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1. The NAC-like transcription factor SiNAC110 in foxtail millet (Setaria italica L.) confers tolerance to drought and high salt stress through an ABA independent signaling pathway
XIE Li-na, CHEN Ming, MIN Dong-hong, FENG Lu, XU Zhao-shi, ZHOU Yong-bin, XU Dong-bei, LI Lian-cheng, MA You-zhi, ZHANG Xiao-hong
Journal of Integrative Agriculture    2017, 16 (03): 559-571.   DOI: 10.1016/S2095-3119(16)61429-6
摘要1117)      PDF    收藏
Foxtail millet (Setaria italica (L.) P. Beauv) is a naturally stress tolerant crop.  Compared to other gramineous crops, it has relatively stronger drought and lower nutrition stress tolerance traits.  To date, the scope of functional genomics research in foxtail millet (S. italic L.) has been quite limited.  NAC (NAM, ATAF1/2 and CUC2)-like transcription factors are known to be involved in various biological processes, including abiotic stress responses.  In our previous foxtail millet (S. italic L.) RNA seq analysis, we found that the expression of a NAC-like transcription factor, SiNAC110, could be induced by drought stress; additionally, other references have reported that SiNAC110 expression could be induced by abiotic stress.  So, we here selected SiNAC110 for further characterization and functional analysis.  First, the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM (no apical meristem) domain between the 11–139 amino acid positions.  Phylogenetic analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene family.  Subcellular localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis protoplasts.  Gene expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration, high salinity and other abiotic stresses.  Gene functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that, under drought and high salt stress conditions, the seed germination rate, root length, root surface area, fresh weight, and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type (WT), suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt stresses.  However, overexpression of SiNAC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid (ABA) treatment.  Expression analysis of genes involved in proline synthesis, Na+/K+ transport, drought responses, and aqueous transport proteins were higher in the SiNAC110 overexpressed lines than in the WT, whereas expression of ABA-dependent pathway genes did not change.  These results indicated that overexpression of SiNAC110 conferred tolerance to drought and high salt stresses, likely through influencing the regulation of proline biosynthesis, ion homeostasis and osmotic balance.  Therefore SiNAC110 appears to function in the ABA-independent abiotic stress response pathway in plants.
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2. G-protein β subunit AGB1 positively regulates salt stress tolerance in Arabidopsis
MA Ya-nan, CHEN Ming, XU Dong-bei, FANG Guang-ning, WANG Er-hui, GAO Shi-qing, XU Zhao-shi, LI Lian-cheng, ZHANG Xiao-hong, MIN Dong-hong, MA You-zhi
Journal of Integrative Agriculture    2015, 14 (2): 314-325.   DOI: 10.1016/S2095-3119(14)60777-2
摘要1947)      PDF    收藏
The heterotrimeric GTP-binding proteins (G-proteins) in eukaryotes consisted of α, β and γ subunits and are important in molecular signaling by interacting with G-protein-coupled receptors (GPCR), on which to transduce signaling into the cytoplast through appropriate downstream effectors. However, downstream effectors regulated by the G-proteins in plants are currently not well defined. In this study, the transcripts of AGB1, a G protein β subunit gene in Arabidopsis were found to be down-regulated by cold and heat, but up-regulated by high salt stress treatment. AGB1 mutant (agb1-2) was more sensitive to high salt stress than wild-type (WT). Compared with WT, the cotyledon greening rates, fresh weight, root length, seedling germination rates and survival rates decreased more rapidly in agb1-2 along with increasing concentrations of NaCl in normal (MS) medium. Physiological characteristic analysis showed that compared to WT, the contents of chlorophyll, relative proline accumulation and peroxidase (POD) were reduced, whereas the malonaldehyde (MDA) content and concentration ratio of Na+/K+ were increased in agb1-2 under salt stress condition. Further studies on the expression of several stress inducible genes associated with above physiological processes were investigated, and the results revealed that the expressions of genes related to proline biosynthesis, oxidative stress response, Na+ homeostasis, stress- and ABAresponses were lower in agb1-2 than in WT, suggesting that those genes are possible downstream genes of AGB1 and that their changed expression plays an important role in determining phenotypic and physiologic traits in agb1-2. Taken together, these findings indicate that AGB1 positively regulates salt tolerance in Arabidopsis through its modulation of genes transcription related to proline biosynthesis, oxidative stress, ion homeostasis, stress- and ABA-responses.
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3. Characterization of Ethylene Receptors and Their Interactions with GmTPRA Novel Tetratricopeptide Repeat Protein (TPR) in Soybean (Glycine max L.)
NIU Yan-yan, CHEN Ming, XU Zhao-shi, LI Lian-cheng, CHEN Xue-ping , MA You-zhi
Journal of Integrative Agriculture    2013, 12 (4): 571-581.   DOI: 10.1016/S2095-3119(13)60274-9
摘要1701)      PDF    收藏
Ethylene receptors play important roles not only in regulation of growth and development but also in response to environmental stimuli of plants. However, there are few reports on ethylene receptors in soybean. In this article, putative ethylene receptors of soybean were searched from soybean genomic database (http://www.phytozome.net/search.php) and analyzed. The ethylene receptor gene family in soybean comprising eight members, designated as GmERS1-1, GmERS1-2, GmETR1-1, GmETR1-2, GmETR2-1, GmETR2-2, GmEIN4-1, and GmEIN4-2 corresponding with their homologous genes in Arabidopsis, were isolated and analyzed. Phylogenetic analysis indicated that the eight soybean ethylene receptors (SERs) were in two subfamilies and further divided into four groups, viz., groups I (GmERS1-1 and GmERS1-2), II (GmETR1-1 and GmETR1-2), VI (GmETR2-1 and GmETR2-2), and VII (GmEIN4-1 and GmEIN4-2). Protein structure of the members in groups I and II from subfamily I were more conserved than the members in other two groups from subfamily II. Expression patterns of the SERs were compared with the homologous genes in Arabidopsis. The results demonstrated that expression patterns of the SERs differed from Arabidopsis members in the same group, suggesting that SERs are involved in different signal pathways compared to ethylene receptors in Arabidopsis. Promoter analysis showed that the sequences of the members in each group were different from each other, and some specific binding elements of transcription factors detected in promoter sequences might explain the differences between the members in the same group. A novel soybean TPR protein (tetratricopeptide repeat protein), GmTPR, was identified to interact with GmETR1-1, apparently an important ethylene receptor in ethylene signaling pathway in soybean. This suggested that GmTPR might be a novel downstream component of the ethylene signaling pathway.
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4. Isolation and Functional Analysis of the bZIP Transcription Factor Gene TaABP1 from a ChineseWheat Landrace
CAO Xin-you, CHEN Ming*, XU Zhao-shi, CHEN Yao-feng, LI Lian-cheng, YU Yue-hua, LIU Yangna, MA You-zhi
Journal of Integrative Agriculture    2012, 12 (10): 1580-1591.   DOI: 10.1016/S1671-2927(00)8691
摘要1644)      PDF    收藏
In plants, basic leucine zipper (bZIP) transcription factors play important roles in regulatory processes, including stress response, pathogenic defense and light response as well as organ and tissue differentiation. Chinese wheat landrace Pingyaoxiaobaimai (PYXBM), an original parent of drought tolerant wheat varieties grown in northern China, is significantly tolerant to abiotic stresses such as drought, cold and nutrient deficiencies. In order to isolate key stress-responsive genes and then improve stress tolerances of conventional varieties, a bZIP transcription factor gene was isolated from a cDNA library of drought-treated PYXBM using the in situ plaque hybridization method, and was designated as Triticum aestivum L. abscisic acid (ABA)-responsive element binding protein 1 (TaABP1). It encodes 372 amino acids, and contains three conserved domains (C1-C3) in the N terminal and a bZIP domain in the C terminal which is a typical protein structure for the group member of bZIP family. Transcriptional activation analysis showed that TaABP1 activated the expression of downstream reporter genes in yeast without ABA application. TaABP1 protein fused with green fluorescent protein (GFP) demonstrated that the localization of TaABP1 protein is in the nucleus. Expression pattern assays indicated that TaABP1 was strongly induced by ABA, high salt, low temperature and drought, and its expression was stronger in stems and leaves than in the roots of wheat. Furthermore, overexpression of TaABP1 in tobacco showed significant improvement of drought tolerance. Data suggested that TaABP1 may be a good candidate gene for improving stress tolerance of wheat by genetic transformation and elucidation of the role of this gene will be useful for understanding the mechanism underlying drought tolerance of Chinese wheat landrace PYXBM.
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