Growth and productivity of rice (Oryza sativa L.) are severely affected by salinity. Understanding the mechanisms that protect rice and other important cereal crops from salt stress will help in the development of salt-stress-tolerant strains. In this study, rice seedlings of the same genetic species with various salt tolerances were studied. We first used 2DE to resolve the expressed proteome in rice roots and leaves and then used nanospray liquid chromatography/tandem mass spectrometry to identify the differentially expressed proteins in rice seedlings after salt treatment. The 2DE assays revealed that there were 104 differentially expressed protein spots in rice roots and 59 in leaves. Then, we identified 83 proteins in rice roots and 61 proteins in rice leaves by MS analysis. Functional classification analysis revealed that the differentially expressed proteins from roots could be classified into 18 functional categories while those from leaves could be classified into 11 functional categories. The proteins from rice seedlings that most significantly contributed to a protective effect against increased salinity were cysteine synthase, adenosine triphosphate synthase, quercetin 3-O-methyltransferase 1, and lipoxygenase 2. Further analysis demonstrated that the primary mechanisms underlying the ability of rice seedlings to tolerate salt stress were glycolysis, purine metabolism, and photosynthesis. Thus, we suggest that differentially expressed proteins may serve as marker group for the salt tolerance of rice.
Our results not only provide a comprehensive overview of the starch biosynthetic pathway in the developing endosperm but also reveal some important protein markers that regulate the synthesis of starch. In human diets, rice (Oryza sativa L.) is an important source of starch, a substantial amount of which is accumulated in developing endosperm. A better understanding of the complicated pathways involved in starch biosynthesis is needed to improve the yield and quality of rice and other cereal crops through breeding. One pure line rice mutant, SA0419, was induced from a wild-type rice, TNG67, by sodium azide mutagenesis; therefore, TNG67 and SA0419 share the same genetic background. SA0419 is, however, a unique glutinous rice with a lower amylose content (8%) than that of TNG67 (20%), and the grains of SA0419 develop earlier and faster than those of TNG67. In this study, we used a comparative proteomic analysis to identify the differentially expressed proteins that may explain the differences in starch biosynthesis and the characteristics of TNG67 and SA0419. A gel-based proteomic approach was applied to profile the expressed proteome in the developing endosperm of these two rice varieties by nano-LC/MS/MS. Several over-expressed proteins were found in SA0419, such as plastidial ADP-glucose pyrophosphorylase (AGPase), phosphoglucomutase (PGM), pyrophosphate-fructose 6-phosphate 1-phosphotransferase (PFP), 6-phosphofructokinase (PFK), pyruvate phosphate dikinase (PPDK), starch branching enzymes (SBE) and starch debranching enzyme (SDBE), with those proteins mainly being involved in the pathways of starch metabolism and PPDK-mediated gluconeogenesis. Those over-expressed enzymes may contribute to the relatively early development, similar starch accumulation and rapid grain filling of SA0419 as compared with TNG67. This study provides a detailed biochemical description of starch biosynthesis and related information regarding a unique starch mutant that may assist future research efforts to improve the yield and quality of grain and starch in rice through breeding.
SUMMARYFusarium wilt of watermelon, caused by Fusarium oxysporum f. sp. niveum, is one of the limiting factors for watermelon production in Taiwan. In recent research, the phenylalanine ammonia lyase (PAL) gene expressed in the shoot base of the Fusarium wilt resistant line JSB was related to Fusarium wilt resistance. Phenylalanine ammonia lyase is the key regulatory enzyme in the phenylpropanoid metabolic pathway. The downstream products of phenolic compounds are considered to be involved in the complicated plant defence mechanisms. They could act as signal molecules, antimicrobial substances and/or structural barriers. To study the resistant mechanisms of Fusarium wilt, the resistant JSB line was examined for comparison of F. oxysporum-watermelon interactions with the susceptible Grand Baby (GB) cultivar. Unlike infected GB, which was seriously colonized by F. oxysporum in the whole plant, the pathogen was limited below the shoot base of inoculated JSB, suggesting that the shoot base of JSB may contribute to Fusarium resistance. The data indicated that a significant increase in PAL activity was found in shoot bases of the resistant JSB line at 3, 9, 12 and 15 days after inoculation (DAI). Shoot bases of resistant watermelons accumulated higher amounts of soluble and cell wall-bound phenolics at 3–9 DAI; the susceptible GB cultivar, however, only increased the cell wall-bound phenolics in shoot bases at 3 DAI. High lignin deposition in the cell walls of vascular bundles was observed in the shoot bases of JSB but not of GB seedlings at 6 and 9 DAI. In the roots and shoot bases of JSB seedlings at 6 DAI, peroxidase enzyme activity increased significantly. In summary, the results suggest that accumulation of cell wall-bound phenolics and increase of peroxidase activity in shoot bases of JSB seedlings during F. oxysporum inoculation, together with the rapid deposition of lignin in the cell walls of vascular bundles, may have provided structural barriers in resistant JSB line to defend against F. oxysporum invasion.
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