A tissue-specific and developmentally expressed gene was isolated from Chinese cabbage (Brassica rapa L. ssp. pekinensis), designated BrREF (B. rapa Rubber elongation factor). BrREF transcripts were expressed at high levels in seedlings and at low levels in flower buds and roots. To study the activity of this promoter, the 2.2 kb upstream sequence of BrREF gene was fused to a β-glucuronidase (GUS) reporter gene and was introduced into Arabidopsis thaliana and B. napus by Agrobacterium-mediated transformation. Strong expression of GUS driven by the BrREF promoter was detected in the cotyledons and hypocotyls of transgenic plant seedlings, but GUS expression was weak in roots, excluding the root tips. GUS expression in the cotyledons and hypocotyls decreased dramatically as the seedlings matured and was not detected in the tissues of mature plants. During floral development, GUS expression was observed in immature anthers. These findings suggest that the BrREF promoter can modulate the tissue-specific and developmental expression of gene at the early stages of growth and development.

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• The Characterization of Constitutive Promoters in Chrysanthemum (Chrysanthemum morifolium Ramat)
  Eun Jung Suh, So Youn Won, Seong-Kon Lee, Sang Ryeol Park
  Korean Journal of Breeding Science.2024; 56(3): 179.     CrossRef

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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• Microspore Embryogenesis Efficiency in Chinese Cabbage: The Predominant Role of Genotypic Variation
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• Possibilities of biotechnological methods in breeding of vegetable crops at the VIR Laboratory of Breeding and Cell Technologies
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• Influencing factors and physiochemical changes of embryogenesis through in vitro isolated microspore culture in Brassica species
  Yan-Qi Dong, Yu-Hong Gao, Te Zhao, Guang-Qian Ren, Yan-Li Liu, Bin Guan, Rong-Xian Jin, Fei Gao, Yan-Li Zhang, Xiu-Fang Tan, Hong-Cai Zhu, Yu-Hong Zhang, Jin-Xia Zhang, Dong Peng, Yu-Xin Yan
  Biologia.2021; 76(9): 2629.     CrossRef
• Non-ionic surfactants improved microspore embryogenesis and plant regeneration of recalcitrant purple flowering stalk (Brassica campestris ssp. chinensis var. purpurea Bailey)
  Yiming Gao, Junxiang Jia, Jialin Cong, Yuying Ma, Hui Feng, Yun Zhang
  In Vitro Cellular & Developmental Biology - Plant.2020; 56(2): 207.     CrossRef
• Microspore embryogenesis in Brassica: calcium signaling, epigenetic modification, and programmed cell death
  Behzad Ahmadi, Medya Ahmadi, Jaime A. Teixeira da Silva
  Planta.2018; 248(6): 1339.     CrossRef

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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• Variations in chlorosis and potential usefulness of alloplasmic Brassica rapa with the cytoplasm of male sterile Brassica juncea
  Caitao Chang, Deling Sun, Kana Hondo, Fumika Kakihara, A.‐M. Chevre
  Plant Breeding.2014; 133(5): 620.     CrossRef

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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• Enhanced resistance to bacterial pathogen in transgenic tomato plants expressing cathelicidin antimicrobial peptide
  Yu-Jin Jung
  Biotechnology and Bioprocess Engineering.2013; 18(3): 615.     CrossRef