seedling and rhizoctonia (Rhizoctonia sp.) JSH-1, the leaf look turns to be yellow slightly, and plant strain growth is not obvious, and about 20 days, plant base portion stipes begins to occur protocorm.Afterwards, plant strain growth is rapid, and stem is chap gradually, and blade is dark green gradually, and aerial root is more and more flourishing, have in addition be inserted in the substratum.And control group, plant strain growth is thin and weak, and here blade some roll up, never the appearance (Fig. 4～6) of protocorm.
2. connect the variation of Herba Anoectochili roxburghii tissue cultured seedling increment (fresh weight and plant height) behind the bacterium
Carry out the mensuration of increment and adopt tissue isolation heavily to separate former bacterial strain with the contrast seedling having cultivated trimestral anoectochilus symbiosis seedling: average plant height before average plant height rate of increase (%)=(average plant height before the average plant height in processing back-processing) * 100/ processing; Average fresh weight before average fresh weight rate of increase (%)=handle (handling the preceding average fresh weight of the average fresh weight in back-processing) * 100/.The result is as shown in table 1:
Table 1. bacterial strain JSH-1 is to the influence of Fujian Herba Anoectochili roxburghii group training growth of seedling situation
(annotate: data are three repetition MVs, adopt the Tukey method in the DPS statistical software, are illustrated in significant difference on 0.01 level between different lowercase marks.)
Embodiment two:
At first make microbial inoculum, optimum making method is: rice bran or wheat bran and water are prepared burden according to 1: 1 weight ratio, regulate water cut to 50%～60% (possess globule appearance but do not flow down with hand-tight) with PDB; Put into sterilized white disk; Four layers of gauze of surface coverage, 121 ℃ of sterilization 1h, rhizoctonia (Rhizoctonia sp.) the JSH-1 mycelia agar block of length and width 0.5cm is inserted in the cooling back; Cultivated 8 days for dark 28 ℃, rub with the hands broken before using.
According to 3% weight ratio and culture substrate uniform mixing, culture substrate is the mixture of thin peat and perlite (3: 1) with the microbial inoculum of making.The culture substrate that then will be mixed with microbial inoculum places plansifter, and thickness is 3cm, has refined 30 days Herba Anoectochili roxburghii tissue cultured seedling of seedling on the kind, and every strain spacing is 5cm, and intensity of illumination is controlled between 800～3500lx, and relative humidity is 85～90%, and temperature is 22～28 ℃.
Interpretation of result: plant and just observe the appearance that there is protocorm in symbiosis seedling stem after 5 days, protocorm growth rate is up to 93.79% after 27 days.Symbiosis seedling growth during this time is rapid, and new after two months long blade is 2～3, and average maximum width of blade is 1.25cm, and the illumination poor growth, new long blade is 1～2, average maximum width of blade is 0.85cm.Randomly draw a few strain symbiosis seedlings its protocorm is carried out the free-hand section observation, in its cross section cortex parenchyma cell, all find a large amount of hyphal knots, show the successfully foundation (Fig. 7, Fig. 8) of Herba Anoectochili roxburghii-endogenetic fungus syntaxial system.
Embodiment three, embodiment four all make an experiment, analyze according to embodiment one, embodiment two said Step By Conditions respectively with rhizoctonia (Rhizoctonia sp.) GXSH-5;
Embodiment five, embodiment six all make an experiment, analyze according to embodiment one, embodiment two said Step By Conditions respectively with rhizoctonia (Rhizoctonia sp.) GXSH-6;
Above-mentioned experimental result proves that this two bacterial strain GXSH-5, GXSH-6 can infect the Herba Anoectochili roxburghii tissue cultured seedling equally and form symbiosis seedling, and tangible growth promoting function is arranged.
In sum and according to teachings of the present invention; Adopt rhizoctonia according to the invention (Rhizoctonia sp.) and other rhizoctonias to infect the Herba Anoectochili roxburghii tissue cultured seedling and all can form the mycorhiza symbiosis seedling; All can make the Herba Anoectochili roxburghii tissue cultured seedling strengthen resistance, improve survival rate, shorten growth cycle; Thereby reduce its production cost, realize industrialization production.
The rDNA-ITS base sequence of rhizoctonia according to the invention (Rhizoctonia sp.) JSH-1 is following:
1 tccgtaggtg?aacctgcgga?aggatcatta?ttgaatttta?atgaggagtt?ggttgttgct
61 ggccttggga?tttttttaac?taattattcc?aggcacttgt?gcacaccttc?tctttcatcc
121?acacacacct?gtgcacttgt?gagacagact?gtggggactt?tattgaaccc?cgtctgtcta
181?cttaatttac?acacactcta?tttaatttaa?actgaatgta?attgatgtaa?cgcatctaat
241?actaagtttc?aacaacggat?ctcttggctc?tcgcatcgat?gaagaacgca?gcgaaatgcg
301?ataagtaatg?tgaattgcag?aattcagtga?atcatcgaat?ctttgaacgc?accttgcgct
361?ccttggtatt?ccttggagca?tgcctgtttg?agtatcatga?aatcttcaaa?gtaaaccttt
421?tgttaattca?atgggttctt?tactttggta?ttggaggtct?ttgcagtctc?acaactgctc
481?ctctttgttc?attagctgga?tctcagtgtt?atgcttggtt?ccactcggcg?tgataagtat
541?ctatcgctga?ggacacctgt?aaaaaggtgg?ccaaggtaaa?tgcagatgaa?ccgcttctaa
601?cagtccattg?acttggacaa?ataaatattt?atgatctgat?ctcaaatcag?gtaggactac
661?ccgctgaact?taagcatatc?aataagcgga?gga
The rDNA-ITS base sequence of said rhizoctonia (Rhizoctonia sp.) GXSH-5 is following:
1 tccgtaggtg?aacctgcgga?aggatcatta?atgaattaat?gtggagttgg?ttgtagctgg
61 tccgctggat?tttttaaaat?aaattccttt?ggacatgtgc?acgccttctc?tttcatccac
121?acacacctgt?gcacctgtga?ggcagttgtt?ggggacttta?ttgtaccctc?tctgtctact
181?taatttacac?aaactcaatt?tattttaaaa?cgaatgtaat?tgatgtaacg?catctaatac
241?taagtttcaa?caacggatct?cttggctctc?gcatcgatga?agaacgcagc?gaaatgcgat
301?aagtaatgtg?aattgcagaa?ttcagtgaat?catcgaatct?ttgaacgcac?cttgcgctcc
361?ttggtattcc?ttggagcatg?cctgtttgag?tatcatgaaa?tcttcaaagt?caatcttttt
421?gttaattcaa?ttggttgtgc?tttggtattg?gaggtctttt?gcagcttcac?acctgctcct
481?ctttgttcat?tagctggatc?tcagtgttat?gcttggttcc?actcggcgtg?ataagttatc
541?tatcgctgag?gacacttttt?ttaaaaaaag?gtggccaagg?taaatgcaga?tgaaccgctt
601?ccaatagtcc?atttgcttgg?acaataataa?tttttatgat?ctgatctcaa?atcaggtagg
661?actacccgct?gaacttaagc?atatcaataa?gcggagga
The rDNA-ITS base sequence of said rhizoctonia (Rhizoctonia sp.) GXSH-6 is following:
1 tccgtaggtg?aacctgcgga?aggatcatta?atgaatttaa?tgtggagttg?gttgtagctg
61 gtccgctgga?tttctttttt?taaaaaaaat?aaattcctct?gggcatgtgc?acgccttctc
121?tttcatccac?acacacctgt?gcacctgtga?ggcagatgtg?ggcacttgat?ctgtgcccct
181?ctgtctactt?aattcacaca?aactcaattt?atcttaaaat?gaatgtaatt?gatgtaacgc
241?atctaatact?aagtttcaac?aacggatctc?ttggctctcg?catcgatgaa?gaacgcagcg
301?aaatgcgata?agtaatgtga?attgcagaat?tcagtgaatc?atcgaatctt?tgaacgcacc
361?ttgcgctcct?tggtattcct?tggagcatgc?ctgtttgagt?atcatgaaat?cttcaaagtc
421?aatcttttgt?taattcaact?ggttgtgctt?tggtattgga?ggtctttgca?gcttcacacc
481?tgctcctctt?tgttcattag?ctggatctca?gtgttatgct?tggttccact?cggcgtgata
541?agttatctat?cgctgaggac?accgtaaaag?gtggccaagg?taaacacaga?ttgaaccgct
601?tctaatagtc?catttacttg?gacaacatat?ttatgatttg?atctcaaatc?aggtaggact
661?acccgctgaa?cttaagcata?tcaataagcg?gagga
Sequence table
< 110>Dongguan City Inst. of Biological Technology, Agricultural University Of South China
< 120>application of a kind of Herba Anoectochili roxburghii endogenetic fungus in Herba Anoectochili roxburghii is grown seedlings
<140>200910210073.2
On November 4th, < 141>2009
<160>3
<210>1
<211>693
<212>DNA
< 213>rhizoctonia (Rhizoctonia sp.) JSH-1
<400>1
tccgtaggtg?aacctgcgga?aggatcatta?ttgaatttta?atgaggagtt?ggttgttgct 60
ggccttggga?tttttttaac?taattattcc?aggcacttgt?gcacaccttc?tctttcatcc 120
acacacacct?gtgcacttgt?gagacagact?gtggggactt?tattgaaccc?cgtctgtcta 180
cttaatttac?acacactcta?tttaatttaa?actgaatgta?attgatgtaa?cgcatctaat 240
actaagtttc?aacaacggat?ctcttggctc?tcgcatcgat?gaagaacgca?gcgaaatgcg 300
ataagtaatg?tgaattgcag?aattcagtga?atcatcgaat?ctttgaacgc?accttgcgct 360
ccttggtatt?ccttggagca?tgcctgtttg?agtatcatga?aatcttcaaa?gtaaaccttt 420
tgttaattca?atgggttctt?tactttggta?ttggaggtct?ttgcagtctc?acaactgctc 480
ctctttgttc?attagctgga?tctcagtgtt?atgcttggtt?ccactcggcg?tgataagtat 540
ctatcgctga?ggacacctgt?aaaaaggtgg?ccaaggtaaa?tgcagatgaa?ccgcttctaa 600
cagtccattg?acttggacaa?ataaatattt?atgatctgat?ctcaaatcag?gtaggactac 660
ccgctgaact?taagcatatc?aataagcgga?gga 693
<210>2
<211>698
<212>DNA
< 213>rhizoctonia (Rhizoctonia sp.) GXSH-5
<400>2
tccgtaggtg?aacctgcgga?aggatcatta?atgaattaat?gtggagttgg?ttgtagctgg 60
tccgctggat?tttttaaaat?aaattccttt?ggacatgtgc?acgccttctc?tttcatccac 120
acacacctgt?gcacctgtga?ggcagttgtt?ggggacttta?ttgtaccctc?tctgtctact 180
taatttacac?aaactcaatt?tattttaaaa?cgaatgtaat?tgatgtaacg?catctaatac 240
taagtttcaa?caacggatct?cttggctctc?gcatcgatga?agaacgcagc?gaaatgcgat 300
aagtaatgtg?aattgcagaa?ttcagtgaat?catcgaatct?ttgaacgcac?cttgcgctcc 360
ttggtattcc?ttggagcatg?cctgtttgag?tatcatgaaa?tcttcaaagt?caatcttttt 420
gttaattcaa?ttggttgtgc?tttggtattg?gaggtctttt?gcagcttcac?acctgctcct 480
ctttgttcat?tagctggatc?tcagtgttat?gcttggttcc?actcggcgtg?ataagttatc 540
tatcgctgag?gacacttttt?ttaaaaaaag?gtggccaagg?taaatgcaga?tgaaccgctt 600
ccaatagtcc?atttgcttgg?acaataataa?tttttatgat?ctgatctcaa?atcaggtagg 660
actacccgct?gaacttaagc?atatcaataa?gcggagga 698
<210>3
<211>695
<212>DNA
< 213>rhizoctonia (Rhizoctonia sp.) GXSH-6
<400>3
tccgtaggtg?aacctgcgga?aggatcatta?atgaatttaa?tgtggagttg?gttgtagctg 60
gtccgctgga?tttctttttt?taaaaaaaat?aaattcctct?gggcatgtgc?acgccttctc 120
tttcatccac?acacacctgt?gcacctgtga?ggcagatgtg?ggcacttgat?ctgtgcccct 180
ctgtctactt?aattcacaca?aactcaattt?atcttaaaat?gaatgtaatt?gatgtaacgc 240
atctaatact?aagtttcaac?aacggatctc?ttggctctcg?catcgatgaa?gaacgcagcg 300
aaatgcgata?agtaatgtga?attgcagaat?tcagtgaatc?atcgaatctt?tgaacgcacc 360
ttgcgctcct?tggtattcct?tggagcatgc?ctgtttgagt?atcatgaaat?cttcaaagtc 420
aatcttttgt?taattcaact?ggttgtgctt?tggtattgga?ggtctttgca?gcttcacacc 480
tgctcctctt?tgttcattag?ctggatctca?gtgttatgct?tggttccact?cggcgtgata 540
agttatctat?cgctgaggac?accgtaaaag?gtggccaagg?taaacacaga?ttgaaccgct 600
tctaatagtc?catttacttg?gacaacatat?ttatgatttg?atctcaaatc?aggtaggact 660
acccgctgaa?cttaagcata?tcaataagcg?gagga
Claims (4)
1. the application of Herba Anoectochili roxburghii endogenetic fungus in Herba Anoectochili roxburghii is grown seedlings is characterized in that: this endogenetic fungus is rhizoctonia (Rhizoctonia sp.) JSH-1; This bacterial strain has been preserved in China Microbial Culture Preservation Commission common micro-organisms center, and its deposit number is CGMCC NO.3074, preservation date on May 20th, 2009.
2. Herba Anoectochili roxburghii endogenetic fungus according to claim 1 is cultivated the application in the seedling in the Herba Anoectochili roxburghii group, it is characterized in that: anoectochilus symbiosis seedling is cultivated in said rhizoctonia (Rhizoctonia sp.) JSH-1 and the symbiosis of Herba Anoectochili roxburghii tissue cultured seedling.
3. Herba Anoectochili roxburghii endogenetic fungus according to claim 2 is cultivated the application in the seedling in the Herba Anoectochili roxburghii group, it is characterized in that: the mode that said symbiosis is cultivated is for putting in order strain and said rhizoctonia (Rhizoctonia sp.) JSH-1 co-cultivation with Herba Anoectochili roxburghii group training seedling.
4. cultivate the application in the seedling according to the said Herba Anoectochili roxburghii endogenetic fungus of claim 3 in the Herba Anoectochili roxburghii group, it is characterized in that: said co-cultivation is in symbiotic culture medium, under the condition that is hedged off from the outer world, to carry out; Perhaps in cultivation medium, carry out under the open condition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102100732A CN101953261B (en) | 2009-11-04 | 2009-11-04 | Application of jewel orchid endophytic fungi to jewel orchid seedlings |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102100732A CN101953261B (en) | 2009-11-04 | 2009-11-04 | Application of jewel orchid endophytic fungi to jewel orchid seedlings |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101953261A CN101953261A (en) | 2011-01-26 |
| CN101953261B true CN101953261B (en) | 2012-05-30 |
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| CN103202227A (en) * | 2013-03-22 | 2013-07-17 | 赖新权 | Seedling breeding and rapid propagation cultivation method for traditional Chinese herbal medicine anoectochilus formosanus |
| CN104137727B (en) * | 2014-07-28 | 2016-03-30 | 厦门加晟生物科技有限公司 | A kind of implantation methods of roxburgh anoectochilus terminal bud |
| CN113025496B (en) * | 2019-12-24 | 2022-03-15 | 中国医学科学院药用植物研究所 | Fungus for inducing anoectochilus formosanus seeds to germinate into seedlings |
| CN116076344A (en) * | 2022-10-18 | 2023-05-09 | 广西壮族自治区中国科学院广西植物研究所 | Method for promoting differentiation, mycorrhizal breeding and cultivation of anoectochilus formosanus protocorm by utilizing rhizoctonia cerealis |
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| CN1433670A (en) * | 2002-01-25 | 2003-08-06 | 中国医学科学院药用植物研究所 | Application technology of mycorrhizal fungi in Anoectochilus blume cultivation |
| CN1961651A (en) * | 2005-11-07 | 2007-05-16 | 中国医学科学院药用植物研究所 | Application of mycorrhizal fungi in large-scale plantation of gold thread lotus |
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| CN1433670A (en) * | 2002-01-25 | 2003-08-06 | 中国医学科学院药用植物研究所 | Application technology of mycorrhizal fungi in Anoectochilus blume cultivation |
| CN1961651A (en) * | 2005-11-07 | 2007-05-16 | 中国医学科学院药用植物研究所 | Application of mycorrhizal fungi in large-scale plantation of gold thread lotus |
Non-Patent Citations (1)
| Title |
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| 唐明娟等.内生真菌对离体培养的福建金线莲生长的影响.《中草药》.2008,第39卷(第12期),1876-1880. * |
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