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Keywords = multiple nucleotide polymorphism (MNP)

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15 pages, 2547 KiB  
Article
Availability Evaluation and Application of MNP (Multiple Nucleotide Polymorphism) Markers in Variety Identification of Chrysanthemum
by Yanfang Liu, Qin Zhao, Tiantian Li, Cailing Teng, Hai Peng, Zongze Yao, Zhiwei Fang, Junfei Zhou, Xiaohong Yang, Juxiang Qiao, Jin Mao, Zhiyong Guan and Qiang Hu
Horticulturae 2024, 10(8), 845; https://doi.org/10.3390/horticulturae10080845 - 9 Aug 2024
Viewed by 758
Abstract
In China, PBR (Plant Breeder’s Right) applications of chrysanthemum rank first among all of the applications of ornamental crops in China due to the plant’s significant ornamental, edible, and medicinal values. However, issues of variety infringement and disputes have become increasingly prominent, and [...] Read more.
In China, PBR (Plant Breeder’s Right) applications of chrysanthemum rank first among all of the applications of ornamental crops in China due to the plant’s significant ornamental, edible, and medicinal values. However, issues of variety infringement and disputes have become increasingly prominent, and traditional molecular markers are difficult to use due to the high heterozygosity and complex ploidy of chrysanthemum. Our study explored the availability of MNP (Multiple Nucleotide Polymorphism) markers in this regard. In total, 30 representative varieties of five types were selected for the screening of MNPs, and another 136 varieties were selected for validation of the screened MNPs. Based on ddRAD-seq (Double Digest Restriction site-associated DNA sequencing) of the 30 varieties, 26,147 SNPs were screened for genetic analysis,and 487 MNPs were screened with a length from 139 to 274 bp, an average of 6.6 SNPs individually, and a repeatability rate of 99.73%. Among the 487 MNPs, 473 MNP markers were found to cover all 27 chromosomes of chrysanthemum. Performance of our MNPs in the 136 varieties was similar to those in the 30 varieties, where the average Ho (observed heterozygosity) was 71.48%, and the average DP (discriminative power) was 82.77%, preliminarily indicating the stability of the 487 MNPs. On the other hand, clustering results based on the 487 MNPs were also generally consistent with those based on the 26,147 SNPs, as well as those based on phenotypic traits, and initial grouping, likewise, further indicating the robust capability of our MNPs in variety discrimination, which is similar to their correspondence with numerous SNPs. Therefore, our MNP markers have great potential in the accurate and rapid identification of chrysanthemum varieties, and, accordingly, in fostering breeding innovation and promoting chrysanthemum marketing. Full article
(This article belongs to the Special Issue New Advances in Molecular Biology of Horticultural Plants)
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Figure 1

Figure 1
<p>Cluster and correlation analysis of 30 varieties based on 36 phenotypic characteristics: (<b>a</b>) correlation analysis; (<b>b</b>) cluster correlation analysis. Note: C-1~C-30: variety code (<a href="#horticulturae-10-00845-t001" class="html-table">Table 1</a>); *, **: significant correlation with <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.01, respectively.</p>
Full article ">Figure 2
<p>Genetic analysis of 30 varieties based on 31,448 SNPs concluded from simplified genome sequencing data: (<b>a</b>) phylogenetic tree; (<b>b</b>) principal component analysis; (<b>c</b>) line chart of error rate of cross-validation; (<b>d</b>) population structure diagram (K = 2). Note: C-1~C-30: variety code; G1–G5: group code (<a href="#horticulturae-10-00845-t001" class="html-table">Table 1</a>).</p>
Full article ">Figure 3
<p>Features of the 487 MNP(multiple nucleotide polymorphism) markers of chrysanthemum. (<b>a</b>) Statistical analysis of 487 MNP marker lengths after primer amplification; (<b>b</b>) Statistics of the number of MNP markers corresponding to the number of high-frequency SNPs contained in each of the 487 MNP markers; (<b>c</b>) Statistical analysis of the distribution of 487 MNP markers on chromosomes; (<b>d</b>) Compare the genetic distance of 30 varieties pairwise based on 487 MNP markers and perform logarithmic statistics; (<b>e</b>) Evaluate the discriminative ability of each MNP (MNP001-MNP487 represent the numbering of 487 MNP markers respectively).</p>
Full article ">Figure 4
<p>Genetic relationship analysis of 30 chrysanthemum varieties. (<b>a</b>) Heat map of genetic similarity between any two of the thirty varieties. (<b>b</b>) Phylogenetic tree of chrysanthemum based on 487 MNP sequences from 30 varieties. Note: C-1~C-30: variety code (<a href="#horticulturae-10-00845-t001" class="html-table">Table 1</a>).</p>
Full article ">Figure 5
<p>Phylogenetic tree of 136 chrysanthemum varieties based on 487 MNP sequences. Note: C-31~C-166: variety code (<a href="#app1-horticulturae-10-00845" class="html-app">Table S1</a>). The same color represents the gathering of varieties together.</p>
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13 pages, 3428 KiB  
Article
Development of Multiple Nucleotide Polymorphism Molecular Markers for Enoki Mushroom (Flammulina filiformis) Cultivars Identification
by Fei Liu, Shi-Hui Wang, Ding-Hong Jia, Hao Tan, Bo Wang and Rui-Lin Zhao
J. Fungi 2023, 9(3), 330; https://doi.org/10.3390/jof9030330 - 7 Mar 2023
Cited by 11 | Viewed by 2547
Abstract
The enoki mushroom (Flammulina filiformis) is one of the most important and popular edible mushrooms commercially in China. However, traditional mushroom cultivar identification is challenging due to poor accuracy, heavy workloads, and low reproducibility. To overcome this challenge, we developed a [...] Read more.
The enoki mushroom (Flammulina filiformis) is one of the most important and popular edible mushrooms commercially in China. However, traditional mushroom cultivar identification is challenging due to poor accuracy, heavy workloads, and low reproducibility. To overcome this challenge, we developed a method for identifying F. filiformis strains using multiple nucleotide polymorphism sequencing (MNP-seq). This involved screening 179 universal MNP markers based on whole-genome sequencing data, constructing an MNP sequence library, and performing multiplex PCR amplification and high-sequencing. We further screened 69 core MNP markers and used them to build a neighbor-joining (NJ) phylogenetic tree of 232 cultivated and wild strains. Our analysis showed that all cultivars could be accurately separated by computing genetic similarity values and that the cultivars could be separated into 22 distinct evolutionary pedigrees. The specific value of genetic similarity can be used as the standard to distinguish F. filiformis cultivars, however, it needs to be comprehensively defined by the additional phenotype and biological characteristics of those strains in the future work. Full article
(This article belongs to the Special Issue Edible and Medicinal Macrofungi)
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Figure 1

Figure 1
<p>Distribution of the number of detected MNP markers in <span class="html-italic">Flammulina filiformis</span> strains.</p>
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<p>Phylogenetic tree of <span class="html-italic">F. filiformis</span> based on 69 core MNP sequences from 232 <span class="html-italic">F. filiformis</span> strains. For each strain, the innermost color ring represents the pedigree of the strain, the second ring indicates pileus color, and the outer ring indicates the original source of the strain.</p>
Full article ">Figure 3
<p>The heatmap of the <span class="html-italic">GS</span> values of pairwise comparison between 22 pedigrees and all cultivars.</p>
Full article ">
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