Biomedical subjects

Ping Che

Publications and source records attributed to Ping Che.

9 recordsLinked to original sources

PubMed2006

Gene expression patterns during somatic embryo development and germination in maize Hi II callus cultures.

Gene expression patterns were profiled during somatic embryogenesis in a regeneration-proficient maize hybrid line, Hi II, in an effort to identify genes that might be used as developmental markers or targets to optimize regeneration steps for recovering maize plants from tissue culture. Gene expression profiles were generated from embryogenic calli induced to undergo embryo maturation and germination. Over 1,000 genes in the 12,060 element arrays showed significant time variation during somatic embryo development. A substantial number of genes were downregulated during embryo maturation, largely histone and ribosomal protein genes, which may result from a slowdown in cell proliferation and growth during embryo maturation. The expression of these genes dramatically recovered at germination. Other genes up-regulated during embryo maturation included genes encoding hydrolytic enzymes (nucleases, glucosidases and proteases) and a few storage genes (an alpha-zein and caleosin), which are good candidates for developmental marker genes. Germination is accompanied by the up-regulation of a number of stress response and membrane transporter genes, and, as expected, greening is associated with the up-regulation of many genes encoding photosynthetic and chloroplast components. Thus, some, but not all genes typically associated with zygotic embryogenesis are significantly up or down-regulated during somatic embryogenesis in Hi II maize line regeneration. Although many genes varied in expression throughout somatic embryo development in this study, no statistically significant gene expression changes were detected between total embryogenic callus and callus enriched for transition stage somatic embryos.

DNA, Plant↗

PubMed2006

Gene expression programs during shoot, root, and callus development in Arabidopsis tissue culture.

Shoots can be regenerated from Arabidopsis (Arabidopsis thaliana) root explants in tissue culture through a two-step process requiring preincubation on an auxin-rich callus induction medium. Regenerating tissues can be directed along different developmental pathways leading to the formation of shoots, new roots, or callus by transferring to the appropriate organ induction medium. Using gene-profiling methods, we identified groups of genes that serve as molecular signatures of the different developmental processes, i.e. genes that were specifically up- or down-regulated on one developmental pathway, but not on others. One transcription factor gene that was up-regulated during early shoot development was RAP2.6L (At5g13330), a member of the ERF (ethylene response factor) subfamily B-4 of the ERF/APETALA2 transcription factor gene family. RAP2.6L functions in shoot regeneration because T-DNA knockdown mutations in the gene reduced the efficiency of shoot formation in tissue culture, but not normal embryo or seedling development. RAP2.6L promoter:beta-glucuronidase fusions demonstrated that the up-regulation of the gene during shoot regeneration was, at least in part, transcriptionally controlled. The promoter:beta-glucuronidase fusions also demonstrated that RAP2.6L expression was localized to the shoot and emerging leaves, but expression declined in the leaf lamina as leaves expanded. T-DNA knockdown mutations in RAP2.6L reduced the expression of many genes that are normally up-regulated during shoot development including CUP-SHAPED COTYLEDON2 that is involved in shoot meristem specification. Thus, RAP2.6L appears to be part of a network involved in regulating the expression of many other genes in shoot regeneration.

Arabidopsis↗

PubMed2004

Quantitative trait loci associated with adventitious shoot formation in tissue culture and the program of shoot development in Arabidopsis.

Arabidopsis ecotypes, Columbia (Col) and Landsberg erecta (Ler), differ in their capacity to regenerate shoots in culture, as do many other cultivars and varieties of the same plant species. Recombinant inbred (RI) lines derived from a cross of Col x Ler were scored for shoot regeneration, and the Arabidopsis genome was scanned using composite interval mapping for loci associated with shoot regeneration. Three QTL were identified--a major one on chromosome 5 in which the Col parent contributed the superior allele and two minor QTL on chromosomes 1 and 4 in which the Ler parent contributed the superior alleles. The RI lines were binned into genotypic pools to isolate the effects of the major QTL on chromosome 5 while holding the minor QTL constant. To identify genes with expression levels that are associated with the allelic state of the major QTL on chromosome 5, oligonucleotide array expression patterns for genes in the LLC pool (Ler alleles at the minor QTL and a Col allele at the major QTL) were compared to those in the LLL pool (Ler alleles at all QTL). The genes that were significantly differentially expressed between the two pools included several encoding transcription factors and signaling or transposon-related proteins.

Arabidopsis↗

PubMed2003

Separation and quantitation of short-chain coenzyme A's in biological samples by capillary electrophoresis.

Because of the importance of coenzyme A's (CoA's or CoASH) in many metabolic processes and the biosynthesis of some carbohydrates and lipids, many methods have been developed to separate and determine their levels in various tissues for metabolism studies, including enzymatic assays, paper chromatography, and high-performance liquid chromatography (HPLC). However, inadequate separation of coexisting CoA's in biological samples was often encountered due to the similarity of their structures. In this paper, we demonstrated for the first time the separation and quantitation of 12 different CoA's by using capillary electrophoresis with UV detection at 254 nm. All 12 CoA's (CoASH, HMG CoA, methylmalonyl CoA, succinyl CoA, methylcrotonyl CoA, isobutyryl CoA, oxidized CoA, acetyl CoA, crotonoyl CoA, n-propzoyl CoA, acetoacetyl CoA, malonyl CoA) were completely separated at -30 kV in a 100 mM NaH2PO4 running buffer containing 0.1% beta-cyclodextrin at pH 6.0. The total separation time was less than 30 min. The signal response was linear over 2 orders of magnitudes (from 1 to 100 nmol), and the detection limits were in the picomole range. The effects of pH, buffer concentration, additives, and operation voltages on sensitivity and resolution were also discussed. This technique, described here, is much more sensitive, faster, and simpler than the published HPLC methods and can potentially be used for mechanistic study in biological systems involving CoA metabolism.

Animals↗

PubMed2003

Cytokinins and shoot development.

Cytokinins promote the unfolding of a complex gene expression program in tissue culture that results in the formation of shoots. Much has been learned about cytokinin signaling in the past few years; the challenge now is to understand how known steps in cytokinin signaling interface with the process of shoot development in culture and in planta. Several ways have been found to block or bypass the requirement for cytokinins, and these findings have revealed key control steps in the shoot development process. Plant control is exercised not only in the promotion of shoot development, but also in its restraint under conditions where shoots are not ordinarily formed.

Cells, Cultured↗

PubMed2003

The role of biotin in regulating 3-methylcrotonyl-coenzyme a carboxylase expression in Arabidopsis.

As a catalytic cofactor, biotin has a critical role in the enzymological mechanism of a number of enzymes that are essential in both catabolic and anabolic metabolic processes. In this study we demonstrate that biotin has additional non-catalytic functions in regulating gene expression in plants, which are biotin autotrophic organisms. Biotin controls expression of the biotin-containing enzyme, methylcrotonyl-coenzyme A (CoA) carboxylase by modulating the transcriptional, translational and/or posttranslational regulation of the expression of this enzyme. The bio1 mutant of Arabidopsis, which is blocked in the de novo biosynthesis of biotin, was used to experimentally alter the biotin status of this organism. In response to the bio1-associated depletion of biotin, the normally biotinylated A-subunit of methylcrotonyl-CoA carboxylase (MCCase) accumulates in its inactive apo-form, and both MCCase subunits hyperaccumulate. This hyperaccumulation occurs because the translation of each subunit mRNA is enhanced and/or because the each protein subunit becomes more stable. In addition, biotin affects the accumulation of distinct charge isoforms of MCCase. In contrast, in response to metabolic signals arising from the alteration in the carbon status of the organism, biotin modulates the transcription of the MCCase genes. These experiments reveal that in addition to its catalytic role as an enzyme cofactor, biotin has multiple roles in regulating gene expression.

Arabidopsis↗

PubMed2002

Developmental events and shoot apical meristem gene expression patterns during shoot development in Arabidopsis thaliana.

Critical developmental and gene expression profiles were charted during the formation of shoots from root explants in Arabidopsis tissue culture. Shoot organogenesis is a two-step process involving pre-incubation on an auxin-rich callus induction medium (CIM) during which time root explants acquire competence to form shoots during subsequent incubation on a cytokinin-rich shoot induction medium (SIM). At a histological level, the organization of shoot apical meristems (SAMs) appears to occur during incubation on SIM about the time of shoot commitment, i.e. the transition from hormone-dependent to hormone-independent shoot development. Genes involved in SAM formation, such as SHOOTMERISTEMLESS (STM) and CLAVATA1 (CLV1), were upregulated at about the time of shoot commitment, while WUSCHEL (WUS) was upregulated somewhat earlier. Genes required for STM expression, such as CUP-SHAPED COTYLEDON 1 and 2 (CUC1 and 2) were upregulated prior to shoot commitment. Gene expression patterns were determined for two GFP enhancer trap lines with tissue-specific expression in the SAM, including one line reporting on CUC1 expression. CUC1 was generally expressed in callus tissue during early incubation on SIM, but later CUC1 was expressed more locally in presumptive sites of shoot formation. In contrast, the expression pattern of the enhancer trap lines during zygotic embryogenesis was more localized to the presumptive SAM even in early stages of embryogenesis.

Arabidopsis↗

PubMed2002

Metabolic and environmental regulation of 3-methylcrotonyl-coenzyme A carboxylase expression in Arabidopsis.

3-Methylcrotonyl-coenzyme A carboxylase (MCCase) is a nuclear-encoded, mitochondrial biotin-containing enzyme composed of two types of subunits: the biotinylated MCC-A subunit (encoded by the gene At1g03090) and the non-biotinylated MCC-B subunit (encoded by the gene At4g34030). The major metabolic role of MCCase is in the mitochondrial catabolism of leucine, and it also might function in the catabolism of isoprenoids and the mevalonate shunt. In the work presented herein, the single-copy gene encoding the Arabidopsis MCC-A subunit was isolated and characterized. It contains 15 exons separated by 14 introns. We examined the expression of the single-copy MCC-A and MCC-B genes in Arabidopsis by monitoring the accumulation of the two protein and mRNA products. In addition, the expression of these two genes was studied in transgenic plants containing the 1.1- and 1.0-kb 5' upstream sequences of the two MCCase subunit genes, respectively, fused to the beta-glucuronidase gene. Light deprivation induces MCCase expression, which is suppressed by exogenous carbohydrates, especially sucrose. Several lines of evidence indicate that the suppressor of MCCase expression is synthesized in illuminated photosynthetic organs, and can be translocated to other organs to regulate MCCase expression. These results are consistent with the hypothesis that the suppressor of MCCase expression is a carbohydrate, perhaps sucrose or a carbohydrate metabolite. We conclude that MCCase expression is primarily controlled at the level of gene transcription and regulated by a complex interplay between environmental and metabolic signals. The observed expression patterns may indicate that one of the physiological roles of MCCase is to maintain the carbon status of the organism, possibly via the catabolism of leucine.

Arabidopsis↗

PubMed2002

Global and hormone-induced gene expression changes during shoot development in Arabidopsis.

A global analysis of gene expression events during shoot development in Arabidopsis was conducted using oligonucleotide array analysis. Shoots can be induced in tissue culture by preincubating root explants on an auxin-rich callus induction medium (CIM) and by transferring explants to a cytokinin-rich shoot induction medium (SIM), during which time explants become committed to shoot formation and ultimately form shoots. Oligonucleotide array data obtained during shoot development from approximately 8000 Arabidopsis genes were subjected to principal component analysis, which demonstrated that the major components of variation in gene expression during shoot development can be represented by groups of genes, each group of which is upregulated at only one developmental stage. Two percent to three percent of the approximately 8000 Arabidopsis genes monitored in this study were upregulated by fourfold or more at any one stage during shoot development. When upregulated and downregulated genes were categorized by function, it was observed that numerous hormone response genes were upregulated during preincubation on CIM. Groups of genes involved in signaling and/or transcription were induced at or before the time of shoot commitment, and genes that encode components of the photosynthetic apparatus were upregulated later in development before shoot emergence. Primary hormone response genes, such as Aux/IAA genes, were upregulated during preincubation on auxin-rich CIM, and cytokinin-responsive response regulator genes were upregulated during incubation on cytokinin-rich SIM. The expression of ARABIDOPSIS RESPONSE REGULATOR5, a type-A response regulator gene, was upregulated at the time of shoot commitment, and its expression was localized to sites of presumptive shoot formation. Two "hybrid" His kinases involved in cytokinin responses, CRE1, which encodes a cytokinin receptor, and CKI1, a gene that is capable of conferring cytokinin-independent shoot development, were upregulated during incubation on SIM.

Arabidopsis↗