CN108588269A - A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness - Google Patents
A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness Download PDFInfo
- Publication number
- CN108588269A CN108588269A CN201810751513.4A CN201810751513A CN108588269A CN 108588269 A CN108588269 A CN 108588269A CN 201810751513 A CN201810751513 A CN 201810751513A CN 108588269 A CN108588269 A CN 108588269A
- Authority
- CN
- China
- Prior art keywords
- issr
- dendrobium candidum
- primer
- pcr
- dendrobidium huoshanness
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Botany (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of ISSR primers and its reaction system differentiating dendrobium candidum and Dendrobidium huoshanness, the primer nucleotide sequences carry out amplification to dendrobium candidum and Dendrobidium huoshanness DNA as shown in NO.1~6 SEQ ID, using the reaction system after the primer and optimization and can be completed.Primer of the present invention and method are simple, save time and cost, and have preferable application value to differentiating dendrobium candidum and Dendrobidium huoshanness, analyzing its genetic affinity etc..
Description
Technical field
The invention belongs to molecular marking technique fields, and in particular to a kind of to differentiate that the ISSR of dendrobium candidum and Dendrobidium huoshanness draws
Object and its reaction system.
Background technology
For Shihu " medicinal materials as rare Chinese medicine, medicinal history is long, derives from a wealth of sources, and it is extremely prominent to mix pseudo- phenomenon.Wherein suddenly
The mountain stem of noble dendrobium and dendrobium candidum often mutually mix puppet because of the similitude of morphosis.Dendrobium candidum (Dendrobium officinale
Kimura et Migo) it is orchid family Dendrobium rare kind the most rare, medicinal part is fresh or dry stem, and there is beneficial stomach to give birth to
Tianjin nourishing Yin and clearing heat, is moistened the lung and relieve the cough, improving eyesight salubrity and other effects.Dendrobidium huoshanness (Dendrobium huoshanense
C.Z.TangetS.J.Cheng), also known as the suddenly stem of noble dendrobium, suddenly dry measure used in former times, rice dry measure used in former times, the Dabie Mountain stem of noble dendrobium, be orchid species at most and distribution
One of most wide category.The growth cycle of dendrobium candidum is slow, and growth conditions is harsh, and the market demand is big, thus adulterant occurs.And
Dendrobidium huoshanness is rare rare, leads to the phenomenon that pretending to be with dendrobium candidum often occur.Therefore, the identification system of efficient quick is built
It is of great significance for the identification and protection of resources of dendrobium candidum and Dendrobidium huoshanness.
Molecular marking technique can reflect the hereditary difference of species from DNA level, be to study genetic diversity to have efficacious prescriptions
Method.ISSR is a kind of Inter-simple sequence repeat molecular labeling, because its is simple, convenient and rapid, polymorphism is high, repeated
The advantages that good, is widely used in the cultivar identification etc. of animals and plants.But ISSR labels are a kind of randomness labeling methods, not jljl
Its test parameters is variant under kind even same species different condition, therefore establishes a scientific and reasonable ISSR-PCR reaction
System seeks the dosage of best PCR each components, can be that ISSR labels is further utilized to differentiate dendrobium candidum and Dendrobidium huoshanness
Theoretical foundation is provided.
Invention content
The object of the present invention is to provide the ISSR-PCR molecule marks of discriminating dendrobium candidum and Dendrobidium huoshanness after a kind of optimization
Note system.
The present invention is realized especially by following technical scheme：
It is a kind of differentiate dendrobium candidum and Dendrobidium huoshanness ISSR primers, the ISSR primers include UBC817, UBC826,
Six specific primers of UBC827, UBC834, UBC842 or UBC849, specially：
UBC817：CACACACACACACACAA；
UBC826：ACACACACACACACACC；
UBC827：ACACACACACACACACG；
UBC834：AGAGAGAGAGAGAGAGYT；
UBC842：GAGAGAGAGAGAGAGAYG；
UBC849：GTGTGTGTGTGTGTGTYA.
The present invention also provides a kind of reaction systems differentiating dendrobium candidum and Dendrobidium huoshanness, specifically include following steps：
1) tender leaf for acquiring dendrobium candidum and Dendrobidium huoshanness, for extracting plant group DNA；
2) integrality of 1% agarose gel electrophoresis detection DNA is used；
3) DNA sample for using ISSR-PCR reaction systems to extract step (1) expands.
Described ISSR-PCR reactions primer used be UBC817, UBC826, UBC827, UBC834, UBC842 or
UBC849。
The amplification system of the ISSR-PCR is：Contain 2 × TaqMaster Mix, 11.5 μ in the reaction system of 20 μ L
L, template DNA 35ng, 0.4 μM of primer.
The amplification program of the ISSR-PCR is：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 52.2~56.2 DEG C are moved back
Fiery 30s, 72 DEG C of extension 1.5min are recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
The present invention also provides above-mentioned primer UBC817, UBC826, UBC827, UBC834, UBC842 and UBC849 to reflect
Application in other dendrobium candidum and Dendrobidium huoshanness.
Beneficial effects of the present invention are：
The present invention provides a kind of ISSR primers of discriminating dendrobium candidum and Dendrobidium huoshanness, and is established on the basis of the primer
The system for differentiating dendrobium candidum and Dendrobidium huoshanness using ISSR molecular labelings, according to the results show that system amplification band
It is steady and audible, have the characteristics that preferable polymorphism, operating procedure is less, is easy and fast to analysis detection.
Description of the drawings
Fig. 1 is the amplification of ISSR-PCR Uniform Designs of the present invention；Wherein M is indicated：marker；DNA profiling is
Dendrobium candidum；Primer is UBC834；1-12 is that 1-12 is combined in the processing of table 3；
Fig. 2 is ISSR-PCR amplifications of the present invention 2 × Taq Master Mix as single factor test；M is indicated：
marker；DNA profiling is dendrobium candidum；Primer is UBC834；The amount of 2 × TaqMaster Mix is from left to right followed successively by：12μ
L, 10 μ L, 8 μ L, 6 μ L, 4 μ L, 2 μ L；
Fig. 3 is the ISSR-PCR amplifications of the present invention 2 × Taq Master Mix fine tunings；M is indicated：marker；DNA moulds
Plate is dendrobium candidum；Primer is UBC834；The amount of 2 × Taq MasterMix is from left to right followed successively by：10 μ L, 10.2 μ L, 10.4
μ L, 10.6 μ L, 10.8 μ L, 11 μ L, 11.2 μ L, 11.4 μ L, 11.6 μ L, 11.8 μ L；
Fig. 4 is ISSR-PCR amplification of the primer of the present invention as single factor test；M is indicated：marker；DNA profiling is iron
The skin stem of noble dendrobium；Primer is UBC834；The amount of primer is from left to right followed successively by：1 μ L, 2 μ L, 3 μ L, 4 μ L, 5 μ L；
Fig. 5 is the ISSR-PCR amplifications of primer of the present invention fine tuning；M is indicated：marker；DNA profiling is dendrobium candidum；
Primer is UBC834；The amount of primer is from left to right followed successively by：2.8 μ L, 3 μ L, 3.2 μ L, 3.4 μ L, 3.6 μ L, 3.8 μ L, 4 μ L, 4.2
μL；
Fig. 6 is ISSR-PCR amplifications of the DNA of the present invention as single factor test；M is indicated：marker；DNA profiling is iron sheet
The stem of noble dendrobium；Primer is UBC834；The amount of DNA is from left to right followed successively by：0.5 μ L, 1 μ L, 1.5 μ L, 2 μ L, 2.5 μ L, 3 μ L, 3.5 μ L；
Fig. 7 is the ISSR-PCR amplifications of discriminating dendrobium candidum and Dendrobidium huoshanness after present invention optimization；M is indicated：
marker；DNA profiling is dendrobium candidum and Dendrobidium huoshanness；Primer is from left to right followed successively by：UBC826, UBC827, UBC834,
UBC842, UBC849, UBC817.
Specific implementation mode
Below in conjunction with specific embodiment of the present invention, technical solution of the present invention is clearly and completely described, is shown
So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
The foundation and optimization of embodiment 1ISSR-PCR amplification systems
1, the extraction of DNA
By mortar clear water clean dry, lighted after the absolute ethyl alcohol of 5ml is added, after mortar dry in the air it is cool after, take collected
Dendrobium candidum and Dendrobidium huoshanness tender shoots 500mg pour into appropriate liquid nitrogen in mortar, do not have sample 2~3cm height to be with liquid nitrogen
Preferably, 3~5min is ground, then RNA isolation kit is used to extract stem of noble dendrobium genomic DNA.
2, the foundation of ISSR-PCR amplification systems
1) the Uniform Design scheme of ISSR-PCR reaction systems
Using U12(43) uniform designs table, the factor level of PCR amplification system is shown in Table 2.Using UBC834 as primer, iron sheet stone
The genomic DNA of dry measure used in former times is template, optimizes screening to ISSR-PCR using 12 reaction systems in table 3, reaction system is
20 μ L, are specifically shown in Table 3.
ISSR-PCR amplification programs are：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 53.6 DEG C of annealing 30s, 72 DEG C extend
1.5min is recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
The primer and sequence that table 1 screens
2 factor level table (unit of table：μl)
3 Uniform Design table U of table12(43)/unit：μ
The results are shown in Figure 1, and in addition to 7,9, other processing have amplified band.Wherein processing 2,3 has three amplified bands,
Other processing only have two amplified bands, and the band of processing 3 is clear compared with the band of processing 2, this shows processing 3 more to manage
The amplification condition thought.
2) ISSR-PCR testing programs of 2 × Taq Master Mix as single factor test
According to Uniform Design as a result, using UBC834 as primer, the genomic DNA of dendrobium candidum is template, in 20 μ L
Reaction system in, the amount of design 2 × Taq Master Mix is respectively that 2 μ l, 4 μ l, 6 μ l, 8 μ l, 10 μ l, 12 μ l are carried out
ISSR-PCR, primer, DNA and ddH2The dosage of O is shown in Table 4.
ISSR-PCR amplification programs are：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 53.6 DEG C of annealing 30s, 72 DEG C extend
1.5min is recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
Single factor experiment table (the unit of table 42 × Taq Master Mix：μl)
The single factor experiment result of 2 × Taq Master Mix is shown in Fig. 2, and processing 4,5,6 has amplified band, and processing 5 has
Three amplified bands, this shows that processing 5 is ideal amplification condition.
3) 2 × Taq Master Mix finely tune testing program as the ISSR-PCR of single factor test
According to Uniform Design as a result, using UBC834 as primer, the genomic DNA of dendrobium candidum is template, in 20 μ L
Reaction system in, the amount of design 2 × Taq Master Mix be respectively 10 μ l, 10.2 μ l, 10.4 μ l, 10.6 μ l, 10.8 μ l,
11 μ l, 11.2 μ l, 11.4 μ l, 11.6 μ l, 11.8 μ l carry out ISSR-PCR, primer, DNA and ddH2The dosage of O is shown in Table 5.
ISSR-PCR amplification programs are：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 53.6 DEG C of annealing 30s, 72 DEG C extend
1.5min is recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
Table 52 × Taq Master Mix fine tuning experiment table (units：μl)
2 × Taq Master Mix fine tunings test result is shown in that Fig. 3, all processing have amplified band.Processing 8,9,10
Amplified band is apparent, is ideal amplification condition.
4) ISSR-PCR testing program of the primer as single factor test
Using UBC834 as primer, the genomic DNA of dendrobium candidum is template, in the reaction system of 20 μ L, design primer
Amount be respectively that 1 μ l, 2 μ l, 3 μ l, 4 μ l, 5 μ l carry out ISSR-PCR, 2 × Taq Master Mix, DNA and ddH2The dosage of O
It is shown in Table 6.
ISSR-PCR amplification programs are：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 53.6 DEG C of annealing 30s, 72 DEG C extend
1.5min is recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
Single factor experiment table (the unit of 6 primer of table：μl)
Primer single factor experiment result is shown in that Fig. 4, all processing have amplified band.Handle 1,2 band numbers relatively processing 3,4,
5, illustrate that processing 3,4,5 is ideal amplification condition.
5) primer finely tunes testing program as the ISSR-PCR of single factor test
Using UBC834 as primer, the genomic DNA of dendrobium candidum is template, in the reaction system of 20 μ L, design primer
Amount be respectively that 2.8 μ l, 3 μ l, 3.2 μ l, 3.4 μ l, 3.6 μ l, 3.8 μ l, 4 μ l carry out ISSR-PCR, 2 × Taq Master
Mix, DNA and ddH2The dosage of O is shown in Table 7.
ISSR-PCR amplification programs are：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 53.6 DEG C of annealing 30s, 72 DEG C extend
1.5min is recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
7 primer of table fine tuning experiment table (unit：μl)
Primer fine tuning test result is shown in Fig. 5, and all processing have band, and 3,4,5,6,7 bands of processing are more clear, are
Comparatively ideal amplification condition.
6) ISSR-PCR testing programs of the DNA as single factor test
Using UBC834 as primer, the genomic DNA of dendrobium candidum is template, in the reaction system of 20 μ L, design dna
Amount is respectively 0.5 μ l, 1 μ l, 1.5 μ l, 2 μ l, 2.5 μ l, 3 μ l, 3.5 μ l progress ISSR-PCR, 2 × Taq Master Mix, DNA
And ddH2The dosage of O is shown in Table 8.
ISSR-PCR amplification programs are：94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation 30s, 53.6 DEG C of annealing 30s, 72 DEG C extend
1.5min is recycled 35 times, and 72 DEG C re-extend 10min, 4 DEG C of preservations.
Single factor experiment table (the unit of table 8DNA：μl)
The single factor experiment result of DNA is shown in Fig. 6, and all processing have band, and it is apparent to handle 3,6,7 bands, for compared with
Ideal amplification condition.
Embodiment 2
Dendrobium candidum source：The hall agricultural development Co., Ltd of Daye health.
Dendrobidium huoshanness source：Chinese medicine Dendrobidium huoshanness Science and Technology Ltd..
Using the reaction system after optimization, with UBC826, UBC827, UBC834, UBC842, UBC849, UBC817 is to draw
The DNA sample of object, dendrobium candidum and Dendrobidium huoshanness is template, optimizes the PCR verifications of system.Amplification program is：94 DEG C pre-
It is denaturalized 5min, 94 DEG C of denaturation 30s, 52.2~56.2 DEG C of annealing 30s, 72 DEG C of extension 1.5min, cycle 35 times, 72 DEG C re-extend
10min, 4 DEG C of preservations.
Annealing temperature：
Using the reaction system after optimization and primer UBC826, UBC827, UBC834, UBC842, UBC849 after screening,
UBC817 optimizes the PCR verifications of system using the DNA sample of dendrobium candidum and Dendrobidium huoshanness as template, the results showed that, expand
It is steady and audible to increase band, characteristic strip is apparent (Fig. 7, white box are characterized band), illustrates that the reaction system and selected primer are suitble to
The ISSR of dendrobium candidum and Dendrobidium huoshanness is analyzed.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding without departing from the principles and spirit of the present invention can carry out these embodiments a variety of variations, modification, replace
And modification, the scope of the present invention is defined by the appended.
Sequence table
<110>Research Institute of Traditional Chinese Medicine
<120>A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness
<141> 2018-07-10
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
cacacacaca cacacaa 17
<210> 2
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
acacacacac acacacc 17
<210> 3
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
acacacacac acacacg 17
<210> 4
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
agagagagag agagagyt 18
<210> 5
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
gagagagaga gagagayg 18
<210> 6
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
gtgtgtgtgt gtgtgtya 18
Claims (6)
1. a kind of ISSR primers differentiating dendrobium candidum and Dendrobidium huoshanness, which is characterized in that the ISSR primers include
UBC817, UBC826, UBC827, UBC834, UBC842 or UBC849, nucleotide sequence is respectively such as SEQ ID NO.1~6 institutes
Show.
2. a kind of ISSR-PCR reaction systems differentiating dendrobium candidum and Dendrobidium huoshanness, which is characterized in that include the following steps：
1) tender leaf for acquiring dendrobium candidum and Dendrobidium huoshanness, for extracting plant group DNA；
2) integrality of 1% agarose gel electrophoresis detection DNA is used；
3) DNA sample for using ISSR-PCR reaction systems to extract step (1) expands.
3. reaction system according to claim 2, which is characterized in that described ISSR-PCR reactions primer used is
UBC817, UBC826, UBC827, UBC834, UBC842 or UBC849.
4. reaction system according to claim 2, which is characterized in that the amplification system of the ISSR-PCR is：20μL
Reaction system in contain 2 × Taq Master Mix 11.5 μ L, template DNA 35ng, 0.4 μM of primer.
5. reaction system according to claim 2, which is characterized in that the amplification program of the ISSR-PCR is：94℃
Pre-degeneration 5min, 94 DEG C of denaturation 30s, 52.2~56.2 DEG C of annealing 30s, 72 DEG C of extension 1.5min, cycle 35 times, 72 DEG C re-extend
10min, 4 DEG C of preservations.
6. application of the primer described in claim 1 in differentiating dendrobium candidum and Dendrobidium huoshanness.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810751513.4A CN108588269A (en) | 2018-07-10 | 2018-07-10 | A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810751513.4A CN108588269A (en) | 2018-07-10 | 2018-07-10 | A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108588269A true CN108588269A (en) | 2018-09-28 |
Family
ID=63618024
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810751513.4A Pending CN108588269A (en) | 2018-07-10 | 2018-07-10 | A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108588269A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110878376A (en) * | 2019-12-31 | 2020-03-13 | 安徽师范大学 | SSR molecular marker primer for identifying dendrobium huoshanense and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101451163A (en) * | 2008-12-22 | 2009-06-10 | 陈乃富 | Construction and identification method of molecular marking fingerprint of Dendrobium huoshanense and similitude species thereof |
| CN106498064A (en) * | 2016-11-08 | 2017-03-15 | 皖西学院 | Differentiate the ISSR fingerprint spectrum methods of three kinds of medicinal dendrobiums and its sibling species |
-
2018
- 2018-07-10 CN CN201810751513.4A patent/CN108588269A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101451163A (en) * | 2008-12-22 | 2009-06-10 | 陈乃富 | Construction and identification method of molecular marking fingerprint of Dendrobium huoshanense and similitude species thereof |
| CN106498064A (en) * | 2016-11-08 | 2017-03-15 | 皖西学院 | Differentiate the ISSR fingerprint spectrum methods of three kinds of medicinal dendrobiums and its sibling species |
Non-Patent Citations (6)
| Title |
|---|
| KULDEEP SHARMA等: "ISSR marker-assisted selection of male and female plants in a promising dioecious crop: jojoba (Simmondsia chinensis)", 《PLANT BIOTECHNOLOGY REPORTS》 * |
| 刘明珍等: "道地药材霍山石斛及其相似种的分子鉴定", 《生物学杂志》 * |
| 张梓浩: "铁皮石斛工厂化育苗技术研究及遗传稳定性分析", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 徐程: "铁皮石斛种质资源与组培工厂化生产研究", 《中国博士学位论文全文数据库 农业科技辑》 * |
| 李盛清等: "石斛ISSR-PCR体系的优化", 《江苏农业科学》 * |
| 沈颖等: "ISSR-PCR在石斛种间鉴别中的应用", 《中草药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110878376A (en) * | 2019-12-31 | 2020-03-13 | 安徽师范大学 | SSR molecular marker primer for identifying dendrobium huoshanense and application thereof |
| CN110878376B (en) * | 2019-12-31 | 2022-11-22 | 安徽师范大学 | SSR molecular marker primer for identifying dendrobium huoshanense and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106636342B (en) | Acquisition method and application of EST-SSR labeled primer group developed based on ligusticum wallichii transcriptome sequence | |
| CN107058508B (en) | Salvia miltiorrhiza germplasm resource identification method | |
| CN109055595A (en) | A kind of Herba Epimedii ISSR-PCR molecule labelling method | |
| CN111500762A (en) | Sagittaria trifolia SSR primer group and application thereof | |
| CN112695124B (en) | Phalaenopsis SSR molecular marker primer composition and application thereof | |
| CN113151567B (en) | SSR molecular marker and method for identifying Lepista sordida N006# strain | |
| CN112662806B (en) | Rhynchosia SSR molecular marker primer composition and application thereof | |
| CN106434988A (en) | Wu-he dipsacus asper ISSR-PCR molecule marking method | |
| CN108588269A (en) | A kind of ISSR primers and its method differentiating dendrobium candidum and Dendrobidium huoshanness | |
| CN107523637A (en) | The method for differentiating sika deer, red deer or its hybridization deer is established based on COI and SRY sequences | |
| CN110878376A (en) | SSR molecular marker primer for identifying dendrobium huoshanense and application thereof | |
| CN110205397B (en) | Wushan epimedium and EST-SSR molecular identification method of easily-mixed species thereof | |
| CN114807413A (en) | Olive ISSR-PCR molecular marker combination and application thereof | |
| CN106676176A (en) | Method for performing SSR analysis on tetraploid alfalfa by utilizing multiple PCR | |
| CN109762918B (en) | Characteristic sequence, primer and method for identifying polygonatum longstem and polygonatum cyrtonema | |
| CN112941223A (en) | Golden needle mushroom 6168 strain and identification method and construction method and application of SSR marker fingerprint spectrum thereof | |
| CN112980991B (en) | Identification method of SSR (simple sequence repeat) marker fingerprint of Shanghai Zhen 30 strain of hypsizigus marmoreus, construction method and application thereof | |
| CN112795682B (en) | SSR (simple sequence repeat) marker fingerprint of Shanghai Zhen 6 strain of hypsizigus marmoreus, construction method and application thereof | |
| CN112695131B (en) | SSR (simple sequence repeat) marker fingerprint of Shanghai Zhen 29 strain of hypsizigus marmoreus, construction method and application thereof | |
| CN112795685B (en) | SSR (simple sequence repeat) marker fingerprint of Shanghai Zhen 11 strain of hypsizigus marmoreus, construction method and application thereof | |
| CN112795688B (en) | SSR (simple sequence repeat) marker fingerprint spectrum of hypsizigus marmoreus HM13 strain as well as construction method and application thereof | |
| CN112813183B (en) | SSR marker fingerprint of Hypsizigus marmoreus Finc-B-6 strain and construction method and application thereof | |
| CN112760406B (en) | SSR (simple sequence repeat) marker fingerprint spectrum of hypsizigus marmoreus HM22 strain as well as construction method and application thereof | |
| CN112760407B (en) | SSR (simple sequence repeat) marker fingerprint of hypsizigus marmoreus HM37 strain, and construction method and application thereof | |
| CN112795687B (en) | SSR (simple sequence repeat) marker fingerprint spectrum of hypsizigus marmoreus HM21 strain as well as construction method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180928 |
|
| RJ01 | Rejection of invention patent application after publication |