CN116240310A - SSR molecular marker combination for evaluating genetic diversity of Choerospondias axillaris and application thereof - Google Patents
SSR molecular marker combination for evaluating genetic diversity of Choerospondias axillaris and application thereof Download PDFInfo
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Abstract
The invention discloses an SSR molecular marker combination for evaluating genetic diversity of Choerospondias axillaris and application thereof, and relates to the technical field of molecular genetics. The SSR molecular marker combination comprises molecular markers NSZ-43385, NSZ-9141, NSZ-37910, NSZ-240, NSZ-22900, NSZ-39803, NSZ-42088, NSZ-165, NSZ-26522, NSZ-39039, NSZ-41051, NSZ-1007, NSZ-36575, NSZ-37045, NSZ-34772 and NSZ-45650. The SSR molecular marker combination can be used for evaluating the genetic diversity of the Choerospondias axillaris, researching the genetic diversity of different groups of the Choerospondias axillaris, and has important guiding significance in the aspects of formulating effective protection and sustainable utilization strategies of choerospondias axillaris germplasm resources and the like.
Description
Technical Field
The invention relates to the technical field of molecular genetics, in particular to an SSR molecular marker combination for evaluating genetic diversity of Choerospondias axillaris and application thereof.
Background
The Choerospondias axillaris (Choerospondias axillaris (Roxb.) Burtt et Hill.) is a single species of Choerospondias axillaris belonging to Anaardiaceae (Anaardiaceae), has high nutritive value and rich components, and is a tree species with good economic, ecological and social benefits. The Choerospondias axillaris leaf can be used as fertilizer, the wood is wide in application, and the bark and She Kedi tannin extract are used as raw materials. The fruit can be eaten raw or used for brewing wine. The kernel can be used as active carbon raw material. The bark fiber can be used as rope. The bark and fruit are used as medicines, and have the effects of diminishing inflammation, detoxifying, stopping bleeding and relieving pain, and the medicinal value of treating large-area burns and scalds by water and fire by external application. As the Choerospondias axillaris has stronger adaptability, li Dong and the like find that the Choerospondias axillaris can be mainly distributed in areas with higher temperature and precipitation sensitivity. She Xuemin et al found that the phenotype variation of Choerospondias axillaris in different habitats is large due to different regional climates, and that the variation of the nutritional ingredients of fruits is relatively obvious, and that there is also a large difference between the climatic period and the disease resistance. In addition, tian Hualin et al find that the seedling-stage characters of the Choerospondias axillaris have larger intra-seed variation in research on natural forest resources of the Choerospondias axillaris and research on the seedling-stage character variation of the Choerospondias axillaris, have extremely obvious differences among seed sources, have higher generalized genetic force, and are beneficial to selection of the euonymus alatus. These all indicate that the choerospondias axillaris accumulate abundant intraspecies genetic variation during the long-term evolution in subtropical and tropical regions of China. Genetic variation is an important material basis for species genetic improvement, and the research on the genetic diversity of the Choerospondias axillaris is less reported at present, and related researches on molecular genetics comprise Yang Chunxia et al, which analyze the Choerospondias axillaris transcriptome based on high-throughput sequencing; she Jinshan et al established and optimized the Choerospondias axillaris ISSR-PCR reaction system.
Microsatellite markers, also called short tandem repeats (Short tandem repeats, STRs), simple repeat sequences (Simple sequence repeat, SSR), are composed of tandem repeat units of 1 to 6 bases, and are widely found in the genomes of eukaryotic and prokaryotic organisms. Hamada et al found many microsatellites in eukaryotes (yeast to vertebrates), and later studies of Delseny, tautz and Renz found and demonstrated the abundance of microsatellites in plants and many other eukaryotes. Since the plants are rich in AT repeats and the animals are rich in AC repeats, the parts of the repeats are different, revealing the distinction between plant and animal genomes. SSRs are distributed in coding and non-coding regions of the genome, throughout the nuclear genome. About 104-105 SSRs are arranged in the genome, the polymorphism is high, and the polymorphism is different and repeated in a microsatellite region, so that the SSR molecular marker can be easily detected by PCR, and the SSR molecular marker is a very important, practical and convenient tool. Over time, unequal exchanges between nucleotides within the organism occur, after which bases of different repetition frequencies are gradually accumulated. In different organisms, even at the same microsatellite locus, the microsatellite is mutated to present rich polymorphism due to the different number of repeated units, and the mutation does not harm the organisms, so that the mutation of the microsatellite in the organisms is long-lasting. SSR markers have the advantages of rapid mutation, higher polymorphism and stability, abundant information content, wide distribution, co-dominance and the like, and can be used as an excellent genetic marker type (Kuroda et al, 2006). At present, the technology is widely applied to the fields of population genetic diversity research and the like. The SSR molecular marker for evaluating the genetic diversity of the Choerospondias axillaris is developed and applied to the research of the genetic diversity of different groups of the Choerospondias axillaris, and has important guiding significance in the aspects of analyzing the genetic structure and genetic diversity of different groups of the species, formulating effective protection and sustainable utilization strategies of choerospondias axillaris germplasm resources and the like.
Disclosure of Invention
The invention aims to provide an SSR molecular marker combination for evaluating the genetic diversity of the Choerospondias axillaris and application thereof, so as to solve the problems of the prior art, and the SSR molecular marker combination can be used for evaluating the genetic diversity of the Choerospondias axillaris, is used for researching the genetic diversity of different groups of the Choerospondias axillaris, and has important guiding significance in analyzing the genetic structure and the genetic diversity of different groups of the species, formulating effective protection and sustainable utilization strategies of choerospondias axillaris germplasm resources and the like.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides an SSR molecular marker combination for evaluating genetic diversity of Choerospondias axillaris, which comprises molecular markers NSZ-43385, NSZ-9141, NSZ-37910, NSZ-240, NSZ-22900, NSZ-39803, NSZ-42088, NSZ-165, NSZ-26522, NSZ-39039, NSZ-41051, NSZ-1007, NSZ-36575, NSZ-37045, NSZ-34772 and NSZ-45650;
the NSZ-43385 is obtained by amplifying a primer pair shown as SEQ ID NO. 1-2;
the NSZ-9141 is obtained by amplification of a primer pair shown as SEQ ID NO. 3-4;
the NSZ-37910 is obtained by amplification of a primer pair shown as SEQ ID NO. 5-6;
the NSZ-240 is obtained by amplification of a primer pair shown as SEQ ID NO. 7-8;
the NSZ-22900 is obtained by amplification of a primer pair shown as SEQ ID NO. 9-10;
the NSZ-39803 is obtained by amplification of a primer pair shown as SEQ ID NO. 11-12;
the NSZ-42088 is obtained by amplification of a primer pair shown as SEQ ID NO. 13-14;
the NSZ-165 is obtained by amplification of a primer pair shown as SEQ ID NO. 15-16;
the NSZ-26522 is obtained by amplification of a primer pair shown as SEQ ID NO. 17-18;
the NSZ-39039 is obtained by amplification of a primer pair shown as SEQ ID NO. 19-20;
the NSZ-41051 is obtained by amplification of a primer pair shown as SEQ ID NO. 21-22;
the NSZ-1007 is obtained by amplification of a primer pair shown as SEQ ID NO. 23-24;
the NSZ-36575 is obtained by amplification of a primer pair shown as SEQ ID NO. 25-26;
the NSZ-37045 is obtained by amplification of a primer pair shown as SEQ ID NO. 27-28;
the NSZ-34772 is obtained by amplification of a primer pair shown as SEQ ID NO. 29-30;
the NSZ-45650 is obtained by amplification of a primer pair shown as SEQ ID NO. 31-32.
The invention also provides a primer combination for evaluating the genetic diversity of the Choerospondias axillaris, which comprises 16 pairs of primer pairs, wherein the nucleotide sequences of the primer pairs are shown as SEQ ID NO. 1-32.
The invention also provides application of the primer combination in preparation of a kit for evaluating genetic diversity of the Choerospondias axillaris.
The invention also provides a kit for evaluating the genetic diversity of the Choerospondias axillaris, which comprises the primer combination.
The invention also provides application of the SSR molecular marker combination, the primer combination or the kit in evaluating the genetic diversity of the Choerospondias axillaris.
The invention also provides a method for evaluating the genetic diversity of the choerospondias axillaris, which comprises the following steps:
(1) Extracting the genome DNA of the Choerospondias axillaris;
(2) PCR amplification is performed by using the primer combination;
(3) Detecting the amplified product to obtain a detection result;
(4) And (3) carrying out genetic diversity analysis, genetic differentiation analysis or genetic structure analysis on the choerospondias axillaris population by using the detection result in the step (3).
Further, in the step (2), the reaction system of the PCR amplification is: 2X Taq PCR Master Mix 12.5.5. Mu.L, 1. Mu.L of upstream primer, 1. Mu.L of downstream primer, 1. Mu.L of DNA template, and ddH 2 O 9.5μL。
Further, in step (2), the reaction procedure of the PCR amplification is: 94 ℃ for 5min;94℃30s,63℃30s,72℃ 45s,10 cycles; 94℃for 30s,55℃for 30s,72℃for 45s,20 cycles; 7min at 72 ℃.
The invention discloses the following technical effects:
the invention utilizes the result of the Choerospondias axillaris transcriptome sequencing to screen out SSR molecular marker combinations suitable for evaluating the choerospondias axillaris genetic diversity, and can be used for analyzing the genetic structure of the species among different populations and the genetic diversity of the populations. The SSR molecular marker combination provided by the invention is beneficial to analyzing and knowing the genetic variation distribution pattern of the Choerospondias axillaries, and provides a theoretical basis for the efficient utilization and protection of the Choerospondias axillaries germplasm resources.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a fluorescent label of 3 primer pairs; wherein A is FAM fluorescent label of the primer pair NSZ-37910; b is HEX fluorescent label of the primer pair NSZ-240; c is TAMRA fluorescent label of a primer pair NSZ-39039;
FIG. 2 shows the amplification results of two Choerospondias axillaris samples at 24 pairs of primers; A. in the B, the single marked Choerospondias axillaris sample is YF01 (Yifeng 01), and the double marked Choerospondias axillaris sample is YF07 (Yifeng 07); in a and B, 13: marker,1-12 and 14-25:24 pairs of primary screening primers;
FIG. 3 is a proportion of SSRs of different tandem repeat unit types in the total SSR;
FIG. 4 is a result of a cluster analysis of 28 parts of Choerospondias axillaris germplasm resources for 4 populations using 16 pairs of polymorphic SSR primers.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
In the following examples, the choerospondias axillaris material used for SSR molecular marker screening and availability evaluation was from a choerospondias axillaris germplasm resource pool of Ganzea, chongyi county, jiangxi province, comprising 28 parts of germplasm resources of 4 groups of Jiangxi (Table 1).
TABLE 1 Choerospondias seed germplasm resource details for SSR screening
Example 1
1. SSR locus mining based on Choerospondias axillaris transcriptome splicing sequence
(1) RNA sample preparation, library preparation and quality inspection
a) Extraction of total RNA sample preparation: tissue samples were collected and the samples were then assayed. And after the quality detection of the RNA meets the requirements, library preparation is carried out.
b) After enrichment of eukaryotic mRNA with polyA tail by Oligo (dT) bearing magnetic beads, the mRNA was disrupted by ultrasound.
c) The first strand of cDNA was synthesized in an M-M. Mu.LV reverse transcriptase system using fragmented mRNA as template and random oligonucleotides as primers, followed by RNaseH degradation of the RNA strand.
d) Under the DNApolymerase I system, dNTPs are used as raw materials to synthesize a cDNA second strand.
e) And (3) carrying out end repair, A tail addition and sequencing joint installation on the purified double-stranded cDNA, screening about 200bp cDNA by using AMPure XPbeans, carrying out PCR amplification, purifying a PCR product again by using the AMPure XPbeans, and screening fragment sizes by agarose gel electrophoresis to finally obtain a library.
f) After library construction, sequencing was performed with Illumina HiseqTM PE s to obtain raw data.
(2) Data quality control
When quality evaluation and splicing are carried out, data filtering is carried out on the original data before information analysis so as to reduce data analysis interference. Firstly, performing quality control on raw reads of a machine under the control of fastp, and filtering low-quality data to obtain clean reads. The filtering steps are as follows: reads containing adapter, having an N content of more than 10% and all of A bases and having a low quality (the number of bases having a mass value Q.ltoreq.20 is 50% or more of the whole read) were removed.
(3) Original sequence quality assessment and splicing
And (3) assembling the high-quality sequence by using the high-throughput sequencing original data obtained in the steps (1) and (2) and then using Trinity assembling software to finally obtain 40341 Unigenes serving as basic data of the experiment.
(4) SSR locus mining based on Choerospondias axillaris transcriptome
40341 Unigenes obtained by assembly are taken as basic data of the experiment. SSR identification is carried out on the data of the Choerospondias axillaris transcriptome by using MISA software.
(5) SSR motif repeat type, number and frequency signatures in transcriptomes
There are 5 types of SSR repeats, namely dinucleotide to hexanucleotide repeats. The statistics of the proportion of SSRs of different tandem repeat unit types in the total SSR are shown in figure 3, the SSR short tandem repeat units (the abscissa meaning: such as AAC/GTT, are formed by the 6 (by shifting and reverse complementation, the components are identical except the beginning and ending parts) SSRs of AAC, ACA, CAA, GTT, TGT, TTG, and the two most abundant SSRs have the highest occurrence frequency of dinucleotide repeats.
The repeated times of the SSR repeated units of the axillary choerospondias axillaris transcriptome are distributed between 4 and 26 times, wherein the total number of SSRs of 4 to 8 times is 4476, and the total number of the SSRs is 78.33%; secondly, the SSR is carried out for 9 to 14 times, and the total number of the SSR is 952, which accounts for 16.66 percent of the total number; there were 286 of 15 replicates and more, accounting for 5%. The frequency of dinucleotide and trinucleotide recurrence is dominant, accounting for 46.95% and 34.27% of the total SSR, respectively; the number of tetranucleotides, pentanucleotides, hexanucleotides and other repeat types was small, accounting for 11.64%, 3.64% and 3.50% of the total (see table 2), respectively.
TABLE 2 type, quantity and distribution frequency of Choerospondias axillaris SSR
SSR primer design and screening
5251 SSR sites are found out, primers are respectively designed for the identified SSR by using Primer3.0 software, the main technical parameters of the primer design are GC content 40% -60%, annealing temperature is 55-65 ℃ (optimal 60 ℃), the difference of Tm values of upstream and downstream primers is less than or equal to 2 ℃, the length of the primers is 18-24 bp, the expected amplification product length is 100-300 bp, no secondary structure or dimer exists, then the synthesis of SSR primers is carried out according to the obtained nucleotide arrangement sequence, and the primer 15753 pair is designed together. Randomly selecting 100 pairs from primers with polymorphism potential (SSR sequence length >15 bp) obtained by screening.
The pre-experiment is carried out by using the selected 100 pairs of primers, two different groups of wild jujube individuals are randomly selected, 100 pairs of SSR primers are subjected to PCR amplification according to the PCR reaction system of the following table 3, the amplified products are subjected to 2% agarose gel electrophoresis to detect the size of DNA fragments, the fragments are screened out to be clear and single, no specific amplification exists, the fragments with the size meeting the expected purpose are the primary screening qualified primer pairs, and 55 pairs of primary screening qualified primers are used (part of the primary screening qualified primers is shown in figure 2). Then 8 Choerospondias axillaris samples (YF 01, 02; CY01, 02; LN01, 02; GF01, 02) are selected, two sample DNAs are selected from four groups of Jiangxi Yichun Yifeng, ganzhou Chongyi, ganzhou Longnan and Shangshen Guangfeng, repeated screening and verification are carried out (the amplification program is shown in Table 4), 2% agarose gel electrophoresis is used for detection, and 16 pairs of effective SSR marker primer pairs with better stability, higher polymorphism and better repeatability are selected (shown in Table 5). And sending the selected amplification products meeting the requirements to a company for capillary electrophoresis detection. FIG. 1 shows fluorescent labeling of 3 primer pairs (TAMRA, FAM, HEX).
Wherein the Choerospondias axillaris DNA is extracted by CTAB method. The method comprises the following steps:
(1) placing dried leaf of Choerospondias axillaris in 2mL EP tube, placing steel ball, grinding leaf into powder (12000 rm,3 min) with plant tissue grinder, grinding, and taking out steel ball;
(2) adding 800 μl of 2×CTAB extract and 8 μl of mercaptoethanol into EP tube containing powder, mixing, placing into a water bath preheated at 65deg.C for 20min, and mixing the EP tube upside down at intervals of 6 min;
(3) taking out the EP tube, adding equal volume of chloroform isoamyl alcohol, slowly reversing upside down to fully and uniformly mix, placing the EP tube in a centrifugal machine, setting the temperature in the centrifugal machine to be 4 ℃, and centrifuging at 12000rpm for 10min;
(4) carefully sucking out the supernatant, putting the supernatant into a new 2mL EP tube, adding equal volume of chloroform isoamyl alcohol, slowly reversing the mixture to be fully and uniformly mixed, putting the mixture into a centrifuge, setting the temperature to be 4 ℃, and centrifuging the mixture at 12000rpm for 10min;
(5) carefully sucking out 500 mu L of supernatant, placing into a new 1.5mL EP tube, adding equal volume of chloroform isoamyl alcohol, gently shaking until two phases are mixed uniformly, and placing into a refrigerator at-20 ℃ for more than 3 hours;
(6) when DNA is separated out, white floccule precipitate is seen, the white floccule precipitate is placed in a centrifugal machine, the temperature is set at 4 ℃, the rotating speed is 12000rpm, and the centrifugal machine is used for 10min, and at the moment, the DNA is precipitated at the bottom of a pipe;
(7) carefully pouring out the solution in the tube, keeping the sediment at the bottom, adding 500 mu L of 75% ethanol to wash the sediment, placing the sediment in a centrifuge at a temperature of 4 ℃ and a rotating speed of 12000rpm, and centrifuging for 2min;
(8) repeating the step 7, washing the precipitate again, pouring out ethanol, airing the EP tube containing the DNA precipitate in a natural state, and adding 100 mu L of 1 xTE to dissolve the DNA;
the purity and concentration of the DNA were measured by an ultraviolet spectrophotometer. ddH for extracted DNA 2 O was diluted to 50 ng/. Mu.L and stored at-20 ℃.
TABLE 3SSR-PCR reaction System
TABLE 4SSR-PCR amplification procedure
Table 516 SSR markers for polymorphisms
The 16 pairs of polymorphism SSR primers shown in Table 5 are utilized to carry out group cluster analysis on 4 groups of 28 parts of Choerospondias axillaries materials according to different geographical positions and genetic distance pairs of Choerospondias axillaries, and the tested materials are divided into 2 major classes at a genetic distance of 0.15 (FIG. 4). Class 1 contains 21 sample materials, and at a genetic distance threshold of 0.17, the class is divided into 2 subclasses, with the two populations being grouped into one for Yichunyfeng and Ganzhou Chongyi and the other for Ganzhou Longnan. In addition, group 2 contained 7 sample materials, a group of Shangroufeng choerospondias axillaries.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (8)
1. An SSR molecular marker combination for evaluating genetic diversity of Choerospondias axillaris, which is characterized by comprising molecular markers NSZ-43385, NSZ-9141, NSZ-37910, NSZ-240, NSZ-22900, NSZ-39803, NSZ-42088, NSZ-165, NSZ-26522, NSZ-39039, NSZ-41051, NSZ-1007, NSZ-36575, NSZ-37045, NSZ-34772 and NSZ-45650;
the NSZ-43385 is obtained by amplifying a primer pair shown as SEQ ID NO. 1-2;
the NSZ-9141 is obtained by amplification of a primer pair shown as SEQ ID NO. 3-4;
the NSZ-37910 is obtained by amplification of a primer pair shown as SEQ ID NO. 5-6;
the NSZ-240 is obtained by amplification of a primer pair shown as SEQ ID NO. 7-8;
the NSZ-22900 is obtained by amplification of a primer pair shown as SEQ ID NO. 9-10;
the NSZ-39803 is obtained by amplification of a primer pair shown as SEQ ID NO. 11-12;
the NSZ-42088 is obtained by amplification of a primer pair shown as SEQ ID NO. 13-14;
the NSZ-165 is obtained by amplification of a primer pair shown as SEQ ID NO. 15-16;
the NSZ-26522 is obtained by amplification of a primer pair shown as SEQ ID NO. 17-18;
the NSZ-39039 is obtained by amplification of a primer pair shown as SEQ ID NO. 19-20;
the NSZ-41051 is obtained by amplification of a primer pair shown as SEQ ID NO. 21-22;
the NSZ-1007 is obtained by amplification of a primer pair shown as SEQ ID NO. 23-24;
the NSZ-36575 is obtained by amplification of a primer pair shown as SEQ ID NO. 25-26;
the NSZ-37045 is obtained by amplification of a primer pair shown as SEQ ID NO. 27-28;
the NSZ-34772 is obtained by amplification of a primer pair shown as SEQ ID NO. 29-30;
the NSZ-45650 is obtained by amplification of a primer pair shown as SEQ ID NO. 31-32.
2. The primer combination for evaluating the genetic diversity of the Choerospondias axillaris is characterized by comprising 16 pairs of primer pairs, wherein the nucleotide sequences of the primer pairs are shown as SEQ ID NO. 1-32.
3. Use of the primer combination of claim 2 in the preparation of a kit for evaluating genetic diversity of choerospondias axillaris.
4. A kit for evaluating genetic diversity of choerospondias axillaris, comprising the primer combination of claim 2.
5. Use of the SSR molecular marker combination according to claim 1, the primer combination according to claim 2 or the kit according to claim 4 in evaluating genetic diversity of choerospondias axillaris.
6. The method for evaluating the genetic diversity of the choerospondias axillaris is characterized by comprising the following steps of:
(1) Extracting the genome DNA of the Choerospondias axillaris;
(2) Performing PCR amplification using the primer combination of claim 2;
(3) Detecting the amplified product to obtain a detection result;
(4) And (3) carrying out analysis of the genetic diversity, genetic differentiation or genetic structure of the axillary choerospondias axillaries by using the detection result of the step (3).
7. The method according to claim 6, wherein in the step (2), the reaction system of the PCR amplification is: 2X Taq PCR Master Mix 12.5.5. Mu.L, 1. Mu.L of upstream primer, 1. Mu.L of downstream primer, 1. Mu.L of DNA template, and ddH 2 O9.5μL。
8. The method of claim 6, wherein in step (2), the reaction procedure for PCR amplification is: 94 ℃ for 5min;94℃30s,63℃30s,72℃45s,10 cycles; 94℃for 30s,55℃for 30s,72℃for 45s,20 cycles; 7min at 72 ℃.
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| CN116790790A (en) * | 2023-06-26 | 2023-09-22 | 河北省农林科学院经济作物研究所 | Wild jujube SSR molecular marker development method and application thereof in wild jujube germplasm resource identification |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116790790A (en) * | 2023-06-26 | 2023-09-22 | 河北省农林科学院经济作物研究所 | Wild jujube SSR molecular marker development method and application thereof in wild jujube germplasm resource identification |
| CN116790790B (en) * | 2023-06-26 | 2023-12-29 | 河北省农林科学院经济作物研究所 | Wild jujube SSR molecular marker development method and application thereof in wild jujube germplasm resource identification |
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