The desulfurizers facility is cylindrical shape. To operate properly it needs nozzles cleaned, get rid of lime adhesion and sludge, repair the wear and corrosion of facility regularly. For this purpose, workers shall access the ceiling or vertical wall at high place. Ordinary scaffoldings such as steel pipe scaffolding or system scaffolding have been using so that workers can access them. With these ordinary scaffoldings, openings around cylindrical wall are inevitable which make workers can expose always to the risk of falling. The purpose of this study is to develop customized scaffolding to minimize the openings to prevent workers form falling during maintenance it. It consists of a hexagonal central tower and six trapezoidal outer towers. And the bracing among the towers have connected each other for self-standing and for maintaining the structure of towers. Span decks, the circumference footstools, steps, etc. are laid on each floor. The safety is reviewed by structural analysis and performance test. With this study, openings each floor of this scaffold are removed. The gap between the cylindrical wall and the edge of the work stage is approximately 100 mm. Therefore, we expect that workers can work safely and efficiently.

Efficient infiltration of water through cell membranes is arbitrated by a family of transmembrane water channels called aquaporins (AQPs). Aquaporin belongs to a highly conserved group of membrane proteins called major intrinsic proteins that facilitate the transport of water and a variety of low molecular weight solutes across biological membranes,which is essential for plants to survive in stress conditions. This study identified 59 BrAQP genes from B. rapa database and Br135K microarray dataset, which was formed by applying low-temperature stresses to contrasting Chinese cabbage two inbreed lines, Chiifu and Kenshin. Based on phylogenetic analyses of BrAQPs revealed four distinct subfamilies, such as plasma membrane intrinsic proteins (PIP), tonoplast intrinsic proteins (TIP), NOD26-like intrinsic proteins (NIP), small basic intrinsic proteins (SIP) with aquaporin of Tomato and Arabidopsis thaliana. All BrAQP genes were firstly examined through homology study with existing biotic and abiotic stress resistance-related aquaporin genes of other plant species and found a high degree of homology. We selected PIP subfamily genes for expression analysis based on microarray data with high and differential transcript abundance levels and homology study with stress related aquaporin genes of other plant species. In our study, we characterized all B. rapa aquaporin genes and understanding the BrPIP subfamily gene function in plants under various environmental stimuli, the expressions of BrPIP genes under various abiotic stress conditions including cold, drought, salinity, water logging, ABA treatment and Fusarium oxysporum f. sp. Conglutinans infection were investigated by a quantitative real-time reverse transcription-PCR analysis. In our expression analysis, 4 BrPIP genes showed responsive expression against F. oxysporum f. sp. Conglutinans infection. The selected genes showed an organ-specific expression, and 12 out of 22 BrPIP genes were differentially expressed in Chiifu compared to Kenshin under cold stresses. Only 7 genes showed up regulation under drought stress and incase of salt stress 17 BrPIP genes were more responsiveness. Additionally, 18 BrPIP genes were up regulated by ABA treatment and all BrPIP genes showed down regulation under water logging stress. Together with expression and bioinformatic analyses, our results provides novel basis to allocate the stress-related biological function to each PIP gene.

Anthocyanins are responsible for vivid colors of flowers, fruits and vegetative tissues and biosynthesis of it is primarily controlled by several structural and regulatory genes. The regulatory mechanism of this pathway is still unknown. This study identified 19 transcription factors of Brassica rapa and investigated their regulatory function in anthocyanin biosynthesis pathway genes and cold and/or freezing tolerance in B. rapa. Expression analysis of these genes in the pigmented and non-pigmented portion of leaves of different lines of B. rapa revealed that BrMYB2-2 and BrTT8 showed responses contrasting with anthocyanin accumulation and cold stress. Sequences of these genes were analyzed and compared with similar gene sequences from other species and a high degree of homology with their respective functions was found. Co-regulated cis -elements were found in promoters of BrPAL1, BrCHS, BrF3H1, BrF3’H1, BrFLS, BrBAN, BrDFR8, BrANS1, and BrMYB2-2 and BrTT8 had binding sites of the promoters of those structural genes. Thus, the above results suggest the association of BrMYB2-2 and BrTT8 with regulation of anthocyanin biosynthesis pathway genes and cold and freezing stress tolerance and might be useful resources for development of cold resistant Brassica crops with desirable colors as well.

Bulb onion (Allium cepa) is one of the second most widely cultivated and consumed vegetable crops in the world. During winter where the temperature can be as low, plant could get cold injury and limit the production of bulb onion. However, the genomic resources available for bulb onion are still very limited. To date, no studies about heritably durable cold and freezing tolerance were carried out in bulb onion genotypes using high-throughput sequencing technology was applied. We sequenced cold (2°C) freezing (-5 and -15°C) treated and control (25°C) samples of contrasting genotypes of A. cepa lines and obtained 4,52,194,370 total high quality reads. After de novo assembly reads were assembled into 54,047 genes finally generated with an average length of 1,331 bp. Based on the similarity search aligning all genes with known public non-redundant (NR) database, including Swiss-prot, KEGG and COG. Differentially expressed genes (DEGs) were investigated using FPKM method. Overall, 92,862 genes were differentially regulated in all libraries were identified. Additionally, increase our understanding of the DEGs, we performed GO and KEGG pathway enrichment analyses. Based on FDR<=0.01 value in cold freezing tolerant line candidate genes were selected and discussed. Finally 25 candidate genes were examined using qRT-PCR were differentially regulated and known to be associated with cold and freezing stresses. Moreover, in silico prediction of putative molecular marker 4,437 SSRs and 6,076 SNPs. Our study is the first to provide the transcriptome sequence resource of Allium spp., for cold and freezing stress. We identified large set of genes to determine its DEGs profile under cold and freezing condition using two different genotypes. These data provides a valuable resource of genetic and genomic studies of Allium spp.

The TIFY family is composed of a plant-specific group of genes with diversity of functions. This family represents four subfamily of proteins viz. ZML, TIFY, PPD and JASMONATE ZIM-domain (JAZ) proteins. TIFY proteins especially, JAZ proteins have been reported to perform different biological processes, such as developmental and stresses and hormone responses in Arabidopsis and rice. However, there is no information about this family genes in Brassicaceae. This study identifies 36 TIFY genes in Brassica rapa, an economically important crop species from this family. An extensive in silico analysis through phylogenetic grouping, protein motif organization and intron-exon distribution also confirmed 4 subfamilies of BrTIFY proteins. Out of 35 BrTIFY genes, we identified 21 under JAZ subfamily besides 7 TIFY, 6 ZML and 2 PPD. An extensive expression profiling of 21 BrTIFY JAZs both in tissues and organs of B. rapa revealed differential expression patterns. Almost all the BrTIFY JAZs predominantly expressed in leaves and flower buds. Besides, in a flower stage specific expression analysis we observed 14 BrTIFY JAZs with constitutive expression patterns. This indicates BrTIFY proteins have a strong involvement in the development of B. rapa flowers. Our protein interaction study also reveals the strong association of these proteins with the fertility and defense processes of B. rapa. To elucidate the stress responsiveness of BrTIFY genes, we analyzed the low temperature-treated whole-genome microarray data set and found almost all the BrTIFY JAZs were having variable transcript abundance in two contrasting inbred lines of B. rapa. Subsequently, all 21 BrTIFY JAZs were validated in response to cold stress in the same two lines via qPCR, where 9 genes were found to show up- regulation. And, a high and differential qPCR expression pattern of all the BrTIFY JAZs was also recorded against JA. Additionally, BrTIFY JAZs were tested against salt, drought, Fusarium, ABA and SA treatments and a considerable number of genes were found to be induced. The extensive annotation and transcriptome profiling reported in this study will be useful for understanding the involvement of TIFY genes in stress resistance and different developmental functions, which ultimately provides the basis for functional characterization and exploitation of the candidate genes for genetic engineering of B. rapa.

Onion and other Allium vegetables have been valued since antiquity for their pungent flavor and aroma. Modern science has confirmed traditional benefits that the organosulfur compounds that impart flavor also confer significant human health benefits such as reduced blood clotting and antimicrobial properties. Glucose, fructose and sucrose comprises majority of onion bulb dry matter content. The sugars, pyruvic acid accumulation and transcript level of some transcription factors involved in the biosynthesis of high sugars and pyruvic acid. These profiles were compared with two different lines 36101 (early) and 36122(Late) of bulb onion (Allium cepa L.) growing under drought and photoperiod condition using High Performance Liquid Chromatography (HPLC) and Quantitative real time PCR using FT genes. We identified the gene AcFT4 was responsible for early and late bulb intiation in the onion lines. The cultivar lines 36101and 36122 were used to identify potential genes controlling pungency and sugar. The comparative analysis of two lines showed significant positive phenotypic and genetic correlations. Sugar and pungency profile showed significant difference between two lines. FT gene expression and pungency level was high in onion lines during drought stress. In this study, we proposed the biochemical characterization of two line and genes involved in the bulb formation were also studied. There is a correlation between sugars and pungency level during the drought stress. These results could be presumably used as useful information to obtain onion varieties rich in sugars and pungency.

Blackleg disease caused by Leptosphaeria maculans, is the most devastating disease of Brassica germplam worldwide that causes million tonnes of crop losses per year throughout the world. To date, a total of 12 race-specific resistance genes of Brassica napus to L. maculans have been reported but linkage mapping analysis reveals that all of those loci are located in A genome i.e., in B. rapa chromosomes. B. oleracea has high ancestral synteny with B. rapa through their evolution. We believe that presence of qualitative resistance is possible in B. oleracea germplasm. The present study was therefore planned to find out any race-specific qualitative resistance gene present in C genome of B. oleracea. A total of 16 microsatellite markers were used which are linked to seven different Rlm and Lep genes of B. napus to screen 32 inbred lines of cabbage. Primers were designed based on homology assessment in corresponding nucleotide sequence available in Bolbase (a B. oleracea genome database, http://www.ocri-genomics.org/bolbase/index.html), located in B. oleracea scaffolds/chromosomes. Out of 16 SSR markers, 13 were found polymorphic which indicates possible existence of resistant genes in cabbage lines. The inbred lines are then assessed against two L. maculans stains with known avirulent genes. Some inbred lines were hypersensitive against gene-specific virulent strains of L. maculans that confirmed existence of Rlm1, Rlm2, Rlm4, LepR3 and LepR4 in the cabbage lines. In this way we were able to select out resistant and susceptible lines against each resistant gene. The gene-specific polymorphic SSR marker regions were cloned and sequenced and candidate SNPs were identified for confirmation of their functionality.

The Alfin-like transcription factor family is one of the important gene families in eukaryotic plants. They are involved in many biological processes, such as lignocellulosic wall biosynthesis, meristem development, metabolite transport, and responses to biotic and abiotic stresses. But the regulatory mechanism of these genes involved in stresses responses is still unrevealed. In this study, we identified a total of 16 Alfin-like genes from Brassica rapa database. The 16 putative Alfin-like proteins were divided into four groups (group I-IV) based on structural and phylogenetic analyses. Accordingly, this study analyzed stress resistance-related functions of all B. rapa Alfin-like (BrAL) genes through a homology study with existing biotic and abiotic stress resistance-related Alfin-like genes of other plant species and found a high degree of similarity with them. Subsequently, these genes were further investigated by real-time quantative PCR under cold, salt and drought stresses and after infection with Fusarium oxysporum f. sp. conglutinans in B. rapa. These genes showed an organ specific expression and all genes differentially expressed in Chiifu compared to Kenshin under cold stress. Ten and seven BrALs responded highly in Kenshin compared to Chiifu under salt and drought stresses respectively. In addition, six BrAL genes showed responsive expression after Fusarium oxysporum f. sp. conglutinans infection in B. rapa. Interestingly, four BrAL genes showed responses against both biotic and abiotic stress factors. Thus, our result provides a useful reference data set as the basis for functional analysis and utilization in the resistance molecular breeding of B. rapa.

Heterosis is very important for breeding hybrid cultivars and is intensively used to increase the productivity of crop plants. But the molecular basis of heterosis is still unrevealed to the scientists. This study selected 51 heterosis associated genes of Arabidopsis of different family on the basis of their high differential expression in a hybrid compared to its midparent value and identified their orthologues in Brassica oleracea. Then the selected B. oleracea genes were characterized based on their structural properties, recognized functions and expression patterns in a cabbage hybrid progeny (Cabbage-36) of crosses between Cabbage-34 and Cabbage-35 accessions. Among these genes, a good number were found to express highly in the hybrid then the midparent value and better parent in some cases. Moreover, these highly expressed genes are mostly related to the yield contributing characters. Cotyledon and young leaf sizes of these three genotypes also well correlated with gene expression. Thus, it can be said that, the identified genes might be associated with the mechanism of heterosis of B. oleracea hybrid and provide a foundation for the exploration of gene regulatory networks associated with the specification of the phenomenon heterosis in the plant life cycle. Subsequently, these genes would be useful resources for molecular hybrid breeding in Brassica crops as well.

Flavonoids are divided into several structural classes, including anthocyanins, which provide flower and leaf colors and other derivatives with diverse roles in plant development and interactions with the environment. This study characterized four Anthocyanidin Synthase (ANS) genes of Brassica rapa, a structural gene of anthocyanin biosynthetic pathway, and investigated their association with cold and freezing tolerance in B. rapa. Sequences of these genes were analyzed and compared with similar types of gene sequences of other species and found a high degree of homology with their respective functions. In the organ specific expression analysis, these genes showed expression only in the colored portion of leaves of different lines of B. rapa. On the other hand, BrANS genes also showed differential expression with certain time course of cold stress treatment in B. rapa. Thus, the above results suggest probable association of these genes with anthocyanin biosynthesis and cold and freezing tolerance and might be useful resources for developing cold resistant Brassica crops with desirable colors as well. The present work may help explore the molecular mechanism that regulates anthocyanin biosynthesis and its response to abiotic stress at the transcriptional level in plants.

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.

Cabbage (Brassica oleracea) is one of the most important vegetable crops in the world. Yet, its sensitivity to cold stress, especially at the seedling stage, could limit the production. Until now, only, few studies about heritably durable cold tolerance were carried out in cabbage. Hence this study was done to characterize the transcriptome profiles of two cabbage genotypes with contrasting responses to cold stress using Illumina Hiseq short read (paired-end) sequencing technology. MicroRNAs (miRNAs) represent a class of short, non-coding, endogenous RNAs which play important roles in post-transcriptional regulation of gene expression. Thisstudy,wesoughttoprovideamorecomprehensivepredictionofB. oleracea cold responsive miRNAs based on high through put sequencing using two contrasting genotypes. The raw sequences were processed for removal of poor-quality and adaptor sequences. Then, the high quality unigenes (58,094) reads were applied for length filtering. Then, unigenes reads were used in a BLASTN search against of Rfam database and known miRNA database (miRBase 18.0) to removal of non-coding RNA’s and identifies conserved miRNA’s in B. oleracea. Further, novel reads were searched against B. oleracea genome. Their flanking sequences in the genome were used to predict their secondary structures, target prediction, and functional analysis. This is first report to identify novel miRNAs for cold stress through high throughput techniques. Our findings will provide an overview of potential miRNAs involved in cold stress, which may provide important clues on the function of miRNAs in from B. oleracea and other closely related Brassica species.

MADS-box transcription factor (TF), primarily involved in the floral organ specification with other several aspects of plant growth and development. Whole genome survey of B. rapa revealed 167 MADS-box genes and categorized into MIKCc, MIKC*, Mα, Mβ and Mγ groups based on phylogeny, protein motif structure and exon-intron organizations. MIKCc group belongs 89 genes, which is the highest in number than in any other crops till date. The MIKCc group has further classified into 13 sub-families. In case of chromosomal localization, remarkably 57 MIKCc type MADS-box genes were found in the duplicated segments of B. rapa genome, whereas only 4 M-type genes have resulted from tandem duplications. Besides floral and vegetative tissue expression we also identified MADS-box genes with their male and female gametophyte specific expression in different stages of flower bud development. Furthermore, from a low temperature treated whole genome microarray data set 19 BrMADS genes were found to show variable transcript abundance in two contrasting double haploid lines of B. rapa. Subsequently, the responsive genes were investigated under three abiotic stresses where they showed differential and corresponsive expression patterns. An extensive annotation and transcriptome profiling undertaken in this study might be useful for understanding the involvement of MADS-box genes in stress resistance besides their growth and developmental functions, which ultimately will provide the basis for functional characterization and exploitation of the candidate genes in the genetic engineering study of B. rapa

Flavonoids including anthocyanins provide flower and leaf colors and other derivatives that play diverse roles in plant development and interactions with the environment and dihydroflavonol 4-reductase (DFR) is part of an important step in the flavonoid biosynthesis pathway of anthocyanins. This study characterized 12 DFR genes of Brassica rapa and investigated their association with anthocyanin coloration, cold and freezing tolerance in several genotypes of B. rapa. Sequences of these genes were analyzed and compared with DFR gene sequences from other species and a high degree of homology was found. Constitutive expression of them in several pigmented and non-pigmented lines of B. rapa showed a correlation with anthocyanin accumulation only for BrDFR8 and 9. Conversely, BrDFR genes also showed responses to cold and freezing stress treatment in B. rapa. BrDFRs were also shown to be regulated by two transcription factors, BrMYB2-2 and BrTT8, contrasting with anthocyanin accumulation and cold and freezing stress. Thus, the above results suggest the association of these genes with anthocyanin biosynthesis and cold and freezing stress tolerance and might be useful resources for development cold and/or freezing resistant Brassica crops with desirable colors as well. The findings presented here may also help explore the molecular mechanism that regulates anthocyanin biosynthesis and its response to abiotic stress at the transcriptional level in plants.

Numerous environmental stresses, such as abiotic and biotic stresses, cause significant yield loss in crops and can significantly affect their development. Un the field conditions, crops are exposed to a variety of concurrent stresses. Combined high temperature and linked diseases can cause considerable damage that eventually leads to crop death. Hence, this study was conducted to characterize the genes encoding the nucleotide-binding site (NBS) motif obtained from transcriptome profiles of two cabbage genotypes with contrasting responses to heat stress. We selected 80 up-regulated genes form a total of 264 loci, among which 17 were confirmed to be complete and incomplete members of the TIR-NBS-LRR (TNL) class families, and another identified as a NFYA-HAP2 family member. Expression analysis using qRT-PCR revealed that 8 genes showed significant responses to heat shock treatment and F. oxysporum infection. Additionally, in the commercial B. oleracea cultivars with resistance to F. oxysporum, Bol007132, Bol016084, and Bol030522 genes showed dramatically higher expression levels in the F. oxysporum resistant line than the intermediate and susceptible lines. The results of this study may facilitate the identification and development of molecular markers based on multiple stress resistance genes related to heat and fungal stress under field conditions in B. oleracea.

Calcium-binding proteins, like calcineurin B-like (CBL) proteins, represent important roles in plant calcium signaling. Calcium signals mediate a multitude of plant responses to external stimuli and regulate a wide range of physiological processes including pathogens, abiotic stresses and hormones. These proteins form a complex network with their target kinases being the CBL-interacting protein kinases (CIPKs). CBL genes play vital roles in multiple abiotic stress response pathways whereas some of these are more specifically involved in mediating ABA signaling. In this study, we collected 17 CBL genes designated as B. rapa CBL (BrCBL) from the Brassica database and analyzed the sequences. In comparison analysis, these genes showed high homology with published CBL genes of other species. An organ specific expression of these genes was observed in different organs of chinese cabbage plants. In addition, six BrCBL genes showed responsive expression after cold and drought stress treatments at certain time courses. All these data revealed that these CBL genes might be useful resources in developing abiotic stresses resistance Brassica.