3santos-coffee varieties.pmd D.M. C. Santos et al. 5 SCIENCE DILIMAN (JANUARY-JUNE 2016) 28:1, 5-16 Simple Sequence Repeat Analysis of Selected NSIC-registered Coffee Varieties in the Phil ippines Daisy May C. Santos* University of the Philippines Diliman Carla Francesca F. Besa University of the Philippines Diliman Angelo Joshua A. V ictoria University of the Philippines Diliman Ernelea P. Cao University of the Philippines Diliman ABSTRACT C o f f e e (Coffea s p . ) i s a n i m p o r t a n t c o m m e r c i a l c r o p w o r l d w i d e . T h r e e species of coffee are used as beverage, namely Coffea arabica, C. canephora, and C. liberica. Coffea arabica L. is the most cultivated among the three coffee species due to its taste quality, rich aroma, and low caffeine content. Despite its inferior taste and aroma, C. canephora Pierre ex A . Froehner, w h i c h h a s t h e h i g h e s t c a f f e i n e c o n t e n t , i s t h e s e c o n d m o s t w i d e l y cultivated because of its resistance to coffee diseases. On the other hand, C. l iberica W.Bull ex Hierncomes is characterized by its very strong taste and flavor. The Philippines used to be a leading expor ter of coffee until coffee rust destroyed the farms in Batangas, home of the famous Kapeng Barako. The country has been attempting to revive the coffee industry by focusing on the production of specialty coffee with registered varieties on the National Seed Industry Council (NSIC). Correct identif ication and i s o l a t i o n o f p u r e c o f f e e b e a n s a r e t h e m a i n f a c t o r s t h a t d e t e r m i n e cof fee’s m a r ke t v a l u e . Lo c a l f a r m s u s u a l l y m i s i d e n t i f y a n d m i x cof fee beans of different varieties, leading to the depreciation of their value. This study used simple sequence repeat (SSR) markers to evaluate and distinguish Philippine NSIC-registered coffee species and varieties. The _______________ *Corresponding Author ISSN 0115-7809 Print / ISSN 2012-0818 Online S i m p l e S eq u e n ce Re p e a t A n a l y s i s of S e l ec ted N S I C- r eg i s tered Coffee 6 n e i g h b o r - j o i n i n g t r e e g e n e r a t e d u s i n g PA U P s h o w ed h i g h bootstrap support, separating C. arabica, C. canephora, and C. liberica from each other. Among the twenty primer pairs used, seven were able to distinguish C. arabica, nine for C. liberica, and one for C. canephora. Keyword s: Coffea, NSIC-registered varieties, SSR INTRODUCTION Coffee is an economically important crop in the global market. It belongs to the caffeine-containing subgenus Coffea from the family Rubiaceae, which comprises over a hundred species originating from the African region (Charrier and Berthaud 1985). Among the coffee species used for commercial consumption, C. arabica L. is the most cultivated, accounting for 70% of the global coffee production. It is the only allotetraploid in the genus and is self-pollinating. This species also has the highest market value (Tornincasa et al. 2010) because of its low caffeine content, excellent taste, and aroma (Vidal et al. 2010; Vieira et al. 2010). The species C. canephora Pierre ex A. Froehner is second to C. arabica in terms of production, contributing the remaining 30% of global coffee production. It has certain advantages in terms of production due its high-yielding properties and tolerance to diseases. However, its taste, which is characterized as woody bitter and of high caffeine content, is inferior to C. arabica (Reyes 2010). Most studies report that only C. arabica and C. canephora are cultivated for commercial consumption. The Philippines is one of the few countries that commercially produce, in addition to C. arabica and C. canephora, varieties of the species C. liberica W. Bull ex Hierncomes. The Liberica variety, C. liberica var. Liberica, was an economically important commodity during the 1930s. Locally known as the Kapeng Barako, it is distinguished for its strong, woody, and bitter taste, acidic aftertaste, and pungent aroma. Apart from its strong taste, this variety also possesses desirable reproductive characteristics in terms of fruit clusters, bean size (the largest among the four varieties), and low caffeine content (N’Diaye et al. 2005). Coffea liberica var. Dewevrei, commonly known as Excelsa coffee, has a woody taste, and sweet, fruity aroma (Reyes 2010). The identity and purity of the coffee produce determine its market value. Owing to the economic importance of coffee, it is of interest to assess its genetic diversity D.M. C. Santos et al. 7 and to come up with markers that will identify and distinguish species, as well as varieties within a species. Since morphological methods are sometimes not reliable in differentiating coffee species and varieties, molecular techniques are being used and developed to address this concern. The CBOL (Consortium for the Barcoding of Life) Plant Working Group has recommended two universal plant barcodes for species identif ication, namely the matK and rbcL genes (Janzen 2009). These two genes have been used in verifying the identities of the coffee species in the farms located in Cavite, Philippines. The said genes were able to distinguish among the species C. arabica, C. canephora, and C. liberica. However, the varieties C. liberica var. Liberica and C. liberica var. Dewevrei were not successfully differentiated and clustered together in a single clade (Cao et al. 2014). The matK and rbcL markers could discriminate between species but not varieties within species. Microsatellite or simple sequence repeat (SSR) markers are short, tandem repeats present in the coding and non-coding portions of the genome (Wang et al. 2009). SSRs require only a small amount of DNA for polymerase chain reaction (PCR)-based screening and can reveal multiple alleles at a single locus. Automated allele detection and sizing are also readily available (Schlotterer et al. 2000). The abundance and highly polymorphic property of SSRs make it a good marker for plant genetic studies, identif ication of cultivars, and evaluation of varieties with a narrow genetic base (Vieira et al. 2010; Wang et al. 2009). SSRs have been used in varietal identif ication and the evaluation of genetic diversity in C. arabica varieties (Vieira et al. 2010). In 2012, low genetic diversity was observed in the C. arabica populations in the Nicaraguan regions due to their narrow genetic base, but signif icant differentiation was found among the varieties (Geleta et al. 2012). Both C. arabica and C. canephora have also been shown to have narrow diversity using SSR markers (Anthony et al. 2001; Anthony et al. 2002; Lashermes et al. 1999). In other studies, C. arabica DNA f ingerprinting using SSR markers has also been developed as a method to test against C. canephora, in order to ensure the authenticity of the coffee products sold in the market (Tornincasa et al. 2010). SSRs have also been used to evaluate leaf miner resistance in Arabica coffee (Pereira et al. 2011). The diversity of the C. canephora gene pool was also assessed using SSRs (Prakash et al. 2005). Since coffee variety misidentif ication and coffee bean sample impurity are major factors that affect the income of small-scale farmers, this study aims to identify potential molecular markers with different SSR primers for variety identif ication S i m p l e S eq u e n ce Re p e a t A n a l y s i s of S e l ec ted N S I C- r eg i s tered Coffee 8 using NSIC-registered varieties as standards. The NSIC under the Department of Agriculture, Bureau of Plant Industry was established in 1992 under Republic Act 7308. This off ice functions to approve and register crop varieties. Currently, there are 22 registered coffee varieties across the country (NSIC 2012). MATERIALS AND METHODS Plant Material and DNA Extraction NSIC-registered coffee samples were collected from Benguet, Cavite, and Bukidnon (Table 1). Two plants from each available variety were collected. Around 100 mg of young leaves were obtained from each plant for DNA extraction. Genomic DNA was extracted using Qiagen DNeasy Plant Mini Kit (Qiagen, Valencia, CA, USA) according to the manufacturer’s instructions. Automated quantification of the amount and purity of the extracted DNA was performed using Nanodrop. On average, about 100 ng per μL of DNA was extracted for each specimen. Polymerase Chain Reaction and Electrophoresis Twenty SSR primers reported in published literature (Table 2) were used for amplification in each specimen. The concentration of the PCR components for a 14 μL reaction were as follows: 3.44 μL Qiagen master mix, 1.2 μL Q buffer, 0.5 μL 25 mM MgCl2, 0.24 μL 10 μM primers, 7.38 μL DNAse/RNase-free water, and 1.0 μL 20 ng DNA. The following PCR conditions were used: initial denaturation at 94ºC for 10 min; 35 cycles of denaturation at 94ºC for 30 s, annealing at 50ºC for 30 s, and extension Species Identity Variety Source Coffea arabica Red Bourbon Bureau of Plant Industry, Baguio City, Benguet Coffea arabica Yellow Bourbon Bureau of Plant Industry, Baguio City, Benguet Coffea arabica Caturra Bureau of Plant Industry, Baguio City, Benguet Coffea canephora Ivory Coast 2 Cavite State University, Indang, Cavite Coffea canephora Ivory Coast 7 Cavite State University, Indang, Cavite Coffea canephora Ivory Coast 8 Cavite State University, Indang, Cavite Coffea canephora S247 Cavite State University, Indang, Cavite Coffea liberica BS1 (for registry) Cavite State University, Indang, Cavite Coffea canephora FRT23 Nestle Philippines, Inc., Malaybalay, Bukidnon Coffea canephora FRT 65 Nestle Philippines, Inc., Malaybalay, Bukidnon Table 1. NSIC varieties used in this study D.M. C. Santos et al. 9 at 72ºC for 1 min; and f inal extension at 72ºC for 7 min (Teressa et al. 2010). The PCR products were run in 2% agarose gels for conf irmation. For better resolution of the bands, the PCR products were run in 10% native polyacrylamide gels. Both 100 bp (KAPA) and 25 bp (Bioline) DNA ladders were used as molecular weight markers. Primer Name Sequence Repeats Reference ssrR209 F 5’CGGGGGTAAAAAGATTGTAA3’ GA (16) Teressa et al. 2010 ssrR209 R 5’TTGGTGGGAGGGGAGTA3’ ssrR268 F 5’GTATCCCACAATGAAATCAC3’ GA (19) Teressa et al. 2010 ssrR268 R 5’AGTAGAATTTTCAACATATAAG3’ SSR124577 F 5’GATGGCTTTTCTCCGTTATCC3’ AAG (6) Teressa et al. 2010 SSR124577 R 5’GGATTCGACTGCTGGATGAT3’ SSR122850 F 5’TCCAGTTTGATCAGCAACCA3’ (AGAG)3 Teressa et al. 2010 SSR122850 R 5’CCATCTTGGGGATAGAGCAA3’ SSR124195 F 5’ATCCCCATCAGAAGACCTCA3’ (AGC)6 Teressa et al. 2010 SSR124195 R 5’CCTCCACCGCCTGTTTATTA3’ SSR123557 F 5’ATCTCCTCGTTCTTCCCCAT3’ CTCT (4) Teressa et al. 2010 SSR123557 R 5’GCTTGTAGCAGGCAGGAAAC3’ ssrCMA008 F 5’CATTCTGGTCCTGATGCTCT3’ (CT)14. .(TG)10 Teressa et al. 2010 ssrCMA008 R 5’TCATTCACTTATTAACGTCCATC3’ M-24 F 5’GGCTCGAGATATCTGTTTAG3’ Not specified Bigirimana et al. 2013 M-24 R 5’TTTAATGGGCATAGGGTCC3’ Sat235 F 5’TCGTTCTGTCATTAAATCGTCAA3’ Not specified Bigirimana et al. 2013 Sat235 R 5’GCAAAATCATGAAAATAGTTGGTG3’ Sat172 F 5’ACGCAGGTGGTAGAAGAATG3’ Not specified Bigirimana et al. 2013 Sat172 R 5’TCAAAGCAGTAGTAGCGGATG3’ Sat227 F 5’TGCTTGGTATCCTCACATTCA3’ Not specified Bigirimana et al. 2013 Sat227 R 5’ATCCAATGGAGTGTGTTGCT3’ Sat229 F 5’TTCTAAGTTGTTAAACGAGACGCTTA3’ Not specified Bigirimana et al. 2013 Sat229 R 5’TTCCTCCATGCCCATATTG3’ Sat254 F 5’ATGTTCTTCGCTTCGCTAAC3’ Not specified Bigirimana et al. 2013 Sat254 R 5’AAGTGTGGGAGTGTCTGCAT3’ ssrCMA059 F 5’GATGGACAGGAGTTGATGGT3’ (CT9)(CA)8 Teressa et al. 2010 ssrCMA059 R 5’TTTTAACACTCATTTTGCCAAT3’ ssrCMA198 F 5’AGCAACTCCAGTCCTCAGGT3’ (TG)9(AG)18 Teressa et al. 2010 ssrCMA198 R 5’TGGAAGCCCGCATATAGTTT3’ SSRCa068 F 5’ATGTTGTTGGAGGCATTTTC3’ (AGG)7//(GAA)4 Missio et al. 2011 SSRCa068 R 5’AGGAGCAGTTGTTGTTTTCC3’ SSRCa087 F 5’TCACTCTCGCAGACACACTAC3’ (TC)22 Missio et al. 2011 SSRCa087 R 5’GCAGAGATGATCACAAGTCC3’ SSRCa094 F 5’GTGTCCTAGGGAAGGGTAAG3’ (TC)4(TTCT)3// Missio et al. 2011 (TTTCCT)3 (TTCT)5 SSRCa094 R 5’GAGTGCTAGGAGAGGGAGAG3’ SSRCa091 F 5’CGTCTCGTATCACGCTCTC3’ (GT)8(GA)10 Missio et al. 2011 SSRCa091 R 5’TGTTCCTCGTTCCTCTCTCT3’ Sat207 F 5’AAGCCGTTTCAAGCC3’ Pereira et al. 2011 Sat207 R 5’CAATCTCTTTCCGATGCTCT3’ Table 2. Primer sequences used for SSR analysis S i m p l e S eq u e n ce Re p e a t A n a l y s i s of S e l ec ted N S I C- r eg i s tered Coffee 10 Data Analysis The PCR products were evaluated by scoring the presence (1) or absence (0) of clear and unambiguous bands. A neighbor-joining tree with 1,000 bootstrap replicates was constructed using PAUP version 4.0b10 for Microsoft Windows 95/ NT and viewed using TreeExplorer 2.12 by Koichiro Tamura 1997-1999. Pairwise genetic distances were also calculated using PAUP. RESULTS AND DISCUSSION A total of 236 unique bands were identif ied from the 20 SSR markers. Based on the neighbor-joining tree generated, the C. arabica, C. canephora, and C. liberica species were differentiated into separate clades (Figure 1). Of the 20 SSR markers, seven primer pairs distinguished C. arabica, nine for C. liberica, and one for C. canephora (Table 3). This shows that the SSR markers can be used in delineating species despite Figure 1. Neighbor-joining tree of 20 NSIC-registered coffee varieties generated from banding prof iles from 20 microsatellite markers. Branch lengths are drawn to scale and represent uncorrected p-distances. Bootstrap supports of 1000 replicates are shown. D.M. C. Santos et al. 11 Pairwise comparisons p Average between species (n=108) 0.382 Average between varieties of the same species (n=72) 0.285 Average between varieties of C. arabica (n=12) 0.060 Average between varieties of C. canephora (n=72) 0.330 Average within varieties (n=10) 0.034 Red bourbon (n=1) 0.017 Yellow bourbon (n=1) 0.008 Caturra (n=1) 0.009 Yellow Bourbon and Caturra combined (n=4) 0.024 Ivory Coast2 (n=1) 0.083 Ivory Coast7 (n=1) 0.068 Ivory Coast8 (n=1) 0.072 S247 (n=1) 0.000 FRT23 (n=1) 0.021 FRT65 (n=1) 0.004 BS1 (n=1) 0.057 Table 4. Generic d istances in coffee species and varieties. n, number of pairwise comparison; p, uncorrected d istance Table 3. Species d istinguished by each primer pair Primer pair Diagnosable species ssrR209 - ssrR268 - SSR124577 C. arabica SSR122850 C. liberica SSR124195 C. arabica SSR123557 C. arabica ssrCMA008 C. arabica, C. liberica, C. canephora M-24 C. liberica Sat235 - Sat172 - Sat227 C. liberica Sat229 C. liberica Sat254 C. liberica ssrCMA059 C. arabica, C. liberica ssrCMA198 C. arabica SSRCa068 C. arabica SSRCa087 - SSRCa094 - SSRCa091 C. liberica Sat207 C. liberica S i m p l e S eq u e n ce Re p e a t A n a l y s i s of S e l ec ted N S I C- r eg i s tered Coffee 12 very high polymorphisms. In par ticular, the ssrCMA008 primer pair was able to differentiate the three species. Teressa et al. (2010) used this primer pair to compare varieties of C. arabica. This is the f irst study that demonstrates its utility for species diagnosis. A 100% bootstrap support was observed for C. arabica and C. liberica species, whereas the support for C. canephora was only at 50%. The low bootstrap support for C. canephora is likely due to the large genetic distance between the Ivory Coast and S247 varieties from Cavite, and the FRT varieties from Bukidnon. The average genetic distance among varieties of C. canephora (p = 0.330) was comparable to the distances among species (p = 0.382; Table 4). The SSR markers were also able to differentiate among the varieties. Bootstrap supports of 100% were observed for the Red bourbon, Ivory Coast 2, Ivory Coast 7, S247, FRT23, FRT65, and BS1 varieties. A bootstrap support of 97% was observed for Ivory Coast 8. Bootstrap supports of 95% and 88% were observed for Yellow Bourbon and Caturra varieties, respectively. Among the C. arabica varieties, the red bourbon variety can be distinguished from the others using the SSR124577 (Figure 2) primer pair. The allele number for this primer pair was higher in this study (n=8) compared to that of Teressa et al. (2010), indicating higher diversity among the C. arabica varieties in the Philippines. The Red bourbon variety was shown to be distinct: a 150-bp band from SSR124577, and 150-bp and 350-bp bands from SAT229 primer pairs can distinguish the Red Bourbon from the other C. arabica varieties. The Yellow bourbon and Caturra varieties clustered together with 93% bootstrap support. Although the bootstrap support for each of these clades is moderately high, the values obtained were lower compared to the support for the clades of the other varieties (Figure 1). The average pairwise Figure 2. Banding patterns observed for the different Coffea varieties using the SSR124577 marker. D.M. C. Santos et al. 13 genetic distance within these two varieties combined is small (p = 0.024) and is even lower than the average genetic distance within single varieties (p = 0.033; Table 4). These two varieties were distinguished by the SSRCa087 primer pair. Apart from this, they share the same banding prof ile based on the other markers. Moreover, the 140-bp, 1,000-bp, and 1,200-bp bands from SSR124577 (Figure 2) were found to be unique to Yellow bourbon and Caturra. These varieties are commonly considered to be identical, but were registered as distinct varieties (Prof. Valentino Macanes, pers. comm.). Results in this study show partial support for this claim, but the current dataset is insuff icient to generate conclusions, considering that the two varieties did form distinct clades. The Yellow bourbon and Caturra varieties were observed to have leaves that are similar in shape and size, but Caturra had shorter internodes. According to the NSIC registry (NSIC 2012), they also differ in berry color, but this was not observed in this study because there were no berries during the time of sampling. The SAT235 is linked to disease resistance against coffee berry disease (Gichimu et al. 2014; Gichuru et al. 2008). It is not clear from these papers, however, what fragment size correspond to the marker for the disease. Based on the NSIC registry (NSIC 2012), only IC8 has a record of moderate resistance against coffee berry disease. The bands exhibited by IC8 for the SAT235 primer pair is shared by other varieties of C. canephora, except for the FRT varieties from Bukidnon. No entries are available for other varieties. Among the C. canephora varieties, the FRT varieties developed by Nestle Philippines, Inc. in Bukidnon highly diverged from the Ivory Coast and S247 varieties from Cavite—a phenomenon manifesting even in terms of morphology. FRT varieties take a much longer time to flower and fruit, but produce greater yield and more berries per leaf node. These results show the potential of SSR markers for use in varietal identif ication of coffee. The f indings also indicate the possible application of SSR markers in other existing cultivars available in the country. Proper identif ication is important to ensure homogeneity and increase marketability. Moreover, the application of SSRs could later be extended for marker-assisted selection of important traits, such as disease resistance, aroma, and yield. Marker-assisted selection would provide a bottom-up evolutionary approach in genetic improvement, which is more acceptable to society compared to the top-down approach of genetic engineering. ACKNOWLEDGMENT S i m p l e S eq u e n ce Re p e a t A n a l y s i s of S e l ec ted N S I C- r eg i s tered Coffee 14 The authors would like to thank the following: DOST – PCAARRD, UP OVCRD, UP Natural Sciences Research Institute, and UP Institute of Biology for the funding and support of this project; Bureau of Plant Industry in Baguio City, Benguet, Cavite State University in Indang, Cavite, and Nestle Philippines, Inc. in Malaybalay, Bukidnon for providing the NSIC-registered coffee varieties; and Professor Valentino Macanes of the Benguet State University and his staff for providing resources and information. REFERENCES Anthony F, Ber trand B, Quiros O, Wilches A, Lashermes P, Berthaud J, Charrier A . 2001. Genetic diversity of wild coffee (Coffea arabica L.) using molecular markers. Euphytica. 118(1):53-65. Anthony F, Combes MC, Astorga C, Ber trand B, Graziosi G, Lashermes P. 2002. The origin of cultivated Coffea arabica L. varieties revealed by AFLP and SSR markers. Theoretical and Applied Genetics. 104(5):894-900. Bigirimana J, Njoroge K, Muthomi JW, Gahakwa D, Phiri NA , Gichuru EK, Walyaro DJ. 2 0 1 3 . G e n e t i c d i v e r s i t y a m o n g d i s e a s e r e s i s t a n t c o f f e e v a r i e t i e s a n d c u l t i v a r s i n Rwanda based on RAPD and SSR markers. Journal of Renewable Agriculture. 1(6):106- 112. Cao EP, Constantino-Santos DM, Ramos LAP, Santos BS, Quilang JP, Mojica RM. 2014. M o l e c u l a r a n d m o r p h o l o g i c a l d i f f e r e n t i a t i o n a m o n g C o f f e a ( R u b i a c e a e ) v a r i e t i e s grown in the farms of Cavite Province, Philippines. Philippine Science Letters. 7(2):387- 397. Charrier A, Ber thaud J. 1985. Coffee: Botany, Biochemistry and Production of Beans and Beverage. In: Clifford MN, Wislon KC, editors. London: Croom Helm. p. 13-47. Geleta M, Herrera I, Monzon A , Bryngelsson T. 2012. Genetic diversity of arabica coffee (Coffea arabica L.) in Nicaragua estimated by simple sequence repeat (SSR) markers. The Scientif ic World Journal. 1-11. G i c h i m u B M , G i c h u r u E K , M a m a t i G E , N ye n d e A B . 2 0 1 4 . O cc u r r e n ce of C K- 1 g e n e conferring resistance to coffee berry disease in C. arabica v. Ruiru 11 and its parental genotypes. Journal of Agricultural and Crop Research. 2(3):51-81. Gichuru EK, Agwanda CO, Combes MC, Mutitu EW, Ngugi ECK, Ber trand B, Lashermes P. 2008. Identif ication of molecular markers linked to a gene conferring resistance to cof fee b e r r y d i s e a s e ( Co l l e t o t r i c h u m ka h a w a e ) i n Cof fe a a r a b i c a . P l a n t Pa t h o l o g y. 57:1117-1124. Janzen DH. 2009. A DNA barcode for land plants. PNAS. 106(31):12794-12797. Lashermes P, Combes MC, Rober t J, Trouslot R, D’Hont A , Anthony F, Charrier A . 1999. Molecular characterization and origin of the Coffea arabica L. genome. 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Microsatellite markers in plants and insects. Par t I: Applications of Biotechnology. Genes, Genomes and Genomics. 3(1):1-14. _____________ Daisy May C. Santos is currently working as a Science Research Specialist and as a lecturer at the Institute of Biology, University of the Philippines Diliman. She f inished her Bachelor of Science degree in Biology from the University of the Philippines Baguio and her Master of Science degree in Biology (major in Genetics) from the University of the Philippines Diliman. Her research interests include plant genetics and molecular biology. S i m p l e S eq u e n ce Re p e a t A n a l y s i s of S e l ec ted N S I C- r eg i s tered Coffee 16 Carla Francesca Besa is a graduate of BS Agricultural Biotechnology at the University of the Philippines Los Baños. She is also a member of the Alpha Chi Chapter of Phi Sigma Society, and is currently working as a Science Research S p e c i a l i s t u n d e r t h e C o f f e e G e n o m i c s P r o j e c t a t t h e P l a n t G e n e t i c s a n d Cyanobacterial Biotechnology Laboratory of the University of the Philippines Diliman. Angelo Joshua V ictoria is a graduate student at the Institute of Biology where he also works as a researcher. His research interests include molecular genetics and dabbling with bioinformatics. He also has a keen interest in foreign languages and learns German and Swedish during his time out of the lab. Dr. Ernelea P. Cao is currently Professor 12 and Director of the Institute of Biology, College of Science, University of the Philippines Diliman. She was former Deputy Executive Director of the Philippine Genome Center and was also the former Director of the Natural Sciences Research Institute, College of Science, University of the Philippines Diliman. She f inished her Bachelor of Science degree in Biology from the University of the Philippines Los Baños (major in Genetics) and her M.S. and Ph.D. in Biology at the University of the Philippines Diliman (major in Genetics). She conducted her dissertation under a sandwich program at the Department of Biology, University of North Carolina in Chapel Hill, North Carolina, U.S.A . Her research interests include plant genetics and cyanobacterial biotechnology.