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Leaf senescence is a notably important trait that limits the yield and biomass accumulation of agronomic crops. Therefore, determining the chromosomal position of the expression sequence tags (ESTs) that are associated with leaf senescence is notably interesting in the manipulation of leaf senescence for crop improvement. A total of 32 ESTs that were previously identified during the delaying leaf senescence stage in the stay-green wheat cultivar CN17 were mapped to 42 chromosomes, a chloroplast, a mitochondrion, and a ribosome using in silico mapping. Then, we developed 19 pairs of primers based on these sequences and used them to determine the polymorphisms between the stay-green cultivars (CN12, CN17, and CN18) and the control cultivar MY11. Among the 19 pairs of primers, 5 pairs produced polymorphisms between the stay-green cultivar and the non-stay-green control. Further studies of Chinese Spring nullisomic-tetrasomics show that JK738991 is mapped to 3B, JK738983 is mapped to 5D, and JK738989 is mapped to 2A, 4A, and 3D. The other two ESTs, JK738994 and JK739003, were not assigned to a chromosome using the Chinese Spring nullisomic-tetrasomics, which indicates that these ESTs may be derived from rye DNA in the wide cross. In particular, the ESTs that produce polymorphisms are notably useful in identifying the stay-green cultivar using molecular marker-assisted selection. The results also suggest that the in silico mapping data, even from a comparison genomic analysis based on the homogeneous comparison, are useful at some points, but the data were not always reliable, which requires further investigation using experimental methods.

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Thinopyrum ponticum (2n = 10x = 70) has donated rust resistance genes to protect wheat from this fungal disease. In the present study, the line ES-7, derived from the progeny of the crosses between common wheat cultivar Abbondanza and Triticum aestivumTh. ponticum partial amphiploid line Xiaoyan784, was characterized by cytological, fluorescence in situ hybridization (FISH), genomic in situ hybridization (GISH) and EST-STS marker techniques. Cytological observations revealed that the configuration of ES-7 was 2n = 42 = 21 II. GISH and FISH results showed that ES-7 had two St chromosomes and lacked 5A chromosomes compared to common wheat. The 4A chromosome of ES-7 had small alterations from common wheat. Two EST-SSR markers BE482522 and BG262826, specific to Th. ponticum and tetraploid Pseudoroegneria spicata (2n = 4x = 28), locate on the homoeologous group 5 chromosomes of wheat, could amplify polymorphic bands in ES-7. It was suggested that the introduced St chromosomes belonged to homoeologous group 5, that is, ES-7 was a 5St (5A) disomic substitution line. Furthermore, ES-7 showed highly resistance to mixed stripe rust races of CYR32 and CYR33 in adult stages, which was possibly inherited from Th. ponticum. Thus, ES-7 can be used for wheat stripe rust resistance breeding program.

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The forward and reverse cDNA subtractive libraries before and after the toxic effect of α-amanitin were constructed by suppression subtractive hybridization and randomly selected clones from each subtractive library were screened by PCR and dot blot hybridization. A total of 85 genes with altered expression were finally identified, with 41 genes from the forward library and 44 from the reverse library. Subsequently, the antagonistic effects of candidate traditional Chinese medicines were evaluated based on the genetic transcription levels of the genes with significant altered expression, including Catnβ, Flt3-L, IL-7r and Rpo2-4. The results indicated that Silybum marianum (L.) Gaert and Ganoderma lucidum had significant down-regulated effects on the transcription level of Catnβ that was up-regulated by α-amanitin, and the two herbs also up-regulated the transcription levels of Flt3-L and Rpo2-4. Silybum marianum (L.) had significant up-regulated effects on the IL-7r that was down-regulated by α-amanitin. These preliminary studies suggested that Silybum marianum (L.) and Ganoderma lucidum were effective antagonists against the toxicity of α-amanitin.

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Gibberellins (GAs) are a class of plant hormones that play important roles in diverse aspects during plant growth and development. A series of GA synthesis and metabolism genes have been reported or proved to have essential functions in different plant species, while a small number of GA 2-oxidase genes have been cloned or reported in wheat. Previous studies have provided some important findings on the process of GA biosynthesis and the enzymes involved in its related pathways. These may facilitate understanding of the complicated process underlying GA synthesis and metabolism in wheat. In this study, GA 2-oxidase genes TaGA2ox1-1, TaGA2ox1-2, TaGA2ox1-3, TaGA2ox1-4, TaGA2ox1-5, and TaGA2ox1-6 were identified and further overexpressed in rice plants to investigate their functions in GA biosynthesis and signaling pathway. Results showed overexpression of GA 2-oxidase genes in rice disrupted the GA metabolic pathways and induced catalytic responses and regulated other GA biosynthesis and signaling pathway genes, which further leading to GA signaling disorders and diversity in phenotypic changes in rice plants.

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Acta Alimentaria
Authors:
Y.L. Xu
,
Y.D. Zhang
,
Z.P. Wang
,
W.W. Chen
,
C. Fan
,
J.Q. Xu
,
T. Wang
, and
S. Rong

Abstract

To explore the effect of sesamol on the cognition of APP/PS1 mice, 8-week-old APP/PS1 and wild-type male mice were divided into AD model group, AD + sesamol (50 mg kg−1 bw) group, and Control group. Sesamol was orally administered once a day for 5 months. Morris water maze was used to evaluate the learning and memory ability of mice. The number of synapses in the hippocampal neurons was detected by Golgi staining. Nissl staining was used to observe the changes of Nissl bodies in CA1 and CA3 regions of the hippocampus. Western blotting was used to detect the expression of Aβ, SIRT1, BDNF, and p-CREB/CREB in the hippocampus and cortex. Compared with the model group, sesamol decreased the latency period of APP/PS1 mice (P < 0.05) and increased the total number of neuronal dendritic spines in the hippocampal CA3 region, as well as increased the number of Nissl bodies (P < 0.05). Western blotting results showed that sesamol significantly reduced Aβ protein expression in the hippocampus and cortex, increased SIRT1 expression in the cortex, and increased BDNF expression in the hippocampus (P < 0.05). Sesamol improved the learning and memory abilities of APP/PS1 mice probably through increasing the density of neuronal dendritic spines and upregulating the levels of SIRT1 and BDNF.

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Chinese endemic wheat landraces possess unique morphological features and desirable traits, useful for wheat breeding. It is important to clarify the relationship among these landraces. In this study, 21 accessions of the four Chinese endemic wheat landrace species were investigated using single-copy genes encoding plastid Acetyl-CoA carboxylase (Acc-1) and 3-phosphoglycerate kinase (Pgk-1) in order to estimate their phylogenetic relationship. Phylogenetic trees were constructed using maximum parsimony (MP), maximum likelihood (ML) and Bayesian, and TCS network and gene flow values. The A and B genome sequences from the Pgk-1 loci indicated that three accessions of Triticum petropavlovskyi were clustered into the same subclade, and the T. aestivum ssp. tibetanum and the Sichuan white wheat accessions were grouped into a separate subclade. Based on the Acc-1 gene, T. aestivum ssp. tibetanum and T. aestivum ssp. yunnanense were grouped into one subclade in the A genome; the B genome from T. petropavlovskyi and T. aestivum ssp. tibetanum, and the Sichuan white wheat complex and T. aestivum ssp. tibetanum were grouped in the same clades. The D genome of T. aestivum ssp. yunnanense clustered with T. petropavlovskyi. Our findings suggested that (1) T. petropavlovskyi is distantly related to the Sichuan white wheat complex; (2) T. petropavlovskyi, T. aestivum ssp. tibetanum and T. aestivum ssp. yunnanense are closely related; (3) T. aestivum ssp. tibetanum is closely related to T. aestivum ssp. yunnanense and the Sichuan white wheat complex; and (4) T. aestivum ssp. tibetanum may be an ancestor of Chinese endemic wheat landraces.

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In this study, the cDNA of homocysteine S-methyltransferase was isolated from Aegilops tauschii Coss., with the gene accordingly designated as AetHMT1. Similar to other methyltransferases, AetHMT1 contains a GGCCR consensus sequence for a possible zinc-binding motif near the C-terminal and a conserved cysteine residue upstream of the zinc-binding motif. Analysis of AetHMT1 uncovered no obvious chloroplast or mitochondrial targeting sequences. We functionally expressed AetHMT1 in Escherichia coli and confirmed its biological activity, as evidenced by a positive HMT enzyme activity of 164.516 ± 17.378 nmol min−1 mg−1 protein when catalyzing the transformation of L-homocysteine. Compared with the bacterium containing the empty vector, E. coli harboring the recombinant AetHMT1 plasmid showed much higher tolerance to selenate and selenite. AetHMT1 transcript amounts in different organs were increased by Na2SeO4 treatment, with roots accumulating higher amounts than stems, old leaves and new leaves. We have therefore successfully isolated HMT1 from Ae. tauschii and characterized the biochemical and physiological functions of the corresponding protein.

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Cereal Research Communications
Authors:
Z. L. Li
,
D. D. Wu
,
H. Y. Li
,
G. Chen
,
W. G. Cao
,
S. Z. Ning
,
D. C. Liu
, and
L. Q. Zhang

Gliadin is a main component of gluten proteins that affect functional properties of bread making and contributes to the viscous nature of doughs. In this study, thirteen novel ω-gliadin genes were identified in several Triticum species, which encode the ARH-, ATDand ATN-type proteins. Two novel types of ω-gliadins: ATD- and ATN- have not yet been reported. The lengths of 13 sequences were ranged from 927 to 1269 bp and the deduced mature proteins were varied from 309 to 414 residues. All 13 genes were pseudogenes because of the presence of internal stop codons. The primary structure of these ω-gliadin genes included a signal peptide, a conserved N-terminal domain, a repetitive domain and a conserved C-terminus. In this paper, we first characterize ω-gliadin genes from T. timopheevi ssp. timopheevi and T. timopheevi ssp. araraticum. The ω-gliadin gene variation and the evolutionary relationship of ω-gliadin family genes were also discussed.

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Red coleoptile is an easily observed agronomic trait of wheat and has been extensively studied. However, the molecular mechanism of this trait has not yet been revealed. In this study, the MYB gene TaMYB-D1 was isolated from the wheat cultivar ‘Gy115’, which possesses red coleoptiles. This gene resided at the short arm of the homoelogous group 7 chromosomes. TaMYB-D1 was the only gene expressed in the coleoptiles of ‘Gy115’ and was not expressed in ‘Opata’ and ‘CS’, which have uncoloured coleoptiles. Phylogenetic analysis placed TaMYB-D1 very close to ZmC1 and other MYB proteins regulating anthocyanin biosynthesis. The encoded protein of TaMYB-D1 had an integrated DNA binding domain of 102 amino acids and a transcription domain with 42 amino acids, similar to the structure of ZmC1. Transient expression analysis in onion epidermal cells showed that TaMYB-D1 was located at the plant nucleus, which suggested its role as a transcription factor. The expression of TaMYB-D1 was accompanied with the expression of TaDFR and anthocyanin biosynthesis in the development of the coleoptile of ‘Gy115’. Transient expression analysis showed that only TaMYB-D1 induced a few ‘Opata’ coleoptile cells to synthesize anthocyanin in light, and the gene also induced a colour change to red in many cells with the help of ZmR. All of these results suggested TaMYB-D1 as the candidate gene for the red coleoptile trait of ‘Gy115’.

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To study the development of starch granules in polyploid wheats, we investigated the expression of starch synthetic genes between the synthetic hexaploid wheat SHW-L1, its parents T. turgidum AS2255 and diploid Ae. tauschii AS60. The synthetic hexaploid wheat SHW-L1 showed significantly higher starch content and grain weight than its parents. Scanning electron microscopy (SEM) showed that SHW-L1 rapidly developed starch granules than AS2255 and AS60. The amount of B-type granule in AS60 was less than that in SHW-L1 and AS2255. RT-qPCR result showed that the starch synthetic genes AGPLSU1, AGPLSU2, AGPSSU1, AGPSSU2, GBSSI, SSIII, PHO1 and PHO2 expressed at earlier stages with larger quantity in SHW-L1 than in its parents during wheat grain development. The expression of the above mentioned genes in AS60 was slower than in SHW-L1 and AS2255. The expression pattern of starch synthase genes was also associated with the grain weight and starch content in all three genotypes. The results suggested that the synthetic hexaploid wheat inherited the pattern of starch granule development and starch synthase gene expression from tetraploid parent. The results suggest that tetraploid wheat could plays more important role for starch quality improvement in hexaploid wheat.

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