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Jia Y.,Nanjing Agricultural University | Jia Y.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Jia Y.,Sichuan Agricultural University | Sun J.,Nanjing Agricultural University | And 5 more authors.
Chinese Journal of Applied Ecology | Year: 2011

A water culture experiment was conducted to study the relationship between photosyn-thetic thermal dissipation and xanthophyll cycle in cucumber seedling leaves under hypoxia stress (the dissolved oxygen concentration in nutrient solution was 0.9-1.1 mg.L -1). Under the hypox-ia stress, there was a significant decrease in the quantum yield of PS -photochemistry rate (φPSII), net photosynthetic rate (Pn) under saturation light intensity, quanta yield (AQY), and maximal photochemical efficiency (Fv/Fm ), suggesting that the photoinhibition of the seedling leaves was induced. Meanwhile, the thermal dissipation (NPQ) and the allocation of dissipation energy (D) by antenna increased, but the photochemical quenching apparent (q P) decreased, sug- gesting the enhancement of thermal dissipation in cucumber leaves under hypoxia stress. A positive correlation was observed between NPQ and xanthophyll de-epoxidation state (DEPS), and both of them were promoted by ascorbic acid (AsA) and inhibited by 1,4-dithiothreitol (DTT), suggesting that xanthophyll cycle was the major pathway of photosynthetic thermal dissipation in cucumber seedling leaves under hypoxia stress.

Wang X.H.,Nanjing Agricultural University | Wang X.H.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Song Y.P.,Nanjing Agricultural University | Song Y.P.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | And 10 more authors.
Journal of Animal and Plant Sciences | Year: 2013

Non-heading Chinese cabbage is one of the main vegetables in Asia, especially in China. To analyze the tolerance of non-heading Chinese cabbage to copper (Cu) stress, 10μM, 100μM and 1mM copper were used to treat two cultivars of 'Wutacai' as well as 'Erqing' and then photosynthesis, chlorophyll fluorescence parameters, biomass and chlorophyll content were investigated. In our study, 1mM copper significantly inhibited the photosynthesis and plant growth of both two cultivars. While at 100μM copper, the growth inhibition was just found in 'Erqing', not in 'Wutacai'. The inhibition of copper on the photosynthesis was not only by stomatal factors but also by non-stomatal factors. Copper stress caused damage of photosystem resulting in reduction of PSII efficiency while non-photochemical-quenching parameter NPQ increased. Besides, at low concentration with 10μM, copper promoted the photosynthesis and plant growth. Therefore, the copper tolerance of non-heading Chinese cabbage was dependent on the genotype, and the plant growth inhibition under copper stress was due to the damage of photosystem. Our results will provide a theoretical basis for a further study on non-heading Chinese cabbage tolerance to copper and will be helpful for the production of non-heading Chinese cabbage in heavy metals contaminated areas.

Wang L.,Nanjing Agricultural University | Wang L.,State Key Laboratory of Crop Genetics | Peng H.,Nanjing Agricultural University | Peng H.,State Key Laboratory of Crop Genetics | And 9 more authors.
Plant Biology | Year: 2014

Non-heading Chinese cabbage (Brassica campestris L. ssp. chinensis Makino), an important vegetable crop in China, exhibits a typical sporophytic self-incompatibility (SI) system. To better understand the mechanism of SI response and identify potential candidate proteins involved in the SI system of this vegetable crop, the proteomic approach was taken to identify differential accumulating pistil proteins. Pistils were collected at 0 h and 2 h after self-pollination at anthesis in self-incompatible and compatible lines of non-heading Chinese cabbage, and total proteins were extracted and separated by two-dimensional gel electrophoresis (2-DE). A total of 25 protein spots that displayed differential abundance were identified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF/TOF MS) and peptide mass fingerprinting (PMF). Among them, 22 protein spots were confidently established. The mRNA levels of the corresponding genes were detected by quantitative RT-PCR. The 22 identified protein spots are involved in energy metabolism (four), protein biosynthesis (three), photosynthesis (six), stress response and defence (five), and protein degradation (four). Among these potential candidate proteins, UDP-sugar pyrophosphorylase could be involved in sucrose degradation to influence pollen germination and growth. Glutathione S-transferases could be involved in pollen maturation, and affect pollen fertility. Senescence-associated cysteine protease, which is related to programmed cell death, could be mainly related to self pollen recognition of non-heading Chinese cabbage. The study will contribute to further investigations of molecular mechanism of sporophytic SI in Brassicaceae. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

Ma J.,Nanjing Agricultural University | Ma J.,Longyan University | Hou X.,Nanjing Agricultural University | Hou X.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | And 5 more authors.
Plant Molecular Biology Reporter | Year: 2010

A cDNA clone for the turnip mosaic virus (TuMV)-induced gene in non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino cv. Duanbaigeng) was isolated and characterized. The full-length cDNA clone, designated BcTuR3, was isolated during resistance response to TuMV. Sequence analysis of the cDNA clone confirmed that the translation product of the gene is homologous to other plant resistance proteins. Genomic DNA Southern blot analysis indicated that the gene represented a small multi-gene family. Northern hybridizations confirmed its elevated expression in the resistant "Duanbaigeng" and the susceptible "Aijiaohuang." Upon inoculation with TuMV, the BcTuR3 transcript was rapidly accumulated in the infected leaves and stems. Tissue-specific expression of the gene showed that higher levels of the BcTuR3 transcript were observed in the TuMV-infected leaves, and later in the stems and roots of the resistant and susceptible varieties. These data showed that the BcTuR3 gene may be involved in plant resistance against TuMV pathogen infection. © Springer-Verlag 2010.

Ren J.,Nanjing Agricultural University | Ren J.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Ren J.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Chen Z.,Nanjing Agricultural University | And 20 more authors.
Plant Physiology and Biochemistry | Year: 2013

Ascorbic acid (l-AsA) is an important antioxidant in plants and humans. Vegetables are one of the main sources of ascorbic acid for humans. For instance, non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) is considered as one of the most important vegetables in south China. To elucidate the mechanism by which AsA accumulates, we systematically investigated the expression profiles of d-mannose/. l-galactose pathway-related genes. We also investigated the recycling-related genes and AsA contents in different tissues of three non-heading Chinese cabbage cultivars, 'Suzhouqing', 'Wutacai' and 'Erqing' containing different amounts of AsA. Our results showed that six genes [. d-mannose-6-phosphate isomerase 1 (PMI1), GDP- l-galactose phosphorylase 1 (GGP1), GGP2, GGP4, GDP-mannose-3', 5'-epimerase1 (GME1), and GME2] were expressed at high level and ascorbate oxidase (AAO) was expressed at low level. This expression pattern contributes, at least partially, to higher AsA accumulation in the leaves and petioles than in the roots. Eight genes (PMI1, GME, GGP, l-galactose-1-phosphate phosphatase, l-galactose dehydrogenase, l-galactono-1, 4-lactone dehydrogenase, monodehydroascorbate reductase 1, and glutathione reductase1) were also expressed at high level; AAO and ascorbate peroxidase (APX) were expressed at low level. This expression pattern may similarly contribute to higher AsA accumulation in 'Wutacai' and 'Suzhouqing' than in 'Erqing'. Therefore, the high expression levels of PMI, GME, and GGP and the low expression level of AAO contributed to the high AsA accumulation in non-heading Chinese cabbage. © 2013 Elsevier Masson SAS.

Sun C.,Nanjing Agricultural University | Sun C.,State Key Laboratory of Crop Genetics and oGermplasm Enhancement | Sun C.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Wang L.,Nanjing Agricultural University | And 17 more authors.
Journal of Proteomics | Year: 2014

Downy mildew is a serious fungal disease in non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) that is caused by Hyaloperonospora parasitica, which infects members of the Brassicaceae family. For breeding improvement, researchers must understand the defence mechanisms employed by non-heading Chinese cabbage to combat H. parasitica infection. Using 2-DE protein analysis, we compared the proteomes from leaves of non-heading Chinese cabbage seedlings that were infected with H. parasitica or that were only treated with water at different time points post-infection. By MS analysis, 91 protein spots with significant differences in abundance (>. 2-fold, p<. 0.05) were identified in mock- and H. parasitica-inoculated leaves. Next, a resistance strategy for incompatible interactions was proposed. This network consisted of several functional components, including enhanced ethylene biosynthesis and energy supply, balanced ROS production and scavenging, accelerated protein metabolism and photorespiratory, reduced photosynthesis, and induced photosystem repair. These findings increase our knowledge of incompatible interactions between plants and pathogens and also provide new insight regarding the function of plant molecular processes, which should assist in the discovery of new strategies for pathogen control. Biological significance: This study reported the proteomic analysis of the incompatible interactions between non-heading Chinese cabbage and downy mildew using 2-DE and MS. In total, 91 protein spots that were related to the resistance response were identified. These proteins were assigned to different functional categories, such as amino acid and carbohydrate metabolism, photosynthesis and photorespiration, protein metabolism, signal transduction, redox homeostasis, and ethylene biosynthesis. Meanwhile, several key proteins were determined to be associated with ethylene signalling, ROS scavenging and resistance-related proteins. Consistent with these results, the expression of ethylene biosynthesis genes and response genes, as well as the activity of antioxidant enzymes, increased after inoculation. These findings provide new insight for further understanding the molecular mechanisms of plant resistance. © 2013 Elsevier B.V.

Wang Z.,Nanjing Agricultural University | Wang Z.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Wang Z.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Jiang D.,Nanjing Agricultural University | And 18 more authors.
Gene | Year: 2015

Turnip mosaic virus (TuMV) is the most prevalent viral pathogen infecting most cruciferous plants. MicroRNAs (miRNAs) are around 22 nucleotides long non-protein-coding RNAs that play key regulatory roles in plants. Recent research findings show that miRNAs are involved in plant-virus interaction. However we know little about plant defense and viral offense system networks throughout microRNA regulation pathway. In this study, two small RNA libraries were constructed based on non-heading Chinese cabbage (Brassica campestris ssp. chinensis L. Makino, NHCC) leaves infected by TuMV and healthy leaves, and sequenced using the Illumina-Solexa high-throughput sequencing technology. A total of 86 conserved miRNAs belonging to 25 known miRNA families and 45 novel ones were identified. Among them, twelve conserved and ten new miRNAs were validated by real-time fluorescence quantitative PCR (qPCR). Differential expression analysis showed that 42 miRNAs were down-regulated and 27 miRNAs were up-regulated in response to TuMV stress. A total of 271 target genes were predicted using a bioinformatics approach, these genes are mainly involved in growth and resistance to various stresses. We further selected 13 miRNAs and their corresponding target genes to explore their expression pattern under TuMV and/or cold (4 °C) stresses, and the results indicated that some of the identified miRNAs could link TuMV response with cold response of NHCC. The characterization of these miRNAs could contribute to a better understanding of plant-virus interaction throughout microRNA regulation pathway. This can lead to finding new approach to defend virus infection using miRNA in Chinese cabbage. © 2015.

Wang Z.,Nanjing Agricultural University | Wang Z.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Wang Z.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Wang F.,Nanjing Agricultural University | And 14 more authors.
Acta Physiologiae Plantarum | Year: 2014

C-repeat binding factor (CBF) signaling pathway is involved in cold acclimation responsive to low temperature and some other stresses. CBF transcription factor family is the key component of this pathway. In this study, eight CBF-like genes, BrCBF1, BrCBF2, BrCBF3, BrCBF4, BrCBF5, and BrCBF6A/B/C were isolated from non-heading Chinese cabbage (Brassicacampestris ssp. chinensis L. Makino, NHCC). The deduced CBF proteins shared high similarity with their Arabidopsis orthologs and localized to the nucleus. Furthermore, quantitative real-time PCR (qPCR) analysis showed that BrCBF1~3 were induced by cold (4 °C) but not drought or abscisic acid (ABA), indicating that they are involved in an ABA-independent pathway; however, BrCBF4~6 were regulated by both drought and ABA, suggesting that they were involved in an ABA-dependent pathway. Nevertheless, unlike Arabidopsis, BrCBF4~6 showed response to both cold and ABA, indicates ABA-independent and ABA-dependent parts of CBF pathway in NHCC might not be completely separate, and these genes may act as the connection points in the network. BrCBFs were also accumulated in response to salicylic acid (SA), methyljasmonate (MeJA), and ethylene (ET), indicating that BrCBF genes might participate in the response to biotic stresses. Taken together, eight CBF genes were isolated from NHCC which compose a functional CBF signaling pathway by participating in response to multiple stresses and performing roles from Arabidopsis to some extent. © 2014, Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.

Jiang F.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Jiang F.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Wang F.,State Key Laboratory of Crop Genetics and Germplasm Enhancement | Wang F.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | And 10 more authors.
Plant Molecular Biology Reporter | Year: 2011

Many plants increase in freezing tolerance upon exposure to low non-freezing temperatures, a phenomenon known as cold acclimation. Cold acclimation in Arabidopsis involves rapid cold-induced expression of the inducer of C-repeat/dehydration-responsive element-binding factor (CBF) expression (ICE) transcriptional activators followed by expression of the CBF; subsequently, CBF-targeted genes that increase freezing tolerance. Here, we present evidence for a CBF cold-response pathway in non-heading Chinese cabbage (Brassica campestris ssp. chinensis L. Makino). We show that non-heading Chinese cabbage encodes ICE1-like gene BrICE1 that bracket an open reading frame of 1,491 bp encoding a protein with a potential bHLH domain, which accumulates rapidly in response to low temperature followed closely by expression of the BrCBF gene, an ortholog of the Arabidopsis CBF3-like gene, and then BrCOR14 gene, an ortholog of the Arabidopsis CBF-targeted COR15b gene. An alignment of the later two genes from Arabidopsis, Brassica napus revealed the presence of conserved CANNTG core element and AP2 domain in BrCBF and a CCG core element in BrCOR14. In addition, BrCBF and BrCOR14 showed increased expression induced by low temperature as well as salt and drought, but not by ABA stress which are similar to those of Arabidopsis. We conclude that components of the CBF cold-response pathway are highly conserved in non-heading Chinese cabbage. © 2010 Springer-Verlag.

Jia Y.X.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Sun J.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Guo S.R.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | Li J.,Key Laboratory of Southern Vegetable Crop Genetic Improvement | And 2 more authors.
Russian Journal of Plant Physiology | Year: 2010

The effects of exogenous spermidine (Spd) application to hypoxic nutrient solution on the contents of endogenous polyamines (PAs) and respiratory metabolism in the roots of cucumber (Cucumis sativus L.) seedlings were investigated. Cucumber seedlings were grown hydroponically in control and hypoxic nutrient solutions with and without addition of Spd at a concentration of 0.05 mM. The activities of key enzymes involved in the tricarboxylic acid cycle (TCAC), such as succinate dehydrogenase (SDH) and isocitrate dehydrogenase (IDH), were significantly inhibited under root-zone hypoxia with dissolved oxygen (DO) at 1 mg/l. In contrast, the activities of enzymes involved in the process of fermentation, such as pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and alanine aminotransferase (AlaAT), were significantly increased. Thus, aerobic respiration was inhibited and fermentation was enhanced in the roots of cucumber seedlings as a result of decreasing ATP content to inhibit the dry weight of seedlings under hypoxic stress. Moreover, the contents of free, soluble conjugated, and insoluble bound putrescine (Put), Spd, and spermine (Spm) in the roots of cucumber seedlings were significantly increased under hypoxia stress. Interestingly, application of Spd to hypoxic roots markedly suppressed the accumulation of free Put and, in contrast, promoted an increase in free Spd and Spm, as well as soluble conjugated and insoluble bound Put, Spd, and Spm contents. From these data, we deduced that exogenous Spd promotes the conversion of free Put into free Spd and Spm, and soluble conjugated and insoluble bound PAs under hypoxia stress. Furthermore, the activities of LDH, PDC, and ADH were suppressed and, in contrast, the activities of SDH and IDH were enhanced by application of exogenous Spd to hypoxic roots. As a result, aerobic respiration was enhanced but fermentation metabolism was inhibited in the roots of cucumber seedlings, leading to an increase in ATP content to alleviate the inhibited dry weight of seedlings due to hypoxia stress. These results suggest that application of Spd to hypoxic nutrient solution promoted conversion of free Put into free Spd and Spm as well as soluble conjugated and insoluble bound PAs, further enhanced IDH and SDH activities, and inhibited ethanol fermentation and lactate fermentation, resulting in increased ATP content and eventually enhanced tolerance of cucumber plants to root-zone hypoxia. © 2010 Pleiades Publishing, Ltd.

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