China rose fragrance whole genome SNP molecular marker combination and application thereof

Abstract

The invention discloses a China rose fragrance whole genome SNP molecular marker combination and application thereof, 2068 SNP markers significantly related to fragrance are screened out by analyzing 358 parts of China rose re-sequencing materials and flower fragrance phenotype data, and a whole genome selection model is constructed. The method can accurately predict the fragrance character of the China rose, improve the breeding efficiency and accuracy, shorten the breeding period and reduce the cost. Meanwhile, a new direction is provided for China rose fragrance improvement and breeding, the method has reference value for character improvement of other horticultural crops, has efficient and stable capture efficiency, and can be used for subsequent sample detection and evaluation.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of molecular breeding, and particularly relates to a full-genome SNP molecular marker combination for Rosa chinensis flower fragrance and application thereof. BACKGROUND

[0002] In the field of Rosa chinensis breeding, traditional breeding methods mainly rely on phenotypic selection and multiple generations of screening and cultivation. This method not only takes a long time and has low efficiency, but also is easily affected by environmental factors, making it difficult to quickly obtain plants with ideal traits. In order to overcome the limitations of traditional breeding methods, molecular marker-assisted breeding technology has emerged and developed rapidly in recent decades. Among them, single nucleotide polymorphism (SNP) as the third generation of molecular markers has gradually become the mainstream molecular marker in current molecular genetics research and breeding application due to its significant advantages such as dense distribution, high genetic stability, easy automation analysis, and close linkage with the genome.

[0003] SNP refers to a polymorphic site in the genome caused by the variation of a single nucleotide. This variation can be in the form of substitution, insertion or deletion. In plant genomics research, SNP markers are widely used in gene mapping, genetic map construction, molecular marker-assisted selection, and whole-genome association analysis. For example, in the research of major crops such as corn and rice, a large number of SNP markers have been developed and applied to gene mapping and molecular breeding practices, significantly improving breeding efficiency and accuracy.

[0004] Compared with traditional molecular markers such as restriction fragment length polymorphism (RFLP) and simple sequence repeat (SSR), SNP markers have higher density and better genome coverage. Due to its high-density distribution in the genome, SNP markers can more accurately locate the gene sites related to the target traits. In addition, the detection of SNP markers can be realized through high-throughput genotyping technologies such as chip technology and sequencing technology, which not only improves the detection efficiency but also reduces the detection cost. For example, using SNP chips can simultaneously detect thousands of SNP sites in a short time, providing strong technical support for large-scale genetic analysis and breeding practice.

[0005] Although SNP markers have been widely applied in many plants, their application in the improvement of Rosa chinensis flower fragrance is still in its infancy. The genetic background of Rosa chinensis flower fragrance is complex and is affected by various environmental factors, making it difficult for traditional breeding methods to accurately predict and select plants with excellent flower fragrance traits. Therefore, it is of great significance and broad application prospects to introduce SNP marker technology into the improvement of Rosa chinensis flower fragrance.

[0006] Through high-throughput SNP typing technology, researchers can quickly and accurately identify SNP sites associated with rose flower fragrance traits, thereby achieving accurate prediction and genetic improvement of flower fragrance traits. This molecular marker-based breeding method can significantly improve breeding efficiency, shorten breeding cycle, and reduce breeding cost. In addition, combined with advanced breeding techniques such as whole genome selection, breeding programs can be further optimized to rapidly improve rose flower fragrance traits.

[0007] At present, there is a lack of a high-efficiency and accurate rose flower fragrance whole genome SNP molecular marker combination and its application. SUMMARY

[0008] To solve the problems of the prior art, the purpose of the present application is to provide a rose flower fragrance whole genome SNP molecular marker combination and its application.

[0009] To achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0010] In a first aspect, the present application provides a rose flower fragrance whole genome SNP molecular marker combination.

[0011] In a second aspect, the present application provides a rose flower fragrance prediction probe.

[0012] In a third aspect, the present application provides a rose flower fragrance prediction chip.

[0013] In a fourth aspect, the present application provides an application of the SNP molecular marker combination, probe or chip in rose flower fragrance prediction.

[0014] In a fifth aspect, the present application provides a method for predicting rose flower fragrance based on the SNP molecular marker combination.

[0015] In a sixth aspect, the present application provides an application of the SNP site combination, probe, chip or prediction method in rose molecular breeding.

[0016] In a seventh aspect, the present application provides a rose molecular breeding method based on SNP markers.

[0017] The first aspect of the present application provides a rose flower fragrance whole genome SNP molecular marker combination, which includes 2068 SNP sites distributed on 14 chromosomes of rose; the specific information of the SNP site is represented in the structural form of chromosome number_ physical position_ allelic genotype, and the SNP site contains a dominant allele significantly associated with rose flower fragrance, and the specific site information is shown in Table 1 of the specification.

[0018] Further, the SNP site is determined by whole genome association analysis of 358 rose resequencing materials, multiple software and model association screening.

[0019] The second aspect of the application provides a probe for predicting the fragrance of Chinese rose, and the screening principle of the probe includes: being designed by a SNP site combination, being capable of specifically binding to the SNP site, being used for capturing a target DNA region, realizing detection of a specific SNP site, being associated with the SNP site combination in sequence information, and meeting various screening principles of probe design, including no Indel marker within 50 bp upstream and downstream of the SNP site, only one copy number of each 100 bp sequence in the whole genome, a GC content of 40-60%, no short fragment repeat sequence in the sequence, and no N base.

[0020] The third aspect of the application provides a prediction chip for the fragrance of Chinese rose, the prediction chip containing a probe for detecting the 2068 SNP sites, by detecting the genotype of the SNP site in the genomic DNA of the Chinese rose sample to be detected, and using a pre-constructed whole genome selection model, the fragrance of Chinese rose is accurately predicted.

[0021] The fourth aspect of the application provides an application of a SNP molecular marker combination, a probe or a chip in the prediction of the fragrance of Chinese rose, by extracting the genomic DNA of Chinese rose plants, and using the SNP site combination, the probe or the chip for detection.

[0022] The fifth aspect of the application provides a method for predicting the fragrance of Chinese rose based on a SNP molecular marker combination, including the following steps:

[0023] (1) extracting the genomic DNA of the Chinese rose plant to be detected;

[0024] (2) constructing a library for the genomic DNA;

[0025] (3) using a probe to hybridize and capture the DNA fragments in the library, and enriching the DNA fragments containing the target SNP site;

[0026] (4) performing high-throughput sequencing on the captured DNA fragments;

[0027] (5) comparing and analyzing the sequencing data with a Chinese rose reference genome to determine the genotype of the SNP site;

[0028] (6) inputting the genotype data of the SNP site combination into a whole genome selection model to obtain the prediction result of the fragrance of the Chinese rose to be detected.

[0029] Further, the prediction accuracy of Bayes Ridge Regression (BRR) in the model is the highest, and the overall prediction accuracy is greater than 0.657.

[0030] The sixth aspect of the present application provides an application of the SNP site combination, the probe, the chip or the prediction method in the molecular breeding of Chinese rose, which is used for early, accurate and efficient prediction and screening of the flower fragrance trait of the breeding material in the breeding process of Chinese rose, so as to shorten the breeding cycle, improve the selection efficiency, and cultivate new varieties of Chinese rose with excellent flower fragrance trait.

[0031] Further, the whole genome selection model constructed based on the SNP site combination can be used to predict the flower fragrance of Chinese rose, and the prediction accuracy is greater than 0.657.

[0032] The seventh aspect of the present application provides a SNP marker-based molecular breeding method of Chinese rose, which comprises the following steps: applying the SNP site combination, the prediction probe or the prediction chip to the breeding process of Chinese rose varieties, detecting the genotype at the seed or seedling stage to predict the flower fragrance trait, and screening out plants with ideal flower fragrance, so as to accelerate the cultivation of new varieties of Chinese rose with less or even no thorns, improve the ornamental value and production efficiency of Chinese rose, and meet the market demand.

[0033] Beneficial effects: the present application can shorten the breeding cycle, improve the breeding efficiency, reduce the cost, improve the prediction accuracy, and has good application prospect, can accelerate the cultivation of new varieties of Chinese rose with less or even no thorns, and meet the market demand.

[0034] Compared with the prior art, the present application has the following advantages:

[0035] (1) improving the breeding efficiency and accuracy: the traditional Chinese rose breeding method is time-consuming and low in efficiency, and it needs to be screened and cultivated for multiple generations to obtain plants with ideal traits. The present application uses SNP site combination, probe and chip technologies to quickly locate the gene sites related to the flower fragrance trait of Chinese rose, realizes accurate selection, significantly improves the breeding efficiency and accuracy, and speeds up the breeding process of excellent varieties. Through the constructed whole genome selection model, the DNA of the seed or seedling can be directly detected and the future flower fragrance trait can be predicted, without going through the tedious field planting and screening process, further shortening the breeding cycle and reducing the breeding cost.

[0036] (2) reducing the research cost: compared with the traditional prediction model based on the combination of phenotype data and genomic information, the SNP site combination and other technologies used in the present application can more accurately select plants with target traits, reduce unnecessary experimental materials and operations, and thus reduce the research cost.

[0037] (3) providing a new research direction and technical support: the present application provides a new research direction for the improvement and breeding of Chinese rose flower fragrance, i.e. using SNP site combination, probe and chip and other molecular marker assisted breeding technologies. At the same time, it also provides important reference value and practical guidance for the trait improvement of other horticultural crops, and has wide application prospect.

[0038] (4) Realize the accurate prediction of the floral fragrance trait: through whole genome association analysis of 358 Rosa chinensis re-sequencing materials and floral fragrance phenotype data, 2068 SNP markers significantly related to fragrance are screened, and a whole genome selection model with excellent prediction results is successfully constructed, thereby providing a solid theoretical basis and data support for the accurate prediction and genetic improvement of the floral fragrance of Rosa chinensis.

[0039] (5) Application advantages of the liquid breeding chip: the SNP sites of the present application can be used to prepare a liquid breeding chip, which has a high-efficiency technical means and can more conveniently and quickly detect and analyze a large number of Rosa chinensis samples, thereby further improving the efficiency and accuracy of breeding work.

[0040] (6) Stable capture efficiency: the results of evaluating the liquid probe capture efficiency of 12 Rosa chinensis varieties in Example 3 show that the detection rate of 2068 SNPs ranges from 97.62% to 98.81%, the coverage depth is 391.08-531.36X, the overall capture efficiency is good, and the probe and chip of the present application have good stability and reliability, and can be used for SNP detection and whole genome selection evaluation of subsequent other samples. BRIEF DESCRIPTION OF DRAWINGS

[0041] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed in the embodiments or prior art description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0042] Figure 1 A whole chromosome marker distribution map of the floral fragrance of Rosa chinensis provided by the present application.

[0043] Figure 2 An allele frequency map of the floral fragrance of Rosa chinensis provided by the present application.

[0044] Figure 3 A deletion rate map of the floral fragrance of Rosa chinensis provided by the present application.

[0045] Figure 4 A heterozygosity map of the floral fragrance of Rosa chinensis provided by the present application.

[0046] Figure 5 A whole genome selection evaluation result map of the floral fragrance of Rosa chinensis provided by the present application. DETAILED DESCRIPTION

[0047] Clearly, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative effort are within the protection scope of the present application.

[0048] In the present application, the term "and / or" describes the association relationship of the associated objects, which means that there can be three relationships, for example, A and / or B can represent the following three cases: A exists alone, A and B exist together, and B exists alone. Wherein A and B can be singular or plural. The character " / " generally represents an "or" relationship between the associated objects before and after it.

[0049] In the present application, "one or more" means one or more, and "multiple" means two or more. "One or more of the following items" or the like means any combination of the items, including any combination of single item or multiple items. For example, "one or more of a, b, or c", or "one or more of a, b, and c" can represent a, b, c, a-b (i.e. a and b), a-c, b-c, or a-b-c, wherein a, b, and c can be single or multiple.

[0050] It should be understood that in various embodiments of the present application, the size of the sequence number of the above-mentioned processes does not mean the order of execution, and part or all of the steps can be executed in parallel or in sequence. The execution order of each process should be determined according to its function and inherent logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0051] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments, and are not intended to limit the present application. The singular forms "a", "said" and "the" used in the embodiments of the present application and the appended claims are also intended to include the plural forms, unless the context clearly indicates otherwise.

[0052] The weight of the related components mentioned in the specification of the embodiments of the present application can not only refer to the specific content of each component, but also represent the weight ratio relationship between each component. Therefore, as long as the content of the related components in the specification of the embodiments of the present application is enlarged or reduced in proportion, it is within the scope disclosed in the specification of the embodiments of the present application. Specifically, the mass mentioned in the specification of the embodiments of the present application can be μg, mg, g, kg and other mass units commonly known in the chemical field.

[0053] The first aspect of the embodiment of the present application provides a full-genome SNP molecular marker combination of Chinese rose flower fragrance, the SNP molecular marker combination comprises 2068 SNP sites distributed on 14 chromosomes of Chinese rose; the specific information of the SNP site is represented in the structural form of chromosome number_ physical position_ allelic genotype, and the SNP site comprises a dominant allele positively significantly associated with the flower fragrance of Chinese rose, and the specific site information is shown in Table 1.

[0054] The SNP site is determined through full-genome association analysis on 358 Chinese rose resequencing materials, and multiple software and model association screening.

[0055] The second aspect of the embodiment of the present application provides a prediction probe of Chinese rose flower fragrance, and the screening principle of the probe comprises the following: the probe is designed from the SNP site combination, can be specifically combined with the SNP site, is used for capturing a target DNA region, realizes detection of a specific SNP site, the sequence information of the probe is associated with the SNP site combination, and meets the screening principles of probe design, including that there is no Indel marker within 50 bp of the upstream and downstream of the SNP site, there is only one copy number of the sequence of 100 bp of the upstream and downstream in the whole genome, the GC content is 40-60 %, there is no short fragment repeat sequence in the sequence, and there is no N base.

[0056] The third aspect of the embodiment of the present application provides a prediction chip of Chinese rose flower fragrance, the prediction chip comprises probes for detecting the 2068 SNP sites, by detecting the genotype of the SNP site in the genomic DNA of the Chinese rose sample to be detected, and using a pre-constructed full-genome selection model, the flower fragrance of Chinese rose is accurately predicted.

[0057] The fourth aspect of the embodiment of the present application provides application of the SNP molecular marker combination, the probe or the chip in prediction of Chinese rose flower fragrance, by extracting the genomic DNA of Chinese rose plants, and using the SNP site combination, the probe or the chip for detection.

[0058] The fifth aspect of the embodiment of the present application provides a method for predicting Chinese rose flower fragrance based on the SNP molecular marker combination, comprising the following steps:

[0059] (1) extracting the genomic DNA of the Chinese rose plant to be detected;

[0060] (2) constructing a library for the genomic DNA;

[0061] (3) using a probe to hybridize and capture the DNA fragments in the library, and enriching the DNA fragments containing the target SNP site;

[0062] (4) performing high-throughput sequencing on the captured DNA fragments;

[0063] (5) The sequencing data is compared with the Rosa reference genome to determine the genotypes of the SNP sites;

[0064] (6) The genotype data of the SNP site combination is input into the whole genome selection model to obtain the prediction result of the flower fragrance trait of the Rosa to be tested.

[0065] In some embodiments, the prediction accuracy of Bayes Ridge Regression (BRR) in the model is the highest, and the overall prediction accuracy is greater than 0.657.

[0066] The sixth aspect of the present application provides an application of the SNP site combination, the probe, the chip or the prediction method in Rosa molecular breeding, which is used for early, accurate and efficient prediction and screening of the flower fragrance trait of the breeding material in the breeding process of Rosa, so as to shorten the breeding cycle, improve the selection efficiency, and cultivate new varieties of Rosa with excellent flower fragrance trait.

[0067] In some embodiments, the whole genome selection model constructed based on the SNP site combination can predict the flower fragrance of Rosa, and the prediction accuracy is greater than 0.657.

[0068] The seventh aspect of the present application provides a Rosa molecular breeding method based on SNP markers, comprising the following steps: applying the SNP site combination, the prediction probe or the prediction chip in the breeding process of Rosa varieties, detecting the genotype at the seed or seedling stage to predict the flower fragrance trait, so as to screen out plants with ideal flower fragrance, accelerate the cultivation of new varieties of Rosa with little or no thorns, improve the ornamental value and production efficiency of Rosa, and meet the market demand.

[0069] Embodiment 1

[0070] The SNP molecular marker combination of the Rosa flower fragrance whole genome of the present application comprises 2068 SNP sites distributed on 14 chromosomes of Rosa; the specific information of the SNP site is represented in the structural form of chromosome number_ physical position_ allelic genotype, the SNP site comprises a dominant allele significantly associated with the flower fragrance of Rosa, and the specific site information is shown in Table 1 of the specification.

[0071] Table 1

[0072] Chr1A_6619422_C / T, Chr1A_6619424_T / A, Chr1A_6619431_G / T, Chr1A_8969426_G / A, Chr1A_39585429_C / G, Chr1A_40684584_A / T, Chr1A_40684615_C / T, Chr1A_42187304_A / G, Chr1A_44766135_T / C, Chr1A_44766156_A / G, Chr1A_60723100_A / G, Chr1A_60723104_T / C, Chr1B_2750504_C / T, Chr1B_20377391_G / A, Chr1B_24469104_T / C, Chr1B_28491686_C / G, Chr1B_28848995_T / C, Chr1B_29526911_T / C, Chr1B_29550729_C / T, Chr1B_29550892_A / G, Chr1B_49088165_G / A, Chr1B_49088176_A / C, Chr2A_9774768_T / C, Chr2A_18364806_G / T, Chr2A_22175943_A / G, Chr2A_22624142_T / C, Chr2A_34842359_C / T, Chr2A_52792202_C / A, Chr2A_52818956_G / A, Chr2A_69705064_C / T, Chr2A_69705080_T / C, Chr2A_79599199_T / C, Chr2A_79599253_C / A, Chr2A_79599260_C / G, Chr2A_79599384_C / T, Chr2A_79599385_A / G, Chr2A_80646510_G / A, Chr2A_80646529_G / A, Chr2A_80646665_A / C, Chr2A_80647930_C / T, Chr2A_80648643_T / C, Chr2A_80648842_A / G, Chr2A_80648896_A / G, Chr2A_80648962_A / G, Chr2A_80649027_A / T, Chr2A_80731734_G / A, Chr2A_80736724_G / A, Chr2A_80736796_G / A, Chr2A_80737801_G / A, Chr2A_80737966_G / A, Chr2A_80738093_C / A, Chr2A_80738101_A / T, Chr2A_80738149_G / A,Chr2A_80738309_A / G, Chr2A_80738406_T / G, Chr2A_80738433_G / A, Chr2A_80739095_T / C, Chr2A_80739347_C / T, Chr2A_81284208_G / A, Chr2A_81286821_G / A, Chr2A_81509094_C / A, Chr2A_81587536_A / G, Chr2A_81904155_T / A, Chr2A_81904181_A / G, Chr2A_81956522_T / C, Chr2A_81966972_C / G, Chr2A_81976467_A / G, Chr2A_81979087_G / A, Chr2A_81979126_T / A, Chr2A_82007341_A / C, Chr2A_82117042_A / T, Chr2A_82117137_G / C, Chr2A_82127423_C / T, Chr2A_82128665_T / C, Chr2A_82129274_T / G, Chr2A_82134639_A / C, Chr2A_82134760_A / C, Chr2A_82134950_G / A, Chr2A_82135049_T / C, Chr2A_82135142_C / G, Chr2A_82135291_A / G, Chr2A_82136234_G / A, Chr2A_82203239_A / T, Chr2A_82210893_T / G, Chr2A_82214193_C / T, Chr2A_82214238_T / A, Chr2A_82217843_A / G, Chr2A_82238755_A / C, Chr2A_82239798_A / G, Chr2A_82239858_A / T, Chr2A_82239895_G / T, Chr2A_82252463_G / A, Chr2A_82335213_A / C, Chr2A_82335215_C / A, Chr2A_82336062_A / G, Chr2A_82336076_G / A, Chr2A_82336083_C / G, Chr2A_82336130_C / A, Chr2A_82336136_T / C, Chr2A_82336243_G / A, Chr2A_82356614_G / T, Chr2A_82539042_C / T, Chr2A_82539130_G / A, Chr2A_82559829_G / A, Chr2A_82559836_G / A,Chr2A_82559879_C / G, Chr2A_82559896_G / A, Chr2A_82623185_T / C, Chr2A_82799735_C / T, Chr2A_82809730_G / A, Chr2A_82968323_G / T, Chr2A_82968527_A / G, Chr2A_82971956_G / A, Chr2A_82972045_T / C, Chr2A_82972139_A / G, Chr2A_82972826_G / A, Chr2A_83040821_C / T, Chr2A_83290598_G / T, Chr2A_83300130_C / T, Chr2A_83300131_C / G, Chr2A_83313086_A / G, Chr2A_83369579_A / T, Chr2A_83369643_A / C, Chr2A_83369665_C / A, Chr2A_83369675_A / G, Chr2A_83369682_A / G, Chr2A_83381217_T / A, Chr2A_83381243_A / T, Chr2A_83381262_C / T, Chr2A_83381305_G / T, Chr2A_83381360_T / G, Chr2A_83381391_A / G, Chr2A_83381433_C / T, Chr2A_83384549_C / T, Chr2A_83384600_C / T, Chr2A_83384622_T / C, Chr2A_83384630_G / A, Chr2A_83384670_A / G, Chr2A_83384943_C / T, Chr2A_83384966_C / A, Chr2A_83384983_T / C, Chr2A_83385071_C / T, Chr2A_83385093_C / A, Chr2A_83385112_A / T, Chr2A_83385158_G / A, Chr2A_83385260_C / T, Chr2A_83385287_C / T, Chr2A_83385318_A / G, Chr2A_83385506_A / C, Chr2A_83385608_C / T, Chr2A_83385907_C / T, Chr2A_83385936_C / A, Chr2A_83386004_C / A, Chr2A_83386632_A / C, Chr2A_83386685_C / A, Chr2A_83386733_T / C, Chr2A_83386739_G / A,Chr2A_83386867_T / C, Chr2A_83386885_T / C, Chr2A_83387181_T / C, Chr2A_83387246_C / T, Chr2A_83387330_C / G, Chr2A_83387342_A / G, Chr2A_83387525_C / A, Chr2A_83387642_C / A, Chr2A_83387653_T / C, Chr2A_83387737_G / A, Chr2A_83387782_G / T, Chr2A_83387801_C / A, Chr2A_83387834_G / A, Chr2A_83387868_A / T, Chr2A_83387913_T / A, Chr2A_83387929_G / C, Chr2A_83387933_T / C, Chr2A_83388043_G / A, Chr2A_83388854_A / G, Chr2A_83388863_A / G, Chr2A_83388951_T / C, Chr2A_83388982_C / T, Chr2A_83389000_G / T, Chr2A_83389036_G / T, Chr2A_83389044_A / T, Chr2A_83389045_A / C, Chr2A_83389047_A / T, Chr2A_83389172_A / T, Chr2A_83389183_C / T, Chr2A_83390436_G / A, Chr2A_83390507_G / A, Chr2A_83390635_G / A, Chr2A_83390647_G / C, Chr2A_83390652_A / G, Chr2A_83390686_G / A, Chr2A_83390757_C / G, Chr2A_83390759_G / A, Chr2A_83390793_T / C, Chr2A_83390794_G / A, Chr2A_83390804_T / C, Chr2A_83391009_A / G, Chr2A_83391010_C / T, Chr2A_83391020_C / G, Chr2A_83391051_T / C, Chr2A_83391053_A / G, Chr2A_83391062_C / G, Chr2A_83391117_A / T, Chr2A_83391133_C / T, Chr2A_83391139_C / T, Chr2A_83391165_T / C, Chr2A_83391200_C / T, Chr2A_83391207_A / T,Chr2A_83391234_G / A, Chr2A_83391324_A / G, Chr2A_83391336_T / C, Chr2A_83391345_A / G, Chr2A_83391404_C / A, Chr2A_83432692_A / G, Chr2A_83453034_G / T, Chr2A_83465985_C / T, Chr2A_83466034_T / A, Chr2A_83468934_C / T, Chr2A_83510615_T / A, Chr2A_83510799_A / G, Chr2A_83510816_T / C, Chr2A_83510865_G / A, Chr2A_83526008_C / A, Chr2A_83527119_T / C, Chr2A_83527146_G / T, Chr2A_83528060_G / A, Chr2A_83528762_G / A, Chr2A_83530442_C / G, Chr2A_83530463_G / A, Chr2A_83530477_A / G, Chr2A_83532688_A / C, Chr2A_83532710_A / G, Chr2A_83532724_A / T, Chr2A_83582145_G / T, Chr2A_83582146_A / T, Chr2A_83584709_C / T, Chr2A_83584711_C / G, Chr2A_83593046_T / A, Chr2A_83593197_A / G, Chr2A_83593213_G / A, Chr2A_83613921_A / G, Chr2A_83637618_A / G, Chr2A_83690356_C / G, Chr2A_83690656_G / A, Chr2A_83736064_T / A, Chr2A_83749535_G / A, Chr2A_83749982_C / T, Chr2A_83750042_G / T, Chr2A_83750415_T / C, Chr2A_83757394_A / G, Chr2A_83757771_A / G, Chr2A_83769743_G / A, Chr2A_83823997_T / C, Chr2A_83824076_T / C, Chr2A_83824080_A / T, Chr2A_83824147_C / A, Chr2A_83824157_A / G, Chr2A_83824171_G / T, Chr2A_83824266_T / G, Chr2A_83824396_G / T,Chr2A_83834943_G / C, Chr2A_83835330_A / G, Chr2A_83835500_T / G, Chr2A_83835669_G / A, Chr2A_83856991_T / C, Chr2A_83858407_T / G, Chr2A_83884577_T / G, Chr2A_83884587_A / G, Chr2A_83884615_G / A, Chr2A_83884622_T / C, Chr2A_83884629_C / T, Chr2A_83958702_G / A, Chr2A_83980746_A / G, Chr2A_83989236_T / C, Chr2A_83989350_T / G, Chr2A_83989635_A / G, Chr2A_83989680_A / G, Chr2A_83989780_A / G, Chr2A_83990042_A / G, Chr2A_83990488_C / G, Chr2A_83990529_T / C, Chr2A_83991773_C / A, Chr2A_83992266_T / C, Chr2A_83992299_C / A, Chr2A_83993989_A / G, Chr2A_83994142_A / G, Chr2A_83994252_G / A, Chr2A_83999341_A / G, Chr2A_83999349_T / C, Chr2A_83999429_T / A, Chr2A_83999492_T / G, Chr2A_83999568_A / G, Chr2A_84191314_G / A, Chr2A_84191328_G / A, Chr2A_84191411_T / A, Chr2A_84191500_A / G, Chr2A_84191583_G / A, Chr2A_84192074_A / G, Chr2A_84192202_A / G, Chr2A_84192315_T / C, Chr2A_84192345_T / A, Chr2A_84254780_G / A, Chr2A_84254783_G / T, Chr2A_84336174_G / A, Chr2A_84336558_G / C, Chr2A_84336677_C / G, Chr2A_84343154_A / G, Chr2A_84343214_T / C, Chr2A_84343224_T / C, Chr2A_84343270_T / C, Chr2A_84343277_G / A, Chr2A_84343323_G / C,Chr2A_84375533_T / C, Chr2A_84375583_C / T, Chr2A_84375631_G / A, Chr2A_84375639_A / T, Chr2A_84375761_G / A, Chr2A_84375800_A / G, Chr2A_84376990_A / G, Chr2A_84377015_G / C, Chr2A_84377267_T / C, Chr2A_84377547_T / C, Chr2A_84377572_G / A, Chr2A_84377593_C / A, Chr2A_84377963_C / T, Chr2A_84377995_G / C, Chr2A_84378113_A / G, Chr2A_84378139_T / G, Chr2A_84378181_G / T, Chr2A_84378186_C / G, Chr2A_84386573_T / C, Chr2A_84386597_C / T, Chr2A_84392871_G / A, Chr2A_84417635_G / A, Chr2A_84433084_T / G, Chr2A_84433184_T / G, Chr2A_84433294_C / T, Chr2A_84539089_T / C, Chr2A_84539097_A / T, Chr2A_84539121_T / A, Chr2A_84540113_A / T, Chr2A_84540126_C / T, Chr2A_84540128_A / C, Chr2A_84540129_T / C, Chr2A_84540133_C / T, Chr2A_84540140_A / G, Chr2A_84540169_A / G, Chr2A_84540177_G / A, Chr2A_84540179_A / G, Chr2A_84540183_A / G, Chr2A_84540184_G / A, Chr2A_84540242_G / A, Chr2A_84580413_C / T, Chr2A_84583521_A / T, Chr2A_84583532_A / T, Chr2A_84583573_A / T, Chr2A_84598502_T / G, Chr2A_84607002_C / T, Chr2A_84607038_T / A, Chr2A_84607112_C / T, Chr2A_84613295_A / G, Chr2A_84613308_A / G, Chr2A_84613467_T / C, Chr2A_84613476_G / A,Chr2A_84621001_T / A, Chr2A_84621020_A / G, Chr2A_84658073_A / T, Chr2A_84679833_G / A, Chr2A_84680003_T / G, Chr2A_84680133_A / T, Chr2A_84680842_T / C, Chr2A_84681603_A / C, Chr2A_84709318_C / T, Chr2A_84709322_G / A, Chr2A_84845582_G / C, Chr2A_84845723_G / C, Chr2A_84846063_A / C, Chr2A_84857236_A / C, Chr2A_84857238_T / C, Chr2A_84857311_C / T, Chr2A_84862412_C / T, Chr2A_84863964_A / G, Chr2A_84863995_A / G, Chr2A_84864380_G / A, Chr2A_84864381_T / C, Chr2A_84864382_G / A, Chr2A_84865076_C / T, Chr2A_84865096_G / C, Chr2A_84865169_A / T, Chr2A_84865221_A / T, Chr2A_84865266_A / C, Chr2A_84865274_A / C, Chr2A_84865334_T / C, Chr2A_84865340_C / G, Chr2A_84865355_G / T, Chr2A_84866129_A / C, Chr2A_84866284_C / T, Chr2A_84866350_A / G, Chr2A_84866372_T / A, Chr2A_84866424_G / A, Chr2A_84866470_T / G, Chr2A_84866514_A / G, Chr2A_84866552_A / C, Chr2A_84866557_T / A, Chr2A_84866894_T / A, Chr2A_84867161_A / T, Chr2A_84871481_G / C, Chr2A_84871842_A / T, Chr2A_84871940_A / C, Chr2A_84872277_T / C, Chr2A_84872280_C / T, Chr2A_84872347_C / A, Chr2A_84872348_G / A, Chr2A_84872396_T / G, Chr2A_84872525_C / A, Chr2A_84872590_A / C,Chr2A_84874254_T / G, Chr2A_84874358_T / C, Chr2A_84874370_G / A, Chr2A_84874400_A / G, Chr2A_84874402_C / T, Chr2A_84874422_T / C, Chr2A_84874427_G / T, Chr2A_84874455_T / G, Chr2A_84874556_T / C, Chr2A_84874560_C / A, Chr2A_84874607_G / A, Chr2A_84874803_G / T, Chr2A_84874824_T / C, Chr2A_84874875_A / G, Chr2A_85043015_G / A, Chr2A_85043071_C / T, Chr2A_85043074_T / C, Chr2A_85043117_A / G, Chr2A_85043380_G / A, Chr2A_85043605_A / T, Chr2A_85124741_C / T, Chr2A_85131480_C / G, Chr2A_85153258_G / A, Chr2A_85202983_T / C, Chr2A_85202991_G / A, Chr2A_85203015_C / T, Chr2A_85203016_T / G, Chr2A_85203034_T / G, Chr2A_85203035_A / T, Chr2A_85203044_G / T, Chr2A_85203061_T / C, Chr2A_85203081_C / G, Chr2A_85203101_G / A, Chr2A_85203108_G / A, Chr2A_85203114_G / A, Chr2A_85243027_T / C, Chr2A_85243033_T / C, Chr2A_85243115_T / A, Chr2A_85243407_A / G, Chr2A_85273338_C / T, Chr2A_85274637_A / G, Chr2A_85311885_C / T, Chr2A_85341239_G / A, Chr2A_85341276_T / A, Chr2A_85373046_T / G, Chr2A_85400381_T / C, Chr2A_85401494_T / G, Chr2A_85401537_C / T, Chr2A_85410101_G / A, Chr2A_85451555_T / G, Chr2A_85451557_C / A, Chr2A_85451560_T / G,Chr2A_85451564_C / G, Chr2A_85451588_G / T, Chr2A_85471725_G / A, Chr2A_85471810_T / A, Chr2A_85484933_A / G, Chr2A_85484970_G / A, Chr2A_85484981_C / A, Chr2A_85484993_C / T, Chr2A_85485033_A / G, Chr2A_85504186_A / G, Chr2A_85519249_T / G, Chr2A_85519506_C / T, Chr2A_85519516_C / T, Chr2A_85521404_T / C, Chr2A_85533159_T / C, Chr2A_85533421_C / T, Chr2A_85533548_C / T, Chr2A_85548380_C / T, Chr2A_85551475_C / A, Chr2A_85551699_G / A, Chr2A_85551868_A / T, Chr2A_85553081_A / G, Chr2A_85553131_A / G, Chr2A_85553273_G / C, Chr2A_85553290_T / G, Chr2A_85555302_C / T, Chr2A_85659996_G / C, Chr2A_85660181_T / C, Chr2A_85660265_G / A, Chr2A_85661764_T / G, Chr2A_85661858_T / C, Chr2A_85688693_A / T, Chr2A_85688861_A / G, Chr2A_85688874_A / G, Chr2A_85691341_C / A, Chr2A_85693484_T / C, Chr2A_85693545_G / T, Chr2A_85694695_T / A, Chr2A_85695032_G / C, Chr2A_85697375_G / A, Chr2A_85697463_T / A, Chr2A_85697498_A / C, Chr2A_85699705_A / G, Chr2A_85699709_A / G, Chr2A_85699741_G / C, Chr2A_85699856_A / G, Chr2A_85699860_C / G, Chr2A_85699867_A / G, Chr2A_85699871_A / C, Chr2A_85699879_C / A, Chr2A_85699883_A / G, Chr2A_85699888_T / C,Chr2A_85699890_T / G, Chr2A_85699894_G / C, Chr2A_85699900_T / C, Chr2A_85699914_T / C, Chr2A_85699923_A / G, Chr2A_85699924_T / C, Chr2A_85700008_A / G, Chr2A_85700014_A / G, Chr2A_85700083_C / T, Chr2A_85700106_C / T, Chr2A_85700288_A / G, Chr2A_85700336_G / T, Chr2A_85700338_G / T, Chr2A_85701866_A / G, Chr2A_85701877_G / C, Chr2A_85701948_G / A, Chr2A_85702242_A / T, Chr2A_85702243_A / T, Chr2A_85702244_A / T, Chr2A_85702398_T / C, Chr2A_85705660_C / T, Chr2A_85705706_C / T, Chr2A_85718365_C / A, Chr2A_85718407_T / A, Chr2A_85723530_A / G, Chr2A_85723558_A / G, Chr2A_85723783_A / C, Chr2A_85723854_T / C, Chr2A_85723862_G / A, Chr2A_85723907_A / G, Chr2A_85723979_A / G, Chr2A_85723996_A / G, Chr2A_85724023_C / T, Chr2A_85724047_C / T, Chr2A_85724865_G / C, Chr2A_85724867_T / G, Chr2A_85724988_G / A, Chr2A_85725024_A / T, Chr2A_85740874_G / T, Chr2A_85744618_G / A, Chr2A_85745008_A / G, Chr2A_85749793_C / T, Chr2A_85751586_T / G, Chr2A_85767594_T / A, Chr2A_85767615_C / G, Chr2A_85767624_G / A, Chr2A_85767804_C / T, Chr2A_85767809_C / A, Chr2A_85770759_T / G, Chr2A_85775450_T / G, Chr2A_85775466_G / C, Chr2A_85800086_G / A,Chr2A_85800869_T / C, Chr2A_85800907_A / T, Chr2A_85801114_A / T, Chr2A_85803530_A / C, Chr2A_85806241_T / G, Chr2A_85806261_C / G, Chr2A_85806369_A / C, Chr2A_85806370_G / T, Chr2A_85806406_G / A, Chr2A_85806412_C / T, Chr2A_85806420_A / G, Chr2A_85806491_C / G, Chr2A_85806550_C / A, Chr2A_85806724_C / T, Chr2A_85806748_A / G, Chr2A_85806753_T / C, Chr2A_85806818_C / T, Chr2A_85806984_C / T, Chr2A_85807058_T / A, Chr2A_85807059_C / T, Chr2A_85807066_T / G, Chr2A_85807122_G / T, Chr2A_85807289_A / G, Chr2A_85807318_A / G, Chr2A_85807424_G / A, Chr2A_85819054_A / G, Chr2A_85820853_G / C, Chr2A_85820898_G / A, Chr2A_85820914_G / A, Chr2A_85820961_G / A, Chr2A_85821033_C / T, Chr2A_85823591_A / T, Chr2A_85828883_A / G, Chr2A_85829383_C / T, Chr2A_85830370_T / G, Chr2A_85830446_C / T, Chr2A_85830478_G / A, Chr2A_85830613_G / A, Chr2A_85830623_G / T, Chr2A_85830645_T / C, Chr2A_85831573_G / A, Chr2A_85831609_T / C, Chr2A_85831619_T / C, Chr2A_85831641_T / C, Chr2A_85831961_C / G, Chr2A_85840528_C / T, Chr2A_85840544_T / A, Chr2A_85840545_T / A, Chr2A_85848277_A / G, Chr2A_85863418_G / A, Chr2A_85863473_A / C, Chr2A_85866584_T / G,Chr2A_85867398_A / G, Chr2A_85892772_C / T, Chr2A_85892821_G / T, Chr2A_85949066_C / A, Chr2A_85949106_C / G, Chr2A_85949131_A / C, Chr2A_86002218_C / T, Chr2A_86017599_C / A, Chr2A_86027521_G / A, Chr2A_86027522_A / T, Chr2A_86028594_A / G, Chr2A_86046909_T / G, Chr2A_86100705_G / A, Chr2A_86127102_T / A, Chr2A_86127378_C / T, Chr2A_86127382_T / G, Chr2A_86127390_C / T, Chr2A_86127430_G / A, Chr2A_86131831_T / A, Chr2A_86131845_G / T, Chr2A_86135726_G / A, Chr2A_86135759_C / G, Chr2A_86135851_T / A, Chr2A_86135853_T / C, Chr2A_86135942_T / A, Chr2A_86136158_T / C, Chr2A_86137410_C / T, Chr2A_86147446_G / A, Chr2A_86147512_C / T, Chr2A_86147544_T / G, Chr2A_86147631_G / A, Chr2A_86147719_G / A, Chr2A_86147780_T / C, Chr2A_86156419_A / G, Chr2A_86165443_A / T, Chr2A_86167561_T / A, Chr2A_86167620_A / T, Chr2A_86203192_G / C, Chr2A_86203230_A / G, Chr2A_86203283_T / A, Chr2A_86203300_T / G, Chr2A_86208696_C / A, Chr2A_86217601_T / C, Chr2A_86217664_A / T, Chr2A_86254514_A / T, Chr2A_86255255_C / T, Chr2A_86255256_G / A, Chr2A_86255257_A / G, Chr2A_86255465_G / A, Chr2A_86255471_T / C, Chr2A_86255521_A / C, Chr2A_86256150_A / G,Chr2A_86256216_C / T, Chr2A_86258949_G / A, Chr2A_86259026_A / G, Chr2A_86259040_C / A, Chr2A_86283199_T / C, Chr2A_86283248_A / T, Chr2A_86321303_C / G, Chr2A_86325148_C / T, Chr2A_86325156_C / T, Chr2A_86325193_C / A, Chr2A_86325243_C / T, Chr2A_86326252_T / C, Chr2A_86326255_C / G, Chr2A_86492440_A / G, Chr2A_86492450_G / C, Chr2A_86492840_G / T, Chr2A_86493641_G / A, Chr2A_86497231_A / T, Chr2A_86511146_G / A, Chr2A_86513094_A / G, Chr2A_86513138_T / A, Chr2A_86513148_T / C, Chr2A_86513886_A / T, Chr2A_86560606_A / T, Chr2A_86586299_A / G, Chr2A_86632921_G / A, Chr2A_86649582_A / C, Chr2A_86843761_C / T, Chr2A_86920704_C / T, Chr2A_86920818_T / C, Chr2A_86923223_C / G, Chr2A_86923583_C / T, Chr2A_86924333_G / A, Chr2A_86924352_C / T, Chr2A_86925997_A / G, Chr2A_86926179_A / G, Chr2A_87031351_T / C, Chr2A_87148787_T / C, Chr2A_87300092_G / A, Chr2A_87300114_C / T, Chr2A_87322124_A / G, Chr2A_87322174_C / G, Chr2A_87326491_A / G, Chr2A_87326497_T / G, Chr2A_87326501_T / C, Chr2A_87326606_G / A, Chr2A_87326613_A / G, Chr2A_87505530_T / C, Chr2A_87550812_A / G, Chr2A_87550866_T / C, Chr2A_87559748_T / C, Chr2A_87901135_A / G,Chr2A_88151118_T / C, Chr2A_88481320_A / G, Chr2B_9783471_A / C, Chr2B_47904348_G / T, Chr2B_50415639_A / G, Chr2B_52110643_C / G, Chr2B_66452214_T / G, Chr2B_66665054_T / C, Chr2B_66812481_T / G, Chr2B_66812486_A / G, Chr2B_67283070_A / C, Chr2B_67485216_G / A, Chr2B_67550498_T / G, Chr2B_67550523_G / A, Chr2B_68437768_T / C, Chr2B_68509668_G / A, Chr2B_68509673_G / A, Chr2B_68613351_A / G, Chr2B_70940452_G / C, Chr2B_77397344_A / T, Chr2B_77397402_C / T, Chr2B_77397404_G / C, Chr2B_77397418_T / C, Chr2B_77397447_C / A, Chr2B_77397630_A / T, Chr2B_77399460_A / G, Chr2B_77399496_G / A, Chr2B_77399601_C / T, Chr2B_77399602_C / T, Chr2B_77399661_C / T, Chr2B_77400013_G / A, Chr2B_77400496_T / C, Chr2B_77400786_C / G, Chr2B_77401420_G / A, Chr2B_77401515_G / A, Chr2B_77401527_A / T, Chr2B_77401678_A / C, Chr2B_77401750_A / G, Chr2B_77401755_C / G, Chr2B_77401763_C / T, Chr2B_77401891_G / C, Chr2B_77534327_C / T, Chr2B_77612698_G / A, Chr2B_78817386_C / T, Chr2B_78919898_C / G, Chr2B_78934452_G / A, Chr2B_78934456_T / C, Chr2B_78935004_G / C, Chr2B_78935108_C / T, Chr2B_78935113_T / A, Chr2B_78935114_G / A, Chr2B_78935126_C / T,Chr2B_78935127_C / T, Chr2B_78935154_G / A, Chr2B_78935180_G / C, Chr2B_78968110_C / T, Chr2B_79173842_T / A, Chr2B_81346635_C / G, Chr2B_81405335_G / A, Chr2B_81409028_G / A, Chr2B_81420341_G / A, Chr2B_81424386_G / A, Chr2B_81424855_A / C, Chr2B_81424934_G / A, Chr2B_81425980_C / T, Chr2B_81426291_G / A, Chr2B_81427698_A / G, Chr2B_81427824_A / C, Chr2B_81427977_T / C, Chr2B_81427996_G / A, Chr2B_81431907_A / T, Chr2B_81693264_C / T, Chr2B_81750260_G / T, Chr2B_82045413_T / C, Chr2B_82046060_T / C, Chr2B_82057366_G / T, Chr2B_82061061_A / G, Chr2B_82061084_A / G, Chr2B_82061197_C / G, Chr2B_82061976_T / C, Chr2B_82062544_T / C, Chr2B_82062632_T / C, Chr2B_82062718_C / A, Chr2B_82062769_A / G, Chr2B_82062809_A / T, Chr2B_82062811_A / T, Chr2B_82062838_G / A, Chr2B_82062840_A / T, Chr2B_82062842_A / T, Chr2B_82062860_G / A, Chr2B_82062920_C / T, Chr2B_82062940_C / A, Chr2B_82062983_T / C, Chr2B_82063446_A / T, Chr2B_82063452_A / C, Chr2B_82063545_A / G, Chr2B_82063549_A / C, Chr2B_82063560_A / T, Chr2B_82063575_G / T, Chr2B_82063712_C / A, Chr2B_82064033_A / T, Chr2B_82064164_T / C, Chr2B_82064498_T / C, Chr2B_82065661_C / T,Chr2B_82065760_A / G, Chr2B_82066659_C / T, Chr2B_82068566_A / C, Chr2B_82068569_C / T, Chr2B_82068584_A / T, Chr2B_82068761_G / A, Chr2B_82080553_C / G, Chr2B_82081539_T / C, Chr2B_82081601_T / A, Chr2B_82081664_T / C, Chr2B_82081735_C / A, Chr2B_82081784_C / A, Chr2B_82082561_G / A, Chr2B_82235810_A / G, Chr2B_82292899_G / T, Chr2B_82366432_A / T, Chr2B_82423511_A / G, Chr2B_82513744_C / T, Chr2B_82639197_C / A, Chr2B_82639275_T / A, Chr2B_82667799_T / C, Chr2B_82667809_T / C, Chr2B_82667942_C / T, Chr2B_82668041_G / A, Chr2B_82668254_A / T, Chr2B_82668329_T / C, Chr2B_82668366_T / A, Chr2B_82668367_G / A, Chr2B_82785632_A / T, Chr2B_82785659_C / G, Chr2B_82785763_A / G, Chr2B_82785817_T / C, Chr2B_82838279_C / T, Chr2B_82842591_A / G, Chr2B_82842847_C / T, Chr2B_82843909_C / A, Chr2B_82848062_A / G, Chr2B_82857369_G / A, Chr2B_82857443_T / C, Chr2B_82885064_A / T, Chr2B_82917288_G / A, Chr2B_82972183_A / G, Chr2B_83066332_C / T, Chr2B_83077493_G / T, Chr2B_83085821_A / T, Chr2B_83588724_G / T, Chr2B_83999554_G / A, Chr2B_84026885_A / G, Chr2B_84026904_A / G, Chr2B_84026906_T / G, Chr2B_84026934_T / G, Chr2B_84131073_T / C,Chr2B_84131203_G / C, Chr2B_84198272_T / C, Chr2B_84242183_A / G, Chr2B_84550851_G / A, Chr2B_84692720_C / T, Chr3A_384459_C / T, Chr3A_384533_A / T, Chr3A_384537_G / A, Chr3A_384567_G / A, Chr3A_384591_C / G, Chr3A_384593_A / G, Chr3A_389124_C / T, Chr3A_546387_T / C, Chr3A_960306_T / C, Chr3A_960315_G / A, Chr3A_962070_T / C, Chr3A_1361911_A / T, Chr3A_1378120_A / G, Chr3A_1648836_A / T, Chr3A_2047314_C / T, Chr3A_2051591_C / T, Chr3A_2213771_A / T, Chr3A_2248919_G / C, Chr3A_2252546_C / T, Chr3A_2252548_C / T, Chr3A_2252709_G / A, Chr3A_2253009_G / T, Chr3A_21707136_A / T, Chr3A_21707149_T / G, Chr3A_28867477_T / G, Chr3A_28867491_A / G, Chr3A_28867494_T / C, Chr3A_28867495_T / C, Chr3A_28867504_G / A, Chr3A_28867513_T / G, Chr3A_29178076_A / G, Chr3A_31845695_G / T, Chr3A_32030496_C / T, Chr3A_33417523_C / T, Chr3B_590794_G / A, Chr3B_1611931_C / T, Chr3B_17108889_T / G, Chr3B_31165061_G / A, Chr4A_483435_A / T, Chr4A_5269663_G / A, Chr4A_5269664_T / C, Chr4A_9702976_G / A, Chr4A_23077322_C / T, Chr4A_23077323_T / C, Chr4A_23747279_A / G, Chr4A_23791066_A / G, Chr4A_23791279_C / T, Chr4A_23822010_A / G, Chr4A_25564226_A / G,Chr4A_28034371_T / C, Chr4A_34244524_C / T, Chr4A_34431207_G / T, Chr4A_34433585_A / G, Chr4A_34434511_G / A, Chr4A_34439148_A / G, Chr4A_35046252_A / G, Chr4A_35046257_C / T, Chr4A_35195515_T / C, Chr4A_35266496_G / A, Chr4A_36703974_G / A, Chr4A_36875621_G / A, Chr4A_45415189_A / G, Chr4A_46489233_C / G, Chr4A_48260206_C / G, Chr4A_48260220_C / T, Chr4A_48462319_G / A, Chr4A_48462327_A / G, Chr4A_48462861_G / A, Chr4A_48560262_A / T, Chr4A_48881712_C / T, Chr4A_49086324_G / A, Chr4A_49156507_T / C, Chr4A_49235946_G / A, Chr4A_49320759_G / A, Chr4A_49320760_A / T, Chr4A_49320765_G / C, Chr4A_49320993_T / C, Chr4A_49321004_C / G, Chr4A_49336378_C / T, Chr4A_49336583_G / A, Chr4A_49346031_T / C, Chr4A_49347935_G / C, Chr4A_49348000_A / G, Chr4A_49348008_T / C, Chr4A_49719723_T / C, Chr4A_49720024_T / A, Chr4A_49794249_G / A, Chr4A_49851486_G / A, Chr4A_49851691_C / T, Chr4A_49855435_G / C, Chr4A_49894667_G / A, Chr4A_50035551_C / G, Chr4A_50035608_T / C, Chr4A_50035611_C / T, Chr4A_50035803_C / T, Chr4A_50209873_T / C, Chr4A_50340081_C / T, Chr4A_50456795_T / A, Chr4A_50511432_A / T, Chr4A_50741671_G / A, Chr4A_58894474_T / C,Chr4A_59476339_C / T, Chr4A_59511287_T / C, Chr4A_59516499_C / T, Chr4A_59612404_A / G, Chr4A_59655187_T / G, Chr4A_59655419_G / T, Chr4A_59656113_T / C, Chr4A_59700640_G / A, Chr4A_59712667_A / C, Chr4A_59712683_T / C, Chr4A_59967125_G / A, Chr4A_60008636_T / G, Chr4A_60096058_T / A, Chr4A_60105671_A / C, Chr4A_60139205_C / T, Chr4A_60144855_C / T, Chr4A_60162772_T / C, Chr4A_60163430_C / A, Chr4A_60170215_C / T, Chr4A_60170267_T / A, Chr4A_60170310_A / G, Chr4A_60172682_C / T, Chr4A_60172686_C / T, Chr4A_60172745_C / T, Chr4A_60177230_A / G, Chr4A_60178310_G / A, Chr4A_60178488_T / A, Chr4A_60180305_G / T, Chr4A_60181571_C / T, Chr4A_60181842_C / T, Chr4A_60185276_G / C, Chr4A_60237466_A / G, Chr4A_60246864_G / T, Chr4A_60246907_G / A, Chr4A_60247062_C / T, Chr4A_60247095_T / C, Chr4A_60247215_A / G, Chr4A_60247360_G / A, Chr4A_60357476_C / T, Chr4A_60368070_C / A, Chr4A_60368185_T / C, Chr4A_60368210_A / G, Chr4A_60491271_G / A, Chr4A_60491278_C / G, Chr4A_60491302_A / T, Chr4A_60531934_T / A, Chr4A_60531935_G / A, Chr4A_60532145_G / A, Chr4A_60552542_C / T, Chr4A_60579749_G / A, Chr4A_60579817_A / T, Chr4A_60579900_C / T,Chr4A_60579905_A / T, Chr4A_60580383_C / T, Chr4A_60580388_T / C, Chr4A_60580391_T / G, Chr4A_60626383_T / C, Chr4A_60626452_C / A, Chr4A_60632048_A / G, Chr4A_60632066_C / T, Chr4A_60632451_A / G, Chr4A_60632500_A / C, Chr4A_60632909_T / C, Chr4A_60637149_A / G, Chr4A_60637294_G / A, Chr4A_60639149_A / G, Chr4A_60639159_G / T, Chr4A_60663046_G / A, Chr4A_60663068_G / A, Chr4A_60663070_G / C, Chr4A_60663086_G / A, Chr4A_60663244_C / T, Chr4A_60670476_T / A, Chr4A_60670483_A / T, Chr4A_60670511_C / T, Chr4A_60672961_C / A, Chr4A_60674376_G / A, Chr4A_60674385_T / C, Chr4A_60678056_A / G, Chr4A_60678057_G / A, Chr4A_60680926_A / G, Chr4A_60680933_A / T, Chr4A_60680950_G / A, Chr4A_60718876_C / G, Chr4A_60718888_C / T, Chr4A_60718891_T / C, Chr4A_60722158_G / A, Chr4A_60745018_C / T, Chr4A_60745121_T / A, Chr4A_60855499_A / G, Chr4A_60855507_C / T, Chr4A_60856054_C / T, Chr4A_60856158_C / T, Chr4A_60856439_C / T, Chr4A_60856460_G / A, Chr4A_60856472_T / C, Chr4A_60856474_G / A, Chr4A_60858649_G / A, Chr4A_60858709_C / T, Chr4A_60858712_C / T, Chr4A_60864296_A / G, Chr4A_60864297_T / C, Chr4A_60864301_C / T, Chr4A_60864302_T / C,Chr4A_60864332_C / T, Chr4A_60864363_T / C, Chr4A_60864377_G / A, Chr4A_60864417_G / A, Chr4A_60864450_G / A, Chr4A_60865012_G / C, Chr4A_60865022_G / A, Chr4A_60865034_T / C, Chr4A_60865059_T / C, Chr4A_60865063_C / T, Chr4A_60865069_C / T, Chr4A_60865204_G / A, Chr4A_60865213_G / A, Chr4A_60865248_C / G, Chr4A_60865255_C / T, Chr4A_60865328_C / T, Chr4A_60865339_A / G, Chr4A_60865342_C / T, Chr4A_60865343_A / G, Chr4A_60865427_A / C, Chr4A_60865437_C / G, Chr4A_60865481_C / T, Chr4A_60865541_A / G, Chr4A_60865612_G / A, Chr4A_60875576_T / C, Chr4A_60903005_T / C, Chr4A_60903016_A / C, Chr4A_60917442_T / C, Chr4A_60917518_C / A, Chr4A_60917554_A / G, Chr4A_60917913_T / A, Chr4A_60917962_T / C, Chr4A_60920747_G / T, Chr4A_60920753_G / A, Chr4A_60924707_G / T, Chr4A_60972313_A / G, Chr4A_60972339_A / G, Chr4A_60972351_A / T, Chr4A_61037402_A / C, Chr4A_61053159_G / A, Chr4A_61053191_A / G, Chr4A_61065713_C / T, Chr4A_61101007_G / A, Chr4A_61101025_C / T, Chr4A_61183356_C / G, Chr4A_61183565_A / G, Chr4A_61185229_G / C, Chr4A_61185296_C / T, Chr4A_61185314_A / T, Chr4A_61185329_C / T, Chr4A_61213023_C / T, Chr4A_61213040_A / G,Chr4A_61215676_C / T, Chr4A_61215694_G / A, Chr4A_61215714_A / C, Chr4A_61215753_A / G, Chr4A_61215762_A / T, Chr4A_61227634_A / G, Chr4A_61236149_A / G, Chr4A_61236224_A / G, Chr4A_61289109_T / A, Chr4A_61297602_G / C, Chr4A_61297606_G / A, Chr4A_61297713_T / C, Chr4A_61342494_G / A, Chr4A_61346678_C / A, Chr4A_61346689_C / T, Chr4A_61346692_C / T, Chr4A_61347101_G / A, Chr4A_61347167_C / A, Chr4A_61347272_C / A, Chr4A_61347331_T / C, Chr4A_61397899_A / T, Chr4A_61397901_A / T, Chr4A_61398540_T / A, Chr4A_61398560_G / C, Chr4A_61409892_G / A, Chr4A_61409906_T / C, Chr4A_61410012_T / C, Chr4A_61410016_C / T, Chr4A_61445653_G / A, Chr4A_61445685_T / A, Chr4A_61445900_G / A, Chr4A_61445934_G / A, Chr4A_61470384_A / G, Chr4A_61502490_G / T, Chr4A_61518460_T / A, Chr4A_61518462_G / A, Chr4A_61518467_T / G, Chr4A_61540803_G / C, Chr4A_61584464_T / A, Chr4A_61601734_C / A, Chr4A_61603693_C / A, Chr4A_61603700_G / A, Chr4A_61604472_T / A, Chr4A_61801864_G / A, Chr4A_61829577_A / T, Chr4A_61857035_A / G, Chr4A_61857045_G / T, Chr4A_61863222_A / G, Chr4A_61868015_C / G, Chr4A_61877132_A / G, Chr4A_61877134_G / A, Chr4A_61877234_A / G,Chr4A_61877271_T / G, Chr4A_61877297_A / G, Chr4A_61877305_C / A, Chr4A_61877602_T / C, Chr4A_62077894_C / T, Chr4A_62235044_G / T, Chr4A_62235059_C / G, Chr4A_62235079_G / A, Chr4A_62235104_T / C, Chr4A_62235108_T / G, Chr4A_62656004_G / A, Chr4A_62726525_A / G, Chr4A_62741190_A / G, Chr4A_62759833_C / T, Chr4A_62759851_C / G, Chr4A_62759880_T / C, Chr4A_62827156_C / G, Chr4A_62827162_T / G, Chr4A_62837507_T / C, Chr4A_62842744_A / G, Chr4A_62915221_G / A, Chr4A_62915423_C / G, Chr4A_62929102_A / C, Chr4A_62929117_A / T, Chr4A_62942180_A / T, Chr4A_62949550_C / G, Chr4A_62954446_A / T, Chr4A_62958562_A / C, Chr4A_62958679_T / C, Chr4A_62967215_G / A, Chr4A_62967336_A / G, Chr4A_62993065_A / G, Chr4A_62993089_T / A, Chr4A_62993090_T / A, Chr4A_62993284_A / G, Chr4A_63032283_G / T, Chr4A_63032305_A / C, Chr4A_63032775_G / A, Chr4A_63032832_G / A, Chr4A_63032846_C / T, Chr4A_63036396_T / C, Chr4A_63036434_A / G, Chr4A_63039070_C / T, Chr4A_63039297_G / T, Chr4A_63039330_C / T, Chr4A_63062185_A / G, Chr4A_63062188_C / G, Chr4A_63062202_C / A, Chr4A_63062352_A / G, Chr4A_63153723_T / G, Chr4A_63154072_A / T, Chr4A_63154123_G / T,Chr4A_63154163_T / C, Chr4A_63154284_C / T, Chr4A_63155012_G / T, Chr4A_63155752_G / A, Chr4A_63156183_C / T, Chr4A_63156289_C / T, Chr4A_63156414_T / C, Chr4A_63156499_G / A, Chr4A_63157422_T / A, Chr4A_63157440_A / G, Chr4A_63157565_A / T, Chr4A_63157570_T / C, Chr4A_63157594_C / T, Chr4A_63157936_G / C, Chr4A_63157968_C / G, Chr4A_63158019_G / A, Chr4A_63158200_A / T, Chr4A_63158239_T / C, Chr4A_63158240_T / G, Chr4A_63168157_A / T, Chr4A_63168222_A / G, Chr4A_63168275_A / G, Chr4A_63168973_C / G, Chr4A_63168989_A / G, Chr4A_63169055_G / A, Chr4A_63169149_T / C, Chr4A_63185666_A / G, Chr4A_63185700_G / T, Chr4A_63185723_A / C, Chr4A_63209237_T / A, Chr4A_63222146_A / G, Chr4A_63222219_A / T, Chr4A_63222326_C / T, Chr4A_63222338_C / T, Chr4A_63227371_C / G, Chr4A_63237694_G / A, Chr4A_63238072_G / T, Chr4A_63238077_A / T, Chr4A_63238679_T / C, Chr4A_63238720_T / C, Chr4A_63238780_T / A, Chr4A_63238795_C / T, Chr4A_63238826_G / A, Chr4A_63238912_G / A, Chr4A_63238916_G / A, Chr4A_63238946_C / G, Chr4A_63242775_T / G, Chr4A_63242838_C / T, Chr4A_63257614_A / G, Chr4A_63257651_A / G, Chr4A_63257664_A / G, Chr4A_63264570_T / C,Chr4A_63264625_A / G, Chr4A_63275064_G / C, Chr4A_63283516_G / C, Chr4A_63283525_A / T, Chr4A_63286280_A / G, Chr4A_63286425_C / T, Chr4A_63286698_G / T, Chr4A_63286718_A / G, Chr4A_63286740_T / G, Chr4A_63286792_T / A, Chr4A_63288435_G / A, Chr4A_63293155_T / C, Chr4A_63303395_G / A, Chr4A_63303412_T / A, Chr4A_63303484_A / C, Chr4A_63303655_G / T, Chr4A_63307564_T / C, Chr4A_63322187_G / T, Chr4A_63322247_A / G, Chr4A_63329908_G / T, Chr4A_63336629_A / G, Chr4A_63344482_A / G, Chr4A_63365102_G / A, Chr4A_63365108_G / C, Chr4A_63365113_G / A, Chr4A_63365128_T / C, Chr4A_63365131_T / C, Chr4A_63371255_G / T, Chr4A_63371263_T / C, Chr4A_63460101_C / T, Chr4A_63472356_A / G, Chr4A_63475022_C / A, Chr4A_63475675_T / C, Chr4A_63493173_A / G, Chr4A_63530943_G / T, Chr4A_63530948_A / C, Chr4A_63530952_G / T, Chr4A_63559397_T / C, Chr4A_63559417_G / C, Chr4A_63559419_A / G, Chr4A_63559548_C / G, Chr4A_63559553_A / T, Chr4A_63559565_A / T, Chr4A_63559825_G / T, Chr4A_63559835_T / C, Chr4A_63559854_A / G, Chr4A_63566885_C / T, Chr4A_63566894_A / C, Chr4A_63566924_A / G, Chr4A_63566930_C / T, Chr4A_63566937_A / G, Chr4A_63643097_G / C,Chr4A_63643594_C / T, Chr4A_63655108_C / T, Chr4A_63655256_T / A, Chr4A_63716722_T / C, Chr4A_63716824_T / C, Chr4A_63716998_T / C, Chr4A_63717043_T / A, Chr4A_63737672_G / T, Chr4A_63737701_T / C, Chr4A_63752818_C / T, Chr4A_63753040_A / G, Chr4A_63753628_C / A, Chr4A_63753638_T / G, Chr4A_63761775_C / T, Chr4A_63761890_A / G, Chr4A_63765272_G / C, Chr4A_63765359_T / C, Chr4A_63765453_C / G, Chr4A_63765811_T / A, Chr4A_63767810_G / A, Chr4A_63767841_G / A, Chr4A_63785052_A / G, Chr4A_63785081_T / C, Chr4A_63797044_A / G, Chr4A_63816011_G / C, Chr4A_63823559_C / G, Chr4A_63861323_C / T, Chr4A_63861536_G / A, Chr4A_63861538_T / C, Chr4A_63861539_A / T, Chr4A_63861597_G / A, Chr4A_63861606_T / C, Chr4A_63908770_T / C, Chr4A_63909025_G / T, Chr4A_63909080_G / A, Chr4A_63936776_C / T, Chr4A_63943218_C / G, Chr4A_63959558_T / G, Chr4A_63970824_T / C, Chr4A_63970970_G / A, Chr4A_63970971_G / C, Chr4A_63980886_G / T, Chr4A_63994290_G / T, Chr4A_63994338_A / G, Chr4A_63994427_C / T, Chr4A_63994448_T / C, Chr4A_64013674_T / C, Chr4A_64013699_G / C, Chr4A_64020059_G / A, Chr4A_64020095_A / G, Chr4A_64034659_A / G, Chr4A_64034758_A / G,Chr4A_64034788_G / T, Chr4A_64044813_C / A, Chr4A_64047867_C / T, Chr4A_64047900_C / G, Chr4A_64047973_C / G, Chr4A_64048080_G / A, Chr4A_64048182_T / G, Chr4A_64048301_G / C, Chr4A_64060975_T / G, Chr4A_64072793_G / A, Chr4A_64078779_A / G, Chr4A_64078795_A / G, Chr4A_64078875_G / A, Chr4A_64107756_A / G, Chr4A_64107814_G / C, Chr4A_64110966_A / G, Chr4A_64123217_T / C, Chr4A_64123238_A / T, Chr4A_64123318_A / C, Chr4A_64130829_A / T, Chr4A_64130830_A / G, Chr4A_64130854_A / T, Chr4A_64144492_C / T, Chr4A_64144634_A / C, Chr4A_64144893_C / T, Chr4A_64144936_T / C, Chr4A_64145004_G / C, Chr4A_64145123_G / A, Chr4A_64145156_T / A, Chr4A_64165896_A / G, Chr4A_64165910_C / T, Chr4A_64165918_T / A, Chr4A_64165919_G / A, Chr4A_64165945_A / G, Chr4A_64182879_G / A, Chr4A_64210812_C / T, Chr4A_64249666_G / A, Chr4A_64284026_G / T, Chr4A_64322478_T / C, Chr4A_64752517_A / C, Chr4A_64754597_T / G, Chr4A_64754935_T / C, Chr4A_64873484_A / G, Chr4A_64873501_C / T, Chr4A_64978022_G / A, Chr4A_65234608_C / T, Chr4A_65469327_C / T, Chr4A_65470523_T / C, Chr4A_65470710_G / T, Chr4A_65470725_T / C, Chr4A_65470757_C / T, Chr4A_65473338_A / T,Chr4A_65474022_T / C, Chr4A_65474059_T / G, Chr4A_65543817_A / G, Chr4A_65543938_G / A, Chr4A_65558029_T / C, Chr4A_65559538_G / T, Chr4A_65660192_T / C, Chr4A_65696802_C / T, Chr4A_65696841_A / C, Chr4A_65837809_T / A, Chr4A_65837827_A / G, Chr4A_65905695_T / A, Chr4A_65905730_T / A, Chr4A_65906027_A / T, Chr4A_65911061_G / A, Chr4A_65911063_A / G, Chr4A_65924095_G / A, Chr4A_65948351_G / A, Chr4A_65948399_T / C, Chr4A_65948508_T / A, Chr4A_65948515_G / A, Chr4A_65948565_T / C, Chr4A_65948606_A / C, Chr4A_65949458_A / C, Chr4A_65949491_A / C, Chr4A_65949501_C / T, Chr4A_65949566_A / G, Chr4A_65957929_C / G, Chr4A_65958384_A / T, Chr4A_65967203_A / T, Chr4A_65967228_T / A, Chr4A_65973945_G / A, Chr4A_65973971_C / T, Chr4A_65979840_C / T, Chr4A_66079357_A / C, Chr4A_66079386_T / C, Chr4A_66079544_A / G, Chr4A_66155699_G / A, Chr4A_66163551_C / A, Chr4A_66167874_A / G, Chr4A_66167878_T / C, Chr4A_66167886_A / G, Chr4A_66169919_A / G, Chr4A_66169931_G / A, Chr4A_66170060_G / A, Chr4A_66176305_T / C, Chr4A_66176759_C / T, Chr4A_66178167_T / C, Chr4A_66178172_A / C, Chr4A_66178205_G / C, Chr4A_66178372_C / T, Chr4A_66178378_C / T,Chr4A_66178390_A / T, Chr4A_66178403_T / G, Chr4A_66178494_G / A, Chr4A_66226707_C / A, Chr4A_66226741_C / T, Chr4A_66226755_G / A, Chr4A_66232235_G / C, Chr4A_66232520_T / C, Chr4A_66235491_C / A, Chr4A_66236066_A / G, Chr4A_66236608_C / T, Chr4A_66236629_G / T, Chr4A_66237078_C / T, Chr4A_66237304_T / A, Chr4A_66253062_T / C, Chr4A_66269645_A / C, Chr4A_66273367_A / G, Chr4A_66388719_C / A, Chr4A_66388755_G / T, Chr4A_66389007_T / C, Chr4A_66389058_G / C, Chr4A_66389546_A / G, Chr4A_66389756_C / T, Chr4A_66389836_G / T, Chr4A_66389863_G / A, Chr4A_66426110_A / G, Chr4A_66480230_A / G, Chr4A_66497065_C / T, Chr4A_66497072_G / A, Chr4B_4308878_A / G, Chr4B_18528385_C / T, Chr4B_25004997_G / A, Chr4B_25005004_T / A, Chr4B_25005036_G / A, Chr4B_25005254_C / T, Chr4B_26779247_G / C, Chr4B_30290137_G / C, Chr4B_37088705_G / A, Chr4B_37498398_C / T, Chr4B_37854825_G / A, Chr4B_38038568_C / T, Chr4B_38047649_A / T, Chr4B_38047820_C / T, Chr4B_38047831_C / G, Chr4B_38047873_T / A, Chr4B_38048411_G / C, Chr4B_38118856_G / A, Chr4B_38161364_T / C, Chr4B_38466796_G / A, Chr4B_38535690_G / A, Chr4B_38535692_G / A, Chr4B_38583297_C / G,Chr4B_38678021_A / G, Chr4B_38687050_G / T, Chr4B_38687757_G / A, Chr4B_38687869_C / T, Chr4B_38688250_T / C, Chr4B_38690823_A / C, Chr4B_38730648_G / C, Chr4B_38739739_T / A, Chr4B_38826523_A / G, Chr4B_38828248_G / A, Chr4B_38976427_C / T, Chr4B_38984317_G / A, Chr4B_38984336_G / A, Chr4B_39022713_G / A, Chr4B_39033089_G / C, Chr4B_39232288_T / C, Chr4B_39663706_C / A, Chr4B_48398780_T / A, Chr4B_48434999_T / A, Chr4B_48442139_A / G, Chr4B_48444221_C / T, Chr4B_48444560_G / C, Chr4B_48447210_T / G, Chr4B_48449910_T / C, Chr4B_48474229_G / C, Chr4B_48474259_C / T, Chr4B_48475700_C / T, Chr4B_48478886_T / A, Chr4B_48479246_T / C, Chr4B_48486725_G / A, Chr4B_48487640_A / G, Chr4B_48488024_T / A, Chr4B_48488942_A / G, Chr4B_48488943_T / A, Chr4B_48489203_T / C, Chr4B_48489988_A / G, Chr4B_48490661_A / G, Chr4B_48497229_T / A, Chr4B_48529918_A / G, Chr4B_48532364_C / G, Chr4B_48536504_G / A, Chr4B_48536921_G / A, Chr4B_48536994_C / T, Chr4B_48537102_G / A, Chr4B_48537313_T / C, Chr4B_48539432_A / T, Chr4B_48540461_G / C, Chr4B_48540660_G / A, Chr4B_48540930_T / A, Chr4B_48540936_A / T, Chr4B_48541588_A / G, Chr4B_48541786_A / G,Chr4B_48541962_C / T, Chr4B_48542765_T / G, Chr4B_48553839_A / G, Chr4B_48640740_T / G, Chr4B_48641082_G / C, Chr4B_48643222_A / G, Chr4B_48651845_G / A, Chr4B_48664258_G / A, Chr4B_48664375_G / A, Chr4B_48664389_G / A, Chr4B_48720871_A / T, Chr4B_48756826_G / C, Chr4B_48776524_A / C, Chr4B_48779964_T / A, Chr4B_48791100_A / C, Chr4B_48906910_G / C, Chr4B_48977120_T / C, Chr4B_48977122_G / A, Chr4B_48977124_G / C, Chr4B_48977138_C / G, Chr4B_49080966_T / C, Chr4B_49080981_T / C, Chr4B_49485883_A / G, Chr4B_49623210_G / C, Chr4B_49738030_G / A, Chr4B_49853853_A / T, Chr4B_49915037_G / A, Chr4B_49915112_C / T, Chr4B_49915190_T / G, Chr4B_49915203_A / G, Chr4B_50170297_A / C, Chr4B_50529078_C / T, Chr4B_50529114_T / C, Chr4B_50530859_G / C, Chr4B_50531504_G / T, Chr4B_50531917_G / A, Chr4B_50533987_T / C, Chr4B_50534087_A / T, Chr4B_50538893_C / A, Chr4B_50550163_T / A, Chr4B_50582811_T / G, Chr4B_50627972_C / T, Chr4B_50701368_T / C, Chr4B_50766222_C / T, Chr4B_50766270_C / T, Chr4B_50766338_A / T, Chr4B_50766515_G / T, Chr4B_50766522_G / A, Chr4B_50766538_G / A, Chr4B_50766577_T / C, Chr4B_50766578_T / A, Chr4B_50766579_G / A,Chr4B_50766692_A / G, Chr4B_51052011_G / T, Chr4B_51054690_G / T, Chr4B_51199824_A / G, Chr4B_51225615_T / C, Chr4B_51303846_C / T, Chr4B_51326124_G / A, Chr4B_51386382_C / T, Chr4B_51411808_G / A, Chr4B_51411860_G / T, Chr4B_51412001_G / A, Chr4B_51436362_A / G, Chr4B_51455514_C / T, Chr4B_51455945_T / A, Chr4B_51455977_G / A, Chr4B_51456053_C / T, Chr4B_51456070_G / A, Chr4B_51456097_G / A, Chr4B_51481524_C / T, Chr4B_52971352_G / C, Chr4B_52997978_G / T, Chr4B_53000089_A / G, Chr4B_53035580_T / A, Chr4B_53035629_G / T, Chr4B_53035668_C / T, Chr4B_53035925_G / A, Chr4B_53035980_G / C, Chr4B_53051226_T / A, Chr4B_53051238_T / G, Chr4B_53051295_T / C, Chr4B_53051301_G / T, Chr4B_53051321_G / A, Chr4B_53100720_G / T, Chr4B_53134832_G / A, Chr4B_53134833_G / A, Chr4B_53244060_C / T, Chr4B_53270011_A / C, Chr4B_53270013_G / T, Chr4B_53270014_G / C, Chr4B_53295727_T / C, Chr4B_53327012_C / T, Chr4B_53337755_C / T, Chr4B_53360411_G / A, Chr4B_53360413_C / T, Chr4B_53363178_G / T, Chr4B_53363181_T / C, Chr4B_53364732_T / C, Chr4B_53364756_A / C, Chr4B_53364758_T / A, Chr4B_53364779_G / A, Chr4B_53364791_G / A, Chr4B_53364812_C / T,Chr4B_53364879_G / A, Chr4B_53364883_C / T, Chr4B_53526481_T / G, Chr4B_53586100_A / G, Chr4B_53586102_A / T, Chr4B_53677660_T / C, Chr4B_53689227_G / A, Chr4B_53689255_C / T, Chr4B_53735771_G / T, Chr4B_53743898_C / T, Chr4B_53743899_G / T, Chr4B_53796939_T / G, Chr4B_53825184_C / T, Chr4B_53825239_T / G, Chr4B_53827684_A / C, Chr4B_53829942_G / A, Chr4B_53829947_A / G, Chr4B_53839138_A / G, Chr4B_53839503_T / G, Chr4B_53839505_T / A, Chr4B_53848317_C / T, Chr4B_53883126_G / A, Chr4B_53892887_G / A, Chr4B_53897099_G / A, Chr4B_53897102_A / G, Chr4B_53897119_T / A, Chr4B_53897121_T / A, Chr4B_53897125_T / A, Chr4B_53897129_T / A, Chr4B_53897328_A / G, Chr4B_53968360_T / C, Chr4B_53968549_T / A, Chr4B_53968618_G / A, Chr4B_53970148_T / A, Chr4B_53970167_A / G, Chr4B_54360183_C / G, Chr4B_54657323_T / C, Chr4B_54993122_T / A, Chr4B_54993129_G / A, Chr4B_54993143_A / C, Chr4B_55179440_A / G, Chr4B_55179446_G / A, Chr4B_55233332_C / T, Chr4B_55462262_T / G, Chr4B_55484389_A / G, Chr4B_55484403_T / C, Chr4B_55494600_A / G, Chr4B_55526262_G / A, Chr4B_55613421_G / A, Chr4B_55661166_G / A, Chr4B_55789634_T / G, Chr4B_55789650_G / C,Chr4B_55789791_T / C, Chr4B_55789892_C / A, Chr4B_55886246_A / T, Chr4B_56139838_C / T, Chr5A_9935390_T / G, Chr5A_31238092_C / G, Chr5A_33138521_A / G, Chr5A_63024663_G / A, Chr5A_63074362_G / A, Chr5A_63665512_A / G, Chr5A_63668484_C / T, Chr5A_63668583_C / A, Chr5A_63668584_T / C, Chr5A_63668656_G / A, Chr5A_63668671_G / A, Chr5A_63668677_G / A, Chr5A_63668922_T / A, Chr5A_63669054_A / G, Chr5A_63669145_G / T, Chr5A_63669167_T / C, Chr5A_63669170_A / G, Chr5A_63669599_G / A, Chr5A_63669778_C / T, Chr5A_63669789_G / A, Chr5A_63669815_T / C, Chr5A_63669984_A / G, Chr5A_63670016_C / T, Chr5A_63670778_G / C, Chr5A_63670851_G / A, Chr5A_63670866_T / C, Chr5A_63670872_C / T, Chr5A_63670925_A / T, Chr5A_63671011_G / T, Chr5A_63671031_C / T, Chr5A_63671085_C / T, Chr5A_63671185_G / A, Chr5A_64706641_A / G, Chr5A_64706661_T / C, Chr5A_64707078_C / A, Chr5A_64794707_A / C, Chr5A_64857473_T / C, Chr5A_65675857_C / A, Chr5A_65675859_G / T, Chr5A_65747138_A / G, Chr5A_69338379_C / A, Chr5A_69797271_A / C, Chr5A_72930096_A / G, Chr5A_72930132_C / T, Chr5A_72930176_A / T, Chr5A_78951278_G / C, Chr5A_78951286_T / G, Chr5A_78951318_C / T,Chr5A_90979030_G / A, Chr5B_3470200_C / A, Chr5B_12687916_A / G, Chr5B_28415317_C / A, Chr5B_28415319_A / C, Chr5B_28415350_A / G, Chr5B_44079477_A / C, Chr5B_58863974_A / C, Chr5B_59531502_G / T, Chr5B_60188587_C / T, Chr5B_60188595_G / A, Chr5B_60281906_C / G, Chr5B_60282092_A / T, Chr5B_60412933_T / C, Chr5B_60441181_C / T, Chr5B_60519622_A / T, Chr5B_60519637_A / G, Chr5B_60646177_C / T, Chr5B_60801486_G / A, Chr5B_61159523_G / C, Chr5B_61159892_G / T, Chr5B_61374313_A / C, Chr5B_61452978_C / T, Chr5B_61534843_G / A, Chr5B_61661994_C / T, Chr5B_61710207_T / A, Chr5B_61710534_A / C, Chr5B_62413041_A / G, Chr5B_62450735_T / C, Chr5B_62450748_A / G, Chr5B_62514597_A / G, Chr5B_62514604_T / G, Chr5B_63072505_C / T, Chr5B_63073273_C / T, Chr5B_64066896_C / T, Chr5B_64388847_C / T, Chr5B_64484768_A / G, Chr5B_70190820_C / T, Chr5B_71508014_C / T, Chr5B_82097877_C / G, Chr5B_84032817_G / A, Chr5B_85620914_A / G, Chr5B_85924686_T / C, Chr5B_86739185_G / A, Chr6A_9851158_C / T, Chr6A_9945783_A / C, Chr6A_10116796_A / G, Chr6A_10116976_A / C, Chr6A_10117001_T / A, Chr6A_10117138_A / G, Chr6A_10117148_A / T, Chr6A_10117154_A / T,Chr6A_10117191_C / T, Chr6A_10220523_T / A, Chr6A_10254766_G / A, Chr6A_10286206_A / T, Chr6A_10622056_A / G, Chr6A_10622119_C / T, Chr6A_11819222_T / A, Chr6A_17336058_T / C, Chr6A_18601687_T / G, Chr6A_18601688_T / A, Chr6A_18601692_T / C, Chr6A_33769444_A / C, Chr6A_33769446_C / A, Chr6A_33769475_C / T, Chr6A_33769478_A / C, Chr6A_44100518_T / C, Chr6A_44100534_C / T, Chr6A_44190282_C / A, Chr6A_44195339_A / T, Chr6A_44202759_C / A, Chr6A_44700120_A / T, Chr6A_44700441_T / A, Chr6A_44896899_C / T, Chr6A_44900218_C / T, Chr6A_44965407_C / T, Chr6A_44965458_G / A, Chr6A_44965459_G / T, Chr6A_44972105_A / G, Chr6A_44972589_T / C, Chr6A_44972590_G / T, Chr6A_44972747_A / T, Chr6A_44972857_A / G, Chr6A_48079007_T / A, Chr6A_48079008_T / C, Chr6A_48245452_C / T, Chr6A_50466714_T / G, Chr6A_50577787_C / T, Chr6A_50578657_G / A, Chr6A_50598655_G / A, Chr6A_51777932_T / C, Chr6A_51951277_C / A, Chr6A_53678270_A / C, Chr6A_53851110_C / T, Chr6A_54929698_A / C, Chr6A_55632990_G / T, Chr6A_55633111_T / A, Chr6A_55942255_C / T, Chr6A_57813046_C / T, Chr6A_59688219_C / T, Chr6A_60023274_A / G, Chr6A_60082386_T / A, Chr6A_60320501_C / T,Chr6A_60535791_C / T, Chr6A_61213066_T / A, Chr6A_64286296_C / G, Chr6A_66098056_A / T, Chr6B_1504127_T / C, Chr6B_4813310_T / A, Chr6B_11164230_G / T, Chr6B_12950706_C / T, Chr6B_20092517_C / T, Chr6B_32787350_A / G, Chr6B_32794339_C / T, Chr6B_32794849_T / G, Chr6B_32796631_A / T, Chr6B_33868614_T / C, Chr6B_34016601_G / A, Chr6B_34016602_A / G, Chr6B_34019209_A / G, Chr6B_34019994_A / G, Chr6B_34019999_G / C, Chr6B_34020000_G / T, Chr6B_34063724_T / C, Chr6B_34063759_A / G, Chr6B_34445125_C / G, Chr6B_34446222_A / T, Chr6B_34449122_A / T, Chr6B_34449411_T / C, Chr6B_34449583_A / C, Chr6B_34449605_C / T, Chr6B_34449617_G / A, Chr6B_34449807_A / T, Chr6B_34449813_T / C, Chr6B_34449839_T / A, Chr6B_34451347_A / G, Chr6B_34452312_T / C, Chr6B_34452332_A / T, Chr6B_34467127_A / G, Chr6B_34467573_T / C, Chr6B_34487810_G / A, Chr6B_34488230_C / G, Chr6B_34488806_C / A, Chr6B_34488816_C / T, Chr6B_34488897_C / T, Chr6B_34530269_G / T, Chr6B_38219977_T / C, Chr6B_39700317_A / G, Chr6B_39807215_G / A, Chr6B_41309561_A / G, Chr6B_41442540_C / A, Chr6B_41442541_C / T, Chr6B_42122232_T / C, Chr6B_42656061_A / T, Chr6B_43015606_T / C,Chr6B_43554749_C / T, Chr6B_43554796_G / C, Chr6B_43617020_C / A, Chr6B_43619448_C / T, Chr6B_43621091_G / C, Chr6B_43627389_A / G, Chr6B_43628290_G / A, Chr6B_44109797_G / A, Chr6B_44109823_A / C, Chr6B_44109841_G / A, Chr6B_44339359_A / G, Chr6B_44396225_A / G, Chr6B_44509018_T / C, Chr6B_44558695_T / A, Chr6B_44559059_G / A, Chr6B_44628951_G / A, Chr6B_44630625_A / G, Chr6B_44630691_T / G, Chr6B_44631547_T / A, Chr6B_44631604_T / C, Chr6B_44636274_A / G, Chr6B_44662948_A / T, Chr6B_49334800_C / T, Chr6B_51605446_G / C, Chr6B_52693109_C / T, Chr6B_56626454_C / T, Chr6B_56626457_G / T, Chr6B_56736491_G / A, Chr6B_56804883_C / T, Chr6B_56807661_A / T, Chr6B_58887012_T / C, Chr6B_63813756_T / C, Chr6B_63813763_C / A, Chr6B_63813805_A / T, Chr6B_63813806_A / G, Chr6B_63813808_A / T, Chr6B_63813820_A / T, Chr6B_63813831_T / C, Chr6B_63813833_C / A, Chr7A_25861_G / A, Chr7A_227538_A / G, Chr7A_419620_C / T, Chr7A_461633_C / A, Chr7A_1086858_C / T, Chr7A_3228471_C / T, Chr7A_3228482_G / A, Chr7A_4939632_G / T, Chr7A_5387571_C / T, Chr7A_5387576_A / G, Chr7A_5387579_A / G, Chr7A_7708979_T / C, Chr7A_8054499_T / C, Chr7A_8171100_C / T,Chr7A_8392034_C / T, Chr7A_9354840_T / A, Chr7A_9355349_A / G, Chr7A_12223577_A / T, Chr7A_24239708_T / C, Chr7A_24466727_C / T, Chr7A_24977508_T / A, Chr7A_25122349_C / T, Chr7A_27211309_G / A, Chr7A_27412355_T / G, Chr7A_27412363_G / A, Chr7A_27412412_C / T, Chr7A_28788822_T / A, Chr7A_29902202_G / A, Chr7A_31518720_C / G, Chr7A_31518824_T / C, Chr7A_31521225_G / A, Chr7A_31522600_G / T, Chr7A_43640336_T / G, Chr7A_44569602_T / A, Chr7A_44959175_A / G, Chr7A_47335849_G / C, Chr7A_47336265_T / C, Chr7A_51094751_A / G, Chr7A_55615003_G / A, Chr7A_58528948_C / T, Chr7A_58529054_C / T, Chr7A_62875393_G / T, Chr7A_62875765_C / T, Chr7A_62875822_G / T, Chr7A_63994304_G / A, Chr7A_64047782_G / A, Chr7A_69255746_A / C, Chr7A_69255759_G / A, Chr7A_69255762_G / A, Chr7A_69255763_G / T,

[0073] The SNP site is determined by whole genome association analysis on 358 Rosa resequencing materials, and is screened by multiple software and model association.

[0074] Example 2

[0075] The flower-scented probe of the Rosa plant of the present application is designed by combining SNP sites, can specifically bind to the SNP site, is used for capturing the target DNA region, realizes the detection of the specific SNP site, the sequence information is associated with the SNP site combination, and meets the screening principles of probe design, including no Indel marker within 50bp upstream and downstream of the SNP site, only one copy number of 100bp sequence upstream and downstream in the whole genome, GC content is 40-60%, no short fragment repeat sequence in the sequence, and no N base.

[0076] Embodiment 3

[0077] The application discloses a rose fragrance prediction chip, which comprises probes for detecting 2068 SNP sites, and realizes accurate prediction of rose fragrance by detecting the genotypes of the SNP sites in the genomic DNA of a sample to be detected and using a previously constructed whole-genome selection model.

[0078] Embodiment 4

[0079] The application discloses a SNP molecular marker combination, a probe or a chip and application thereof in rose fragrance prediction, wherein genomic DNA of a rose plant is extracted and detected by using the SNP site combination, the probe or the chip.

[0080] Embodiment 5

[0081] The application discloses a method for predicting rose fragrance based on a SNP molecular marker combination, which comprises the following steps:

[0082] (1) extracting genomic DNA of a rose plant to be detected;

[0083] (2) constructing a library of the genomic DNA;

[0084] (3) using a probe to hybridize and capture DNA fragments in the library, and enriching DNA fragments containing target SNP sites;

[0085] (4) performing high-throughput sequencing on the captured DNA fragments;

[0086] (5) comparing and analyzing sequencing data with a rose reference genome to determine the genotypes of the SNP sites;

[0087] (6) inputting the genotype data of the SNP site combination into a whole-genome selection model to obtain a prediction result of the fragrance of the rose to be detected; the prediction accuracy of Bayes Ridge Regression (BRR) in the model is the highest, and the overall prediction accuracy is greater than 0.657.

[0088] Embodiment 6

[0089] The application discloses a SNP site combination, a probe, a chip or a prediction method and application thereof in rose molecular breeding, which is used for early, accurate and efficient prediction and screening of the fragrance of breeding materials in the process of rose breeding, so as to shorten the breeding cycle, improve the selection efficiency and cultivate new rose varieties with excellent fragrance.

[0090] The whole genome selection model constructed based on the SNP site combination can predict the flower fragrance of the Chinese rose, and the prediction accuracy is greater than 0.657.

[0091] Example 7

[0092] The SNP marker-based Chinese rose molecular breeding method comprises the following steps: applying the SNP site combination, the prediction probe or the prediction chip to the breeding process of the Chinese rose variety, detecting the genotype at the seed or seedling stage, predicting the flower fragrance trait, and screening out the plant with ideal flower fragrance, so as to accelerate the breeding of the Chinese rose variety with little or no thorn, improve the ornamental value and production efficiency of the Chinese rose, and meet the market demand.

[0093] Example 8

[0094] The flower fragrance whole genome association analysis of 358 Chinese rose whole genome resequencing data comprises the following processes:

[0095] (1) The genomic DNA of 358 Chinese rose tissues is extracted by using the magnetic bead method, the whole genome resequencing library is constructed, the DNA fragments meeting the sequencing requirements are screened by using the magnetic beads, the qbit is quantified, and the DNBSEQ-T7 of Huada is used for sequencing;

[0096] (2) The fq.gz format of the second-generation sequencing data obtained by sequencing in step (1) is aligned with the modern Chinese rose 'Samantha' genome v2 version genome by using the bwa software. The SNP variation data is analyzed by using the GATK software, and finally 115219112 high-quality SNP sites are determined,

[0097] (3) The SNP sites screened in step (2) are reserved, the allele frequency is greater than 0.01, and the deletion rate is less than 0.2, and 35335433 high-quality SNP sites are obtained.

[0098] (4) The flower fragrance and agronomic traits are collected. 1 represents no fragrance, 2 represents fragrance, and 3 represents strong fragrance. Three repeated data are collected, the average value and the BLUP value are calculated, and a total of 5 data are used for GWAS association analysis.

[0099] (5) Use emmax, gemma, rmvp (FARMCPU model, GLM model, MLM model), tassel (GLM model) four software six models to associate the flower fragrance agronomic traits respectively, the p-value threshold is set to 1E-4, and 268779 SNP markers are associated.

[0100] (6) The SNP screened in step (5) is evaluated by a probe. The screening principle is that there is no InDel marker within 50bp upstream and downstream of the SNP site; there is only one copy number of the sequence of 100bp upstream and downstream; the GC content is 40-60%; there is no short fragment repeat sequence in the sequence; and there is no N base in the sequence, and finally 5710 SNP sites are left.

[0101] (7) The SNP sites screened in step (6) are sorted according to the number of software and models associated and the association results, and 2068 SNP markers with the most software association and significant association values are screened. Figure 1 As shown in Figure 1 The rose thorn-shaped whole chromosome marker distribution map provided by the present application is provided.

[0102] (8) The SNP sites screened in step (7) are used for 13 whole genome selection models, including LightGBM (lgb), Random Forest (RF), SVR (svr), Lasso Regression (Lasso), Ridge Regression (Ridge), rrBLUP (rrblup), Reproducing Kernel Hilbert Space (RKHS), Bayes Ridge Regression (BRR), BayesianLasso (BL), BayesC, BayesB, BayesA and GBLUP, and the genotypes and flower fragrance agronomic traits of 348 materials are used for evaluating the prediction accuracy. The results show that the prediction accuracy of Ridge is the lowest, the prediction accuracy of Bayes Ridge Regression (BRR) is the highest, the overall prediction accuracy is greater than 0.657, and the prediction accuracy is good. As shown in Figure 5 As shown in Figure 5 The rose thorn-shaped whole genome selection evaluation result map provided by the present application is provided.

[0103] (9) The SNP sites screened in step (7) are synthesized into probe sequences for capture sequencing.

[0104] Example 9

[0105] Method for detecting Rosa chinensis using liquid phase probe of 2068 SNPs described in Example 8

[0106] Genomic DNA extraction: Rosa chinensis leaves were collected and stored with ice packs and timely transported back to the laboratory. The total DNA of Rosa chinensis was extracted by magnetic bead method.

[0107] Genomic DNA library construction: genomic DNA was fragmented (200-300 bp), the ends of the DNA fragments were repaired and connected to adapters, and the library was amplified by PCR. Probe hybridization to target region: the prepared SNP probe was hybridized to the Pre-PCR library, and the streptavidin-modified probe was hybridized to the complementary DNA library to capture the target DNA library. After washing off other library DNA, the Pre-PCR library after capture was purified, and the DNA concentration required for high-throughput sequencing was enriched by amplification, followed by high-throughput sequencing.

[0108] Hybridization conditions: streptavidin-labeled probes were hybridized to target libraries at 50°C for 16 h, and biotin-labeled magnetic beads were combined with streptavidin-labeled probes at room temperature for 30 min to enrich target libraries. After washing the non-target region library with wash buffer, elution was performed with nuclease-free water.

[0109] High-throughput sequencing: the library obtained after capture and amplification in step (4) was subjected to high-throughput sequencing using the DNBSEQ-T7 sequencing platform of Huada Zhi Zao, and the sequencing results of genomic DNA were obtained. The obtained data were subjected to basic cleaning treatment. Liquid phase probe capture rate evaluation: the cleaned sequencing data were aligned to the Rosa chinensis 'Samantha' genome v2 version reference sequence using bwa software, and the capture efficiency was evaluated.

[0110] Test Example 1

[0111] Evaluation of liquid phase probe capture efficiency for 12 Rosa chinensis varieties

[0112] Experimental materials

[0113] Twelve Rosa chinensis leaves were selected, stored in the refrigerator, and timely transported to the laboratory. The total DNA of each single plant was extracted by CTAB method. The sample numbers are shown in Table 2.

[0114] Table 2

[0115] Number Variety Number Variety Test 1 Bujie Test 2 Lü Apple Test 3 Xiushan Red Test 4 Golden Age Test 5 Blue Spirit Test 6 Powder Fan Test 7 Ruby Star Test 8 Rainbow Test 9 Powder Back Test 10 Perfume Woman Test 11 Titanic Test 12 Charlotte

[0116] (1) Capture efficiency evaluation

[0117] For the 12-month seasonal material evaluation results, the proportion of 2068 SNPs that can be detected ranges from 97.62-98.81%, with a coverage depth of 391.08-531.36X, and overall capture efficiency is good, which can be used for SNP detection and whole genome selection evaluation of subsequent other samples. The detection rate statistics of the captured sequencing samples are shown in Table 3:

[0118] This test focuses on analyzing the characteristics of different sample detection indicators. In sample selection, 12 samples such as Buji, Xiushanhong, and Blue Fairy were carefully selected, which are widely sourced and typical, and can fully represent the diversity within the research scope. The test uses professional high-throughput sequencing technology to conduct deep detection on each sample to obtain the coverage depth and detection rate data of the sample. During the detection process, the standardized operation process is strictly followed, the error is reduced through repeated detection, and the quality control sample is used to calibrate the detection results to ensure the accuracy and reliability of the data. After the detection is completed, the raw data is processed and counted using bioinformatics analysis tools, and finally the coverage depth and detection rate values corresponding to each sample are obtained.

[0119] Table 3 presents the coverage depth and detection rate detection results of 12 samples. The table header includes "Sample_ID", "Coverage Depth", and "Detection Rate (%)". The data categories are clearly defined. Among them, the "Titanic" sample has the highest coverage depth of 510.36, indicating that it is the most fully detected in the detection process; the "Powder Fan" sample has the lowest coverage depth of only 348.08. In terms of detection rate, "Blue Fairy" ranks first with 98.78%, and "Xiushan Red" has a relatively low detection rate of 97.12%. Overall, although the coverage depth and detection rate values of each sample are different, the detection rates are all at a high level. These data can provide important reference for subsequent research on sample characteristics, screening of high-quality samples, or optimization of detection technology, and help to further explore the biological or other scientific significance behind the samples.

[0120] Table 3

[0121] Sample ID Coverage Depth Detection Rate (%) Bujie 426.52 98.27 Xiushan Red 446.64 97.12 Blue Spirit 383.08 98.78 Ruby Star 489.58 98.46 Powder Back 447.16 98.37 Titanic 510.36 97.93 Lü Apple 458.16 98.37 Golden Age 437.1 98.83 Powder Fan 348.08 97.37 Rainbow 457.56 98.74 Perfume Woman 491.58 98.33 Charlotte 489.62 98.19

[0122] The above shows and describes the basic principles, main features and advantages of the present application. Those skilled in the art should understand that the present application is not limited to the above examples, and the above examples and descriptions in the specification are only to illustrate the principles of the present application. Without departing from the spirit and scope of the present application, various changes and improvements can be made to the present application, and the scope of protection of the present application is defined by the appended claims, the specification and their equivalents.

Claims

1. A genome-wide SNP molecular marker combination for rose fragrance, characterized by: The SNP molecular marker combination includes 2068 SNP sites distributed on the 14 chromosomes of roses; the specific information of the SNP sites is represented by the structure of chromosome number_physical position_allele type, and the SNP sites contain dominant alleles that are positively and significantly associated with the fragrance of roses. The specific site information is shown in Table 1 of the specification.

2. The genome-wide SNP molecular marker combination for rose fragrance according to claim 1, characterized in that: The SNP sites were determined by performing genome-wide association analysis on 358 rose resequencing materials and screening through multiple software and model associations.

3. A rose fragrance prediction probe, characterized by: The screening principles of the probe include: being designed from the SNP site combination of claim 1, being able to specifically bind to the SNP site, being used to capture the target DNA region, and realizing the detection of a specific SNP site, and its sequence information being associated with the SNP site combination, and complying with various screening principles of probe design, including no Indel markers within 50 bp upstream and downstream of the SNP site, only one copy number in the entire genome of each 100 bp upstream and downstream sequence, a GC content of 40-60%, no short segment repeat sequences in the sequence, and no N bases.

4. A rose fragrance prediction chip, characterized by: The prediction chip includes probes for detecting the 2068 SNP sites. By detecting the genotype of the SNP sites in the genomic DNA of the rose sample to be tested and using a pre-constructed whole-genome selection model, accurate prediction of the rose fragrance traits is achieved.

5. Use of the SNP molecular marker combination according to claim 1, the probe according to claim 3, or the chip according to claim 4 in rose fragrance prediction, characterized in that: The genomic DNA of the rose plant is extracted and detected using the SNP site combination, probe or chip.

6. A method for predicting rose fragrance based on the SNP molecular marker combination according to claim 1, characterized in that The steps include: (1) Extracting genomic DNA from the rose plants to be tested; (2) constructing a library of the genomic DNA; (3) using the probe described in claim 3 to perform hybridization capture on the DNA fragments in the library to enrich the DNA fragments containing the target SNP site; (4) performing high-throughput sequencing on the captured DNA fragments; (5) comparing the sequenced data with the rose reference genome to determine the genotype of the SNP site; (6) Inputting the genotype data of the SNP site combination described in claim 1 into the whole genome selection model described in claim 4 to obtain the prediction results of the rose fragrance trait to be tested.

7. The method for predicting rose fragrance using a combination of SNP molecular markers according to claim 6, wherein: Among the models, Bayes Ridge Regression (BRR) has the highest prediction accuracy, with an overall prediction accuracy greater than 0.

657.

8. Use of the SNP locus combination, probe, chip or prediction method according to claim 1 in rose molecular breeding, characterized in that: It is used to make early, accurate and efficient prediction and screening of the floral fragrance traits of breeding materials during the rose breeding process, thereby shortening the breeding cycle, improving selection efficiency and cultivating new rose varieties with excellent floral fragrance traits.

9. Use of the SNP locus combination, probe, chip or prediction method according to claim 8 in rose molecular breeding, characterized in that: The whole genome selection model constructed based on the SNP site combination described in claim 1 can predict the floral fragrance of roses with a prediction accuracy greater than 0.

657.

10. A rose molecular breeding method based on SNP markers, characterized in that The method comprises the following steps: applying the SNP site combination described in claim 1, the prediction probe described in claim 3 or the prediction chip described in claim 4 to the breeding process of rose varieties, detecting the genotype at the seed or seedling stage, predicting the floral fragrance traits, thereby screening out plants with ideal floral fragrance, accelerating the breeding of new rose varieties with less floral fragrance or even no prickles, improving the ornamental value and production efficiency of roses, and meeting market demand.

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