Development of 29 microsatellite markers for <i>Osmanthus fragrans</i> (Oleaceae), a traditional fragrant flowering tree of China

Zhang, Zhirong; Fan, Dashan; Guo, Shi-Quan; D, Li; Zhang, Zhiyong

doi:10.3732/ajb.1100241

Cited by 8 publications

(7 citation statements)

References 9 publications

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“…A total of 100 SSR primers from the developed primers based on high-throughput transcriptome sequencing data [8,27] were randomly selected and 15 primers with stable polymorphic bands were screened by the DNA mixed pooling method [28]. Then, the 15 specific primers with three universal primers (OSM005, OSM010, OSM038) [29] were used to estimate the genetic diversity and structure of the natural O. serrulatus populations (Table S1). The SSR-PCR amplification was performed on the Applied Biosystems TM Automated Thermal Cycler (Thermo Scientific, Wilmington, DE, USA) as follows: a pre-denaturation at 94 °C for 5 min; followed by 30 cycles of 4 s denaturation at 94 °C, 30 s annealing at 55 °C and 30 s extension at 72 °C; a final extension at 72 °C for 5 min; and then preservation at 4 °C [28].…”

Section: Primer Screening and Pcr Amplificationmentioning

confidence: 99%

Genetic Diversity and Population Structure Revealed by SSR Markers on Endemic Species Osmanthusserrulatus Rehder from Southwestern Sichuan Basin, China

Chen

Pan

Qian

et al. 2021

Forests

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Osmanthus serrulatus Rehder (Oleaceae) is an endemic spring-flowering species in China. It is narrowly distributed in the southwestern Sichuan Basin, and is facing the unprecedented threat of extinction due to problems associated with natural regeneration, habitat fragmentation and persistent and serious human interference. Here, the genetic diversity and population structure of 262 individuals from ten natural populations were analyzed using 18 microsatellites (SSR) markers. In total, 465 alleles were detected across 262 individuals, with a high polymorphic information content (PIC = 0.893). A high level of genetic diversity was inferred from the genetic diversity parameters (He = 0.694, I = 1.492 and PPL = 98.33%). AMOVA showed that a 21.55% genetic variation existed among populations and the mean pairwise Fst (0.215) indicated moderate genetic population differentiation. The ten populations were basically divided into three groups, including two obviously independent groups. Our results indicate that multiple factors were responsible for the complicated genetic relationship and endangered status of O. serrulatus. The concentrated distribution seems to be the key factor causing endangerment, and poor regeneration, human-induced habitat loss and fragmentation seem to be the primary factors in the population decline and further genetic diversity loss. These findings will assist in future conservation management and the scientific breeding of O. serrulatus.

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Section: Primer Screening and Pcr Amplificationmentioning

confidence: 99%

Genetic Diversity and Population Structure Revealed by SSR Markers on Endemic Species Osmanthusserrulatus Rehder from Southwestern Sichuan Basin, China

Chen

Pan

Qian

et al. 2021

Forests

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“…Nine pairs of primers (OSM005、OSM010、OSM011、OSM014、OSM017、OSM023、OSM025、OSM036、0SM052) with high stability and polymorphism were obtained from synthesizing 15 pairs of O . delavayi fluorescence primers [31]. Amplification products were produced by polymerase chain reaction using a PCR machine (Coyote Bio CF-F9677, America), and were submitted to agarose gel electrophoresis (HDL apparatus DL-EP300) to detect the amplification effect.…”

Section: Methodsmentioning

confidence: 99%

Functional androdioecy in the ornamental shrub Osmanthus delavayi (Oleaceae)

Duan

Zheng

et al. 2019

PLoS ONE

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Androdioecy is one of the rarest sexual systems among plants, characterized by males co-occurring with hermaphrodites. Osmanthus delavayi (Oleaceae), an ornamental shrub from southern China, is known to have both male and hermaphrodite individuals, but little is known regarding the breeding system of this species and whether it is functionally androdioecious, and how this potentially evolved. In this study, we explore the characteristics of the breeding system of O . delavayi through the study of phenology, sex ratio, floral organ morphology, pollen number, stigma receptivity, artificial pollination, pollinators, and gene flow within and between populations, while also discussing the evolution and maintenance of androdioecy within the genus. The proportion of males was less than 0.5 and the out-crossing index (OCI) was 5. Morphological androdioecy was observed, with hermaphrodite flowers having fertile pistils, while male flowers had degenerated pistils. Males and hermaphrodites both had large amounts of small and fertile pollen grains, although the pollen number of males was ca. 1.21 × more than that of hermaphrodites, and pollen was generally smaller. Self-pollination was found to produce a much lower fruit set than outcrossing under natural conditions. Gene flow between males and hermaphrodites within a population was greater (1.007) than that between populations (0.753). All these results indicate that O . delavayi is functionally androdioecious, which may be an intermediate state in the evolutionary transition from hermaphroditism to dioecy.

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“…Initial progress in advancing a molecular toolkit for Osmanthus includes the production of microsatellites in O. fragrans through enriched library development and transcriptome sequencing and Osmanthus phylogenies and genetic relationships inferred from chloroplast and nuclear markers. [4][5][6][7][8][9] These studies, which collectively include 22 Osmanthus species, report genomic DNA isolation by the CTAB method of Doyle and Doyle. 10 None of the studies mention amount or purity of isolated DNA; cost, time, or expense of the isolation procedure; or relative ease or difficulty of the species examined.…”

Section: Introductionmentioning

confidence: 99%

Rapid, Effective DNA Isolation from Osmanthus via Modified Alkaline Lysis

Alexander

2016

J Biomol Tech

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Variability of leaf structure and presence of secondary metabolites in mature leaf tissue present a challenge for reliable DNA extraction from Osmanthus species and cultivars. The objective of this study was to develop a universal rapid, effective, and cost-efficient method of DNA isolation for Osmanthus mature leaf tissue. Four different methods were used to isolate DNA from 8 cultivars of Osmanthus. Absorbance spectra, DNA concentration, appearance on agarose gel, and performance in PCR were used to analyze quality, quantity, and integrity of isolated DNA. Methods were ranked in order, based on total quantity, quality, and performance points as the following: 1) solid-phase extraction (SPE), 2) modified alkaline lysis (SDS), 3) cetyltrimethylammonium bromide (CTAB) with chloroform (CHL), and 4) CTAB with phenol/chloroform (PHE). Total DNA, isolated via SPE, showed the least contamination but the lowest mean quantity (9.6 ± 3.4 μg) and highest cost. The highest quantity of DNA was isolated via SDS (117 ± 54.1 μg). SPE and SDS resolved the most individuals on agarose gel, whereas the 2 CTAB methods had poorly resolved gels. All methods except PHE performed well in PCR. Additions to the modified alkaline lysis method increased A260:A230 by up to 59% without affecting yield. With the use of SDS, an average of 1000 μg/g DNA was isolated from fresh leaf tissue of 18 samples in ∼1.5 h at a cost of 0.74 U.S. dollars (USD)/sample. We recommend improved alkaline lysis as a rapid, effective, and cost-efficient method of isolating DNA from Osmanthus species.

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Development of 29 microsatellite markers for Osmanthus fragrans (Oleaceae), a traditional fragrant flowering tree of China

Abstract: These markers will facilitate further studies on the population genetics of O. fragrans and the classification and identification of O. fragrans cultivars.

Cited by 8 publications

References 9 publications

Genetic Diversity and Population Structure Revealed by SSR Markers on Endemic Species Osmanthusserrulatus Rehder from Southwestern Sichuan Basin, China

Genetic Diversity and Population Structure Revealed by SSR Markers on Endemic Species Osmanthusserrulatus Rehder from Southwestern Sichuan Basin, China

Functional androdioecy in the ornamental shrub Osmanthus delavayi (Oleaceae)

Rapid, Effective DNA Isolation from Osmanthus via Modified Alkaline Lysis

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