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Expression and role of defense components in Bacillus subtilis treated rice plants against Xanthomonas oryzae pv. oryzae

Toan Le Thanh1,2*, Nguyen Huy Hoang2, Kanjana Thumanu3, Channon Saengchan2, Jayasimha Rayalu Daddam4, Rungthip Sangpueak2, Narendra Kumar Papathoti2, Kumrai Buensanteai2

1Crop Protection Department, College of Agriculture, Can Tho University, Can Tho city, 94000, Vietnam

2School of Crop Production Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

3Synchrotron Light Research Institute, Nakhon Ratchasima, 30000, Thailand

4Department of Animal Science, Agriculture Research Organization, Volcani Center, Rishon Lezion 7505101, Israel


Bacterial leaf blight (BLB) of rice has a high epidemic potential and usually causes severe damage. This research was conducted to assess the efficacy and characterize the mechanism of the systemic resistance of rice plants induced by the Bacillus subtilis strain CaSUT007 to BLB. The results revealed 30% reduction in the severity of BLB in the treated rice plants, and real-time PCR measurements indicated a significant 1.1–1.2-fold increase in their concentrations of the defense genes of phenylalanine ammonia-lyase (PAL) and ascorbate peroxidase (APX). In addition, Fourier transformed infrared spectroscopy characterization of the biochemical changes in the rice leaves indicated alterations to the lignins, pectins, and amide I vibrations – these lead to the generation of defense barriers and the reinforcement of cell walls against Xanthomonas infection and invasion, thereby contributing to disease reduction. Phylogenetic trees of pal and apx revealed a significant number of polytomies among these two gene families. Moreover, analysis of the active sites of the protein PAL and APX showed one serine rotamer and a single mutation-sensitive glutamic acid residue in the region of the binding site/pocket. The possible interactions of PAL and APX with other proteins revealed insight into the defense mechanism: APX6 interacts directly with MDAR5, MDRA3, DHAR1, and other important defense proteins, while PAL has direct interactions with 4CL4, 4CLL9, and 4CL3, among other defense proteins. Therefore, treatment with the B. subtilis strain CaSUT007 promoted faster, stronger and more intense responses in rice plants against BLB.

Keywords: Bacillus subtilis, Defense genes, Leaf blight, Protein interaction

Studies on bioflocculant exopolysaccharides (EPS) produced by Anabaena sp. and its application as bioflocculant for low cost harvesting of Chlorella sp.

Amanda Putri Irawan1, Amalia Rahmawati1, Ulfa Abdila Fahmi1, Arief Budiman2, Khusnul Qonita Maghfiroh1, Tia Erfianti1, Dea Putri Andeska1, Renata Adaranyssa Egistha Putri1, Istini Nurafifah1, Brilian Ryan Sadewo2, Eko Agus Suyono1*

1Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia. Jl. Teknika Sel., Sendowo, Sinduadi, Kec. Mlati, Kabupaten Sleman, Daerah Istimewa Yogyakarta 55281, Indonesia

2Department of Chemical Engineering, Faculty of Engineering, Universitas Gadjah Mada, Indonesia. Sendowo, Sinduadi, Kec. Mlati, Kabupaten Sleman, Daerah Istimewa Yogyakarta 55284, Indonesia


Microalgae harvesting is critical to remove water from algal growth media with solid-liquid separation. Bioflocculation has the same principle as flocculation. Using solid-liquid separation, microalgae harvesting removes moisture from the algal growth substrate. The same idea underlies flocculation and bioflocculation. Using fungal and bacterial bioflocculants requires a special medium that is different from the microalgae medium, that fungi and bacteria can contaminate microalgae, so it is not recommended to be used as a bioflocculant agent. Microalgae Anabaena sp. was chosen in this study as a bioflocculant agent since it can produce exopolysaccharides (EPSs). Dissolved proteins and carbohydrates make up EPSs. This investigation looked into employing Anabaena species to extract Chlorella species. The harvest day was used to measure the parameters. A spectrophotometer was used to measure the precipitation percentages. Bligh and Dyer’s methods were used to measure lipid contents. The phenol-sulfate was used to perform carbohydrates. Bradford method was used to quantify proteins. The ratio of 1:1.25 was determined to have the best proportion of flocculation and carbohydrate content (Chlorella sp. : Anabaena sp.). The ratio of 1:1 was determined to have the maximum cell lipid and protein content (Chlorella sp.: Anabaena sp.). The application of this study will be beneficial to design effective methods for harvesting microalgae using biological materials such as other microalgae.

Keywords: Bioflocculation, Anabaena sp., Chlorella sp., Exopolysaccharides

Metabarcoding of nematode communities associated with Ananas comosus L. (Pineapple)

Leilidyn Y. Zurbano1*, Chester C. Deocaris2, Carmelita P. Mapanao3, Arnel O. Rendon3, Lourdes V. Alvarez3


1Agriculture Department, Polytechnic University of the Philippines, Lopez, Quezon Branch, Quezon Province, Philippines

2Department of Physical Science, College of Science, Polytechnic University of the Philippines, Sta. Mesa, Manila, Philippines

3Department of Biology, College of Science, Polytechnic University of the Philippines, Sta. Mesa, Manila, Philippines



Plant-parasitic nematodes are serious pests causing important crop losses worldwide. Hence, this study was conducted to determine the occurrence of plant-parasitic nematodes (PPNs) in healthy and unhealthy pineapple crops. To determine if the infestation is prevalent, a soil metabarcoding analysis was done. Six soil samples were obtained from the rhizospheres of Red Spanish pineapple farm in Mabitac, Laguna, Philippines. They were freeze-dried and brought to the laboratory for metagenomics analysis using Primers NemF and 18Sr2b. The results showed uncultured Eukaryotes (43.1%) nematodes (31.2%), Platyhelminthes (6.9%), Apicomplexa (6.5%), Annelida (5.0%), Rotifera (3.6%), Arthropoda (1.8%), Ascomycota (1%), unidentified (0.4%) and Basillariophyta, Cercozoa, Chytridiomycota, Mollusca and Mucoromycota with 0.1%. In total, 374,410 sequence reads were obtained and were clustered into 117 OTUs at 97% similarity. In assessing the nematode community structure, it yielded 26,565 nematodes; 5,315 nematodes were obtained from the rhizosphere of healthy samples, and 21,250 were from the unhealthy ones. The fungivore Apelenchus avenae (Bastian 1865) was the most prevalent (47.8% and 43.9%) in both locations. Other nematodes found on the plant rhizospheres were Rhabdolaimus aquaticus (de Man), Acrobeloides varius, Rotylenchulus reniformis (Linford and Oliveira, 1940), Aphelenchoidinae sp., Panagrolaimoidea sp., and two unidentified nematodes. The Basiria sp., Alaimus sp., and two other unidentified nematodes were the only ones found in the rhizosphere of unhealthy pineapple plants. Meanwhile, Mesocriconema onoense, Aphelenchoides sp., Ditylenchus gilanicus, and an unidentified nematode were found in the rhizosphere of healthy pineapple plants. Phylogenetic analyses of the nematode communities show that only Orders Rhabditida and Enoplida are associated with the crop and the Family Aphelenchoididae is distant from Rhabditida, thus, dividing the clade into three. The first clade consists of diverse nematode communities of fungivores, bacterivores, predators, and plant and insect parasites. The second clade consists of bacterivore nematodes found in moist environments, and the third clade comprises mainly Aphelenchoididae species, mostly plant parasites.


Keywords: Metabarcoding, Acrobeloides varius, Aphelenchus avenae, Ditylenchus gilanicus, Mesocriconema onoense, Rhabdolaimus aquaticus, Rotylenchulus reniformis

Seed treatment with 24-epibrassinolide improves wheat germination under salinity stress

Otie Victoria1,2, Udo Idorenyin3, Matsuura Asana4, Liu Jia2, Liang Shuoshuo2, Shao Yang5, Itam Michael Okoi6, An Ping2*, Eneji Anthony Egrinya1

1Department of Soil Science, Faculty of Agriculture, Forestry and Wildlife Resources Management, University of Calabar, P.M.B. 1115, Calabar, Nigeria

2Arid Land Research Centre, Tottori University, Hamasaka 680-0001, Tottori, Japan

3Department of Crop Science, Faculty of Agriculture, Forestry and Wildlife Resources Management, University of Calabar, P.M.B. 1115, Calabar, Nigeria

4Faculty of Agriculture, Shinshu University, 8304, Kamiina County, Nagano 399-4598, Japan

5College of Plant Science & Technology, Huazhong Agricultural University, Wuhan 430070, China

6Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing 48824, MI, USA



Salt stress is a key ecological challenge to wheat establishment at the early stage of germination, especially in drylands. A germination experiment was conducted to determine whether an exogenous seed treatment with 24-epibrassinolide could mitigate salinity stress effects on wheat germination. Seeds of the Sudanese wheat cv. Imam were treated with 24-epibrassinolide (BR1) or without (BR0) at eleven concentrations of sodium chloride (NaCl) (0.00, 1.56, 3.13, 4.69, 6.25, 7.81, 9.38, 10.94, 12.50, 14.06 and 15.63 dSm-1), in a 2 x 11 factorial experiment arranged into a completely randomized design. Seed germination was progressively delayed with increasing salinity and the daily germination was reduced significantly. The germination average time and relative injury rate increased considerably (p≤0.05) at salt levels of 7.81 dSm-1or more. The inhibitory effects of salinity on germination were significantly (p≤0.05) reversed by seed treatment with BR1. Wheat cv. Imam tolerated salt stress up to 6.25 dSm-1 at BR0 with respect to velocity of germination, germination rate, final germination rate, germination percentage and germination index, relative to no salt (0.00 dSm-1). An early uniform establishment of wheat in saline media could be enhanced by seed treatment with BR.


Keywords: Arid region, Plant growth hormone, Triticum aestivum (L), Abiotic stress

Abundance of arbuscular mycorrhizal fungi in the rhizosphere of healthy and declining citrus in East Nusa Tenggara, Indonesia

Lily Ishaq*, Agnes V Simamora, Peters O. Bako, Yoke I Benggu, Moresi M. Airthur, Effy Roefaida, Ellias St O Nguru


Department of Agrotechnology, Faculty of Agriculture, Nusa Cendana University. Jl. Adi Sucipto Penfui Kupang, East Nusa Tenggara, Indonesia



Citrus is an important commercial crop in Timor Tengah Selatan, East Nusa Tenggara, Indonesia, but many trees are suffering a decline in health. As citrus is heavily dependent on arbuscular mycorrhizal fungi (AMF), this study investigated the abundance of AMF in the rhizosphere of healthy and declining trees in citrus orchards at 12 geographical locations. In each orchard, 6 soil and 6 root samples representing 3 healthy and 3 declining trees were collected. The soil was analyzed for AMF spore abundance as well as physical (texture) and chemical properties (organic carbon, nitrogen, phosphorus, potassium, pH, and soil exchangeable capacity), while the fine roots of citrus were assessed for colonization. The data were analyzed using analysis of variance (ANOVA) where the health condition of the trees was under the geographical location/site factor. The results showed that the abundance of AMF spores was significantly affected by the geographical location from where the soils were collected, but the health condition of the trees had no effect. However, AMF colonization was significantly affected by both site and tree health. The number of AMF morphotypes tended to be higher under healthy trees than under declining trees. Soil analysis indicated that soil fertility (N and organic C) may be important for tree health. These results provided a new perspective on the possible involvement of AMF and soil nutrients in citrus decline. Further studies are required to define the interactions between AMF, soil fertility, and disease incidence to identify strategies for managing citrus decline in the region.


Keywords: Arbuscular mycorrhizal fungi, Citrus decline, Orchard management, Orchard location, Southeast Asia




Establishment of rice yield prediction model using soil compaction

Van Huu Bui, Huu Cuong Nguyen, Quang Hieu Ngo*

Department of Mechanical Engineering, Can Tho University, Vietnam


Soil compaction has a real effect on rice yield in the Mekong Delta. Two field experiments were carried out during 2019 Summer-Autumn and 2020 Summer-Autumn in An Giang Province (Mekong Delta). OM18 rice was cultivated in the plots which were laid out in a randomized complete block design measuring 0.5 × 0.5 m with 5 and 6 m alley between blocks and between plots. The Pearson’s correlation test was applied to compare the mean and standard deviation of the soil layers and evaluate the correlation between soil compaction and rice yield in both crops. The present research results showed that the value of soil compaction increased with depth and differed among locations in the rice field. Soil compaction at 10 cm from the surface had a positive correlation with rice yield. Therefore, the prediction model of rice yield is able to build up due to soil compaction at 10 cm from the surface. Moreover, this study provides that the value of 10 cm soil layer compaction ranging between 165 and 190 kPa can be the optimal value of soil tillage for paddy rice cultivation with the highest yield in the Summer-Autumn crop.

Keywords: Soil depth, Soil compaction, OM18 rice, Rice yield, Prediction model

Occurrence of Shiga toxin producing E. coli in zoo animals of Rawalpindi and Islamabad zoos

Muhammad Basit Rasheed1, Aitezaz Ahsan2, Hamid Irshad2*, Muhammad Armaghan Shahzad2, Muhammad Usman2, Aayesha Riaz3, Tamoor Hamid Chaudhry4, Afreenish Amir4, Mohsina Zubair5, Asghar Khan6, Arfan Yousaf6

1Jungle World Theme Park and Zoo, Army Heritage Foundation, Ayub National Park, Rawalpindi, Pakistan

2Animal Health Research Laboratories, Animal Sciences Institute, National Agricultural Research Centre (NARC), Islamabad, Pakistan

3Department of Pathobiology, Faculty of Veterinary and Animal Sciences, PMAS-Arid Agriculture University, Rawalpindi, Pakistan

4Public Health Laboratories Division, National Institute of Health, Islamabad, Pakistan

5National Environmental Quality Standard, Pakistan Environmental Protection Agency, Islamabad, Pakistan

6Department of Clinical Studies, Faculty of Veterinary and Animal Sciences, PMAS-Arid Agriculture University, Rawalpindi, Pakistan


Shiga toxin-producing E. coli (STEC) are considered pathogens of zoonotic importance. Zoo animals have been reported as reservoirs of STEC and many STEC human outbreaks have been linked with zoo animals. Available information about the occurrence of STEC in zoo animals in Pakistan is limited. Therefore, the current study was executed to estimate the occurrence of STEC in zoo animals of two zoos of Rawalpindi and Islamabad cities in Pakistan. Total of 110 faecal samples were collected from 24 species of zoo animals. The samples were analysed for determination of eae, stx1, stx2, and ehxA genes using multiplex PCR. The positive samples for any of these genes were further analysed for isolation using sorbitol MacConkey agar. Out of 110 fecal samples, 15 samples (13.6%) contained targeted virulence genes (stx1, stx2, eae, ehxA). Six different combinations of virulence genes were observed in positive samples. Only two E. coli isolates with targeted virulence genes could be isolated from PCR positive samples. The study indicated that the wild animals maintained in zoos of Rawalpindi and Islamabad are carriers of STEC and may be the source of infection for humans.

Keywords: STEC, Wild animals, Zoo, Virulence genes, Pakistan

Performance of broiler birds on feeding natural anti stressors in summer during heat stress

Ali Salman Ajmal1, Zahid Hussain2*, Muhammad Moazam Jalees3, Jamila Shafi4, Sohail Manzoor5, Anwar ul Haq6

1Department of Poultry Production, University of Veterinary and Animal Sciences, Lahore, Pakistan

2Livestock & Dairy Development Department, Lahore, Pakistan

3Department of Microbiology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur, Pakistan

4Poultry Disease Laboratory, L&DD, Samundri, Pakistan

5Animal Disease Diagnostic, Reporting and Surveillance, L&DD, Lahore, Pakistan

6Veterinary Research Institute, L&DD, Lahore, Pakistan


This study was planned to assess the effectiveness of natural anti heat stressors on the broilers during hot climate. Three hundred one day old broiler birds were divided into four treatments groups, yoghurt, garlic powder and mint and a control group. After two weeks of rearing, data were recorded for growth performance, carcass and visceral organ yield and immune response against Newcastle Disease. Statistical analysis revealed that growth performance (weight gain, carcass weight and cumulative feed conversion ratio (FCR) was significantly improved with garlic powder, but feed intake remained non-significant. Dressing% and breast% were also significantly higher in garlic fed birds and similar effect was observed for intestinal length. Among visceral organs, liver% was improved upon garlic supplemented broiler but heart and gizzard were not influenced by dietary treatments. Additionally, garlic powder in feed also resulted in significantly higher antibody titer against Newcastle disease. It was concluded that garlic as natural anti-heat stressor has significant positive influence on the performance of heat stressed broilers.

Keywords: Broiler birds, Heat stress, Garlic, Mint, Yogurt

Rose and eucalyptus essential oil as potent anti-liver cancer agents

Shabnam Javed1, Amna Shoaib2, Ayesha Malik3, Bushra Ijaz3, Shagufta Perveen2 

1Department of Organic Chemistry, School of Chemistry, University of the Punjab, Quaid-e-Azam Campus, Lahore-54590, Pakistan

2Department of Plant Pathology, Faculty of Agricultural Sciences, University of the Punjab, Lahore, Pakistan, Quaid-e-Azam Campus, Lahore-54590, Pakistan

3National Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore-53700, Pakistan


The present study was designed to investigate the anti-cancer potential of essential oil obtained from Rosa indica (REO) and Eucalyptus citriodora (EEO) against the liver carcinoma cell line (HepG2). Firstly, the cytotoxic activity was assessed using increasing concentrations ranging from 3.12 to 200 µg/ml via MTT assay. EEO showed only 2% cell viability while REO represented 18% at the highest concentration (200 μg/ml). The half-maximal inhibitory concentration (IC50) of EEO and REO was found to be 17.741 µg/ml and 18.55 µg/ml respectively. Additionally, evident morphological changes in HepG2 cells were observed after 24 hours of essential oil treatment compared to control or untreated cells. Furthermore, to strengthen the anti-cancer perspective of essential oils, the anti-metastatic potential was evaluated through the wound healing assay. EEO promisingly inhibited migration (4% wound closure, **p > 0.01) in HepG2 cells after 24 hr treatment. Likewise, REO also exhibited good results (37% wound closure, ***p > 0.001). Conclusively, the present investigation provides preliminary results which suggest that REO and EEO are potent anti-cancer agents against hepatocellular carcinoma.

Keywords: Essential oil, Metastasis, HepG2, Cytotoxicity, Liver cancer

Effect of salinity stress on physiological aspects of pumpkin (Cucurbita moschata Duchesne. ‘Laikaotok’) under hydroponic condition

Worasitikulya Taratima1*, Narisa Kunpratum2, Pitakpong Maneerattanarungroj3

1Salt Tolerance Rice Research Group, Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand
2Department of Biology, Faculty of Science, Naresuan University, Pitsanulok 65000, Thailand
3Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand


Pumpkin (Cucurbita moschata) ‘Laikaotok’ is an important edible vegetable but growth and crop yield are affected by salinity stress. Response of physiological traits to salinity stress was investigated under hydroponic culture using Hoagland’s solution at different NaCl concentrations of 0, 15, 30, 45, 60 and 120 mM. All treatments were cultured for four weeks and physiological traits were recorded. Results showed that pumpkin growth decreased after NaCl treatment, especially at 45 mM and higher concentrations. Leaf number, leaf width, leaf length, root number, stem length, stem diameter, SPAD unit, chlorophyll fluorescence in dark condition for 30 min (Fv’/Fm’), chlorophyll fluorescence in light condition (Fv/Fm), total chlorophyll, chlorophyll a and chlorophyll b contents significantly decreased (p < 0.05). Results indicated that salt concentration at less than 45 mM NaCl was suitable for C. moschata ‘Laikaotok’ growth. Phenotypic correlation coefficient estimation of 12 physiological traits showed positive correlation at a highly significant level, except for the correlation between plant height – Fv/Fm; plant height – Fv’/Fm’; root number – Fv/Fm and root number – Fv’/Fm’. This is the first report on salt stress response in C. moschata ‘Laikaotok’. Knowledge gained will be useful for pumpkin culture or rootstock selection under sensitive and moderately sensitive saline conditions.

Keywords: Growth, Hydroponic culture, NaCl, Pumpkin, Phenotypic correlation