CN105838615A - Ustilago esculenta haploid strain UET2 and use thereof - Google Patents
Ustilago esculenta haploid strain UET2 and use thereof Download PDFInfo
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Abstract
The invention discloses an Ustilago esculenta haploid strain UET2 and a use thereof and belongs to the technical field of biology. The Ustilago esculenta haploid strain UET2 has a preservation number of CGMCC No. 11844. Zizania aquatica artificial breeding needs common invasion of two sexual compatible haploid strains. The strain can be used as a zizania aquatica artificial breeding mother strain, through screening, a sexual compatible haploid strain is obtained and artificial zizania aquatica breeding is realized through artificial inoculation. The haploid strain UEMT2 can be used for gene engineering reconstruction and provides an implementation base material for zizania aquatica improvement such as zizania aquatica environmental adaptability and zizania aquatica growth time control in zizania aquatica breeding.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of wild rice smut haploid strains UET2 and application thereof.
Background technology
Wild rice Ustilago, in Basidiomycotina Ustilago, is a kind of typical dichotype fungus, with the nearly source of Semen Maydis tumor smut.So far, Caulis Zizaniae caduciflorae is unique host that it is known, and the pregnant hay of Caulis Zizaniae caduciflorae is wild rice smut and the synergistic result of Caulis Zizaniae caduciflorae plant of this specificity parasitism.So far, the plantation of Caulis Zizaniae caduciflorae and conservation breeding still use the mode of hay pier separation screening, and manpower and materials put into relatively big, and wild rice smut exists multiple biological strain, and it may be mixed in Caulis Zizaniae caduciflorae plant, cause Zizania latifolia Cultivars Character instability, degenerate substantially.Therefore employing artificial vaccination mode is the developing direction of Caulis Zizaniae caduciflorae breeding.
Research shows that the two individual characteies affine monoploid combined inoculation of wild rice smut can make the wild pregnant hay of hay plant normally bloomed, i.e. plants stems base portion expands and no longer heading is bloomed, rather than the mixing of property affine haploid strains, nucleated mycelium or teleutospore inoculation all cannot make the wild pregnant hay of hay plant (the Semen Maydis tumor smut progress in nearly source also shows that the needs two affine monoploid of individual character of infecting of smut are mixed together inoculation), therefore, the affine haploid acquisition of wild rice smut property is the premise ensureing Caulis Zizaniae caduciflorae biological engineering.And wild rice smut exists with diploid teleutospore or nucleated mycelium in nature, there is no haploid strains.
Summary of the invention
The problem existed for prior art, it is an object of the invention to design provides a kind of wild rice smut haploid strains UET2 and the technical scheme of application thereof.
The present invention provide a kind of wild rice smut (Ustilago esculenta) haploid strains UET2 can promote the artificial pregnant hay of Caulis Zizaniae caduciflorae and Zizania latifolia Cultivars improvement.This bacterial strain is in December in 2015 07 day in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) preservation, and preserving number is CGMCC No.11844.Depositary institution address is: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.This bacterial strain has the property that
Morphological characteristic: rod-short, ripe spore is at 15-20 microns, and monokaryon, without barrier film, carries out budding (Fig. 2), like yeast, but bigger than yeast.
Cultural character: solid culture (YEPS solid medium: 2% peptone, 2% sucrose, 1% yeast powder, 1.5% agarose) bacterium colony is milk yellow, smooth surface is more moistening, thicker more greatly than bacterial clump, ratio antibacterial thickness during picking, similar yeast colony.Liquid culture (YEPS fluid medium: 2% peptone, 2% sucrose, 1% yeast powder) bacterium solution afterwards is homogeneous, such as antibacterial bacterium solution.
Physiological metabolism characteristic: the carbon nitrogen source that this bacterial strain preference reducing sugar (Fig. 4) and organic nitrogen source (Fig. 5) grow as it.
Cultural method: can In vitro culture, be suitable to carry out single bacterium colony on PDA or YEPS solid medium streak culture, cultivate in liquid culture and be easily reduced this bacterium activity.Additionally this bacterium is not suitable for subculture more than three times, and the cultivation continuously or the low temperature that are not suitable for more than 7 days are placed, and otherwise easily causes activity and reduces or thalline degraded, suitably at-80 degree 20% glycerol stocks.
Propagation method: budding.
Beneficial effects of the present invention: Caulis Zizaniae caduciflorae biological engineering needs jointly infecting of two kinds of affine haploid strains of property, this bacterial strain can be as the parent strain of Caulis Zizaniae caduciflorae biological engineering, and after screening obtains the haploid strains that its property is affine, i.e. can be realized artificial pregnant hay by Inoculation Method.This haploid strains can carry out genetic engineering modified thus improve for ensuing Zizania latifolia Cultivars simultaneously, provides practice processes material as improved the Caulis Zizaniae caduciflorae breeding work such as the environmental suitability of Caulis Zizaniae caduciflorae, control knot hay time.
Accompanying drawing explanation
Fig. 1 is UET2 haploid strains access approaches schematic diagram；
Fig. 2 is the haploid strains budding aspect graph of nuclear location EGFP protein overexpression；
Fig. 3 is haploid strains solid culture aspect graph；
Fig. 4 is haploid strains growth rate variation diagram in different carbon source culture medium；
Fig. 5 is haploid strains growth rate variation diagram in different nitrogen sources culture medium；
Fig. 6 is that during pregnant hay, Caulis Zizaniae caduciflorae plant stem apex tissue slice fluorescence microscopy observes figure after wild hay plant artificial vaccination wild rice smut, and containing UET2 bacterial strain during wherein left side figure is inoculation liquid, the right figure is without UET2 bacterial strain in inoculation liquid；
Fig. 7 is the PCR qualification result figure of strain and mating type gene.
Detailed description of the invention
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1: bacterial strain screening method
1. collection teleutospore: (Figure 1A) directly picking teleutospore from dragon No. 2 grey hays of hay.
2. teleutospore suspension: be placed in by the teleutosorus of picking in 5 mL sterilized water, filters through 4 layers of sterile gauze, it is thus achieved that purer teleutospore suspension, with sterilized water, teleutospore suspension is diluted to final concentration of 10 the most again3Individual spore/mL.Teleutospore form is as shown in Figure 1B.
3. teleutospore is cultivated: takes 100 L teleutospore suspensions and coats on basidiospore isolation medium, cultivates about 60 h, with scattered single bacterium colony (Fig. 1 C) tiny seen from naked eyes as standard for 28 DEG C.Basidiospore isolation medium (1 L) formula is: K2HPO4
1 g, MgSO4·7H2O 0.5 g, FeSO4·7H2O 0.01g, KCl 0.5 g, glucose 18 g, (NH4)2SO4
5.28 g, agar 10 g.
4. single spore separation is cultivated: in the single macroscopic bacterium colony of picking to clear water, be diluted to 103Individual spore/mL, take every time 1 L be placed under micrurgy instrument draw single basidiospore (Fig. 1 D), by isolated basidiospore at YEPS(2% peptone, 2% sucrose, 1% yeast powder, 1.5% agar powder with capillary pipette) on solid medium, cultivate 4 d for 28 DEG C.One single bacterium colony separates 5 basidiospore, takes more than 3 single bacterium colonies.Colonial morphology after basidiospore cultivation is as referring to figure 1e.
5. microexamination: the colonial morphology optical microscope of Formation of basidiospore is carried out microexamination, if colony edge smooth (Fig. 1 F), can be initially identified as haploid strains, if edge creates mycelia (Fig. 1 G), then give up.
6. the affine bacterial strain screening of property: the haploid strains of Preliminary Identification is numbered (Fig. 1 E), and to be diluted to concentration respectively with YEPS fluid medium be OD600It it is the bacterium solution of 2.0.Test bacterial strain affine to property by fusion reaction and carry out Preliminary Identification.Fusion reaction experiment such as Fig. 1 H, detailed process is: first No. 1 bacterium solution is carried out on YEPS solid medium point sample, each point 1 L, and total point sample number is the haploid strains number of isolated.Afterwards remaining each bacterium solution is respectively taken 1 L and is covered each by above No. 1 bacterial strain bacterium point, labeled strain number, in 28 DEG C of incubators, cultivate 4 d.After cultivation, colonial morphology being carried out perusal, if visible white aerial hyphae, then two bacterial strains being mixed may be the affine haploid strains of property.As shown in figure ih, No. 1 bacterial strain and 2,3,4,8 or No. 11 bacterial strains may be the affine haploid strains of property.
7. PCR identifies: by the clonal analysis to its Mating type locus, finds to exist in wild rice smut 3 trail receptorpraGene, is respectivelypra1、pra2Withpra3,pra1Nucleotide sequence as shown in SEQ ID NO:1,pra2Nucleotide sequence as shown in SEQ ID NO:2,pra3Nucleotide sequence as shown in SEQ ID NO:3.It is known that a haploid strains is only possible to have onepraGene, and in the affine haploid strains of two individual characteiespraGene is different.Therefore design specific primer carries out PCR checking for the monoploid obtained screening further and further confirms that sexual compatibility to each other.
As a example by above-mentioned 12 haploid strains screened, we extract the DNA of each bacterial strain, first pass through primer I TS1 and ITS4 amplification ITS sequence and order-checking carries out strain identification, and as illustrated in fig. 7d, sequencing result shows to belong to after NCBI comparison PCR resultUstilago esculenta。
Next in each strain gene group DNApra1、pra2Withpra3Carry out PCR amplification, result such as Fig. 7 A, shown in B, C, show that the trail receptor gene of 2, No. 4 bacterial strains ispra3, the trail receptor gene of 3,8, No. 11 bacterial strains ispra1, the trail receptor gene of remaining bacterial strain ispra2, the bacterial strain 1 trail receptor gene that i.e. our screening obtains ispra2, different with the genotype of 2,3,4,8 or No. 11 bacterial strains, therefore No. 1 bacterial strain and 2,3,4,8 or No. 11 bacterial strains are the affine haploid strains of property.Pcr amplification reaction system is: 10 × PCR buffer 5 μ L, dNTP 4 μ L, forward primer 1 μ L, downstream primer 1 μ L, DNA profiling 1 μ L, Taq enzyme 0.5 μ L, adds ddH2O to 50 μ L.PCR amplification program is 94 DEG C of 4 min；94 DEG C of 30 s, 55 DEG C of 30 s, 72 DEG C of 2 min, 30 circulations；72 ℃ 10 min；4 DEG C of terminations, expanding fragment length is all upper and lower at 600 bp, and primer sequence is as follows:
ITS1:5 '-TCCGTAGGTGAACCTGCGG-3 ' (nucleotide sequence is as shown in SEQ ID NO:4),
ITS4:5 '-TCCTCCGCTTATTGATATGC-3 ' (nucleotide sequence is as shown in SEQ ID NO:5),
Pra1-F:5 '-ATCGGCATCCTCGCTCATTATG-3 ' (nucleotide sequence is as shown in SEQ ID NO:6),
Pra1-R:5 '-TGCATGCTTGATCTCCGTTGCG-3 ' (nucleotide sequence is as shown in SEQ ID NO:7),
Pra2-F:5 '-ACAGCACGCTTCCCACCTTTTC-3 ' (nucleotide sequence is as shown in SEQ ID NO:8),
Pra2-R:5 '-GACAAAGCAGCAGTGAACTGCC-3 ' (nucleotide sequence is as shown in SEQ ID NO:9),
Pra3-F:5 '-CACAATTCCCATCACGGTGCTC-3 ' (nucleotide sequence is as shown in SEQ ID NO:10),
Pra3-R:5 '-GAGCGAGAGCACTGATGGAAAG-3 ' (nucleotide sequence is as shown in SEQ ID NO:11).
8. determine haploid strains: after using primer pra2-F and pra2-R amplification, obtain length be defined as haploid strains UET2 at the bacterial strain of the 600 upper and lower amplified fragments of bp.
This bacterial strain has the property that
Morphological characteristic: rod-short, ripe spore is at 15-20 microns, and monokaryon, without barrier film, carries out budding (Fig. 2), like yeast, but bigger than yeast.
Cultural character: solid culture (YEPS solid medium: 2% peptone, 2% sucrose, 1% yeast powder, 1.5% agarose) bacterium colony is milk yellow, smooth surface is more moistening, thicker more greatly than bacterial clump, ratio antibacterial thickness during picking, similar yeast colony (Fig. 3).Liquid culture (YEPS fluid medium: 2% peptone, 2% sucrose, 1% yeast powder) bacterium solution afterwards is homogeneous, such as antibacterial bacterium solution.
Physiological metabolism characteristic: the carbon nitrogen source that this bacterial strain preference reducing sugar (Fig. 4) and organic nitrogen source (Fig. 5) grow as it.
Cultural method: can In vitro culture, be suitable to carry out single bacterium colony on PDA or YEPS solid medium streak culture, cultivate in liquid culture and be easily reduced this bacterium activity.Additionally this bacterium is not suitable for subculture more than three times, and the cultivation continuously or the low temperature that are not suitable for more than 7 days are placed, and otherwise easily causes activity and reduces or thalline degraded, suitably at-80 degree 20% glycerol stocks.
Propagation method: budding.
Embodiment 2: artificial pregnant hay is tested
The protoplast transformation technology mediated by PEG by the vector wild rice smut of process LAN EGFP green fluorescent protein (Ustilago esculenta) haploid strains UET2 and the affine bacterial strain of property thereof (choose No. 2 in embodiment 1, No. 3), make its process LAN EGFP green fluorescent protein, the process LAN bacterial strain of acquisition can be detected by fluorescence microscope, the bacterial strain of success process LAN EGFP green fluorescent protein can see green florescent signal, as shown in Figure 2 under fluorescence microscope.Then carrying out artificial vaccination experiment, concrete artificial vaccination process is as follows:
1) by two wild rice smut affine haploid strains of property in YEPS fluid medium 28 DEG C shake bacterium to OD600It is 1.0, centrifugal collection thalline, it is diluted to final concentration of OD with 0.5 × YEPS fluid medium600It is the bacterium solution of 3.0, is mixed for connecing bacterium by two bacterium solution；
2) the wild hay tubulose root with more than 3 complete internodes is carried out nursery, 25 DEG C of hot-house cultures 20 days, with sprout have more than 3 little seedling stage plant tubulose root as object of inoculation, and carry out Seedling base portion pricking hole process；
3) mixed bacteria liquid syringe syringe-like root step 1) obtained, until there being bacterium solution to overflow；
4) the inoculation Seedling after processing is pruned, and prevents too much transpiration from causing here plant withers, and then leaching is placed in remaining mixed bacteria liquid, and 12 h are placed in 25 DEG C of greenhouses；
5) the wild hay Seedling after leaching bacterium is transferred in the little basin of band Nutrition Soil carry out transition inoculation, after soil is sufficiently humidified so as to, mixed bacteria liquid is poured in soil, 25 DEG C of hot-house culture 7 d, complete inoculation；
6) will carry out outdoor cultivation in postvaccinal Seedling transplanting outdoor or incubator, inoculation planting time controls in March, applies fertilizer and plants standard with reference to normal Caulis Zizaniae caduciflorae.
Result we have found that this bacterial strain infects wild Caulis Zizaniae caduciflorae plant together with its affine bacterial strain of property 3 and wild Caulis Zizaniae caduciflorae plants stems base portion can be made to expand formation Caulis Zizaniae caduciflorae, and No. 2 and No. 3 property affine bacterial strains mixing (i.e. after deletion mycopremna UET2) wild Caulis Zizaniae caduciflorae plants stems base portions no longer expand, i.e. cannot pregnant hay.Caulis Zizaniae caduciflorae plant shoot tip meristem sampling to inoculation latter two phenotype, find at fluorescence microscopy Microscopic observation after carrying out tissue slice: the plant that basal part of stem expands can substantially observe the mycelia with green fluorescence, and plant stem apex redgreen fluorescence signal (Fig. 6) do not expanded, show that this pregnant hay process needs the participation of UET2, i.e. haploid strains UET2 can be applicable to artificial pregnant hay.
SEQUENCE LISTING
<110>China Measures Institute
<120>a kind of wild rice smut (Ustilago esculenta) haploid strains UET2 and application thereof
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PatentIn version 3.3
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<212> DNA
<213>synthetic
<400> 6
atcggcatcc tcgctcatta tg
22
<210> 7
<211> 22
<212> DNA
<213>synthetic
<400> 7
tgcatgcttg atctccgttg cg
22
<210> 8
<211> 22
<212> DNA
<213>synthetic
<400> 8
acagcacgct tcccaccttt tc
22
<210> 9
<211> 22
<212> DNA
<213>synthetic
<400> 9
gacaaagcag cagtgaactg cc
22
<210> 10
<211> 22
<212> DNA
<213>synthetic
<400> 10
cacaattccc atcacggtgc tc
22
<210> 11
<211> 22
<212> DNA
<213>synthetic
<400> 11
gagcgagagc actgatggaa ag
22
Claims (3)
1. a wild rice smut (Ustilago
esculenta) haploid strains UET2, preserving number is: CGMCC No.11844.
2. wild rice smut as claimed in claim 1 a kind of (Ustilago
esculenta) haploid strains UET2 application in the artificial pregnant hay of Caulis Zizaniae caduciflorae.
3. wild rice smut as claimed in claim 1 a kind of (Ustilago esculenta) haploid strains UET2 application in Zizania latifolia Cultivars is improved.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108998464A (en) * | 2018-07-25 | 2018-12-14 | 中国计量大学 | PSP107 plasmid and its application, construction method |
| CN109370922A (en) * | 2018-12-18 | 2019-02-22 | 中国计量大学 | A pair of wild rice smut for successfully realizing the normal hay artificially breeding of wild rice stem and its application |
| CN109392702A (en) * | 2018-12-18 | 2019-03-01 | 中国计量大学 | A kind of method of the normal wild rice stem of artificially breeding |
| CN109593769A (en) * | 2018-12-18 | 2019-04-09 | 中国计量大学 | Wild rice brand spores form related gene Itd1 and its application |
| CN109593772A (en) * | 2018-12-18 | 2019-04-09 | 中国计量大学 | CRISPR/Cas9 plasmid and its construction method, application method |
| CN110331223A (en) * | 2019-07-08 | 2019-10-15 | 浙江大学 | It is a kind of for identifying molecular labeling, primer pair, kit and the method for different wild rice stem types |
| CN114175979A (en) * | 2021-11-19 | 2022-03-15 | 扬州大学 | Method for producing normal water bamboo from male water bamboo by artificially inoculating smut bacteria of water bamboo |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560742A (en) * | 2015-01-14 | 2015-04-29 | 浙江省林业科学研究院 | Agrobacterium-mediated ustilago esculenta transformant strain as well as preparation method and application thereof |
-
2016
- 2016-01-28 CN CN201610058043.4A patent/CN105838615B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560742A (en) * | 2015-01-14 | 2015-04-29 | 浙江省林业科学研究院 | Agrobacterium-mediated ustilago esculenta transformant strain as well as preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| MEIKE PIEPENBRING ET AL.: "the generic position of ustilago maydis,ustilago scitaminea, and ustilago esculenta (ustilaginales)", 《MYCOLOGICAL PROGRESS》 * |
| 柯卫东等: "菰黑粉菌不同菌株比较研究", 《长江蔬菜》 * |
| 胡鹏等: "菰黑粉菌转录调节因子rbf1的克隆及表达分析", 《植保研究》 * |
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| CN108998464A (en) * | 2018-07-25 | 2018-12-14 | 中国计量大学 | PSP107 plasmid and its application, construction method |
| CN109370922A (en) * | 2018-12-18 | 2019-02-22 | 中国计量大学 | A pair of wild rice smut for successfully realizing the normal hay artificially breeding of wild rice stem and its application |
| CN109392702A (en) * | 2018-12-18 | 2019-03-01 | 中国计量大学 | A kind of method of the normal wild rice stem of artificially breeding |
| CN109593769A (en) * | 2018-12-18 | 2019-04-09 | 中国计量大学 | Wild rice brand spores form related gene Itd1 and its application |
| CN109593772A (en) * | 2018-12-18 | 2019-04-09 | 中国计量大学 | CRISPR/Cas9 plasmid and its construction method, application method |
| CN109370922B (en) * | 2018-12-18 | 2020-10-16 | 中国计量大学 | Ustilago esculenta for successfully realizing artificial breeding of normal zizania aquatica and application thereof |
| CN109392702B (en) * | 2018-12-18 | 2022-09-20 | 中国计量大学 | Method for artificially breeding normal cane shoots |
| CN110331223A (en) * | 2019-07-08 | 2019-10-15 | 浙江大学 | It is a kind of for identifying molecular labeling, primer pair, kit and the method for different wild rice stem types |
| CN110331223B (en) * | 2019-07-08 | 2021-06-01 | 浙江大学 | Molecular marker, primer pair, kit and method for identifying different cane shoots types |
| CN114175979A (en) * | 2021-11-19 | 2022-03-15 | 扬州大学 | Method for producing normal water bamboo from male water bamboo by artificially inoculating smut bacteria of water bamboo |
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