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December 2020, Vol. 103 No. 4

One of the most popular studies of coffee is in its aroma, together with its fatty acid content. The aroma of the coffee is composed of so many volatile compounds (Mondello, 2005). In this paper, the composition of volatile compounds of different varieties of coffee were analyzed using Headspace-Gas Chromatography-Mass Spectrometry. Data comparisons were made to determine the differences in the aroma profiles of each sample based on variety and roasting degree.

{Photographs courtesy of K.P. Marquez, Institute of Chemistry, University of the Philippines Los Baños, and Ms. Claire Sarah Magante}


Overexpression of Maize (Zea mays L.) Malate Dehydrogenase (ZMMDH) in IR64 Rice (Oryza sativa L.) Leads to Altered Carbohydrate Metabolism as Revealed by Transcriptomics and Metabolite Analysis
Galang, Emmanuel T., Legaspi, Charisse Leanne B., Lin, Hsiang-Chun, Quick, William Paul G., Villa, Neilyn O., Duka, Ivan Marcelo A.

Received: 20 September 2019/ Revised: 28 September 2020/Accepted: 10 November 2020

Malate dehydrogenase (MDH) (EC is among the major genes identified by the C4 Rice Consortium to play an important part in inducing C4 photosynthesis in a C3 rice system. Despite the vast information on the biochemical characteristics of the enzyme, the effects of engineering maize (Zea mays L.) MDH on IR64 rice (Oryza sativa L.) for the induction of the C4 pathway was never thoroughly studied in the light of transcriptomics. Two transformed IR64 rice events conferring overexpression of maize MDH (ZmMDH), namely MDH 22 and MDH 48, were analyzed using transcriptome analyses to assess differential expression of endogenous genes due to MDH overexpression. Sugar metabolite quantification was also conducted to identify changes in the concentration of simple and complex carbohydrates against the wild type (untransformed) IR64 rice. Transcriptome analyses revealed 301 differentially expressed genes in transformed rice with significant enriched effects in the downregulation of simple carbohydrate degradation. This result implies lower degradation of sucrose, fructose and glucose content as eight strongly associated genes for this function were downregulated. Carbohydrate characterization revealed significant differences in terms of simple and complex sugar content in MDH 22 and MDH 48, respectively, which aligned with the results of the transcriptome analyses. The resulting phenotype of the MDH-transformed lines indicate an increase in plant height and biomass which can be attributed to the association of growth due to the rapid conversion of simple sugars to starch and traced further to the implied change in the rate of simple sugar degradation. The study successfully established the effect of maize MDH in rice based on its transcriptome, sugar content, and phenotype.

Phytochemical Properties, Antioxidant Activities, and Cytotoxicity of Ethanolic Bran Extracts from Philippine Pigmented Rice Cultivars
Rodel M. Bulatao, John Paulo A. Samin, Rochelle C. Huliganga, Reniel P. Tubera, Mark Aldren M. Feliciano and Cesar V. Ortinero

Received: 04 November 2019/ Revised: 29 October 2020/Accepted: 10 November 2020

Antioxidants are widely recognized for their immune-enhancing and disease-preventing properties. However, there are some reports that certain synthetic antioxidants can pose carcinogenic effects on human cells. Therefore, it is vital to look for an alternative source of natural, effective, and safe antioxidants for human consumption. In this study, ethanolic extracts of six pigmented rice bran samples were evaluated for their phytochemical properties, antioxidant activities, and cytotoxicity against normal human blood lymphocytes (NHBL). Results showed that the pigmented rice bran extracts had high total phenolic (70.1–178.4 mg.g-1 gallic acid equivalent), flavonoid (123.3–378.0 mg.g-1 rutin hydrate equivalent), and anthocyanin (0.8–152.5 m.g-1 cyanidin-3-glucoside) content. They also had strong antioxidant activities that ranged from 116.4 to 461.7 mg.g-1 trolox equivalent (TE) for ferric reducing antioxidant power (FRAP) and 85.4–367.7 mg.g-1 TE for 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation scavenging activity (ABTS-RCSA). All sample extracts showed low levels of effective concentration (EC) at 50 (EC50) (11.6–30.3 mg.L-1) and 25 (EC25) (1.2–10.1 mg.L-1), which is a good indication of high antioxidant activities. Pearson’s correlation analysis revealed that there was a moderate to very strong positive correlation (R value: 0.623–0.993) among the phytochemical properties and antioxidant activities tested. Furthermore, pigmented rice bran extracts (100–1000 ppm) showed no toxic effect against NHBL, which implies that all samples are safe for human consumption. Therefore, pigmented rice bran extracts can be used as key ingredient in the development of safe and effective functional food and pharmaceutical products.

Morpho-Anatomical Investigation on the Adventitious Rooting of Excelsa Coffee (Coffea excelsa A. Chev.) Stem Cuttings
Jeff M. Opeña, Rachel C. Sotto, Bong M. Salazar, and Calixto M. Protacio

Received: 17 December 2019/ Revised: 28 September 2020/Accepted: 10 November 2020

Excelsa coffee (Coffea excelsa A. Chev.) species is known to be hard-to-root when propagated through stem cuttings. This study sought to examine the morpho-anatomical differences between Excelsa and Robusta coffee stem cuttings in order to identify any physical hindrances to rooting and to trace the origin of adventitious rooting in Excelsa coffee. Rooting of single-node orthotropic Excelsa coffee stem cuttings from water sprouts with and without incisions in the rooting zone applied with auxin plus ferulic acid takes 5 mo and 7 mo under mist, respectively. Morpho-anatomical examinations revealed that Excelsa coffee had thicker stem structures compared to Robusta coffee. However, the most probable anatomical difference why Excelsa coffee is harder to root when compared to Robusta coffee is its narrow, compact and clustered nearly continuous layer of sclerenchyma band as opposed to the discontinuous layer in Robusta coffee. The layer of sclerenchyma physically prevented root initial development which resulted in delayed rooting in Excelsa; this was observed to occur after 4–7 mo compared to Robusta coffee where rooting occurred within 1–2 mo. Restriction of rooting was a result of the physical hindrance on root initial development by the continuous sclerenchyma band rather than by preventing root protrusion or outgrowth. Basal incisions made in the rooting zone physically disrupted the layer of sclerenchyma that enabled the development of root initials which promoted more adventitious roots in the stem cuttings.

Genetic Analyses of Abaca (Musa textilis Née) Germplasm from its Primary Center of Origin, the Philippines, Using Simple Sequence Repeat (SSR) Markers
Orlex B. Yllano, Ma. Genaleen Q. Diaz, Antonio G. Lalusin, Antonio C. Laurena, and Evelyn Mae Tecson-Mendoza

Received: 31 March 2020/ Revised: 22 November 2020/Accepted: 23 November 2020

This study presents the first extensive genetic diversity assessment of abaca (Musa textilis Née) germplasm in the Philippines using simple sequence repeat (SSR) markers. Southeast Asia is the recognized center of origin of the genus Musa, and the Philippines has the most number of abaca cultivars and wild genotypes in the world. In this study, a total of 150 accessions of abaca from Luzon, Visayas, and Mindanao, Philippines and two banana cultivars were analyzed using SSRs to examine their genetic diversity and similarity relationships. Six of 44 banana-based SSR primers were highly polymorphic, detecting 28 alleles with a mean of 4.55 alleles per locus and polymorphism information content (PIC) mean of 0.56. Genetic diversity of abaca in three main islands was high (I = 0.68). Abaca genotypes from Luzon had higher genetic diversity compared to Visayas and Mindanao. Ninety-five (95) percent was attributed to molecular variance within the population and only 5% of the molecular variance to variation among populations. Genetic analysis by Unweighted Pair Group Method with Arithmetic Mean (UPGMA) revealed nine clusters consisting of two big groups and seven small groups, irrespective of geographical origin. Using a combination of SSR markers, the abaca accessions and closely related species could be effectively distinguished and identified putative abaca duplicates. Our results provide genetic evidence of the high diversity of abaca germplasm in the Philippines. The characterization of abaca germplasm using SSR markers will aid in the identification of superior genotypes and for improving the in situ and ex situ abaca germplasm conservation and optimal utilization of abaca genetic resources.

Honey or Pineapple Juice as Extender Components for Quezon Native and Duroc Boar Semen at Different Storage Temperatures
Christian H. Castro, Geleo A. Dichoso, Marysol M. Landicho, and Percival P. Sangel

Received: 21 January 2020/ Revised: 26 August 2020/Accepted: 26 November 2020

Computer-assisted semen analyzer (CASA) was used to assess the effectiveness of formulated boar semen extenders supplemented with honey and/or pineapple juice. Four boars (i.e., Duroc = 2 and Quezon = 2) served as semen donors, where four ejaculates from each animal were used in this study. Only samples that passed the preliminary quality evaluation were further analyzed and processed. Semen samples were divided and randomly assigned into one of the 11 treatment groups: (T1) a medium-term commercial extender with antibiotics as positive control, (T2) prepared base extender without sugar and antibiotics as negative control (NC), (T3) NC + 0.5% honey, (T4) NC + 1% honey, (T5) NC + 2% honey, (T6) NC + 0.5% pineapple juice, (T7) NC + 1% pineapple juice, (T8) NC + 2% pineapple juice, (T9) NC + 0.5% mixed (1:1) honey and pineapple juice, (T10) NC + 1% mixed honey and pineapple juice, and (T11) NC + 2% mixed honey and pineapple juice and stored at either low temperature (15–20°C) or room temperature (22–25°C). Results showed that Quezon native and Duroc boar semen diluted with T1 had a semen shelf life of 48 and 52 h, respectively, which is longer compared to those diluted using other treatments. For percentage (%) slow sperm, significantly lower values were seen from using T5, T6, T7, T9, T10 and T11 than T1 for Quezon boar semen; while T1, T6 and T7 showed significantly lower values than T2 in Duroc boar semen. In terms of sperm morphological parameters, results from all treatments were comparable in Duroc boar semen; while the use of T4 had significantly lower % coiled tail sperm compared to T7 in Quezon boar semen. Room temperature was observed to be more effective in storing diluted semen from Quezon native boars; however, both low and room temperatures were comparable in maintaining diluted Duroc boar semen. Collectively, 0.5% to 2.0% of honey and/or pineapple juice, or its mixture in a 1:1 proportion, are useful and economical substitute ingredients in boar semen extender.

Evaluation of Physicochemical Characteristics, In Vitro Release and Antioxidant Properties of Inclusion Complexes Prepared with B-Carotene and B-Cyclodextrin
Jack D. Dela Cruz and Floirendo P. Flores

Received: 21 February 2020/ Revised: 10 November 2020/Accepted: 17 November 2020

The goal of this research was to prepare inclusion complexes with β-carotene and β-cyclodextrin as viable fortificants via physical blending (PB), kneading (K), and co-precipitation (CP). The physicochemical characteristics (inclusion efficiency, yield, antioxidant activities, FTIR spectra, scanning electron micrographs) besides in vitro release and antioxidant properties of the complexes were evaluated with rice as food matrix. CP and PB solids exhibited highest inclusion efficiency (83%) and yield (100%), respectively. Fourier Transform Infrared Spectroscopy (FTIR) spectra revealed changes in intensity, position, and number of peaks in β-cyclodextrin and β-carotene, confirming guest-host interactions in inclusion complexation. Scanning electron micrographs showed heterogeneous particles of β-carotene and β-cyclodextrin in PB but homogeneous solids in K and CP complexes. Simulated in vitro digestion showed burst release of β-carotene in PB complexes. Food matrix appeared to synergistically affect release and antioxidant properties. K and CP complexes exhibited sustained radical scavenging activity and ferric reducing power, respectively, and may be recommended for further applications.

Research Notes

Profiling of the Volatile Compounds of the Different Local Coffee Cultivars through Headspace – Gas Chromatography – Mass Spectrometry
Kevinilo P. Marquez and Ruel L. Mojica

Received: 01 April 2019/ Revised: 14 November 2020/Accepted: 06 November 2020

The study sought to provide the data necessary for the identification, authentication, and quality analysis of coffee products cultivated locally. Four varieties of locally cultivated coffee beans (Arabica, Robusta, Excelsa, and Liberica) were analyzed for their respective aroma profiles using Headspace – Gas Chromatography – Mass Spectrometry. For the aroma profiles, a total of 66 compounds were detected across varieties and roasting degrees based on the National Institute of Standards and Technology (NIST) Library Search Engine. An 85% mass spectral profile match percentage served as the main criteria. The aroma profiles were analyzed to differentiate the varieties and their respective roasting degrees quantitatively through the varying amounts of the volatile compounds emitted as aroma, and to determine the effects of roasting on the resulting composition of such components.

SSR Analysis of Coffea liberica var. Liberica and Coffea liberica var. Dewevrei in the Philippines
Daisy May C. Santos and Ernelea P. Cao

Received: 07 March 2019/ Revised: 02 November 2020/Accepted: 06 November 2020

In the Philippines, four coffee varieties (Arabica, Robusta, Liberica and Excelsa) are cultivated for commercial consumption. While Arabica and Robusta varieties are well-known globally, Liberica or Kapeng Barako is being developed as Philippine specialty coffee. Liberica and Excelsa are more similar in morphology compared to Arabica and Robusta since they are varieties of the same species, Coffea liberica W. Bull ex Hiern. In this study, 20 SSR markers were amplified for C. liberica specimens representing both Liberica and Excelsa varieties in the Philippines. While the SSR markers exhibited high polymorphism, between-site variation was much greater compared to between-variety. The neighbor-joining tree showed specimens from the same site clustering together with moderate to high bootstrap supports. These SSR markers may be used for geographic origin determination of coffee varieties.

Cytogenetic Analysis of Eggplant (Solanum melongena L.) and Some of Its Wild Relatives Found in the Philippines
Khris June L. Callano

Received: 14 November 2019/ Revised: 20 November 2020/Accepted: 04 December 2020

Cytogenetics is known to offer relevant information useful in understanding phylogenetic relationships, genetic mapping and plant breeding studies. In particular, the knowledge of meiotic chromosomal behavior is highly vital in working out pathways for transferring desirable traits from related species to cultivated ones. Cytogenetic characterization of Solanum aethiopicum, S. americanum, S. hirtellum, S. mammosum, S. melongena, S. pseudocapsicum and S. torvum was done using iron-acetocarmine squash technique. S. aethiopicum, S. americanum, S. hirtellum, S. melongena, S. pseudocapsicum and S. torvum were found to be diploid species with chromosome number 2n=24 while S. mammosum had a chromosome number of 2n=22. Lagging chromosomes, bridge formation and asynchronous cell division were observed in insignificant frequencies (1.96–22.06%; 12.07–20.69%; and 1.59–18.18%, respectively). Literature on the basic and applied chromosome features of Solanum species is quite insufficient in the Philippines. Results of this study can be utilized as benchmark information for future interspecific hybridization programs in eggplant.

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