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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

Background Chlorophyll (Chl) is a vital photosynthetic pigment involved in capturing light energy and energy conversion. In this study, the color conversion of inner-leaves from green to yellow in the new wucai (Brassica campestris L.) cultivar W7–2 was detected under low temperature. The W7–2 displayed a normal green leaf phenotype at the seedling stage, but the inner leaves gradually turned yellow when the temperature was decreased to 10 °C/2 °C (day/night), This study facilitates us to understand the physiological and molecular mechanisms underlying leaf color changes in response to low temperature. Results A comparative leaf transcriptome analysis of W7–2 under low temperature treatment was performed on three stages (before, during and after leaf color change) with leaves that did not change color under normal temperature at the same period as a control. A total of 67,826 differentially expressed genes (DEGs) were identified. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) analysis revealed that the DEGs were mainly enriched in porphyrin and Chl metabolism, carotenoids metabolism, photosynthesis, and circadian rhythm. In the porphyrin and chlorophyll metabolic pathways, the expression of several genes was reduced [i.e. magnesium chelatase subunit H (CHLH)] under low temperature. Almost all genes [i.e. phytoene synthase (PSY)] in the carotenoids (Car) biosynthesis pathway were downregulated under low temperature. The genes associated with photosynthesis [i.e. photosystem II oxygen-evolving enhancer protein 1 (PsbO)] were also downregulated under LT. Our study also showed that elongated hypocotyl5 (HY5), which participates in circadian rhythm, and the metabolism of Chl and Car, is responsible for the regulation of leaf color change and cold tolerance in W7–2. Conclusions The color of inner-leaves was changed from green to yellow under low temperature in temperature-sensitive mutant W7–2. Physiological, biochemical and transcriptomic studies showed that HY5 transcription factor and the downstream genes such as CHLH and PSY, which regulate the accumulation of different pigments, are required for the modulation of leaf color change in wucai under low temperature.

Background The discovery of male sterile materials is of great significance for the development of plant fertility research. Wucai (Brassica campestris L. ssp. chinensis var. rosularis Tsen) is a variety of non-heading Chinese cabbage. There are few studies on the male sterility of wucai, and the mechanism of male sterility is not clear. In this study, the male sterile mutant MS7–2 and the wild-type fertile plant MF7–2 were studied. Results Phenotypic characteristics and cytological analysis showed that MS7–2 abortion occurred at the tetrad period. The content of related sugars in the flower buds of MS7–2 was significantly lower than that of MF7–2, and a large amount of reactive oxygen species (ROS) was accumulated. Through transcriptome sequencing of MS7–2 and MF7–2 flower buds at three different developmental stages (a–c), 2865, 3847, and 4981 differentially expressed genes were identified in MS7–2 at the flower bud development stage, stage c, and stage e, respectively, compared with MF7–2. Many of these genes were enriched in carbohydrate metabolism, phenylpropanoid metabolism, and oxidative phosphorylation, and most of them were down-regulated in MS7–2. The down-regulation of genes involved in carbohydrate and secondary metabolite synthesis as well as the accumulation of ROS in MS7–2 led to pollen abortion in MS7–2. Conclusions This study helps elucidate the mechanism of anther abortion in wucai, providing a basis for further research on the molecular regulatory mechanisms of male sterility and the screening and cloning of key genes in wucai.

Background Wucai (Brassica campestris L. ssp. chinensis var. rosularis Tsen) is a cold-tolerant plant that is vulnerable to high temperature. This study explored the response mechanism of wucai to low temperature. In this study, wucai seedlings were treated with different temperatures, including low temperature (LT), high temperature (HT), and a control. Results According to transcriptomics analysis, the number of differentially expressed genes (DEGs) in HT and LT was 10,702 and 7267, respectively, compared with the control. The key genes associated with the physiological response of wucai to the treatments were analyzed. The Kyoto Encyclopedia of Genes and Genomes and Gene Ontology annotations indicated the importance of the photosynthesis and photosynthetic-antenna protein pathways. We found that a high-temperature environment greatly inhibited the expression of important genes in the photosynthetic pathway (BrLhc superfamily members, PsaD, PsaE, PsaD, PsaD, PsbO, PsbP, PsbQ, PsbR, PsbS, PsbW, PsbY, Psb27, and Psb28), whereas low temperature resulted in the expression of certain key genes (BrLhc superfamily members, Psa F, Psa H, Psb S, Psb H, Psb 28). In addition, the wucai seedlings exhibited better photosynthetic performance under low-temperature conditions than high-temperature conditions. Conclusions Based on the above results, we speculate that upon exposure to low temperature, the plants developed higher cold tolerance by upregulating the expression of genes related to photosynthesis. Conversely, high-temperature stress inhibited the expression of pivotal genes and weakened the self-regulating ability of the plants.

Background Leaf color mutants are the ideal materials to explore the pathways of chlorophyll (Chl) metabolism, chloroplast development, and photosynthesis system. In this study, a spontaneous yellow-green leaf wucai (Brassica campestris L.) mutant “WY16–13” was identified, which exhibited yellow-green leaf color during its entire growth period. However, current understanding of the molecular mechanism underlying Chl metabolism and chloroplast development of “WY16–13” is limited. Results Total Chl and carotenoid content in WY16–13 was reduced by 60.92 and 58.82%, respectively, as compared with its wild type parental line W16–13. Electron microscopic investigation revealed fewer chloroplasts per cell and looser stroma lamellae in WY16–13 than in W16–13. A comparative transcriptome profiling was performed using leaves from the yellow-green leaf type (WY16–13) and normal green-leaf type (W16–13). A total of 54.12 million (M) (WY16–13) and 56.17 M (W16–13) reads were generated. A total of 40,578 genes were identified from the mapped libraries. We identified 3882 differentially expressed genes (DEGs) in WY16–13 compared with W16–13 (i.e., 1603 upregulated genes and 2279 downregulated genes). According to the Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, these DEGs are involved in porphyrin and Chl metabolism [i.e., chlorophyllase (CLH), heme oxygenase (HO), chlorophyll (ide) b reductase (NYC), and protochlorophyllide oxidoreductase (POR) genes], carbohydrate metabolism, photosynthesis, and carbon fixation in photosynthetic organisms. Moreover, deficiency in Chl biosynthetic intermediates in WY16–13 revealed that the formation of the yellow-green phenotype was related to the disorder of heme metabolism. Conclusions Our results provide valuable insights into Chl deficiency in the yellow-green leaf mutant and a bioinformatics resource for further functional identification of key allelic genes responsible for differences in Chl content.

The novel male sterility line 12-14A of Wucai (Brassica campestris) was generated by backcrossing. In this study, morphological, molecular, and cytological analyses were performed to characterize this line. The phenotypes of flower organs of 12-14A were different from those of its maintainer line 12-14B, as the flowers of 12-14A were completely infertile. Molecular analysis confirmed that the male sterility line 12-14A was classified as the ogu CMS type. An analysis of anther development in the CMS line demonstrated that two types of abortion occur simultaneously: pollen mother cell abortion and tetrad-to-mononuclear microspore abortion. In addition, the mature anthers of 12-14A showed typical abortive features, such as appearing dry, shrunken, and scaly. A heterosis test of hybrids from CMS 12-14A and four inbred lines was carried out, and the F1 hybrids exhibited clear heterosis in plant weight, as well as in the protein, soluble sugar, and vitamin C content, suggesting high yields and high quality potential. These results indicate that this novel CMS line may have powerful potential in Wucai hybrid breeding.

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.

Background: Auxin signalling plays a crucial role in plant growth and development. Although the auxin response factor (ARF) gene family has been studied in some plant species, its structural features, molecular evolution, and expression profile in Zizania latifolia (Z. latifolia) are still not clear.Results: Our study identified 33 putative YSL genes from the whole Z. latifolia genome. Furthermore, a comprehensive overview of the ZlARFs was undertaken, including phylogenetic relationship, gene structures, conserved domains, synteny, Ka/Ks, motifs, and subcellular locations of the gene product. Synteny analyses and the calculation of Ka/Ks values suggested that all 57 orthologous/paralogous gene pairs between Z. latifolia and Z. latifolia, Z. latifolia and Oryza sativa have experienced strong purifying selection. The phylogenetic analysis of ARFs indicated that the ZlARFs can be divided into 6 classes and that most ZlARFs from Z. latifolia have closer relationships with Oryza sativa than with Arabidopsis. RNA-Seq data and qRT-PCR analyses showed that ZlARF genes were expressed in TDF treatment and U. esculenta infection, while some ZlARFs exhibited high expression levels only in U. esculenta infection. Meanwhile, the interaction networks and gene ontology (GO) term of the ZlARF genes were constructed and 23 ZlARF co-expressed genes were identified, most of which were down-regulated involve auxin-activated signalling pathway in after swelling stem formation. Transcriptome analysis results verified the relevant functions of ARF genes, and most ZlARF genes regulated physiological processes in response to differential cell expansion. Conclusion: Comprehensive bioinformation analysis of the auxin response factor gene family (ARF) in Z. latifolia and its association with swelling stem formation after U. esculenta infection. The bioinformatic and RNA-Seq analyses provided valuable information for further study on the regulation of the growth and development of swelling stem formation by ZlARFs in Z. latifolia.

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.

Background Brassinosteroids (BRs) have a positive effect on many processes during plant growth and development, and in response to various abiotic stressors. Low-temperature (LT) stress constricts the geographic distribution, growth, and development of wucai (Brassica campestris L. ssp. chinensis var. rosularis Tsen). However, there is little information on the global gene expression of BRs under LT stress in wucai. In this study, the molecular roles of 24-epibrassinolide (EBR) after exogenously application, were explored by RNA sequencing under LT conditions. Results According to the Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, photosynthesis was significantly enriched after spraying EBR under LT. The transcripts encoding the photosystem II (PSII) oxygen-evolving enhancer protein, photosystem I (PSI) subunit, light-harvesting chlorophyll protein complexes I and II, and ferredoxin were up-regulated after the application of EBR. Transcripts encoding several key enzymes involved in chlorophyll biosynthesis were also up-regulated, accompanied by significant differences in the contents of 5-aminolevulinic acid (ALA), porphobilinogen (PBG), protoporphyrin IX (Proto IX), Mg-protoporphyrin IX (Mg-proto IX), protochlorophyllide (Pchl), and photosynthetic pigments. Notably, transcriptional and physiological analyses revealed that under LT stress, plant responses to EBR involved a major reorientation of photosynthesis, as well as porphyrin and chlorophyll metabolism. Conclusion This study explored the role of EBR as an LT stress tolerance mechanism in wucai. At the transcription level, LT tolerance manifests as an enhancement of photosynthesis, and the amelioration of porphyrin and chlorophyll metabolism.

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.

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.

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.

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.

Chlorophyll is a vital photosynthetic pigment that plays a key role in plant development, participating in light energy capture and energy conversion. In this study, a novel wucai ( Brassica campestris L.) germplasm with green outer leaves and yellow inner leaves at the adult stage (W7-2) was used to examine chlorophyll metabolism response to cold acclimation. A green leaf wucai genotype without leaf color changes named W7-1 was selected as the control to evaluate the chlorophyll metabolism changes of W7-2. Compared to W7-1, the contents of chlorophyll a (Chl a) and chlorophyll b (Chl b) in W7-2 were significantly reduced at five developmental stages (13, 21, 29, 37, and 45 days after planting (DAP)). An iTRAQ-based quantitative proteomic analysis was carried out at 21 and 29 DAP according to the leaf color changes in both of genotypes. 1409 proteins were identified, while 218 of them displayed differential accumulations between W7-2 and W7-1 during the two developmental stages. The differentially expressed proteins (DEPs) mainly assigned to chlorophyll biosynthesis, photosynthesis, carbohydrate metabolism, ribosome metabolism and posttranslational modification. Among these DEPs, NADPH-protochlorophyllide oxidoreductase (PORB) and Mg-protoporphyrin IX chelatase 1 (CHLI1) were the key enzymes participating in chlorophyll (Chl) biosynthesis, which was down-regulated at 21 DAP and up-regulated at 29 DAP in W7-2 compared with W7-1, respectively. The expression analysis of genes of three subunits of Mg-chelatase ( CHLI1, CHLD, and CHLH), Genomes Uncoupled 4 ( GUN4), and Thioredoxin ( TRX3) associated with chlorophyll metabolism also displayed significant down-regulation in W7-2. In particular, PORB showed significant up-regulation in W7-2, significantly affecting chlorophyll biosynthesis. Additionally, differences in chlorophyll metabolism between W7-2 and W7-1 were in terms of altered photosynthesis, carbohydrate, and energy metabolism. We found that the transcription levels of most photosynthesis proteins showed significantly lower levels, and the genes expression level, associated with carbohydrate and energy metabolism, were lower in W7-2 than in W7-1. Therefore, the present study results help understand the physiological and molecular mechanisms underlying leaf coloring responding to cold acclimation.

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].

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.

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.

Myo-inositol is a vital compound in plants. As the key rate-limiting enzyme in myo-inositol biosynthesis, l-myo-inositol-1-phosphate synthase (MIPS) is regarded as a determinant of the myo-inositol content in plants. The up-regulation of MIPS genes can increase the myo-inositol content, thereby enhancing the plant's resistance to a variety of stresses. However, there are few reports on the roles of myo-inositol and the identification of MIPS in woody trees. In this study, a MIPS gene, named as PeMIPS1, was characterized from Populus euphratica Oliv. The heterologous expression of PeMIPS1 compensated for inositol production in the yeast inositol auxotrophic mutant ino1 and the phenotypic lesions of the atmips1-2 mutant, an Arabidopsis MIPS1 knock-out mutant. A subcellular location analysis showed that the PeMIPS1-GFP fusion was localized in the nucleus and cytoplasm, but not in the chloroplasts, indicating that PeMIPS1 represented the cytosolic form of MIPS in P. euphratica. Interestingly, PeMIPS1 was not only inducible by drought and high salinity, but also by CuSO4 treatment. The transgenic poplar lines overexpressing PeMIPS1 had greater plant heights, shoot biomasses and survival rates than the wild type during the salt- or copper-stress treatment, and this was accompanied by an increase in the myo-inositol content. The overexpression of PeMIPS1 resulted in the increased activities of antioxidant enzymes and the accumulation of ascorbate, a key nonenzymatic antioxidant in plant, which partly accounted for the enhanced reactive oxygen species-scavenging capacity and the lowered hydrogen peroxide and malondialdehyde levels in the transgenic poplar. To the best of our knowledge, this study is the first to report the roles of MIPS genes in the tolerance to copper stress.

Heat stress is a major environmental stress that limits plant growth and yield worldwide. The present study was carried out to explore the physiological mechanism of heat tolerant to provide the theoretical basis for heat-tolerant breeding. The changes of leaf morphology, anatomy, nitrogen assimilation, and carbohydrate metabolism in two wucai genotypes (WS-1, heat tolerant; WS-6, heat sensitive) grown under heat stress (40°C/30°C) for 7 days were investigated. Our results showed that heat stress hampered the plant growth and biomass accumulation in certain extent in WS-1 and WS-6. However, the inhibition extent of WS-1 was significantly smaller than WS-6. Thickness of leaf lamina, upper epidermis, and palisade mesophyll were increased by heat in WS-1, which might be contributed to the higher assimilation of photosynthates. During nitrogen assimilation, WS-1 possessed the higher nitrogen-related metabolic enzyme activities, including nitrate reductase (NR), glutamine synthetase (GS), glutamate synthase (GOGAT), and glutamate dehydrogenase (GDH), which were reflected by higher photosynthetic nitrogen-use efficiency (PNUE) with respect to WS-6. The total amino acids level had no influence in WS-1, whereas it was reduced in WS-6 by heat. And the proline contents of both wucai genotypes were all increased to respond the heat stress. Additionally, among all treatments, the total soluble sugar content of WS-1 by heat got the highest level, including higher contents of sucrose, fructose, and starch than those of WS-6. Moreover, the metabolism efficiency of sucrose to starch in WS-1 was greater than WS-6 under heat stress, proved by higher activities of sucrose phosphate synthase (SPS), sucrose synthase (SuSy), acid invertase (AI), and amylase. These results demonstrated that leaf anatomical alterations resulted in higher nitrogen and carbon assimilation in heat-tolerant genotype WS-1, which exhibited a greater performance to resist heat stress.

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.

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.