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Ziyu Dai, Shuang Deng, David E Culley, Kenneth S Bruno, Jon K Magnuson
Interest in using renewable sources of carbon, especially lignocellulosic biomass, for the production of hydrocarbon fuels and chemicals has fueled interest in exploring various organisms capable of producing hydrocarbon biofuels and chemicals or their precursors. The oleaginous (oil-producing) yeast Lipomyces starkeyi is the subject of active research regarding the production of triacylglycerides as hydrocarbon fuel precursors using a variety of carbohydrate and nutrient sources. The genome of L. starkeyi has been published, which opens the door to production strain improvements through the development and use of the tools of synthetic biology for this oleaginous species...
June 19, 2017: Applied Microbiology and Biotechnology
Haley M Ehrle, Jacob T Guidry, Rebecca Iacovetto, Anne K Salisbury, D J Sandidge, Grant Bowman
Despite being perceived as relatively simple organisms, many bacteria exhibit an impressive degree of sub-cellular organization. In Caulobacter crescentus, the evolutionarily conserved polar organizing protein PopZ facilitates cytoplasmic organization by recruiting chromosome centromeres and regulatory proteins to the cell poles. Here, we characterize the localization and function of PopZ in Agrobacterium tumefaciens, a genetically related species with distinct anatomy. In this species, we find that PopZ molecules are relocated from the old pole to the new pole in the minutes following cell division...
June 19, 2017: Journal of Bacteriology
Matthew Howell, Alena Aliashkevich, Anne K Salisbury, Felipe Cava, Grant R Bowman, Pamela J B Brown
Agrobacterium tumefaciens is a rod-shaped bacterium that grows by polar insertion of new peptidoglycan during cell elongation. As the cell cycle progresses, peptidoglycan synthesis at the pole ceases prior to insertion of new peptidoglycan at mid-cell to enable cell division. The A. tumefaciens homolog of the Caulobacter crescentus polar organelle development protein PopZ has been identified as a growth pole marker and a candidate polar growth promoting factor. Here, we characterize the function of PopZ in cell growth and division of A...
June 19, 2017: Journal of Bacteriology
Jun Yan, Yan Li, Hui Yan, Wen Feng Chen, Xiaoxia Zhang, En Tao Wang, Xiao Zeng Han, Zhi Hong Xie
Two Gram-staining-negative, aerobic bacteria (YIC 5082T and YIC4104) isolated from root nodules of Sesbania cannabina grown in a high-salt and alkaline environment were identified as a group in the genus Agrobacterium because they shared 100 and 99.7 % sequence similarities of 16S rRNA and recA+atpD genes, respectively. These two strains showed 99.2/100 % and 93.9/95.4 % 16S rRNA and recA+atpD gene sequence similarities to Agrobacterium radiobacter LMG140T and Agrobacterium. pusense NRCPB10T, respectively...
June 20, 2017: International Journal of Systematic and Evolutionary Microbiology
Wenjun Yu, Rongshui Wang, Haiyan Huang, Huijun Xie, Shuning Wang
Agrobacterium tumefaciens S33 can grow with nicotine as the sole source of carbon, nitrogen, and energy via a novel hybrid of the pyridine pathway and the pyrollidine pathway. Characterization of the enzymes involved in the hybrid pathway is important for understanding its biochemical mechanism. Here we report that the molybdenum-containing nicotine dehydrogenase (NdhAB), which catalyzes the initial step of nicotine degradation, is located in the periplasm of strain S33, while the 6-hydroxynicotine oxidase and 6-hydroxypseudooxynicoine oxidase are in the cytoplasm...
June 16, 2017: Applied and Environmental Microbiology
Fahimeh Nazari, Naser Safaie, Bahram Mohammad Soltani, Masoud Shams-Bakhsh, Mohsen Sharifi
Agrobacterium tumefaciens is a very destructive plant pathogen. Selection of effective biological agents against this pathogen depends on more insight into molecular plant defence responses during the biocontrol agent-pathogen interaction. Auxin as a phytohormone is a key contributor in pathogenesis and plant defence and accumulation of auxin transport carriers are accompanied by increasing in flavonoid and miRNAs concentrations during plant interactions with bacteria. The aim of this research was molecular analysis of Bacillus subtilis (ATCC21332) biocontrol effect against A...
June 10, 2017: Plant Physiology and Biochemistry: PPB
Sheng Xu, Bin Sun, Rong Wang, Jia He, Bing Xia, Yong Xue, Ren Wang
The phytoremediation by using of green plants in the removal of environmental pollutant is an environment friendly, green technology that is cost effective and energetically inexpensive. By using Agrobacterium-mediated gene transfer, we generated transgenic Arabidopsis plants ectopically expressing mercuric transport protein gene (merT) from Pseudomonas alcaligenes. Compared with wild-type (WT) plants, overexpressing PamerT in Arabidopsis enhanced the tolerance to HgCl2. Further results showed that enhanced total activities or corresponding transcripts of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (POD) were observed in transgenic Arabidopsis under HgCl2 stress...
June 14, 2017: Biochemical and Biophysical Research Communications
C Cosseau, S Romano-Bertrand, H Duplan, O Lucas, I Ingrassia, C Pigasse, C Roques, E Jumas-Bilak
The human skin microbiota is quantitatively dominated by Gram-positive bacteria, detected by both culture and metagenomics. However, metagenomics revealed a huge variety of Gram-negative taxa generally considered from environmental origin. For species affiliation of bacteria in skin microbiota, clones of 16S rRNA gene and colonies growing on diverse culture media were analyzed. Species-level identification was achieved for 81% of both clones and colonies. Fifty species distributed in 26 genera were identified by culture, mostly belonging to Actinobacteria and Firmicutes, while 45 species-level operational taxonomic units distributed in 30 genera were detected by sequencing, with a high diversity of Proteobacteria...
December 2016: One Health
Meizhen Wang, Yuxing Deng, Fenjuan Shao, Miaomiao Liu, Yongqi Pang, Caili Li, Shanfa Lu
Small RNA-mediated gene silencing is a vital regulatory mechanism in eukaryotes that requires ARGONAUTE (AGO) proteins. Salvia miltiorrhiza is a well-known traditional Chinese medicinal plant. Therefore, it is important to characterize S. miltiorrhiza AGO family genes as they may be involved in multiple metabolic pathways. This chapter introduces the detailed protocol for SmAGO gene prediction and molecular cloning. In addition, an Agrobacterium-mediated genetic transformation method for S. miltiorrhiza is presented...
2017: Methods in Molecular Biology
Hong-Xian Hsing, Yi-Jyun Lin, Chii-Gong Tong, Min-Jeng Li, Yun-Jin Chen, Swee-Suak Ko
BACKGROUND: Phalaenopsis orchid (Phal. orchid) is visually attractive and it is important economic floriculture species. Phal. orchids have many unique biological features. However, investigation of these features and validation on their biological functions are limited due to the lack of an efficient transformation method. RESULTS: We developed a heritable and efficient Agrobacterium- mediated transformation using protocorms derived from tetraploid or diploid Phal...
December 2016: Botanical Studies (Taipei, Taiwan)
Jessica Purswani, Rocío C Romero-Zaliz, Antonio M Martín-Platero, Isabel M Guisado, Jesús González-López, Clementina Pozo
Ecosystem functionality depends on interactions among populations, of the same or different taxa, and these are not just the sum of pairwise interactions. Thus, know-how of the social interactions occurring in mixed-populations are of high interest, however they are commonly unknown due to the limitations posed in tagging each population. The limitations include costs/time in tediously fluorescent tagging, and the number of different fluorescent tags. Tag-free strategies exist, such as high-throughput sequencing, but ultimately both strategies require the use of expensive machinery...
2017: Frontiers in Microbiology
Chong Zhang, Songxiao Cao, Yazhong Jin, Lijun Ju, Qiang Chen, Qiaojuan Xing, Hongyan Qi
To better understand the function role of the melon CmLOX18 gene in the biosynthesis of C6 volatiles during fruit ripening, we biochemically characterized CmLOX18 and identified its subcellular localization in transgenic tomato plants. Heterologous expression in yeast cells showed that the molecular weight of the CmLOX18 protein was identical to that predicted, and that this enzyme possesseed lipoxygenase activity. Linoleic acid was demonstrated to be the preferred substrate for the purified recombinant CmLOX18 protein, which exhibited optimal catalytic activity at pH 4...
June 6, 2017: Scientific Reports
Le Kang, Chang Yoon Ji, Sun Ha Kim, Qingbo Ke, Sung-Chul Park, Ho Soo Kim, Hyeong-Un Lee, Joon Seol Lee, Woo Sung Park, Mi-Jeong Ahn, Haeng-Soon Lee, Xiping Deng, Sang-Soo Kwak
β-carotene, a carotenoid that plays a key photo-protective role in plants is converted into zeaxanthin by β-carotene hydroxylase (CHY-β). Previous work showed that down-regulation of IbCHY-β by RNA interference (RNAi) results in higher levels of β-carotene and total carotenoids, as well as salt stress tolerance, in cultured transgenic sweetpotato cells. In this study, we introduced the RNAi-IbCHY-β construct into a white-fleshed sweetpotato cultivar (cv. Yulmi) by Agrobacterium-mediated transformation...
May 29, 2017: Plant Physiology and Biochemistry: PPB
Philippa J Barrell, Julie M Latimer, Samantha J Baldwin, Michelle L Thompson, Jeanne M E Jacobs, Anthony J Conner
BACKGROUND: Somatic cell selection in plants allows the recovery of spontaneous mutants from cell cultures. When coupled with the regeneration of plants it allows an effective approach for the recovery of novel traits in plants. This study undertook somatic cell selection in the potato (Solanum tuberosum L.) cultivar 'Iwa' using the sulfonylurea herbicide, chlorsulfuron, as a positive selection agent. RESULTS: Following 5 days' exposure of potato cell suspension cultures to 20 μg/l chlorsulfuron, rescue selection recovered rare potato cell colonies at a frequency of approximately one event in 2...
June 6, 2017: BMC Biotechnology
Tagginahalli N Shivakumara, Rohini Sreevathsa, Prasanta K Dash, M S Sheshshayee, Pradeep K Papolu, Uma Rao, Narendra Tuteja, M UdayaKumar
Imparting tolerance to abiotic stresses is of global importance as they inflict significant yield losses in field as well as in vegetable crops. Transcriptional activators, including helicases are identified to play a pivotal role in stress mitigation. Helicases, also known as molecular motors, are involved in myriad cellular processes that impart intrinsic tolerance to abiotic stresses in plants. Our study demonstrates the potential of a Pea DNA Helicase 45 (PDH45), in combating multiple abiotic stresses in chili...
June 5, 2017: Scientific Reports
Liping Ding, Yajuan Chen, Xiaoli Wei, Mi Ni, Jiewei Zhang, Hongzhi Wang, Zhen Zhu, Jianhua Wei
Transgenic poplar lines 'Shanxin' (Populus davidiana×Populus bolleana) were generated via Agrobacterium-mediated transformation. The transgenic lines carried the expression cassettes of Cry1Ac + SCK, Cry1Ah3, and Cry9Aa3, respectively. The expression levels of the exogenous insect resistance genes in the transgenic lines were determined by Q-PCR and Western blot. Leaves of the transgenic lines were used for insect feeding bioassays on first instar larvae of the gypsy moth (Lymantria dispar) and fall webworm (Hyphantria cunea)...
2017: PloS One
Shujuan Li, Cheng Zhen, Wenjing Xu, Chong Wang, Yuxiang Cheng
Genotype Nisqually-1 is the first model woody plant with an available well-annotated genome. Nevertheless, a simple and rapid transformation of Nisqually-1 remains to be established. Here, we developed a novel shoot regeneration method for Nisqually-1 using leaf petiole and stem segment explants. Numerous shoots formed in the incision of explants within two weeks. The optimized shoot regeneration medium (SRM) contained 0.03 mg l(-1) 6-benzylaminopurine, 0.02 mg l(-1) indole-3-butyric acid and 0.0008 mg l(-1) thidiazuron...
June 1, 2017: Scientific Reports
Skylar L Fuller, Elizabeth Savory, Alexandra J Weisberg, Jessica Z Buser, Michael I Gordon, Melodie Putnam, Jeff H Chang
Agrobacterium is a genus of soil-borne Gram-negative bacteria. Members carrying oncogenic plasmids can cause crown gall disease, which has significant economic costs, especially for the orchard and nursery industries. Early and rapid detection of pathogenic Agrobacterium is key to the management of crown gall disease. To this end, we designed oligonucleotide primers and probes to target virD2 for use in a molecular diagnostic tool that relies on isothermal amplification and lateral flow-based detection. The oligonucleotide tools were tested in the assay and evaluated for sensitivity, specificity, and breadth in detecting alleles of virD2...
June 1, 2017: Phytopathology
Fengqing Wang, Jingyu Zhi, Zhongyi Zhang, Lina Wang, Yanfei Suo, Caixia Xie, Mingjie Li, Bao Zhang, Jiafang Du, Li Gu, Hongzheng Sun
Rehmannia glutinosa is a common bulk medicinal material that has been widely used in China due to its active ingredients. Acteoside, one of the ingredients, has antioxidant, antinephritic, anti-inflammatory, hepatoprotective, immunomodulatory, and neuroprotective effects, is usually selected as a quality-control component for R. glutinosa herb in the Chinese Pharmacopeia. The acteoside biosynthesis pathway in R. glutinosa has not yet been clearly established. Herein, we describe the establishment of a genetic transformation system for R...
2017: Frontiers in Plant Science
Michelle F Valentine, Joann R De Tar, Muruganantham Mookkan, Jeffre D Firman, Zhanyuan J Zhang
Soybean [Glycine max (L.) Merr.] is the number one oil and protein crop in the United States, but the seed contains several anti-nutritional factors that are toxic to both humans and livestock. RNA interference technology has become an increasingly popular technique in gene silencing because it allows for both temporal and spatial targeting of specific genes. The objective of this research is to use RNA-mediated gene silencing to down-regulate the soybean gene raffinose synthase 2 (RS2), to reduce total raffinose content in mature seed...
2017: Frontiers in Plant Science
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