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March 2014 – Vol. 97 No. 1

Brown rice and germinated brown rice of four Philippine rice cultivars with different amylose content, including the waxy white rice NSIC Rc13 (short and bold grains) and black rice Ominio, low-amylose white rice NSIC Rc160 (long and slender grains), and intermediate-amylose red rice Chor-chor-os.

{Photos courtesy of Mr. Henry F. Mamucod, Rice Chemistry and Food Science Division, Philippine Rice Research Institute, Maligaya, Science City of Muñoz, Nueva Ecija 3119, Philippines}

LIST OF ARTICLES

Changes in Histone H3 Modifications During Drought Response and Recovery in Jatropha curcas
Joyce A. Cartagena and Takashi Tsuge

During exposure to drought stress, the expression of certain genes that help plants to cope with stress are up-regulated, while other genes are down-regulated. Changes in the histone modification levels can contribute to the regulation of stress-inducible gene expression. In this study, the histone modifications associated with drought response in Jatropha curcas were evaluated by chromatin immunoprecipitation (ChIP) assays along the regions of drought-inducible genes such as JcERF, JcPIP2, JcBD1 and JcLEA-5. Gene activation marks histone H3 lysine 4 trimethylation (H3K4me3), histone H3 lysine 4 dimethylation (H3K4me2), histone H3 lysine 9 acetylation (H3K9ac) and histone H3 lysine 23 acetylation (H3K23ac) were investigated during and after exposure to drought stress. The results of the ChIP assays indicated that trimethylation at H3K4 and acetylation at H3K9 are activated during stress response. The results indicate that as in Arabidopsis, drought response in J. curcas also involves epigenetic mechanisms such as histone modifications on drought-inducible genes.

Selection Criteria for Improving Grain Yield of Chickpea (Cicer arietinum L.) under Gradual Water Deficit Conditions
Nahid Niari Khamssi, Kazem Ghasemi Golezani, Saeed Zehtab and Abdollah Najaphy

Water is one of the most important limiting factors in chickpea (Cicer arietinum L.) production under arid and semi-arid conditions. This research was carried out in 2007 and 2008 to evaluate the responses of three chickpea cultivars (Hashem, Arman and Pirooz) under different irrigation treatments: irrigation after 70 (I1), 70 →90 →110 →130 (I2), 70 →100 →130 (I3) and 130 (I4) mm evaporation from class A pan, respectively. The experiments were laid out in a split-plot, randomized complete block design with three replications. The irrigation treatments were assigned to the main plots and cultivars were allocated to the subplots. All traits, except leaf proline content, decreased as water deficit increased. Reductions were significant under severe water deficit treatment (I4), compared with mild water deficit (I2 and I3) and well-watered (I1) treatments. Four principal components explained 88.7% of the variation. Factor analysis showed that four main factors, namely, growth characteristics, yield and yield components, phenology and grain filling rate explained 90.5% of the total variability. Factors 1, 2, 3 and 4 explained 40.5%, 24%, 17.9% and 8.0%, respectively, of the total variation. Based on the analyses, leaf area index, number of seeds per plant, biological yield and 100-seed weight contributed mainly to the high yield of chickpea plants under gradual water deficit conditions and can therefore be used as selection criteria for improving the grain yield of chickpea.

Effects of Germination on the Proximate Composition, Antioxidant Property and Eating Quality of Brown Rice (Oryza sativa L.)
Rodel M. Bulatao and Marissa V. Romero 

Four selected white and pigmented Philippine rice cultivars with different amylose content were used to evaluate the effects of germination on the proximate composition, antioxidant property and eating quality of brown rice (BR). These include the waxy NSIC Rc13 and black rice (Ominio), the lowamylose NSIC Rc160 and the intermediate-amylose red rice (Chor-chor-os). Their cooking time and cooked rice texture before and after germination, as well as the germination capacity of BR, were likewise determined. The germination capacity of pigmented rice samples was higher than that of the white rice samples. For the proximate composition, some parameters were notably affected by germination while the others remained the same. The ash and fat content of BR samples showed significant reduction after germination except for the ash content of red rice. Only black rice had a significant increase in crude fiber while the other cultivars had a significant reduction. Germination had no effect on the protein and carbohydrate content of BR samples. Black rice samples had the highest values of antioxidant properties, except for antioxidant scavenging activity (ASA) where the highest value was obtained in red rice. Total anthocyanin and phenolic content were significantly reduced by about 23 to 40% and 21 to 28%, respectively. On the other hand, the total carotenoid and vitamin E content of all BR samples increased upon germination, though not significantly for some cultivars. Interestingly, the ASA of the pigmented rice samples was improved significantly while the opposite was observed in the white rice samples. Germinated brown rice (GBR) samples had shorter cooking time and much softer cooked rice compared with BR samples. Germination also considerably enhanced majority of the sensory properties of BR samples. The study concluded that germination is a simple way to improve the cooking and eating quality of BR with minimal effects on its nutritional and antioxidant properties.

Planting Time, Cultivar and Preservative Effects on the Growth, Yield, Postharvest Longevity and Water Relations of Gladiolus
Iftikhar Ahmad, Muhammad Abid Yasin and Ahmad S. Khan 

Effects of planting time and postharvest sucrose concentration were evaluated on five new cultivars of gladiolus (Gladiolus grandiflorus Hort., ‘Cantate’, ‘Corveira’, ‘Eminence’, ‘Essential’ and ‘Fado’). In field trials, plant growth, yield and cormel production of the cultivars were evaluated by planting corms at 15- or 16-d intervals from September 30 to November 15, 2009 (early fall to early winter) based on randomized compete block design with factorial arrangements. All cultivars performed well in terms of growth, yield and quality of flowers and cormels, when corms were planted earlier in the season (September 30) followed by October 15 compared with late planting in November. All cultivars planted September 30 or October 15 had the highest sprouting; ‘Cantate’ and ‘Fado’ planted September 30 or October 15 had the highest leaf area, while ‘Eminence’ planted September 30 had the earliest spike emergence. Late planting in November 15 also produced early spikes, but with less number of florets and poor quality. Early plantings also produced longer stems, greater floret and spike diameter, higher number of florets per spike, higher fresh and stem dry weights, and cormel number, weight and diameter, which decreased gradually with delayed planting. A second study was conducted in the laboratory to evaluate the effects of sucrose in vase solutions (0, 1%, 2%, or 4%) as a preservative, along with constant application of a biocide, 8-hydroxyquinoline sulfate (8-HQS) at 200 mg L-1, on postharvest longevity of the five cultivars. Cut stems of ‘Eminence’ and ‘Cantate’ gladioli placed in 4% sucrose plus 8-HQS at 200 mg L-1 had significantly longer vase life. Stems of ‘Eminence’ maintained relative fresh weight compared with ‘Essential’ until end of the vase life, whereas all cultivars had similar solution uptake at all sucrose concentrations. In conclusion, gladioli planted early in the season, September 30 or October 15, resulted in best growth, yield and cormel production, while 4% sucrose plus 8-HQS at 200 mg L-1 used to keep cut stems of ‘Eminence’ and ‘Cantate’ gladioli after harvest produced the longest vase life. Moreover, all the cultivars tested performed well, making them suitable to be grown for commercial cultivation in Punjab, Pakistan.

Microwave-Assisted Extraction of Phenolic Compounds from Moringa oleifera Lam. Leaves Using Response Surface Methodology as Optimization Tool
Kang Sin, Wilheliza A. Baraoidan and Pag-asa D. Gaspillo

Microwave-assisted extraction (MAE) of phenolic compounds from the processed dry leaves of Moringa oleifera Lam. was optimized by applying response surface methodology. The phenolic profile and antioxidant activities of the extract obtained under optimum conditions were also evaluated. Optimum conditions for the extraction of phenolic compounds were achieved at 5 min of extraction time, 65ºC of extraction temperature, 0.015 g mL-1 of sample/solvent ratio and 35% of aqueous ethanol concentration. The yield of total phenolic content under optimum conditions was 16.5 ± 0.2 mg gallic acid equivalent (GAE) per g of dry leaf. The extraction yield was comparable with yield obtained by the conventional extraction technique. However, the technique was time efficient. The antioxidant activities were relatively high compared with those of some plants reported in the literature. Interestingly, the presence of vanillic acid, a new compound which had not been reported in M. oleifera in prior literature, was identified from the chromatogram for the dry leaf extract. The presence of the compound might be due to the lower destructive effect of MAE, at least, against vanillic acid

Microwave-Vacuum Drying Characteristics of Carrot (Daucus carota L.)
Nazmi Izli and Sundaram Gunasekaran

Sliced carrots were dried in a constant-power microwave oven by applying microwave power in pulsed and continuous manner while subjecting the samples to different vacuum levels. The drying kinetics as well as dried product quality, such as color and rehydration ability, were determined. Experimental moisture loss data versus drying time data were fitted to nine thin-layer drying models. The Page model fit the best in describing drying kinetics when samples were dried in 10.67 kPa vacuum with pulsed application of 500 W microwave power; for all other drying conditions, the Midilli model fit was better. Carrot samples dried in the pulsed mode had higher color values (L*, a* and b*) than those dried in the continuous mode. The samples dried at 500 W continuous mode in 5.33 kPa vacuum exhibited higher rehydration ratios than the other dried samples.

Fire-Retarding Properties of Nano-Wollastonite in Solid Wood
Azar Haghighi Poshtiri, Hamid Reza Taghiyari and Ali Naghi Karimi 

Fire-retarding properties of wood boards from three woody species (Populus nigra, Fagus orientalis and Abies alba) impregnated with nano-wallostonite (NW) were studied. Specimens were prepared according to the ISO 11925 (Part III) standard. Impregnation at 4%, 6.3%, 10% and 12% was carried out using the Rueping method (empty-cell process); the treated wood samples were compared with the control specimens. Five fire-retarding properties were measured, namely, weight loss (%), ignition delay (s), flaming duration (s), glowing duration (s) and carbonization area (%). The fire-retarding properties were improved in the NW-treated specimens in all three woody species. Heat conductivity of NW limited the accumulation of heat at the spot nearest to the flame and promoted heat dissipation all over the specimens. Furthermore, NW acted as a physical barrier to deter heat and mass transfer between the gas and the condensed phase, contributing to improvement in the fire-retarding properties. NW at 4% was not enough to significantly improve the fire-retarding properties, while 12% NW was excessive and increased the treatment costs. NW at 6.3% and 10% did not show significant differences in most of the fire-retarding properties. Therefore, NW at these concentrations may be recommended for use in improving fire-retarding properties in solid wood

Photosynthetic Pigment Content, Phenol and Flavonoid Concentration, and Defense Enzyme Activity of Oriental Lily (Lilium auratum L. cv. Sorbonne) Infected with Lily Mottle Virus
Yubao Zhang, Zhongkui Xie, Hadley Randal Kutcher, Yajun Wang, Ruoyu Wang and Zhihong Guo

The effects of Lily mottle virus (LMoV) infection were determined on the photosynthetic pigment content, total phenol and flavonoid concentration, and the activities of defense enzymes including peroxidase (POD), polyphenol oxidase (PPO), superoxide dismutase (SOD) and phenylalanine ammonia lyase (PAL) of oriental lily (Lilium auratum L. cv. Sorbonne). Compared with the healthy control, the chlorophyll content varied in the LMoV-infected plants. The levels of chlorophyll a (Chl a), chlorophyll b (Chl b) and total chlorophyll (Chl a+b) in the LMoV-infected plants were either not significantly different from the control, or were significantly decreased. The effect of LMoV infection on carotenoid levels was rather variable. Moreover, total phenol and flavonoid concentration were significantly enhanced after LMoV infection, and significantly higher POD activity was observed in all the LMoV-infected plants. Significantly higher PPO, SOD and PAL activity was noted only in partly LMoV-infected plants. Furthermore, when the LMoV CP gene copies ranged between negative and 1.24×10−5, a quadratic relationship was confirmed between the values for the LMoV CP gene copies and the photosynthetic pigment content, total phenol and flavonoid concentration. Our results indicate that the reduced proliferation of viral gene copies decreased the damage of disease to some extent.

Three New Records of Defoliators (Lepidoptera: Noctuidae, Pyralidae & Sphingidae) on Some Ornamental Plants in Laguna, Philippines
Ireneo L. Lit, Jr., Merdelyn T. Caasi-Lit and Aimee Lynn A. Barrion-Dupo 

Three lepidopterous insects are reported for the first time as new defoliators of ornamental plants in Laguna, Philippines. These are the lily cutworm Spodoptera picta (Guérin-Méneville) (Noctuidae) infesting Crinum lilies, the hibiscus leafroller Haritalodes derogata (Fabricius) (Pyralidae) infesting hibiscus and related species, and the oleander hornworm Daphnis nerii (Linnaeus) (Sphingidae) infesting nerium, chichirica and related species. S. picta is also a new faunal record for the Philippines.

Potential of Fipronil as a Feeding Toxicant Against the Subterranean Termite Heterotermes indicola (Rhinotermitidae: Isoptera)
Abid Farid, Muhammad Misbahul Haq, Misbah Ullah and Abdus Sattar 

Studies were conducted to find the dose-response relationship, feeding deterrence and transfer of fipronil to unexposed nest mates when offered to the subterranean termite Heterotermes indicola in a substrate. Dose response studies revealed that values of the effective lethal time to kill 90% of the treated termites (ELT 90) ranged from 2 to 6 d for 5 to 50 ppm. Projected ELT 90 value was protracted for 1 ppm, showing a value of 11 d. Choice feeding and exposure studies indicated that no significant difference was observed between the consumption of treated and untreated substrates at a concentration range of 1– 20 ppm, while at 30 and 50 ppm, the termites preferred to feed on untreated blotting paper. When termite workers (donors) force-fed on fipronil were confined with untreated workers (recipients) for a period of 10 d, all concentrations greater than 1 ppm were able to inflict more than 50% mortality on the untreated workers. Relevance of the results is discussed with a possibility to use fipronil as a spot treatment bait in agricultural fields

Sequence Diversity of OLA-DQA2 Gene in Tibetan Sheep
X. Liu, J. Hu, Y. Z. Luo, J. Q. Wang, S. B. Li and Q. M. An

Variation in the Tibetan sheep DQA2 gene was investigated by amplification of exon 2 using polymerase chain reaction (PCR), followed by single-strand conformational polymorphism (SSCP) analysis, cloning and DNA sequencing. Seventeen nucleic acid sequences were identified. Nine alleles (OLA-DQA2*I to OLA-DQA2*Q) were newly identified in the Tibetan sheep. This latest count brings the number of alleles identified in the Tibetan sheep from 23 to 32. Phylogenetic tree analysis revealed that 2 of these 17 sequences were more closely related to cattle DQA3 or DQA4 sequences than to other sheep DQA2 sequences. These sequences clustered together and were called DQA2-like to differentiate them from other DQA2 sequences. Within the exon 2 region, 29.34% of the nucleotide positions and 56.9% of the amino acid positions were variable, and this variation seems to have arisen largely by point mutation and gene conversion. The putative antigen-binding site codons exhibited a very high rate of non-synonymous substitutions (dN/dS = 2.16), while synonymous substitutions were more frequent (dN/dS = 0.57) in the putative non-antigen binding site codons.

RESEARCH NOTES

Biomass, Sugar and Ethanol Yield, and Water Use Efficiency of Sweet Sorghum [Sorghum bicolor (L.) Moench.] Under Different Planting Patterns
Ting-Ting Xie, Pei-Xi Su, Zi-Juan Zhou, Hai-Na Zhang and Shan-Jia Li 

We evaluated the changes in biomass, sugar and ethanol yield, and water use efficiency in sweet sorghum [Sorghum bicolor (L.) Moench]. Sweet sorghum was grown in the arid region of northwestern China at three different planting patterns: one (traditional planting), two and three plants per hole. Compared with traditional planting, the aboveground dry weight, total soluble sugar, calculated ethanol yield and water use efficiency were highest in the cluster planting of three plants per hole. In addition, the total soluble sugar and ethanol yield were significantly related to stem biomass under the different planting patterns. Based on the results, the cluster planting of three plants per hole plays a significant role in enhancing yield and water use efficiency of sweet sorghum. Stem biomass had the highest contribution to ethanol yield.

Quantitative Gene Expression of C-Type Lectin in Litopenaeus vannamei Collected from Shrimp Farms with White Spot Syndrome Virus Disease Outbreak
Yasser C. Cabansag, Jonathan V. Lazaro and Apolinario V. Yambot 

White spot syndrome virus (WSSV) is one of the most devastating diseases affecting the Pacific white shrimp (Litopenaeus vannamei) worldwide. Because shrimps rely on their innate immunity to recognize and eliminate different pathogens, the gene expression of an antimicrobial peptide C-type lectin in L. vannamei collected from shrimp farms with WSSV outbreaks was quantified using real-time quantitative polymerase chain reaction (RT-qPCR). The study compared the gene expression of C-type lectin in WSSV-infected and uninfected L. vannamei. WSSV infection in shrimp from different farms was confirmed using nested polymerase chain reaction (PCR). A total of 20 WSSV-infected and 20 uninfected L. vannamei samples were subjected to quantification of C-type lectin gene expression. A significant increase in C-type lectin gene expression was manifested in WSSV-infected shrimp compared with the gene expression of uninfected L. vannamei (p<0.05). This increase indicates that the up-regulation of C-type lectin has a major role as an immune gene against WSSV infection in shrimp farm

Aeromonas hydrophila Bacteriophage UP87: An Alternative to Antibiotic Treatment for Motile Aeromonas Septicemia in Nile Tilapia (Oreochromis niloticus)
Donna May A. Dela Cruz-Papa, Charli Mae G. Candare, Gene Lorenze S. Cometa, Dominique Elloisa G. Gudez, Anna Marielle Isabella T. Guevara, Ma. Bianca Therese G. Relova and Rey Donne S. Papa

The efficiency of an Aeromonas hydrophila bacteriophage (UP87) isolated from sewage samples was compared to that of the antibiotic oxytetracycline (Liquamycin LA-200) for the treatment of “Motile Aeromonas Septicemia” (MAS) in Oreochromis niloticus. Four set-ups each containing three fishes (n = 12) were used. Nine fishes were intraperitoneally injected with a lethal dose (LD100) of 0.2 mL of A. hydrophila suspension (10-7 dilution). Two set-ups were treated with oxytetracycline or 107 plate-forming units mL-1 of UP87 phage; in the third set-up, fish remained untreated while in the fourth setup, fish remained uninfected. All fish were monitored for physical manifestations of the disease such as scale shedding, skin lesions, hyperemia and mortality. Blood was collected via venipuncture and was analyzed for the presence of A. hydrophila. The UP87 phage was able to decrease the total amount of A. hydrophila in the blood of fish and did not cause mortality among the infected fish similar to the results obtained using oxytetracycline (P value = 0.9171). The UP87 phage therefore showed potential as an alternative treatment to MAS infection in O. niloticus.