02377nas a2200241   4500008004100000022001400041245011300055210006900168260007400237300001400311490000700325520145200332653004801784653001201832653002201844653002301866100001601889700001901905700002601924700002701950700002701977856013102004       2012                        eng d  a0301-485100aFunctional characterization and differential expression studies of squalene synthase from Withania somnifera0 aFunctional characterization and differential expression studies   aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcSEP  a8803-88120 v393 a<p>Squalene synthase (SQS: EC 2.5.1.21) is a potential branch point regulatory enzyme and represents the first committed step to diverge the carbon flux from the main isoprenoid pathway towards sterol biosynthesis. In the present study, cloning and characterization of Withania somnifera squalene synthase (WsSQS) cDNA was investigated subsequently followed by its heterologous expression and preliminary enzyme activity. Two different types of WsSQS cDNA clones (WsSQS1and WsSQS2) were identified that contained an open reading frames of 1,236 and 1,242 bp encoding polypeptides of 412 and 414 amino acids respectively. Both WsSQS isoforms share 99 % similarity and identity with each other. WsSQS deduced amino acids sequences, when compared with SQS of other plant species, showed maximum similarity and identity with Capsicum annuum followed by Solanum tuberosum and Nicotiana tabacum. To obtain soluble recombinant enzymes, 24 hydrophobic amino acids were deleted from the carboxy terminus and expressed as 6X His-Tag fusion protein in Escherichia coli. Approximately 43 kDa recombinant protein was purified using Ni-NTA affinity chromatography and checked on SDS-PAGE. Preliminary activity of the purified enzymes was determined and the products were analyzed by gas chromatograph-mass spectrometer (GC-MS). Quantitative real-time PCR (qRT-PCR) analysis showed that WsSQS expresses more in young leaves than mature leaves, stem and root.</p>10aGas chromatograph-Mass Spectrometer (GC-MS)10aqRT-PCR10aSqualene synthase10aWithania somnifera1 aGupta, Neha1 aSharma, Poonam1 aKumar, R., J. Santosh1 aVishwakarma, Rishi, K.1 aKhan, Bashir, Mohammad  uhttp://library.ncl.res.in/content/functional-characterization-and-differential-expression-studies-squalene-synthase-withania-001866nas a2200241   4500008004100000022001400041245011200055210006900167260000800236300001400244490000800258520105500266653002301321653001801344653001201362653002101374653002201395653002001417100001901437700001801456700002001474856013001494       2020                        eng d  a0888-754300aPlant-pathogen interactions: microRNA-mediated trans-kingdom gene regulation in fungi and their host plants0 aPlantpathogen interactions microRNAmediated transkingdom gene re  cSEP  a3021-30350 v1123 a<p>MicroRNAs (miRNAs) have been prevalently studied in plants, animals, and viruses. However, recent studies show evidences of miRNA-like RNAs (milRNAs) in fungi as well. It is known that after successful infection, pathogens hijack the host machinery and use it for their own growth and multiplication. Alternatively, resistant plants can overcome the pathogen attack by a variety of mechanisms. Based on this prior knowledge, we computationally predicted milRNAs from 13 fungi, and identified their targets in transcriptomes of the respective fungi as well as their host plants. The expressions of the milRNAs and targets were confirmed using qRT-PCR. We found that plant miRNAs targeted fungal virulence genes, while fungal milRNAs targeted plant resistance genes; corroborating miRNA-mediated trans-kingdom gene regulation and the roles of miRNAs in plantpathogen interactions. Transgenic plants with miRNAs targeting fungal virulence genes, or anti-sense of fungal milRNAs, would be expected to be highly resistant to the fungal pathogens.</p>
10aDisease resistance10aFungal miRNAs10aqRT-PCR10aResistance genes10aTarget prediction10aVirulence genes1 aMathur, Monika1 aNair, Aswathy1 aKadoo, Narendra  uhttp://library.ncl.res.in/content/plant-pathogen-interactions-microrna-mediated-trans-kingdom-gene-regulation-fungi-and-their