Identification and Characterization of microRNAs in the Developing Seed of Linseed Flax (Linum usitatissimum L.).

Seed development plays an important role during the life cycle of plants. Linseed flax is an oil crop and the seed is a key organ for fatty acids synthesis and storage. So it is important to understand the molecular mechanism of fatty acid biosynthesis during seed development. In this study, four small RNA libraries from early seeds at 5, 10, 20 and 30 days after flowering (DAF) were constructed and used for high-throughput sequencing to identify microRNAs (miRNAs). A total of 235 miRNAs including 114 known conserved miRNAs and 121 novel miRNAs were identified. The expression patterns of these miRNAs in the four libraries were investigated by bioinformatics and quantitative real-time polymerase chain reaction (qPCR) analysis. It was found that several miRNAs, including Lus-miRNA156a was significantly correlated with seed development process. In order to confirm the actual biological function of Lus-miRNA156a, over-expression vector was constructed and transformed to Arabidopsis. The phenotypes of homozygous transgenic lines showed decreasing of oil content and most of the fatty acid content in seeds as well as late flowering time. The results provided a clue that miRNA156a participating the fatty acid biosynthesis pathway and the detailed molecular mechanism of how it regulates the pathway needs to be further investigated.

Consensus genetic linkage map construction and QTL mapping for plant height-related traits in linseed flax (Linum usitatissimum L.).

BACKGROUND: Flax is an important field crop that can be used for either oilseed or fiber production. Plant height and technical length are important characters for flax. For linseed flax, plants usually have a short technical length and plant height than those for fiber flax. As an important agronomical character for fiber and linseed flax, plant height is usually a selection target for breeding. However, because of limited technologies and methods available, there has been little research focused on discovering the molecular mechanism controlling plant height. RESULTS: In this study, two related recombinant inbred line (RIL) populations developed from crosses of linseed and fiber parents were developed and phenotyped for plant height and technical length in four environments. A consensus linkage map based on two RIL populations was constructed using SNP markers generated by genotyping by sequencing (GBS) technology. A total of 4497 single nucleotide polymorphism (SNP) markers were included on 15 linkage groups with an average marker density of one marker every 2.71 cM. Quantitative trait locus (QTL) mapping analysis was performed for plant height and technical length using the two populations. A total of 19 QTLs were identified for plant height and technical length. For the MH population, eight plant height QTLs and seven technical length QTLs were identified, five of which were common QTLs for both traits. For the PH population, six plant height and three technical length QTLs were identified. By comparing the QTLs and candidate gene information in the two population, two common QTLs and three candidate genes were discovered. CONCLUSIONS: This study provides a foundation for map-based cloning of QTLs and marker-assisted selection for plant height-related traits in linseed and fiber flax.

Structure determination of archaea-specific ribosomal protein L46a reveals a novel protein fold.

Three archaea-specific ribosomal proteins recently identified show no sequence homology with other known proteins. Here we determined the structure of L46a, the most conserved one among the three proteins, from Sulfolobus solfataricus P2 using NMR spectroscopy. The structure presents a twisted ß-sheet formed by the N-terminal part and two helices at the C-terminus. The L46a structure has a positively charged surface which is conserved in the L46a protein family and is the potential rRNA-binding site. Searching homologous structures in Protein Data Bank revealed that the structure of L46a represents a novel protein fold. The backbone dynamics identified by NMR relaxation experiments reveal significant flexibility at the rRNA binding surface. The potential position of L46a on the ribosome was proposed by fitting the structure into a previous electron microscopy map of the ribosomal 50S subunit, which indicated that L46a contacts to domain I of 23S rRNA near a multifunctional ribosomal protein L7ae.

Identification and functional analysis of two alternatively spliced transcripts of ABSCISIC ACID INSENSITIVE3 (ABI3) in linseed flax (Linum usitatissimum L.).

Alternative splicing is a popular phenomenon in different types of plants. It can produce alternative spliced transcripts that encode proteins with altered functions. Previous studies have shown that one transcription factor, ABSCISIC ACID INSENSITIVE3 (ABI3), which encodes an important component in abscisic acid (ABA) signaling, is subjected to alternative splicing in both mono- and dicotyledons. In the current study, we identified two homologs of ABI3 in the genome of linseed flax. We screened two alternatively spliced flax LuABI3 transcripts, LuABI3-2 and LuABI3-3, and one normal flax LuABI3 transcript, LuABI3-1. Sequence analysis revealed that one of the alternatively spliced transcripts, LuABI3-3, retained a 6 bp intron. RNA accumulation analysis showed that all three transcripts were expressed during seed development, while subcellular localization and transgene experiments showed that LuABI3-3 had no biological function. The two normal transcripts, LuABI3-1 and LuABI3-2, are the important functional isoforms in flax and play significant roles in the ABA regulatory pathway during seed development, germination, and maturation.

Three new energetic complexes with N,N-bis(1H-tetrazole-5-yl)-amine as high energy density materials: syntheses, structures, characterization and effects on the thermal decomposition of RDX.

Three new energetic complexes, [Pb(bta)(H2O)]n (1), [PbCu(bta)2(H2O)5]·2H2O (2) and PbCu(bta)2 (3) (H2bta = N,N-bis(1H-tetrazole-5-yl)-amine), have been synthesized and characterised. In particular, 3 was readily synthesized by dehydration of 2 at 190 °C. Single crystal X-ray diffraction revealed that 1 has a 3D framework structure and 2 presents a 3D supermolecular architecture. Thermoanalyses demonstrated that the main frames of 1 and 2 have good thermostabilities up to 314 °C for 1 and 231 °C for 2. Non-isothermal kinetic and thermodynamic parameters of exothermic decomposition processes of 1 and 2 were obtained by Kissinger's and Ozawa's methods. Based on the constant-volume combustion energies measured by a precise rotating-bomb calorimeter, the standard molar enthalpies of formation of 1 and 2 were determined. The calculation of the detonation properties of 1 and 2 and the impact sensitivity tests of 1, 2 and 3 were carried out. In addition, 1, 2 and 3 were explored as combustion promoters to accelerate the thermal decompositions of RDX (1,3,5-trinitro-1,3,5-triazine) by differential scanning calorimetry. Experimental results showed that 1, 2 and 3 can be used as HEDMs in the field of combustion promoters and insensitive 2 can be regarded as a safer form for mass storage and transportation than sensitive 3.

Low stability of the reduced state of Mycobacterium tuberculosis NrdH redoxin.

NrdH redoxin is the only hydrogen donor for ribonucleotide reductase in Mycobacterium tuberculosis. Several crystal structures of NrdH redoxins in the oxidized state from different species have been reported, but no structure of the reduced state has yet been reported. Using NMR spectroscopy, we found surprisingly that the reduced NrdH redoxin from M. tuberculosis is largely unfolded at a pH lower than the pKa of its first active site cysteine, and the structural basis of the low stability was analyzed. In addition, a single mutant of the NrdH redoxin suitable to determine the structure in the reduced state was obtained.

Resonance assignments of a putative PilT N-terminus domain protein SSO1118 from hyperthermophilic archaeon Sulfolobus solfataricus P2.

PilT N-terminus (PIN) domains exist broadly in all three kingdoms of life, but the functions are not clear for most of them. Archaea species often encode multiple PIN domain-containing proteins, and the signaling and stress response roles have been proposed for these proteins. Some PIN domain proteins possess nuclease activities, which were proposed to be important in toxin-antitoxin stress response, nonsense-mediated mRNA decay, or RNA interference. SSO1118 from hyperthermophilic archaeon Sulfolobus solfataricus P2 is a putative PIN domain protein with low homology to other known PIN domain proteins. Here we report the NMR resonance assignments of SSO1118 for further structural determination and functional studies. The secondary structures predicted from the assigned chemical shifts consist with those of archaeal PIN domain proteins.