Study on sequence variation in rc gene of vietnamese local pigmented rice

VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY -oOo - GRADUATION THESIS STUDY ON SEQUENCE VARIATION IN Rc GENE OF VIETNAMESE LOCAL PIGMENTED RICE HANOI, 2021 VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY -oOo - GRADUATION THESIS STUDY ON SEQUENCE VARIATION IN Rc GENE OF VIETNAMESE LOCAL PIGMENTED RICE Student : Nguyen Thi Yen Chi Class : K61CNSHE Faculty : Biotechnology Supervisor : Nguyen Quoc Trung, MSc Chu Duc Ha, PhD HANOI, 2021 COMMITMENT This thesis is composed of my original works, and contains no material previously published or written by another person Hanoi, 30th January, 2021 Student Nguyen Thi Yen Chi i ACKNOWLEGEMENTS During the process of studying, researching and completing the thesis, I have received the help of many individual First and foremost, I would like to express my gratitude to my supervisor MSc Nguyen Quoc Trung, Department of Biology, Vietnam National University of Agriculture, and Dr Chu Duc Ha, Agricultural Genetics Institude, for their patience, motivation, and immense knowledge Their enthusiasm guidance helped me all the time of researching and writing of this thesis Besides my supervisors, I would like to thank guides and people working in Crop Research and Development Institute for sharing their experiences, time, and commitment during my work to prepare my materials as rice samples I also would like to offer all other friends, who also thesis in Center of International Plant Research Vietnam and Japan for their kindly help, care and lift me up all the trouble in the time doing experiments My sincere thanks also go to teachers in the Faculty of Biotechnology, Vietnam National University of Agriculture, whose encouragement, guidance, and inspiration supported me during my studying time at higher education Besides that, I am thankful to my dear classmates, class K61CNSHE, for all the time we had in the last four years Last but not least, I am grateful to my family and my friends for supporting me spiritually throughout writing this thesis and my life in general I sincerely thank you! Hanoi, 30th January, 2021 Student Nguyen Thi Yen Chi ii TABLE OF CONTENTS COMMITMENT i ACKNOWLEGEMENTS ii TABLE OF CONTENTS iii LIST OF TABLES v LIST OF FIGURES vi LIST OF ABBREVIATIONS vii ABSTRACT ix CHAPTER INTRODUCTION 1.1 Introduction 1.2 Objectives 1.3 Requirements CHAPTER LITERATURE REVIEW 2.1 Evolution and domestication of rice cultivar 2.2 Overview of theRc gene in rice 2.2.1 Anthocyanin 2.2.2 Studies on genes controlling anthocyanin synthesis 12 2.2.3 Studies on Rc gene 16 CHAPTER MATERIAL AND METHODS 18 3.1 Materials 18 3.2 Field layout 18 3.2.1 Experimental layout 18 3.2.2 Cultivation techniques 18 3.2.3 Methodology 19 CHAPTER RESULTS AND DISCUSSION 24 4.1 Results of cloning and amplification of Rc gene fragments 24 4.1.1 Design the primers for sequencing 24 4.1.2 Optimization of PCR for cloning fragments 26 iii 4.2 Results of sequencing Rc gene fragments 31 4.3 Alignments of Rc gene 31 4.3.1 Rc gene in Nep cam dang 31 4.3.2 Rc gene in Nep cam (Tuyen Quang) 33 4.3.3 Rc gene in Nep cam den 35 4.3.4 Rc gen in Nep cam (Hoa Binh) 36 4.4 Identification of 14 nucleotide deltion in seventh exon 38 4.5 Sequence variations in Rc gene (SNP) 39 4.6 Phylogenetic tree 40 CHAPTER CONCLUSIONS AND SUGGESTIONS 41 5.1 Conclusion 41 5.2 Suggestion 41 REFERENTS 42 iv LIST OF TABLES Table Example of rice genes involved in flavonoid biosynthesis (Furukawa et al., 2006) 13 Table Information of four local pigmented rice varieties used 18 in this study 18 Table Component of the PCR reation 21 Table The Annealing temperature of each primer 21 Table The optimal annealing temperature in PCR for each reasponsible primers 26 Table Total 17 fragments were sequenced 31 Table Collinear analysis of the seventh exon's partial sequence of Rc gene for local pigmented rice varieties 39 Table The number of SNP identified by comparising the Nep cam dang variety with varieties ( O sativa indica and O.sativa japonica) 39 v LIST OF FIGURES Figure1 Basic anthocyanin structure Figure2 Two-dimensional structure of flavylium ion 10 Figure Metabolic pathways for the synthesis of pro‐anthocyanidins and anthocyanins 11 Figure Rc allele phenotypes (Sweeney, et al., 2006) 14 Figure 5, 6, The primers were designed locating overlap along Rc gene sequence 25 Figure PCR product electrophoresis before and after determining the optimal annealing temperature 28 Figure 9,10 Some fragments with single bands 29 Figure 11 The product electrophoresis is Rc_8 fragment 30 Figure 12 Phylogenetic tree of 'Nep cam dang 2' variety 40 vi LIST OF ABBREVIATIONS bHLH basic helix-loop-helix BC Before Century AD Anno Domini QTL Quantitative trait locus FNP Functional nucleotide polymorphism ABP Anthocyanins biosynthesis pathway DFR Dihydroflavonol‐4‐reductase PCR Polymerase chain reaction vii LIST OF APPENDIX Appendix table Sequences of 17 fragments in Nep cam dang 45 Appendix table Sequences of 17 fragments in Nep cam (Tuyen Quang) 48 Appendix table Sequences of 17 fragments in Nep cam den 51 Appendix table Sequences of 17 fragments in Nep cam (Hoa Binh) 55 viii 26 Sakamoto, W., Ohmori, T., Kageyama, K., Miyazaki, C., Saito, A., Murata, M., Noda, K and Maekawa, M (2001) The Purple leaf (Pl) locus of rice: the Pl(w) allele has a complex organization andincludes two genes encoding basic helix-loop-helix proteins involvedin anthocyanin biosynthesis.Plant Cell Physiol.42, 982–991 27 Saitoh, K., Onishi, K., Mikami, I., Thidar, K and Sano, Y (2004) Allelic diversification at the C (OsC1) locus of wild and cultivated rice: nucleotide changes associated with phenotypes Genetics, 168, 997–1007 28 Kato, S., & Ishikawa, J (1921) On the ingeritance of the pigment of red rice Jap J Genet, 1, 1-7 29 Tan, L., X Li, F Liu, X Sun, C Li, Z Zhu, Y Fu, H Cai, X Wang, D Xie and C Sun (2008) "Control of a key transition from prostrate to erect growth in rice domestication." Nat Genet40(11): 1360-1364 30 Reddy, V.S., Scheffler, B E., Wienand, U., Wessler, S R., Reddy, A R (1998) Cloning and characterization of the rice homologue of maize C1 anthocyanin regulatory gene Plant Mol Biol 36, 497– 498 44 APPENDIX Appendix table Sequences of 17 fragments in Nep cam dang Name Rc gene Sequence of Rc gene fragment fragment AF1 AF2 AF3 AF4 AF5 RC_1 AGGGCTTTGCTACTAGCTCCTCACTCACTCGCCTTTACTTATCGATCTCGATCATCCACGAGCTAG CTAGCAGTGCTCGCCATGGCCGGCGGCGAGGCGCATGCGGCGCTGCAGGCGGTGGCGCAGAGCCTC CGGTGGACCTACAGCCTCCTCTGGCAGCTCTGCCCCCACCAAGGGTACCTACCCTACCTACCTACG ACACGATGCACAGTGTTCATCCATGGCCGGCCATGGCGGATCGTCGTCGTTGTCGATGATCATCGA AGGAAGCTAGAGGATATGGCTCAATACTTTGATAATATATATACTGATCTCTCCGTACAACAAAAA TATAAAAATTCTAGCTAGTATCGAATGAGACATATGCTATGCTAGTACTACGAATCTAAAAAGATG TACATATTTTGATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGACGGATGTAATAA TTCTGAATTTAGTTGTGATCGCATGGCATA RC_2 TCCGATGAGACTATGGCTATGCTAGTACTACGAATCTAAAAAGATGTACATATTTTGATTCGTATT ATTAGGATATATCACGAGTTTTTATATTTTGAGACGGATGTAATAATTCTGAATTTAGTTGTGATC ACATGGCATGCAGGAGCTCGCTGGTGTGGGGGGAGGGGCACTACAACGGCGCCGTCAAGACGCGGA AGTCGACGGTGATGCAGCCGCCGCCGGCGGAGGAGGAGGACGACGCCGACCACGCGGCGCGCCACC GGAGCCGGCAGCTGAGGGAGCTCTACGACTGGCTGCAGCAGGCCGGGGAGAACTCCAGCGGCGGCG TGCAGACGTCGTCGACGACGGCGAGCCGGCGGCCGGGGGCGGCTCTGTCGCCGGAGGACCTGACGG AGACGGAGTGGTTCTTCCTCATGTCGGCATCCTACTCCTTCCCTCCCGGCATCGGGTATATAATAA AAAAGAAAGATATAAATATTAAATGCATGCATGCATCACATTATACCTCTCTTTTTGTTA RC_3 GGGGGGGAATCCAAGCGGCGGCGTGCAGACGTCGTCGACGACGGCGAGCCGGCGGCCGGGGGCGGC TCTGTCGCCGGAGGACCTGACGGAGACGGAGTGGTTCTTCCTCATGTCGGCATCCTACTCCTTCCC TCCCGGCATCGGGTATATAATAAAAAATATAGATATAAATATTTAAGCATGCATGCATAAATTAAA CCACACTTCTTGTTACGTGTTCTTGGCAAAATGATGAACAATTACCACTAATTAATTGGAGCCAGA AACCCTAAAGATTTACCCACCTGGTTAATTAATCGGTGTGTTGATCCACGCATGCATGCATGCAGA AAATCAAGATCAGGATAGCTCCTTTTCTTTTGCAGGTTAATTAGCTAGATCTTCACGTATAATTAG CTAGCTAGATTTTAAAATATAATTTATTCAATTTGATTTATGATTTTTATTTTTTATTTCAAATAG ATACAACTGTATACAAAATTATATTTTGGTACATACCTCCGATCCAACTACATCAGAGGTAAAAAA AAAATTAAACCGTTGGAATTGATTAGAACAAGATCGTGCGGCCAGTCGTGAGAGCTCCCGTCACAA RC_4 TTCCCAAACTTCCGTATAATTACGCTCGCTTGATTTGCAATCATAATTCTACGTCATTTCTATTGA TGCTTTTTATTTCTTATTTCGCATAGATACAACTGCTATACAAGATTTTATTTTGGTACATACCTC CGATCCAACTACATCAGAGGTAAAAAAAAAATTAAACCGTTGGAATTGATTAGAACGGAATCGTGC GGACAAATTATATCATAACTAACTTTTCTGATTCTCTAAAGCATAGAGATGTATATATACATCGTA TTATAAGGCTCTATATTTCCTGATTAACACTAGATGCATATATAATTTTGATAGTAAAAATATACT TTTGATAGGCTCTAAAGAAAAACTTAATAACATGTACTCCCTCCATATACTTTTGATAGTCATATT TCATCTTGACCCACAGATCAAGTATAGGTAATTCTACTTATCATCCATTTAACCACGCTACTAGTT ATTCCTCATAAACAAGCGATTCATTAATATTTACATTTCTCGATGCTTGTGTAGCCAATATTGTGT GGAAGAATGGAATGTCATTAAGAGGATAGGTTGTTGGATTGAAATATACCAAAG RC_5 TAGGAAGGATGTCATTAAGAGGATAGGTTGTTGGATTGAAATATGCCTATCAAAAATAAATTTTTA GATTTGAAAATATGCCTATCAAAAGTAGATGGAGGGAGTATTAATTAATGTGAATTTCCAATCCTA CTGTTGTGATATTAGGCTTTGTACCTTCTTGTCCAGGAGGTATATATATGGCTCTTTTAAGGATGG GAGAAAATATCATCTTTAATACAACTATATATGGCTTTTGTTTGATAAATACAACTTTTATTTTGT ATGAATACAAATATATTGATAAATATCCACCATTATAATCCTAACCCATTAGGATCATATGGTGTA TATTTTTTTAACTATTTGTTTTTTATAAATTAATATTAAGAGATCACAATAAAAATATAGTATTAT GAAAGTACTCTTAACAACATATCCAATGATAAAATTATTATTATTACAAAATATAGTGGTCAAATT GTATAGAATTCAATAGCCTGATTTTATGACGTCAAGTAAATTAAATAAAGAATGAAGGTAGTGCTA AGTGGGATCAAAA 45 AF6 AF7 AF9 AF10 AF11 AF12 RC_6 TGAGGGTAGTGCTAGAGTGATCAACAATATCTCTCCTAAAATATGTCCTATAAGTTTTACTCCATA AATCCAAGGGTCAAAAGTTGTTGGGTTATTTTTTTAGATAATAACATACTACCCCTTTTCAAAATG TATGATTCTATTGACTTTTTGCACAACATTTAACCATTTGTCATATTAAAAATTAGTATAAACATC TAAAAATATAAGTTACAATTATATTTTATTTGATGATAAAACAACTCACAACAAAATAAATAATAT TTATATAATCTTTTTGGAATAAAACGAATGATCAAACATTATTCAAAAAGTCAATGGTATAGTACG TTTTGAAATTGATAGACTATGAGAGCAAAATTTTGAGATAACATGGAAAATTATCCTCTTAGACAT TGCACTGTGTAATAATTAATAATAATGAATGAAAGGCTAAGACTTTTCTTCCACCTTATATAAGTG GTTGAATATATAGCAATCACATCATTACATGATTTTGTAACCAACCGTCTCTATAGCTCCGAATAC CAGTGCAA RC_7 AATGGTGATTAGCAATCACATCATTACATGATTTTGTAACCAACCGTCTCTATAGCTCCGATACAG TGCTAGTTTCACATCGTAATAATTAAAGAGTATAATAATAAATCGAGGTGTACTTCTCATCGATGA AGTGATGTGCCGCTTAGCTAAATTAAACTCGTATGCAAAAATCAGTATATGTCCGGTTAATTTCTA AGAGAGAGATTGAGAGAGAATAATTGCGCCCCTCCAAATCCCCCTCTTGGACGTTAGGGAGCTATA TAGACGGTATTGCTAAGTGCGATGTGTACATAACGTACCTGTCGGAGGAACATTTCTCATCCAAAT TAAGTAGTAATGGATGGCATGAAATCCATTTTTGTATTATGCTTGTACTACGAATCTGACAAGATG TACATAATTTGATTCTGATTATTACGATATATCATGAGCATATTATATCTTGACACGGATGTTATA ATACAGAATATAGTTGTGACATA RC_9 CGTGCTTTCATTGTCATGAGGAGAGAACTTGTTAAGTTGCACACATGTGTGCTTTGCATGTCTTCA GGTTACCTGGAAGGGCATTTGCAAGGAGAGGCCATGTATGGCTCACTGGAGCAAATGAAGTTGACA GCAAAGTATTCCTAAGAGCAATTCTTGCCAAGGTTCAGCCATCACCTTCTCTTACCTATTTTTCAC TCTGAATGCCAACAGTGCTTTGCACATTGTAGTCTGTTTGCAGACTGCAAATGATGACCATAATCA GATCAGAAAATAAAATAATATTATATACTTTTTGAGCCAGCTAGCAAGAATATGTAACAATAATTC TCCTTTTTTTTTCTTGTTCTTTTCCCTGATGTGGTGCATAACAAATAACCAAACTGATGAATGGCA AAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTACTATGAAATTTTGTGGCTT TTGTTCATCTTTGGATGCAGTGG RC_10 AAACGGAGAAGGCAGAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTACTATG AAATTTTGTGGCTTTTGTTCATCTTTGGATGCAGTGGCCATTATCTAAAAACTATGAATTTCCAGA CTGCAGTTTTTATCTAATTTTGTGACTTTGTACATCAGACAGTTGTGTGCATTCCTGTTGTCGATG GCGTCCTGGAAATTGGAACTACGGAAAAGGTGATTTCGTATATTATCAGCTGACAATCTAATTATA TGGGCCATATAATTAAGTATAAATCAAAATACCTCATAATATATTATAAAGTATCTAATGTGATTA TGTGAATATTGGCTATTTCAATGTAATTTGATATATGAAACTGATAATCCTCTGAAACTCCGTAAG GATCAAACTAATCAAAATGTATATATTTTCAAGGTGGAGGAAGATATGGGCCTGATTCAGTATGCA AGGGGCATCTTCATGGATCAACATGGCATCCACATGAACCCTAACCCTCAA RC_11 TCATCCACATGAAGCCTACCCTCTCACAGCACTCAACATCCAACCCAGTCACCCACTGTACTCATC AGCATCCAATCCAGGTTCAGATGCAACTAGGTATCACCAGCCAAACAAAGTTTGATTATTCAGATG AGCTCAATGCAGATGAGGAGAATGATGACACAGAAGAAGAGGGCATGTCAGGTTCAGACACTAACA ACACTGACACTGAAAGGAATTCAGGCCAGCTGCAACTTCAAATGCAAGACCAACTGAACATGGTGA GCAATGACCACCAGACAATACCAAATAATGCAGTTTCCAGTGAGCTAATGCAGTGTGAGATGTCAG AAGTGGTAAGAGATGGCTGCTCAAATAATATTTTAGAGGATGAAATCCAAATGCTGATGGATTGCC AAAACAGTAATTGTCAGTTAAATTTGCAAGGGCCAGATGAGCCTTGTCACTCTTGGCATTTTCTCT GCGAGGAGTTACAAAATGATTACCAGCCAGGTATTACATTTGAGAAGATAATCCTTCAAAAGCACC CCTTGTTCCATCCCTTCTCATTCAA RC_12 GTAAATGTATCCTATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATAGATATATATATATA GGATATCTATAGA 46 AF13 AF14 AF15 AF16 AF17 RC_13 AGAGATGTTCCAGTAGTCACTGCAGTTACAGGGGAGCAGAAACACCTGAATCAAGGGGCGGGAAAG GCGCAAGTGGATGCCATCCAAGGTGATTTCAGTGCCAACCATGTGCTGAAAGAGAGGAGAAGAAGA GAGAAGCTCAATGAGAAGTTCATAATTCTGCGATCTTTGGTACCTTTCATGACAAAGGTAATTAAG TACTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGCAAATAT GTAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAAAATAAA ACATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCACATATTT TAAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAAGCCTCT AACGAACATTGGAAGATCCTCACTAATGGCAGCATGA RC_14 CCCGAGGGAGCATGAATCTAGGGTTCACTATTTCGGAATGCAAATATGTTTTACCGGGCATCCGAT TTTTAAAAAATTCAGAATGAAGAAAATTGAATCTTTTTTATGGATTTGAATAAATCTTGATAAATT CGAAAAAATTTCCGAACTTTTGGCCAGAAGTGAATCCTACCCGTATCCACCGGTAATAAACCTAAA TTTTTGGGAGTAATGAATTAATGTTATATATAATCCATGAATTATATAGTTCCAAACTACTCCGTA ACAAATTTTCAGGAGTAGTGAAATTAATATTATTACAATCTCAGAAAAAAATGGCAGAAACAATTA ATCTGTTTTCAATTATTAATTAATTTGTTTTTGTGTCCAGATGGACAAGGCGTCGATACTAGGCGA CACGATCGAGTACGTGAAGCAGCTAAGGAACCGCATACAAGAGCTCGAGTCGTCGTCGTCGTCGTC ACGAGCAGCCGCCCGGGCGCCATCGGCGGCGGCCGCCGGGAGGCGGAGGAAGAGATCCGCCGCCGC CGCCACTGCCACGGCGGCGGAAGGGATGAGCAGCAGCAATGGCCGCAATGGCGGCGAGGCGGCGGA GGTGGTGCAGGTGTCCATCATCGAGAGCGACGCGCTGCTGGAGCTCCGGTGCGGTTGCGGCGGCGG CGGCGGCGGTGTGGTGCTGCTCCGGGTGATGCAGGCGATGCAGGAGCTCCAGCTGGAGGTCACCGC CGTCCAGGCCTCGTGCGCCGGTGGCGAGCTGCTCGCCGAGCTGCGCGCCAAGGTCGTCGTTATGAT CCTGATCTGCATGAAAATGCAGATGCAGATGCAAATGCAGAATTAAGCTTTCATTCTTGGCTCCTA TGAA RC_15 AAAAAAAAAATATATATATATAGTACCACTCTGTCTCTCATCTCTATATATATATCTATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATTTTTTTATATATATAATATATATCTATCTAT ATATATATATATATATATATATATATATATATATATATGTATATATATATATATATATATATATAT ATATATATATGTATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATTTATATTTATATATATATATATATATTTTTATTTTTTTATTTTTTTATTTTTAT TTTTTTTTATATTTATTTTTTTTTTTTTTTTTTTTTTTTATCTTTTTTTTTTTTTTTTTCTTTGTT TTTTATATGTATATATACTCTATATATATATATGTATATATATTCTATATATATAATATGTGTATC TCTCTATATATGTACTGTATACTGTATATGTATCTCGAGTGTATATGATGTGTGTCTATCTATGTA TTATATATATATATACACTCTCTCTATCTCTATATACTATATATATGTATACTATATTATATCTAT TATATATATTCTTTATATATTCTCTCTTTTCTCTATATATATATATCTATATTAGTCTGTCTATTC TGTTCTTACTATCGTCGTATATATGTGCTTATTGTATATACTAGTCTATACTATACTAATTC RC_16 GGTCTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGCAAATA TGTAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAAAATAA AACATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCACATATT TTAAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAAGCCTC TAACGAACATTGGAAGATCCTCACTAATGGCAGCATGAATCTAGGGTTCACTATTTCGGAATGCAA AATATGTTTTACCGGGCATCCGATTTTTAAAAAATTCAGAATGAAGAAAATTGAATCTTTTTTATG GATTTGAATAAATCTTGATAAATTCGAAAAAATTTCCGAACTTTTGGCCAGAAGTGAATCCTACCC GTATCCACCGGTAATAAACCTAAATTTTTGGGAGTAATGAATTAATGTTATATATAATCCATGAAT TATATAGTTCCAAACTACTCCGTAACAAATTTTCAGGAGTAGTGAAATTAATATTATTACAATCTC AGAAAAAAATGGCAGAAACAATTAATCTGTTTTCAATTATTAATTAATTTGTTTTTGTGTCCAGAT GGACAAGGCGTCGATACTAGGCGACACGATCGAGTACGTGAAGCAGCTAAGGAACCGCATACAAGA GCTCGAGTCGTCGTCCTCGTCGTCACGAGCAGCCGCCAGGGCGCCATCGGCCGAGGCCGCGAGGAA GGAGGAGGACGACATCCGCC RC_17 CATCGGCGGCGGACGCCGGGAGGCGGAGGAAGAGATCCGCCGCCGCCGCCACTGCCACGGCGGCGG AAGGGATGAGCAGCAGCAATGGCCGCAATGGCGGCGAGGCGGCGGAGGTGGTGCAGGTGTCCATCA TCGAGAGCGACGCGCTGCTGGAGCTCCGGTGCGGTTGCGGCGGCGGCGGCGGCGGTGTGGTGCTGC TCCGGGTGATGCAGGCGATGCAGGAGCTCCAGCTGGAGGTCACCGCCGTCCAGGCCTCGTGCGCCG GTGGCGAGCTGCTCGCCCAGCTGCCCGCCAAGGTCGTCCTTATGATCCTGATCTGCATGAAAATGC 47 AGATGCAAATGCAAATGCAGAATTAAGCTTTCATTCTTGCTCCTCTGAATTCTGAATTTATATATT CACCCTTCTTTCGATCTGCTCGTACGTTCGCTTCCCCTAGATTATGTACAAATTAACTGAATCTTT GAACTGAAAATAACTGAATCTTTTTCGTGTGTTTATGTGTGGGTGAATTGGTTGGCGCAGGCGAAG GGGAGGAAGAGGAGCAGCTTTGCTCACGTGATGAGGGGCATCCATCGCGTCCTCTCCCCATCTATA TCACTATTAAAT AF18 RC_18 TTCGGTTAATTATATAGAAAGTTCACATATATAAATCATGATAATTACCTTTTAGAAAATTCAAAC AAAGAATCGATATATATAGTTCTATCTATGCATCATTTTCCTAAATCACGATAATTACCTCAAGCT TGATAAAATGGCATTACATTACTATTATTATTACTAGAGTTTTTTCACTTCCCTGATCTAACCAAT TTGCCACGATGGTTAGATATATAGATGGTTCTAATTAATCTAATGTTAATGTACTCCACATATATG ATATATGTACATACATATCTAGTTTTAACATTTGCAAATGAATTGCAATCAGAATATATATATGCA TGCAGCACAGGTGCTTGATTGCAATTATATGATTATTACTCTCTCACATATTGATATGGTAAATGT GTATATATTTGTGGCGGGTTAATAACCATAATGGCAGGATGGGATTATAACATACCTCCAATTATC AAAATCAAACGTGGGATGAATCACCATCATCCTGCAAGTGAGCATTGTGGCCGGGTAATTATTAGA CGATCCTCGGCTCCAGAAGCATCCATCCCAGATGATGCGACGCTAAAC Appendix table Sequences of 17 fragments in Nep cam (Tuyen Quang) Name Rc gene Sequence of Rc gene fragment fragment AF1 AF2 AF3 RC_1 AGGACTTACTACTAGCTCCTCACTCACTCGCCTTTACTTATCGATCTCGATCATCCACGAGCTAGC TAGCAGTGCTCGCCATGGCCGGCGGCGAGGCGCATGCGGCGCTGCAGGCGGTGGCGCAGAGCCTCC GGTGGACCTACAGCCTCCTCTGGCAGCTCTGCCCCCACCAAGGGTACCTACCCTACCTACCTACGA CACGATGCACAGTGTTCATCCATGGCCGGCCATGGCGGATCGTCGTCGTTGTCGATGATCATCGAA GGAAGCTAGAGGATATGGCTCAATACTTTGATAATATATATACTGATCTCTCCGTACAACAAAAAT ATAAAAATTCTAGCTAGTATCGAATGAGACATATGCTATGCTAGTACTACGAATCTAAAAAGATGT ACATATTTTGATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGACGGATGTAATAAT TCTGAATTTAGTTGTGATCGCATGGCATAATCTCGCAGTATACTACCTATTTTTTTGCATCCTTGG AGTACGGCGAGGTCCGTATATATATGGTCATGGTGGTGAGCGGTGTACTCTGGCTTTTTTCGTTG RC_2 TCGGTTCGAATGAGACATACTGCTATGCTGTACTACGAATCTAAAAAGATGTAC ATATTTTGATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGACGG ATGTAATAATTCTGAATTTAGTTGTGATCGCATGGCATGCAGGAGCTCGCTGGT GTGGGGGGAGGGGCACTACAACGGCGCCGTCAAGACGCGGAAGTCGACGGTGAT GCAGCCGCCGCCGGCGGAGGAGGAGGACGACGCCGACCACGCGGCGCGCCACCG GAGCCGGCAGCTGAGGGAGCTCTACGACTGGCTGCAGCAGGCCGGGGAGAACTC CAGCGGCGGCGTGCAGACGTCGTCGACGACGGCGAGCCGGCGGCCGGGGGCGGC TCTGTCGCCGGAGGACCTGACGGAGACGGAGTGGTTCTTCCTCATGTCGGCATC CTACTCCTTCCCTCCCGGCATCGGGTATATAATAAAAAATATAGATATAAATAT TTAAGCATGCATGCATAAATTAAACCACACTTCTTGTTACGTGTTTTCTTGGCA A RC_3 AGGCGGGGGAATCCAGCGGCGGCGTGCAGACGTCGTCGACGACGGCGAGCCGGCGGCCGGGGGCGG CTCTGTCGCCGGAGGACCTGACGGAGACGGAGTGGTTCTTCCTCATGTCGGCATCCTACTCCTTCC CTCCCGGCATCGGGTATATAATAAAAAATATAGATATAAATATTTAAGCATGCATGCATAAATTAA ACCACACTTCTTGTTACGTGTTCTTGGCAAAATGATGAACAATTACCACTAATTAATTGGAGCCAG AAACCCTAAAGATTTACCCACCTGGTTAATTAATCGGTGTGTTGATCCACGCATGCATGCATGCAG AAAATCAAGATCAGGATAGCTCCTTTTCTTTTGCAGGTTAATTAGCTAGATCTTCACGTATAATTA GCTAGCTAGATTTTAAAATATAATTTATTCAATTTGATTTATGATTTTTATTTTTTATTTCAAATA GATACAACTGTATACAAAATTTTATTTTGGTACATACCTCCGATCCAACTACATCAGAGGTAAAAA AAAAATTAAACCGTTGGAATTGATTAGAACAAGATCGTGCGGACAGGCGGAGACTAG 48 AF4 AF5 AF6 AF7 AF9 AF10 RC_4 ATTGCTGATCTTCCGTATAATTAGCTAGCTAGATTTTAAAATATAATTTATTCAATTTGATTTATG ATTTTTATTTTTTATTTCAAATAGATACAACTGTATACAAAATTTTATTTTGGTACATACCTCCGA TCCAACTACATCAGAGGTAAAAAAAAAATTAAACCCGTGGAATTGGTTAAAACCAAAACCGGGGGG CCAATTTATTCCAAACCAACTTTTCCGAATCCCCAAAACCAAAAAAAGGAAAAAAACCTCCTAATA ATAAGGCCCTAATTTCCCGGATAACCCCAAAAGGCTAAATAAATTTGAAAAGCCAAAAAAACCTTT GAAAGGGCCCAAAAAAAAACTTAATAACCTGGTCCCCCCCCCAATACCTTTGAAAATCCAATTTCC TCCTGGCCCCCCAATCAAGGAAAAGTAATTCCACCTATCCTCCCTTTTAACCCCCCTCCAAGTTTT CCCCCAAAACAAACCATTCCTTAAAAATTTACCTTTCCCCCATGCCTGGGGAACCCAAATTGGGGG GAAAAAAGGAAAGGCCTTTAAAAGAAAGGGTGGTGGAATTAAAATATTGCCAAACA RC_5 CTTTTGAAGGATGTCATTAAGAGGATAGGTTGTTGGATTGAATATGCCTATCAAAAATAAATTTTT AGATTTGAAAATATGCCTATCAAAAGTAGATGGAGGGAGTATTAATTAATGTGAATTTCCAATCCT ACTGTTGTGATATTAGGCTTTGTACCTTCTTGTCCAGGAGGTATATATATGGCTCTTTTAAGGATG GGAGAAAATATCATCTTTAATACAACTATATATGGCTTTTGTTTGATAAATACAACTTTTATTTTG TATGAATACAAATATATTGATAAATATCCACCATTATAATCCTAACCCATTAGGATCATATGGTGT ATATTTTTTTAACTATTTGTTTTTTATAAATTAATATTAAGAGATCACAATAAAAATATAGTATTA TGAAAGTACTCTTAACAACATATCCAATGATAAAATTATTATTATTACAAAATATAGTGGTCAAAT TGTATAGAATTCAATAGCCTGATTTTATGACGTCAAGTAAATTAAATAAAGAATGAAGGTAGTGCT AGAGTGATCAAAA RC_6 ATAAAGAAGAAGGTAGTGCTAGAGTGATCAAACAATATCTCTCCTAAAATATGTCCTATAAGTTTT ACTCCATAAATCCAAGGGTCAAAAGTTGTTGGGTTATTTTTTTAGATAATAACATACTACCCCTTT TCAAAATGTATGATTCTATTGACTTTTTGCACAACATTTAACCATTTGTCATATTAAAAATTAGTA TAAACATCTAAAAATATAAGTTACAATTATATTTTATTTGATGATAAAACAACTCACAACAAAATA AATAATATTTATATAATCTTTTTGGAATAAAACGAATGATCAAACATTATTCAAAAAGTCAATGGT ATAGTACGTTTTGAAATTGATAGACTATGAGAGCAAAATTTTGAGATAACATGGAAAATTATCCTC TTAGACATTGCACTGTGTAATAATTAATAATAATGAATGAAAGGCTAAGACTTTTCTTCCACCTTA TATAAGTGGTTGAATATATAGCAATCACATCATTACATGATTTTGTAACCAACCGTCTCTATAGCT CCGATACAGTGCAAA RC_7 AATGGTGAATATAGCAATCAACATCATTACATGATTTTGTAACCAACCGTCTCTATAGCTCCGATA CAGTGCTAGTTTCACATCGTAATAATTAAAGAGTATAATAATAAATCGAGGTGTACTTCTCATCGA TGAAGTGATGTGCCGCTTAGCTAAATTAAACTCGTATGCGAAAAATCAGTATATGTCCGGTTAATT TCTAAGAGAGAGATTGAGAGAGAATAATTGCGCCCCTCCAAATCCCCCTCTTGGACGTTAGGGAGC TATATAGACGGTATTGCTAAGTGCGATGTGTACATAACGTACCTGTCGTAGGAACATTTCTCATCC AAATTAAGTAGTAATGCATGGCATGAAATCCATTTTTGTATTTTGCATGGCAAAGAATGACAACAA GGAATACACTAGCTAGCCCTGCCCTTTTTCAATTTAATTTAACATCAAACTTAGTTATTGTATTTC TTTTGTCAGAATAGCTGGCATTGCAAAATACA RC_9 ACGTGCTTTTCCAATTGTCATGAGGAGAGAACTTGTTAAGTTGCACACATGTGTGCTTTGCATGTC TTCAGGTTACCTGGAAGGGCATTTGCAAGGAGAGGCCATGTATGGCTCACTGGAGCAAATGAAGTT GACAGCAAAGTATTCCTAAGAGCAATTCTTGCCAAGGTTCAGCCATCACCTTCTCTTACCTATTTT TCACTCTGAATGCCAACAGTGCTTTGCACATTGTAGTCTGTTTGCAGACTGCAAATGATGACCATA ATCAGATCAGAAAATAAAATAATATTATATACTTTTTGAGCCAGCTAGCAAGAATATGTAACAATA ATTCTCCTTTTTTTTTCTTGTTCTTTTCCCTGATGTGGTGCATAACAAATAACCAAACTGATGAAT GGCAGAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTACTATGAAATTTTGTG GCTTTTGTTCATCTTTGAATGGCAGTGG RC_10 ACAAGAGGCAGAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTACTATGAAAT TTTGTGGCTTTTGTTCATCTTTGGATGCAGTGGCCATTATCTAAAAACTATGAATTTCCAGACTGC AGTTTTTATCTAATTTTGTGACTTTGTACATCAGACAGTTGTGTGCATTCCTGTTGTCGATGGCGT CCTGGAAATTGGAACTACGGAAAAGGTGATTTCGTATATTATCAGCTGACAATCTAATTATATGGG CCATATAATTAAGTATAAATCAAAATACCTCATAATATATTATAAAGTATCTAATGTGATTATGTG AATATTGGCTATTTCAATGTAATTTGATATATGAAACTGATAATCCTCTGAAACTCCGTAAGGATC AAACTAATCAAAATGTATATATTTTCAAGGTGGAGGAAGATATGGGCCTGATTCAGTATGCAAGGG GCATCTTCATGGATCAACATGGCATCCACATGAAGCCCTACACTCAAA 49 AF11 AF12 AF13 AF14 AF15 RC_11 CATCCACATGAAGCCTACCCTCTCACAGCACTCAACATCCAACCCAGTCACCCACTGTACTCATCA GCATCCAATCCAGGTTCAGATGCAACTAGGTATCACCAGCCAAACAAAGTTTGATTATTCAGATGA GCTCAATGCAGATGAGGAGAATGATGACACAGAAGAAGAGGGCATGTCAGGTTCAGACACTAACAA CACTGACACTGAAAGGAATTCAGGCCAGCTGCAACTTCAAATGCAAGACCAACTGAACATGGTGAG CAATGACCACCAGACAATACCAAATAATGCAGTTTCCAGTGAGCTAATGCAGTGTGAGATGTCAGA AGTGGTAAGAGATGGCTGCTCAAATAATATTTTAGAGGATGAAATCCAAATGCTGATGGATTGCCA AAACAGTAATTGTCAGTTAAATTTGCAAGGGCCAGATGAGCCTTGTCACTCTTGGCATTTTCTCTG CGAGGAGTTACAAAATGATTACCAGCCAGGTATTACATTTGAGAAGATAATCCTTCAAAAGCACCC TTGTTCCATCCCTTCTCAATCTGACTCATACCCATA RC_12 TCCTATCCTGGTCTCTNTGTGATTTATATGTATATTTATATTTTTTATTTATATTTTTTATATTTT TATTATTAATTTTATTATTATNTATTTATATATATATATATATATATATATNTATATATATATTTA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATGTATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATAATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATAT RC_13 AGAGAGATGTTCCAGTAGTCACTGCAGTTACAGGGGAGCAGAAACACCTGAATCAAGGGGCGGGAA AGGCGCAAGTGGATGCCATCCAAGGTGATTTCAGTGCCAACCATGTGCTGAAAGAGAGGAGAAGAA GAGAGAAGCTCAATGAGAAGTTCATAATTCTGCGATCTTTGGTACCTTTCATGACAAAGGTAATTA AGTACTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGCAAAT ATGTAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAAAATA AAACATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCACATAT TTTAAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAAGCCT CTAACGAACATTGGAAGATCCTCACTAATGGGCAGCATGA RC_14 GAACTTGTTACCAAATAAACCTACTCCGAACCCCATCCCCCTATACAAAAGCTGAGTTCTCGGCCC TGTGGTACGCATGGACCACCTCTCTAGTCGGTGTTCTCATCTGTTGCGGGCTAGCTTATTCTTTGG GACTGGTATTGTGGGGTTTGCTCCGGTCAATTTTCTCAACCCCCAATAGACCTTGGTCGGCTCCTA AATTGTATGGTTACAGTGACATGAGGCTCTCTGAGTATATTGCTTTGATTTGGAACATAATCTCTC CATCCTCCAATTTGACAA RC_15 TCCCGGCGGGGTGGAGAAACGACGGTGTCCAGATGGTTCCACCCGATGGAAAGTTGCAGATGGCGT CTTTACTAAAGGGGGGAGCCAGATAATGTATTATGTGGCTCGGTGATAGTCCTCAATTTAATTCTC CTCGCATGGCATGAAAATTCACTTTTGTGTGGGAATAGGGGCTATGCAACGTGGCTGTCAAAAAGA GACACTCATCGGTGATACTTCCTACTCCCCTGAAGATGACTATACTCCAAATTAAATAGCGCCACA ATAGCTAGATCCTTTGGGAACTCTAATCTTACAGCTGTTACCCACGATAATCCAACACTGCCGAGA ATACGTCGACTACCAAGACTATTCTGAGTCGGGGGGCGGCTCCGTATACCTGTACCTATAGTAGAC TGGCTGGATCTTAATATTTCTTTTTTCTACTTTTTCAACCTTCTTCTTTTTTCTGATAAAAAATTA AAATAGAAAACATAATATTAATTCTGTATTTAAACCACTAATTAACTAATCTGTATTAAGTAAAAT AAAATATAATAAA 50 AF16 AF17 AF18 RC_16 AAGATAGTCTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGC AAATATGTAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAA AATAAAACATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCAC ATATTTTAAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAA GCCTCTAACGAACATTGGAAGATCCTCACTAATGGCAGCATGAATCTAGGGTTCACTATTTCGGAA TGCAAAATATGTTTTACCGGGCATCCGATTTTTAAAAAATTCAGAATGAAGAAAATTGAATCTTTT TTATGGATTTGAATAAATCTTGATAAATTCGAAAAAATTTCCGAACTTTTGGCCAGAAGTGAATCC TACCCGTATCCACCGGTAATAAACCTAAATTTTTGGGAGTAATGAATTAATGTTATATATAATCCA TGAATTATATAGTTCCAAACTACTCCGTAACAAATTTTCAGGAGTAGTGAAATTAATATTATTACA ATCTCAGAAAAAAATGGCAGAAACAATTAATCTGTTTTCAATTATTAATTAATTTGTTTTTGTGTC CAGATGGACAAGGCGTCGATACTAGGCGACACGATCGAGTACGTGAAGCAGCTAAGGAACCGCATA CAAGAGCTCGAGTCGTCGTCGTCGTCGTCACGAGCAGCCGCCCGGGCGCCATCGGCGGCGGCCGCC GGGAGGCGGAGGAAGAGATCCGCCGCCGCCGCCACTGCCACGGCGGCGGAAGGGATGAGCAGCAGC AATGGCCGCAATGGCGGCGAGGCGGCGGAGGTGGTGCAGGTG RC_17 TCAATCGTCGTCACGAGCAGCCGCCCGGGCGCCATCGGCGGCGGCCGCCGGGAGGCGGAGGAAGAG ATCCGCCGCCGCCGCCACTGCCACGGCGGCGGAAGGGATGAGCAGCAGCAATGGCCGCAATGGCGG CGAGGCGGCGGAGGTGGTGCAGGTGTCCATCATCGAGAGCGACGCGCTGCTGGAGCTCCGGTGCGG TTGCGGCGGCGGCGGCGGCGGTGTGGTGCTGCTCCGGGTGATGCAGGCGATGCAGGAGCTCCAGCT GGAGGTCACCGCCGTCCAGGCCTCGTGCGCCGGTGGCGAGCTGCTCGCCGAGCTGCGCGCCAAGGT CGTCGTTATGATCCTGATCTGCATGAAAATGCAAATGCAAATGCAAATGCAGAATTAAGCTTTCAT TCTTGCTCCTCTGAATTCTGAATTTATATATTCACCCTTCTTTCGATCTGCTCGTACGTTCGTTTC GCCTAAATTATGTACAAATTAACTGAATCTTTGAACTGAAAATAACTGAATCTTTTTTGTGTGTTT TTGTGTGGGTGAATTGGTTGGCGCAGGTGAAGGGGAGGAGGAGGAGCAGCATCGCTCAGGTGAAGA GGGCCATCCATCTCGTCCTCTCCTCCTCATCGATATCACCCTGAAA RC_18 CGGTTATATATAGAAAGTTCACATATATAAATCATGATAATTACCTTTTAGAAAATTCAAACAAAG AATCGATATATATAGTTCTATCTATGCATCATTTTCCTAAATCACGATAATTACCTCAAGCTTGAT AAAATGGCATTACATTACTATTATTATTACTAGAGTTTTTTCACTTCCCTGATCTAACCAATTTGC CACGATGGTTAGATATATAGATGGTTCTAATTAATCTAATGTTAATGTACTCCACATATATGATAT ATGTACATACATATCTAGTTTTAACATTTGCAAATGAATTGCAATCAGAATATATATATGCATGCA GCACAGGTGCTTGATTGCAATTATATGATTATTACTCTCTCACATATTGATATGGTAAATTTGTAT ATATTTGTGGCATGCATTCATGCACGAAGCTAGATTAATTATTAATTAATTCAGGGTGATATCGAT GAGGAGGAGAGGACGAGATGGATGGCCCTCTTCACCTGAGCGATGCTGCTCCTCCTCCTCCCCTTC ACCTGCGCCAACCAATTCACCCACACAAAAACACACAAAAAAGATTCAGTTATTTTCAGTTCAAAG ATTCAGTTAATTTGTACATAATTTAGGCGAAACGAACGTACGAGCAGATCGAAAGAAG Appendix table Sequences of 17 fragments in Nep cam den Name Rc gene Sequence of Rc gene fragment fragment AF1 RC_1 GGGATGCTACTAGCTCCTCACTCACTCGCCTTTACTTATCGATCTCGATCATCCACGAGCTAGCTA GCAGTGCTCGCCATGGCCGGCGGCGAGGCGCATGCGGCGCTGCAGGCGGTGGCGCAGAGCCTCCGG TGGACCTACAGCCTCCTCTGGCAGCTCTGCCCCCACCAAGGGTACCTACCCTACCTACCTACGACA CGATGCACAGTGTTCATCCATGGCCGGCCATGGCGGATCGTCGTCGTTGTCGATGATCATCGAAGG AAGCTAGAGGATATGGCTCAATACTTTGATAATATATATACTGATCTCTCCGTACAACAAAAATAT AAAAATTCTAGCTAGTATCGAATGAGACATATGCTATGCTAGTACTACGAATCTAAAAAGATGTAC ATATTTTGATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGACGGATGTAATAATTC TGAATTTAGTTGTGATCGCATGGCATTTATCGCGTTGGTTCCTAGCAAGAACTTTAGCATCTTTGG TGTACGGCGAGATCGGTTATATATTGTCATGGTGGAGCG 51 AF2 AF3 AF4 AF5 AF6 AF7 RC_2 TTCGAATGAGACATATGCTATGCTAGTACTACGAATCTAAAAAGATGTACATAT TTTGATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGACGGATGT AATAATTCTGAATTTAGTTGTGATCGCATGGCATGCAGGAGCTCGCTGGTGTGG GGGGAGGGGCACTACAACGGCGCCGTCAAGACGCGGAAGTCGACGGTGATGCAG CCGCCGCCGGCGGAGGAGGAGGACGACGCCGACCACGCGGCGCGCCACCGGAGC CGGCAGCTGAGGGAGCTCTACGACTGGCTGCAGCAGGCCGGGGAGAACTCCAGC GGCGGCGTGCAGACGTCGTCGACGACGGCGAGCCGGCGGCCGGGGGCGGCTCTG TCGCCGGAGGACCTGACGGAGACGGAGTGGTTCTTCCTCATGTCGGCATCCTAC TCCTTCCCTCCCGGCATCGGGTATATAATAAAAAATATAGATATAAATATTTAA GCATGCATGCATAAATTAAACCACACTTCTTGTTACTGTTTCTTGGGCAAAA RC_3 GCGGCGTGCAGACGTTCGTCGACGACGGCGTAGCCGGCGGCCGGGGGCGGCTCTGTCGCCGGAGGA CCTGACGGAGACGGAGTGGTTCTTCCTCATGTCGGCATCCTACTCCTTCCCTCCCGGCATCGGGTA TATAATAAAAAATATAGATATAAATATTTAAGCATGCATGCATAAATTAAACCACACTTCTTGTTA CGTGTTCTTGGCAAAATGATGAACAATTACCACTAATTAATTGGAGCCAGAAACCCTAAAGATTTA CCCACCTGGTTAATTAATCGGTGTGTTGATCCACGCATGCATGCATGCAGAAAATCAAGATCAGGA TAGCTCCTTTTCTTTTGCAGGTTAATTAGCTAGATCTTCACGTATAATTAGCTAGCTAGATTTTAA AATATAATTTATTCAATTTGATTTATGATTTTTATTTTTTATTTCAAATAGATACAACTGTATACA AAATTTTATTTTGGTACATACCTCCGATCCAACTACATCAGAGGTAAAAAAAAAATTAAACCGTTG GAATTGATTAGAACAAGATCGTGCGGCCAGTCCTAAGGCTCCCCAAAA RC_4 GTTGCTGATCTTCCGTATAATTAGCTAGCTAGATTTTAAATATAATTTATTCAATTTGATTTATGA TTTTTATTTTTTATTTCAAATAGATACAACTGTATACAAAATTTTATTTTGGTACATACCTCCGAT CCAACTACATCAGAGGTAAAAAAAAAATTAAACCGTTGGAATTGATTAGAACAAGATCGTGCGGTC AAATTATATCATAACTAACTTTTCTGATTCTCTAAAGCATAGAGATGTATATATACATCGTATTAT TAGGCTCTATATTTCCTGATTAACACTAGATGCATATATAATTTTGATAGTCAAAATATACTTTTG ATAGGCTCTAAAGAAAAACTTAATAACATGTACTCCCTCCATATACTTTTGATAGTCATATTTCAT CTTGACACACAGATCAAGTATAAGTAATTCTACTTATCATCCATTTAAACACGCTACTAGTTATTC CTCATAAACAAGCGATTCATTAATATTTACATTTCTCGATGCTTGTGTAGCCAATATTGTGTGGAA GAATGGAATGTCATTAAGAGGATAGGTTGTTGGATTGAAAATATACCAAAGAA RC_5 AAGGAGTCTTAAGAGGATAGGTTGTTGGATTGAAATATGCCTATCAAAAATAAATTTTTAGATTTG AAAATATGCCTATCAAAAGTAGATGGAGGGAGTATTAATTAATGTGAATTTCCAATCCTACTGTTG TGATATTAGGCTTTGTACCTTCTTGTCCAGGAGGTATATATATGGCTCTTTTAAGGATGGGAGAAA ATATCATCTTTAATACAACTATATATGGCTTTTGTTTGATAAATACAACTTTTATTTTGTATGAAT ACAAATATATTGATAAATATCCACCATTATAATCCTAACCCATTAGGATCATATGGTGTATATTTT TTTAACTATTTGTTTTTTATAAATTAATATTAAGAGATCACAATAAAAATATAGTATTATGAAAGT ACTCTTAACAACATATCCAATGATAAAATTATTATTATTACAAAATATAGTGGTCAAATTGTATAG AATTCAATAGCCTGATTTTATGACGTCAAGTAAATTAAATAAAGAATGAAGGTAGTGCTAGAGTGA TCAAAA RC_6 ATAAAGATGAAGGTAGTGCTAGAGTGATCAACAATATCTCTCCTAAAATATGTCCTATAAGTTTTA CTCCATAAATCCAAGGGTCAAAAGTTGTTGGGTTATTTTTTTAGATAATAACATACTACCCCTTTT CAAAATGTATGATTCTATTGACTTTTTGCACAACATTTAACCATTTGTCATATTAAAAATTAGTAT AAACATCTAAAAATATAAGTTACAATTATATTTTATTTGATGATAAAACAACTCACAACAAAATAA ATAATATTTATATAATCTTTTTGGAATAAAACGAATGATCAAACATTATTCAAAAAGTCAATGGTA TAGTACGTTTTGAAATTGATAGACTATGAGAGCAAAATTTTGAGATAACATGGAAAATTATCCTCT TAGACATTGCACTGTGTAATAATTAATAATAATGAATGAAAGGCTAAGACTTTTCTTCCACCTTAT ATAAGTGGTTGAATATATAGCAATCACATCATTACATGATTTTGTAACCAACCGTCTCTATAGCTC CGAATACAGTGCAATA RC_7 AATGGTGAATATAGCAATCACATCATTACATGGATTTTGTAACCAACCGTCTCTATAGCTCCGATA CAGTGCTAGTTTCACATCGTAATAATTAAAGAGTATAATAATAAATCGAGGTGTACTTCTCATCGA TGAAGTGATGTGCCGCTTAGCTAAATTAAACTCGTATGCGAAAAATCAGTATATGTCCGGTTAATT TCTAAGAGAGAGATTGAGAGAGAATAATTGCGCCCCTCCAAATCCCCCTCTTGGACGTTAGGGAGC TATATAGACGGTATTGCTAAGTGCGATGTGTACATAACGTACCTGTCGTAGGAACATTTCTCATCC AAATTAAGTAGTAATGCATGGCATGAAATCCATTTTTGTATTTTGCATGGCAAAGAATGACAACAA GGAATACACTAGCTAGCCCTGCCCTTTTTCAATTTAATTTAACATCAAACTTAGTTATTGTATTTC TTTTGTCAGAATAGCATGGCATTGCAAAATAC 52 AF9 AF10 AF11 AF12 AF13 AF14 RC_9 GTGCTTTTCCAATTGTCATGAGGAGAGAACTTGTTAAGTTGCACACATGTGTGCTTTGCATGTCTT CAGGTTACCTGGAAGGGCATTTGCAAGGAGAGGCCATGTATGGCTCACTGGAGCAAATGAAGTTGA CAGCAAAGTATTCCTAAGAGCAATTCTTGCCAAGGTTCAGCCATCACCTTCTCTTACCTATTTTTC ACTCTGAATGCCAACAGTGCTTTGCACATTGTAGTCTGTTTGCAGACTGCAAATGATGACCATAAT CAGATCAGAAAATAAAATAATATTATATACTTTTTGAGCCAGCTAGCAAGAATATGTAACAATAAT TCTCCTTTTTTTTTCTTGTTCTTTTCCCTGATGTGGTGCATAACAAATAACCAAACTGATGAATGG CAGAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTACTATGAAATTTTGTGGC TTTTGTTCATCTTTGAAAGGCAGTGGA RC_10 CCGCAATGAGGAAGGGCGGAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTAC TATGAAATTTTGTGGCTTTTGTTCATCTTTGGATGCAGTGGCCATTATCTAAAAACTATGAATTTC CAGACAGCAGTTTTTATCTAATTTTGTGACTTTGTACATCAGACAGTTGTGGCATTCCAGTGGTCG ATGGCGTCCCGGAAATTGGAACTCCGGAGAAGGTGATTTCGTATATGATCACCTGTAGAATCTCAA TTATATGCATCCACCATATAATCCGTGTAAAAATCAAAAACCGTTACAAAGATCTATAATACTCCT CTGATGTTTATGTTGACATATAGGTCTATATTCGGCTGTGCATCTTATACATAT RC_11 AATTTGGTATCACTGGTCATCCACATGAAGCCTACCCTCTCACAGCACTCAACATCCAACCCAGTC ACCCACTGTACTCATCAGCATCCAATCCAGGTTCAGATGCAACTAGGTATCACCAGCCAAACAAAG TTTGATTATTCAGATGAGCTCAATGCAGATGAGGAGAATGATGACACAGAAGAAGAGGGCATGTCA GGTTCAGACACTAACAACACTGACACTGAAAGGAATTCAGGCCAGCTGCAACTTCAAATGCAAGAC CAACTGAACATGGTGAGCAATGACCACCAGACAATACCAAATAATGCAGTTTCCAGTGAGCTAATG CAGTGTGAGATGTCAGAAGTGGTAAGAGATGGCTGCTCAAATAATATTTTAGAGGATGAAATCCAA ATGCTGATGGATTGCCAAAACAGTAATTGTCAGTTAAATTTGCAAGGGCCAGATGAGCCTTGTCAC TCTTGGCATTTTCTCTGCGAGGAGTTACAAAATGATTACCAGCCAGGTATTACATTTGAGAAGATA ATCCTTCAAAAGCACCCTTGTTCCATCCCTTCTCATTCTGACTCATACCCATAA RC_12 AAAAAAAAAAAAAAATATATAGATAGGTGGAAGGAGAATTCTAATCTCTGTCTCTGGTGTATCTCT AGTGTCTATCTGCTACGCAATTCTAGTCTATGATTATATCTAGTAGGTCGCCGTAATTTAAGCATT GTATAATCGTGGTAATAGCTGATACCTGTATATAATTGTTATCCTCTCTCAATTCCACACATCTCA CTCACCCGGAAACATAGAGAGATATGCCTGTGGTGCCAAATGAGTGAGCTAACTCACATTGATTGC GTTGCACACACAGACCGCTATCCAGACAGGAAACCAGACATGCCAACTGCAATAATGAATCATCCA ACGCGCTATGAAAGGCTGTTTGCATATTGGTCTCTCTTCATCTTCCTCTCTCACAGACTCTCTGCA CTCGATACATTTAGCTGCAGAGAGCGATATCAACTCACTCTATAGATGTATATACTGTATCCTACT ATATCTATGTGATATCGCTAATATATCAAGATATCTATATGCCTACATATAGCTATATACCGTAGA GATAGCACGCGTTGCTGTC RC_13 GAGATGTTCCAGTAGTCACTGCAGTTACAGGGGAGCAGAAACACCTGAATCAAGGGGCGGGAAAGG CGCAAGTGGATGCCATCCAAGGTGATTTCAGTGCCAACCATGTGCTGAAAGAGAGGAGAAGAAGAG AGAAGCTCAATGAGAAGTTCATAATTCTGCGATCTTTGGTACCTTTCATGACAAAGGTAATTAAGT ACTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGCAAATATG TAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAAAATAAAA CATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCACATATTTT AAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAAGCCTCTA ACGAACATTGGAAGATCCTCACTAATGGGCAGCATGA RC_14 CTATACCCCGATTGCCAGAATAAGTTTTACCGGGCATCCGATTTTTAAAAAATTCAGAATGAATAA AATTGAATCTTTTTTTATGGATCTTGAATAAATCTTGATAAACTCGAAAAAATTTCCGAACTTTTG GCTAGAACTGAACCCTACCCGTAACCACCGGTAATAAACCTAAATTTTGGGGAGTTATGAACTAAT GTTATATATAATCCATGAATTATATAATTCCAAACTACTCCGTAACAACTTTTCAGGCGTAGTGAC ATTAATATTATCACAATCTCCGAAAAATATGGCTCATACACTAAACCTGTCTTCTCTTCCTAATTA ACTTGTTTTCGTGTCCAGATGGACAAGGCGTCGATCCTAGGCGACACGATCGAGTACGTGAAGCAG CTAAGGAACCTCATACAA 53 AF15 AF16 AF17 AF18 RC_15 TTCTTCCTCATGGCGGCGTCCTCCTCCTACTTCCCCCCCCCCCGCATGGATATAAAAAAAATCAAT GGAAATATTGATTATGCAGACGGCTGTATAAAAAAAACACTACATGTTGATGTGTCGGTCGCTGGA TGAAGAATATTATGTATTATCAGTAGCACTCAACATCACTGAATAAGATTCAGATTAATTTCTACG TGTTGATCTATTCACGTATGATGCAAAAAAAAATTATTAAAATACATCCTTTATTTTAATTTGTAA GTTAATTATATCTATGTCAAAATAATTAGATAGATAATTATATACTTCATCAATACTGTATCATAC ATCGTTATTTATATTTACTTTAAAATATATACAAATGAAAAATTATTATTAGAAAAATAATGCTCC AATTAACCAAATATTAGAGAATTGAATATTAAATTTATAATATGAAATAATTTTATTTATATCAAA ATATCATATAAAAAATAATAATAAAATTATTTATTAAAATAAATATATATAAATATTTATAATAAA TAATAAAAATAAAATAAAATTTAAAAAAAATAAAAATAAATATAATATATAAAAAATAAAATACAA TAAATATAATA RC_16 AAGATAGTCTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGC AAATATGTAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAA AATAAAACATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCAC ATATTTTAAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAA GCCTCTAACGAACATTGGAAGATCCTCACTAATGGCAGCATGAATCTAGGGTTCACTATTTCGGAA TGCAAAATATGTTTTACCGGGCATCCGATTTTTAAAAAATTCAGAATGAAGAAAATTGAATCTTTT TTATGGATTTGAATAAATCTTGATAAATTCGAAAAAATTTCCGAACTTTTGGCCAGAAGTGAATCC TACCCGTATCCACCGGTAATAAACCTAAATTTTTGGGAGTAATGAATTAATGTTATATATAATCCA TGAATTATATAGTTCCAAACTACTCCGTAACAAATTTTCAGGAGTAGTGAAATTAATATTATTACA ATCTCAGAAAAAAATGGCAGAAACAATTAATCTGTTTTCAATTATTAATTAATTTGTTTTTGTGTC CAGATGGACAAGGCGTCGATACTAGGCGACACGATCGAGTACGTGAAGCAGCTAAGGAACCGCATA CAAGAGCTCGAGTCGTCGTCGTCGTCGTCACGAGCAGCCGCCCGGGCGCCATCGGCGGCGGCCGCC GGGAGGCGGAGGAAGAGATCCGCCGCCGCCGCCACTGCCACGGCGGCGGAAGGGATGAGCAGCAGC AATGGCCGCAATGGCGGCGAGGCGGCGGAGGTGGTGCAGGTTC RC_17 GCCGTCGTCGTCGTCACGAGCAGCCGCCCGGGCGCCATCGGCGGCGGCCGCCGGGAGGCGGAGGAA GAGATCCGCCGCCGCCGCCACTGCCACGGCGGCGGAAGGGATGAGCAGCAGCAATGGCCGCAATGG CGGCGAGGCGGCGGAGGTGGTGCAGGTGTCCATCATCGAGAGCGACGCGCTGCTGGAGCTCCGGTG CGGTTGCGGCGGCGGCGGCGGCGGTGTGGTGCTGCTCCGGGTGATGCAGGCGATGCAGGAGCTCCA GCTGGAGGTCACCGCCGTCCAGGCCTCGTGCGCCGGTGGCGAGCTGCTCGCCGAGCTGCGCGCCAA GGTCGTCGTTATGATCCTGATCTGCATGAAAATGCAAATGCAAATGCAAATGCAGAATTAAGCTTT CATTCTTGCTCCTCTGAATTCTGAATTTATATATTCACCCTTCTTTCGATCTGCTCGTACGTTCGT TTCGCCTAAATTATGTACAAATTAACTGAATCTTTGAACTGAAAATAACTGAATCTTTTTTGTGTG TTTTTGTGTGGGTGAATTGGTTGGCGCAGGTGAAGGGGAGGAGGAGGAGCAGCATCGCTCAGGTGA AGAGGGCCATCCATCTCGTCCTCTCCTCCTCATCGATATCACCCCTGAAA RC_18 CTTTACGGTTATTATATAAGAAAGTTCACATATATAAATCATGATAATTACCTTTTAGAAAATTCA AACAAAGAATCGATATATATAGTTCTATCTATGCATCATTTTCCTAAATCACGATAATTACCTCAA GCTTGATAAAATGGCATTACATTACTATTATTATTACTAGAGTTTTTTCACTTCCCTGATCTAACC AATTTGCCACGATGGTTAGATATATAGATGGTTCTAATTAATCTAATGTTAATGTACTCCACATAT ATGATATATGTACATACATATCTAGTTTTAACATTTGCAAATGAATTGCAATCAGAATATATATAT GCATGCAGCACAGGTGCTTGATTGCAATTATATGATTATTACTCTCTCACATATTGATATGGTAAA TTTGTATATATTTGTGGCATGCATTCATGCACGAAGCTAGATTAATTATTAATTAATTCAGGGTGA TATCGATGAGGAGGAGAGGACGAGATGGATGGCCCTCTTCACCTGAGCGATGCTGCTCCTCCTCCT CCCCTTCACCTGCGCCAACCAATTCACCCACACAAAAACACACAAAAAAGATTCAGTTATTTTCAG TTCAAAGATTCAGTTAATTTGTACATAATTTAGGCGAAACGAACGTACGAGCAGATCGAAAAGAAG GACATGAGCAAAGAAAAAGGG 54 Appendix table Sequences of 17 fragments in Nep cam (Hoa Binh) Name Rc gene Sequence of Rc gene fragment fragment AF1 AF2 AF3 AF4 RC_1 CTAAGCTCCTCACTCACTCGCCTTTACTTATCGATCTCGATCATCCACGAGCTAGCTAGCAGTGCT CGCCATGGCCGGCGGCGAGGCGCATGCGGCGCTGCAGGCGGTGGCGCAGAGCCTCCGGTGGACCTA CAGCCTCCTCTGGCAGCTCTGCCCCCACCAAGGGTACCTACCCTACCTACCTACGACACGATGCAC AGTGTTCATCCATGGCCGGCCATGGCGGATCGTCGTCGTTGTCGATGATCATCGAAGGAAGCTAGA GGATATGGCTCAATACTTTGATAATATATATACTGATCTCTCCGTACAACAAAAATATAAAAATTC TAGCTAGTATCGAATGAGACATATGCTATGCTAGTACTACGAATCTAAAAAGATGTACATATTTTG ATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGACGGATGTAATAATTCTGAATTTA GTTGTGATCGCCATTGGCATAAA RC_2 TCAGCTGTTCGAATGAGACATATGCTATGCTAGTACTACGAATCTAAAAAGATG TACATATTTTGATTCGTATTATTAGGATATATCACGAGTTTTTATATTTTGAGA CGGATGTAATAATTCTGAATTTAGTTGTGATCGCATGGCATGCAGGAGCTCGCT GGTGTGGGGGGAGGGGCACTACAACGGCGCCGTCAAGACGCGGAAGTCGACGGT GATGCAGCCGCCGCCGGCGGAGGAGGAGGACGACGCCGACCACGCGGCGCGCCA CCGGAGCCGGCAGCTGAGGGAGCTCTACGACTGGCTGCAGCAGGCCGGGGAGAA CTCCAGCGGCGGCGTGCAGACGTCGTCGACGACGGCGAGCCGGCGGCCGGGGGC GGCTCTGTCGCCGGAGGACCTGACGGAGACGGAGTGGTTCTTCCTCATGTCGGC ATCCTACTCCTTCCCTCCCGGCATCGGGTATATAATAAAAAATATAGATATAAA TATTTAAGCATGCATGCATAAATTAAACCACACTTCTTGTTACGTGTTCTTGGC AA RC_3 CAATGTACACCTGTAAATTAAAGAAAAAAAAAAAAAAGAGGAAGGGAATTAATTTTTTTTTTTGGC CGTCCGGCCGTTCCCCCCCCACCAAACCCCCCGCACCACCCCCCACAACAACAAAATGCCAGCGAA AACCCAAACCCAAAGAGGAGAGCAAGGGCAACGCAACCGGACGCCGCCTCCGACGGAGAGCGCAGC GGCGGAGCCAAGGCCGGCCAGGGCGGACCAAGACCACGGCCAAGGAACAGCGAGCCGGCCTAGCCG ACAGGGAACGGAACTCGAGGACAGGCGTGCGGCTGAACCCTCCGTGCAACAGCAGCGGCGAGCGTC GGGCGAGGACCGCATGAAACAGGAGGCGGGGGGCGACTACGAATCTAAAGGAGATGTACATAGACG GGATGCTGATTAGTAGGATATGTCACCAGGGGATTATCTGCGTGGCACGGAGGTAATAATAACATG AATGTAGTTGTGACGACATGGGGCA RC_4 GGGTTGCTGATCTTCCGTATAATTAGCTAGCTAGATTTTAAATATAATTTATTCAATTTGATTTAT GATTTTTATTTTTTATTTCAAATAGATACAACTGTATACAAAATTTTATTTTGGTACATACCTCCG ATCCAACTACATCAGAGGTAAAAAAAAAATTAAACCGTTGGAATTGATTAGAACAAGATCGTGCGG TCAAATTATATCATAACTAACTTTTCTGATTCTCTAAAGCATAGAGATGTATATATACATCGTATT ATTAGGCTCTATATTTCCTGATTAACACTAGATGCATATATAATTTTGATAGTCAAAATATACTTT TGATAGGCTCTAAAGAAAAACTTAATAACATGTACTCCCTCCATATACTTTTGATAGTCATATTTC ATCTTGACCCACAGATCAGGTATAAGTAATTCTACTTATCATCCATTTAAACACGCTACTAGTTAT TCCTCATAAACAAGCGATTCATTAATATTTACATTTCTCGATGCTTGTGTAGCCAATATTGTGTGG AAGAATGGAATGTCATTAAGAGGATAGGTTGTTGGATTGAAATATACCAAGG 55 AF5 AF6 AF7 AF9 AF10 AF11 RC_5 AAGGATGTAATTAAGAGGATAGGTTGTTGGATTGAAATATGCCTATCAAAAATAAATTTTTAGATT TGAAAATATGCCTATCAAAAGTAGATGGAGGGAGTATTAATTAATGTGAATTTCCAATCCTACTGT TGTGATATTAGGCTTTGTACCTTCTTGTCCAGGAGGTATATATATGGCTCTTTTAAGGATGGGAGA AAATATCATCTTTAATACAACTATATATGGCTTTTGTTTGATAAATACAACTTTTATTTTGTATGA ATACAAATATATTGATAAATATCCACCATTATAATCCTAACCCATTAGGATCATATGGTGTATATT TTTTTAACTATTTGTTTTTTATAAATTAATATTAAGAGATCACAATAAAAATATAGTATTATGAAA GTACTCTTAACAACATATCCAATGATAAAATTATTATTATTACAAAATATAGTGGTCAAATTGTAT AGAATTCAATAGCCTGATTTTATGACGTCAAGTAAATTAAATAAAGAATGAAGGTAGTGCTAGAGT GATCAAAA RC_6 TTAATAATGAGGTAGTGCTAGAGTGATCAAACAATATCTCTCCTAAAATATGTCCTATAAGTTTTA CTCCATAAATCCAAGGGTCAAAAGTTGTTGGGTTATTTTTTTAGATAATAACATACTACCCCTTTT CAAAATGTATGATTCTATTGACTTTTTGCACAACATTTAACCATTTGTCATATTAAAAATTATTAT TTTCTTTTTTTTAGACAAATTTTATTTGATGACAAAACAACTCACACCACAATACATATAATTTAA ATTATCTATATATAAAAATATATTACAAACATATATATATATATATATTATATATATATACATATA TATATATATATACAGATATATATATATATATATTCTATATAATATCATATATATATCATATATATA CTATATGATATATATATTATATACTATATGATATAGTATATTATATATACTATCCTATACTATATA TACTATATAGTACTATATAATATACTATACTATCATATCTATAATACTATAGTATACTATACTATA TTATAATATATCATACCATATACTATACTATTATATATTATAATATATATAATATATAATATATAT AAATAATATATATATATACTATATATAATAATATATATATCATATATAATACCATACTATATAATA TAATATAATATAATCATATATATAATATATAATAATATAATACTATAATATATATTATTATATACT ATAAATATATAATATATAATACTATAATACTATAATAATATAATATAATAATAAAATAATATATAA AAATATAATTATATATAAAATATAATATAATATAATAATCATAATATAACTATAATATATTAC RC_7 AAGGGTGAATATAGCAATCACATCATTACATGATTTTGTAACCAACCGTCTCTATAGCTCCGATAC AGTGCTAGTTTCACATCGTAATAATTAAAGAGTATAATAATAAATCGAGGTGTACTTCTCATCGAT GAAGTGATGTGCCGCTTAGCTAAATTAAACTCGTATGCGAAAAATCAGTATATGTCCGGTTAATTT CTAAGAGAGAGATTGAGAGAGAATAATTGCGCCCCTCCAAATCCCCCTCTTGGACGTTAGGGAGCT ATATAGACGGTATTGCTAAGTGCGATGTGTACATAACGTACCTGTCGTAGGAACATTTCTCATCCA AATTAAGTAGTAATGCATGGCATGAAATCCATTTTTGTATTTTGCATGGCAAAGAATGACAACAAG GAATACACTAGCTAGCCCTGCCCTTTTTCAATTTAATTTAACATCAAACTTAGTTATTGTATTTCT TTTGTCAGAATAGCATGCATTGCAAAAT RC_9 GCTTTTCCATTGTCATGAGGAGAGAACTTGTTAAGTTGCACACATGTGTGCTTTGCATGTCTTCAG GTTACCTGGAAGGGCATTTGCAAGGAGAGGCCATGTATGGCTCACTGGAGCAAATGAAGTTGACAG CAAAGTATTCCTAAGAGCAATTCTTGCCAAGGTTCAGCCATCACCTTCTCTTACCTATTTTTCACT CTGAATGCCAACTGAGCTTTAGCACATGTAAGAACAGTGCACAATAATGCAAAATGACTCATCATT ATTACAGAGTCTCAAAATTAAACAATATAATAATTACACTATTTTTACAACAATCTTAAGCAATGA AACAATTCATACAATGTTATATTCCATACCTTTCATTTTATTTTCTTGATTTCA RC_10 AAACGGAGAAGGCCAGAAGTGCTGGTATCCAGGTATTTGCCTCTAAAAGTAGCTACACGTTTACTA TGAAATTTTGTGGCTTTTGTTCATCTTTGGATGCAGTGGCCATTATCTAAAAACTATGAATTTCCA GACTGCAGTTTTTATCTAATTTTGTGACTTTGTACATCAGACAGTTGTGTGCATTCCTGTTGTCGA TGGCGTCCTGGAAATTGGAACTACGGAAAAGGTGATTTCGTATATTATCAGCTGACAATCTAATTA TATGGGCCATATAATTAAGTATAAATCAAAATACCTCATAATATATTATAAAGTATCTAATGTGAT TATGTGAATATTGGCTATTTCAATGTAATTTGATATATGAAACTGATAATCCTCTGAAACTCCGTA AGGATCAAACTAATCAAAATGTATATATTTTCAAGGTGGAGGAAGATATGGGCCTGATTCAGTATG CAAGGGGCATCTTCATGGATCAACATGGCATCCACATGAAGCCCTACCCTCAA RC_11 GGTTAAACTGGCAATCACATGAAGCCTACCCTCTCACAGCACTCAACATCCAACCCAGTCACCCAC TGTACTCATCAGCATCCAATCCAGGTTCAGATGCAACTAGGTATCACCAGCCAAACAAAGTTTGAT TATTCAGATGAGCTCAATGCAGATGAGGAGAATGATGACACAGAAGAAGAGGGCATGTCAGGTTCA GACACTAACAACACTGACACTGAAAGGAATTCAGGCCAGCTGCAACTTCAAATGCAAGACCAACTG AACATGGTGAGCAATGACCACCAGACAATACCAAATAATGCAGTTTCCAGTGAGCTAATGCAGTGT GAGATGTCAGAAGTGGTAAGAGATGGCTGCTCAAATAATATTTTAGAGGATGAAATCCAAATGCTG ATGGATTGCCAAAACAGTAATTGTCAGTTAAATTTGCAAGGGCCAGATGAGCCTTGTCACTCTTGG CATTTTCTCTGCGAGGAGTTACAAAATGATTACCAGCCAGGTATTACATTTGAGAAGATAATCCTT CAAAAGCCCCCTTGTTCCATCCATAC 56 AF12 AF13 AF14 AF15 RC_12 AAAAAAAAAAAATATATATACTATCTGTATAATGTAATTCCTAATATGTATCCTATTTAATATTTA TTTTATATATATTTTTTTTATATTTATTTTTATATATTTTTTATTCATTTATATTTATATATTTAT ATATATATATTTTTATATTTATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATATATATATATATATATATATAATATATATATATATATATATA TATATAATATATATATATATATAATATATATATATATATATATATATATATATATATATATATATA TAATATATATAATATATAATATATATATATATATATATATATATATATATATATATATATATATAT ATATATATATATATATATATATATA RC_13 CGAATGTTCCAGTAGTCACTGCAGTTACAGGGGAGCAGAAACACCTGAATCAAGGGGCGGGAAAGG CGCAAGTGGATGCCATCCAAGGTGATTTCAGTGCCAACCATGTGCTGAAAGAGAGGAGAAGAAGAG AGAAGCTCAATGAGAAGTTCATAATTCTGCGATCTTTGGTACCTTTCATGACAAAGGTAATTAAGT ACTCCCTCTATTTCTATAAAGCCGTATTTGACTAGTTATCTTATTTAGAAAGTATGTGCAAATATG TAAAATATAAGTCATACTTAAAGAACTTTTAATGTTATTAAATAATAAGTCACACCAAAAATAAAA CATATATATTTTTAATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCACATATTTT AAAATAGAGGGGTATTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAAGCCTCTA ACGAACATTGGAAGATCCTCACTAATGGCAGCATGAAG RC_14 CAGGGCATGAATCTAGGGTTCACTATTTCGGAATGCAAATATGTTTTACCGGGCATCCGATTTTTA AAAAATTCAGAATGAAGAAAATTGAATCTTTTTTATGGATTTGAATAAATCTTGATAAATTCGAAA AAATTTCCGAACTTTTGGCCAGAAGTGAATCCTACCCGTATCCACCGGTAATAAACCTAAATTTTT GGGAGTAATGAATTAATGTTATATATAATCCATGAATTATATAGTTCCAAACTACTCCGTAACAAA TTTTCAGGAGTAGTGAAATTAATATTATTACAATCTCAGAAAAAAATGGCAGAAACAATTAATCTG TTTTCAATTATTAATTAATTTGTTTTTGTGTCCAGATGGACAAGGCGTCGATACTAGGCGACACGA TCGAGTACGTGAAGCAGCTAAGGAACCGCATACAAGAGCTCGAGTCGTCGTCGTCGTCGTCACGAG CAGCCGCCCGGGCGCCATCGGCGGCGGCCGCCGGGAGGCGGAGGAAGAGATCCGCCGCCGCCGCCA CTGCCACGGCGGCGGAAGGGATGAGCAGCAGCAATGGCCGCAATGGCGGCGAGGCGGCGGAGGTGG TGCAGGTGTCCATCATCGAGAGCGACGCGCTGCTGGAGCTCCGGTGCGGTTGCGGCGGCGGCGGCG GCGGTGTGGTGCTGCTCCGGGTGATGCAGGCGATGCAGGAGCTCCAGCTGGAGGTCACCGCCGTCC AGGCCTCGTGCGCCGGTGGCGAGCTGCTCGCCGAGCTGCGCGCCAAGGTCGTCGTTATGATCCTGA TCTGCATGAAAATGCAAATGCAAATGCAGATGCAGAATTAAGCTTTCATTCTTGCTCCTATGAA RC_15 CGTCGACGACGAACGGAGAAGGCGGCGGCGGGGGGCTGCTCACGGAGAGCACCCGGCGGCGTCTGT AGTGGTTCATCATGATCGTCAGCATCCTACTCCTCCCCTCCCGGCATGGGGAATATAATAAAAAAT GTAGATAAAAATTATTAATCCTGCCTGCATAAATTAAACCACACTTGTTGTTACTTGTTCTTGGCA AAATGATGAACAATTACCACTAATTAATTGGAGCCAGAAACCCTAAAGATTTACCCACCTGGATTA ATTAAGCGTGTGTGTTGATCCACGCATGCATGCATGCAAAAAAATCGGATGCTAGATAGCTCCTTT TCTTTTGCAGGTTAATTAGCTATATCTTCACGTATAATTAGCTAGCTAGATATTAAAATATAATTT ATTCAATTTGATTTATGATTTTTTATTTTTATATTCAATATAGAATACGAACTGGTATAACTAAAT TTTTATGTTCTGGTAACATTACCATCCGAAATCCACTCACATCGAGG 57 AF16 RC_16 CCAGCAGCATTAGACTAGTTATACTCTATTTAGAACAGTATGTGCAAATATGTAAAATATAAGTCA TACTTAAAGAACTTTTAATGTTATGAAATAATAAGTCACACCAAAAATAAAACATATATATTTTTA ATAAGATAAATGATTAAATGTATATATAAAAATTAATAGCGTCACATATTTTAAAATAGAGGGGTA TTTAAGTACCCACAGGATCATCAAAATTCAGTTATCTTTTCTTAAGCCTCTAACGAACATTGGAAG ATCCTCACTAATGGCAGCATGAATCTAGGGTTCACTATTTCGGAATGCAAAATATGTTTTACCGGG CATCCGATTTTTAAAAAATTCAGAATGAAGAAAATTGAATCTTTTTTATGGATTTGAATAAATCTT GATAAATTCGAAAAAATTTCCGAACTTTTGGCCAGAAGTGAATCCTACCCGTATCCACCGGTAATA AACCTAAATTTTTGGGAGTAATGAATTAATGTTATATATAATCCATGAATTATATAGTTCCAAACT ACTCCGTAACAAATTTTCAGGAGTAGTGAAATTAATATTATTACAATCTCAGAAAAAAATGGCAGA AACAATTAATCTGTTTTCAATTATTAATTAATTTGTTTTTGTGTCCAGATGGACAAGGCGTCGATA CTAGGCGACACGATCGAGTACGTGAAGCAGCTAAGGAACCGCATACAAGAGCTCGAGTCGTCGTCG TCGTCGTCACGAGCAGCCGCCCGGGCGCCATCGGCGGCGGCCGCCGGGAGGCGGAGGAAGAGATCC GCCGCCGCCGCCACTGCCACGGCGGCGGAAGGGATGAGCAGCAGCAATGGCCGCAATGGCGGCGAG GCGGCGGAGGTGGTGCAGGTGTCCATCAATCGAGAGCAAA AF17 RC_17 CGGGGGGGGGGGCCGGAACCCGCTACACGTACGCTTATCAACAACGGATCTCTAGAAGGCGAGAGA GGGCGATGTATCCATATCGAATACATCTAGAGTATATTTATAGATGGGGGCAGGTGAGCTTCTCCA CTCAAATAGGGTGGGCGCGCATGGAATTTAAACACTTCTGTGCAGAATACGGGATATGT RC_18 TTTCGGTTTTTATAGAAAGTTCACATATATAAATCATGATAATTACCTTTTAGAAAATTCAAACAA AGAATCGATATATATAGTTCTATCTATGCATCATTTTCCTAAATCACGATAATTACCTCAAGCTTG ATAAAATGGCATTACATTACTATTATTATTACTAGAGTTTTTTCACTTCCCTGATCTAACCAATTT GCCACGATGGTTAGATATATAGATGGTTCTAATTAATCTAATGTTAATGTACTCCACATATATGAT ATATGTACATACATATCTAGTTTTAACATTTGCAAATGAATTGCAATCAGAATATATATATGCATG CAGCACAGGTGCTTGATTGCAATTATATGATTATTACTCTCTCACATATTGATATGGTAAATTTGT ATATATTTGTGGCATGCATTCATGCACGAAGCTAGATTAATTATTAATTAATTCAGGGTGATATCG ATGAGGAGGAGAGGACGAGATGGATGGCCCTCTTCACCTGAGCGATGCTGCTCCTCCTCCTCCCCT TCACCTGCGCCAACCAATTCACCCACACAAAAACACACAAAAAAGATTCAGTTATTTTCAGTTCAA AGATTCAGTTAATTTGTACATAATTTAGGCGAAACGAACGTACGAGCAGATCGAAAGAAGGCCATG AGCAAAGAAAAAG AF18 58