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1. A mitochondrial phosphate transporter, McPht gene, confers an acclimation regulation of the transgenic rice to phosphorus deficiency
HAN Jiao, YU Guo-hong, WANG Li, LI Wei, HE Rui, WANG Bing, HUANG Sheng-cai, CHENG Xian-guo
Journal of Integrative Agriculture    2018, 17 (09): 1932-1945.   DOI: 10.1016/S2095-3119(17)61792-1
摘要382)      PDF    收藏
Phosphate transporters play an important role in promoting the uptake and transport of phosphate in plants.  In this study, the McPht gene from the Mesembryanthemum crystallinum, a mitochondrial phosphate transporter, was isolated and constructed onto a constitutive expression vector carrying 35S::GFP, and the recombinant constructs were transferred into Oryza sativa japonica L. cv. Kitaake to investigate the regulatory role of the McPht gene under phosphorus deficiency.  The McPht gene encodes a protein of 357 amino acids with six transmembrane domains and is located to the mitochondria, and the mRNA transcripts of the McPht gene are highly accumulated in the shoots of M. crystallinum in response to phosphorus deficiency.  However, more mRNA transcripts of the McPht gene were accumulated in the roots of the transgenic rice under phosphorus deficiency.  Measurements showed that the transgenic rice demonstrated an enhanced promotion in the root development, the root activities, and phosphate uptake under phosphorus deficiency.  Transcriptome sequencing showed that the transgenic rice exhibited total of 198 differentially expressed genes.  Of these, total of 154 differentially expressed genes were up-regulated and total 44 genes were down-regulated comparing to the wild type in response to phosphorus deficiency.  The selective six genes of the up-regulated differentially expressed genes showed an enhanced increase in mRNA transcripts in response to phosphorus deficiency, however, the transcripts of the mitochondrial carrier protein transporter in rice, a homologous gene of the McPht,  in both the transgenic line and the wild type had no obvious differences.  Functional enrichment analyses revealed that the most of the up-regulated genes are involved in the cytoplasmic membrane-bounded vesicle, and most of the down-regulated genes are involved in the mitochondrion and cytoplasmic membrane-bounded vesicle.  The differentially expressed genes were highly enriched in plant secondary metabolisms and plant-pathogen interaction.  These results indicated that the overexpression of the McPht gene might participate in the physiological adaptive modulation of the transgenic rice to phosphorus deficiency by up- or down-regulating the differentially expressed genes.
 
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2. Thellungiella halophila ThPIP1 gene enhances the tolerance of the transgenic rice to salt stress
QIANG Xiao-jing, YU Guo-hong, JIANG Lin-lin, SUN Lin-lin, ZHANG Shu-hui, LI Wei, CHENG Xian-guo
Journal of Integrative Agriculture    2015, 14 (10): 1911-1922.   DOI: 10.1016/S2095-3119(15)61045-0
摘要1773)      PDF    收藏
Aquaporin proteins were demonstrated to play an important regulatory role in transporting water and other small molecules. To better understand physiological functions of aquaporins in extremophile plants, a novel ThPIP1 gene from the Thellungiella halophila was isolated and functionally characterized in the transgenic rice. Data showed that the ThPIP1 protein encoded 284 amino acids, and was identified to be located on the plasma membrane. The expression of ThPIP1 gene in the shoots and roots of T. halophila seedlings were induced by high salinity. The transgenic rice overexpressing ThPIP1 gene significantly increased plants tolerance to salt stress through the pathway regulating the osmotic potentials, accumulation of organic small molecules substances and the ratio of K+/Na+ in the plant cells. Moreover, split-ubiquitin yeast two-hybrid assay showed that ThPIP1 protein specifically interacted with ThPIP2 and a non-specific lipid-transfer protein 2, suggesting that ThPIP1 probably play a key role in responding to the reactions of multiple external stimulus and in participating in different physiological processes of plants exposed to salt stress.
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3. Specific Expression of a Novel Nodulin GmN479 Gene in the Infected Cells of Soybean (Glycine max) Nodules
CHENG Xian-guo, WANG Li, WANG He, YU Guo-hong, BAI You-lu , LIU Meng-meng
Journal of Integrative Agriculture    2011, 10 (10): 1512-1524.   DOI: 10.1016/S1671-2927(11)60146-6
摘要1772)      PDF    收藏
A novel nodulin gene, GmN479 genomic clone composing of 3 630 nucleotides was isolated from mature soybean nodules using GmN479 cDNA as a probe by subtractive hybridization procedure. GmN479 encodes 170 amino acids with 2.09 kb nucleotides promoter region, and contains two important upstream promoter elements, one is a conserved cis-acting sequence motif 5´-AAAGAT-3´ controlling nodulin gene expression, and the other is typical CAAT boxes. GmN479 gene has a single zinc-finger C2H2 domain YSCAFCQRGFSNAQALLGGHMNIH and a conserved motif, QALGGHMN in the zinc-finger with a short leucine repeat in the LDLELRLGL motif closed to C-terminal. These two conserved motifs share respectively higher identity with those in the floral regulator SUPERMAN gene, indicating that GmN479 may function as a transcriptional regulator, and is a likely candidate for playing a role in nodule-morphogenesis. Blotting data showed that GmN479 is a single copy presenting in the genome of soybean nodule, and its expression profile is similar to that of Lba, but it is different from that of ENOD2. GUS staining showed that GmN479 promoter just functions in the infected cells of nodules, indicating that the GmN479 is one of the truly symbiotically induced host genes, and belongs to a late nodulin gene. The expression pattern of GmN479 gene seems to imply that it may be closely related to the development of the nodule. In a sense, it may be a useful marker for identifying the development of the infected cell system in the nodules of soybean.
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