Plant-parasitic nematodes are the most devastating group of plant pathogens worldwide and are extremely challenging to control. In the present study, we have performed a genome wide analysis to identify common genes among four nematode species consisting of root-knot nematodes (Meloidogyne incognita and Meloidogyne hapla), cyst nematode (Heterodera glycines), and free living nematode (Caenorhabditis elegans) respectively. Using their whole genome sequences, we predicted 15,274 genes from M. incognita, 38,149 genes from M. hapla, 8,061 genes from H. glycines and 23,894 genes from C. elegans, where, among the predicted genes, 1,358, 1,350, 1,401, 1,365 respectively from each nematode, code for common groups of proteins. Further, 2,067, 2,086, 1,566, 2,903 genes were recollected using Clusters of Orthologous Groups (COG) database. Under our search criteria, a total of 800 common genes were identified in all the four studied nematode genomes. The most annotated conserved genes were obtained from four different species using Basic Local Alignment Searching Tool (BLAST). Uni- Prot Taxon identifier database was used to elucidate their taxonomic classification such as 698 genes under kingdom Metazoa, 660 genes confined to Nematoda, 290 genes in Chordata and 660 genes falling under class Chromadorea. The biochemical characterization of proteins expressed by these genes was examined using Pedant-Pro sequence analysis. The protein length, molecular weight, isoelectric point (pI), and transmembrane domain of the coded proteins were at a range of 300 to 999 amino acids (40.9%), molecular weight of over 100 kDa (96%), pI from 4.5 to 5.5 (27.6%) and 0 (56.6%), respectively. To classify protein function, the obtained BLAST hits were assigned to Gene Ontology classification scheme. The fractions of protein function were distributed as cellular component, biological processes and molecular function of the cell (22.2%), multicellular organism process (15.8%) and binding (48.3%), respectively. The current study provides an excellent resource for nematode functional genomics studies, which can be utilized further for studies on role of genes involved in nematode biological processes.
The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges. fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylationsites(Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1(Asn-454), fvLac-2(Asn- 437andAsn-455), fvLac-3(Asn-111andAsn-237), and fvLac4 (Asn-402andAsn-457). In addition, the first 19–20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction(RT-PCR) analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.
The 35.6-mb genome contained 12,218 predicted protein-encoding genes and 287 tRNA genes assembled into 11 scaffolds corresponding with the 11 chromosomes of strain KACC 42780. Based on presence of copper binding region and signal peptide, 8 laccase genes (Fv Lac-1, Fv Lac-2, Fv Lac-3, Fv Lac-4, Fv Lac-5, Fv Lac-6, Fv Lac-7, Fv Lac-8) were selected in F.velutipes genome. The laccase genes of F.velutipes ranged from 1,300 to 1,700 base pair in their size. In addition, the molecular mass and PI values of laccase genes ranged from 50 to 62 kDa and from 3 to 11, respectively.
In laccase activity assays, the highest laccase activity (3unit/ml)was shown in medium with 0.5mM of CuSO4 on 9th day. However, laccase activity was severely decreased in medium with 1mM and 2mM of CuSO4 was drastically decreased from day 3 through day 9. Interestingly, laccase activity in the medium without CuSO4 showed higher level than in the medium with 0.25, 1 and 2mM on both 6th and 9th day.
RT_PCR showed the highest transcription level with 0.25mM and 0.5mM of CuSO4 supplementation on day 3 and on day 9, respectively. In addition, the transcription level of Lac-1 increased depending on concentration of CuSO4 supplementation.
Sessile organism, plants constitutively challenged with pathogens have been developed various strategies for protection, such as preformed and inducible defense mechanisms. Receptor-like Proteins(RLPs) play critical roles in defense response as well as in plant development and growth. The domain structure of RLPs consists of extracellular leucine–rich repeats, a transmembrane domain, and a short cytoplasmic tail. Here, we identified putative 170 RLP genes from pepper genome using in-house bioinformatics pipeline. The distribution of RLPs on pepper pseudomolecule showed uneven spread and a number of RLPs were physically clustered by tandem array in the specific chromosome. Motifs analysis of pepper RLPs showed conserved LRR sequences (LxxLxxLDLxxNxxxGxIP). To understand further functional and evolutionary characteristics, evolutional relationship and gene profiling analysis are on progress.
Plants have evolved elaborate innate immune systems against invading pathogens, such as bacteria, fungi, oomycetes, viruses and insects. Among them, intracellular immune receptors known as nucleotide-binding site and leucine-rich repeat (NB-LRR) play critical roles in effector-triggered immunity (ETI) regarding to plant defense. Here, we identified potential NB-LRR coding sequences from pepper genome using bioinformatics analysis and performed comparative analysis with Solanaceae plants. As a result, we identified 267, 443, and 755 NBS-encoding genes in the genome of tomato, potato, and pepper, respectively. These may indicate that the Solanaceae NB-LRRs were evolved through species-specific unequal-duplication event. Further phylogenetic and clustering analyses revealed that Solanaceae NB-LRRs were classified into the 14 subgroups with 1 TNL and 13 CNL types. We found that the genes in CNL-G1 and CNL-G2 subgroup were highly expanded compared to other subgroup showing a large portion of NB-LRR in pepper genome. Among 755 NB-LRRs in pepper genome, 623 were physically mapped on all 12 pepper chromosome pseudomolecules. Furthermore, a number of NB-LRRs in the same group were physically clustered by tandem array in the specific chromosome. Genome-wide identification of pepper NB-LRR family and their evolutionary analysis could provide an important resource for identification and characterization of genes for breeding of disease resistance crops.
sequence and more than fifty thousand proteins have been obtained to date. Transcription factors (TFs) are important regulators involved in plant development and physiological processes and the AP2/ERF protein family contains TFs that also plays a crucial role as well and response to biotic and abiotic stress conditions in plants. However, no detailed expression profile of AP2/ERF-like genes is available for B. oleracea. In the present study, 226 AP2/ERF TFs were identified from B. oleracea based on the available genome sequence. Based on sequence similarity, the AP2/ERF superfamily was classified into five groups (DREB, ERF, AP2, RAV and Soloist) and 15 subgroups. The identification, classification, phylogenetic construction, conserved motifs, chromosome distribution, functional annotation, expression patterns and interaction network were then predicted and analyzed. AP2/ERF TFs expression levels exhibited differences in response to varying abiotic stresses based on expressed sequence tags (ESTs). BoCBF1a, 1b, 2, 3 and 4, which were highly conserved in Arabidopsis and B. rapa CBF/DREB genes families were well characterized. Expression analysis enabled elucidation of the molecular and genetic level expression patterns of cold tolerance (CT) and susceptible lines (CS) of cabbage and indicated that all BoCBF genes responded to abiotic stresses. Comprehensive analysis of the physiological functions and biological roles of AP2/ERF superfamily genes and BoCBF family genes in B. oleracea is required to elucidate AP2/ERF, which will provide rich resources and opportunities to understand abiotic stress tolerance in crops.
Sequence diversity was accumulated through evolution and breeding process. A set of 595 PCR-based novel insertion/deletion (InDel) markers was designed in order to widen the genetic basis for national rice breeding programs. The markers were generated by analyzing of 40 Korean cultivars and published genome sequences of rice(Oryza sativa L. spp japonica). We selected 112 markers spread across all rice chromosomes among the 595 InDel markers, and they showed polymorphic between rice cultivars, which are 284 Korean japonica and Tongil varieties. Due to their simplicity in design and robustness in genotyping, these InDel markers have been routinely used in quantitative trait loci (QTL) mapping studies and marker assisted selection programs for rice. Moreover, the PCR amplification type of InDel markers was converged to digital code, 0 or 1 and then finally represented as one- and two dimensional bar-code system, which could easily differentiate genetically highly homologous japonica rice cultivars. The developed InDel markers uniquely discriminated among each of the Korean cultivars. Therefore, the systems we developed may be valuable tools in discrimination from cultivars
Spatial- and temporal-specific expression patterns are primarily regulated at the transcriptional level by the promoter. Therefore, it is important to determine the binding motifs of transcription factors to understand the networks associated with embryogenesis. Here, we used a protein-binding microarray (PBM) to determine the binding motif of OsSMF1, which is a basic leucine zipper transcription factor that is involved in the regulation of rice seed maturation. OsSMF1 (previously called RISBZ1) is known to interact with GCN4 motifs (TGA(G/C)TCA) to regulate seed storage proteins (SSPs). In addition, OsSMF1 (also known as OsbZIP58) functions as a key regulator of starch synthesis in the rice seed. Quadruple 9-mer-based PBM (Q9-PBM) and electrophoretic mobility shift assay (EMSA) experiments revealed that OsSMF1 binds to the ACGT (CCACGT(C/G)), GCN4 (TGA(G/C)TCA), and GCN4-like (GGATGAC) motifs with Kd values of 0.3353 μM, 0.6458 μM, and 1.117 μM, respectively. We also identified 60 putative OsSMF1 target genes using a combination of data from expression microarrays and RiceArrayNet (RAN) analysis. Of these OsSMF1 target genes, 20, 22, and 17 genes contained ACGT, GCN4, and GCN4-like motifs within the 2-kb promoter region, respectively. In addition to known target genes, we also identified 35 potential OsSMF1 target genes that have not been previously described in immature seeds. We also confirmed that OsSMF1 directly regulates Os03g0168500 (thioredoxin-related protein), RPBF, NAC6, and two hypothetical proteins (Os12g0621600 and Os11g0582400) in vivo. This study suggests that OsSMF1 functions in a wide range of seed development processes with specific binding affinities for three DNA binding motifs
Drought and salinity are two major environmental factors determining plant productivity that due to their high magnitude of impact and wide distribution. The regulatory circuits include stress sensors, signaling pathways comprising a network of protein-protein reactions, transcription factors and promoters, and finally the output proteins or metabolites. Plant receptor-like kinases (RLKs) are transmembrane proteins family, are predicted to be major components of the signaling pathways that allow plants to respond to diverse environmental and development condition. Subfamily of Catharanthus roseus RLK1-like kinases (CrRLK1Ls) is a novel type of RLK, was identified in Arabidopsis with 17 members carrying a putative extracellular carbonhydrate-binding malectin-like domain. To study the function of CrRLK1Ls subfamily in rice which is a most widely consumed staple food, we produced the phylogenomic data with the integration of microarray-based anatomical and stress expression profiling data to the context of rice CrRLK1Ls family phylogenic tree. The expression profiling data are based on a large number of public microarray data such as 1150 Affymetrix arrays and 209 Agilent 44K arrays. Chromosomal localization of CrRLK1Ls reveals that three of 16 genes were tandem duplicated. Subsequently, we identified 7 genes that showed circadian regulation pattern and three genes of them simultaneously response to drought stress: two were down-regulated and one was up-regulated. Functional gene network development mediated by these stress responsible genes might be an useful foundation to explain the molecular mechanism of stress response mediated by this gene family.
Drought and salinity are two major environmental factors determining plant productivity that due to their high magnitude of impact and wide distribution. The regulatory circuits include stress sensors, signaling pathways comprising a network of protein-protein reactions, transcription factors and promoters, and finally the output proteins or metabolites. Plant receptor-like kinases (RLKs) are transmembrane proteins family, are predicted to be major components of the signaling pathways that allow plants to respond to diverse environmental and development condition. Subfamily of Catharanthus roseus RLK1-like kinases (CrRLK1Ls) is a novel type of RLK, was identified in Arabidopsis with 17 members carrying a putative extracellular carbonhydrate-binding malectin-like domain. To study the function of CrRLK1Ls subfamily in rice which is a most widely consumed staple food, we produced the phylogenomic data with the integration of microarray-based anatomical and stress expression profiling data to the context of rice CrRLK1Ls family phylogenic tree. The expression profiling data are based on a large number of public microarray data such as 1150 Affymetrix arrays and 209 Agilent 44K arrays. Chromosomal localization of CrRLK1Ls reveals that three of 16 genes were tandem duplicated. Subsequently, we identified 7 genes that showed circadian regulation pattern and three genes of them simultaneously response to drought stress: two were downregulated and one was up-regulated. Functional gene network development mediated by these stress responsible genes might be an useful foundation to explain the molecular mechanism of stress response mediated by this gene family.
Miniature inverted- repeat transposable elements are expected to play vital role in evolution of genes and genome of major eukaryotic organisms. However, there have been little reports on MITEs in B. rapa, a polyploidy model genome. We identified 13 novel MITE families in B. rapa genome by computational approach. Out of 13 MITEs families three, eight and two were classified under stowaway-like, tourist-like and hAT super families based on their unique structural characteristics. We characterized the members of 13 MITE families from the available 256 Mbp from whole genome draft sequences of B. rapa. We found ech MITE has high copiy number ranges from 14 to 977 which are distributed randomly along all the chromosomes. We also found more than 40% of the MITE members were associated with genes and gene rich regions. Furthermore, the polymorphism due to insertion and non-insertion of MITEs analysis suggest that MITEs are active in the genome. As, such the newly identified MITEs will provide a foundation for the further analysis of roles of MITEs in gene and genome evolution.