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September 2020, Vol. 103 No. 3

Canna lily (Canna x generalis L.H. Bailey & E.Z. Bailey) is a genus of flowering plants native to the tropics and subtropics. More than 400 years ago, Cannas have been grown accidentally, intentionally hybridized, and traded among gardeners. As its flowers come in various blends of mixed colors and lustrous attractive foliage, Canna is one of the most popular ornamental plants used widely in landscaping settings, gardens, border plantings in parks and patios where it serves as a living fence and windbreak. Breeding and selection of novel, attractive, high-quality and climate-resilient ornamental crops are necessary to cater to the needs of the booming ornamental industry in the Philippines. To achieve such improvements and create variation, there is a need to evaluate the morphological and cytogenetic characteristics of the crop of interest such as Canna, its cultivars and putative hybrids if they are variable enough in terms of plant height, staminode size and staminode color – characteristics which are of great value to ornamental breeders and enthusiasts. {Photographs courtesy of Ms. Juareyn L. Ondoy, College of Agriculture, Sultan Kudarat State University Lutayan Campus, Sultan Kudarat, Philippines}


Physicochemical Characteristics of Soils in a Salt-affected Lowland Rice Environment and Implications for Sustained Productivity
Jocelyn D. Labios, Joy Eloiza M. Rosales and Pearl B. Sanchez

Received: 07 October 2019/ Revised: 24 July 2020/Accepted: 24 August 2020

There is lack of useful information that provides thorough understanding of the extent and severity of soil problems in salt-affected coastal lowland rice areas and their effect on farmer productivity. Soil physicochemical characterization was conducted in eight coastal rice fields in Balayan, Batangas, Philippines with varying distances from a source of saltwater intrusion. Soil, plant, and water samples were collected and analyzed from August 2014 to April 2015. Key farmer interviews were conducted. Soil organic matter was high in the wet season (WS) until the dry season (DS). Levels of soil nitrogen and phosphorus (P) were high at the end of WS and DS, indicating residuals from fertilization, and higher in fields closer to the swamp. The cation exchange capacity and base saturation in all sites were high. Zinc levels were low and may be due to high soil P. Results showed mildly alkaline soil at pH 7.4–7.7 across seasons. Soil electrical conductivity was higher in fields closer to the swamp and highest at end of WS. High exchangeable sodium percentage (ESP) indicates the presence of a high proportion of sodium in all fields regardless of distance from the swamp and at mean= 17.68% is higher than the limit set for sodic soil (15% ESP). Rice yields were within the range of varietal potential, including salinity tolerant variety Salinas, except in the field closest to the swamp which may be related to high ESP and soil sodium levels. This coastal lowland rice agroecosystem is classified as productive and the soil relatively fertile but prone to salinity and sodicity through saltwater intrusion from nearby swamps. Continued monitoring must be done so that farmers may be advised on soil fertility status and appropriate management options to sustain productivity.

Development of a Cell Suspension Protocol for Abaca (Musa textilis Nee ‘Inosa’) 
Evalour T. Aspuria, Emmanuel L. Bernardo and Maria Angie D. Tayangona

Received: 10 May 2019/ Revised: 07 August 2020/Accepted: 24 August 2020

A protocol for the establishment of embryogenic cell suspension in abaca was developed based on the International Network for the Improvement of Banana and Plantain (INIBAP) procedure for Musa. Meristematic buds from shoot cultures were excised and cultured on P5 medium for several cycles until ‘scalps’ formed. These scalps were then used to produce embryogenic complexes (ECs) in 2,4-D-containing media. Discrete primary somatic embryos (SEs) were observed on ECs 3–4 wk after initial culture. After 6 mo, out of the 150 scalps inoculated, 6% formed into ECs that had at least 10 SEs which were used to commence cell suspensions. Nine liquid media formulations were tested, and only M2 medium produced cell lines that had a characteristic bright yellow suspension indicative of embryogenic potential. However, doubling time during the initiation phase with M2 medium was lower than the doubling time in the rest of the media tested. In several months, cell lines in M2 medium stabilized with an average doubling time of about 4 wk. The old medium was replenished with fresh medium every 4–7 d, replacing at least 80% of the old medium. Maintenance of fine homogenous suspension was done at monthly intervals by transferring 2 mL aliquots with a settled cell volume (SCV) of 0.2–0.3 mL from a 1-mo-old suspension culture to an 8 mL fresh medium in 125-mL Erlenmeyer flasks. As for embryo development, regeneration media trials showed that Murashige and Skoog (MS) and M2 media promoted the formation of yellow, nodular calli, or pro-embryogenic masses (PEMs). MS and M2 medium supplemented with 1 ppm AgNO3 (with or without ascorbic acid) could enhance PEM formation, especially if the cells were pre-induced to form PEMs by gradual reduction of 2,4-D while at the cell suspension stage. Sucrose or glutamine does not seem to have any promotive effect on PEM induction during the regeneration phase. These treatments, however, were not favorable for the conversion of PEMs into mature somatic embryos. Likewise, M3 regeneration media promoted PEM formation but failed to induce somatic embryogenesis in a separate experiment. PEMs derived from the 2,4-D reduction experiment were proliferated in M2 medium and transferred in P5 for embryo development. PEMs bearing mature somatic embryos in P5 medium were transferred to abaca shoot proliferation medium where fully developed shoots were obtained in 6 mo. Although the protocol requires validation, our results clearly demonstrate that abaca could undergo in vitro somatic embryogenesis (SE) and offer a transformation platform for modern genetic improvement work.

Morphological and Cytogenetic Characterization of Four Canna (Canna x generalis L. H. Bailey & E. Z. Bailey) Cultivars and Two Putative F1 Hybrids
Juareyn L. Ondoy, Merlyn S. Mendioro, Pablito M. Magdalita, Celia B. Dela Viña and Maria Cecilia S. Reamillo

Received: 19 March 2019/ Revised: 24 July 2020/Accepted: 24 August 2020

Four Canna cultivars (C. ‘Moonbeam’, C. ‘Madeira,’ C. ‘Wintzer’s Colossal’, C. ‘Alphonse Bouvier’) and two putative F1 hybrids of C. ‘Wintzer’s Colossal’ and C. ‘Alphonse Bouvier’ (coded as H3R1 and H3R2, respectively) were evaluated for their morphological and cytogenetic characteristics. Among the morphological parameters reckoned were plant height, culm/stem diameter, flower diameter, flower color, inflorescence length, staminode length and thickness, leaf length, leaf width and leaf thickness. Acetocarmine squash technique was used to determine the chromosome number and chromosome behavior at meiosis I and II. Pollen viability was tested using I2KI test and TTZ test which was further verified through pollen germination test. Meiotic behavior was related to pollen fertility. Results showed that the four Canna cultivars and two putative F1 hybrids were morphologically variable in plant height, inflorescence length flower diameter and staminode length with measurements of 95.67–172.67 cm, 22.33–147.67 cm, 6.33–11.94 cm, and 9.95–45.87 cm, respectively. The flower color of C. ‘Moonbeam’ is Chartreuse yellow, C. ‘Madeira’ is Saturn red, and C. ‘Alphonse Bouvier’ is Vermilion while C. ‘Wintzer’s Colossal’ is Nasturtium red. The color of the two putative F1 hybrids is Chartreuse yellow for H3R1 and Jasper red for H3R2. C. ‘Moonbeam’, C. ‘Madeira’ and the two putative F1 hybrids are diploid with chromosome number 2n = 18, and basic chromosome number x = 9. Although non-congressions and laggards were observed at metaphase I and anaphase I, the chromosomes were able to catch up, hence pollen stainability and pollen germination remained high at 90.36% to 94.88%. Pollen viability for H3R1 and H3R2 were 95.75% and 90.38%, respectively. C. ‘Wintzer’s Colossal’ is a triploid with gametic chromosome number of 3n = 27, and a high frequency of irregular chromosome behavior observed during meiosis I and II. It has low pollen stainability based on results of the I2KI test while unstained pollen was observed based on the TTZ test result. C. ‘Moonbeam’ and C. ‘Madeira’ can be used as a female parent as well as a male parent due to their high pollen viability. The two putative F1 hybrids H3R1 and H3R2 had a novel staminode size and attractive colors that ornamental breeders and Canna enthusiasts may find valuable.

Isolation and Characterization of Bioactive Compounds from Seeds of Ipil-ipil (Leucaena leucocephala) and its Antifeedant Activity Against the Third Instar of Common Cutworm (Spodoptera litura)
Elmira Emery R. Medina, Annabelle T. Abrera and Marlon N. Manalo

Received: 19 October 2018/ Revised: 07 August 2020/Accepted: 24 August 2020

Bioactive compounds from ipil-ipil (Leucaena leucocephala) seeds were isolated using solvent extraction and silica gel column chromatography. Antifeedant activities of the crude ethanolic extract, chromatographic fractions and crystalline isolate were tested on third instar larvae of the common cutworm (Spodoptera litura). Dual choice assay using 10 μg cm-2 applied on both adaxial and abaxial parts of castor (Ricinus communis) leaves showed that the crystalline isolate had the highest activity, with percentage relative feeding inhibition of 79.93 ± 1.00%. The very high melting point, together with data from chemical and spectroscopic analyses, suggest that the isolate is a saponin-containing complex with molar mass of 662 g mol-1 in the major component.

Formulation of Larvicide from Capsicum frutescens (Linn.) Fruit Extract Against Aedes aegypti (L.)
Marie Claire Soria, Femina Grace Argonza, Christine Claros, Noel Quiming, Michael Russelle S. Alvarez and Bienvenido Balotro

Received: 12 February 2020/ Revised: 29 July 2020/Accepted: 24 August 2020

The crude ethanolic extract of Capsicum frutescens fruit was used in the formulation of larvicidal granules. The extract was first screened for phytochemical content and larvicidal activity against Aedes aegypti. Five trial formulations were developed using different combinations of excipients identified to be compatible with the active extract. These formulations were evaluated using Quality by Testing (QbT) Method (organoleptic evaluation, moisture content, powder flow property, particle size distribution, dissolution in water and larvicidal bioactivity). The C. frutescens extract (500 ppm) killed 100% of the test larvae after 24 and 48 h. Phytochemical analysis revealed the presence of steroids, indoles, alkaloids, flavonoids, sugars, coumarins and anthraquinones. Among the formulations tested, formulation # 5 (25% extract, 59% lactose, 1% 5% PVP-ethanol solution, and 15% microcrystalline cellulose (MCC) was the most active, with LC50-24 h=108.68 ppm, LC90-24 h=233.44 ppm, LC50-48 h=102.68 ppm, and LC90-48 h=182.50 ppm. Larvicidal granules (Formulation #5) against A. aegypti were developed. The granules were able to kill 100% of the test larvae after 24 and 48 h of exposure. In addition, the formulation passed all the quality control measurements (low moisture content, excellent powder flow, soluble in water, and 90% cumulative frequency within 0.420–0.841 mm).

Plasticity in Nodal Root Hardpan Penetration, Deep Soil Water Uptake, and Shoot Dry Matter Production under Soil Moisture Fluctuations Using Chromosome Segment Substitution Lines of Rice
Dinh Thi Ngoc Nguyen, Roel Rodriguez Suralta, Mana Kano-Nakata, Shiro Mitsuya, Stella Owusu-Nketia and Akira Yamauchi

Received: 03 February 2020/ Revised: 16 August 2020/Accepted: 24 August 2020

Rainfed lowland (RFL) fields generally experience soil moisture fluctuations (SMF) and have a hardpan layer that impedes deep rooting during episodes of drought. The development of root systems in response to SMF and the ability of roots to elongate through the hardpan when it becomes relatively soft during re-watering are key rice traits to efficiently extract water at the deep soil during subsequent periods of drought. Thus, this study aimed to identify their chromosome segment substitution lines (CSSLs) from Sasanishiki and Habataki crosses, which have root plasticity exhibited in response to SMF, which enables plants to penetrate the hardpan and develop deep root systems. Root plasticity was evaluated by comparing root traits between each of the 39 CSSLs and their recurrent parent, Sasanishiki, under water-stressed and well-watered (WW control) conditions in hydroponics and soil-filled pots. Among the 39 CSSLs, three (SL34, SL35 and SL39) had similar growth as Sasanishiki under WW, but were able to produce greater shoot dry weight than their recurrent parent under transient soil moisture stress in hydroponics and soil culture. Under SMF, in the rootbox-pinboard and rootbox-hardpan systems, only SL39 showed a significantly greater root system development than Sasanishiki. SL39 also had a more enhanced root aerenchyma formation than Sasanishiki in the shallow layer during drought-rewatered conditions under SMF, possibly facilitating atmospheric O2 diffusion to the root tips. As a consequence, SL39 promoted nodal root elongation through the hardpan during rewatering and subsequent greater deep root system development to access more water from the deep soil during the drought period of SMF, relative to Sasanishiki. The results implied that SL39 can be a good genetic material to study the QTLs associated with plasticity in root hardpan penetration and deep root system development in rice.

Post-Partum Clinicopathological and Reproductive Performance Assessment and Haptoglobin Measurement of Dairy Cattle with Retained Fetal Membrane
Masaaki Nakamura, Toru Miyamoto, Gerry Amor Camer, Takeshi Koyama, Yoshitaka Matsui, Tomochika Sugiura, Masaharu Moriyoshi, Ken Nakada and Yutaka Sawamukai

Received: 16 May 2019/ Revised: 12 February 2020/Accepted: 17 August 2020

Cattle with retained fetal membrane (RFM) interrupts optimal dairy productivity including reproductive performance. In this 28-month duration experimental study, several clinical tests (observation of morbid clinical signs, rectal temperature, vaginal discharge scoring, and blood inflammatory marker measurements) were completed in 19 cattle with RFM including 8 cattle with normally expelled fetal membrane. Twelve out of 19 cattle showed morbid clinical signs at 2.2±1.3 dpp and pyrexia at 2.3 ±1.7 dpp. All cattle with RFM presented vaginal discharge scores of 1–3 (4-point scale) that steadily persisted for 2 to 8 weeks post-partum. Lower conception rates were noted in cattle with RFM than those with no RFM (64 % vs. 100 %). Blood haptoglobin (Hp) levels of cattle with RFM and with morbid clinical signs were consistently elevated from 1–10 days (all P<0.01). The Hp of test cattle for 3 days correlated with the ‘days open’ (Rs=0.47). These findings have in effect validated the value of using Hp in assessing cattle with RFM. In addition to using Hp, regular clinical and laboratory tests as described herein should be integrated in the management of cattle with RFM.

Research Notes

In Vitro Seedling Growth of Adlay (Coix lacryma-jobi L.) `Pulot’ and Its Response to 2,4-Dichlorophenoxyacetic Acid, Benzylaminopurine, Plain Cow Dung Ash and Agnihotra Ash
Emmanuel L. Bernardo, Pamela G. Fernandez, Evalour T. Aspuria, Nina M. Cadiz and Rocky T. Marcelino

Received: 21 June 2017/ Revised: 07 August 2020/Accepted: 24 August 2020

Adlay (Coix lacryma-jobi L.) is an emerging alternative cereal crop which may benefit from plant tissue culture approaches. In this study, the morphogenic response of mature embryos of adlay to 2,4-dichlorophenoxyacetic acid (2,4-D; 1–3 ppm) and benzylaminopurine (BAP; 1–3 ppm) in Murashige and Skoog (MS) basal medium was screened in vitro. Further, agnihotra ash (AA) and cow dung ash (CDA) were evaluated as unconventional culture media additives. A sterilization procedure was first developed for mature embryos of adlay by varying sodium hypochlorite (NaOCl) concentration and the duration of surface sterilization. NaOCl at 1% (v/v, Chlorox®) using double sterilization (7 min: 8 min) was effective for surface disinfection of dehusked adlay seeds. In terms of plant growth regulators (PGRs), BAP and 2,4-D enhanced shoot growth in lower doses, but primary root growth was inhibited. Retarded shoot and root development were observed even at the lowest 2,4-D concentration and as BAP was increased. Yellow, compact callus was observed surrounding the mesocotyl and white crystalline and loose callus around the radicle region. Both types of calli appeared non-embryogenic and were most frequent at 2 and 3 ppm 2,4-D regardless of BAP concentration and ash additives. As for ash additives, application of 0.01% cow dung ash, 0.01% agnihotra ash and 1% cow dung ash resulted in larger seedlings compared with 1% agnihotra ash in basal media, although these differences were not pronounced in the plant growth regulator (PGR) background. Agnihotra ash and cow dung ash as miscellaneous additives may provide cost savings in plant tissue culture; further work is warranted.

Physio-biochemical and Antioxidative Responses of Sunflower (Helianthus annuus L.) Genotypes to Salt Stress
Md Shakhawat Hossain, Mahbub Nl Islam, Md Rezwan Mollah, Mirza Hassanuzzaman and Motari Rohman

Received: 19 August 2019/ Revised: 08 July 2020/Accepted: 23 July 2020

Salinity is one of the important abiotic stresses affecting growth, photosynthesis, ion accumulation, and antioxidant defense systems in sunflower (Helianthus annuus L.). This study investigated the physio-biochemical and antioxidative responses in different salt-sensitive sunflower genotypes: GP-4030 and BARI Surzumukhi 2 as newly identified tolerant genotypes, PS-2 as sensitive and Hysan 33 as standard tolerant. Salt stresses caused higher reduction in relative water content (RWC), chlorophyll (Chl) content, K+ and K+/Na+ ratio, photosynthetic rate (pN), transpiration rate (E), stomatal conductance (gs), and intercellular CO2 concentration rate (Ci) in salt-sensitive genotype. This genotype had higher Na+, superoxide (O2.-), hydrogen peroxide (H2O2), lipid peroxidation (MDA) and proline (Pro) content than the tolerant genotypes. The tolerant genotypes maintained higher Pro content than the salt-sensitive genotypes under salt stress. All the tolerant genotypes had higher increment of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities under salinity (12 dS m-1) than the salt-sensitive genotype. Similarly, increment of ascorbate peroxidase (APX) and glutathione reductase (GR) activity was higher in the tolerant genotypes, but glutathione peroxidase (GPX) activity was higher in salt-sensitive genotype. However, both monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) activities decreased with salinity level. Glutathione S-transferase (GST) activity increased in all the genotypes under salinity and was higher in the tolerant genotypes. Higher accumulation of Pro, along with improved physiological and biochemical parameters in the tolerant genotypes, can confer tolerance by reducing oxidative damage through up-regulating antioxidant defense under salinity.

Physicochemical and Quality Properties of Pumpkin (Cucurbita moschara Duch.) Jam, Marmalade and Fruit Leather
Sezin Seymen, Gulsah Ozcan-Sinir and Omer Utku Copur

Received: 28 May 2019/ Revised: 26 February 2020/Accepted: 17 August 2020

Pumpkin, a fruit that is rich in mineral, vitamin and fiber, is good for a healthy diet. The aim of this study was to produce jam, marmalade and fruit leather from pumpkin (Cucurbita moschata Duch.) and examine the physicochemical and quality properties of the products such as hydroxymethylfurfural (HMF), total phenolic content, total antioxidant capacity, total sugar, organic acid composition, pH, water-soluble dry matter and moisture. The highest acid content was determined in fruit leather (4.29%), followed by marmalade (0.24%) and jam (0.24%). Pumpkin marmalade (39.07 mg kg-1) contained a high level of hydroxymethylfurfural compared with fruit leather (30.96 mg kg-1) and jam (17.56 mg kg-1), while the total phenolic content of marmalade, jam and fruit leather was 71.92, 63.33 and 113.23 µg gallic acid equivalent (GAE) 100 g dw-1, respectively. Pumpkin marmalade had the highest antioxidant capacity of 13.05 μmol Trolox equivalent (TE) g dw-1.

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