Literature DB >> 30131900

Isolation and characterization of microsatellite loci for Prunus mongolica (Rosaceae).

Yu-Chen Cheng1, De-Jian Zhang2, Zhan-Yuan Lu1, Xue-Song Ye1, Jian-Guo Wang1, Ping Sun3, Bao-Wei Zhang3.   

Abstract

PREMISE OF THE STUDY: Microsatellite primers were developed in Prunus mongolica (Rosaceae), a relict flora endemic in arid areas of the Asian interior, to investigate the genetic diversity, phylogeography, population structure, and history of the species. METHODS AND
RESULTS: Fifty-one microsatellite loci, including di-, tri-, and tetranucelotide repeats, were identified using transcriptome sequencing and bioinformatic screening. The number of alleles ranged from seven to 11 and the levels of observed and expected heterozygosity ranged from 0.545 to 1.000 and 0.600 to 0.989, respectively. Most of the primers also amplified in a group of congeneric species (P. triloba, P. davidiana, P. persica, P. cerasifera, and P. serrulata).
CONCLUSIONS: This set of microsatellite loci is useful for studying the genetic diversity of P. mongolica. In addition, they can also be used to investigate the population structure, phylogeography, and landscape genetic patterns of congeneric species.

Entities:  

Keywords:  Prunus mongolica; Rosaceae; genetic diversity; microsatellite; polymorphism

Year:  2018        PMID： 30131900      PMCID： PMC6025809          DOI： 10.1002/aps3.1158

Source DB:  PubMed          Journal:  Appl Plant Sci        ISSN： 2168-0450            Impact factor:   1.936


Prunus mongolica Maxim. (Rosaceae) is a relict flora that is endemic to arid areas of the Asian interior. Prunus mongolica was listed in the China Plant Red Data Book as endangered; this has resulted from mining, firewood collection, industrialization, and urbanization, which have led to habitat loss and fragmentation (Fu, 1992). Scientific conservation requires a full understanding of genetic diversity, phylogeny, and population structure. Genetic studies are limited in P. mongolica; therefore, it is necessary to develop microsatellite loci for use in the species. Microsatellites, also known as simple sequence repeats (SSRs), have been widely used as DNA markers in studies of population genetics, phylogeography, landscape genetics, and parentage analysis due to their high level of polymorphism, putative neutrality, codominant inheritance, and ease of scoring (Moriguchi et al., 2015). In this study, we isolated and characterized 51 polymorphic microsatellite loci for P. mongolica by transcriptome sequencing and bioinformatic screening.

METHODS AND RESULTS

Total RNA was extracted from fresh leaves of one individual of P. mongolica using the EasyPure Plant RNA Kit (TransGen Biotech Inc., Beijing, China) following the manufacturer's protocol. A cDNA library was prepared using the TruSeq Stranded Total RNA Sample Prep Kit (Illumina, San Diego, California, USA) and sequenced on a HiSeq 3000 sequencing platform (Illumina). Extra (adapters and other Illumina‐specific sequences) and low‐quality sequences were removed from the raw data using the software Trim Galore version 0.4.4 (Li et al., 2015). FastQC version 0.11.5 software (Yang et al., 2013) was used to analyze the quality control of the pre‐processed data. A total of 53,866 contigs were obtained by de novo assembly using Trinity version 2.3.2 (Grabherr et al., 2011). SSR detection was performed with MISA version 1.0 (Thiel et al., 2003) with the following criteria: ≥6 repeat units for dinucleotides, ≥5 for tri‐, and ≥4 for tetra‐, penta‐, and hexanucleotides. Primer sets were designed with Primer3 using the default parameters (Koressaar and Remm, 2007; Untergasser et al., 2012). A total of 19,498 SSRs were identified from 15,837 contigs, and 6069 primer sets were designed. Two hundred primer sets that amplified di‐, tri‐, and tetranucelotide repeats with ≥6, ≥5, and ≥4 repeat units, respectively, were selected to test for polymorphism. Raw transcriptome data were deposited in the National Center for Biotechnology Information Short Read Archive (BioProject no. PRJNA418876, BioSample no. SAMN08038438). Because of habitat loss, only 32 P. mongolica individuals were sampled from three populations (Appendix 1). Total genomic DNA was extracted from 0.15 mg of leaf tissue using a modified cetyltrimethylammonium bromide (CTAB) method (Wang et al., 2010). All PCRs were performed in total volumes of 15 μL: 1 μL of DNA, 5.5 μL of sterilized deionized water, 7.5 μL of 2× EasyTaq PCR Supermix (TransGen Biotech Inc.), and 0.5 μL each of forward and reverse primers (each forward primer was fluorescently labeled with FAM, HEX, or TAMRA). PCR amplification was carried out using the following thermal conditions: initial denaturation for 5 min at 94°C; then 35 cycles of denaturation for 30 s at 94°C, annealing for 30 s at the annealing temperature (Table 1), and 20 s at 72°C; followed by a final extension for 10 min at 72°C. PCR products were genotyped on an ABI PRISM 3730 genetic analyzer (Applied Biosystems, Waltham, Massachusetts, USA) with a GeneScan 500 size standard (Applied Biosystems). Allele peaks were scored using GeneMarker version 1.3 (SoftGenetics, State College, Pennsylvania, USA). MICRO‐CHECKER version 2.2.3 (van Oosterhout et al., 2004) was used to detect genotyping errors due to null alleles, stuttering, or allele dropout. The number of effective alleles and levels of observed and expected heterozygosity were calculated using GENETIX version 4.0 (Belkhir et al., 2001). Deviation from Hardy–Weinberg equilibrium and linkage disequilibrium was tested with GENEPOP version 3.4 (Rousset, 2008).
Table 1

Characteristics of 51 microsatellite loci developed in Prunus mongolica

LocusPrimer sequences (5′–3′)Repeat motifAllele size range (bp) T a (°C)FunctionGenBank accession no.
TR3320F: CGCCTCCTCTCTCTCTCTA(TC)19 127–17555EIN3‐binding F‐box protein 1 MG682092
R: CCCAGAAAAGATTAAGGCTAC
TR5622F: AAACGAGAGCAGGATCAATA(CAA)8 127–17955B3 domain‐containing transcription factor MG682093
R: CGTAGTTTGGGTTGGAGTC
TR9296F: GCAGAGAACTTATCGCTTATG(TTTA)6 133–20155Glyoxylate/hydroxypyruvate reductase HPR3 MG682094
R: CATCTGGAAAGTGAATCTGAG
TR10339F: ACTCTTCTCTCCATTGAGTCC(CT)20 105–14155Protein MIZU‐KUSSEI 1 MG682095
R: GTCCTCATCTGGGAAATTTAG
TR10936F: AGAAGATAGAGGGATGGGAGT(AG)21 153–18156GATA transcription factor 11 MG682096
R: CTCTCTGCAACAACCAAAAC
TR12256F: GAGAGAGAAAAATTGCAGTCC(TCT)7 131–17755Unknown MG682097
R: CCATGGAGATTAAGAGGAAGA
TR12298F: CAATAGCCAAAACCCTCTC(TC)6 123–16954DNA‐directed RNA polymerases II, IV, and V MG682098
R: GCCTTCCATTTCTCTATGTCT
TR12728F: AAATCTAAGCCCACGTACATC(GA)18 153–19356Protein‐tyrosine‐phosphatase MKP1 MG682099
R: CACCCTGGTAACTCATAATCA
TR14394F: TGAGCTACAGACAGGTGAACT(GAA)7 143–16955Transcription factor bHLH25 MG682100
R: ACACAACACACCACTTTTCTC
TR16484F: GTGTTGGCTAGAGTGACTTTG(TC)8 157–19556UDP‐glycosyltransferase 74F2 MG682101
R: CGCCACACCTAAGTTTTT
TR17663F: TCATCTCCTCTACCTTTTTCC(CT)19 115–16955Unknown MG682102
R: GGTCATGATCTCTGCTTGATA
TR17745F: CCTTCCATTTCTCTGTTCTCT(TC)21 111–14955Aquaporin PIP1‐2‐like MG682103
R: CACTCAACCAAATCATAGCTC
TR18640F: AACTAGAGAGAGAGACGCACA(GA)16 91–13955Succinate‐CoA ligase [ADP‐forming] subunit beta, mitochondrial MG682104
R: GTTCTTTCGCATTCTCAGAC
TR18685F: CAGAAGTCAGAGCAGAGTCAG(ACAG)6 139–16955VQ motif‐containing protein 9 MG682105
R: CTACCGGGAGATATTTCAGAT
TR18807F: GTCCTTAAGCAGCCCAGTA(CT)20 121–17355Probable acyl‐activating enzyme 17, peroxisomal MG682106
R: AGAGCTTCGATGTCAGAGAC
TR19174F: CTTGAGAGGAGGAAAAAGAAG(AGA)8 145–21755Transcription factor PRE6‐like MG682107
R: GCTGTCTGGGGTTATAAATTC
TR19201F: TATTGCACTCTCCTATGAACC(CAG)7 139–16755Auxin response factor 7 (LOC110749255), transcript variant X2 MG682108
R: GTTGAGACTCTTGCTGTTGTT
TR20374F: CTTCCATCCACACTTACAT(GA)8 149–20755Uncharacterized protein At1g04910 MG682109
R: GAGGGACTGAATAAACCTGAG
TR21328F: AAGGTGAAGGGTGAAGATACT(GA)12 141–19555Magnesium‐chelatase subunit ChlH, chloroplastic MG682110
R: AAACACAATCACAGAGAGAGC
TR21918F: CCTCCTTCTGTCTCTCTCTTC(CT)12 125–18755E3 ubiquitin‐protein ligase RNF144A MG682111
R: CAGTTAAACCACCAGCACTAC
TR20030F: ATAACCCCAAGCTCTCTCTC(CTC)8 131–16155 Prunus persica receptor‐like protein 51 MG682112
R: AGGAGGTAGGAGATGGAGTG
TR22071F: CAGAAGAAAAAGAAGGAGAGC(GAA)7 131–16155 Prunus avium cysteine proteinase inhibitor 12‐like, transcript variant X6 MG682113
R: TGTCAACCATACTCACCTCAT
TR23600F: CAGCTCTCTCTTTCTCTCTCC(TC)8 141–18355Probable serine/threonine‐protein kinase DDB_G0282963 MG682114
R: ACTTCCACTACCAGAATCCAT
TR24122F: GGTCTGTTAATGGAGGTTCTT(CT)13 133–16956UDP‐N‐acetylglucosamine–dolichyl‐phosphate MG682115
R: CACCAATATTGAGAGAGAGAC
TR25043F: AACCAACCCAACCCACAC(ACAT)6 111–16557 N‐acetylglucosaminephosphotransferase MG682116
R: GGGGGAGAACCTATCTTATATT
TR25166F: CAGGCTCTGTAGTTCGATTC(TCT)8 147–17955Gibberellin 20 oxidase 1 MG682117
R: TGTATGTTAGTAAGGCGAAGG
TR25414F: CAGCAGAGAGACCAGAATAGA(TCCA)6 103–12555Uncharacterized protein At2g17340‐like MG682118
R: GAGCATTATAGCTGTGTGTGG
TR26908F: TATCTTCATCGCTCTCTTCTG(CAT)10 111–14955Galactomannan galactosyltransferase 1 MG682119
R: CTGGGGTCTTAATTAGACGTT
TR27527F: CACCAAACCTTTCTCTCTCTT(CT)27 117–16554Transcription factor HY5‐like, transcript variant X1 MG682120
R: ACAAAGTCGATCAGCATCTC
TR29327F: CTGCCGTCTATCTCTCTCTCT(CT)6 113–14556Probable fructokinase‐4 MG682121
R: CAAGTTTGGTGCTCGTATCTA
TR30962F: CTATGGGTCTGGTTATGGAA(GGA)11 101–16155Cytosolic 5′‐nucleotidase 3, transcript variant X5 MG682122
R: CTCCCAGCCAATATCTCAG
TR31156F: CTCTGCCTCTTTCTCTTCTCT(AG)14 117–15555Putative glycine‐rich cell wall MG682123
R: ATCCCCCTACTTTCTTGC
TR31399F: GAAGAACAAGAAGGGCAAG(AG)20 117–16755Structural protein 1 MG682124
R: CAACACTAACACCACCAGATT
TR31622F: AAGCCAGAACCCTTATCTCTA(AAC)5 125–17955Unknown MG682125
R: GCCTGAATTGAGTGAACTGTA
TR32795F: CAAGTTGATTTCTTCCTCTCC(AGA)13 119–15555Peamaclein‐like MG682126
R: TATAACGCCTCTCTCTGTTGA
TR33107F: AAAACAGAGAGGACGATGG(TTGT)6 139–17155BEL1‐like homeodomain protein 9, transcript variant X1 MG682127
R: GCTCTAGGTGTTGGGTTAAAT
TR35128F: GCCTGAATTGAGTGAACTGTA(TTG)8 129–18156Glutamine–fructose‐6‐phosphateaminotransferase [isomerizing] 2 MG682128
R: AAGCCAGAACCCTTATCTCTA
TR35536F: CTCTTGTGTTCATCGGCTAC(TGA)6 153–17555Calcium‐dependent protein kinase 13 MG682129
R: ATCTCACCCGTCTCTTTATTC
TR37605F: CTTTCTAACCCCTCTGTGTCT(TC)21 117–15355BEL1‐like homeodomain protein 9, transcript variant X2 MG682130
R: TATGAACAGCTTACCTTCGTC
TR37669F: CCTCTTTGATCTCCTTCCATA(TCT)8 141–18155Glutamate synthase [NADH], amyloplastic, transcript variant X3 MG682131
R: GAAGACGACCTTTTTCATACC
TR37811F: GATCAAGGAGGAAGACAAAGT(AG)26 105–14555Heterogeneous nuclear ribonucleoprotein 1 MG682132
R: GCAGAGAGAGAGAAAGAGGAA
TR38169F: GATCATGAAGTTGCAGAAGAG(TGG)10 143–18556Probable WRKY transcription factor 12 MG682133
R: ATGTGCCCTTCCTGTGAG
TR38361F: GGATGTGCTTTACTGGAGAA(TGA)8 127–15556Probable phospholipid‐transporting ATPase MG682134
R: CTCCTTCTAATCCCTTCAGC
TR41171F: TAGAAACTATGTGCGTGTGTG(GA)15 123–175554, transcript variant X2 MG682135
R: CTTCAGGGAGGCTATAAATTC
TR42254F: ATTGCTTGCTCTGTCTCTGTA(AAG)5 141–18155Homeobox‐leucine zipper protein HOX11‐like, transcript variant X2 MG682136
R: CTTATTACCCACTACCCCAAC
TR42785F: TCTCTGAAATTCTCTCTGCTC(GTGC)6 117–14755Formin‐like protein 4 MG682137
R: TCTTTCTTCCTCCTCTCTCAC
TR46482F: CCACCTTCTTATAGCGACTG(TATG)6 133–19355Probable E3 ubiquitin‐protein ligase RZFP34, transcript variant X4 MG682138
R: AAGTACGATACAAGCACTCCA
TR47534F: CGATCTCCTTCCAGACCT(GTG)6 119–15755Auxin efflux carrier component 1 MG682139
R: CGACATCCTCAAGAACATCTA
TR47552F: ACAATCTCTTCGTCTCTGGTT(AG)19 121–17754UDP‐glycosyltransferase 76E2‐like, transcript variant X2 MG682140
R: GCTCCAGTGTACGTTGTAAG
TR50321F: AAGACCACTCACAACTCCAC(CTT)8 263–29355Cyclic nucleotide‐gated ion channel 1‐like, transcript variant X1 MG682141
R: CTTTGAAACACTGTTCTGGTC
TR52495F: TGATGAGCTTGCCATTCT(GAA)8 147–17355G‐box‐binding factor 4, mRNA MG682142
R: CTTCTTGTCCGCTAATTCAC

T a = annealing temperature.

Characteristics of 51 microsatellite loci developed in Prunus mongolica T a = annealing temperature. A total of 51 primer sets produced consistent amplifications in all samples and were polymorphic (Table 1). These 51 loci had high numbers of effective alleles and high levels of observed and expected heterozygosity, ranging from seven to 11, 0.545 to 1.000, and 0.600 to 0.989, respectively (Table 2). Cross‐amplification of these 51 primer sets was tested for in P. triloba Lindl., P. davidiana (Carrière) Franch., P. persica (L.) Batsch, P. cerasifera Ehrh., and P. serrulata Lindl.; most primers amplified in these five species except P. triloba (P. triloba: 35/51; P. davidiana: 43/51; P. persica: 46/51; P. cerasifera: 45/51; P. serrulata: 41/51; Table 3).
Table 2

Genetic diversity of 51 microsatellite loci in three populations of Prunus mongolica.a

LocusWulanchabu population (n = 10)Chifeng population (n = 10)Eerduosi population (n = 12)
A e H o H e A e H o H e A e H o H e
TR33208.5000.8890.9228.5000.8890.8699.0001.0000.925
TR562211.0001.0000.92810.5001.0000.9639.5000.8330.880
TR92967.0001.0000.9678.0001.0000.9707.0001.0000.925
TR1033911.0001.0000.90011.0001.0000.83210.0001.0000.935
TR1093610.5001.0000.92210.0001.0000.8899.5000.9090.926
TR1225610.0000.8890.8379.5000.8000.6009.5000.8000.847
TR1229811.0001.0000.94110.5001.0000.8589.5001.0000.906
TR1272810.0001.0000.94810.0001.0000.9549.0001.0000.874
TR1439411.0000.7780.89510.5000.9000.7589.5000.7500.699
TR1648410.5001.0000.90810.0001.0000.9219.5001.0000.909
TR1766310.5001.0000.98010.5001.0000.9619.0001.0000.960
TR1774511.0001.0000.97410.5001.0000.9539.5001.0000.928
TR1864010.5001.0000.98910.5001.0000.95310.0001.0000.939
TR1868511.0000.8750.90810.0000.8000.8589.0000.8330.790
TR188079.5001.0000.94210.5001.0000.9679.0001.0000.939
TR1917411.0000.8890.81010.5001.0000.8539.5001.0000.938
TR1920111.0001.0000.85811.0000.9000.88410.0000.9170.888
TR2003011.0001.0000.94110.5001.0000.9539.5000.9170.928
TR2037410.5001.0000.97410.0001.0000.9639.5001.0000.952
TR2132811.0001.0000.96311.0001.0000.95810.0001.0000.960
TR2191811.0001.0000.95410.5001.0000.9219.5001.0000.960
TR2207111.0000.8890.81710.5000.9000.9429.5000.7500.928
TR2360010.5001.0000.90810.5001.0000.9359.0001.0000.949
TR241229.0001.0000.9228.5000.9000.9379.5001.0000.925
TR2504310.5000.7780.88210.0000.6000.7059.5000.7270.870
TR2516611.0001.0000.91510.5001.0000.9639.5001.0000.920
TR2541410.5001.0000.7339.5001.0000.8329.0000.8890.771
TR2690810.0000.9000.88410.5000.8890.7919.5000.9090.905
TR2752711.0000.9000.86311.0000.9000.83710.0000.8330.920
TR2932710.0001.0000.92810.0001.0000.8769.0001.0000.952
TR309629.5001.0000.8679.0001.0000.9358.5001.0000.932
TR3115610.0000.8890.97410.0001.0000.9159.0001.0000.931
TR3139910.5001.0000.96110.0001.0000.9799.5001.0000.935
TR3162211.0000.8000.88411.0001.0000.91610.0000.8330.851
TR3279511.0000.9000.92111.0001.0000.92610.0000.9170.891
TR3310711.0000.9000.91111.0000.9000.87410.0000.8330.909
TR3512811.0000.7780.81710.5001.0000.9009.5000.7500.855
TR3553610.5000.7780.88210.0000.9000.8379.5000.9090.900
TR3760511.0000.9000.92611.0001.0000.89510.0001.0000.935
TR3766910.5000.8890.86910.5000.8890.8899.0001.0000.899
TR3781110.5001.0000.93510.5001.0000.9419.0001.0000.957
TR3816910.5001.0000.94110.5001.0000.9619.0000.9170.935
TR3836110.0001.0000.85810.5001.0000.7789.5001.0000.840
TR4117110.5001.0000.96710.0001.0000.9808.5001.0000.931
TR4225411.0000.9000.92111.0000.8000.92110.0000.5830.851
TR4278510.0000.7500.83310.0000.8750.8678.0000.8330.866
TR464829.5001.0000.94110.5001.0000.9238.0000.8330.848
TR475349.0001.0000.8178.5000.7780.6808.5000.5560.804
TR4755210.0001.0000.96710.0001.0000.9419.0001.0000.948
TR503219.0000.8000.87410.5000.8570.8688.5000.5450.827
TR524959.5000.5560.8439.5001.0000.9029.0000.8000.805

A e = number of effective alleles; H e = expected heterozygosity; H o = observed heterozygosity.

Locality and voucher information are provided in Appendix 1.

Table 3

Cross‐amplification results for the 51 microsatellite loci developed in Prunus mongolica in five Prunus species.a , b

Locus P. triloba P. davidiana P. persica P. cerasifera P. serrulata
TR10339—163–195115–147133–171117–151
TR10936—137–149119–146125–167149–161
TR12298—103–115—132–144105–121
TR12728—110–122105–117105–127120–132
TR16484—153–165143–155133–145113–125
TR17663—131–140130–142—120–129
TR17745120–126110–122123–135133–145125–137
TR18640135–147153–159133–145131–143—
TR18807140–149143–155133–145113–119131–143
TR20374115–127151–163157–169153–165151–159
TR21328111–131115–136113–125117–129—
TR21918101–121110–130115–135118–138111–123
TR23600——80–10091–101—
TR24122——130–150139–159133–153
TR3320———121–133—
TR27527140–149145–160132–144141–153135–147
TR29327133–145135–147130–142132–138—
TR31156————141–161
TR31399131–151111–131143–163141–161—
TR37605125–145126–146121–141122–142123–143
TR37811131–161121–151111–141146–176132–162
TR41171131–191112–142123–153119–149145–175
TR47552121–141122–142111–131123–143101–131
TR26908113–129133–149113–125115–127131–143
TR30962111–127131–143127–139123–139135–147
TR32795125–137123–126147–159131–161126–138
TR19174103–123121–133116–128113–129132–144
TR19201142–154131–161125–137126–138141–171
TR20030115–135117–139123–135134–146137–149
TR25166113–125132–144115–135162–182133–145
TR31622151–163131–143133–163142–154125–137
TR35128117–129146–158135–155132–144172–184
TR35536137–157117–129153–165135–155127–167
TR37669123–138121–133125–137122–134115–127
TR38169——125–137——
TR38361133–149131–147115–127124–136125–137
TR42254122–134117–129115–127123–135116–128
TR47534123–135115–127132–144124–136125–137
TR52495117–129112–124113–125117–129123–135
TR5622121–151124–136142–154141–153123–135
TR12256—103–115105–117113–133132–152
TR14394143–155124–136121–130123–135141–153
TR22071122–155114–126142–154121–133113–125
TR50321———123–143114–124
TR18685113–125121–133135–147123–135141–153
TR25043—117–137132–144124–136—
TR25414133–145123–139121–133142–158152–164
TR33107121–133143–155117–139—113–128
TR42785—————
TR46482253–265221–236232–244—215–227
TR9296——141–156122–134—

— = unsuccessful amplification.

Locality and voucher information are provided in Appendix 1.

N = 5 for all populations.

Genetic diversity of 51 microsatellite loci in three populations of Prunus mongolica.a A e = number of effective alleles; H e = expected heterozygosity; H o = observed heterozygosity. Locality and voucher information are provided in Appendix 1. Cross‐amplification results for the 51 microsatellite loci developed in Prunus mongolica in five Prunus species.a , b — = unsuccessful amplification. Locality and voucher information are provided in Appendix 1. N = 5 for all populations.

CONCLUSIONS

In our study, a total of 19,498 SSRs were identified from 15,837 contigs, and 6069 microsatellite primers were designed. Among the identified SSR loci, 51 microsatellite primer pairs were polymorphic. The 51 novel microsatellite markers can be used to study the genetic diversity, landscape genetics, and phylogeography of the species. The microsatellite primers can also be used to genotype congeneric species (P. triloba, P. davidiana, P. persica, P. cerasifera, and P. serrulata).
Species n Collection localityGeographic coordinatesVoucher IDa Collector
Prunus mongolica Maxim.10Wulanchabu, Inner Mongolia, China41°56′N, 110°28′E201607‐Pmon‐popI‐AHBZhanyuan Lu
10Chifeng, Inner Mongolia, China43°23′N, 118°40′E201607‐Pmon‐popII‐AHBZhanyuan Lu
12Eerduosi, Inner Mongolia, China39°35′N, 106°46′E201607‐ Pmon‐popIII‐AHBZhanyuan Lu
P. triloba Lindl.5Hefei, Anhui, China31°52′N, 117°11′E201705‐Ptri‐AHBXin Hong
P. davidiana (Carrière) Franch.5Hefei, Anhui, China31°52′N, 117°11′E201705‐Pdav‐AHBXin Hong
P. persica (L.) Batsch5Wuhan, Hubei, China30°32′N, 114°24′E201705‐Pper‐AHBXin Hong
P. cerasifera Ehrh.5Hefei, Anhui, China31°52′N, 117°11′E201705‐Pcer‐AHBXin Hong
P. serrulata Lindl.5Hefei, Anhui, China31°52′N, 117°11′E201705‐Pser‐AHBXin Hong

n = sample size.

Vouchers are deposited at the herbarium of Anhui University (ANU), Hefei, Anhui Province, China.

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7.  PEAT: an intelligent and efficient paired-end sequencing adapter trimming algorithm.

Authors:  Yun-Lung Li; Jui-Cheng Weng; Chiung-Chih Hsiao; Min-Te Chou; Chin-Wen Tseng; Jui-Hung Hung
Journal:  BMC Bioinformatics       Date:  2015-01-21       Impact factor: 3.169

8.  Full-length transcriptome assembly from RNA-Seq data without a reference genome.

Authors:  Manfred G Grabherr; Brian J Haas; Moran Yassour; Joshua Z Levin; Dawn A Thompson; Ido Amit; Xian Adiconis; Lin Fan; Raktima Raychowdhury; Qiandong Zeng; Zehua Chen; Evan Mauceli; Nir Hacohen; Andreas Gnirke; Nicholas Rhind; Federica di Palma; Bruce W Birren; Chad Nusbaum; Kerstin Lindblad-Toh; Nir Friedman; Aviv Regev
Journal:  Nat Biotechnol       Date:  2011-05-15       Impact factor: 54.908
  8 in total

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