Aidar Khakimzhanov, Vladimir Kuzovlev, Aset Abaildayev*

M.A. Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan

Abstract

Chitinases are important protective enzymes in plants and belong to a special group of pathogenesis-related (PR) proteins. The main target of these enzymes is various pathogenic fungi that contain chitin in their cell walls. Under the influence of fungi, chitinases produced by plants, hydrolyze chitin, which leads to suppression of the growth and death of the pathogen. Considering their important protective role, much attention is paid to the studies of these enzymes. Individual isozymes of chitinase are often used in testing and breeding agricultural plants for resistance to fungal diseases. In this work, we studied the isoenzyme composition and a number of physicochemical properties of wheat chitinases, which are important for a better understanding of their functioning in this grain crop. It was found that in seedlings of 7 days old, chitinases are localized both inside and outside the cells. Intracellular chitinase is represented only by basic components, whereas extracellular – by acidic ones. Using an affinity sorbent, the forms of the enzyme with a chitin-binding domain (CBD) were determined, which had isoelectric points (pI) of 9.3, 9.0, 8.6, 8.2, 8.0, 7.6, 5.7 and 4.6. According to SDS-PAG electrophoresis, the molecular weight (M.w.) of these enzymes corresponded to the values of 33, 35 and 56 kDa and they were not the glycoproteins. In the spectrum of isoelectric focusing (IEF) of chitinases, exochitinases were identified, which had pI values exclusively in the acidic range of 4.3-5.2. Significant differences were revealed in the thermal stability of chitinases. The acidic components were most resistant to elevated temperatures.

Keywords: Triticum aestivum L., Seedling, Chitinase, Isoenzymes, Chitin-binding domain

Reny Herawati1*, Dwi Wahyuni Ganefianti1, Hesti Pujiwati1, Bambang Sapto Purwoko2, Iswari Saraswati Dewi3

¹Crop Production Department, Faculty of Agriculture, University of Bengkulu, Indonesia; ²Department of Agronomy and Horticulture, Faculty of Agriculture, Bogor Agricultural University,Indonesia;

³Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Indonesia.

ABSTRACT

Aluminum can possibly have direct or indirect adverse effects on plant growth; however, this effect is not the same for all plants, even in the same species.  The roots of plants are most sensitive to Al toxicity accompanied to initial symptoms such as the inhibition of cell extension and retarded development of root systems. This study was aimed to evaluate doubled-haploid (DH1) upland rice derived from anther culture to Al stress and to study the genetic diversity and population distribution of DH lines due to Al stress. Al tolerant test was carried out in a greenhouse using factorial Randomized Complete Block Design (RCBD) with three replicates. Yoshida nutrient solution containing Al of 0 and 45 ppm was the first factor, while the second was the lines obtained from previous experiments (DH1), the four parents (SGJT36, SGJT28, Fatmawati, and Way Rarem), while Dupa, and ITA131, respectively as an Al tolerant and susceptible checks.. The results showed that root length, shoot length, and shoot dry weight had high heritability values and correlated well with the observed characters. Al tolerant doubled haploid upland rice lines derived from anther culture varied widely. Based on the Relative Root Length (RRL), out of 58 lines tested, 19 genotypes were highly tolerant, 29 lines were moderate, while 10 were low. The DH1 rice derived from P3 showed high, moderate, and low tolerance, while those from P6 showed high and moderate tolerance only.

Keywords: Aluminum (Al) tolerance, Doubled Haploid (DH), Upland rice lines.

Adnan F. Al-Azzawie

 Department of Biology, College of Science, University of Tikrit, Tikrit, Iraq

Abstract

Calcium-sensing receptor (CaSR) gene polymorphism A986S (rs1801725) is a genetic factor of the calcium homeostasis and susceptibility of osteoporosis. Although, its role in premenopausal and postmenopausal women with osteoporosis is yet to be investigated. Therefore, this study was conducted to assess the CaSR gene polymorphism A986S and evaluate its correlation with biochemical parameters in premenopausal and postmenopausal Iraqi women with osteoporosis. Blood samples were obtained from 100 women (53 premenopausal and 47 postmenopausal) diagnosed with osteoporosis by specialist physicians and 70 healthy women of the same age as the control group. Serum calcium and phosphorus concentrations were estimated. Genomic DNA was extracted from the whole blood and used for polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis to detect CaSR A986S polymorphism. The results obtained showed that calcium and phosphorus levels were significantly lower (p value < 0.01) in the osteoporosis women compared with healthy groups. Frequencies of T allele and TT genotype were significantly higher (p value < 0.01) in the osteoporosis patients compared with controls, while there were no significant differences in the frequencies of the patient group. Conversely, there were no differences in the calcium and phosphorus levels and there was the presence of T allele of the CaSR A986S genotypes. Thus, our finding revealed that the CaSR polymorphism A986S was one of the genetic susceptibility factors for the premenopausal and postmenopausal in Iraqi women with osteoporosis and had little effects on mineral levels.

Keywords: CaSR gene polymorphism A986S, Osteoporosis, Iraqi Women.

Shabnam Javed¹, Amna Shoaib², Zaid Mahmood³, Saiqa Ishtiaq⁴

¹Institute of Chemistry, University of the Punjab, Quaid-e-Azam Campus, Lahore, Pakistan

²Institute of Agricultural Sciences, University of the Punjab, Quaid-e-Azam Campus, Lahore, Pakistan

³Department of Chemistry, The University of Lahore, Pakistan

⁴University College of Pharmacy, University of the Punjab, Allama Iqbal Campus, Lahore, Pakistan

Abstract

The objective of the present study to check free-radical-scavenging and hepatoprotective properties of methanolic extract of Monotheca buxifolia (Falc.) A. DC. aerial parts against isoniazid and rifampicin induced hepatotoxicity in male rats. Results exhibited that methanolic extract and all three sub-fractions (hexane, chloroform and ethyl acetate) of M. buxifolia displayed greater free-radical activity, total phenolic and flavonoid content. However, ethyl-acetate followed by chloroform sub fractions showed the better total phenolic and total flavonoid content. To assess hepatoprotective activity of methanolic extract of M. buxifolia, four groups of the rate were made. Group 1 served as a control, group 2 served as hepatotoxic group (isoniazid and rifampicin orally each 50 mg/kg), group 3 served as standard group received silymarin (200 mg/kg), and 4 served as extract treatment groups, received 500 mg/kg methanolic extract. The results suggested that level of liver markers (total bilirubin, alkaline phosphatases, alanine aminotransferases and aspartate aminotransferases) significantly elevated in group 2. The methanolic extract significantly prevented isoniazid-rifampicin by decreasing raised level of liver markers of Group 4, and also improved histopathological profile of liver. The results indicated the protective effect of methanolic extract of M. buxifolia against liver injury, which might be ascribed to presence of high level of phenolics and flavonoid along with high free-radical activity in the plant extract.

Keywords: Monotheca buxifolia, Sapotaceae, Hepatoprotective, Antioxidant, Induced liver toxicity

Shabnam Javed^1*, Amna Shoaib²,Zaid Mahmood³, Shaista Nawaz⁴,Khalid Mohammed Khan⁵

¹Institute of Chemistry, University of the Punjab, Quaid-e-Azam Campus, Lahore-54590, Pakistan.

²Institute of Agricultural Sciences, University of the Punjab, Lahore, Pakistan

²Department of Chemistry, The University of Lahore, Pakistan.

³Food and Biotechnology Research Centre (FBRC), Pakistan Council of Scientific & Industrial Research Laboratories Complex, Lahore-54600, Pakistan.

⁴H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan.

Corresponding author: shabnam.chem@pu.edu.pk, 092-323-4830505

Abstract

Monotheca buxifolia [(Falc.) A. DC.] indigenous to northern Pakistan is a broad-leaved evergreen traditional plant of pharmacological versatility. The present investigation focused on evaluating the phytochemistry, nutritional profile, analgesic activity, cytotoxicity and GC-MS profile of M. buxifolia. The result showed aerial parts of M. buxifolia had high content of carbohydrate and protein, along with sufficient amounts of macro and micronutrients. The lipophilic n-hexane fraction of M. buxifolia exhibited the presence of saponins (29.30%), flavonoids (18%) and phenols (0.46 mg GAE/100 g). Analgesic activity of lipophilic fraction was equivalent to standard drug (Aspirin) as determined by the tail flick method. The F1 (oily liquid) of lipophilic fraction showed significant cytotoxicity against lung adenocarcinoma (A549) and liver (HepG2) cell lines). GC-MS profiling identified cis-vaccenic acid followed by n-hexadecanoic acid and hexadecanoic acid, methyl ester as major bioactive compounds in the lipophilic oily sub-fraction of M. buxifolia. These results support the use of oily sub-fraction of M. buxifolia as an effective and safe candidate for treatment of cancer cell lines.

Keywords: Gurgura, Macro micronutrient, Phytoconstitutents, Tail-flicking method, MTT assay

Somchit Youpensuk^{1, 2*}, Suwimon Wanwaen¹

 ¹Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand

²Center of Excellence in Bioresources for Agriculture, Industry and Medicine, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand

Abstract

Castanopsis tribuloides and Pinus kesiya are ectomycorrhizal (ECM) host plants. They are used for reforestation in degraded forestlands in northern Thailand. Objective of this research was to compare the effects of mycelial inoculation of Russula alboareolata and Amanita princeps, edible ECM mushrooms, on C. tribuloides and P. kesiya seedlings by evaluating ECM formation, growth response and nutrient accumulation. The study period was from 2018 to 2020. Two-month old seedlings of C. tribuloides and P. kesiya were inoculated with mycelium slurries of R. alboareolata and A. princeps. The effects of the ECM fungi on the host plants were evaluated at six months after inoculation. ECM roots of C. tribuloides associated with R. alboareolata and A. princeps were irregularly pinnate, whereas ECM roots of P. kesiya inoculated with the fungi were dichotomous branching patterns. Survivals of C. tribuloides seedlings in the uninoculated, R. alboareolata and A. princeps treatments were 66.7, 76.7 and 97.2% respectively. Survivals of P. kesiya seedlings in both the uninoculated and inoculated treatments were about 95%. Inoculation with the ECM fungi significantly increased the growth and nutrient accumulations of C. tribuloides and P. kesiya seedlings. The increase in dry weight was greater for inoculated plants of C. tribuloides than for those of P. kesiya. For C. tribuloides, the increase was significantly greater for plants inoculated with R. alboareolata than for those inoculated with A. princeps. For P. kesiya, there was no difference in the increase in dry weight between plants inoculated with each of the fungal species. Seedlings of C. tribuloides and P. kesiya associated with the edible ECM fungi may be beneficial for reforestation.

Keywords: Ectomycorrhizas, Edible mushrooms, Fagaceae, Pinaceae

Yuli Siti Fatma¹, Iman Rusmana²*, Aris Tri Wahyudi², Hamim²

¹Study Program of Microbiology, Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor, Indonesia

²Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor, Indonesia

Abstract

Rice fields are a source of methane emission. Urea fertilization is considered to increase methane emission in rice field. Reduction in amount of urea applied with addition of biofertilizer, consisting of methanotrophic and N₂O-reducing bacteria, is presumably to become an innovative fertilization technique to decrease methane emission from rice field. This current work aimed to investigate the community structure of total bacteria and methane emission-related prokaryotes in rice field soil treated with urea and biofertilizer at the vegetative and generative of rice stage. Two treatments were set up in the field experiment, i.e., 100% urea (250 kg/ha) without biofertilizer (B0) and 50% urea (125 kg/ha) with biofertilizer (B1). We used Illumina-based sequencing to investigate the soil microbial community in each treatment. Results showed that the soil bacterial community had minor changes in the two treatments throughout the rice growing period. Application of 50% urea with biofertilizer (B1) did not change the dominant bacterial phyla in rice field soil, i.e., Proteobacteria. However, there were differences in bacterial composition among the two treatments. Bacterial communities were partitioned into two clusters by the treatments (B0 and B1) rather than the rice growth phase. In addition, methanogens:methanotrophs ratio in the B1 treatment was lower than that of the B0 treatment.

Keywords: Bacterial application, Metagenome, Methanotroph, Rice plants, Soil microbiom

Abdul Rehman¹, Farwa Hassan¹,Rafi Qamar^1*, Atique-ur-Rehman²

¹Department of Agronomy, College of Agriculture, University of Sargodha, Pakistan

²Department of Agronomy, Bahauddin Zakariya University, Multan, Pakistan

Abstract

Sugarcane (Saccharum officinarum L.) budchip technology is a viable alternative of conventional sett planting, where excised auxiliary buds are used as planting material. Plant growth promoters (PGPs) have ability to enhance growth of plants and thus applied to improve production. The objective of this study was to enhance the sprouting and early growth of sugarcane budchips through exogenous application of plant growth promoters and CaCl₂. It was hypothesized that application of growth regulators on budchips will improve the growth and yield of sugarcane. Keeping in view, a pot study was planned to elaborate priming effects of PGPs on sprouting and growth of sugarcane budchips, prepared by budchip scooping machine. The study was executed at Agronomic Research Area, College of Agriculture, University of Sargodha, Sargodha for two consecutive years during spring 2015 and 2016. Experiment was arranged under completely randomized design (CRD) having four treatments i.e. Control, NAA 1 mM, CaCl₂ 1 mM, NAA 1 mM + CaCl₂ 1 mM. It was observed that sugarcane budchips treated with NAA 1 mM + CaCl₂ 1 mM gave higher sprouting establishment percentage, produced taller and thicker plants, exhibited higher leaf area index and chlorophyll contents as compared to all other treatments during each year of study. Similarly, more root length, root fresh weight, root dry weight, shoot length, shoot fresh weight and shoot dry weight was achieved with application of treatment combination; NAA 1 mM + CaCl₂ 1 mM over all others including control during both years. Among all PGPs treatments, CaCl₂ 1 mM had minimum effect on all studied parameters of sugarcane but it was statistically at par with NAA 1 mM. The results of this study led to the conclusion that soaking of sugarcane budchips in NAA 1 mM + CaCl₂ 1 mM solution proved to be superior than all other PGPs treatments and control as it promotes early growth, better establishment of seedlings and improve its other characteristics.

Keywords: Sugarcane, Budchip scooping machine, Budchips, Plant growth promoters, Soaking

Frans Grovy Naibaho^1*, Adrian Hartanto², Maria Bintang³, It Jamilah², Nunuk Priyani², Ebrry Dwi Putra¹

¹Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Palangka Raya, Palangka Raya 73111, Indonesia

²Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan 20155, Indonesia

³Department of Biochemistry, Bogor Institute of Agricultural Sciences, Bogor 16880, Indonesia

Abstract

Chemical constituents of the aqueous extract of the Chinese shallot, Allium chinense G. Don. grown in North Sumatra, Indonesia was determined through gas chromatography-mass spectrometry (GC-MS). The aqueous extract was obtained from maceration in distilled water and concentrated using rotavapor (yield: 38%, w/v). The antimicrobial activity of the aqueous extract displayed broad-spectrum inhibition against Bacillus subtilis, Candida albicans, Staphylococcus aureus and Escherichia coli. The most potential bioactivity was its antifungal activity against Candida albicans with a minimum inhibitory concentration (MIC) of 62.5 µg/mL. The results also displayed a distinct composition in North Sumatran cultivar, Allium chinense,  which previously reported to be dominated by organosulfur compounds in the Chinese shallot. Furan compounds and their derivatives dominated the composition of aqueous extract. The major components identified were 5-(hydroxymethyl)-2-furancarboxaldehyde (26.65%), 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (14.64%), lactic acid (12.70%), 3,5-dihydroxy-2-methyl-5,6-dihydropyran (10.42%) and other furan derivatives (<2%) which may be responsible for its antimicrobial activity due to its furan cocktails in the extract. The major compound, 5-(hydroxymethyl)-2-furancarboxaldehyde is then revealed as a potential antioxidant based on literature reviews.

Keywords: Allium chinense, Antimicrobial, Candida albicans, Furan, Gas Chromatography Mass Spectrometry

Angela B. Austria¹, Rich Milton R. Dulay²*, Raquel C. Pambid¹

¹School of Advanced Studies, Pangasinan State University, Urdaneta City, Pangasinan, Philippines

²Bioassay Laboratory, Department of Biological Sciences, College of Science, Central Luzon State University, Science City of Muñoz, Nueva Ecija, 3120 Philippines

Abstract

Lentinus swartzii is a new record of successfully domesticated Philippine basidiomycetous mushroom. This paper highlights the chemical compositions, antioxidant and anti-diabetic properties of mycelia and fruiting body extracts of L. swartzii. The compounds present in the ethanolic extracts were determined using thin layer chromatography (TLC), and the biological properties were assessed using stable 2,2-diphenyl-1-1picrylhydrazyl (DPPH) and unstable nitric oxide scavenging activity assays and α-amylase inhibition assay. Mycelia had essential oil, triterpenes, sugars, tannins, flavonoids, fatty acids and phenols, while the fruiting body had the same except fatty acids and sugars. Mycelia ethanolic extract at 1000 µg/mL exhibited scavenging effects against DPPH (35.29%) and nitric oxide (36.04%), contained 20.25 mg gallic acid equivalent (GAE)/g sample and showed high inhibitory activity against α-amylase (81.98%). On the other hand, the fruiting body ethanolic extract at 1000 µg/mL scavenged 43.69% of DPPH and 31.75% of nitric oxide, contained 16.92 mg GAE/g sample and exhibited high inhibitory activity against α-amylase (71.08%). Therefore, L. swartzii mycelia and fruiting body could be valuable sources of bioactive compounds with antioxidant and anti-diabetic activities.

Keywords: Lentinus swartzii, mushroom biomass, anti-diabetic, antioxidant, bioactive compounds