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1.
Ming-Tsair Chan Hsin-Hsiung Chang Shin-Lon Ho Wu-Fu Tong Su-May Yu 《Plant molecular biology》1993,22(3):491-506
We have successfully transferred and expressed a reporter gene driven by an -amylase promoter in a japonica type of rice (Oryza sativa L. cv. Tainung 62) using the Agrobacterium-mediated gene transfer system. Immature rice embryos (10–12 days after anthesis) were infected with an Agrobacterium strain carrying a plasmid containing chimeric genes of -glucuronidase (uidA) and neomycin phosphotransferase (nptII). Co-incubation of potato suspension culture (PSC) with the Agrobacterium inoculum significantly improved the transformation efficiency of rice. The uidA and nptII genes, which are under the control of promoters of a rice -amylase gene (Amy8) and Agrobacterium nopaline synthase gene (nos), respectively, were both expressed in G418-resistant calli and transgenic plants. Integration of foreign genes into the genomes of transgenic plants was confirmed by Southern blot analysis. Histochemical localization of GUS activity in one transgenic plant (R0) revealed that the rice -amylase promoter functions in all cell types of the mature leaves, stems, sheaths and roots, but not in the very young leaves. This transgenic plant grew more slowly and produced less seeds than the wild-type plant, but its R1 and R2 progenies grew normally and produced as much seeds as the wild-type plant. Inheritance of foreign genes to the progenies was also confirmed by Southern blot analysis. These data demonstrate successful gene transfer and sexual inheritance of the chimeric genes. 相似文献
2.
Shin-Lon Ho Li-Fen Huang Chung-An Lu Siou-Luan He Chun-Chin Wang Sheng-Ping Yu Jychian Chen Su-May Yu 《Plant molecular biology》2013,81(4-5):347-361
Germination followed by seedling growth constitutes two essential steps in the initiation of a new life cycle in plants, and in cereals, completion of these steps is regulated by sugar starvation and the hormone gibberellin. A calcium-dependent protein kinase 1 gene (OsCDPK1) was identified by differential screening of a cDNA library derived from sucrose-starved rice suspension cells. The expression of OsCDPK1 was found to be specifically activated by sucrose starvation among several stress conditions tested as well as activated transiently during post-germination seedling growth. In gain- and loss-of-function studies performed with transgenic rice overexpressing a constitutively active or RNA interference gene knockdown construct, respectively, OsCDPK1 was found to negatively regulate the expression of enzymes essential for GA biosynthesis. In contrast, OsCDPK1 activated the expression of a 14-3-3 protein, GF14c. Overexpression of either constitutively active OsCDPK1 or GF14c enhanced drought tolerance in transgenic rice seedlings. Hence, our studies demonstrated that OsCDPK1 transduces the post-germination Ca2+ signal derived from sugar starvation and GA, refines the endogenous GA concentration and prevents drought stress injury, all essential functions to seedling development at the beginning of the life cycle in rice. 相似文献
3.
4.
Chern CG Fan MJ Yu SM Hour AL Lu PC Lin YC Wei FJ Huang SC Chen S Lai MH Tseng CS Yen HM Jwo WS Wu CC Yang TL Li LS Kuo YC Li SM Li CP Wey CK Trisiriroj A Lee HF Hsing YI 《Plant molecular biology》2007,65(4):427-438
With the completion of the rice genome sequencing project, the next major challenge is the large-scale determination of gene
function. As an important crop and a model organism, rice provides major insights into gene functions important for crop growth
or production. Phenomics with detailed information about tagged populations provides a good tool for functional genomics analysis.
By a T-DNA insertional mutagenesis approach, we have generated a rice mutant population containing 55,000 promoter trap and
gene activation or knockout lines. Approximately 20,000 of these lines have known integration sites. The T0 and T1 plants
were grown in net “houses” for two cropping seasons each year since 2003, with the mutant phenotypes recorded. Detailed data
describing growth and development of these plants, in 11 categories and 65 subcategories, over the entire four-month growing
season are available in a searchable database, along with the genetic segregation information and flanking sequence data.
With the detailed data from more than 20,000 T1 lines and 12 plants per line, we estimated the mutation rates of the T1 population,
as well the frequency of the dominant T0 mutants. The correlations among different mutation phenotypes are also calculated.
Together, the information about mutant lines, their integration sites, and the phenotypes make this collection, the Taiwan
Rice Insertion Mutants (TRIM), a good resource for rice phenomics study. Ten T2 seeds per line can be distributed to researchers
upon request.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Chyr-Guan Chern, Ming-Jen Fan, and Su-May Yu have contributed equally to this work. 相似文献
5.
Promoters play key roles in conferring temporal, spatial, chemical, developmental, or environmental regulation of gene expression.
Promoters that are subject to specific regulations are useful for manipulating foreign gene expression in plant cells, tissues,
or organs with desirable patterns and under controlled conditions, and have been important for both basic research and applications
in agriculture biotechnology. Recent advances in genomics technologies have greatly facilitated identification and study of
promoters in a genome scale with high efficiency. Previously we have generated a large T-DNA tagged rice mutant library (TRIM),
in which the T-DNA was designed with a gene/promoter trap system, by placing a promoter-less GUS gene next to the right border of T-DNA. GUS activity screens of this library offer in situ and in planta identifications
and analyses of promoter activities in their native configurations in the rice genome. In the present study, we systematically
performed GUS activity screens of the rice mutant library for genes/promoters constitutively, differentially, or specifically
active in vegetative and reproductive tissues. More than 8,200 lines have been screened, and 11% and 22% of them displayed
GUS staining in vegetative tissues and in flowers, respectively. Among the vegetative tissue active promoters, the ratio of
leaf active versus root active is about 1.6. Interestingly, all the flower active promoters are anther active, but with varied
activities in different flower tissues. To identify tissue specific ABA/stress up-regulated promoters, we compared microarray
data of ABA/stress induced genes with those of tissue-specific expression determined by promoter trap GUS staining. Following
this approach, we showed that the peroxidase 1 gene promoter was ABA up-regulated by 4 fold within 1 day of exposure to ABA and its expression is lateral root specific.
We suggest that this be an easy bioinformatics approach in identifying tissue/cell type specific promoters that are up-regulated
by hormones or other factors.
Su-May Yu and Swee-Suak Ko equally contributed to this work. 相似文献
6.
Human serum albumin (HSA) is the most widely used clinical serum protein. Currently, commercial HSA can only be obtained from
human plasma, due to lack of commercially feasible recombinant protein expression systems. In this study, inducible expression
and secretion of HSA by transformed rice suspension cell culture was established. Mature form of HSA was expressed under the
control of the sucrose starvation-inducible rice α Amy3 promoter, and secretion of HSA into the culture medium was achieved by using the α Amy3 signal sequence. High concentrations of HSA were secreted into culture medium in a short time (2–4 days) by sucrose depletion
after cell concentrations had reached a peak density in culture medium containing sucrose. The recombinant HSA had the same
electrophoretic mobility as commercial HSA and was stable and free from apparent proteolysis in the culture medium. In a flask
scale culture with repeated sucrose provision-depletion cycles, HSA was stably produced with yields up to 11.5% of total medium
proteins or 15 mg/L per cycle after each sucrose provision-depletion cycle. A bubble column type bioreactor was designed for
production of HSA. In the bioreactor scale culture, HSA was produced with yields up to 76.4 mg/L 4 days after sucrose depletion.
HSA was purified from the culture medium to high purity by a simple purification scheme. Enrichment of HSA in culture medium
simplifies downstream purification, minimizes protease degradation, and may reduce production cost. The combination of a DNA
construct containing the α Amy3 promoter and signal sequence, and the use of a rice suspension cell culture can provide an effective system for the production
of recombinant pharmaceutical proteins. 相似文献
7.
8.
Chung-Shen Wu Wei-Tin Kuo Chia-Yu Chang Jun-Yi Kuo Yi-Ting Tsai Su-May Yu Hsi-Ten Wu Peng-Wen Chen 《Plant molecular biology》2014,85(1-2):147-161
Expression of α-amylase genes in rice is induced not only by sugar starvation and gibberellin (GA) but also by O2 deficiency. Promoters of two rice α-amylase genes, αAmy3 and αAmy8, have been shown to direct high-level production of recombinant proteins in rice suspension cells and germinated seeds. In the present study, we modified the cis-acting DNA elements within the sugar/GA response complex (SRC/GARC) of αAmy8 promoter. We found that addition of a G box and duplicated TA box leads to high-level expression of αAmy8 SRC/GARC and significantly enhances αAmy8 promoter activity in transformed rice cells and germinated transgenic rice seeds. We also show that these modifications have drastically increased the activity of αAmy8 promoter in rice seedlings under hypoxia. Our results reveal that the G box and duplicated TA box may play important roles in stimulating promoter activity in response to hypoxia in rice. The modified αAmy8 promoter was used to produce the recombinant human epidermal growth factor (hEGF) in rice cells and hypoxic seedlings. We found that the bioactive recombinant hEGF are stably produced and yields are up to 1.8 % of total soluble protein (TSP) in transformed rice cells. The expression level of synthetic hEGF containing preferred rice codon usage comprises up to 7.8 % of TSP in hypoxic transgenic seedlings. Our studies reveal that the modified αAmy8 promoter can be applicable in establishing a novel expression system for the high-level production of foreign proteins in transgenic rice cells and seedlings under hypoxia. 相似文献
9.
Su-May Yu Yi-Ching Lee Su-Chiung Fang Ming-Tsair Chan Soon-Far Hwa Li-Fei Liu 《Plant molecular biology》1996,30(6):1277-1289
The molecular mechanisms that initiate and control the metabolic activities of seed germination are largely unknown. Sugars may play important roles in regulating such metabolic activities in addition to providing an essential carbon source for the growth of young seedlings and maintaining turgor pressure for the expansion of tissues during germination. To test this hypothesis, we investigated the physiological role of sugars in the regulation of -amylase gene expression and carbohydrate metabolism in embryo and endosperm of germinating rice seeds. RNA gel blot analysis revealed that in the embryo and aleurone cells, expression of four -amylase genes was differentially regulated by sugars via mechanisms beyond the well-known hormonal control mechanism. In the aleurone cells, expression of these -amylase genes was regulated by gibberellins produced in the embryo and by osmotically active sugars. In the embryo, expression of two -amylase genes and production of gibberellins were transient, and were probably induced by depletion of sugars in the embryo upon imbibition, and suppressed by sugars influx from the endosperm as germination proceeded. The differential expression of the four -amylase genes in the embryo and aleurone cells was probably due to their markedly different sensitivities to changes in tissue sugar levels. Our study supports a model in which sugars regulate the expression of -amylase genes in a tissue-specific manner: via a feedback control mechanism in the embryo and via an osmotic control mechanism in the aleurone cells. An interactive loop among sugars, gibberellins, and -amylase genes in the germinating cereal grain is proposed. 相似文献
10.
DEAD-box proteins comprise a large family and function in RNA metabolism. Although DEAD-box proteins are highly conserved among eukaryotes, little is known about the role of DEAD-box proteins in rice. In this study, we identified a rice DEAD-box protein, OsRH36, and demonstrated that OsRH36 and AtRH36 are functional orthologs. OsRH36 was expressed ubiquitously throughout the plant and the OsRH36-GFP fusion protein was localized to the nucleus and nucleolus. Furthermore, functional complementation tests among three homologous DEAD-box protein genes indicated that OsRH36 can restore segregation distortion in the Arabidopsis atrh36-1 mutant, but cannot complement the lethal phenotype in the yeast dbp8p mutant. Moreover, mutation of osrh36 also led to defective transmission of the osrh36 allele, as shown for the atrh36 mutants of Arabidopsis. Previously, we have shown that AtRH36 is involved at an early stage of rRNA maturation and is required for synchronous development of female gametophytes in Arabidopsis. Therefore, we suggest that OsRH36 is required for rRNA biogenesis and megagametogenesis in rice, consistent with the function of AtRH36 in Arabidopsis. 相似文献