Use of EST-SSR Markers for Evaluating Genetic Diversity and Fingerprinting Celery (Apium graveolens L.) Cultivars
Abstract
:1. Introduction
2. Results and Discussion
Code | Variety | Type | Species |
---|---|---|---|
C1 | Xuebaiqincai | Local celery | Apium graveolens L. var. dulce |
C3 | Jinhuangqincai | Local celery | Apium graveolens L. var. dulce |
C5 | Jincuifuqin | Local celery | Apium graveolens L. var. dulce |
C6 | Huangxinqin | Local celery | Apium graveolens L. var. dulce |
C8 | Shanghaichunqin | Local celery | Apium graveolens L. var. dulce |
C13 | Tieganqing | Local celery | Apium graveolens L. var. dulce |
C29 | Wuqiangshiganqincai | Local celery | Apium graveolens L. var. dulce |
C37 | Shixinqin | Local celery | Apium graveolens L. var. dulce |
C53 | Duolunshiganqincai | Local celery | Apium graveolens L. var. dulce |
C111 | Lino | Celery | Apium graveolens L. var. dulce |
C67 | Jiazhouwang | Celery | Apium graveolens L. var. dulce |
C87 | Ventura | Celery | Apium graveolens L. var. dulce |
C114 | Huanghou | Celery | Apium graveolens L. var. dulce |
C62 | Introduced from USA(fertile) | Celery | Apium graveolens L. var. dulce |
C63 | Introduced from USA(sterile ) | Celery | Apium graveolens L. var. dulce |
C65 | - | Celery | Apium graveolens L. var. dulce |
C68 | Jiahuang | Celery | Apium graveolens L. var. dulce |
C74 | Dore Golden Spartan | Celery | Apium graveolens L. var. dulce |
C79 | Kangnaier | Celery | Apium graveolens L. var. dulce |
C83 | Bailixiqin | Celery | Apium graveolens L. var. dulce |
C89 | TU52-75 | Celery | Apium graveolens L. var. dulce |
C97 | Kaifengbolicui | Celery | Apium graveolens L. var. dulce |
C118 | Guihe | Celery | Apium graveolens L. var. dulce |
C121 | Qianfang | Celery | Apium graveolens L. var. dulce |
C123 | Ventura (yellow mutant) | Celery | Apium graveolens L. var. dulce |
C58 | Majiagouqincai | Middle type | Apium graveolens L. var. dulce |
C99 | Jinnanhuangxinqin | Middle type | Apium graveolens L. var. dulce |
C159 | Jinnanshiqin | Middle type | Apium graveolens L. var. dulce |
C163 | - | Celeriac | Apium graveolens L. var. rapaceum |
C130 | - | Wild | Apium chilanse |
2.1. Marker Development
Primer Name | Forward Primer | Reverse Primer |
---|---|---|
Fn1 | GCGCTTGGTGTATCTCCACT | AGTGCGTCGAAATATCGCTT |
Fn2 | GCTTCCGCTGTGTATTTTGA | GGGAAGAAACTGCAACTTGG |
Fn3 | TGAGCTCCACCAACTGACAC | GCATGAGCAGTTCCAAGACA |
Fn4 | AATTTACCGCTCTTAGCCTCG | ATAGGCAGAATTTGCGACGA |
Fn5 | TGAAACCCAAGATCACCCAT | TCATATTGACAGGCAACCGA |
Fn6 | CCAATCTGGGACTGTGTAAGC | TTCCTGGAGGTGAAGGACTG |
Fn7 | TGGTGTTGCAGTGTGAATCC | ACCGAAGCATCCTTGAACAG |
Fn8 | TGGTGTTGCAGTGTGAATCC | ACCGAAGCATCCTTGAACAG |
Fn9 | CATAGGCTAACGCAGCTTCC | AGTACTCCTTCAGCCGACGA |
Fn10 | CAGGAGGCTGCAATAACACA | GAGTCGCCGGAATATCAAGA |
Fn11 | CACACAGACGACTGCTGCTT | ACCATGCATGCTCAACTGAT |
Fn12 | CACACAGACGACTGCTGCTT | ACCATGCATGCTCAACTGAT |
Fn13 | CGATCAGGGTACTTGGCAAT | TTTCTATATCCGTCTCATTTCTGTT |
Fn14 | AACCCTAGCTGCCTCTCTCC | CCATGCCACGAATAGCATTA |
Fn15 | TGTGTTCTCGCATCTCCAAC | CCAATCTCAACATCGCACAG |
Fn16 | GTTGGTCAATGCTGCTTCCT | TGTGCCAGGGATACCTTCTC |
Fn17 | TCACTCACTCCCTTGAGCCT | TGAATCAACACCGTCCATTG |
Fn18 | CAACCTGAACATCGTTGGTG | TCAACTTGATCTCACGGCAG |
Fn19 | CTCATACGGTCCAGATCGGT | ATGTCCTGGTGAAGGAGGTG |
Fn20 | GAGATTGCGATAATGGTGGC | CGCATCACATCACTTAACGG |
Fn21 | CTGCTCTGAAAGGCTCTGCT | ACAGCTGACATCCTTACCGC |
Fn22 | TTCACTTGTTCAGCGAGACG | CCTAACCCTAGCTCGTCGTG |
Fn23 | TCCCATCTCCAATTCCAATC | TTCCTTGCAAGACCATAGGC |
Fn24 | CATGTCACTGTCGAAGCACC | TGACAATTGCCATTCTCCTG |
Fn25 | GCCTGAGCATCATAAGAGCC | TATTCACCTTCGTATCCGGC |
Fn26 | TGTTCCATTATGTGTTGCAGTG | GCGAGATGATGTCAGAACGA |
Fn27 | ACCATGTCCACCACCTTCTC | GCTGGTTATGGTGGTGCTG |
Fn28 | ATCGCCAACACCTTCTCAAG | AAGGGTGATTCTGATGGTGC |
Fn29 | CATATCCAGCACCTCCACCT | TCCAATGGCAACTCACAGAG |
Fn30 | CTCATTCTTCTTCTTGGCCG | TGCTGAAACGCTACCTCCTT |
Fn31 | ACATGGAATCTTTCACCTTCA | CATGGCCTAGGAGGAGACAA |
Fn32 | TTCCTGTCCAGCAGTATCCC | GAATTGAGGGGTGAAAAAGC |
Fn33 | AAATGAGGTGGTGGTGGAAG | CAATGGGTATGGAATCAGGC |
Fn34 | AGTACGGTGTCTACGACGGG | CCTCCACCATGATTACCACC |
Fn35 | CATACTTCTTTGGGGGCTCA | ACACAAACTTTCGGCCAGAT |
Fn36 | AAGGTCAAGGTCCTGTGGTG | GGTTTAGGCCTCCAATAGCC |
Fn37 | ACAGTACGTGTCTCCCCCTG | AACAACCCTATGATGGCTGC |
Fn38 | GTTTGAGCCTCCGCTTACAG | TGCCAGTGACACTCTTCACC |
Fn39 | GTGACGAAGGAATTGACCGT | ATTTGTTGTCGGGTTCCAAA |
Fn40 | CTGGCACTTGTACGAAACCA | TATGGGCTGTTGATGACAGG |
Fn41 | TTCAACCCAGACTTCAACCC | GCAGCCTTCAAATCCAGTTC |
Fn42 | CCCAGCCCTATCAATCTTCA | CCCCTGCCAAGTCTGTTAAT |
Fn43 | GAGACAGAGACCATGGGGAA | CGGTTTCGGTTTCGATTTTA |
Fn44 | TCTTGTCCATTAAAAATGTACCCA | TGCGCATAATGAAAGGATCA |
Fn45 | AGCAGCACAACAACACTTGG | TTAGGGTCTCTTGTCGAGGC |
Fn46 | GCAAGTTACCACCCCAGAAA | CCTTTTTCAAAAGCTTCCCC |
Fn47 | AATTGGCCAGAGCAGAGAAA | TCCTTTATCCCTGACCATGC |
Fn48 | AATGAGGTTGTTTTTCCCCC | GTAAAGGCCCAAACTCCTCC |
Fn49 | ACAACAATTTCAGGGCCAAG | TCTTGATATCGGCTTCCTGG |
Fn50 | ACATTTGTTGCTAGGGTGGC | GCACGAATAGCCGTCCTAAA |
Fn51 | TCCAATCTCCGGAGCAATAC | GCGGTGGACGAGTAAAAGAG |
Fn52 | AAACCACCAAACAAGGTGCT | TGAAGTGGAGGAGCAGGAGT |
Fn53 | ATTCCCAGATGGCTGCATAG | AATTCCAGCAAGCTCAAGGA |
Fn54 | CCCTCTCCCTATCTTCCTCG | TGAGATTGACTCGGTTGCTG |
Fn55 | AAAGAAGAAACGGGGATGCT | AACAGCAAGCAGTTCAGTAGTCA |
Fn56 | CTACACCGCCAATTCAACCT | TAAGCATACACCCCCTCACC |
Fn57 | TAATGGTGGAAAGAAAGGCG | GGCATACCACTCATTTGGCT |
Fn58 | CCTAGGCGAACTCTCCTCCT | GGGAATGATCCTCCTTCCAT |
Fn59 | AGAGGGATAAAATCCCGGTG | TGGAAATGCAAAAGAAGCAA |
Fn60 | CCACCACCACAACTACAACG | AAGCCGAGTAATGCTGGAAA |
Fn61 | AGGGTTCGTCGGTTGTAGTG | TCCCCATGTCTATTCTTCGC |
Fn62 | TCTCGACGAGTTTTCACCTG | GGTCTCTTTGGTGCCATTGT |
Fn63 | TCAATTAGATCCAGACGCCC | TCTTCTGCCTCCTCTTCGAG |
Fn64 | CTGTTTTCTCCCTCGTCTGC | CCCCATCTCTGCTATAGCTCC |
Fn65 | CTTTCTGGGTAAGAAGGCCC | CCAACCCACAACGTCTTACC |
Fn66 | TCATTGGACAAACAGGACGA | CTGTTTGGCGCTCAATTTTT |
Fn67 | TACATTTGTGGGATTGGGGT | CCGCCAAAACATTGACAGTA |
Fn68 | TCAGCTAAGCCACCCTGATT | GTTGCTGCTGGAGAAAGGAG |
Fn69 | TCCATGAATCTTTCAAGCCC | TCCAAAGTCCAATCCCATTT |
Fn70 | TAACTGAGTCGGTTGGGTCC | CCTCTCTTTTACCAGCCAAGC |
Fn71 | TCAACACTCAATTTAAACACCCA | TGATAACGATCGTGACGGAA |
Fn72 | CATCAACACTCGAAATCGAAA | CAAGATGCTTGTTATCCTTGCT |
Fn73 | GGATCGGAGGAAGGAAGAAA | GGAGGTGGAGGAGGAGAGTT |
Fn74 | CCCCCAAACAATAAGTATCCC | TTGGAACTTTTTGTGTCCATTG |
Fn75 | GCCAGCAGTGTCCCATATTT | GCCCGGAAAAATAACAATCA |
Fn76 | TTCTACCACTTTCCTTGAATCC | CAGGAGCAGTCTCGATTTCC |
Fn77 | GGTGTATTTGAATATTAACACCTTTCG | AGAGATGGTGGTCTTGTGGG |
Fn78 | ACAAGCCCCCTCTACTTTGG | ATGTTGCCAGTTCAGGTTCC |
Fn79 | TGGGACCCATTTCTTGATTT | AAAATTGCTCCGATTTGTGC |
Fn80 | CCAGGTAAGCCCAGTCTCAA | TTTTTCTCAATTAAAACTTGCTCATTT |
Fn81 | AATCCTTGAACTAACCGGGG | CTCTTCGCCACCAGATTCAT |
Fn82 | GGACGCCCAAGAGAGGTAGT | AGTGGTCTCGACATTTTCCC |
Fn83 | CCACACCTTGATCGTTGAGA | TTGCTTCTTCCGGCTCTTTA |
Fn84 | GTTACTTGACGGCACCGTTT | ATCAGTTCTTCATCCGTGGG |
Fn85 | TCACCCTCTCATCCACATCA | GCAGTGGGTGGATCTAGGAA |
Fn86 | TCAAATGGACGACGAATCAA | TGCAATGATTTATCCCCCTC |
Fn87 | CACACACAGGACACACATATTTC | GTAAAGCCGTCTTGGACGAG |
Fn88 | CGGCATCTTCTCTTCCTCAC | TGTTTGGATCTTTTCTGTTTTCA |
Fn89 | CAGAAGCGGCTCCTTCTCTA | CCCATTTGAGCTTCACCACT |
Fn90 | CTAAACGACGCCGTTACCAT | GCTTCTCTCCGCCTTGTATG |
Fn91 | GGCATACATCGGACGCTAAT | TTGACCCTTTATCTCAATACACACA |
Fn92 | CCCTCTCTCTCCCTCCTGTC | ACGATTAGCCATTGGTGAGC |
Fn93 | TGTGTGCTGATTTGAAACCC | ACCGACACTCCACCTTCATC |
Fn94 | CACCTCTGCTTTCACGGAAC | GTCCAAGAGTGGTCCTCACC |
Fn95 | ATGGTAACACCACCCTGGAA | GCTTCAACCAGGCAAAGACT |
Fn96 | TACTTACACCCCTCCCTCCC | TGCAGCACAAGGGATTCATA |
Fn97 | AAGAGCGATCAAGAACAGGG | TCCCATCTCTCTCCCTCGTA |
Fn98 | TGCGAACAATACAGTCCCAA | CAGATCCAAACACAGAATTAGCA |
Fn99 | GAAGAAAGAGGAGAAGGCCG | TCTCGAAACCACCCATCTTC |
Fn100 | GCGATCCCTAATCAATCCAA | CTTTGAGAGTTACGACGGGC |
Fn101 | TCAATGGTGTAGAACCAGAACAA | CCCAGATGCTTAAAAGAACCA |
Fn102 | ACAGGAGGCACTGGTCTCAC | CATTAAAATCCCACAAAAACTTCA |
Fn103 | TCCCATTCCATTTCAACCTC | AGAGGTGTGGGGAGATTGTG |
Fn104 | GCGGGGACACTCCACTACT | TTGATCATCAGCAGACTGGC |
Fn105 | CAAAAATTTAACCCCATACCC | ACATGTACGGACGTTGTGGA |
Fn106 | GCTTTGCACACACACACACA | CTTTCCCTCGACCTCATCCT |
Fn107 | GATTTTTCCGATGCAGCACT | GGCATGCACCAAACGTTATC |
Fn108 | GAGGAGGCTGTTACGTGGAG | TCCCTTTTCTCACTCCATTCC |
Fn109 | GTAGAAGGCCTGCAGATGCT | GTCTTTGCTTCTCCTCACCG |
Fn110 | GCACCAGCAAGAGGAGACTT | TTGTTGCTTGCCAGTGAAAC |
Fn111 | AAGCGAGTAGCTGAAAGGCA | CACTACCACCTCCGATTGCT |
Fn112 | CCAAGCTTCGACCATTGTTT | TTGTACATCGGTGAAACGGA |
Fn113 | AGCAGAAAGGCGTTCCACTA | GTTGAGCCCTTCCTGCATAA |
Fn114 | CGCCCTCTTCCTTTATCTCC | CACTTAGGTTTACCGCTGCC |
Fn115 | CACGTTTGGTGACATTCCAG | ACAATTATCTCCTTCCGCCC |
Fn116 | TCCTCTCCTCACCAAACCAC | CACAACCCTTCAACATCACG |
Fn117 | GTGGTTGGTGGGGATCATAG | GCCCAAAGTTCTTCCAAACA |
Fn118 | CAATCATATTAATCATCCCCAAA | GAGTTGGTCTGCAGGAGGAG |
Fn119 | TAAGATGCATGAGGCACGAC | GACTTTGATGCGCACTTTCA |
Fn120 | TGATTTGTGCACCAAAAGGA | GGAGAGTCGACCGATTCAGA |
Fn121 | AACTCAGCAACCGGAATCAC | ATACGTAGACGCATCGGAGG |
Fn122 | GCACACAATAAGCCTCCCAT | CACATGCTACAAAACAGGCG |
Fn123 | CCACTGGACATTTCTTGGCT | TTTACAAGCCCCAACAGAGC |
Fn124 | CTGGAACCGGAGTAGGTGAA | AACAGCCTTTACCCTTCATCA |
Fn125 | ATCTGCCTGTAGCCGAACAG | CTCTTAGTTGGCGCTGCTCT |
Fn126 | ATAATTTGCCCAACGCTCTG | CTCTCTTGAAAAACACGGGC |
Fn127 | CAACACAAACACCAAAACCCT | CGTGCCTCATTGGGTTCTAT |
Fn128 | GTTGTACTTGGTGCGGAGGT | CAAAATTCCAAAAGCCCAAA |
Fn129 | TCTTTCGATTTGGATTTGGG | TAGAGCTCTCGGCCTCTTCA |
Fn130 | TTGGTGCCATTGTTGTTGTT | AACGCCTTTCTTCCCAATTT |
Fn131 | CACCGCGATTCTTCTCTCTC | CGACATCGTCTCTCTCCCTC |
Fn132 | TTCTTTTTCTGTTCCGCCC | CCGCCGTTAGAGACAAACTC |
Fn133 | ATTGAAACCCCACCACTGAA | AACGGCCAGAAAAAGCTGTA |
Fn134 | TGGTTGGGGGAGAATTGTAA | TGAGTTTGCCACAACTGACA |
Fn135 | TCCCGATAACAAGAGAGAGACT | TGGAGATGAACAAGGGAAGG |
Fn136 | AGTCCTCAGTTCTCCTGGCA | CAGAATGGTGATGCTGATGG |
Fn137 | CCAGGACATACATACGTTCTCAA | GACGGACTTAGCCCCCTTAT |
Fn138 | TAGCTGCGGTTGATTCAGTG | ATTATCAGCGGAAGGCACAC |
Fn139 | TGCACCACCAAAAACACCTA | GAGGAGGGGTTGAGTGATGA |
Fn140 | TCACCACCCCTAATTACCGA | AGATAAACCGGGGAGCTTGT |
2.2. Marker Informative Analysis of Accessions
2.3. Analysis of Molecular Variance (AMOVA)
Source of variation | Percentage of variation | p-value |
---|---|---|
Among horticultural types | 22% | 0.001 |
Among Individuals | 43% | 0.001 |
Within Individuals | 35% |
Horticultural type | Local celery | Celery | Middle type |
---|---|---|---|
Local celery | - | ||
Celery | 0.231 | - | |
Middle type | 0.086 | 0.261 | - |
2.4. PCA Analysis
2.5. Genetic Distance Analysis
2.6. Cluster and Population Structure Analysis
2.7. Genotypic Diversity
3. Experimental
3.1. Plant Materials and DNA Isolation
3.2. Development of Genic SSR Markers and Genotyping with Markers
3.3. Analysis of Marker Polymorphism and Genetic Heterozygosity
3.4. AMOVA and PCA Analysis
3.5. Genetic Diversity and Population Structure Analysis
3.6. Identification of Genotypes
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Fu, N.; Wang, P.-Y.; Liu, X.-D.; Shen, H.-l. Use of EST-SSR Markers for Evaluating Genetic Diversity and Fingerprinting Celery (Apium graveolens L.) Cultivars. Molecules 2014, 19, 1939-1955. https://doi.org/10.3390/molecules19021939
Fu N, Wang P-Y, Liu X-D, Shen H-l. Use of EST-SSR Markers for Evaluating Genetic Diversity and Fingerprinting Celery (Apium graveolens L.) Cultivars. Molecules. 2014; 19(2):1939-1955. https://doi.org/10.3390/molecules19021939
Chicago/Turabian StyleFu, Nan, Ping-Yong Wang, Xiao-Dan Liu, and Huo-lin Shen. 2014. "Use of EST-SSR Markers for Evaluating Genetic Diversity and Fingerprinting Celery (Apium graveolens L.) Cultivars" Molecules 19, no. 2: 1939-1955. https://doi.org/10.3390/molecules19021939