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.

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.

The variation among Chinese genotypes of Brassica napus L. for seed tocopherols content and their analysis using gas chromatography has not been comprehensively reported till to date. In the present study, the tocopherol contents of four Chinese genotypes of Brassica napus L., namely, Gaoyou 605, Zhejiang 619, Zheshuang 758, and Zheshuang 72, were evaluated using three modified sample preparation protocols (P1, P2, and P3) for tocopherol extraction. These methods were distinguished as follows. Protocol one (P1) included the evaporation of solvent after extraction without silylation. Protocol two (P2) followed the direct supernatant collection after overnight extraction without drying and silylation. Protocol three (P3) included trimethylsilylation with N,O-bis(trimethylsilyl) trifluoroacetamide. Genotypic comparison of tocopherol and its isoforms revealed that Gaoyou 605 was dominant over the other genotypes with (140.5±10.5), (316.2± 9.2), and (559.1± 24.3) μg g-1 of seed meal α-, γ-, and total (T-) tocopherol, respectively, and a 0.44±0.04 α- to γ-tocopherol ratio. The comparison of the sample preparation protocols, on the other hand, suggests that P3 is the most suitable method for the tocopherol extraction from Brassica oilseeds and for the analysis of tocopherols using gas chromatography flame ionization detector (GC-FID). Trimethylsilylation is the key step differentiating P3 from P1 and P2. Variations detected in tocopherol contents among the Chinese rapeseed (B. napus) genotypes signify the need to quantify a wide range of rapeseed germplasm for seed tocopherol dynamics in short and crop improvement in long.

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.

Phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) catalyses phosphoenolpyruvate (PEP) to yield oxaloacetate, which is involved in protein biosynthesis. Pyruvate kinase (PK; EC 2.7.1.40) catalyzes PEP to yield pyruvate, which is involved in fatty acid synthesis. In this study, five PEPC genes (AhPEPC1, AhPEPC2, AhPEPC3, AhPEPC4, and AhPEPC5) from peanut have been cloned. Using a quantitative real-time RT-PCR approach, the expression pattern of each gene was monitored during the seed development of four peanut varieties (E11, Hebeigaoyou, Naihan 1, and Huayu 26). It was found that these five genes shared similar expression behaviors over the developmental stages of E11 with high expression levels at 30 and 40 d after pegging (DAP); whereas these five genes showed irregular expression patterns during the seed development of Hebeigaoyou. In Naihan 1 and Huayu 26, the expression levels of the five genes remained relatively high in the first stage. The PEPC activity was monitored during the seed development of four peanut varieties and seed oil content was also characterized during whole period of seed development. The PEPC activity followed the oil accumulation pattern during the early stages of development but they showed a significantly negative correlation thereafter. These results suggested that PEPC may play an important role in lipid accumulation during the seed development of four peanut varieties tested.

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.