Molecular markers, such as PCR-based and SNP-based markers, are extremely useful for plant genetics and crop breeding. Marker-assisted selection (MAS) has been widely applied in plant breeding to improve crop yield, quality, and tolerance to biotic and abiotic stresses. To develop gene-based (or -specific) molecular markers, three different approaches have been used in Brassica species: Known-gene-based, RNA seq/Exon-based and RNA seq/Intron-based molecular marker development for several years. Using these techniques, molecular markers have been developed to identify flowering time, anthocyanin accumuation and abiotic stresses in B. rapa and B. oleracea. Markers were distributed in exons as well as introns, and coding sequences and untranslated regions (UTRs). All markers developed have been transformed into SNP marker after HRM confirmation. I will discuss efficiency, accuracy, and potential problems and contribution of these markers for Brassica breeding.

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.

Fusarium crown root rot (FCRR) is a severe fungal disease caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL) in tomato. Resistance to FORL is conferred by single dominant locus Frl on chromosome 9, but its precise genomic location is not clearly determined. In this study, detailed location of Frl was assessed by using a set of molecular markers physically anchored on Chr.9 and F2 and RIL population derived from FORL-resistant inbred AV107-4 (S.lycopersicum) x susceptible L3708 (S. pimpinellifolium). Bioassay of the two populations with a FORL strain isolated from Korea resulted in single dominant heritance of the resistance. Two SCAR and 11 CAPS markers encompassing 3.6Mb~72Mb of Chr.9 were developed from the Tomato-EXPEN 2000 map and SolCAP SNP-array analysis. These markers were genotyped on 345 F2 plants. A high level of cosegregation with the resistance were observed for 5 markers which were mapped at a large physical interval of 5.1Mb (T1212) to 46.4Mb (SSR237), indicating that genetic recombination was highly suppressed in this region. Cosegregation of these markers with Frl was confirmed by using 126 RILs. The results implied that, in contrast with the previously reported long arm, Frl is present on a pericentromeric region of short arm of Chr. 9, in which crossing-over is severely suppressed. The marker set was further tested on 12 FORL-resistance or susceptibility commercial cultivars. Unlike the biparental populations, frequent linkage break was observed for T1212 and D4 in commercial cultivars. T1212 and D4 showed 50% and 100% match with the phenotype, respectively. D4, a CAPS, was converted to a high resolution melting (HRM) marker and tested on 55 breeding lines from private seed companies (Fig.3). All breeding lines showed the HRM genotype for resistance allele, indicating that D4 can be useful for selecting FORL-resistance tomato plants.

Clubroot is a devastating disease caused by Plasmodiophora brassicae and results in severe losses of yield and quality in Brassica crops including Brassica oleracea. Therefore, it is important to identify resistance gene for CR disease and apply it to breeding of Brassica crops. In this study, we applied genotyping-by-sequencing (GBS) technique to construct high resolution genetic map and mapping of clubroot resistance (CR) genes. A total of 18,187 GBS markers were identified between two parent lines resistant and susceptible to the disease, of which 4,103 markers were genotyped in all 78 F2 plants generated from crossing of both parent lines. The markers were clustered into nine linkage groups spanning 879.9 cM, generating high resolution genetic map enough to refine reported reference genome of cabbage. In addition, through QTL analysis using 78 F2:3 progenies and mapping based on the genetic map, two and single major QTLs were identified for resistance of race 2 and race 9 of P. brassicae, respectively. These QTLs did not show collinearity with CR loci found in Chinese cabbage (Brassica rapa) but roughly overlapped with CR loci identified in cabbage for resistance to race 4. Taken together, genetic map and QTLs obtained in this study will provide valuable information to improve reference genome and clubroot resistance in cabbage.

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.

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.

Plant bZIP transcription factors play crucial roles in biological processes. In this study, 136 putative bZIP transcription members were identified in Brassica rapa. The bZIP family can be divided into nine groups according to the specific amino acid rich domain in Brassica rapa. To screen the cold stress responsive BrbZIP genes, we evaluated whether the transcription patterns of the BrbZIP genes were enhanced by cold treatment in the inbred lines, Chiifu and Kenshin, by microarray data analysis and qRT-PCR. The expression level of six genes increased significantly in Kenshin, but these genes were unchanged in Chiffu. Additionally, homo- and hetero-dimerization test between selected bZIP proteins indicated the Bra020735 is a key regulator in cold response. These findings suggest that the six genes that encoded proteins containing N-rich regions might be involved in cold stress response. These results presented herein provide valuable information regarding the molecular basis of the bZIP transcription factors and their potential function in regulation growth and development, particularly in cold stress response.

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.

Mitochondria are essential organelles of eukaryotic cells and plant cells contain varying numbers of mitochondrial genome sequences. Sizes and shapes of mitochondria differ within a tissue or in the same cells. Previously sequenced complete mitochondrial genome (NC_016118) of Brassica oleracea size was 360,271 bp, where segmental duplication (repeat block) was 141,800 bp. In this study, we resequenced this whole mitochondrial genome by using WGS (whole genome sequencing) and assembled organelles genome method (unpublished). Newly sequenced mitochondrial genome length was 219,975 bp and circle form. A new sequence segment of approximately 4,800 bp was obtained compared to the previous genome sequence without any large repeat block. Newly obtained mitochondria genome sequence was compared with recently reported mitochondria genome sequences of various species (B. oleracea, B. juncea, B. rapa, B. napus and B. carinata) and subspecies (cabbage, cauliflower, brussels sprouts, kohlrabi, broccoli and kale) by PCR using primers specifying different region of genome sequences. PCR analysis results have also confirmed the variation between previous and newly sequenced mitochondrial genome circles form. Thus, the results suggest new B. oleracea mitotype, including evolutionary events such as inheritance, rearrangement, genome compaction, and diversity

Bulb onion (Allium cepa), which belongs to the family Amaryllidaceae, is one of the oldest vegetative crops known to humans. Despite its high economic value, only a few reports are available on the use of molecular markers in genetic diversity analysis of Allium cepa for its improvement. Molecular genetic markers have been widely used as powerful tools for analyzing the plant genome. In particular, Microsatellites or simple sequence repeats (SSRs) markers are tandem repeats of one to six bp in length and have been proven to be the most powerful polymerase chain reaction (PCR)-based DNA markers in plant diversity analysis. In this study, the genomic DNA was isolated from different Allium cepa lines. The ESTs and gDNA sequences of onion were collected from National Center for Biotechnology information. The SSRs with two to five motifs over a length of 12 bp, were identified using SSRIT (Gramene) software. The PCR products of 100 to 350 bp in length containing SSRs, primers was designed using Primer3 with lengths of 20 to 24 bp and a melting temperature of 60℃. The SSR markers with high polymorphism-information content (PIC) levels was useful for collecting progeny with high genetic homogeneity for onion breeding, and to obtain representative marker sets for genetic tests. The SSR Finder program and the developed SSR markers could be a useful resource for genetic diversity and purity testing in onion.

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

Onion is one of the most widely consumed vegetables. There are many cultivars, which are grouped according to skin color as yellow, white or red. Onions can also be classified as sweet or non-sweet. Their importance in cooking comes from their typical taste and flavour. The sugars, pyruvic acid accumulation and transcript level of some transcription factors involved in the biosynthesis of high sugars and pyruvic acid was analyzed at different stages of bulb onion (Allium cepa) growing under light and dark condition using High Performance Liquid Chromatography (HPLC) and Quantitative real time PCR. A genetic map and cultivar lines 36101and 36122 were used to identify transcription factors controlling pungency and sugar. We compared 2 different lines for low pungency and high sugars during water and photoperiod stress, which showed significant positive phenotypic and genetic correlations. These results could be presumably used as useful information to obtain onion varieties rich in sugars.

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.