02469nas a2200253   4500008004100000022001400041245011400055210006900169260007400238300001200312490000700324520154700331653000901878653002901887653003101916653000801947100002101955700002501976700002402001700002602025700002102051700002102072856012202093       2012                        eng d  a1380-374300aDevelopment of genomic simple sequence repeat markers for linseed using next-generation sequencing technology0 aDevelopment of genomic simple sequence repeat markers for linsee  aVAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDSbSPRINGERcJUN  a597-6060 v303 a<p>Linseed (Linum usitatissimum L.) is regarded as a cash crop of tomorrow because of the presence of nutraceutically important alpha-linolenic acid (ALA) and lignan. However, only limited breeding progress has been made in this crop, mainly due to the lack of sufficient genetic and genomic resources. Among these, simple sequence repeats (SSR) are useful DNA markers for diversity analysis, genetic mapping and tagging traits because of their co-dominant and highly polymorphic nature. In order to develop SSR markers for linseed, we used three microsatellite isolation methods, viz., PCR Isolation of Microsatellite Arrays (PIMA), 5'-anchored PCR method, and Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO). The amplified products from these methods were pooled and sequenced using the 454 GS-FLX platform. A total of 36,332 reads were obtained, which assembled into 2,183 contigs and 2,509 singlets. The contigs and the singlets contained 1,842 microsatellite motifs, with dinucleotide motifs as the most abundant repeat type (54%) followed by trinucleotide motifs (44%). Based on this, 290 SSR markers were designed, 52 of which were evaluated using a panel of 27 diverse linseed genotypes. Among the three enrichment methods, the 5'-anchored PCR method was most efficient for isolation of microsatellites, while FIASCO was most efficient for developing SSR markers. We show the utility of next-generation sequencing technology for efficiently discovering a large number of microsatellite markers in non-model plants.</p>10aFlax10aMicrosatellite isolation10aNext-generation sequencing10aSSR1 aKale, Sandip, M.1 aPardeshi, Varsha, C.1 aKadoo, Narendra, Y.1 aGhorpade, Prakash, B.1 aJana, Murari, M.1 aGupta, Vidya, S.  uhttps://library.ncl.res.in/content/development-genomic-simple-sequence-repeat-markers-linseed-using-next-generation-002740nas a2200277   4500008004100000022001400041245017800055210006900233260010600302300001000408490000700418520166900425653000802094653002702102653001602129653000902145653002002154653001202174100002502186700002402211700002402235700002502259700002602284700002102310856013102331       2014                        eng d  a0031-942200aDifferential transcriptional activity of SAD, FAD2 and FAD3 desaturase genes in developing seeds of linseed contributes to varietal variation in alpha-linolenic acid content0 aDifferential transcriptional activity of SAD FAD2 and FAD3 desat  aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcFEB  a41-530 v983 a<p>Linseed or flax (Linum usitatissimum L.) varieties differ markedly in their seed a-linolenic acid (ALA) levels. Fatty acid desaturases play a key role in accumulating ALA in seed. We performed fatty acid (FA) profiling of various seed developmental stages of ten Indian linseed varieties including one mutant variety. Depending on their ALA contents, these varieties were grouped under high ALA and low ALA groups. Transcript profiling of six microsomal desaturase genes (SAD1, SAD2, FAD2, FAD2-2, FAD3A and FAD3B), which act sequentially in the fatty acid desaturation pathway, was performed using real-time PCR. We observed gene specific as well as temporal expression pattern for all the desaturases and their differential expression profiles corresponded well with the variation in FA accumulation in the two groups. Our study points to efficient conversion of intermediate FAs [stearic (SA), oleic (OA) and linoleic acids (LA)] to the final product, ALA, due to efficient action of all the desaturases in high ALA group. While in the low ALA group, even though the initial conversion up to OA was efficient, later conversions up to ALA seemed to be inefficient, leading to higher accumulation of OA and LA instead of ALA. We sequenced the six desaturase genes from the ten varieties and observed that variation in the amino acid (AA) sequences of desaturases was not responsible for differential ALA accumulation, except in the mutant variety TL23 with very low (&lt;2%) ALA content. In TL23, a point mutation in the FAD3A gene resulted into a premature stop codon generating a truncated protein with 291 AA. (C) 2013 Elsevier Ltd. All rights reserved.</p>10aALA10aFatty acid desaturases10aFatty acids10aFlax10aGene expression10aLinseed1 aRajwade, Ashwini, V.1 aKadoo, Narendra, Y.1 aBorikar, Sanjay, P.1 aHarsulkar, Abhay, M.1 aGhorpade, Prakash, B.1 aGupta, Vidya, S.  uhttps://library.ncl.res.in/content/differential-transcriptional-activity-sad-fad2-and-fad3-desaturase-genes-developing-seeds-0