Chinese Cabbage Inbred Line Research Articles

Long noncoding RNAs in Brassica rapa L. following vernalization

Brassica rapa L. is an important agricultural crop that requires a period of prolonged cold for flowering. This process is known as vernalization. Studies have shown that long noncoding RNAs (lncRNAs) play important roles in abiotic stress responses and several cold-responsive noncoding RNAs have been suggested to be involved in vernalization. We examined the transcriptome of the Chinese cabbage inbred line (B. rapa L. var. pekinensis) RJKB-T24, and identified 1,444 long intergenic noncoding RNAs (lincRNAs), 551 natural antisense transcripts (NATs), and 93 intronic noncoding RNAs (incRNAs); 549 of the 2,088 lncRNAs significantly altered their expression in response to four weeks of cold treatment. Most differentially expressed lncRNAs did not lead to a change of expression levels in mRNAs covering or near lncRNAs, suggesting that the transcriptional responses to four weeks of cold treatment in lncRNA and mRNA are independent. However, some differentially expressed mRNAs had NATs with expression altered in the same direction. These genes were categorized as having an abiotic stress response, suggesting that the paired-expression may play a role in the transcriptional response to vernalization or cold treatment. We also identified short-term cold treatment induced NATs in BrFLC and BrMAF genes, which are involved in vernalization. The lncRNAs we identified differed from those reported in Arabidopsis thaliana, suggesting the role of lncRNAs in vernalization differ between these two species.

Identification and development of a core set of informative genic SNP markers for assaying genetic diversity in Chinese cabbage

Rapid, economical, and reliable genotyping is an important requirement for germplasm analysis and cultivar identification in crop species. Chinese cabbage (Brassica rapa L. subsp. pekinensis (Lour.) Hanelt) originated in China and is now an economically important vegetable crop worldwide, especially in East Asia. In this study, we evaluated 1167 single nucleotide polymorphisms (SNPs) among 166 representative Chinese cabbage inbred lines using a KASP genotyping assay. On the basis of polymorphisms and principal component analysis, we selected 60 core SNPs distributed on all Brassica rapa chromosomes with allele frequencies sufficiently balanced so as to provide adequate information for genetic identification. The core set of SNPs was used for construction of a neighbor-joining dendrogram, in which the 166 inbred lines were clustered into spring, summer, and autumn ecotype groups. Clustering of the ecotype groups was better resolved than that achieved with 1167 and 360 polymorphic SNP datasets. Stability and resolution of the core SNP markers were tested using 178 commercial hybrid Chinese cabbage cultivars to confirm their utility in genetic identification. The set of 60 informative and stable SNP markers showed high discriminatory power and relatively uniform genomic distribution (4–9 markers per chromosome). The SNPs represent a cost-efficient and accurate marker set for germplasm analysis and cultivar identification and are suitable for molecular marker-assisted breeding in Chinese cabbage.

Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino

AbstractSingle‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.

Characterization of Non-heading Mutation in Heading Chinese Cabbage (Brassica rapa L. ssp. pekinensis).

Heading is a key agronomic trait of Chinese cabbage. A non-heading mutant with flat growth of heading leaves (fg-1) was isolated from an EMS-induced mutant population of the heading Chinese cabbage inbred line A03. In fg-1 mutant plants, the heading leaves are flat similar to rosette leaves. The epidermal cells on the adaxial surface of these leaves are significantly smaller, while those on the abaxial surface are much larger than in A03 plants. The segregation of the heading phenotype in the F2 and BC1 population suggests that the mutant trait is controlled by a pair of recessive alleles. Phytohormone analysis at the early heading stage showed significant decreases in IAA, ABA, JA and SA, with increases in methyl IAA and trans-Zeatin levels, suggesting they may coordinate leaf adaxial-abaxial polarity, development and morphology in fg-1. RNA-sequencing analysis at the early heading stage showed a decrease in expression levels of several auxin transport (BrAUX1, BrLAXs, and BrPINs) and responsive genes. Transcript levels of important ABA responsive genes, including BrABF3, were up-regulated in mid-leaf sections suggesting that both auxin and ABA signaling pathways play important roles in regulating leaf heading. In addition, a significant reduction in BrIAMT1 transcripts in fg-1 might contribute to leaf epinastic growth. The expression profiles of 19 genes with known roles in leaf polarity were significantly different in fg-1 leaves compared to wild type, suggesting that these genes might also regulate leaf heading in Chinese cabbage. In conclusion, leaf heading in Chinese cabbage is controlled through a complex network of hormone signaling and abaxial-adaxial patterning pathways. These findings increase our understanding of the molecular basis of head formation in Chinese cabbage.

Genome-wide characterization of DNA methylation, small RNA expression, and histone H3 lysine nine di-methylation in Brassica rapa L.

Epigenetic gene regulation is crucial to plant life and can involve dynamic interactions between various histone modifications, DNA methylation, and small RNAs. Detailed analysis of epigenome information is anticipated to reveal how the DNA sequence of the genome is translated into the plant’s phenotype. The aim of this study was to map the DNA methylation state at the whole genome level and to clarify the relationship between DNA methylation and transcription, small RNA expression, and histone H3 lysine 9 di-methylation (H3K9me2) in Brassica rapa. We performed whole genome bisulfite sequencing, small RNA sequencing, and chromatin immunoprecipitation sequencing using H3K9me2 antibody in a Chinese cabbage inbred line, RJKB-T24, and examined the impact of epigenetic states on transcription. Cytosine methylation in DNA was analysed in different sequence contexts (CG, CHG, and CHH) (where H could be A, C, or T) and position (promoter, exon, intron, terminator, interspersed repeat regions), and the H3K9me2 and 24 nucleotide small interfering RNAs (24 nt-siRNA) were overlaid onto the B. rapa reference genome. The epigenome was compared with that of Arabidopsis thaliana and the relationship between the position of DNA methylation and gene expression, and the involvement of 24 nt siRNAs and H3K9me2 are discussed.

Identification and Mapping of the Clubroot Resistance Gene CRd in Chinese Cabbage (Brassica rapa ssp. pekinensis).

The rapid spread of clubroot disease, which is caused by Plasmodiophora brassicae, threatens Brassicaceae crop production worldwide. Breeding plants that have broad-spectrum disease resistance is one of the best ways to prevent clubroot. In the present study, eight Chinese cabbage germplasms were screened using published clubroot-resistant (CR) loci-/gene-linked markers. A CR gene Crr3 potential carrier “85-74” was detected which linked to marker BRSTS61; however, “85-74” shows different responses to local pathogens “LAB-19,” “LNND-2,” and “LAB-10” from “CR-73” which harbors Crr3. We used a next-generation sequencing-based bulked segregant analysis approach combined with genetic mapping to detect CR genes in an F2 segregant population generated from a cross between the Chinese cabbage inbred lines “85-74” (CR) and “BJN3-1” (clubroot susceptible). The “85-74” line showed resistance to a local pathogen “LAB-19” which was identified as race 4; a genetic analysis revealed that the resistance was conferred by a single dominant gene. The CR gene which we named CRd was mapped to a 60 kb (1 cM) region between markers yau389 and yau376 on chromosome A03. CRd is located upstream of Crr3 which was confirmed based on the physical positions of Crr3 linked markers. The identification of CRd linked markers can be applied to marker-assisted selection in the breeding of new CR cultivars of Chinese cabbage and other Brassica crops.

Identification of DNA methylated regions by using methylated DNA immunoprecipitation sequencing in Brassica rapa

DNA methylation is an epigenetic gene regulatory mechanism that plays an essential role in gene expression, transposon silencing, genome imprinting and plant development. We investigated the influence of DNA methylation on gene expression in Brassica rapa L., to understand whether epigenetic differences exist between inbred lines. Genome-wide DNA methylation was analysed by methylated DNA immunoprecipitation sequencing (MeDIP-seq) of 14-day-old first and second leaves from two inbred lines of Chinese cabbage, one susceptible and one resistant to fusarium yellows caused by Fusarium oxysporum f. sp. conglutinans. MACS (model-based analysis for ChIP-seq) identified DNA methylation peaks in genic regions including 2 kb upstream, exon, intron and 2 kb downstream. More than 65% of genes showed similar patterns of DNA methylation in the genic regions in the two inbred lines. DNA methylation states of the two inbred lines were compared with their transcriptome. Genes having DNA methylation in the intron and in the 200 bp upstream and downstream regions were associated with a lower expression level in both lines. A small number of genes showed a negative correlation between differences in DNA methylation levels and differences in transcriptional levels in the two inbred lines, suggesting that DNA methylation in these genes results in transcriptional suppression.

Establishment and application of molecular ID in the main inbred lines of Chinese cabbage.

Chinese cabbage is an important cruciferous vegetable in China. The differences in the morphology and other characteristics of the different varieties of Chinese cabbage are generally caused by their different genes. Using the simple sequence repeat (SSR) DNA molecular markers is an effective way to identify different genotypes. The identification of a genetic relationship is a key point in the breeding process, and it plays an important role in guiding parent selection and breeding of high-yield varieties. Moreover, the establishment of genomic fingerprints is significant for plant variety protection. Three to five SSR sites were selected from each of the 10 Chinese cabbage chromosomes on the basis of the abundance of SSR loci on them. According to the differences in the SSR polymorphic bands, a genomic fingerprint comprising 36 different loci was established in the 20 main inbred lines of Chinese cabbage, and this fingerprint was converted digitally into a molecular ID with 36 numbers based on the 36 SSR sites. The utility of this core set SSRs was demonstrated in 20 main inbred lines of Chinese cabbage, which could be placed into six clusters that were largely consistent with previous classification based on morphology data. Moreover, the molecular ID of an F1 hybrid can be deduced from its parents molecular IDs, and its purity can be determined by selecting one or two SSR loci from the 36 different loci.

Introgression of clubroot resistance into an elite pak choi inbred line through marker‐assisted introgression breeding

AbstractClubroot is a soilborne disease that severely infects cruciferous species. Pak choi (Brassica rapa subsp. chinensis) is an economically important cruciferous crop cultivated throughout the world. However, no clubroot‐resistant germplasms have been identified in pak choi to date. To improve disease resistance, we used marker‐assisted selection (MAS) to introgress the clubroot resistance (CR) trait from the ‘CCR13685’ Chinese cabbage (B. rapa subsp. pekinensis) inbred line into an elite pak choi inbred line, ‘GHQ11021’. Genetic analysis of F2 and BC1 progeny showed that CR of ‘CCR13685’ was controlled by a single dominant gene. We designed nine candidate sequence‐characterized amplified region markers, K‐1 to K‐9, based on two molecular markers linked to the CR gene. We found that K‐3 co‐segregated with CR and an inoculation test confirmed that K‐3 could be used for MAS. Two introgression lines, BC3‐1‐4 and BC3‐2‐18, were developed using K‐3 for foreground selection. These lines displayed the same phenotypic properties as ‘GHQ11021’, but were highly resistant to clubroot, indicating that the CR gene of ‘CCR13685’ had been successfully introduced into pak choi.

Characterization and expression profiling of MYB transcription factors against stresses and during male organ development in Chinese cabbage (Brassica rapa ssp. pekinensis)

MYB proteins comprise a large family of plant transcription factors that play regulatory roles in different biological processes such as plant development, metabolism, and defense responses. To gain insight into this gene superfamily and to elucidate its roles in stress resistance, we performed a comprehensive genome-wide identification, characterization, and expression analysis of MYB genes in Chinese cabbage (Brassica rapa ssp. pekinensis). We identified 475 Chinese cabbage MYB genes, among which most were from R2R3-MYB (256 genes) and MYB-related (202) subfamilies. Analysis of sequence characteristics, phylogenetic classification, and protein motif structures confirmed the existence of several categories (1R, 2R, 3R, 4R, and 5R) of Chinese cabbage MYB genes, which is comparable with MYB genes of other crops. An extensive in silico functional analysis, based on established functional properties of MYB genes from different crop species, revealed 11 and four functional clades within the Chinese cabbage R2R3-MYB and MYB-related subfamilies, respectively. In this study, we reported a MYB-like group within the MYB-related subfamily contains 77 MYB genes. Expression analysis using low temperature-treated whole-genome microarray data revealed variable transcript abundance of 1R/2R/3R/4R/5R-MYB genes in 11 clusters between two inbred lines of Chinese cabbage, Chiifu and Kenshin, which differ in cold tolerance. In further validation tests, we used qRT-PCR to examine the cold-responsive expression patterns of 27 BrMYB genes; surprisingly, the MYB-related genes were induced more highly than the R2R3-MYB genes. In addition, we identified 10 genes with corresponsive expression patterns from a set of salt-, drought-, ABA-, JA-, and SA-induced R2R3-MYB genes. We identified 11 R2R3-MYBs functioning in resistance against biotic stress, including 10 against Fusarium oxysporum f.sp. conglutinans and one against Pectobacterium carotovoram subsp. caratovorum. Furthermore, based on organ-specific expression data, we identified nine R2R3-MYBs that were constitutively expressed in male reproductive tissue, which may provide an important key for studying male sterility in Chinese cabbage. The extensive annotation and transcriptome profiling reported in this study will be useful for understanding the involvement of MYB genes in stress resistance in addition to their growth regulatory functions, ultimately providing the basis for functional characterization and exploitation of the candidate MYB genes for genetic engineering of Chinese cabbage.

Evaluation of Clubroot Resistance in Chinese Cabbage and Its Inheritance in the European Turnip Line ‘IT033820’, a New Genetic Resource

Clubroot caused by the protist Plasmodiophora Brassicae is one of the most destructive diseases of Brassica crops. Developing Chinese cabbage cultivars with durable clubroot resistance (CR) is an important goal of breeding programs, which will require new genetic resources to be identified and introduced. In this study, we evaluated resistance to P. Brassicae race 4 using 26 Chinese cabbage (B. rapa ssp. pekinensis.) cultivars compared to the clubroot-susceptible Chinese cabbage inbred line ‘BP079’ and the clubroot-resistant European turnip (B. rapa ssp. rapifera) inbred line ‘IT033820’. No symptoms of clubroot disease were found in ‘IT033820’ infected with P. Brassicae race 4, whereas the Chinese cabbage cultivars exhibited disease symptoms to various degrees. The Chinese cabbage cultivars that were reported to be clubroot-susceptible were susceptible to P. Brassicae race 4; however, seven of the 20 cultivars reported to be clubroot-resistant were susceptible to this race of P. Brassicae to varying degrees. Resting spores of P. Brassicae were abundant within the infected root tissues of ‘BP079’, as revealed by light microscopy and scanning electron microscopy (SEM), but they were not detected in root tissues of ‘IT033820’. Although resting spores were not detected by light microscopy in root tissues of the clubroot-resistant Chinese cabbage cultivar ‘Kigokoro 75’, a few spores were observed by SEM. The F1 hybrids from a cross between ‘IT033820’ and ‘BP079’ showed no disease symptoms, and all BC1P1 progenies from a cross between the F1 hybrid and ‘IT033820’ exhibited a resistance phenotype. In the BC1P2 population from a cross between the F1 hybrid and ‘BP079’, this trait segregated at a ratio of 3(R):1(S) (χ2 = 1.333, p = 0.248) at a 5% significance level. Inoculated BC1P2 plants were either highly resistant or highly susceptible to the pathogen, indicating that the CR to race 4 of P. Brassicae carried by ‘IT033820’ is dominant. In the F2 population, this trait segregated at a ratio of 15(R):1(S) (χ2 = 0.152, p = 0.696) at a 5% significance level, suggesting that CR in ‘IT033820’ is mainly controlled by two dominant genes. Therefore, ‘IT033820’ represents a promising genetic resource for developing durable CR breeding lines in Chinese cabbage.

Genetic characterization of inbred lines of Chinese cabbage by DNA markers; towards the application of DNA markers to breeding of F1 hybrid cultivars.

Chinese cabbage (Brassica rapa L. var. pekinensis) is an important vegetable in Asia, and most Japanese commercial cultivars of Chinese cabbage use an F1 hybrid seed production system. Self-incompatibility is successfully used for the production of F1 hybrid seeds in B. rapa vegetables to avoid contamination by non-hybrid seeds, and the strength of self-incompatibility is important for harvesting a highly pure F1 seeds. Prediction of agronomically important traits such as disease resistance based on DNA markers is useful. In this dataset, we identified the S haplotypes by DNA markers and evaluated the strength of self-incompatibility in Chinese cabbage inbred lines. The data described the predicted disease resistance to Fusarium yellows or clubroot in 22 Chinese cabbage inbred lines using gene associated or gene linked DNA markers.

Interspecific hybridisation of cytoplasmic male-sterile rapeseed with Ogura cytoplasm and Brassica rapa var. pekinensis as a method to obtain male-sterile Chinese cabbage inbred lines

AbstractDuring 2011–2014, three self-compatible Chinese cabbage inbred lines (Brassica rapa, genome AA 2n = 20) were sexually hybridised with cytoplasmic male-sterile (CMS Ogu-INRA) and fertile lines of rapeseed (Brassica napus var. oleifera, genome AACC 2n = 38). Interspecific F1 hybrids and their consecutive back-crosses (BCs) toward B. rapa were obtained, and their morphological and commercial characteristics, as well as their ability for generative propagation, were analysed. Cytometric and morphological analyses confirmed the interspecific character of the F1 hybrids in comparison with B. napus and B. rapa parental lines. All interspecific F1 hybrids had intermediate morphological characteristics typical of both parental components in the generative and vegetative stages. The F1 hybrids of CMS B. napus × B. rapa were sterile, as were those of fertile B. napus × B. rapa. Good seed productivity from CMS B. napus × B. rapa and consecutive back-cross 1 (BC1)–BC3 generations with Chinese cabbage as the recurrent parent was obtained. Plants of the BC1 generation of CMS B. napus × B. rapa were sterile and had highly diversified morphological characteristics, both in the vegetative and generative phases. The BC3 generation of CMS B. rapa was also sterile and more uniform than BC1, with good vigour and most of the typical Chinese cabbage agronomic traits. As a result, a collection of Chinese cabbage genotypes having the CMS trait, which is valuable for the breeding of commercial F1 hybrids, was obtained for the first time in Poland.

Genetic distance of inbred lines of Chinese cabbage and its relationship to heterosis

Chinese cabbage (Brassica rapa L. var. pekinensis) is an important vegetable in Asia. Most Japanese commercial cultivars of Chinese cabbage use an F1 hybrid seed production system because of the high yielding cultivars produced. An efficient method for predicting hybrid performance in the parental generations is desired, and genetic distance between parental lines might be a good indicator of the level of hybrid vigor in a cross. Information concerning the genetic relationships among parental candidate inbred lines is useful for variety protection. The number of DNA markers available that can be used to assess the purity of inbred lines is limited in B. rapa. The aim of this study is to use DNA markers to assess the genetic distance between inbred lines to examine early developmental and yield heterosis so as to develop methods for selecting the best parental lines for the production of hybrids. We screened highly polymorphic SSR and CAPS markers to assess the genetic uniformity of inbred lines and characterize their genetic relationship. We examined the early size and yield heterosis in 32 F1 hybrids of Chinese cabbage. There was a moderate correlation in mid-parent heterosis between leaf size at 21days after sowing and harvested biomass but not in best-parent heterosis. In contrast there was no correlation between genetic distance and mid-parent or best-parent heterosis, indicating that genetic distance does not predict the heterosis phenotype.

Global Gene-Expression Analysis to Identify Differentially Expressed Genes Critical for the Heat Stress Response in Brassica rapa

Genome-wide dissection of the heat stress response (HSR) is necessary to overcome problems in crop production caused by global warming. To identify HSR genes, we profiled gene expression in two Chinese cabbage inbred lines with different thermotolerances, Chiifu and Kenshin. Many genes exhibited >2-fold changes in expression upon exposure to 0.5– 4 h at 45°C (high temperature, HT): 5.2% (2,142 genes) in Chiifu and 3.7% (1,535 genes) in Kenshin. The most enriched GO (Gene Ontology) items included ‘response to heat’, ‘response to reactive oxygen species (ROS)’, ‘response to temperature stimulus’, ‘response to abiotic stimulus’, and ‘MAPKKK cascade’. In both lines, the genes most highly induced by HT encoded small heat shock proteins (Hsps) and heat shock factor (Hsf)-like proteins such as HsfB2A (Bra029292), whereas high-molecular weight Hsps were constitutively expressed. Other upstream HSR components were also up-regulated: ROS-scavenging genes like glutathione peroxidase 2 (BrGPX2, Bra022853), protein kinases, and phosphatases. Among heat stress (HS) marker genes in Arabidopsis, only exportin 1A (XPO1A) (Bra008580, Bra006382) can be applied to B. rapa for basal thermotolerance (BT) and short-term acquired thermotolerance (SAT) gene. CYP707A3 (Bra025083, Bra021965), which is involved in the dehydration response in Arabidopsis, was associated with membrane leakage in both lines following HS. Although many transcription factors (TF) genes, including DREB2A (Bra005852), were involved in HS tolerance in both lines, Bra024224 (MYB41) and Bra021735 (a bZIP/AIR1 [Anthocyanin-Impaired-Response-1]) were specific to Kenshin. Several candidate TFs involved in thermotolerance were confirmed as HSR genes by real-time PCR, and these assignments were further supported by promoter analysis. Although some of our findings are similar to those obtained using other plant species, clear differences in Brassica rapa reveal a distinct HSR in this species. Our data could also provide a springboard for developing molecular markers of HS and for engineering HS tolerant B. rapa.

Resistance to Plasmodiophora brassicae in Brassica rapa and Brassica juncea genotypes From China.

Clubroot disease, caused by Plasmodiophora brassicae Woronin, has become a major problem in cruciferous crops worldwide. Chinese cabbage (Brassica rapa), pak choi (B. rapa), and mustard (B. juncea) are important vegetable crops in China. Development of clubroot-resistant cultivars of these crops is urgently needed. In this study, 71 B. rapa and B. juncea genotypes from China, including cultivars and inbred lines, were evaluated for resistance to three P. brassicae pathotypes. A significant interaction was observed between the P. brassicae pathotypes and the Brassica genotypes. Pathotype 3, as defined on the differentials of Williams, exhibited the weakest virulence on all plant material. By contrast, pathotypes 5 and 6 were both highly pathogenic on most of the tested genotypes. In all, 10 of the 14 Chinese cabbage cultivars were resistant to all three pathotypes, while 4 were resistant only to a specific pathotype. Seven of eight progenies obtained from the selfing of Chinese cabbage cultivars were resistant to pathotype 3 but most were susceptible to pathotypes 5 and 6. Most inbred lines of Chinese cabbage and all inbred lines of pak choi and mustard were susceptible to all three pathotypes but their susceptibility was lower to pathotype 3 than to pathotypes 5 and 6.

Characterization and Gene Co-expression Network Analysis of a Salt Tolerance-related Gene, BrSSR, in Brassica rapa

다양한 비생물적 스트레스 중 토양 염 집적은 식물의 광합성 효율, 생장 및 수확량의 감소를 초래한다. 최근 염 저항성 향상을 위한 많은 유전자들이 보고되고 있다. 본 연구의 목적은 형질전환 배추를 이용하여 아직 기능이 밝혀져 있지 않지만 완전장이 보고된 Brassica rapa Salt Stress Resistance(BrSSR) 유전자의 기능을 검정하는 것이다. BrSSR의 생리적 역할을 분석하기 위해, BrSSR의 과발현 vector인 pSL94 vector를 이용하여 내혼계 배추('CT001')를 형질전환하였다. Quantitative real-time RT-PCR 분석에서 형질전환체의 BrSSR 발현량은 대조군 대비 2.59배까지 증가하였다. 한편, 염 처리 후 표현형 분석에서 BrSSR이 과발현된 형질전환체들이 정상적인 생장을 보여줌으로써 염 스트레스에 내성을 가지는 것을 확인할 수 있었다. Microarray 분석을 통해 구축된 염 스트레스 저항성 관련 유전자들의 발현 네트워크 상에서 BrSSR은 기존에 염 저항성 관련 유전자로 보고되어 있는 ERD15(AT2G41430), protein containing PAM2(AT4G14270), GABA-T(AT3G22200)와 매우 밀접하게 연결되어 있는 것으로 분석되었다. 위 결과들을 바탕으로 BrSSR은 염 스트레스 발생 시 식물의 생장 및 저항성에 관련된 중요한 역할을 하는 것으로 판단된다. Among various abiotic stress factors, soil salinity decreases the photosynthetic rate, growth, and yield of plants. Recently, many genes have been reported to enhance salt tolerance. The objective of this study was to characterize the Brassica rapa Salt Stress Resistance (BrSSR) gene, of which the function was unclear, although the full-length sequence was known. To characterize the role of BrSSR, a B. rapa Chinese cabbage inbred line ('CT001') was transformed with pSL94 vector containing the full length BrSSR cDNA. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that the expression of BrSSR in the transgenic line was 2.59-fold higher than that in the wild type. Analysis of phenotypic characteristics showed that plants overexpressing BrSSR were resistant to salinity stress and showed normal growth. Microarray analysis of BrSSR over-expressing plants confirmed that BrSSR was strongly associated with ERD15 (AT2G41430), a gene encoding a protein containing a PAM2 motif (AT4G14270), and GABA-T (AT3G22200), all of which have been associated with salt tolerance, in the co-expression network of genes related to salt stress. The results of this study indicate that BrSSR plays an important role in plant growth and tolerance to salinity.

Genetic detection of clubroot resistance loci in a new population of Brassica rapa

Clubroot is one of the most serious diseases affecting Brassica crop production worldwide. In order to identify the location of clubroot resistance genes in Chinese cabbage, we constructed a linkage map for an F2 population derived from a cross between a resistant turnip inbred line, 'Siloga' (B. rapa ssp. rapifera), and a susceptible Chinese cabbage inbred line, 'BJN3' (B. rapa ssp. pekinensis). The newly developed genetic map included 207 markers and covered 1151.7 cM. In combination with clubroot resistance tests from the field, the data allowed us to identify three quantitative trait loci of clubroot resistance, using a composite interval mapping method. Clubroot resistance genes QS_B1.1, QS_B3.1, and QS_B8.1 were located on chromosomes A1, A3, and A8, respectively, of B. rapa. Their contribution rates were 8.18%, 70.55%, and 7.28%, respectively. QS_B1.1 was a novel locus of clubroot resistance, independent of any published clubroot-resistance loci. QS_B3.1 was first detected in 'Siloga,' mapped to the previous CRb and CRa region, whereas QS_B8.1 was closely linked to Crr1b. Epistatic interactions with additive effects were detected between QS_B3.1 and QS_B8.1.

Mapping the BrPur gene for purple leaf color on linkage group A03 of Brassica rapa

The purple-leaf phenotype in Chinese cabbage is due to the accumulation of anthocyanin. To investigate the pattern of inheritance of this trait in Brassica rapa, F1, F2 and backcross (BC) populations were constructed by crossing 09N-742, a pak-choi inbred line that has purple leaves, with a green-leaved Chinese cabbage inbred line, 09-680. Using a segregating F2 population, we identified a single dominant gene, BrPur, for purple leaf, and mapped the gene to a locus on linkage group A03 of B. rapa. Furthermore, sequences from BAC clones and other sources were used to develop molecular marker loci that are tightly linked to BrPur by using a BC1 population of 1,152 individuals. BrPur was assigned to a locus between Indel markers BVRCPI613 and BVRCPI431, which defined a genetic interval of 0.6 cM and a genomic region of 54.87 kb. Sequence analysis of this chromosomal region revealed seven open reading frames. These results provide a foundation for map-based cloning, identification, and functional analysis of the BrPur gene in B. rapa.

Identification of Novel QTLs for Isolate-Specific Partial Resistance to Plasmodiophora brassicae in Brassica rapa

Plasmodiophora brassicae, the causal agent of clubroot disease of the Brassica crops, is widespread in the world. Quantitative trait loci (QTLs) for partial resistance to 4 different isolates of P. brassicae (Pb2, Pb4, Pb7, and Pb10) were investigated using a BC1F1 population from a cross between two subspecies of Brassica rapa, i.e. Chinese cabbage inbred line C59-1 as a susceptible recurrent parent and turnip inbred line ECD04 as a resistant donor parent. The BC1F2 families were assessed for resistance under controlled conditions. A linkage map constructed with simple sequence repeats (SSR), unigene-derived microsatellite (UGMS) markers, and specific markers linked to published clubroot resistance (CR) genes of B. rapa was used to perform QTL mapping. A total of 6 QTLs residing in 5 CR QTL regions of the B. rapa chromosomes A01, A03, and A08 were identified to account for 12.2 to 35.2% of the phenotypic variance. Two QTL regions were found to be novel except for 3 QTLs in the respective regions of previously identified Crr1, Crr2, and Crr3. QTL mapping results indicated that 1 QTL region was common for partial resistance to the 2 isolates of Pb2 and Pb7, whereas the others were specific for each isolate. Additionally, synteny analysis between B. rapa and Arabidopsis thaliana revealed that all CR QTL regions were aligned to a single conserved crucifer blocks (U, F, and R) on 3 Arabidopsis chromosomes where 2 CR QTLs were detected in A. thaliana. These results suggest that some common ancestral genomic regions were involved in the evolution of CR genes in B. rapa.