Plastic film mulching is a common practice to increase vegetable yields. However, the effects of plastic mulch films on different vegetable fields remain largely unknown. In the current study, biodegradable mulch films (BDMs) and polyethylene mulch films (PEMs) were used to explore their effects on three vegetable fields (broccoli, chili pepper and garlic). Compared with bare soil, both BDMs and PEMs could effectively maintain higher exchangeable potassium contents in rhizosphere soil, and significantly increase various soil properties and microbial abundance both in rhizosphere and non-rhizosphere soils. In terms of yields, the treatments of BDMs and PEMs effectively increased broccoli, chili pepper and garlic yields by 17.6–19.0%, 21.3–23.3% and 26.0–29.5%, respectively, when compared with the bare soil. Our initial characterization provided insights into the effects of BMDs and PEMs on soil properties, yields and diversity of rhizosphere and non-rhizosphere microbial communities in three vegetable fields.

Basic helix–loop–helix (bHLH) transcription factor (TF) family is commonly found in eukaryotes, which is one of the largest families of regulator proteins. It plays an important role in plant growth and development, as well as various biotic and abiotic stresses. However, a comprehensive analysis of the bHLH family has not been reported in Brassica oleracea. In this study, we systematically describe the BobHLHs in the phylogenetic relationships, expression patterns in different organs/tissues, and in response to chilling stress, and gene and protein characteristics. A total of 234 BobHLH genes were identified in the B. oleracea genome and were further clustered into twenty-three subfamilies based on the phylogenetic analyses. A large number of BobHLH genes were unevenly located on nine chromosomes of B. oleracea. Analysis of RNA-Seq expression profiles revealed that 21 BobHLH genes exhibited organ/tissue-specific expression. Additionally, the expression of six BobHLHs (BobHLH003, -048, -059, -093, -109, and -148) were significantly down-regulated in chilling-sensitive cabbage (CS-D9) and chilling-tolerant cabbage (CT-923). At 24h chilling stress, BobHLH054 was significantly down-regulated and up-regulated in chilling-treated CS-D9 and CT-923. Conserved motif characterization and exon/intron structural patterns showed that BobHLH genes had similar structures in the same subfamily. This study provides a comprehensive analysis of BobHLH genes and reveals several candidate genes involved in chilling tolerance of B. oleracea, which may be helpful to clarify the roles of bHLH family members and understand the regulatory mechanisms of BobHLH genes in response to the chilling stress of cabbage.

Squamosa promoter-binding protein-like (SPL) is a major family of plant-specific transcription factor, which is involved in multiple biological processes, such as plant growth and development, hormone response, light response and stress response. Therefore, it has been profoundly significant to systematically analyze the SPL Transcription Factors family in Brassica oleracea. In this study, a total of 33 BoSPLs were identified in the B. oleracea genome, and they were further divided into six subgroups based on the phylogenetic tree constructed from the SPL proteins of B. oleracea, B. rapa and Arabidopsis thaliana. The expression profile of BoSPLs in different organs/tissues showed that a large number of BoSPLs were expressed in the callus, root, stem, leaf, bud, flower and silique. In addition, the expression levels of two BoSPLs (BoSPL9b and BoSPL10b) were up-regulated in chilling tolerance cabbage ‘CT-923’ at 6 h after chilling stress when compared with normal treatment (mock), while two BoSPLs (BoSPL9b and BoSPL15a) in chilling sensitive cabbage ‘CS-D9’, five BoSPLs (BoSPL1, -9a, -9b, -10b, -11b) in ‘CT-923’ and two BoSPLs (BoSPL9b and BoSPL16a) in ‘CS-D9’ were up-regulated after 24 h chilling stress, indicated that these genes may play an important role in the chilling-tolerance of cabbage. We analyzed the characteristics of BoSPLs and provided the basis for further functional research.

Yellow leaf mutations have been widely used to study the chloroplast structures, the pigment synthesis, the photosynthesis mechanisms and the chlorophyll biosynthesis pathways across various species. For this study, a spontaneous mutant with the yellow leaf color named 96-140YBM was employed to explore the primary genetic elements that lead to the variations in the leaf color of hot peppers. To identify the pathways and genes associated with yellow leaf phenotypes, we applied sequencing-based Bulked Segregant Analysis (BSA-Seq) combined with BSR-Seq. We identified 4167 differentially expressed genes (DEGs) in the mutant pool compared with the wild-type pool. The results indicated that DEGs were involved in zeatin biosynthesis, plant hormone signal transduction, signal transduction mechanisms, post-translational modification and protein turnover. A total of 437 candidates were identified by the BSA-Seq, while the BSR-Seq pinpointed four candidate regions in chromosomes 8 and 9, containing 222 candidate genes. Additionally, the combination of BSA-Seq and BSR-Seq showed that there were 113 overlapping candidate genes between the two methods, among which 8 common candidates have been previously reported to be related to the development of chloroplasts, the photomorphogenesis and chlorophyll formation of plant chloroplasts and chlorophyll biogenesis. qRT-PCR analysis of the 8 common candidates showed higher expression levels in the mutant pool compared with the wild-type pool. Among the overlapping candidates, the DEG analysis showed that the CaKAS2 and CaMPH2 genes were down-regulated in the mutant pool compared to the wild type, suggesting that these genes may be key contributors to the yellow leaf phenotype of 96-140YBM. This research will deepen our understanding of the genetic basis of leaf color formation and provide valuable information for the breeding of hot peppers with diverse leaf colors.

Cabbage (Brassica oleracea var. capitata) is a biennial plant. Gene editing technology has not been extensively studied in this species. In this study, we report the induction of highly efficient and heritable transgenic roots in cabbage using the CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9) gene editing system, followed by the regeneration of whole plants from these transgenic roots. We designed and constructed a CRISPR/Cas9 gene editing vector targeting the immune regulatory gene PUB13 (PLANT U-BOX 13) and introduced it into plants through Agrobacterium-mediated transformation. By performing targeted mutations on PUB13, transgenic roots were obtained, the optimal TDZ (Thidiazuron) concentration for bud induction (0.9 mg·L⁻¹) was determined, and then an efficient transformation protocol from transgenic roots to plants was established, leading to the regeneration of gene-edited plants. In summary, we successfully generated gene-edited cabbage (Brassica oleracea var. capitata) plants through PUB13 gene mutagenesis using an innovative transgenic root regeneration approach. A new pathway for obtaining gene-edited cabbage plants was established.