Black rot, a disease of significance affecting vegetable Brassica crops, is primarily caused by the bacterium Xanthomonas campestris pv. campestris (Xcc). When the disease spreads extensively in the field, it can lead to substantial yield losses, particularly under favorable environmental conditions. Controlling the spread of this disease is challenging, and the primary approach involves utilizing resistant cultivars or disease-free seeds. Among the various methods available for identifying different Xcc races, Polymerase Chain Reaction (PCR)-based molecular markers have proven to be highly reliable. To date, the PCR method has successfully identified Xcc races 1 to 7. In this study, molecular markers were developed for races 4 and 9 through the sequencing and alignment of the whole genome sequences of Xcc races, closely related Xanthomonas campestris (Xc) pathovars, and two Xanthomonas species. These designed markers were subsequently validated by PCR with bacterial genomic DNA samples from Xcc races and 7 other bacteria. The results indicated successful amplification only for race 4 and race 9, yielding amplicon sizes of 1080 bp and 830 bp, respectively, while the other strains failed to amplify. Furthermore, the amplicons from races 4 and 9 were cloned and sequenced, confirming that both races exhibited matching sequences after alignment. Consequently, the molecular marker method offers a rapid and efficient means of differentiating between Xcc races 4 and 9 within a few hours, presenting itself as a viable alternative to conventional methods that rely on the use of differential cultivars of Brassicaceae for identifying Xcc races.

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Black rot of Brassica crops is the most devastating disease which causes substantial yield reduction of cabbage throughout the world. The use of resistant cabbage cultivars could be inexpensive and effective measure to combat this destructive disease. We screened cabbage inbred lines for black rot disease resistance through bioassay and identified some novel lines that showed race-specific resistance to Xanthomonas campestris pv. campestris (Xcc) races. The pathogenicity test revealed that out of 27 cabbage lines, one (SCNU-C-4074), six (SCNU-C-3631, SCNU-C-3637, SCNU-C-3639, SCNU-C-4072, SCNU-C-4073 and SCNU-C-3273), two (SCNU-C-3273 and SCNU-C-4118), two (SCNU-C-3270 and SCNU-C-4118), two (SCNU-C-3470 and SCNU-C-41148) and four (SCNU-C-107, SCNU-C-3270, SCNU-C-3470 and SCNU-C-4059) were shown to be resistant to Xcc races 1, 2, 3, 5, 6 and 7, respectively while none of these showed resistance against race 4. Furthermore, these resistant and susceptible lines were evaluated by previously reported molecular markers for black rot resistance. The molecular screening results were also revealed the existence of race-specific resistance in these cabbage lines. This result will help Brassica breeder to develop race-specific black rot resistant cabbage cultivars.

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Through high throughput shotgun proteomics approach, the proteome of seedling leaf of Brassica rapa L. was identified. From three biological replications, a total of 2,122 non-redundant proteins of Brassica rapa L seedling leaf were identified, with a wide range and unbiased physiochemical properties. Their pI values ranged from pH 4.27 (Bra004590) to pH 11.81 (Bra013905). Their molecular weight (MW) ranged from 5.6 kDa (Bra006908) to 534.5 kDa (Bra028068). Gene ontology enrichment analysis revealed that these proteins were associated with cellular process, metabolic process, and enriched catalytic activity compared to whole brassica proteins. The highest presented protein in Brassica rapa seedling leaf was RuBisCO, accounting for 11.56% of total leaf proteins. Also, many ribosomal proteins were identified. The relative amount of all ribosomal proteins comprised 8.47% of total leaf proteins. The relative amount of two RuBisCO and ribosomal proteins was about 20% of total leaf proteins. Thus to detect proteins presenting low abundance, additional fractionating procedure to remove RuBisCO and ribosomal proteins is required.

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The purpose of this study was to produce doubled haploid red color heading type Chinese cabbage using isolated microspore culture. Genotypic differences in embryogenic response and regenerative ability of microspore-derived embryos to plants were observed. A high rate of plant regeneration from microspore-derived embryos was achieved by an improved protocol involving replacement of culture media and adjustment of heat shock temperature. More than 60% of regenerated plants were spontaneous doubled haploids. Haploids were characterized by short and malformed stamen and few viable pollen grains as compared to spontaneous doubled haploids and dihaploids. The seeds harvested from the 142 spontaneous doubled haploid plants were designated as H1; 25 seeds in each population was grown and characterized. The H1 generation has been shown to display considerable phenotypic variation and high purity. These spontaneous doubled haploids may be directly exploited as new varieties in a red colored Chinese cabbage breeding program.

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Cytoplasmic male sterility (CMS) is a phenomenon specific to plant reproduction that has been extensively exploited for hybrid seed production. Orf456 was previously identified as a candidate gene mediating male sterility in pepper. To gain further insight of this candidate gene, we carried out experiments to transform Chinese cabbage (Brassica rapa L.). About 30 T₁ transgenic lines were obtained and approximately 50 % of T₁ transgenic Chinese cabbage lines showed male-sterility. To evaluate pollen viability, three different approaches including plasmolysis test, staining pollen and in vitro germination assay were used. Analysis of the CMS transgenic lines showed that trasgenic Chinese cabbages produced aberrant pollen development while some were unable to produce pollen. In conclusion, the mitochondrial orf456 gene could induce partial male sterility in transgenic Chinese cabbage. Nevertheless, how the orf456 gene precisely functions to induce male sterility and its biochemical function remains to be discovered.

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Soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum (Pcc), is one of the most devastating diseases affecting the cruciferous plants industry worldwide. In our previous study, the soft rot-resistant transgenic Chinese cabbage (Brassica rapa L.) plants were produced via constitutively overexpressing a human cathelicidin antimicrobial peptide (hCAP18/LL-37). To unravel the molecular mechanisms underlying Pcc resistance of the transgenic plants, this study compares the global transcriptional profile of untransformed line (WT) and the transgenic lines (TG23, TG34) through hybridization with KBGP-24K, Chinese cabbage GeneChip. In total, 1,415 differentially expressed genes (DEGs) were identified, 910 of which were up-regulated, while 505 were down-regulated. The DEGs were classified into 31 categories after Gene ontology (GO) annotation, in which 68 genes are in response to stimulus and are involved in immune system process, 12 genes are related to cell wall, and 13 genes belong to transcription factors. These genes and those related to toxin and terpenoid metabolism, glutathione metabolism, biosynthesis of phenylpropanoids, and plant hormones were hypothesized to play major roles in the soft rot resistance of transgenic lines (TG23, TG34). Semiquantitative RT-PCR analysis showed that the transcript levels of several candidate genes in TG23 and TG34 were significantly higher than in WT both before and after Pcc inoculation, indicating their potential association with soft rot disease.

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