Emma K. Sales

Details

Emma K. Sales

Sex: Female

Description: Sales established the first tissue culture and biotechnology laboratory in Soccsksargen for developing diagnostic tools for the identification of durian, rubber, and mango varieties.

Researches:

Article title: Molecular Evaluation of the Philippine Best Rubber Clones using Genomic-based Simple Sequence Repeats

Authors: Aldrin Y. Cantila, Rene Rafael C. Espino, Emma K. Sales

Publication title: Journal of Advanced Applied Scientific Research 1(11):9-27, August 2017

Abstract

University of Southern Mindanao Agricultural Research Center, Philippines conserved 86 rubber clones in the field germplasm. Among rubber clones, only seven were commercially released and widely utilized by Filipino rubber farmers. Nineteen genomic-based simple sequence repeats (G-SSRs) provided enough data onto evaluating molecular information of Philippine best rubber clones (five PB, one RRIM and one Philippine or Phil derived clones) along with two RRIM and Phil check clones. G-SSRs derived 72 alleles in all with means N, Ne, Ho, and PIC of 3.789, 2.284, 0.569 and 0.508 per G-SSR, respectively. Nine G-SSRs however detected 16 private alleles across rubber clones and groups that led AMOVA result to 74.37% molecular variance within clones. Private alleles in best clones could be used as molecular reference to authenticate registered clones especially in nursery farm-producing planting materials. Genetic relationship was in a range of 0.184 to 0.487 proximities, having PB217 and SMRX1 as the closest. Bayesian structure analysis on the other hand distributed clones into two groups (group 1-RRIM600, RRIM712 and RRIM901; group 2-PB217, PB235, PB260, PB311, PB330, SMRX1, UPLBPlant1 and USM1); a reference for in-depth consideration of making crosses.

Full text available upon request to the author

Article title: Genetic variation and structure of rubber population based on microsatellites

Authors: Aldrin Y. Cantilla, Rene Rafael C. Espino, Emma K. Sales

Publication title: International Journal of Biosciences 10(3), March 2017

Abstract:

Information towards genetic variation and structure of rubber leads to a proper utility of rubber clones for a varietal development. Rubber population represented by nine Asian, 10 South American and nine West African clones was described by indices for genetic variation such as number of alleles (Na), number of effective alleles (Ne), observed heterozygosity (Ho), unbiased heterozygosity (uHe), fixation index (F), Shannon's information index (I) and gene diversity (GD); and population structure such as analysis of molecular variance (AMOVA), structure analysis and principal component analysis (PCA) based on 13 microsatellites. Microsatellites derived 60 alleles in all with means Na, Ne, Ho, uHe, F, I and GD of 4.615, 2.997, 0.619, 0.686, 0.051, 1.174 and 0.647 per microsatellite, respectively. AMOVA revealed 4%, 16% and 80% genetic variation among groups, among and within clones, respectively. The initial three groups based on geographical origin were reassigned into four based on the structure analysis.PCA supported the grouping through the distribution of clones on the scatterplot's projection. PCA also detectedPC1 clones: GW5, IAN873, IRCA22, PB311, RRIM600 and USM1 as the most variable clones based on squared cosines. Microsatellite data showed a rich genetic variation is within clones and confirmed rubber is genetically heterogenous. PCA result suggested PC1 clones can be effective parents while unbiased structure grouping will serve as heterotic groups as basis for the hybridization and development of new rubber varieties.

Article title: Utilization of Simple Sequence Repeats Markers in the Genetic Characterization of Philippine Rubber, Hevea brasiliensis (Willd. ex A. Juss) Müell. Arg., Germplasm

Authors: Aldrin Y. Cantilla, Rene Rafael C. Espino, Emma K. Sales

Publication title: Philippine Journal of Crop Science 40(3), December 2015

Abstract:

Simple Sequence Repeats (SSRs) were utilized to genetically characterize rubber genotypes among 86 clones of rubber germplasm in the Philippines. Twenty-two out of 40 screened SSR primers were found to be informative with a mean of 99.09% polymorphic rate. These primers generated 119 bands with a mean of 5.41 bands per primer in a corresponding range of 100 to 491 base pairs (bp). From 119 alleles, 118 were polymorphic and 76.4 were effective. The resolving power of molecular markers measured as Polymorphism Information Content (PIC) had a mean value of 0.65. SSRs had detected a broad genetic base of 0.30 in Unweighted Pair Group Method with Arithmetic Mean (UPGMA) indicated by Jaccard’s similarity coefficient. On the other hand, five ideal groups were computed by Ad hoc statistics for Bayesian grouping as determined using the Structure 2.3.4 program with high replications and iterations of Markov Chain Monte Carlo (MCMC) and admixture model/correlated allele frequencies, respectively. SSRs have provided efficient, reliable and useful derivation of allelic variations, polymorphism information and genetic similarities. The derived ideal groups showed potential use in the decision-making process as to what clones are to be bred.

Article title: Molecular analysis of somaclonal variation in tissue culture derived bananas using MSAP and SSR markers

Authors: Emma K. Sales and Nilda G. Butardo

Publication title: International Journal of Biological, Veterinary, Agricultural and Food Engineering 8(6), 2014

Abstract:

The project was undertaken to determine the effects of modified tissue culture protocols eg age of culture and hormone levels (2, 4-D) in generating somaclonal variation. Moreover, the utility of molecular markers (SSR and MSAP) in sorting off types/somaclones were investigated. Results show that somaclonal variation is in effect due to prolonged subculture and high 2, 4-D concentration. The resultant variation was observed to be due to high level of methylation events specifically cytosine methylation either at the internal or external cytosine and was identified by methylation sensitive amplification polymorphism (MSAP). Simple sequence repeats (SSR) on the other hand, was able to associate a marker to a trait of interest. These therefore, show that molecular markers can be an important tool in sorting out variation/mutants at an early stage.

Article title: Marker-assisted introgression of opaque 2 gene into elite maize inbred lines

Authors: Efren E. Magulama and Emma K. Sales

Publication title: University of Southern Mindanao Research & Development (USM R&D) 17(2), 2009

Abstract:

Marker assisted backcrossing can greatly accelerate the introgression of modified opaque2 gene into elite inbred lines. This study aimed to transfer opaque 2 gene into elite maize inbred lines using SSR markers. Two backcross maize (white and yellow) populations were developed by crossing quality protein (QP) line with non-QP lines and subsequently backcrossed to non-QP lines until BC3 generation. Among the three SSR markers (phi 057, umc 1066, and phi 112) used for opaque2 gene selection, only SSR phi 057 marker was employed in segregating populations owing to its good discriminatory power in the survey of parental polymorphism. Using marker assisted selection, we transferred opaque 2 gene into USM elite breeding lines. Of the 26 converted QPM lines (16 white, 10 yellow) selected from the BC3F3 populations, seven lines (4 white, 3 yellow) were finally selected as quality protein inbred lines, having satisfied the minimum standard criteria for protein quality. These converted QPM lines could be used as parent lines in the development of QPM varieties.

Article title: Improvement of bananas through gamma ray irradiation

Authors: Emma K. Sales, Jorge Lopez, R Espino, N. Butardo, et al.

Publication title: Philippine Journal of Crop Science 38(2), August 2013

Abstract:

Banana is the second most widely-grown fruit crop in the Philippines and considered as the number one dollar earner and export winner. Out of the diverse types and kinds of banana, cultivar ‘Lakatan’is the most popular in the domestic and to some extent in the export market. However, no improvement and breeding activities for this crop in the Philippines has been done due to its long life cycle and sterility trait. In order to address this, mutation breeding through gamma irradiation can be an alternative solution. This research was conducted to determine the LD50 for ‘Lakatan’(shoot tips) and two variants (embryo and cell suspension), and to develop a ‘Lakatan’cultivar with improved postharvest trait. Radio sensitivity test on shoot tip cultures established that the LD50 is the 50Gy gamma ray. Mutation induction was carried out for 500 shoot tips using 50Gy gamma ray treatment. Gamma irradiated plants were evaluated for four generations. Mutations were observed in the bunch weight, number of hands per bunch, number of fingers per hand, and shelf life. Hence, the preliminary findings suggest the potential of induced mutation through gamma ray irradiation as an alternative technique in developing/generating bananas with specific trait of interest.

Full text available upon request to the author

Article title: Assessment of ploidy and genome constitution of some Musa balbisiana cultivars using DArT markers

Authors: Emma K. Sales, Nilda G. Butardo, Humberto Gomez Paniagua, Hans Jansen, et al.

Publication title: Philippine Journal of Crop Science 36(1), April 2011

Abstract:

Musa cultivars. Likewise, dendrogram constructed by Mega version 4 clearly separated the BB/BBB cultivars from AA/AAA and AAB cultivars. Moreover, the dendrogram showed that most of these putative BBB cultivars grouped as a separate cluster from the AAA and AAB groups. Bootstrapping all the more enhanced these groupings, and strongly supported the existence of a pure Balbisiana group. These confirmed the Asian claims of the triploid B nature of the Philippine ‘Saba’group. However, two cultivars namely ‘Inambak’and ‘Pondol’(assumed to be of the BBB type) formed a cluster with the AAB (cv ‘Tindok and ‘Latundan’). This suggests therefore, that these two Phil. Musa cultivars could be AAB unlike the rest of the cultivars claimed to be triploid B.

Article title: Durian marker kit for durian (Durio zibethinus Murr.) identity

Authors: Emma K. Sales

Publication title: International Journal of Biological, Biomolecular, Agricultural, Food and Biotechnological Engineering 9(5), 2015

Abstract:

Durian is the flagship fruit of Mindanao and there is an abundance of several cultivars with many confusing identities/ names. The project was conducted to develop procedure for reliable and rapid detection and sorting of durian planting materials. Moreover, it is also aimed to establish specific genetic or DNA markers for routine testing and authentication of durian cultivars in question. The project developed molecular procedures for routine testing. SSR primers were also screened and identified for their utility in discriminating durian cultivars collected. Results of the study showed the following accomplishments: 1. Twenty (29) SSR primers were selected and identified based on their ability to discriminate durian cultivars, 2. Optimized and established standard procedure for identification and authentication of Durian cultivars 3. Genetic profile of durian is now available at Biotech Unit Our results demonstrate the relevance of using molecular techniques in evaluating and identifying durian clones. The most polymorphic primers tested in this study could be useful tools for detecting variation even at the early stage of the plant especially for commercial purposes. The process developed combines the efficiency of the microsatellites development process with the optimization of non-radioactive detection process resulting in a user-friendly protocol that can be performed in two (2) weeks and easily incorporated into laboratories about to start microsatellite development projects. This can be of great importance to extend microsatellite analyses to other crop species where minimal genetic information is currently available. With this, the University can now be a service laboratory for routine testing and authentication of durian clones.

Article title: SSR markers for mango (Mangifera indica L.) cultivar identification and genetic characterization.

Authors: Emma K. Sales and Nilda G. Butardo

Publication title: Philippine Journal of Crop Science 42(3), 2017

Abstract:

The study identified SSR markers that can discriminate the different mango cultivars in the Philippines. It was conducted to select markers that are specific for Philippine Carabao Mango that can be used as a diagnostic kit for true to type variety. Two hundred (200) mango samples were collected and evaluated while 49 SSR primers were screened for their utility in assessing the identity of the said mango cultivars. Out of these 49 primers, 29 primers were selected for further analysis. Based on the generated molecular data, e.g. allele size, fragments amplified, polymorphic information content (PIC) and power of discrimination, the 29 primers were able to discriminate carabao cultivars over that of other cultivars sampled. An arbitrary genotypic assignment was also established to further show the differences among the cultivars/samples evaluated. The results show that the primers selected can be used to validate the authenticity of the carabao strain as labelled.

Full text is available upon request to the author

Article title: Diversity analysis of maize inbred lines using SSR markers

Authors: Emma K. Sales, E.E. Magulama, N.G. Butardo, R.C. Cadungog, et al.

Publication title: Philippine Journal of Crop Science 29, 2004

Abstract:

With the assistance of the Asian Maize Biotechnology Network (AMBIONET) and DA-BAR, a molecular biology laboratory was established at the University of Southern Mindanao in 2002. National Staff was trained to strengthen the university's capability in the application of molecular techniques for crop improvement. The authors initiated the DNA fingerprinting of Philippine publicly derived maize inbred lines for local as well as region-wide diversity analyses. Initially, 87 Philippine inbred lines were fingerprinted using SSR markers and standard protocols from AMBIONET. Analysis at 21 SSR loci, however, revealed that most of the lines had 20% heterozygosity. These lines were recommended for further purification and are presently in the second selfing generation. In another study aimed at the introgression of the opaque 2 allele to elite local lines and development of quality protein maize (QPM) cultivars, 21 QPM lines (7 yellow and 14 white maize) were obtained from CIMMYT and analyzed using 40 SSR markers. SSR data of 8 QPM lines, remaining after 13 white lines 20% heterozygosity were excluded, were merged with a data set of 16 QPM lines from AMBIONET-Vietnam and with a regional dataset of 102 inbred lines from the AMBIONET Service Lab that included 10 lines from the Philippines as well as lines from various tropical and temperate countries. Error rates (ER) were calculated between laboratories and marker data having less than 10% ER were combined. By cluster analysis, the 24 QPM lines were seen in at least 7 different groups of the regional dataset, which suggests that this set of QPM lines are genetically diverse. These results are useful as a guide for selecting crosses with potential superior combining ability. The authors continue to analyze the diversity of Philippine inbred lines and eventually combine it with data sets of other lines that are relevant to local breeders.

Full text is available upon request to the author

Article title: RAPD markers for genetic analysis and classification of Musa B genome

Authors: Emma K. Sales and RRC Espino

Publication title: Philippine Agricultural Scientist 87(2), 2004

Abstract:

Full text is available upon request to the author

Article title: Evaluation of USM Biotech DNA extraction method for selected agricultural crops in comparison with existing methods

Authors: Monalyn Marimpoong and Emma K. Sales

Publication title: Journal of Agricultural Research, Development, Extension and Technology 1(1), 2019

Abstract:

Different DNA extraction protocols have been used in plant DNA extraction. However, no comparative analysis has been done to determine their efficiency, cost effectiveness and time requirement for the extraction. Three (3) established protocols and the USM Biotech modified protocols were used in this study. It aimed to evaluate the efficiency of the four DNA extraction protocols in terms of DNA yield, purity and processing time; to determine and compare the cost of sample analysis per protocol and to assess which protocol is universally applicable in extracting DNA of selected agricultural crops (banana, cacao, durian, mango and rubber). The experiment was carried out in 4X5 factorial arranged in Complete Randomized Design (CRD), Factor A as protocols and Factor B as crops used. Results of the study showed that among four (4) protocols, the protocol developed by Ferdous et al.(2012) was the most cost effective. It was the least expensive and fastest method of extracting DNA resulting to high genomic DNA yield. Likewise, University of Southern Mindanao (USM) Biotech modified protocol was found to be another efficient, economical and effective method of extracting DNA with sufficient amount of DNA yields. The protocol developed by Ray et al.(2016) produced the highest DNA yield; however, it was the most time-consuming method among the four (4) protocols. The Diversity Array Technology (DArT) protocol on the other hand, was the most expensive method among the four protocols because it required the use of expensive reagents and liquid nitrogen.

Full text is available upon request to the author

Article title: Molecular Evaluation of the Philippine Best Rubber Clones using Genomic-based Simple Sequence Repeats

Authors: Aldrin Yanong Cantila, Rene C. Espino, Emma K. Sales

Publication title: Journal of Advanced Applied Scientific Research 1(11), 2017

Abstract:

To date there are only nine best rubber clones in the Philippines with little knowledge about its molecular information. Nineteen genomic-based simple sequence repeats'(G-SSRs) good genome coverage had provided enough data in evaluating the Philippine best rubber clones (five PB, three RRIM and one Philippine or Phil derived clones) along with two Phil check clones. G-SSRs derived 72 alleles in all with means N, N e, H o, and PIC of 3.789, 2.284, 0.569 and 0.508 per G-SSR, respectively. Nine G-SSRs however detected 16 private alleles. AMOVA also revealed 6.37%, 19.26% and 74.37% molecular variance between groups, between and within clones, respectively, with corresponding F ST of 0.064. While genetic proximity had a range of 0.184 to 0.487 with corresponding neighborhood-joining tree suggested no similar clones, three groups (PB, RRIM and Phil groups) had delta K of two based on Bayesian structure analysis. Group 1 had RRIM600, RRIM712 and RRIM901 while group 2 had PB217, PB235, PB260, PB311, PB330, SMRX1, UPLBPlant1 and USM1. G-SSRs are suitable for extracting molecular information and distinct alleles, which can be used as reference to authenticate the best rubber clones in the Philippines. Bayesian structure analysis on the other hand assured better resampling strategy and found admixtures. Furthermore, rubber is highly structured on an individual level that indicates these clones should be utilized to broaden genetic variability of the Philippine rubber.

Full text available upon request to the author

Article title: Involvement of genomics in rubber breeding

Authors: Pascal Montoro, K. Annamalainathan, Thakurdas Saha, Sigit Ismawanto, et al.

Publication title: Agricultural Research for Development, 2017

Abstract:

The 5-year strategic plan of IRRDB has highlighted that studies on physiology, molecular biology and biotechnology are involved in a range of research areas dealing with the sustainable rubber production (productivity, adaptation, NR quality, development of new products and environmentally friendly practices), the preservation of natural resources (Hevea genetic resources, biodiversity in rubber plantation), and the reduction of greenhouse gas emissions (carbon sequestration). These researches have been leading to a better understanding of important agronomic traits and to develop new high yielding rubber clones with better tolerance to biotic and abiotic stress in a context of climate change. Molecular mechanisms involved in NR production and plant defence have complex regulations requiring global research approach through omics technologies. Application of these technologies has led to decipher molecular bases of important agronomic traits and to assist breeding programmes.

The main objective of the CATAS-IRRDB workshop on Rubber genomics and molecular genetics was to share knowledge and experiences in genomics, transcriptomics, proteomics, metabolomics, molecular genetics and breeding among researchers and scientists especially from the IRRDB member countries. Updated information on genomics and transcriptomics in rubber and genome-based applications has been delivered by keynote speakers, and a round table discussion was conducted to identify topics for developing international initiatives.

Full text available upon request to the author

Article title: Potential Banana cv 'Lakatan' Somaclones Induced by Long Culture Period and High 2, 4-D Concentration

Authors: Emma K. Sales, Harem R. Roca, Nilda G. Butardo

Publication title: Philippine Journal of Science 145(2), June 2016

Abstract

Out of these 2,040 plants, 40 somaclones were selected based on their better performance compared to the untreated plants (control). Results showed that prolonged subculture and addition of high concentration of 2, 4-D produced both positive and negative variations. Positive variation was exhibited by heavier bunch weight, earlier flowering, longer shelf life and a larger number of hands, which translate into increased income. Negative variation, on the other hand, included dwarfism, delayed flowering and a lesser number of hands.

Article title: Applications of in vitro methods for the multiplication of new planting material

Authors: Pascal Montoro, Keng See Chow, Philippe De Groote, Nor Mayati, et al.

Publication title: Agricultural Research for Development, 2014

Abstract:

Hevea clones are propagated by grafting scion clone on seedling rootstocks. Cutting and microcutting were long supposed to provide plant material with poor root system. Thanks to the French-Indonesian research collaboration and development in China, microcutting is now reconsidered as useful technology for Hevea clonal propagation and production of rootstocks. Somatic embryogenesis is cost-effective clonal propagation method that has been maintained in China, India, France and Malaysia for more than 30 years. This long-term investment led to applications on genetic engineering and large-scale productions of in vitro plants in China by CATAS. In the recent somatic embryogenesis is an essential factor for successful horticultural multiplication and improving the vigour of planting material. Using rejuvenated in vitro plants from somatic embryogenesis, grafting and cutting led to the multiplication of thousands plants in Nigeria and Ivory Coast by CIRAD-Michelin-SIPH and University of Gand-Socfinco consortium, respectively. Effect of clonal root system and somaclonal variation must be taken into consideration. Evaluation of this new planting material remains necessary for further recommendations. Given the low rubber price, other latex by-products should also be considered to attract farmers to the cultivation of rubber. Natural organic compounds as well as new recombinant proteins could lead to a better attractiveness to stakeholders and a better acceptance of genetically modified rubber plants.(Texte intégral)

Full text available upon request to the author

Title: International rubber research and development board biotechnology group annual report 2010

Authors: Pascal Montoro, Keng See Chow, Zhe Li, Thitaporn Phumichai, et al.

Publisher: International Rubber Research and Development Board, 2011

Brief description:

The increasing demand for natural rubber calls for improved productivity and extension of rubber growing areas to non-traditional in most rubber growing countries. Biotechnologies play a role in breeding and also in understanding the various plant biological processes involved in plant development and defence against biotic and abiotic stresses. Research advances reported in 2010 confirm this trend

Article title: Improved postharvest technologies in marang

Authors: Emma K. Sales, N.A. Turnos, J.G. Tangonan, P.B. Barit

Publication title: Philippine Journal of Crop Science 36, 2011

Abstract:

The research was conducted to survey , assess, and improve current postharvest practices in marang, identify the right maturity indices, exploit existing strategies in regulating ripening, develop processing technologies and products to enhance the profitability of marang production through value-adding activities. A benchmark survey on the current postharvest practices was carried out to assess the gaps to reduce postharvest losses. Ripening of the fruits was manipulated using packaging materials, ehtylene inhibitors and ethylene inducers. Possible processing technologies were also explored to develop marang products (minimally-processed and processed). Results showed that the right stage of maturity for harvesting of the commercially-available 'Evergreen' and 'Brown' cultivars in the local markets is 80-90 days from fruit set. To minimize losses, harvesting should be done using modified tools. Storability and shelf life can be prolonged using appropriate packaging materials and cold storage. Regulation of ripening can be attained through the following: washing and covering the fruit with wet sack or cloth were found to be faster, easier and economical ripening techniques than using ripening agents e.g. calcium carbide, kakawati [Gliricidia sepium] leaves and other ripening materials. On the other hand, ethylene scrubber and packaging in a closed polyethylene bag or kept under modified atmosphere (MA) condition delayed the ripening. For minimally processed marang pulps, addition of an antioxidant, metabisulfite, when packed in either polyethylene bag (Ziploc),. vacuum sealed bags or styrofoam delays and minimizes browning. Moreover, with these treatments, taste was rated acceptable. Cold storage at 0 and 4 deg C also gave an acceptable appearance of the pulps. These results proved that it is possible to bring minimally processed and processed marang products from Mindanao to other parts of the Philippines without affecting its sensory quality. The following marang products were developed: frozen pulps or arils, puree, concentrates, jam, conserve, vacuum-fried, dehydrated, spray-dried, ice cream, and other by-products.

Full text available upon request to the author

Article title: A method of increasing the efficiency of finding productive crosses in maize

Authors: Efren E. Magulama and Emma K. Sales

Publication title: Maize for Asia: Emerging trends and technologies, 2008

Abstract:

Innovative methods in reducing the cost while increasing efficiency of breeding work are of great help to maize breeders. This study aimed to evaluate the usefulness of cluster-based mating using SSR and qualitative morphological markers in finding productive crosses (PC) in maize. In this study we used four datasets that consisted of 192 crosses derived from four sets of three diallel and a design II experiments in which three sets were inbred crosses and a set of variety crosses. The four sets of crosses including the check varieties were evaluated for yield in alpha lattice design with three replications. We used 39, 40, and 30 SSR markers to establish grouping of inbred lines in set A, B, and C, respectively. For set D, we used 19 agro-morphological traits in grouping the parent varieties. Using the four data sets, the results showed that inter-group crosses had greater number of PCs than those in intra-group crosses. Sixty-four percent (64%) of the PCs came from the inter-group crosses in set A. In set B, C, and D all the PCs were come from inter-group crosses. Further, clustered-based mating appeared more efficient in identifying productive crosses in maize by 12% than the non clustered-based mating, thereby reduced the cost of extensive field crosses and tests.

Article title: Genome and ploidy confirmation of Phil. Musa balbisiana cultivars based on flow cytometry analysis and DArT markers

Authors: Emma K. Sales, N.G. Butardo, T.C. Solpot, J.B. Ranis

Publication title: Philippine Journal of Crop Science 2008

Abstract:

The major constraint in banana and plantain research is the problem on proper identification and classification due to the endless range of names and synonyms for cultivars. In terms of Musa balbisiana taxonomic classifications, there is an ongoing controversy regarding the existence of pure edible balbisiana cultivars endemic to the Philippines. Asian and Philippine Musa workers claimed that Philippine 'saba' group of the triploid B while American and European Musa workers denied the existence of such and consistently claimed that they are a hybrid of A and B genomes. The objective of the study was to determine if DArt analysis can efficiently verify the genome constitution of the Musa cultivars in question. To shed light on this, 24 Philippine Musa cultivars were subjected to flow cytometry analysis for ploidy confirmation and DArt analysis for molecular characterization. Results of the flow cytometry confirmed the triploidy of the Philippine 'Saba' group while the molecular data showed that some DArt markers were specific for B genome such that these markers can identify the cultivars with B genome regardless of the presence of A genome. Hence these markers can be used to establish genome identity of the Musa cultivars. Likewise, dendrogram constructed by Mega version 4 clearly separated the BB/BBB cultivars from AA/AAA and AAB cultivars. Moreover, the dendrogram showed that most of these putative BBB cultivars, grouped as a separate cluster from the AAA and AAB groups. Bootstraping all the more enhanced supports the existence of pure Balbisiana group and might confirm the Asian claims of the triploid B nature of the Philippine 'Saba' group. However, two cultivars namely 'Inambak' and 'Pondol' (assumed to be of the BBB type) formed a cluster with the AAB (cv 'Tindok' and 'Latundan'). This suggests therefore, that these two Philippine Musa cultivars could be AAB unlike the rest of the cultivars claimed to be triploid B.

Full text available upon request to the author

Article title: Analyses of genetic diversity and heterotic patterns in maize inbred lines as revealed by SSR markers

Authors: Emma K. Sales, E.E. Magulama, Cotabato Philippines Kabacan, RC Cadungog, et al.

Publication title: Philippine Journal of Crop Science 30, 2004

Abstract:

Knowledge of genetic diversity combining ability, and heterotic patterns of maize germplasm is of great important to maize breeders. The study was undertaken to (a) assess the genetic diversity of 11 yellow maize inbred lines using SSR markers, and (b) determine the combining ability of the 11 yellow maize inbred lines and (c) select inbred lines that could b used as parent materials for heterosis breeding and for synthetic variety formation and (d) verify the grouping of inbred lines based on molecular markers with field performance of hybrid progeny. Eleven yellow maize inbred lines were used and crossed in diallel fashion to form 55 single crosses. The 55 crosses were tested in field and arranged in alpha lattice with three replications. Each entry was planted to three 3-m row plots. Recommended cultural management practices were strictly followed. Using a 39 informative SSR markers, cluster analysis classified 11 lines into four groups consisting of GI: 4 lines, GII- 2 lines, GIII- 3 lines, and GIV-1 line. Of the 14 superior single crosses to hybrid check, 12 crosses were coming from crossing inbred lines belonging to different groups and only 2 crosses from the same group. Differences in crosses were mainly due to specific combining ability effects constituting 78% for yield, 81% for plant height, 69% for silking date, and 78% for moisture content, and 55% for kernel number per year. This indicates the non-additive effects conditioning the expression ofthese traits gathered. Five inbred lines were identified as possible parents for synthetic variety development, while ten single cross hybrids were identified as promising single hybrids. They gave a yield potential of 8 to 9 t/ha which were relatively better than the hybrid check variety. The study also showed that grouping the lines prior to making crosses using molecular markers and subsequent crossing using factorial mating design were more efficient than the conventional diallel mating in identifying and selecting superior hybrids. The results imply that the use of molecular markers can greatly help in reducing the number of crosses, thereby, reducing the cost and time in making crosses and field testing.

Full text available upon request to the author

Article title: RAPD [random amplified polymorphic deoxyribonucleic acid] markers for genetic analysis and classification of Musa B genome

Authors: Emma K. Sales, R.R.C. Espino

Publication title: Philippine Agricultural Scientist 87(2), 2004

Abstract:

Random amplified polymorphic deoxyribonucleic acid (RAPD) markers were determined to classify the B genome of Musa germplasm collections. Optimization of several parameters for DNA amplification and RAPD analysis was also done to determine RAPD reaction conditions applicable for Musa cultivars in the B genome. A reaction mixture of 35 ng DNA, 2.5 m MgCl2, 1 unit/u L Taq DNA pol, 1 microM dNTPs, 0.50 ng RAPD primer and 1XPCR buffer was found optimum for DNA amplification. The RAPD polymerase chain reaction (PCR) temperature cycle profile developed for rice was similarly found applicable for the Musa cultivars evaluated. Moreover, a random ten-over primer with 60%-70% G + C obtained good RAPD band pattern. Specifically, primers OPH-03 and OPI-16 detected three DNA markers which were present in all BB/BBB cultivars. These DNA markers (0.560 kb marker detected by primer OPH-03 and 0.836 and 0.690 kb markers detected by primer OPI-16) were therefore used to determine the genome constitution of three Pinatubo collections and Palawan IX. Cluster analysis using the RAPD markers data showed that the balbisiana cultivars were different from acuminata cultivars as represented by `Bungulan' and `Lakatan', respectively. Furthermore, the Pinatubo collections were shown to be of the balbisiana group. However, the genome classification of the Palawan collections was not clear because of inconsistencies in clustering.

Full text available upon request to the author

Article title: Use of molecular markers for heterotic grouping of publicly developed maize inbred lines in the Philippines

Authors: Emma K. Sales, E.E. Magulama, N.G. Butardo

Publication title: Philippine Journal of Crop Science 28, 2003

Abstract:

Ninety-three maize inbred lines developed at USM, UPLB [Philippines Univ. Los Banos, Laguna, Philippines], and IES were assayed for polymorphism at 40 SSR [simple sequence repeats] marker loci using polyacrylamide gel electrophoresis (PAGE). The objectives were: to screen a set of stable and repetitive SSR markers for DNA fingerprinting or heterotic grouping and exploit the heterotic patterns among domestic inbred lines. Two hundred six alleles were identified and served as raw data for estimating genetic similarities among these lines. A cluster analysis placed the inbred lines in six clusters that is suggestive of the major heterotic group for Philippine maize. Specific alleles exclusive for certain lines were also detected. These can be used as a unique fingerprint for such inbred and therefore might be a useful marker to distinguish a character trait exhibited by that particular line. The study confirms the utility of polymerase chain reaction (PCR) based markers such as SSRs for measuring genetic diversity, for assigning lines to heterotic groups and for genetic fingerprinting

Full text available upon request to the author

Article title: Genetic diversity in the B genome of Musa germplasm assessed by RAPD [random amplified polymorphic DNA] technique

Authors: Emma K. Sales, A.A. Barrion, M.S. Mendioro, M.L.O. Cedo

Publication title: Philippine Journal of Crop Science 27, 2002

Abstract:

The genetic diversities and relationships among 19 Philippine Musa cultivars based on the DNA fingerprints were investigated. DNA of the 19 cultivars extracted through modified CTAB protocol were analyzed using random amplified polymorphic DNA(RAPD) technique. A total of 118 random primers were used in the analysis. Of these primers, 41 were identified useful in distinguishing the 19 Musa cultivars. Primers J-04, S-02, and AJ-01 clearly showed genetic divergence in the 19 cultivars as evidenced by the different band, detected three DNA markers, which are present in all the BB/BBB cultivars. Through these DNA markers (0.560 kb marker detected by primer OPH-03 and 0.836 kb markers detected by Primer OP1-16 the genomic sequence constitution of six Musa collections was determined. Cluster analysis at specific similarity coefficients revealed that the 11 M. balkesiana cultivars evaluated are different and distinct from M. acuminata cultivars as represented by 'Bungulan' and 'Lakatan'. It also suggests that the three Pinatubo collections are of the BB/BBB genome while the three Palawan collections might be of the AA/AAA type. Some degrees of variability were also detected among the BB/BBB cultivars. This study demonstrated that RAPD could be used for Musa germplasm characterization, identification and fingerprinting. It is useful in establishing phenetic and phytogenetic relationships as well as genomic constitution of Musa cultivars. It can also be used in detecting genetic diversities particularly in the B genome groups of Musa cultivars.

Full text available upon request to the author

Article title: Initiation of meristematic buds of benzyl amino purine on banana (Musa spp.) cultivars for somatic embryogenesis

Authors: Emma K. Sales, L.R. Carlos, R.R.C. Espino

Publication title: Philippine Agricultural Scientist 84(1), 2001

Abstract:

No abstract

Full text available upon request to the author

Article title: Assessing Genetic Diversity of Asian-based Rubber Populations using SSR and Multivariate Statistics in the Philippines

Authors: Aldrin Y. Cantila, Rene Rafael C. Espino, Emma K. Sales

Publication title: Journal of Biology, Agriculture and Healthcare 6(16), 2016

Abstract:

Assessing genetic diversity of rubber populations is important for the effective utilization of rubber genetic resources. Diversity indices such as number of alleles (Na), observed heterozygosity (Ho), gene diversity (GD), polymorphism information content (PIC) and power of discrimination (PD) along with multivariate statistics such as principal component analysis (PCA) and clustering analysis were used in the study. Twenty-two SSR markers had means 5.09 Na, 0.579 Ho, 0.677 GD, 0.643 PIC and 0.785 PD for 63 rubber clones comprised of 34 Indonesian and 29 Malaysian clones. Malaysian subpopulation had 3.59 Na per clone greater than Indonesian subpopulation of 2.97 Na per clone. PCA detected 66.08% total variation for eight principal components (PCs). PC1, PC2 and PC3 contributed 13.24% variation (v) with 2.91 eigenvalue (e), 10.2% v with 2.24 e and 8.86% v with 1.95 e, respectively. Clustering analysis revealed 0.237 genetic similarity and ten clusters for all clones. Clusters will be the basis for making more genetically diverse hybrids while PC1 member clones will be the basis for considering genetically broad base parent. The high genetic diversity found in the Asian-based rubber populations and complementing results of multivariate statistics can optimize the selection and breeding of rubber genetic resources in the Philippines.

Papers Presented

Article title: Functional genomics-and bioinformatics-assisted gene marker development for economically important traits in cacao

Authors: E.A. Barlaan, Emma K. Sales, A.C. Laurena

Publication title: IV International Symposium on Molecular Markers in Horticulture 1203, 2017

Abstract:

Varietal improvement is needed in cacao for increased yield, resistance to pests and diseases, and other economically important traits. To facilitate marker-assisted breeding and selection programs, molecular markers are essential tools. Through functional genomics and bioinformatics, gene markers were generated using two approaches: genome filtering using methylation-sensitive restriction enzymes and gene mining in the cacao genome sequence database. In the genome-filtering method, genomic DNA of a number of cacao cultivars was digested with methylation-sensitive restriction enzymes for ligation, amplification, cloning, sequencing and bioinformatics analysis. From 215 library samples, a total of 578 putative simple sequence repeat (SSR) loci were predicted from the sequences, with 271 perfect loci and 307 ambiguous. The unique and perfect SSRs with primers were subjected to homology and gene ontology analysis. These SSRs were involved in 11 molecular functions, 11 cellular components, and 25 biological processes. In the gene-mining approach, genome sequences were downloaded from the database for bioinformatics analysis. Sequences were analyzed for de novo annotation, SSR prediction and deduplication, SSR marker and primer development, homology and gene ontology. A total of 4897 putative SSR locus regions were predicted from the coding DNA sequence regions of the cacao genome, with 3423 perfect loci and 1474 ambiguous loci. The unique and perfect SSRs with primers were involved in seven molecular functions, six cellular components, and 17 biological processes. SSR markers for genes of interest from both approaches were selected for amplification to assess polymorphism among cacao cultivars.

Full text available upon request to the author

Article title: Functional Genomics and Bioinformatics-Assisted Gene Marker Development for Economically Important Traits in Rubber

Author: Emma K. Sales

Conference title: CRRI & IRRDB International Rubber Conference 2016, Siem Reap Cambodia

Abstract:

The increasing demand of natural rubber products by the automotive, industrial, construction and medical supplies industries has renewed interest in revitalizing rubber production in the Philippines. This can be achieved by increasing the hectarage planted to rubber and the use of clones with improved traits such as high yield and pest and disease resistance. Towards attaining such goals, research was carried out in the areas of functional genomics and bioinformatics to develop gene markers for important agronomic traits. A study was undertaken to (a) develop and establish gene markers and ESTs for a library database in Hevea brasiliensis,(b) sequence gene specific regions in rubber using NGS technology,(c) construct an exome database for at least two H. brasiliensis genotypes with extreme phenotypic traits,(d) search for genespecific markers from the exome database and (e) design PCR primers for gene-specific sequences and validate these primers for trait association. Seven NSIC recommended varieties (PB260, RRIMM600, PB330, PB217, PB311, USM1, and PR107) were selected as test samples. Genomic DNAs of these samples were isolated for library preparation using rare cutter restriction enzymes for subsequent cloning. A total of 133 colonies were sequenced and subjected to BLAST and gene ontology analysis. DNA sequences were obtained that bore similarities to genes involved in 18 biological processes, 8 cellular components and 9 molecular functions. SSR primers were developed and selected for preliminary testing and utilization of differential banding patterns among the seven NSIC recommended clones.

Article title: Mutation induction in Philippine bananas cv'Lakatan'thru gamma ray irradiation

Authors: Emma K. Sales and R.R.C. Espino

Publication title: International Symposium on Induced Mutation on Plants (ISIM), 2008

Abstract:

Banana is the most important crop grown in the Philippines. Among the cultivars grown, 'Lakatan' is the most popular and commands a higher price in the local market. Despite high production, losses due to over ripening, bruising and short shelf life is one of the major constraints in a successful banana industry. The use of chemicals for delayed ripening however, remains an issue of concern due to economic and organic products advocacy. Thus, development and generation of new improved 'Lakatan' cultivar through gamma ray irradiation was carried out. Mutation was induced in 'Lakatan', a popular Philippine cultivar using gamma ray irradiation. Radio sensitivity was established at 50Gy. Morphological, cytological and molecular analysis done showed significant variations between the irradiated samples and the non-irradiated plants. In terms of morphological parameters, gamma ray irradiation affected leaf traits resulting to increased leaf width, leaf length, and number of leaves. Stem girth on the other hand was significantly reduced. Cytological observations showed that gamma irradiation increased the epidermal width, leaf thickness and size of stomates but reduced the number of stomates. For post harvest attributes, gamma irradiation prolonged the shelf life of banana fruits from 11 days to 14 days. Molecular analysis showed that some markers (RAPD and AFLP) were able to detect unique bands in samples irradiated with 50Gy while the SSR markers did not detect any band difference between the irradiated samples and the control.

Full text available upon request to the author

Article title: Application of DNA molecular markers to maize improvement in the Philippines

Authors: Emma K. Sales, P.S. Guzman, A.D. Manceras, E.E. Magulama, A.M. Salazar

Conference title: Annual Philippine Corn Symposium and Planning Workshop, Pansol, Calamba City (Philippines), 15-17 Jan 2003

Abstract:

The Molecular Biology Laboratory at USM [University of Southern Mindanao, Kabacan, Cotabato, Philippines] was established through the DA-BAR [Department of Agriculture - Bureau of Agricultural Research, Diliman, Quezon City, Philippines] IDG Fund in partnership with AMBIONET. This brings new technology to USM in reaching its goal of state-of-the art research. This project aims to: enhance the capability of public maize breeding institutions in the Philippines through biotechnology tools and develop maize germplasm through biotechnology for resource to assess the genetic diversity of publicity developed inbred lines in the country; develop quality protein maize (QPM) hybrids adapted in the Philippines; transfer the QPM allele (opaque 2) to an elite Philippine inbred through marker assisted backcrossing; contribute to the CIMMYT [Centro Internacional de Mejoramiento de Maiz y Trigo, Mexico] led drought tolerance mapping project by evaluating RILS under drought conditions. Training of USMARC research staff was conducted at USMARC Genetics and Molecular Biology Laboratory through AMBIONET for familiarization with different equipment and their uses. Likewise, lectures and demonstrations on DNA extraction, RNase treatment, DNA reprecipitation, DNA resuspension, agarose gel electrophoresis, Simple Sequence Repeats - Polymerase Chain Reaction (SSR-PCR), Denaturing Polyacrylamide Gel Electrophoresis (PAGE), Silver Staining, Scoring DNA bands and Data Analysis were also conducted. Diversity Analysis of 43 inbred lines from UPLB, 11 from Ilagan Experiment Station (IES) and 45 from USMARC, USM inbred lines is ongoing. Screening of CIMMYT drought parental lines and field evaluation of F1 segregating population will be done. Moreover, development of QPM cultivars and molecular assisted (MAS) introgression of opaque alleles to an elite Philippine inbred line will be done. So far, 55 CML's and 5 QPM lines from CIMMYT Mexico have just arrived and are ready for planting. Likewise, incorporation of opaque 2 gene to a Philippine DMR line will also be done by using 2 markers in the backcrossing work.

Full text available upon request to the author

Article title: Optimization of the random amplified polymorphic DNA (RAPD) protocol for Musa cultivars in the B genome

Authors: Emma K. Sales, R.R.C. Espino, A.A. Barrion, M.S. Mendioro, et al.

Conference title: Asian Agriculture Congress, Manila (Philippines), 24-27 Apr 2001

Abstract:

Although RAPD band patterns are easily obtained, several parameters of the amplification reaction need to be optimized in order to produce reproductible results. Optimization of several parameters for DNA amplification and RAPD analysis was undertaken to determine RAPD reaction conditions applicable for Musa cultivars in the B genome. Overall, the results suggest that a combination of 35 ng DNA, 2.5 m MCl2, 1 unit uL Tag DNA pol, 1 uM dNTPs, .50 ng RAPD primer and 1X PCR buffer is optimum. Results also suggest that the RAPD PCR temperature cycle profile developed by Zeng et al. (1995) for rice is applicable in the Musa cultivars evaluated. Moreover, a random ten-mer primers with 60-70% G + C is needed to obtain good RAPD band pattern.

Full text available upon request to the author

AUXILIARY MENU

GOVPH

Emma K. Sales