Category Archives: b_original_articles

Original Articles

Transcriptome analysis of flower development and mining of genes related to the flower development in Oncidium

Nengyan Fang, Yuanhua Luo, Ronghui Fan, Xiuxian Ye, Minling Huang*, Huaiqin Zhong*

Institute of Crop Sciences, Fujian Academy of Agricultural Sciences (FAAS), Fujian Engineering Research Center of Characteristic Floriculture, Fuzhou, 350013 China

Abstract

Oncidium, a kind of orchid plants characterized with unique flower, is one of the four tropical orchids with high ornamental value and favored by consumers. However, our understanding about the molecular basis of its flower development is still limited. Here, we collected Oncidium tissues at different developmental stages for RNA-seq. A total of more than 621 million clean reads were generated. 134,640 unigenes were assembled, and 54,221 unigenes were annotated. The number of DEGs (differentially expressed genes) was the largest in the comparison group F2-vs-F3 (F2 stage compared with F3 stage in flowers). The GO (Gene Ontology) terms and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways enriched for the specific DEGs were diverse at different stages. The common pathways enriched in multiple comparisons were also obtained. 11 key genes were obtained in cutin, suberin and wax biosynthesis pathway. Moreover, 32 candidate genes related to flower development were screened. Most of them presented tissue-specific expressions, especially MADS-box genes described by the ABCDE model. In all, the present data provides a valuable resource for dissecting the molecular mechanism of Oncidium in regulating flower development.

 Keywords: Candidate genes, DEGs, Flower development, MADS-box genes, Oncidium, RNA-seq

Sorting effects of X/Y sperm on in vitro fertilization of Belgian blue bulls

Peng Niu1, Fei Huang1, Xin Liu2, Bo Liu2, Jie Wang1, Di Fang1, Ahrar Khan4,5*, Qinghua Gao1,2,3*

1College of Life Sciences and Technology, Tarim University, Alar, Xinjiang 843300, China

2College of Animal Science and Technology, Tarim University, Alar, Xinjiang 843300, China

3Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production & Construction Corps, Alar, Xinjiang 843300, China

4Shandong Vocational Animal Science and Veterinary College, Weifang, China

5Faculty of Veterinary Science, University of Agriculture, Faisalabad-38040, Pakistan

Abstract

There is a large market in China for the production of Belgian blue bulls, which would benefit from semen flow sorting for animal sex control. The sperms of the Belgian blue bulls were separated by flow cytometry, and then the quality of sorted sperm was tested by in vitro fertilization (IVF) and artificial insemination (AI). The fresh semen of 8 Belgian blue bulls was individually collected and processed for sorting. Sperm sorting was carried out using MOFLO cell sorter and cryopreservation. There were no significant differences in sperm motility, acrosome integrity, and DNA integrity between sex-sorted and non-sorted sperm (p > 0.05). The purified sperm with higher viability was fertilized with mature oocytes in vitro, co-cultured to cleavage and blastocyst stage, embryo sex was identified by nested PCR amplification of AMEL gene, and there was no significant difference between sorted sperm and non-sorted sperm (p > 0.05). For artificial insemination, the pregnancy rate using non-sorted sperm was higher than for sorted sperm (p < 0.05). After delivery, the sex ratio of offspring using X- and Y-sperm was 100% and 90.9%, respectively, with a significant deviation from conventional semen (p < 0.05). The birth weight of male and female calves in the control group was similar to that in the sex-sorted sperm (p > 0.05). In summary, after artificial insemination using sex-sorted sperm, normal Belgian blue calves with the predicted gender can be produced. It is of great significance to improve the commercial promotion and production efficiency of semen after sorting Belgian blue bulls.

Keywords: Belgian blue bull, Flow cytometry, In vitro fertilization, Artificial insemination, Sex ratio

Immune enhancing activity of garlic polysaccharide, selenizing Codonopsis pilosula compounds in chickens

Li Liang-liang1,2, Liu Kuan-hui3, Xie Shi-hui3, Li Ting3, Zhu Guang-huang3, Mikhlid H. Almutairi4, Muhammad Zahid5, Mushtaq Ahmad Gondal6, Qudratullah6, Khalid Mehmood5*, Yi Wu1*

1College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China

2College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China

3Wuhu Institute of Technology, Wuhu, China

4Zoology Department, College of Science, King Saud University, Riyadh, Saudi Arabia

5Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan

6Cholistan University of Veterinary and Animal Sciences Bahawalpur, Pakistan

Abstract

Codonopsis pilosula polysaccharide is a main ingredient of Codonopsis. The key component in Condoposis is identified as Codonopsis pilosula. This Codonopsis has multiple functions including stimulation of spleen, lungs and tonifying the middle Jiao and Qi. Allium sativum has functions of regulating vital energy, appetizing, detoxification, and disinfection. Polysaccharide of the bulbous is key component to perform the functions of this bulbous. In current study, the immune enhancing activity of GP, sCPP and GP-sCPP through in-vivo and in-vitro trials was evaluated. For in-vivo trials, a total of 150 one day old white roman chicks were randomly divided into five groups, namely immune control (VC), blank control (BC), GP-sCPP, sCPP and GP groups. Chicks in group GP-sCPP, sCPP and GP was intramuscularly treated with 0.5 mg, 0.5 mg and 4.5 mg of polysaccharide, respectively, in all experimental days, and chicks in VC and BC were treated with equal volume of normal saline injections daily. All chicks were bred according to the standards for the rearing of laying hens. For in-vitro trials, lymphocyte cells were cultured in a 12-well plate along with different concentrations of GP-sCPP, sCPP and GP were supplemented in the cells and incubated at 37.5℃. After that, MTT was employed to measure the cytotoxicity of those polysaccharides. The results showed that GP, sCPP and GP-sCPP could increase immune-enhancing activities by releasing cytokines of IL-2 and IFN-γ. In-vivo experiments in chicks showed significantly higher serum antibody titer, T lymphocyte proliferation, and IFN-γ, IL-2 and IL-6 levels in GP-sCPP group. These results demonstrated that GP-sCPP can significantly promote immune enhancing activity, which could be recognized as a novel candidate for new type of immune modulator.                          

Keywords: Lymphocyte proliferation, Codonopsis Pilosula, Chickens, mRNA expression, Antibody titer, Cytokines

Sciadopitysin attenuates paraquat induced renal toxicity by modulating Nrf-2/Keap-1 pathway in male albino rats

Muhammad Umar Ijaz1*, Ayesha Kalsoom1, Ali Hamza1, Nazia Ehsan1

1Department of Zoology, Wildlife and Fisheries, University of Agriculture, Faisalabad, Pakistan

Abstract

Paraquat (PQ) is a herbicide that has the potential to instigate nephrotoxicity in animals and human. Sciadopitysin (SPS) is a biflavonoid that is extracted from Taxus cuspidate and displays diverse biological activities including anti-oxidant, anti-inflammatory and anti-apoptotic. Therefore, the present investigation was designed to evaluate the mitigative potential of SPS against PQ prompted renal toxicity in albino rats. 48 male albino rats were divided into 4 groups, such as control group, PQ treated group (5 mgkg-1), PQ + SPS co-treated group (5 mgkg-1 and 2 mgkg-1 respectively) and only SPS treated group (2 mgkg-1). The exposure of PQ significantly reduced Nrf-2 as well as anti-oxidant enzymes expression, while increasing Keap-1 expression. Moreover, anti-oxidant enzymes such as, glutathione reductase (GSR), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione-S-transferase (GST), heme oxygenase-1 (HO-1) and glutathione (GSH) activities were decreased. However, in PQ-treated rats malondialdehyde (MDA) and reactive oxygen species (ROS) contents were significantly increased. PQ exposure also increased the serum level of urea, urinary protein, urobilinogen and creatinine while decreased creatinine clearance and albumin protein levels. Moreover, KIM-1 and NGAL levels were also increased in PQ exposed rats. Additionally, inflammatory indices including nuclear factor kappa-B (NF-κB), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) interleukin-1β (IL-1β) and cyclooxygenase-2 (COX-2) activity were increased in PQ administrated rats. Besides, it escalated the Bax and Caspase-3 expression. Contrarily, a substantial decrease was observed in anti-apoptotic marker, Bcl-2 expression. The exposure of PQ also induced significant histopathological damages in renal tissues. Nevertheless, SPS supplementation recovered all these damages due to its anti-apoptotic, anti-oxidant and anti-inflammatory nature.

Keywords: Paraquat, Sciadopitysin, Renal damage, Oxidative stress, Inflammation

The effects of Cryptosporidium infection on gut fungi and enzyme abundance in Sus domesticus

Wang Lei1,3, Lu Sijia1, Zou Wen1, Sun Na2, Muhammad Usman Saleem5, Mudassar Nazar6, Mohammad Farooque Hassan7, Farid Shokry Ataya8, Kong Qinghui4, Li Kun1*

1Institute of Traditional Chinese Veterinary Medicine & MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China

2Shanxi key lab. for modernization of TCVM, College of Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi, China

3College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China

4Key Laboratory of Clinical Veterinary Medicine in Tibet, Tibet Agriculture and Animal Husbandry College, Linzhi, Tibet, China

5Department of Biosciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan

6University of Agriculture Faisalabad, Sub-Campus Burewala 61010, Pakistan

7Shaheed Benazir Bhutto University of Veterinary & Animal Sciences, Sakrand, Sindh, Pakistan

8Department of Biochemistry, College of Science, King Saud University, Riyadh 11451, Saudi Arabia

Abstract

Pigs are known to be natural hosts of Cryptosporidiosis, which poses a serious threat to public health. According to the World Health Organization (WHO), Cryptosporidiosis is the major cause of severe diarrhea which causing death in infants. Many researchers suggest that gut fungi have an indispensable function in host metabolism and immunity. However, few studies have been performed to understand how Cryptosporidium infection induces alterations in the intestinal fungal communities of Tibetan pigs. Therefore, fecal samples from Cryptosporidium infected and Healthy Tibetan pigs were examined by internal transcribed spacing (ITS) gene amplification sequencing. Results showed that a total of 2 phyla, 5 classes, 9 orders, 10 family, 13 genera of fungi were detected from both the H (Healthy Tibetan pigs) and INF (Cryptosporidium infection pigs) groups. The results manifested that proportion and profile of the total fungal population obviously changed under the Cryptosporidium infection, marked by a reduction in the abundance of beneficial fungi, i.e., Leotiomycetes (p < 0.05), Aspergillaceae (p < 0.05), Penicillium (p < 0.05), Xenoacremonium (p < 0.05) and Eurotiales (p < 0.05) and an increase in the abundance of disease-causing fungi that threaten health, i.e. Colletotrichum (p < 0.05) and Clarireedia (p < 0.05). In addition to these changes, some enzymes including Arabinose-5-phosphate isomerase, Quercetin 3-0-methyltransferase and Amylosucrase were found to be significantly altered (p < 0.01) after Cryptosporidium infection using fungal function prediction analysis. This study also focused on the Cryptosporidium infection in Tibetan pigs in terms of gut fungal diversity, composition, and the abundance of enzyme. It is providing a better understanding of Cryptosporidium infection in Tibetan pigs and insights for further development of therapeutics against Cryptosporidium from the gut fungal perspective.

Keywords: Sus domesticus, Tibetan pig, Cryptosporidium, Gut fungi, Enzyme, Microbiota

Biological activity of leaves of three Morinda species detected by stimulation and suppression of gene expression of collagen, elastin, melanin and other related genes

Runglawan Sudmoon1, Sanit Kaewdaungdee2, Tawatchai Tanee3, Pornnarong Siripiyasing4, Arnold Ang5, LiTing Lee5, Xue Jing Wong5, Shiou Yih Lee5, Arunrat Chaveerach2*

1Faculty of Law, Khon Kaen University, Thailand

2Department of Biology, Faculty of Science, Khon Kaen University, Thailand

3Faculty of Environment and Resource Studies, Mahasarakham University, Thailand

4Faculty of Science and Technology, Rajabhat Mahasarakham University, Thailand

5Faculty of Health and Life Sciences, INTI International University, Malaysia

Abstract

Morinda coreia, M. elliptica and M. tomentosa leaves were investigated. α-EG and pinoresinol were found by GC-MS and GC-FID. MTT and comet assays did not reveal toxicity in normal PBMCs and DHFa cells, but the ethanol and hexane M. coreia and M. tomentosa extracts were toxic to CHL-1 cancer cells. Biological activities were detected by gene expression through qRT-PCR. Ethanol M. tomentosa extract can stimulate collagen type I, II, III and elastin creation through expression of COL1A, COL1A2, COL2A1, COL-II, COLL2, COL3A and ELN genes; inhibit enzyme synthesis for collagen and elastin degradation through ELANE, MMP1, MMP13, TIMP1 genes; does not affect DHFa cells, but does affect CHL-1 cells through the NRF2 gene; can stimulate DHFa cell proliferation through the FGF1 and FGF7 genes; has anti-inflammatory effects on DHFa cells, and has no effects on CHL-1 cells through the TGF-β gene. Ethanol M. tomentosa extract can suppress melanin production through the α-MSH and TYR genes. In summary, M. tomentosa can be used in human health and skin care products, the research was accomplished with the aim of deducing how more plant species with high bioactivities can be utilized without toxicity, the leaves of M. tomentosa can be easier to use correctly than fruits depending on the season and smell.

Keywords: Collagen, Elastin, α-EG, Morinda species, Pinoresinol, qRT-PCR

Optimization and antimicrobial properties of biosurfactant production by four indigenous soil bacterial species

Oghenerobor B. Akpor1*, Elisha O. Ezekudo1, Oguntope A. Sobajo1, Paul A. Edoh2, Samson O. Mabayoje1

1Department of Biological Sciences, Afe Babalola University, Ado-Ekiti, Ekiti State, Nigeria

2Department of Civil Engineering, Afe Babalola University, Ado-Ekiti, Ekiti State, Nigeria

Abstract

Biosurfactants of microbial origin are metabolites; hence their production is dependent on the growth of the producing microbe. The objective of this study was to assess the optimum conditions for biosurfactant by four bacterial species. Biosurfactant detection was carried out using emulsification index, drop collapse and oil displacement assays. Functional composition of the crude biosurfactant was determined using gas chromatography-mass spectroscopy (GC-MS). In the presence of Pseudomonas fuscoginae significantly higher EI24 of 53.98% and 52.60% was observed in media that contained glucose or sodium acetate as carbon source, respectively. When P. fuscoginae or P. aeruginosa was used for inoculation, highest EI24 of 61.18% and 48.40% was observed in media that contained potassium nitrate as nitrogen source. In the medium that was inoculated with either B. subtilis or B. proteolyticus, highest EI24 of 53.65% and 49.63% was observed in the presence of tryptone and peptone, respectively. At the respective pH used for investigation, significantly highest EI24 was observed at pH 6, when inoculated with the respective isolates. In the case of incubation temperature, positive results were obtained throughout the incubation period at 25 and 30 oC. This was also irrespective of the isolate used for inoculation. All the extracted biosurfactants showed antimicrobial potentials against the test pathogens used for investigation. Characterization of the crude biosurfactant revealed the presence of compounds with antimicrobial properties. The study was able to provide useful information on optimum conditions for biosurfactant production by test bacterial species and the potential for possible application of the biosurfactants as antimicrobial agents.

Keywords: Biosurfactant, Surface tension, Emulsification, Bacterial species

The reliability of catechol 1,2-dioxygenase enzyme as detection factor of Pseudomonas savastanoi pv. savastanoi strains isolated from different olive growing areas in Jordan by PCR-RFLP

Bilal Ibrahim Wreikat*

Department of Plant Production and Protection, Faculty of Agricultural Technology, Al-Balqa Applied University, Al-Salt, Jordan

Abstract

The virulence gene Catechol 1,2-dioxygenase was detected in different isolates of Pseudomonas savastanoi pv. savastanoi (Smith, 1908), through amplification of 857 bp band by Polymerase Chain Reaction, it was confirmed in all isolates that were isolated from different olive cultivars growing in different areas in Jordan. Also, digestion of the amplified PCR product of this gene for all isolates of the pathogen, using Polymerase Chain Reaction Restriction-Fragments Length Polymorphism (PCR-RFLP), it was found that the catA gene is highly conserved for all isolates, after digestion with KpnI and BamHI Endonucleases. Further identification was performed for all isolates; by biochemical tests and pathogenicity on olive seedlings, and detection the virulence gene iaaL through PCR amplification of 454bp in all isolates. Interestingly, this study revealed that detection and identification of Pseudomonas savastanoi pv. savastanoi by catA gene is reliable and certified and will give further prospects in management between olive knot through crosstalk of olive plant and their knot bacterium.

Keywords: Olive Knot, Pseudomonas savastanoi, Catechol dioxygenase, Virulence factor

IAA-producing and phosphate solubilizer of rhizosphere actinobacteria consortium to promote plant growth in soybean (Glycine max L.)

Umi Fatmawati1*, Dewi Puspita Sari1, Slamet Santosa1, Sri Martina Wiraswati2

1Biology Education Study Program, Faculty of Teacher Training and Education, Universitas Sebelas Maret, Surakarta, Central Java, Indonesia

2Biology Study Program, Faculty of Biology, Universitas Jenderal Soedirman, Purwokerto, Central Java, Indonesia

Abstract

Plant Growth-Promoting Rhizobacteria (PGPR) are microbes that inhabit the rhizosphere and rhizoplanes environment and can enhance plant growth. The application of PGPR in agriculture can reduce the use of chemical fertilizers on soil. The purpose of this study was to determine the ability of the actinobacteria consortium to produce Indole-3-Acetic Acid (IAA), solubilize phosphate, and improve soybean growth in the greenhouse. The results of this study showed that the actinobacteria (Streptomyces sp. ASR58, Streptomyces sp. ASR67) and Rhizobium sp. which are used in this study are mutually compatible and tolerant to several types of fungicides and bactericides. Meanwhile, the consortium of Streptomyces sp. ASR58 and Streptomyces sp. ASR67 produces the highest concentration of IAA i.e. 25.11 ppm compared to each isolate and other bacterial consortia. Accordance to the quantitative phosphate solubilization assay, Streptomyces sp. ASR67 resulted in the highest dissolved inorganic phosphate i.e. 179.7 + 13.3 mg / L. Inoculation of Streptomyces sp. ASR58 and Streptomyces sp. ASR67 consortium into soybean seeds can significantly increase 54.6% in stem length, 29% root growth, and 20.4% in plant dry weight. This research indicated that Streptomyces sp. ASR58 and Streptomyces sp. ASR67 consortium resulted in the best growth toward soybean plants compared to other bacterial  consortia.

Keywords: Actinomycetes, IAA, Phosphate solubilization, Rhizosphere

Valorization of chicken feather into organic liquid fertilizer through two species of Bacillus bacteria fermentation 

Wardah1, Yasinta Lahum2, Feronika Fuakubun2, Tatang Sopandi2

1Study Program of Agroindustry, Vocational Faculty, Universitas 17 Agustus 1945, Surabaya, East Java, Indonesia

2Study Program of Biology, Faculty of Science and Technology, Universitas PGRI Adi Buana, Surabaya, East Java, Indonesia

 

Abstract

The valorization of industrial poultry waste by microbes into value-added products has received significant attention for reducing pollutants and producing a healthy environment. The current study aims to explore the use of liquid fertilizer from fermented chicken feathers by Bacillus cereus and Bacillus subtilis. The study was carried out experimentally ki using a completely randomized trial design with 10 different treatments of liquid fertilizer, each of which was repeated 12 times. The current study showed that both bacteria can secrete keratinase enzymes, degrade chicken feathers, and increase the nitrogen content of liquid fertilizer. However, the keratinase activity, degradation, and nitrogen content of chicken feather liquid fertilizer produced by B. subtilis fermentation were significantly higher (P˂0.05) than those of B. cereus. Likewise, the growth, anthocyanin content in the leaves, and yield of red spinach plants nurtured with chicken feather liquid fertilizer from B. subtilis fermentation were significantly (P˂0.05) higher than those given liquid fertilizer from B. cereus. The current study concluded that chicken feathers could be valorized into liquid fertilizer for plants, especially red spinach, by both B. cereus and B. subtilis bacteria. The current study recommends liquid fertilizer from chicken feathers fermented by B. subtilis at 0.05 g/plant to replace chemical fertilizers in plants, especially red spinach.

Keywords: Bacillus cereus, Bacillus subtilis, Chicken feather, Keratinase, Red spinach