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The quality of green leafy vegetables is easily lost during the postharvest period. The effect of exogenous 24‐epibrassinolide (EBR) pretreatment on the quality of wucai was evaluated in the present study. Wucai plants were sprayed twice with 0.1 μM EBR before harvesting. Two storage temperatures were tested: 25°C and 4°C. At 4°C, EBR pretreatment significantly delayed the degradation of the pigment and plant water loss. Furthermore, we measured the activity of key enzymes of the ascorbic acid (AsA)‐glutathione (GSH) cycle, the content of the main metabolites, and the expression of the AsA metabolism‐related genes in leaves. The results indicated that all three plants showed stronger antioxidant capacity after EBR pretreatment. At 4°C and 25°C, the storage time of wucai was 20 days and 7 days after EBR treatment, while the samples could be stored for 14 days and 4 days without EBR treatment application, respectively. At 4°C, the nutritional properties of wucai pretreated with EBR, such as total free amino acids, total soluble sugar, and cellulose contents, were higher than those of the control, while the content of nitrite and lignin was lower than that of the control. Hence, EBR pretreatment was able to enhance the antioxidant capacity of wucai, maintain normal leaf color and shape during storage, and delay the decline of nutritional properties; therefore, EBR pretreatment has potential commercial value for prolonging the market life of wucai., EBR pretreatment was able to enhance the antioxidant capacity of wucai, maintain normal leaf color and shape during storage, and delay the decline of nutritional properties; therefore, EBR pretreatment has potential commercial value for prolonging the market life of wucai.

Zizania latifolia is a perennial aquatic vegetable, whose symbiosis with the fungus Ustilago esculenta (member of Basidiomycota, class Ustilaginaceae) results in the establishment of swollen gall formations. Here, we analyzed symbiotic relations of Z. latifolia and U. esculenta, using a triadimefon (TDF) treatment and transcriptome sequencing (RNA-seq). Specifically, accurately identify the whole growth cycle of Z. latifolia. Microstructure observations showed that the presence of U. esculenta could be clearly observed after gall formation but was absent after the TDF treatment. A total of 17,541 differentially expressed genes (DEGs) were identified, based on the transcriptome. According to gene ontology term and Kyoto Encyclopedia of Genes and Genomes pathway results, plant hormone signal transduction, and cell wall–loosening factors were all significantly enriched due to U. esculenta infecting Z. latifolia; relative expression levels of hormone-related genes were identified, of which downregulation of indole 3-acetic acid (IAA)-related DEGs was most pronounced in JB_D versus JB_B. The ultra–high performance liquid chromatography analysis revealed that IAA, zeatin+trans zeatin riboside, and gibberellin 3 were increased under U. esculenta infection. Based on our results, we proposed a hormone–cell wall loosening model to study the symbiotic mechanism of gall formation after U. esculenta infects Z. latifolia. Our study thus provides a new perspective for studying the physiological and molecular mechanisms of U. esculenta infection of Z. latifolia causing swollen gall formations as well as a theoretical basis for enhancing future yields of cultivated Z. latifolia. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law. 2021

Chilling stress during the growing season could cause a series of changes in wucai (Brassica campestris L.). WS-1 (chilling-tolerant genotype) and Ta2 (chilling-sensitive genotype) were sampled in present study to explore the chilling tolerance mechanisms. Our results indicated that photosynthetic parameters exhibited lower level in Ta2 than in WS-1 under chilling stress. The rapid chlorophyll fluorescence dynamics curve showed that chilling resulted in a greater inactivation of photosystem II reaction center in Ta2. Reactive oxygen species and malondialdehyde content of chloroplast in Ta2 were higher than WS-1. The ascorbate–glutathione cycle in chloroplast of WS-1 played a more crucial role than Ta2, which was confirmed by higher activities of antioxidant enzymes including Ascorbate peroxidase, Glutathione reductase, Monodehydroascorbate reductase and Dehydroascorbate reductase and higher content of AsA and GSH. In addition, the ultrastructure of chloroplasts in Ta2 was more severely damaged. After low temperature stress, the shape of starch granules in Ta2 changed from elliptical to round and the volume became larger than that of WS-1. The thylakoid structure of Ta2 also became dispersed from the original tight arrangement. Combined with our previous study under heat stress, WS-1 can tolerant both chilling stress and heat stress, which was partly due to a stable photosynthetic system and the higher active antioxidant system in plants, in comparison to Ta2.

The effects of plant hormones on plant growth of Zizania latifolia are still considered to be the difficult and hot spots. The present study was carried out to investigate the regulation mechanism of exogenous indole-3-acetic acid (IAA) and IAA transport inhibitor (2,3,5-triiodobenzoic acid, TIBA) on yield, photosynthesis capacity, carbon assimilation and interaction of plant-fugal of Ustilago esculenta Henn. These results displayed that exogenous IAA led to significant increases in gall growth rate, swollen gall volume and biomass accumulation, promoting yield per unit of swollen gall in Z. latifolia. Exogenous IAA could improve the net photosynthetic rate (Pn), which contributed to higher stomatal conductance (Gs) and transpiration rate (Tr) in plant. TIBA treatment markedly inhibited the plant growth and photosynthetic rate. Contents of sucrose, glucose, fructose and total soluble sugar by IAA were significantly higher than control, whereas these parameters were declined by TIBA. Exogenous TIBA resulted in accumulation of starch (unfavorable to sugar metabolism) and crude fiber contents (declined swollen gall quality). Additionally, it was firstly reported the stimulation of exogenously applied of IAA and TIBA on plant growth and hyphae growth of U. esculenta. Microscopic analyses revealed that abundant hyphae were well-distributed closely to cell wall after IAA treatment, which could produce endogenous plant hormones, promoting the swollen cell structure enlargement. And hyphal aggregation over-growth by TIBA treatment indicated the vigorously increase of hyphae was surpassed the plant self-growth. These changes were greatly contributed to the increase of endogenous IAA content. The utilization of exogenous IAA and TIBA in present study proved IAA played a dominant role in regulating Z. latifolia growth.

We developed a novel Agrobacterium-mediated anther transformation for Wucai in planta, and in this procedure, the male germ line was the predominant target. Wucai (Brassica campestris L.), a variant of non-heading Chinese cabbage, is widely cultured in China and only improved by classic breeding methods. Here, a novel and efficient in planta Agrobacterium-mediated anther transformation method is developed based on the optimization of several factors that affect anther transformation. After optimization, transformation with the manual pollination application led to increased transient GUS expression in anthers (reaching 91.59%) and the transformation efficacies in planta (0.59–1.56% for four commercial cultivars). The stable integration and inheritance of the transgenes were further examined by molecular and genetic analyses. Three T2 transgenic lines presented a segregation ratio of 3:1, which was consistent with the Mendelian feature of a single dominant gene. In addition, the GUS histochemical assay and genetic crossing analysis revealed that the male germ line was the predominant target in this transformation. This optimized transformation system could provide a useful tool for both the improvement of cultivar qualities and investigation of functional genes in Wucai.

Brassica rapa is an important Chinese vegetable crop that is beneficial to human health. The primary factor affecting B. rapa yield is low temperature, which promotes bolting and flowering, thereby lowering its commercial value. However, quickened bolting and flowering can be used for rapid breeding. Therefore, studying the underlying molecular mechanism of vernalization in B. rapa is crucial for solving production-related problems. Here, the transcriptome of two B. rapa accessions were comprehensively analyzed during different vernalization periods. During vernalization, a total of 974,584,022 clean reads and 291.28 Gb of clean data were obtained. Compared to the reference genome of B. rapa, 44,799 known genes and 2280 new genes were identified. A self-organizing feature map analysis of 21,035 differentially expressed genes was screened in two B. rapa accessions, &lsquo, Jin Wawa&rsquo, and &lsquo, Xiao Baojian&rsquo, The analysis indicated that transcripts related to the plant hormone signal transduction, starch and sucrose metabolism, photoperiod and circadian clock, and vernalization pathways changed notably at different vernalization periods. Moreover, different expression patterns of TPS, UGP, CDF, VIN1, and seven hormone pathway genes were observed during vernalization between the two accessions. The transcriptome results of this study provide a new perspective on the changes that occur during B. rapavernalization, as well as serve as an excellent reference for B. rapa breeding.

Cucumber is a major vegetable crop grown in facilities, and its seedlings are sensitive to salinity. Thus, it is important for cucumber cultivation to research the effects of Ca(NO3)2. In this study, we investigated salt-sensitive Chunqiu (CQ) and salt-tolerant BoMei 49 (BM) seedlings, the changes to the enzymes involved in the mitochondria antioxidant system in the seedlings, and the changes in the MPTP (Mitochondrial Permeability Transition Pore) opening, mitochondrial membrane fluidity, mitochondrial membrane potential depolarization, mitochondrial electron transfer chain, and NO metabolism in response to Ca(NO3)2. Ca(NO3)2 stresses inhibited the growth of the cucumber seedlings, which showed a reduced root length, a decreased surface area and a decreased number of root tips, but a significantly increased root diameter. The mitochondrial malondialdehyde (MDA) content, H2O2 content, and O2&minus, producing rate of the two cucumber roots were gradually increased in the Ca(NO3)2 treatment. The activity of superoxide dismutase (SOD) and peroxidation enzyme (POD) gradually increased, and catalase (CAT) activity gradually decreased. The electron transport chain activity of &ldquo, BM&rdquo, was higher than &ldquo, CQ&rdquo, under Ca(NO3)2 stress. Ca(NO3)2 stress destroyed the membrane structure of the mitochondria, so that the mitochondrial body tended to bend, causing double-membrane digestion and a hollow interior. Under the stress of Ca(NO3)2, the NO3- content of the seedlings increased significantly. The contents of NH4+ and NO, as well as the activities of nitrate reductase (NR), nitrite reductase (NIR), and nitric oxide synthase (NOS) decreased significantly. These findings provide physiological insights into root damage in response to salt stress for salt tolerance in cucumber.

High temperature (HT) is a major environmental stress limiting oversummer production of nonheading Chinese cabbage (NHCC, Brassica campestris ssp. chinensis Makino). In the present study, the effects of HT on photosynthetic capacity, including light reaction and carbon assimilation, were completely investigated in two NHCC, ‘xd’ (heat-tolerant), and ‘sym’ (heat-susceptible). The two genotypes showed significant differences in plant morphology, photosynthetic capacity, and photosynthate metabolism (carboassimilation). HT caused a decrease in photosynthesis, chlorophyll contents, and photochemical activity in NHCC. However, these main photosynthetic-related parameters, including net photosynthetic rate (PN), maximal photochemical efficiency of PSII (Fv/Fm), and total chlorophyll content in ‘xd’, were significantly higher than those of ‘sym’ plants. The antioxidant contents and antioxidative enzyme activities of ascorbic acid-reduced glutathione cycle in the chloroplast of ‘xd’ were significantly higher than those of ‘sym’. Microscopic analyses revealed that HT affected the structure of photosynthetic apparatus and membrane integrity to a different extent, whereas ‘xd’ could maintain a better integrated chloroplast shape and thylakoid. Inhibited light reaction also hampered carbon assimilation, resulting in a decline of carboxylation efficiency and imbalance of carbohydrate metabolism. However, larger declined extents in these data were presented in ‘sym’ (heat-susceptible) than ‘xd’ (heat-tolerant). The heat-tolerant genotype ‘xd’ had a better capacity for self-protection by improved light reaction and carbon assimilation responding to HT stress.

Light is one of the most important abiotic factors for most plants, which affects almost all growth and development stages. In this study, physiological indicators suggest that the application of exogenous Ca2+ improves photosynthesis and changes phytohormone levels. Under weak light, photosynthetic parameters of the net photosynthetic rate (PN), stomatal conductance (Gs), and transpiration rate (Tr) decreased, the antioxidation systems peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) reduced, the degrees of malondialdehyde (MDA), H2O2, and superoxide anion (O2&minus, ) free radical damage increased, while exogenous Ca2+ treatment was significantly improved. RNA-seq analysis indicated that a total of 13,640 differently expressed genes (DEGs) were identified and 97 key DEGs related to hormone, photosynthesis, and calcium regulation were differently transcribed. Gene ontology (GO) terms and Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses, plant hormone signal transduction, photosynthesis, carbon metabolism, and phenylpropanoid biosynthesis were significantly enriched. Additionally, quantitative real-time PCR (qRT-PCR) analysis confirmed some of the key gene functions in response to Ca2+. Overall, these results provide novel insights into the complexity of Ca2+ to relieve injuries under weak light, and they are helpful for potato cultivation under weak light stress.

The genotype WS-1, previously identified from novel wucai germplasm, is tolerant to both low-temperature (LT) and high-temperature (HT) stress. However, it is unclear which signal transduction pathway or acclimation mechanisms are involved in the temperature-stress response. In this study, we used the proteomic method of tandem mass tag (TMT) coupled with liquid chromatography-mass spectrometry (LC-MS/MS) to identify 1022 differentially expressed proteins (DEPs) common to WS-1, treated with either LT or HT. Among these 1022 DEPs, 172 were upregulated in response to both LT and HT, 324 were downregulated in response to both LT and HT, and 526 were upregulated in response to one temperature stress and downregulated in response to the other. To illustrate the common regulatory pathway in WS-1, 172 upregulated DEPs were further analyzed. The redox homeostasis, photosynthesis, carbohydrate metabolism, heat-shockprotein, and chaperones and signal transduction pathways were identified to be associated with temperature stress tolerance in wucai. In addition, 35S:BcccrGLU1 overexpressed in Arabidopsis, exhibited higher reduced glutathione (GSH) content and reduced glutathione/oxidized glutathione (GSH/GSSG) ratio and less oxidative damage under temperature stress. This result is consistent with the dynamic regulation of the relevant proteins involved in redox homeostasis. These data demonstrate that maintaining redox homeostasis is an important common regulatory pathway for tolerance to temperature stress in novel wucai germplasm.

People are increasingly concerned about the nutritional value of the foods they consume, and consequently the best cultivation practices are needed to produce crops that contain high functional nutrients [1].

Wucai is a biannual leafy vegetable that is more sensitive to cold than most Brassicaceae vegetables. Here, phenotypic differences in wucai were investigated to identify those genotypes with high cold tolerance at low temperature, which could allow their cultivation in cold climates. In all, on the basis of PIabs, 20 wucai genotypes (10 lower PIabs and 10 higher PIabs) were selected from a collection of 124 genotypes. Both W16-13 and SW-3 (higher PIabs) and the LS-6 and W15-16 (lower PIabs) were screened from the 20 wucai genotypes according to several key plant physiological traits: net photosynthetic rate, electrolyte leakage, chilling injury index, specific leaf area, malondialdehyde contents, and PIabs. To further verify the reliability of PIabs, four genotypes were grown under low temperature gradient conditions and their morphological indexes, chlorophyll content, total antioxidant capacity, fluorescence transients (OJIP transients), leaf cell ultrastructure, and gene expression were measured. These indicators strongly demonstrated that the W16-13 and SW-3, which had higher PIabs, possessed higher resistance to cold stress, while both LS-6 and W15-16 were sensitive to cold. As this study shows, the easily measured performance index, PIabs, could be applied to wucai genotypes to screen for one or more varieties characterized by higher cold tolerance.

High temperature stress (>30 °C) from late summer to early fall is one of the main limiting factors in wucai ( Brassica campestris L.) growth in southern China. To assess the osmotic and antioxidative reponses of two wucai genotypes (WS-6 and WS-1) differing in heat tolerance, three-week-old seedlings were subjected to three heat-stress treatments for 5 days. Heat treatments above at 30 °C resulted in significant decreases in leaf area and leaf relative water content (RWC) in both genotypes, but being more in heat-sensitive WS-6. This was due to greater accumulations of osmolytes including soluble sugar, soluble protein and proline (Pro), as well as higher levels of reduced ascorbic acid (AsA) and glutathione (GSH) in heat-tolerant WS-1 under heat stress as compared to those in heat-sensitive WS-6. It is worthy noting that mild hyperthermia (30 °C) caused significant increases of AsA and GSH in both genotypes, which might be resulted from their enhanced activities of monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR), helping recycle AsA and GSH. In contrast, the AsA and GSH levels of both genotypes were decreased markedly at 40 °C, especially in heat-sensitive WS-6, despite that the enzymes (DHAR and GR) activities involved in their regeneration remained high levels as compared to their respective controls. In addition, larger decrease in total antioxidant capacity (T-AOC) by heat stress was observed in WS-6 than in WS-1. Our results indicated that the heat-sensitive and the heat-tolerant genotype of wucai might possess the same pathway but different capacities of antioxidant defences to cope with heat stress.

Background Wucai (Brassica campestris L. ssp. chinensis var. rosularis Tsen) is a variant of nonheading Chinese cabbage (Brassica campestris L.), which is one of the major vegetables in China. Cytoplasmic male sterility (CMS) has been used for Wucai breeding in recent years. However, the underlying molecular mechanism of Wucai CMS remains unclear. In this study, the phenotypic and cytological features of Wucai CMS were observed by anatomical analysis, and a comparative transcriptome analysis was carried out to identify genes related to male sterility using Illumina RNA sequencing technology (RNA-Seq). Results Microscopic observation demonstrated that tapetum development was abnormal in the CMS line, which failed to produce fertile pollen. Bioinformatics analysis detected 4430 differentially expressed genes (DEGs) between the fertile and sterile flower buds. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to better understand the functions of these DEGs. Among the DEGs, 35 genes (53 DEGS) were implicated in anther and pollen development, and 11 genes were involved in pollen cell wall formation and modification; most of these showed downregulated expression in sterile buds. In addition, several genes related to tapetum development (A6, AMS, MS1, MYB39, and TSM1) and a few genes annotated to flowering (CO, AP3, VIN3, FLC, FT, and AGL) were detected and confirmed by qRT-PCR as being expressed at the meiosis, tetrad, and uninucleate microspore stages, thus implying possible roles in specifying or determining the fate and development of the tapetum, male gametophyte and stamen. Moreover, the top four largest transcription factor families (MYB, bHLH, NAC and WRKY) were analyzed, and most showed reduced expression in sterile buds. These differentially expressed transcription factors might result in abortion of pollen development in Wucai. Conclusion The present comparative transcriptome analysis suggested that many key genes and transcription factors involved in anther development show reduced gene expression patterns in the CMS line, which might contribute to male sterility in Wucai. This study provides valuable information for a better understanding of CMS molecular mechanisms and functional genome studies in Wucai. Electronic supplementary material The online version of this article (10.1186/s12864-018-5331-4) contains supplementary material, which is available to authorized users.

The hot climate of southern China from late summer to early fall is one of the major factors limiting the yield and quality of wucai (Brassica campestris L.). To study the effects of high temperature on the growth and development of wucai, two cultivars (heat-sensitive ‘WS-6’ and heat-tolerant ‘WS-1’) were subjected to three heat stress treatments for 5 days. The two cultivars were evaluated for plant growth, photosynthetic characteristics, oxidative damage, and activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Heat stress treatments higher than 30 °C resulted in significant changes for both cultivars in plant growth (plant height, stem diameter, and single plant weight) and photosynthetic performance [net photosynthetic rate (P N), stomatal conductance (g s), intercellular CO2 concentration (C i), and leaf transpiration rate (E)]. The chlorophyll (Chl) content was markedly reduced by heat stress, inhibiting photochemical activity. However, larger decreases in growth and photosynthetic parameters occurred under heat stress in WS-6, compared with WS-1. Furthermore, microscopy also revealed greater severity of damage to the photosynthetic apparatus and membrane system of WS-6 under heat stress, compared with WS-1. These effects were probably due to a greater accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA) in WS-6, relative to WS-1. Moreover, higher photochemical efficiency of photosystem II (PSII) and greater activities of antioxidant enzymes were present in WS-1, compared with WS-6. Our results indicated that the extent of heat stress damage in wucai was cultivar dependent. The heat-tolerant cultivar WS-1 had a greater capacity for self-protection and self-regulation under heat stress.

Limited information is available on the cold acclimation of non-heading Chinese cabbage (NHCC) under low temperatures. In this study, the isobaric tags for relative and absolute quantification (iTRAQ) were used to illustrate the molecular machinery of cold acclimation. Compared to the control (Cont), altogether, 89 differentially expressed proteins (DEPs) were identified in wucai leaves responding to low temperatures (LT). Among these proteins, 35 proteins were up-regulated ((and 54 were down-regulated). These differentially expressed proteins were categorized as having roles in carbohydrate metabolism, photosynthesis and energy metabolism, oxidative defense, amino acid metabolism, metabolic progress, cold regulation, methylation progress, and signal transduction. The fructose, glucose, and sucrose were dramatically increased in response to cold acclimation. It was firstly reported that aspartate, serine, glutamate, proline, and threonine were significantly accumulated under low temperatures. Results of quantitative real-time PCR analysis of nine DEPs displayed that the transcriptional expression patterns of six genes were consistent with their protein expression abundance. Our results demonstrated that wucai acclimated to low temperatures through regulating the expression of several crucial proteins. Additionally, carbohydrate and amino acid conversion played indispensable and vital roles in improving cold assimilation in wucai.

Ogura cytoplasmic male sterility (ogu-CMS) has been widely used by breeders for vegetables in the genus Brassica; however, the molecular mechanism of this male sterility remains unclear. The tapetum plays a crucial role in anther development by providing nutrients and secreting callose for pollen development. In this study, we found that the tapetum was abnormally vacuolated after meiosis in Wucai ogu-CMS, which led to the abortion of microspores. The TDF1 gene is essential for early tapetal development. Combined with the characteristics of abortion in ogu-CMS, qRT-PCR analysis showed that the BrTDF1 gene was expressed in all detected tissues from fertile plants, and the transcript levels of this gene in fertile buds were far higher than those in vegetative organs. Compared with the fertile line, BrTDF1 exhibited lower expression in all developmental stages of buds from the sterile line. In addition, the examinations of A6 and EXPB5 genes suggested that the expressions of these genes in ogu-CMS were lower than those in its maintainer line. Further analysis of its function showed that constitutive expression of BrTDF1 in the Arabidopsis tdf1 mutant transgenic lines could restore its fertility. These results indicated that this MYB transcription factor BrTDF1 is required for tapetal development in Wucai.

The effects on root growth, root antioxidant capacity, and cellular ultrastructure were investigated using two wucai genotypes (heat-tolerant WS-1 and heat-sensitive WS-6) under heat stress (40/30 °C) for 5 days. Heat stress caused decreases in root biomass, relative water content (RWC), root vigor, and root traits of two wucai genotypes. In addition, it resulted in reactive oxygen species (ROS) accumulation and increased hydrogen peroxide (H2O2) content, superoxide anion (O2 −) formation rate, and malondialdehyde (MDA) content, but the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were inhibited to different extents in two genotype wucai roots. However, these data indicated that the decline extent of WS-1 (heat tolerant) in root growth and antioxidant capacity was significantly lower than that of WS-6 (Heat sensitive). Microscopic analyses revealed that WS-1 (heat tolerant) showed a better cellular shape than WS-6 under heat stress and slightly oxidative damage; nuclear and mitochondria in WS-1 were of a better intact shape and clear bilayer membrane. Most importantly, the thicker root cell wall in heat-tolerant wucai genotype responding to heat stress was first reported. These results suggested that the ability of heat-tolerant wucai genotype to minimize the heat stress depended upon the higher self-regulation capacity and effectiveness of the antioxidant metabolism.