High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)

Lady’s slipper orchids (<i>Paphiopedilum</i> spp.) are highly valuable within the flower industry. Recently, both <i>Paphiopedilum concolor</i> and <i>Paphiopedilum hirsutissimum</i> (2n = 2x = 26) have been widely used for hybrid parents, ornamental, and economic...

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Main Authors: Dong-Mei Li, Gen-Fa Zhu
Format: Article
Language:English
Published: MDPI AG 2022-09-01
Series:Horticulturae
Subjects:
Online Access:https://www.mdpi.com/2311-7524/8/9/842
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author Dong-Mei Li
Gen-Fa Zhu
author_facet Dong-Mei Li
Gen-Fa Zhu
author_sort Dong-Mei Li
collection DOAJ
description Lady’s slipper orchids (<i>Paphiopedilum</i> spp.) are highly valuable within the flower industry. Recently, both <i>Paphiopedilum concolor</i> and <i>Paphiopedilum hirsutissimum</i> (2n = 2x = 26) have been widely used for hybrid parents, ornamental, and economic purposes. However, high-density genetic maps and leaf traits related to quantitative trait loci (QTLs) in these two <i>Paphiopedilum</i> species have been poorly studied. Herein, an interspecific F1 population of 95 individuals was developed from the cross between <i>P. concolor</i> and <i>P. hirsutissimum</i> with contrasting leaf length (LL), leaf width (LW), leaf thickness (LT), and leaf number (LN). RNA extracted from the F1 population and their parents was subjected to high-throughput RNA sequencing. Approximately 745.59 Gb of clean data were generated, and were assembled into 349,730 transcripts and 185,089 unigenes. In total, 165,196 high-resolution polymorphic single nucleotide polymorphism (SNP) markers were initially identified. Finally, 8410 SNP markers satisfied the requirements and were used to construct a genetic map. The integrated map contained 13 linkage groups (LGs) and spanned 1616.18 cM, with an average distance of 0.19 cM between adjacent markers. QTL analysis in the F1 population identified 12 QTLs linked to four leaf-related traits, including LL, LW, LT, and LN. These QTLs by composition interval mapping, explained 11.86% to 21.58% of the phenotypic variance, and were distributed on eight LGs, but not on LGs 4, 6, 8, 12, and 13. Furthermore, 25 unigenes were identified via BLAST searches between the SNP markers in the QTL regions and our assembled transcriptome, of which 11 unigenes were enriched with 59 gene ontology (GO) terms. The information generated in this study will be useful for candidate genes for further molecular regulation studies on leaf traits, future marker-assisted selection of leaf ornamental improvement breeding, genome assembly, and comparative genome analyses.
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spelling doaj.art-dfb7e44249af4435ab8b26d53c8cfb122023-11-23T16:34:32ZengMDPI AGHorticulturae2311-75242022-09-018984210.3390/horticulturae8090842High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)Dong-Mei Li0Gen-Fa Zhu1Guangdong Key Lab of Ornamental Plant Germplasm Innovation and Utilization, Environmental Horticulture Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, ChinaGuangdong Key Lab of Ornamental Plant Germplasm Innovation and Utilization, Environmental Horticulture Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, ChinaLady’s slipper orchids (<i>Paphiopedilum</i> spp.) are highly valuable within the flower industry. Recently, both <i>Paphiopedilum concolor</i> and <i>Paphiopedilum hirsutissimum</i> (2n = 2x = 26) have been widely used for hybrid parents, ornamental, and economic purposes. However, high-density genetic maps and leaf traits related to quantitative trait loci (QTLs) in these two <i>Paphiopedilum</i> species have been poorly studied. Herein, an interspecific F1 population of 95 individuals was developed from the cross between <i>P. concolor</i> and <i>P. hirsutissimum</i> with contrasting leaf length (LL), leaf width (LW), leaf thickness (LT), and leaf number (LN). RNA extracted from the F1 population and their parents was subjected to high-throughput RNA sequencing. Approximately 745.59 Gb of clean data were generated, and were assembled into 349,730 transcripts and 185,089 unigenes. In total, 165,196 high-resolution polymorphic single nucleotide polymorphism (SNP) markers were initially identified. Finally, 8410 SNP markers satisfied the requirements and were used to construct a genetic map. The integrated map contained 13 linkage groups (LGs) and spanned 1616.18 cM, with an average distance of 0.19 cM between adjacent markers. QTL analysis in the F1 population identified 12 QTLs linked to four leaf-related traits, including LL, LW, LT, and LN. These QTLs by composition interval mapping, explained 11.86% to 21.58% of the phenotypic variance, and were distributed on eight LGs, but not on LGs 4, 6, 8, 12, and 13. Furthermore, 25 unigenes were identified via BLAST searches between the SNP markers in the QTL regions and our assembled transcriptome, of which 11 unigenes were enriched with 59 gene ontology (GO) terms. The information generated in this study will be useful for candidate genes for further molecular regulation studies on leaf traits, future marker-assisted selection of leaf ornamental improvement breeding, genome assembly, and comparative genome analyses.https://www.mdpi.com/2311-7524/8/9/842<i>Paphiopedilum</i>RNA sequencingsingle nucleotide polymorphismgenetic mapquantitative trait locileaf traits
spellingShingle Dong-Mei Li
Gen-Fa Zhu
High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)
Horticulturae
<i>Paphiopedilum</i>
RNA sequencing
single nucleotide polymorphism
genetic map
quantitative trait loci
leaf traits
title High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)
title_full High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)
title_fullStr High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)
title_full_unstemmed High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)
title_short High-Density Genetic Linkage Map Construction and QTLs Identification Associated with Four Leaf-Related Traits in Lady’s Slipper Orchids (<i>Paphiopedilum concolor</i> × <i>Paphiopedilum hirsutissimum</i>)
title_sort high density genetic linkage map construction and qtls identification associated with four leaf related traits in lady s slipper orchids i paphiopedilum concolor i i paphiopedilum hirsutissimum i
topic <i>Paphiopedilum</i>
RNA sequencing
single nucleotide polymorphism
genetic map
quantitative trait loci
leaf traits
url https://www.mdpi.com/2311-7524/8/9/842
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