First Report of Leek yellow stripe virus in Allium sativum in Western India.
2区 · 农林科学
作者: S J Gawande ; V S Gurav ; A A Ingle ; J Gopal
Garlic (Allium sativum L.) is an important bulbous spice crop in India as well as other parts of world. Garlic is well known for its medicinal properties. Degeneration due to viral infections is one of the important constraints in exploiting its yield potential. Leek yellow stripe virus (LYSV), genus Potyvirus, family Potyviridae, is a prominent virus known to infect garlic worldwide (4). During July 2013, potyvirus-like symptoms such as mosaic, streaking, stunting, mottling of leaves were observed on garlic cv. G-41 and landrace Ranibennur local, collected from Karnataka, India, and maintained at the Directorate of Onion and Garlic Research, Rajgurunagar, Pune, India. The incidence of symptomatic plants was estimated at 70% for Ranibennur local and 68% for cv. G-41. The symptomatic leaves were sampled diagonally from the field. Twenty symptomatic plants per cultivar with each sample was composited from young, middle, and lower (basal) leaves of the plant. These samples were tested by double-antibody sandwich (DAS)-ELISA for LYSV using commercially available kit (Agdia Inc., Elkhart, IN). ELISA-positive plants were further subjected to molecular studies. Total RNA from the infected leaf samples were extracted by RNeasy Plant Mini kit (Qiagen GmbH, Hilden, Germany) and assayed by reverse transcription (RT)-PCR using primer pair LYSV-F 2 (5'-GCACCATACAGTGAATTGAG-3') (1), LYSV-R (5'-GCCTCGCGCGCTCTAA-3') (3) to amplify 874 bases of partial Nib and partial coat protein gene. The amplified product of 874 bp derived from A. sativum isolate was purified (QIAquick PCR Purification Kit, Qiagen) and cloned using vector pDrive (Qiagen). The recombinant clones were sequenced and submitted in NCBI database (GenBank Accession No. KF850539). The sequence analysis performed on CLC Main Workbench Version 6.8.4 gave confirmation of LYSV. Further, phylogenetic analysis of the 874-nt sequence revealed the highest nucleotide identity (80 to 82%) with LYSV isolates (DQ925453, JN127339, AB005611, and JX429965). To the best of our knowledge, this is the first report of natural infection of garlic by LYSV in western India. LYSV is known to cause direct losses in garlic and other related Allium spp. Up to 54% reduction in bulb weight was observed due to single infection of this virus (2). Hence, our first report about this virus has significant impact on garlic production scenario, if this virus found to be widespread in the country. For this, additional surveys and genotype screenings are needed to obtain a better understanding of the potential impact of LYSV on garlic production in India. References: (1) H. Fidan and S. Baloglu. Plant Dis. 93:672, 2009. (2) H. Lot et al. Plant Dis. 82:1381, 1998. (3) P. Lunello et al. J. Virol. Methods. 118:15, 2004. (4) H. R. Pappu et al. Plant Dis. 89:205, 2005.
2010-08-01·Plant disease2区 · 农林科学
First Report of Iris yellow spot virus on Garlic in India.
2区 · 农林科学
作者: S J Gawande ; A Khar ; K E Lawande
Garlic (Allium sativum) is a spice crop of prime importance in India as well as other parts of the world. Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) is an important pathogen of onion bulb and seed crops in many parts of the world (3). The virus is also known to infect garlic and other Allium spp. (2-4). IYSV infection of garlic was reported from Reunion Island (4) and the United States (1). In February 2010, straw-colored, spindle-shaped spots with poorly defined ends were observed on the leaves of a garlic crop at the research farm of the Directorate of Onion and Garlic Research in the Pune District of Maharashtra State, India, 105 days after planting. The spots coalesced to form larger patches on the leaves, suggesting possible IYSV infection. Symptoms were visible on older leaves and more prevalent on cv. G-41, G-282, AC50, AC200, AC283, and Godavari than on other cultivars. The incidence of symptomatic plants was estimated at 5% for G-41 and AC-200, 8% for G-282 and AC283, and 10% for AC50. Leaves were sampled from 40 symptomatic plants per cultivar with each sample composited from young, middle, and older (basal) leaves of the plant. Samples were assayed by double-antibody sandwich-ELISA (Loewe Biochemica GmbH, Sauerlach, Germany) and each tested positive for the virus. Total RNA was extracted from the leaves of ELISA-positive plants using the RNAeasy Plant Mini kit (Qiagen GmbH, Hilden, Germany) and tested by reverse transcription-PCR assay using primers IYSV-F (5'-TCAGAAATCGAGAAACTT-3') and IYSV-R (5'-TAATTATATCTATCTTTCTTGG-3') (2) designed to amplify 797 bp of the nucleocapsid (N) gene of IYSV. Amplicons of expected size were obtained and cloned into a pDrive vector (Qiagen GmbH). The recombinant clone was sequenced (GenBank Accession No. HM173691). Sequence comparisons showed 98 to 100% nt identity with other IYSV N gene sequences in GenBank (Nos. EU310294 and EU310286). A phylogenetic analysis of the deduced amino acid sequences of the N gene showed that the garlic isolate of IYSV grouped most closely with onion IYSV isolates from India (GenBank Nos. EU310294, EU310286, EU310300, and EU310296). To our knowledge, this is the first report of natural infection of garlic by IYSV in India. Additional surveys and evaluations are needed to obtain a better understanding of the potential impact of IYSV on garlic production in India. References: (1) S. Bag et al. Plant Dis. 93:839, 2009. (2) A. Bulajic et al. Plant Dis. 93:976, 2009. (3) D. Gent et al. Plant Dis. 90:1468, 2006. (4) I. Robène-Soustrade et al. Plant Pathol. 55:288, 2006.
2003-12-01·Journal of Steroid Biochemistry and Molecular Biology2区 · 生物学
Production of malodorous steroids from androsta-5,16-dienes and androsta-4,16-dienes by Corynebacteria and other human axillary bacteria
2区 · 生物学
作者: Decreau, Richard A. ; Marson, Charles M. ; Smith, Kelvin E. ; Behan, John M.
The biotransformations of a number of steroids, chiefly 5,6,16,17-tetradehydro-androstanes, are reported. The strains investigated were Corynebacteria sp. G38, G40, G41, B, Brevis sp. CW5 and Micrococcus sp. M-DH2. Corynebacterium sp. G41 proved remarkably efficient in effecting oxidative isomerisation of 5-ene-3-sterols into the corresponding 4-en-3-ones. The main biochemical reactions involved were oxidation at C-3; no reduction processes were observed. Conversions of 3beta-sterols into the C-3 oxo-steroids were high, but were correspondingly low for the 3alpha-sterol epimers. Androsta-4,16-dien-3-one and 5beta-androsta-16-en-3-one are crucial to the formation of malodour. The rate of formation of these compounds was measured over 72 h incubation periods using three substrates: androsta-5,16-dien-3beta-ol, androsta-4,16-dien-3beta-ol and androsta-5,16-dien-3-one. Induction studies of the transformation of the androsta-5,16-dien-3beta-ol into the very odorous compound androsta-4,16-dien-3-one showed that cells incubated with a mixture of antibiotics displayed the same extent of biotransformation as normal cells if the concentration of antibiotic was low (1, 3, 5 and 7 microg/ml), although at concentrations higher than 10 microg/ml, biotransformation yields were reduced. Pre-incubation with a 3beta-fluoro-steroid inhibited the formation of the odorous androsta-4,16-dien-3-one.
Produces 98% pure protein
THOUSAND OAKS, Calif., Feb. 24 /PRNewswire/ -- Bone Biologics, Inc. announced today that it's contract research and development organization, Aragen Bioscience, has completed development of a recombinant cell line and initial production process for UCB-1 (NELL-1) recombinant protein.
Bone Biologics has been developing the protein as a platform technology since 2004, leveraging the previous ten years of research in the lab performed by the founding scientists at UCLA. The mechanism of action is not only identified as to how it works, but why, through this exhaustive research effort. This platform technology is combined with DBX(R) demineralized bone matrix to promote bone growth in spinal fusion.
Aragen has produced UCB-1 (NELL-1) at a purity level Bone Biologics believes is sufficient for IDE submission. Results in animal trials using that protein have exhibited rapid bone growth, high quality of bone, and no ectopic bone growth. Efforts will now focus on scaling that process up to produce clinical grade product.
The platform technology will be subject to the FDA review and approval process, including clinical trials. It is not currently approved for use in humans.
Rick Srigley, CEO of Aragen Bioscience said, "This project has been exciting for us. UCB-1 (NELL-1) is a complex and challenging protein, but we believe that the cell line and process we have developed for Bone Biologics can be scaled up to provide protein at purity and productivity levels sufficient for Phase I clinical studies." William Jay Treat, Ph.D., President of Southwest BioProcessing Associates LLC, who has extensive experience in cell line development, scale-up optimization and commercial manufacturing provided key guidance during this process to Bone Biologics, said, "The professionalism, expertise, and awareness of keeping to a timeline by Aragen has developed a manufacturing process that is suitable to move to the next phase of UCB-1 (NELL-1) manufacturing.
About Aragen Biosciences
Aragen Biosciences, Inc. is a contract research and development organization in Morgan Hill, California. Aragen specializes in recombinant cell line and hybridoma development, cell culture and purification process development and production, cell-based assays and custom immunological services. ( )
About Bone Biologics, Inc.
Bone Biologics is a convergence biotechnology company focused in the orthobiologics space. The company was founded in 2004 by the research scientists who had identified and validated the product in the lab over the previous ten years. This platform technology is the unique UCB-1 (NELL-1) protein formulated with tissue-specific scaffolds to only deliver the intended outcomes. The lead product effort is to promote bone growth in spinal fusion.
CONTACT: Bruce A. Hazuka of Bone Biologics, +1-818-324-2742