Open Access Short Research Article

Effects of Cadmium Stress on Photosynthetic Physiological Characteristics of Two Rorippa Species

Yuan Yun- Ning, Wang Lin- Yu, Chen Hao, Xu Yu- Feng

Biotechnology Journal International, Page 32-42
DOI: 10.9734/bji/2020/v24i630120

The photosynthetic physiological characteristics of Rorippa amphilia and Rorippa sylvestris were studied by pot experiment under Cd stress, and 7 Cd treatments were 0, 1, 5, 10, 50, 100, 200(mg·kg-1)respectively. The results showed that, (1) The Chla, chlb, and Ch(la+b) changed little when the Cd concentration in the soil was less than 100 mg·kg-1. There was no significant difference among the treatments. When Cd concentration was more than or equal to 100 mg·kg-1, all indexes decreased significantly, Chla/b increased gradually with the increase of Cd stressful concentration; (2) Net photosynthetic rate (Pn), stomatal conductance (Gs) and transpiration rate (Tr) first increased and then decreased with the increase of Cd stressful concentration, while intercellular carbon dioxide concentration (Ci) increased significantly; (3) The photochemical quenching coefficient (qP) and electron transfer rate (ETR)  decreased gradually with the increase of Cd stressful concentration, while the non-photochemical quenching coefficient (NPQ) increased with the increase of Cd stress concentration. It indicated that the photosynthetic mechanism of leaves was damaged; the maximum photochemical quantum yield (Fv/Fm) and potential activity (Fv/Fo) of PSII decreased gradually with the increase of Cd stress concentration. When Cd concentration was more than 50 mg·kg-1, the decrease was very obvious, showing photoinhibition. The concentration of Cd below 50 mg·kg-1 had little effect on photosynthesis. These results provide a theoretical application by using these two species of Rorippa to ecological restore the Cd contaminated farmland and abandoned mines.

Open Access Short Research Article

Selection and Validation of Reference Genes for Quantitative Real-Time PCR Expression Analysis of Candidate Genes in Carposina sasakii (Lepidoptera: Carposinidae)

Zuobing Yan, Yongli Li, Zhou Zhou, Yongan Zhang, Liangjian Qu

Biotechnology Journal International, Page 75-85
DOI: 10.9734/bji/2020/v24i630124

Carposina sasakii is one of the most important pests on the quality of stone and pome fruits. Investigation of a gene expression level in the species is hampered because of the gap of validated reference genes. The expression variation in the transcription levels of eight candidate reference genes, Actin (ACT), Tubulinbeta-1 (TUB), Ribosomal protein 49 (RP49), Elongation factor1-alpha (EF-1a), Elongation factor1-b (EF-1b), Elongation factor1-d (EF-1d), Ribosomal proteinL13 (RPL13) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), were analyzed by quantitative real-time PCR (qPCR). The stability and ranking of these gene expression profiles in three organ types (head, thorax and abdomen), three developmental stages (larva, pupa and moth), and five diapause states (non-diapause, pre-diapause, diapause 0 d, diapause 20 d and diapause 60 d) were assessed using two algorithm-based methods, geNorm and NormFinder. EF-1a, ACT and GAPDH were evaluated to be the three stable reference genes based on the important observations and comprehensive analysis, whereas TUB and EF-1b showed low expression stability. Best gene combinations for different qPCR analysis in C. sasakii could be chosen from the three stable reference genes, the using of two reference genes is sufficient to effectively normalize qPCR data in C. sasakii. The study laid the foundation for gene expression analysis in C. sasakii and provided new information for the selection of reference genes.

Open Access Original Research Article

Impact of Drifts Resulting from Pesticide Application on Soil Microorganisms around Waste Receptacles in Port Harcourt City, Nigeria

D. Momoh, C. L. Eze, D. N. Ogbonna

Biotechnology Journal International, Page 1-8
DOI: 10.9734/bji/2020/v24i630118

Pesticides are toxic substance used to reduce or kill pests but the deposits on soil environment can remain there for long period of time causing adverse effects on soil microorganisms which are responsible for soil health conditions. This study was carried out to determine the impact of pesticide drifts on soil microorganisms in a waste receptacle around Port Harcourt city. Soil samples were obtained from various depths around waste receptacles with hand auger using standard analytical procedures. Microbial analysis was done according to prescribed standard methods. Characterization and identification of the isolates were based on their cultural, morphological, and cellular characteristics. Results obtained showed that the bacterial isolates were identified as Staphylococus aureus, Bacillus subtilis, Bacillus megaterium, Pseudomonas sp and Micrococcus sp while fungal isolates include Aspergillus niger, Penicillium sp, Fusarium siculi, and Aspergillus fumigatus, Aspergillus nidulas, Microsporium canis and Yeast. The results of the microbial counts revealed that Total Heterotrophic Bacteria (THB) had 2.08 ×109 cfu/g at a depth of 30-45 cm while Total Heterotrophic Fungi (THF) had 6.0×106 cfu/g before application with a mean value of 1.02×109   and    2.8 × 106 cfu/g respectively while after application THB had 4.1 × 108 and 4.6 × 108 cfu/g for 0-15 and 30-45 cm respectively while the THF recorded 1.0 × 106 and 0.6 × 106 cfu/g for 0-15 cm and 30-45 cm respectively. However there was a drastic decrease in the number of microbes in the soils after pesticide application especially at the surface soil depth of 0-15 cm. This shows that the pesticides application affected microbial population by reducing their numbers in the soil and this may in turn affect soil health and physiological status of their habitat. It is therefore recommended that proper surveillance during pesticide application should be considered to avoid drift to non-target organisms and that concentrations of pesticides to be used should be taken into account to avoid reduction in the number of microorganisms in soils because of the vital roles they play in maintaining soil health.

Open Access Original Research Article

RT-PCR Mediated Identification of SARS-CoV-2 Patients from Particular Regions of Bangladesh and the Multi-factorial Analysis Considering Their Pre and Post Infection Health Conditions

Kazi Mahmuda Akter, Tanjum Tushi, Sabrina Jahan Mily, Rashenda Aziz Mohona, Samin Anis, Adittya Kishor Chakraborty, Elmee Tabassum, Tahmid Ul Islam, Ozifatun Jannat Akhi, Israt Sultana Nishe, Badrun Nahar Laxy, Sanjida Sultana Zerin, Abdiqani Jama Roble, Md. Imran Hossain, Sharmin Ahmed, Salauddin Al Azad

Biotechnology Journal International, Page 43-56
DOI: 10.9734/bji/2020/v24i630121

Background: SARS-CoV-2 is a type of coronavirus shows bizarre features with fatality rates at 34.4%. The clinical and non-clinical features of COVID-19 might be causative factors for more rapid community spreading compared to MERS and SARS-CoV-1.

Objectives: To study the efficacy of RT-PCR based detection of SARS-CoV-2 patients. Besides, the patients' pre and post-infection health conditions depending on selective clinical and nonclinical parameters were analyzed.

Materials and Methods:  Clinical and non- clinical data were collected from 205 randomly selected COVID-19 patients, identified through RT-PCR from different areas of Bangladesh. The data were analyzed using statistical and analytical tools to illustrate the impact of the pandemic situation on the sufferers. 

Results: Molecular identification of the patients through RT-PCR has been observed as the most efficient option for detecting SARS-CoV-2 patients. Patients with the smoking habit have been reported to be more prone to the COVID-19 infection; surprisingly the non-smoker female was the worse suffers. The patients aged 60-69 years were the worst sufferers with every adverse health conditions mentioned. Professionally, 28% self-employed male and 8.19% female were infected. The most infected individuals were house wife (32.029%), as next to the health care workers (22.1%). With the increased age group, a decreased number of smokers were found.               Nebulization of 5% home and 12% hospital treated patients were provided, with Oxygen for 26% of hospitalized patients. After recovery, combined physical and psychological complications were observed in 19% of male and 11% female. The socioeconomic, environmental and geographical annotations exposed a clear relationship with the rate of infection as revealed from the nonclinical data analysis.

Conclusion: RT-PCR proved its specialty in COVID-19 detection. In addition, impacts of different clinical and nonclinical factors on the physical conditions of the nCoV patients were found significant in the research.

Open Access Original Research Article

Effects of Surface Sterilization Protocol and Haustorium Removal on Germination and Growth of Hybrid PB121 Coconut Palm (Cocos nucifera L.) Zygotic Embryos Cultured in vitro

Arnaud Agbidinoukoun, Euloge Rimson Somakpe, Serge Sètondji Houedjissin, Florent Engelmann, Corneille Ahanhanzo

Biotechnology Journal International, Page 57-64
DOI: 10.9734/bji/2020/v24i630122

Aims: This study aims to identify the best surface sterilization and evaluate the effect of haustorium suppression on in vitro germination of coconut palm (Cocos nucifera L.) zygotic embryos.

Study Design: Survival rate and contamination rate of zygotic embryos after different surface sterilization treatments, regeneration rate and organogenesis through the number of leaves and the length of shoots after haustorium suppression were determined. For data processing, the Analysis of Variance was used to compare the means which were separated according to Tukey test            (P = 0.05).

Place and Duration of Study: Coconut fruits (hybrid PB121) were collected 12 to 14 months after controlled pollination from CRAPP (Centre de Recherches Agricoles Plantes Pérennes), station of Sèmè-kpodji in Benin. Experiments were done in Central Laboratory of Plant Biotechnology and Plant Improvement, University of Abomey-Calavi and conducted from june to december in 2019.

Methodology: For the zygotic embryos surface sterilization, four treatments combining three concentrations (3%, 6% and 15%) of commercial bleach (Javel la Croix© containing 12° active chlorine) and immersion durations (5 min, 10 min and 20 min) were tested and the survival rate were determined for each treatment after two months culture. The zygotic embryos were then divided in two sets (haustorium excised embryos set and the whole embryos set) and cultured in modified Y3 medium supplemented with 7 g L-1 agar, 2.5 g L-1 activated charcoal, 5% sucrose,  6.10-3 mM 2.4 D (2.4-dichlorophonoxyacetic acid), gibberellic acid and 0.3 mM BAP(6-benzylaminopurine). After five months culture, the regeneration rate, the number of leaves and the length of shouts were recorded.

Results: The high survival rate (80%) was obtained with 6% of bleach and 20 min for the immersion duration without pre-disinfection. The suppression of haustorium have significantly increased the number of leaves (4.3 ± 0.02) and the length of shoots (16.2 ±0.7cm) compared to the whole zygotic embryos.

Conclusion: This protocol can help to ensure better surface sterilization of zygotic embryos before their in vitro culture and the development of vigorous plantlets in order to improve the slow growth of plantlets, when transferred to the greenhouse or field. 

Open Access Original Research Article

Recombinant Co-Expression of Collagen A1 (I) Fragment with the Prolyl 4-Hydroxylases (P4H) Subunits in Komagataella phaffii

Esra Avci, Zülal Kesmen

Biotechnology Journal International, Page 65-74
DOI: 10.9734/bji/2020/v24i630123

Recombinant collagen and collagen-like products are increasingly replacing animal-sourced collagen that is difficult to produce in safe and standard quality. In this study to produce hydroxylated collagen, a 400 base pair collagen fragment of the bovine COL1A1 gene was co-expressed with prolyl-4-hydroxylase subunit α (P4Hα) and prolyl-4-hydroxylase subunit β(P4Hβ) encoding the P4H enzyme in Komagataella phaffii. For this purpose, each target gene was inserted into the pPICZαA vector and then cloned in E. coli DH5α cells. Subsequently, co-expression vectors were constructed using recombinant vectors isolated from positive clones according to the in vitro multimer ligation method. All recombinant expression and co-expression vectors were transformed into K. phaffii X33 cells by electroporation. The results of reverse transcriptase-polymerase chain reaction (PCR) proved that all target genes were transcribed by recombinant strains. The expression of recombinant proteins was performed for 96 hours by methanol-fed cultivation, and the concentration of the purified proteins from the culture medium was measured by the His-Tag enzyme-linked immunosorbent assay (ELISA) method. The concentrations of rP4Hα and rP4Hβ, and rCol1 proteins expressed individually by recombinant strains were determined to be 10.69 µg/L, 10.74 µg/L, and 8.61 µg/L, respectively, while the concentrations of co-expressed rP4Hα/β and rP4Hα/β/rCol1 proteins were 7.82 µg/L and 5.02 µg/L, respectively. These results showed that the target genes were successfully expressed and co-expressed in the recombinant K. phaffii cell.

Open Access Original Research Article

In vitro Evaluation of the Fungicidal Potential of Aqueous and Methanolic Extracts of Thevetia peruviana on the Development of Rigidoporus lignosus, Causal Agent of White Root rot of Hevea brasiliensis

Serge Bertrand Mboussi, Alain Heu, Abdou Nourou Kone Nsangou, Martial Douanla Ajebe, Godswill Ntsomboh Ntsefong, Jules Patrice Ngoh Dooh, Zachée Ambang

Biotechnology Journal International, Page 1-13

Aims: The objective of this work is to evaluate the effect of (aqueous (EAq) and methanolic (ME)) extracts of Thevetia peruviana on the in vitro development of Rigidoporus lignosus.

Study Design: A synthetic fungicide (Onazol 100), two extracts at differents concentrations: C1 = 3.5 mg / ml; C2 = 7 mg / ml; C3 = 15 mg / ml; C4 = 30 mg / ml; C5 = 50 mg / ml and C6 = 100 mg / ml for EAq; C1 = 3.5 ml/ml; C2 = 7 ml/ml; C3 = 15 ml/ml; C4 = 30 ml/ml; C5 = 50 ml/ml for ME and a negative control (T = 0 mg / ml) were used.

Place and Duration of Study: The study was conducted in pathology laboratory at University of Yaounde I and IRAD of Nkolbisson in 2018.

Methodology: Daily measurements of the pathogen's development was used to evaluate the average growth (D) of the mycelium. The inhibition percentages of the different doses of EAq, ME and ONAZOL 100 were calculated after 4 to 5 days of growth of the fungus.

Results: The major result obtained compared to the rate of rot showed that the treatments EAq, ME (C6) and ONAZOL 100 were closed. The disease rates at different concentrations of C1 to C6 were 13.91 to 100% for ME, 9.34 to 100% for EAq, and 100% for ONAZOL 100.

Conclusion: The two extracts are promising, and on small and medium scale, could be an effective and cheap formulation for the control of Rigidoporus lignosus.

Open Access Review Article

Antioxidant, Hypoglycemic and Neuroprotective Activities of Extracts from Fruits Native to the Amazon Region: A Review

Klenicy Kazumy de Lima Yamaguchi, Anderson de Oliveira Souza

Biotechnology Journal International, Page 9-31
DOI: 10.9734/bji/2020/v24i630119

The Amazon forest has the largest biome on the planet, and it is estimated that only 16 to 20% of the identified animal, and plant biodiversity. Considering plant diversity, we will highlight the biological properties of the fruits extracts of Arecaceae, Caryocaraceae, Malvaceae, Myrtaceae, Sapindaceae, and Solanaceae’s families due to their significant biological actions. This review presents the antioxidant, glycemic control, and neuroprotective activities from ten fruit extracts distributed in six botanical families in the Amazon region. We obtained 801 publications (described from 2010 to 2020), of which 64 articles were selected by the benchmark previously chosen. The antioxidant effect was the dominant effect observed in the studies used for this review, followed by glycemic control and protective actions in neurons. This review provides a synopsis of the recent literature exploring the extracts from native fruits to the Amazon region that could efficiently prevent pathologies associated with oxidative stress, and cellular maintenance mechanisms. 

Open Access Review Article

Distribution, Importance and Diseases of Soybean and Common Bean: A Review

Gaurav Singh, Garima Dukariya, Anil Kumar

Biotechnology Journal International, Page 86-98
DOI: 10.9734/bji/2020/v24i630125

The crops belonging to the family Leguminosae are the second to cereal crops of agricultural significance due to their high nutritional value. Soybean is native to East Asia and was originated in China while common bean is native to Central and Southern America originated in Peru. Both soybean and common bean are erect, bushy annual herbaceous plants growing up to 150 and 40 centimeters in height, respectively. They have enormous food and medicinal values. Soybean and common bean are important leguminous crops which are excellent source of high-quality proteins, minerals, vitamins, polyunsaturated fats, and other nutrients for both human and other animal feeds. Soybean is beneficial for weight loss, diabetics as dietary supplements, bone loss in women and minimizing cancer risks while common bean is useful for preventing constipation, lowering cholesterol levels and reducing the risk of heart diseases. Pests, pathogens and weeds cause harmful effect resulting in low production of both the legume crops. The present review focuses on the overview, different importance and diseases of soybean (Glycine max) and common bean (Phaseolus vulgaris). In addition, attention has been paid on various aspects of these two legumes namely taxonomy, morphological features, origin and distribution along with their pests and diseases.