Abstract Chitinases are pathogensis-related proteins, which play an important role in plant defense mechanisms. The role of the sugarcane chitinase family genes remains unclear due to the highly heterozygous and aneuploidy chromosome genetic background of sugarcane. Ten differentially expressed chitinase genes (belonging to class I~VII) were obtained from RNA-seq analysis of both incompatible and compatible sugarcane genotypes during Sporisorium scitamineum challenge. Their structural properties and expression patterns were analyzed. Seven chitinases ( ScChi I1, ScChi I2, ScChi I3, ScChi III1, ScChi III2, ScChi IV1 and ScChi VI1) showed more positive with early response and maintained increased transcripts in the incompatible interaction than those in the compatible one. Three ( ScChi II1, ScChi V1 and ScChi VII1) seemed to have no significant difference in expression patterns between incompatible and compatible interactions. The ten chitinases were expressed differentially in response to hormone treatment as well as having distinct tissue specificity. ScChi I1, ScChi IV1 and ScChi VII1 were induced by various abiotic stresses (NaCl, CuCl 2 , PEG and 4 °C) and their involvement in plant immunity was demonstrated by over-expression in Nicotiana benthamiana . The results suggest that sugarcane chitinase family exhibit differential responses to biotic and abiotic stress, providing new insights into their function.
Sugarcane smut, which is caused by Sporisorium scitamineum, has been threatening global sugarcane production. Breeding smut resistant sugarcane varieties has been proven to be the most effective method of controlling this particular disease. However, a lack of genome information of sugarcane has hindered the development of genome-assisted resistance breeding programs. Furthermore, the molecular basis of sugarcane response to S. scitamineum infection at the proteome level was incomplete and combining proteomic and transcriptional analysis has not yet been conducted. We identified 273 and 341 differentially expressed proteins in sugarcane smut-resistant (Yacheng05-179) and susceptible (ROC22) genotypes at 48 h after inoculation with S. scitamineum by employing an isobaric tag for relative and absolute quantification (iTRAQ). The proteome quantitative data were then validated by multiple reaction monitoring (MRM). The integrative analysis showed that the correlations between the quantitative proteins and the corresponding genes that was obtained in our previous transcriptome study were poor, which were 0.1502 and 0.2466 in Yacheng05-179 and ROC22, respectively, thereby revealing a post-transcriptional event during Yacheng05-179-S. scitamineum incompatible interaction and ROC22-S. scitamineum compatible interaction. Most differentially expressed proteins were closely related to sugarcane smut resistance such as beta-1,3-glucanase, peroxidase, pathogenesis-related protein 1 (PR1), endo-1,4-beta-xylanase, heat shock protein, and lectin. Ethylene and gibberellic acid pathways, phenylpropanoid metabolism and PRs, such as PR1, PR2, PR5 and PR14, were more active in Yacheng05-179, which suggested of their possible roles in sugarcane smut resistance. However, calcium signaling, reactive oxygen species, nitric oxide, and abscisic acid pathways in Yacheng05-179 were repressed by S. scitamineum and might not be crucial for defense against this particular pathogen. These results indicated complex resistance-related events in sugarcane-S. scitamineum interaction, and provided novel insights into the molecular mechanism underlying the response of sugarcane to S. scitamineum infection.
PDF HTML阅读 XML下载 导出引用 引用提醒 三疣梭子蟹氧化磷酸化代谢在家系近交中的变化 DOI: 作者: 作者单位: 1. 中国水产科学研究院黄海水产研究所, 山东 青岛 266071;2. 青岛海洋科学与技术国家实验室 海洋渔业科学与食物产出过程功能实验室, 山东 青岛 266200 作者简介: 王竹青(1991-),男,硕士研究生,从事甲壳动物遗传育种与繁殖研究.E-mail:leon19910215@outlook.com 通讯作者: 中图分类号: S917 基金项目: 国家自然科学基金项目（41576147，41506186）；泰山领军人才工程高效生态农业创新类计划项目（LJNY2015002）；国家虾蟹产业技术体系项目（CARS-48）. Effects of inbreeding on the oxidative phosphorylation of the swimming crab Portunus trituberculatus Author: Affiliation: 1. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;2. Functional Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:为探究近交对三疣梭子蟹（）氧化磷酸化代谢的影响，首先克隆了三疣梭子蟹线粒体呼吸链4个复合体关键亚基基因的cDNA全长，分别命名为Ndufv2基因cDNA全长1005 bp，编码234个氨基酸，该蛋白是复合体Ⅰ的核心亚基Ndufv2；Cytochrome c1基因全长2371 bp，编码由313个氨基酸组成的亚基Cyt c1；COX6B基因全长1171 bp，编码由105个氨基酸组成的COX6B亚基。生物信息学分析显示，这些复合体亚基基因进化上比较保守。酶活检测及RT-PCR结果表明，随着近交系数的增加，三疣梭子蟹肝胰腺中复合体Ⅰ、复合体Ⅲ活力分别从F4、F2开始呈现显著下降趋势（<0.05）；4个复合体基因表达量均显著下降，且各代表达量显著低于F0（Ndufv2与<0.05），这一结果证实近交衰退已出现在三疣梭子蟹氧化磷酸化代谢通路。 Abstract:The swimming crab (Crustacea:Decapoda:Brachyura) is a dominant species of portunid crab fisheries and an important economic species worldwide. The key performance traits of crustaceans and other aquaculture species are often depressed by inbreeding, especially if inbreeding effects accumulate too rapidly. Although previous studies have investigated the effects of inbreeding on the morphology, hatchability, and survival of crustaceans, little evidence is available that demonstrates that inbreeding affects crustacean physiology. Therefore, in order to investigate the effects of inbreeding on the oxidative phosphorylation of , cDNAs of the key subunit genes of the four mitochondrial respiratory chain complexes were cloned, sequenced using rapid amplification of cDNA ends (RACE), and then analyzed using bioinformatics technology. The full-length cDNA sequence of the Complex I core subunit genes was 1005 bp in length and contained a 705-bp open reading frame (ORF) that encoded a 234-amino acid polypeptide (GenBank:KY682717). The full-length cDNA sequence of Complex Ⅱ was 915 bp in length and contained a 540-bp ORF that encoded a 179-amino acid polypeptide (GenBank:KY406169). The full-length cDNA sequence of the Complex Ⅲ subunit (Cytc1) gene was 2371 bp in length and contained a 942-bp ORF that encoded a 313-amino acid polypeptide (GenBank:KY406171), and the full-length cDNA for the key subunit of Complex IV was 1171 bp in length and encoded 105 amino acids (GenBank:KY406170). In addition, homology and phylogenetic analyses revealed that the amino acid sequences of the four complexes were highly similar to those of closely related species and had higher conservation in evolution, and could be used as a reference for other marine organisms. The activities and mRNA expression of the four complexes in the hepatopancreas and heart mitochondria of were investigated. Results show that inbreeding reduced the activity of all four complexes and their respective subunit gene in the hepatopancreas (<0.05). Besides, Complex I, Ⅲ and IV activities and their subunit genes in heart were declined by inbreeding (<0.05). Furthermore, the elevated activity and expression of Complex Ⅱ in the heart may indicate that the ability to oxidize succinic acid and the level of aerobic metabolism in the crabs rose by breeding might because the dominant homozygous genes were accumulated during the family-based selective breeding programs. Therefore, it is clear that inbreeding gradually reduces the oxidative phosphorylation pathway and provides a reference for family-based selective breeding programs for . 参考文献 相似文献 引证文献
Beta-1,3-glucanases (EC 3.2.1.39), commonly known as pathogenesis-related (PR) proteins, play an important role not only in plant defense against fungal pathogens but also in plant physiological and developmental processes. However, only a limited number of sugarcane beta-1,3-glucanase genes have been isolated. In the present study, we identified and characterized a new beta-1,3-glucanase gene ScGluD2 (GenBank Acc No. KF664181) from sugarcane. An X8 domain was present at the C terminal region of ScGluD2, suggesting beta-1,3-glucan-binding function. Phylogenetic analysis showed that the predicted ScGluD2 protein was classified into subfamily D beta-1,3-glucanase. Localization of the ScGluD2 protein in the plasma membrane was determined by tagging it with green fluorescent protein. The expression of ScGluD2 was more up-regulated in sugarcane smut-resistant cultivars in the early stage (1 d or 3 d) than in the susceptible ones after being challenged by the smut pathogen, revealing that ScGluD2 may be involved in defense against the invasion of Sporisorium scitamineum. Transient overexpression of ScGluD2 in Nicotiana benthamiana leaves induced a defense response and exhibited antimicrobial action on the tobacco pathogens Pseudomonas solanacearum and Botrytis cinerea, further demonstrating that ScGluD2 was related to the resistance to plant pathogens. However, the transcripts of ScGluD2 partially increased (12 h) under NaCl stress, and were steadily up-regulated from 6 h to 24 h upon ABA, H2O2, and CdCl2 treatments, suggesting that ABA may be a signal molecule regulating oxidative stress and play a role in the salt and heavy metal stress-induced stimulation of ScGluD2 transcripts. Taken together, ScGluD2, a novel member of subfamily D beta-1,3-glucanase, was a stress-related gene of sugarcane involved in plant defense against smut pathogen attack and salt and heavy metal stresses.
Heat stress is an increasingly significant abiotic stress factor affecting crop yield and quality. This study aims to uncover the regulatory mechanism of sweet corn response to heat stress by integrating transcriptome and metabolome analyses of seedlings exposed to normal (25 °C) or high temperature (42 °C). The transcriptome results revealed numerous pathways affected by heat stress, especially those related to phenylpropanoid processes and photosynthesis, with 102 and 107 differentially expressed genes (DEGs) identified, respectively, and mostly down-regulated in expression. The metabolome results showed that 12 or 24 h of heat stress significantly affected the abundance of metabolites, with 61 metabolites detected after 12 h and 111 after 24 h, of which 42 metabolites were detected at both time points, including various alkaloids and flavonoids. Scopoletin-7-o-glucoside (scopolin), 3-indolepropionic acid, acetryptine, 5,7-dihydroxy-3′,4′,5′-trimethoxyflavone, and 5,6,7,4′-tetramethoxyflavanone expression levels were mostly up-regulated. A regulatory network was built by analyzing the correlations between gene modules and metabolites, and four hub genes in sweet corn seedlings under heat stress were identified: RNA-dependent RNA polymerase 2 (RDR2), UDP-glucosyltransferase 73C5 (UGT73C5), LOC103633555, and CTC-interacting domain 7 (CID7). These results provide a foundation for improving sweet corn development through biological intervention or genome-level modulation.
Catalases, which consist of multiple structural isoforms, catalyze the decomposition of hydrogen peroxide in cells to prevent membrane lipid peroxidation. In this study, a group II catalase gene ScCAT2 (GenBank Accession No. KF528830) was isolated from sugarcane genotype Yacheng05-179. ScCAT2 encoded a predicted protein of 493 amino acid residues, including a catalase active site signature (FARERIPERVVHARGAS) and a heme-ligand signature (RVFAYADTQ). Subcellular localization experiments showed that the ScCAT2 protein was distributed in the cytoplasm, plasma membrane, and nucleus of Nicotiana benthamiana epidermal cells. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the ScCAT2 gene was ubiquitously expressed in sugarcane tissues, with expression levels from high to low in stem skin, stem pith, roots, buds, and leaves. ScCAT2 mRNA expression was upregulated after treatment with abscisic acid (ABA), sodium chloride (NaCl), polyethylene glycol (PEG), and 4 °C low temperature, but downregulated by salicylic acid (SA), methyl jasmonate (MeJA), and copper chloride (CuCl₂). Moreover, tolerance of Escherichia coli Rosetta cells carrying pET-32a-ScCAT2 was enhanced by NaCl stress, but not by CuCl₂ stress. Sporisorium scitamineum infection of 10 different sugarcane genotypes showed that except for YZ03-258, FN40, and FN39, ScCAT2 transcript abundance in four smut-resistant cultivars (Yacheng05-179, YZ01-1413, YT96-86, and LC05-136) significantly increased at the early stage (1 day post-inoculation), and was decreased or did not change in the two smut-medium-susceptibility cultivars (ROC22 and GT02-467), and one smut-susceptible cultivar (YZ03-103) from 0 to 3 dpi. Meanwhile, the N. benthamiana leaves that transiently overexpressed ScCAT2 exhibited less severe disease symptoms, more intense 3,3'-diaminobenzidine (DAB) staining, and higher expression levels of tobacco immune-related marker genes than the control after inoculation with tobacco pathogen Ralstonia solanacearum or Fusarium solani var. coeruleum. These results indicate that ScCAT2 plays a positive role in immune responses during plant⁻pathogen interactions, as well as in salt, drought, and cold stresses.
Sugarcane smut disease, caused by Sporisorium scitamineum, significantly decreases yield and use of resistant cultivars is the most cost-effective measure for disease control. Current field testing methods for identification of smut resistance are time-consuming and hindered by environmental variability. Our goal was to develop an efficient and reliable resistance identification technique that is rapid, performed in a controlled environment, and stable. Nine sugarcane cultivars with different phenotypic resistance levels were selected. TaqMan quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to measure copy number changes of smut pathogen in sugarcane buds at 0–7 d after needle puncture inoculation. There was a positive correlation between time after inoculation and the amount of smut pathogen in the sugarcane bud. This reached a peak value on 7 d, and the copy number of S. scitamineum decreased in the following order: YZ03-258, FN40, YZ01-1413, GT02-467, ROC22, YT96-86, YZ03-103, FN39, LC05-136. After smut pathogen inoculation, differences in the physiological and biochemical indices of the nine cultivars were observed. Peroxidase (POD), ascorbate peroxidase (APX), catalase (CAT), superoxide dismutase (SOD), β-1,3-glucanase, and malondialdehyde (MDA) were grouped into three main components, and the cumulative contribution rate was 80.177%, revealing that these are useful physiological and biochemical indicators of smut resistance. Subordinate function analysis indicated that the levels of smut resistance of the nine genotypes were (high to low): YZ03-258, FN40, YZ01-1413, GT02-467, ROC22, YZ03-103, YT96-86, FN39, LC05-136, which is similar to the results from copy number determination of smut pathogens. The results suggest that after artificial needle inoculation, rapid identification of physiological resistance to sugarcane smut was achieved based on copy number increases in the sugarcane smut pathogen and the physiological and biochemical changes in sugarcane bud during the early phase of infection.
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