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1.
Sensors (Basel) ; 22(3)2022 Jan 25.
Article in English | MEDLINE | ID: mdl-35161657

ABSTRACT

The RapidIO standard is a packet-switching interconnection technology similar to the Internet Protocol (IP) conceptually. It realizes the high-speed transmission of RapidIO packets at the transport layer, but this greatly increases the probability of network blocking. Therefore, it is of great significance to optimize the RapidIO routing strategy. For this problem, this paper proposes a Double-Antibody Group Multi-Objective Artificial Immune Algorithm (DAG-MOAIA), which improves the local search and global search ability of the population by adaptive crossover and adaptive mutation of the double-antibody groups, and uses co-competition of multi-antibody groups to increase the diversity of population. Through DAG-MOAIA, an optimal transmission path from the source node to multiple destination nodes can be selected to solve the Quality Of Service (QoS) problem during data transmission and ensure the QoS of the RapidIO network. Simulation results show that DAG-MOAIA could obtain high-quality solutions to select better routing transmission paths, and exhibit better comprehensive performance in all simulated test networks, which plays a certain role in solving the problem of the RapidIO routing strategy.


Subject(s)
Algorithms , Computer Communication Networks , Computer Simulation , Mutation
2.
Plant Physiol ; 186(1): 452-468, 2021 05 27.
Article in English | MEDLINE | ID: mdl-33576789

ABSTRACT

Unilateral incompatibility (UI) manifests as pollen rejection in the pistil, typically when self-incompatible (SI) species are pollinated by self-compatible (SC) relatives. In the Solanaceae, UI occurs when pollen lack resistance to stylar S-RNases, but other, S-RNase-independent mechanisms exist. Pistils of the wild tomato Solanum pennellii LA0716 (SC) lack S-RNase yet reject cultivated tomato (Solanum lycopersicum, SC) pollen. In this cross, UI results from low pollen expression of a farnesyl pyrophosphate synthase gene (FPS2) in S. lycopersicum. Using pollen from fps2-/- loss-of-function mutants in S. pennellii, we identified a pistil factor locus, ui3.1, required for FPS2-based pollen rejection. We mapped ui3.1 to an interval containing 108 genes situated on the IL 3-3 introgression. This region includes a cluster of ornithine decarboxylase (ODC2) genes, with four copies in S. pennellii, versus one in S. lycopersicum. Expression of ODC2 transcript was 1,034-fold higher in S. pennellii than in S. lycopersicum styles. Pistils of odc2-/- knockout mutants in IL 3-3 or S. pennellii fail to reject fps2 pollen and abolish transmission ratio distortion (TRD) associated with FPS2. Pollen of S. lycopersicum express low levels of FPS2 and are compatible on IL 3-3 pistils, but incompatible on IL 12-3 × IL 3-3 hybrids, which express both ODC2 and ui12.1, a locus thought to encode the SI proteins HT-A and HT-B. TRD observed in F2 IL 12-3 × IL 3-3 points to additional ODC2-interacting pollen factors on both chromosomes. Thus, ODC2 genes contribute to S-RNase independent UI and interact genetically with ui12.1 to strengthen pollen rejection.


Subject(s)
Ornithine Decarboxylase/genetics , Pollen/physiology , Ribonucleases/genetics , Solanum/physiology , Genes, Plant , Ornithine Decarboxylase/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Ribonucleases/metabolism , Solanum/enzymology
3.
Plant J ; 100(4): 836-850, 2019 11.
Article in English | MEDLINE | ID: mdl-31323151

ABSTRACT

The wild tomato relative Solanum sitiens is a xerophyte endemic to the Atacama Desert of Chile and a potential source of genes for tolerance to drought, salinity and low-temperature stresses. However, until recently, strong breeding barriers prevented its hybridization and introgression with cultivated tomato, Solanum lycopersicum L. We overcame these barriers using embryo rescue, bridging lines and allopolyploid hybrids, and synthesized a library of introgression lines (ILs) that captures the genome of S. sitiens in the background of cultivated tomato. The IL library consists of 56 overlapping introgressions that together represent about 93% of the S. sitiens genome: 65% in homozygous and 28% in heterozygous (segregating) ILs. The breakpoints of each segment and the gaps in genome coverage were mapped by single nucleotide polymorphism (SNP) genotyping using the SolCAP SNP array. Marker-assisted selection was used to backcross selected introgressions into tomato, to recover a uniform genetic background, to isolate recombinant sub-lines with shorter introgressions and to select homozygous genotypes. Each IL contains a single S. sitiens chromosome segment, defined by markers, in the genetic background of cv. NC 84173, a fresh market inbred line. Large differences were observed between the lines for both qualitative and quantitative morphological traits, suggesting that the ILs contain highly divergent allelic variation. Several loci contributing to unilateral incompatibility or hybrid necrosis were mapped with the lines. This IL population will facilitate studies of the S. sitiens genome and expands the range of genetic variation available for tomato breeding and research.


Subject(s)
Genetic Introgression , Solanum lycopersicum/genetics , Solanum/genetics , Desert Climate , Flowers/physiology , Fruit/physiology , Gene Library , Genome, Plant , Solanum lycopersicum/physiology , Plant Breeding , Ploidies , Polymorphism, Single Nucleotide , Recombination, Genetic , Self-Incompatibility in Flowering Plants/genetics
4.
Opt Lett ; 43(16): 3973-3976, 2018 Aug 15.
Article in English | MEDLINE | ID: mdl-30106930

ABSTRACT

Characteristics of intensity noise of optically pumped vertical-cavity surface-emitting lasers and solid-state lasers, induced by the beating of the main lasing mode and non-lasing side modes and its reduction, have been reported in recent years. The mode beating noise of an external cavity diode laser composed of an electrically pumped edge-emitting laser diode chip is studied experimentally in this Letter. The noises due to the beating of the main mode with the first- to third-order side modes are observed, and multiple sub-peaks in the beating noise are measured. It is assumed that the new phenomena are coming from the enhanced four-wave mixing in the longer-active medium. Intensity noise reduction is also demonstrated by using the second harmonic generation of a beta barium borate crystal as a nonlinear absorber.

5.
Plant J ; 93(3): 417-430, 2018 02.
Article in English | MEDLINE | ID: mdl-29206320

ABSTRACT

Multiple independent and overlapping pollen rejection pathways contribute to unilateral interspecific incompatibility (UI). In crosses between tomato species, pollen rejection usually occurs when the female parent is self-incompatible (SI) and the male parent self-compatible (SC) (the 'SI × SC rule'). Additional, as yet unknown, UI mechanisms are independent of self-incompatibility and contribute to UI between SC species or populations. We identified a major quantitative trait locus on chromosome 10 (ui10.1) which affects pollen-side UI responses in crosses between cultivated tomato, Solanum lycopersicum, and Solanum pennelliiLA0716, both of which are SC and lack S-RNase, the pistil determinant of S-specificity in Solanaceae. Here we show that ui10.1 is a farnesyl pyrophosphate synthase gene (FPS2) expressed in pollen. Expression is about 18-fold higher in pollen of S. pennellii than in S. lycopersicum. Pollen with the hypomorphic S. lycopersicum allele is selectively eliminated on pistils of the F1 hybrid, leading to transmission ratio distortion in the F2 progeny. CRISPR/Cas9-generated knockout mutants (fps2) in S. pennelliiLA0716 are self-sterile due to pollen rejection, but mutant pollen is fully functional on pistils of S. lycopersicum. F2 progeny of S. lycopersicum × S. pennellii (fps2) show reversed transmission ratio distortion due to selective elimination of pollen bearing the knockout allele. Overexpression of FPS2 in S. lycopersicum pollen rescues the pollen elimination phenotype. FPS2-based pollen selectivity does not involve S-RNase and has not been previously linked to UI. Our results point to an entirely new mechanism of interspecific pollen rejection in plants.


Subject(s)
Geranyltranstransferase/genetics , Plant Proteins/genetics , Pollen/genetics , Self-Incompatibility in Flowering Plants/genetics , Solanum lycopersicum/physiology , Chromosome Mapping , Chromosomes, Plant , Flowers/genetics , Gene Expression Regulation, Plant , Gene Knockout Techniques , Geranyltranstransferase/metabolism , Loss of Function Mutation , Solanum lycopersicum/genetics , Plant Proteins/metabolism , Plants, Genetically Modified , Ribonucleases/genetics , Ribonucleases/metabolism
6.
Sensors (Basel) ; 17(12)2017 Nov 29.
Article in English | MEDLINE | ID: mdl-29186039

ABSTRACT

Since the Persistent Scatterer Synthetic Aperture Radar (SAR) Interferometry (PSI) technology allows the detection of ground subsidence with millimeter accuracy, it is becoming one of the most powerful and economical means for health diagnosis of major transportation infrastructures. However, structures of different types may suffer from various levels of localized subsidence due to the different structural characteristics and subsidence mechanisms. Moreover, in the complex urban scenery, some segments of these infrastructures may be sheltered by surrounding buildings in SAR images, obscuring the desirable signals. Therefore, the subsidence characteristics on different types of structures should be discussed separately and the accuracy of persistent scatterers (PSs) should be optimized. In this study, the PSI-based subsidence mapping over the entire transportation network of Shanghai (more than 10,000 km) is illustrated, achieving the city-wide monitoring specifically along the elevated roads, ground highways and underground subways. The precise geolocation and structural characteristics of infrastructures were combined to effectively guide more accurate identification and separation of PSs along the structures. The experimental results from two neighboring TerraSAR-X stacks from 2013 to 2016 were integrated by joint estimating the measurements in the overlapping area, performing large-scale subsidence mapping and were validated by leveling data, showing highly consistent in terms of subsidence velocities and time-series displacements. Spatial-temporal subsidence patterns on each type of infrastructures are strongly dependent on the operational durations and structural characteristics, as well as the variation of the foundation soil layers.

7.
Am J Bot ; 103(10): 1847-1861, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27793860

ABSTRACT

PREMISE OF THE STUDY: Self-incompatibility (SI) is a mechanism that prevents inbreeding in many plant species. The mutational breakdown of SI occurs frequently, yet relatively little is known about the evolutionary steps involved in the progressive loss of pistil and pollen SI function. METHODS: In Solanaceae, SI is the S-RNase-based gametophytic type. We used SI and SC populations of the wild tomato species Solanum habrochaites to study natural variation for two pollen SI factors: a Cullin1 (CUL1) protein and an S-locus F-box protein (SLF-23). Pollen compatibility was assessed on an allotriploid tester line encoding an S-RNase recognized by SLF-23. Both pollen factors are required for compatibility on this tester line. Complementation tests and gene sequencing were used to identify mutations in CUL1 or SLF-23. KEY RESULTS: We detected loss-of-function mutations in CUL1 and/or SLF-23 in SC populations collected near the northern and southern geographic margins of this taxon's natural range. Nonmarginal SC and all SI accessions expressed mostly functional alleles of these pollen factors. Comparison of the CUL1 sequences identified several shared deletion mutations present in both northern and southern margin SC accessions. CONCLUSIONS: Loss-of-function mutations in CUL1 and SLF-23 likely became fixed relatively late during SI to SC transitions, after loss of pistil SI function. Mutations in CUL1 establish unilateral incompatibility with SI populations and strengthen reproductive isolation. Point mutations common to northern and southern SC biotypes likely derive from shared ancestral variants found in more central SI populations.


Subject(s)
Cullin Proteins/genetics , Plant Proteins/genetics , Reproductive Isolation , Self-Incompatibility in Flowering Plants , Solanum/physiology , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Cullin Proteins/chemistry , Cullin Proteins/metabolism , DNA, Complementary/genetics , DNA, Complementary/metabolism , Mutation , Phylogeny , Plant Dispersal , Plant Proteins/chemistry , Plant Proteins/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Alignment , Solanum/genetics
8.
BMC Plant Biol ; 16(1): 155, 2016 07 12.
Article in English | MEDLINE | ID: mdl-27405473

ABSTRACT

BACKGROUND: ß-carotene, the most active provitamin A molecule produced by plants, plays important roles in human nutrition and health. ß-carotene does not usually accumulate in the endosperm (i.e. flour) of mature wheat grains, which is a major food source of calories for humans. Therefore, enriching ß-carotene accumulation in wheat grain endosperm will enable a sustainable dietary supplementation of provitamin A. Several metabolic genes affecting ß-carotene accumulation have already been isolated from wheat, including phytoene synthase 1 (PSY1), lycopene ε-cyclase (LCYe) and carotenoid ß-ring hydroxylase1/2 (HYD1/2). RESULTS: In this work, we cloned and biochemically characterized two carotenoid cleavage dioxygenases (CCDs), CCD1 and CCD4, from wheat. While CCD1 homoeologs cleaved ß-apo-8'-carotenal, ß-carotene, lutein and zeaxanthin into apocarotenoid products, CCD4 homoeologs were inactive towards these substrates in in vitro assays. When analyzed by real-time qPCR, PSY1, LCYe, HYD1/2 and CCD1/4 homoeologs showed distinct expression patterns in vegetative tissues and sections of developing tetraploid and hexaploid wheat grains, suggesting that carotenoid metabolic genes and homoeologs are differentially regulated at the transcriptional level in wheat. CONCLUSIONS: The CCD1/4 enzyme activity and the spatial-temporal gene expression data provide critical insights into the specific carotenoid metabolic gene homoeologs that control ß-carotene accumulation in wheat grain endosperm, thus establishing the knowledge base for generation of wheat varieties with enhanced ß-carotene in the endosperm through breeding and genome editing approaches.


Subject(s)
Carotenoids/metabolism , Plant Proteins/metabolism , Seeds/growth & development , Triticum/metabolism , Dioxygenases/genetics , Dioxygenases/metabolism , Gene Expression Regulation, Plant , Geranylgeranyl-Diphosphate Geranylgeranyltransferase/genetics , Geranylgeranyl-Diphosphate Geranylgeranyltransferase/metabolism , Intramolecular Lyases/genetics , Intramolecular Lyases/metabolism , Plant Proteins/genetics , Seeds/enzymology , Seeds/genetics , Seeds/metabolism , Triticum/enzymology , Triticum/genetics , Triticum/growth & development
9.
PLoS One ; 11(5): e0156319, 2016.
Article in English | MEDLINE | ID: mdl-27227328

ABSTRACT

Hydrolyzable tannins (HTs) play important roles in plant herbivore deterrence and promotion of human health. A critical step in HT production is the formation of 1-O-galloyl-ß-D-glucopyranoside (ß-glucogallin, ester-linked gallic acid and glucose) by a UDP-glucosyltransferase (UGT) activity. We cloned and biochemically characterized four candidate UGTs from pomegranate (Punica granatum), of which only UGT84A23 and UGT84A24 exhibited ß-glucogallin forming activities in enzyme assays. Although overexpression and single RNAi knockdown pomegranate hairy root lines of UGT84A23 or UGT84A24 did not lead to obvious alterations in punicalagin (the prevalent HT in pomegranate) accumulation, double knockdown lines of the two UGTs resulted in largely reduced levels of punicalagins and bis-hexahydroxydiphenyl glucose isomers. An unexpected accumulation of galloyl glucosides (ether-linked gallic acid and glucose) was also detected in the double knockdown lines, suggesting that gallic acid was utilized by an unidentified UGT activity for glucoside formation. Transient expression in Nicotiana benthamiana leaves and immunogold labeling in roots of pomegranate seedlings collectively indicated cytosolic localization of UGT84A23 and UGT84A24. Overall, functional characterization and localization of UGT84A23 and UGT84A24 open up opportunities for further understanding the regulatory control of HT metabolism in plants and its coordination with other biochemical pathways in the metabolic network.


Subject(s)
Gallic Acid/metabolism , Glycosyltransferases/metabolism , Hydrolyzable Tannins/metabolism , Lythraceae/metabolism , Plants, Genetically Modified/metabolism , Glycosyltransferases/genetics , Lythraceae/genetics , Lythraceae/growth & development , Phylogeny , Plant Extracts/metabolism , Plant Roots/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/growth & development
10.
Biomed Mater ; 9(1): 015014, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24457395

ABSTRACT

As one of the most important potential candidate alloys for vascular stent application, Mg-Y-Zr based Mg-4.2wt%Y-2.4wt%Nd-0.6wt%Ce(La)-0.5wt%Zr (WE43) alloys were investigated in combination with the forming processes of micro-tubes with 2.0 mm diameter and 0.1 mm wall thickness. Orthogonal experimental design for alloy composition, vacuum melting ingot, heat treatment, integrated plastic deformation and micro-tube forward extrusion are included in the processing procedures. Significant improvements in both the mechanical properties and corrosion resistance in phosphate buffered saline solution for WE43 alloys were achieved through this processing sequence. The influence of the heat treatment and hot extrusion on in vitro degradation and plasticity was found to be associated with grain size reduction and the redistribution of intermetallic particles within the microstructure. As a result, the mechanical properties and the corrosion resistance of Mg alloys can be improved through fine-grain strengthening and solid-solution strengthening to some extent.


Subject(s)
Biocompatible Materials/chemistry , Stents , Alloys/chemistry , Corrosion , Hot Temperature , Magnesium/chemistry , Materials Testing , Metals/chemistry , Pressure , Stress, Mechanical , Tensile Strength , Vascular Diseases/therapy , Yttrium/chemistry , Zirconium/chemistry
11.
Plant Mol Biol ; 80(6): 631-46, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23015203

ABSTRACT

Carotenoid ß-hydroxylases attach hydroxyl groups to the ß-ionone rings (ß-rings) of carotenoid substrates, resulting in modified structures and functions of carotenoid molecules. We cloned and characterized two genes (each with three homeologs), HYD1 and HYD2, which encode ß-hydroxylases in wheat. The results from bioinformatic and nested degenerate PCR analyses collectively suggest that HYD1 and HYD2 may represent the entire complement of non-heme di-iron ß-hydroxylases in wheat. The homeologs of wheat HYDs exhibited major ß-ring and minor ε-ring hydroxylation activities in carotenoid-accumulating E. coli strains. Distinct expression patterns were observed for different HYD genes and homeologs in vegetative tissues and developing grains of tetraploid and hexaploid wheat, suggesting their functional divergence and differential regulatory control in tissue-, grain development-, and ploidy-specific manners. An intriguing observation was that the expression of HYD1, particularly HYD-B1, reached highest levels at the last stage of tetraploid and hexaploid grain development, suggesting that carotenoids (at least xanthophylls) were still actively synthesized in mature grains. This result challenges the common perception that carotenoids are simply being turned over during wheat grain development after their initial biosynthesis at the early grain development stages. Overall, this improved understanding of carotenoid biosynthetic gene expression and carotenoid metabolism in wheat grains will contribute to the improvement of the nutritional value of wheat grains for human consumption.


Subject(s)
Carotenoids/metabolism , Genes, Plant , Mixed Function Oxygenases/genetics , Mixed Function Oxygenases/metabolism , Triticum/enzymology , Triticum/genetics , Cloning, Molecular , Gene Expression Regulation, Developmental , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Humans , Nutritive Value/genetics , Phylogeny , Polyploidy , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Species Specificity , Triticum/growth & development
12.
Planta ; 234(4): 737-48, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21626149

ABSTRACT

Carotenoids perform many critical functions in plants, animals, and humans. It is therefore important to understand carotenoid biosynthesis and its regulation in plants. Phytoene synthase (PSY) catalyzes the first committed and rate-limiting step in carotenoid biosynthesis. While PSY is present as a single copy gene in Arabidopsis, duplicated PSY genes have been identified in many economically important monocot and dicot crops. CmPSY1 was previously identified from melon (Cucumis melo L.), but was not functionally characterized. We isolated a second PSY gene, CmPSY2, from melon in this work. CmPSY2 possesses a unique intron/exon structure that has not been observed in other plant PSYs. Both CmPSY1 and CmPSY2 are functional in vitro, but exhibit distinct expression patterns in different melon tissues and during fruit development, suggesting differential regulation of the duplicated melon PSY genes. In vitro chloroplast import assays verified the plastidic localization of CmPSY1 and CmPSY2 despite the lack of an obvious plastid target peptide in CmPSY2. Promoter motif analysis of the duplicated melon and tomato PSY genes and the Arabidopsis PSY revealed distinctive cis-regulatory structures of melon PSYs and identified gibberellin-responsive motifs in all PSYs except for SlPSY1, which has not been reported previously. Overall, these data provide new insights into the evolutionary history of plant PSY genes and the regulation of PSY expression by developmental and environmental signals that may involve different regulatory networks.


Subject(s)
Chloroplasts/enzymology , Cucumis melo/enzymology , Gene Expression Regulation, Plant/genetics , Geranylgeranyl-Diphosphate Geranylgeranyltransferase/metabolism , Plant Proteins/metabolism , Amino Acid Sequence , Base Sequence , Biological Evolution , Carotenoids/metabolism , Chloroplasts/metabolism , Cloning, Molecular , Consensus Sequence , Cucumis melo/chemistry , Cucumis melo/genetics , Cucumis melo/metabolism , Exons , Flowers/chemistry , Flowers/enzymology , Flowers/genetics , Flowers/metabolism , Fruit/chemistry , Fruit/enzymology , Fruit/genetics , Fruit/metabolism , Genes, Duplicate , Geranylgeranyl-Diphosphate Geranylgeranyltransferase/chemistry , Geranylgeranyl-Diphosphate Geranylgeranyltransferase/genetics , Introns , Models, Molecular , Molecular Sequence Data , Nucleotide Motifs , Organ Specificity , Phylogeny , Plant Leaves/chemistry , Plant Leaves/enzymology , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Proteins/chemistry , Plant Proteins/genetics , Plant Roots/chemistry , Plant Roots/enzymology , Plant Roots/genetics , Plant Roots/metabolism , Plant Stems/chemistry , Plant Stems/enzymology , Plant Stems/genetics , Plant Stems/metabolism , Promoter Regions, Genetic/genetics , Sequence Alignment , Sequence Analysis, DNA
13.
J Agric Food Chem ; 59(6): 2393-402, 2011 Mar 23.
Article in English | MEDLINE | ID: mdl-21361290

ABSTRACT

Skin burning is a new type of skin damage related to exposure to high pH values during the brushing-waxing postharvest operations that has been observed recently on some newly released peach and nectarine [Prunus persica (L.) Batsch] cultivars. In this work, we described this skin disorder for the first time and studied its triggers and biological basis. Different skin burning susceptibility was observed after screening 21 peach and nectarine cultivars. The stability of the skin phenolic extracts to pH in the range 7-10 was studied by UV-visible spectroscopy. This study demonstrated that fruit skin phenolics are not stable at high pH and that the transformations occurring at high pH are reversible and time-dependent. The changes on the UV-visible absorption spectra at different pH values pointed out the copigmentation of anthocyanins as the mechanism beyond the skin burning disorder. Finally, some recommendations to minimize this postharvest damage are also discussed.


Subject(s)
Anthocyanins/chemistry , Fruit/chemistry , Phenols/chemistry , Plant Diseases/etiology , Prunus/chemistry , Fruit/physiology , Hydrogen-Ion Concentration , Pigmentation , Prunus/physiology
14.
Plant Physiol ; 147(2): 816-22, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18441226

ABSTRACT

Abscisic acid (ABA) is a plant hormone found in all higher plants; it plays an important role in seed dormancy, embryo development, and adaptation to environmental stresses, most notably drought. The regulatory step in ABA synthesis is the cleavage reaction of a 9-cis-epoxy-carotenoid catalyzed by the 9-cis-epoxy-carotenoid dioxygenases (NCEDs). The parasitic angiosperm Cuscuta reflexa lacks neoxanthin, one of the common precursors of ABA in all higher plants. Thus, is C. reflexa capable of synthesizing ABA, or does it acquire ABA from its host plants? Stem tips of C. reflexa were cultured in vitro and found to accumulate ABA in the absence of host plants. This demonstrates that this parasitic plant is capable of synthesizing ABA. Dehydration of detached stem tips caused a big rise in ABA content. During dehydration, 18O was incorporated into ABA from 18O2, indicating that ABA was synthesized de novo in C. reflexa. Two NCED genes, CrNCED1 and CrNCED2, were cloned from C. reflexa. Expression of CrNCEDs was up-regulated significantly by dehydration. In vitro enzyme assays with recombinant CrNCED1 protein showed that the protein is able to cleave both 9-cis-violaxanthin and 9'-cis-neoxanthin to give xanthoxin. Thus, despite the absence of neoxanthin in C. reflexa, the biochemical activity of CrNCED1 is similar to that of NCEDs from other higher plants. These results provide evidence for conservation of the ABA biosynthesis pathway among members of the plant kingdom.


Subject(s)
Abscisic Acid/metabolism , Cuscuta/metabolism , Xanthophylls/metabolism , Base Sequence , Blotting, Northern , Cuscuta/genetics , DNA Primers , DNA, Plant , Genes, Plant , Molecular Sequence Data , Plant Shoots/metabolism
15.
J Biol Chem ; 279(45): 46940-5, 2004 Nov 05.
Article in English | MEDLINE | ID: mdl-15342640

ABSTRACT

Enzymes that are able to oxidatively cleave carotenoids at specific positions have been identified in animals and plants. The first such enzyme to be identified was a nine-cis-epoxy carotenoid dioxygenase from maize, which catalyzes the rate-limiting step of abscisic acid biosynthesis. Similar enzymes are necessary for the synthesis of vitamin A in animals and other carotenoid-derived molecules in plants. In the model plant, Arabidopsis, there are nine hypothetical proteins that share some degree of sequence similarity to the nine-cis-epoxy carotenoid dioxygenases. Five of these proteins appear to be involved in abscisic acid biosynthesis. The remaining four proteins are expected to catalyze other carotenoid cleavage reactions and have been named carotenoid cleavage dioxygenases (CCDs). The hypothetical proteins, AtCCD7 and AtCCD8, are the most disparate members of this protein family in Arabidopsis. The max3 and max4 mutants in Arabidopsis result from lesions in AtCCD7 and AtCCD8. Both mutants display a dramatic increase in lateral branching and are believed to be impaired in the synthesis of an unidentified compound that inhibits axillary meristem development. To determine the biochemical function of AtCCD7, the protein was expressed in carotenoid-accumulating strains of Escherichia coli. The activity of AtCCD7 was also tested in vitro with several of the most common plant carotenoids. It was shown that the recombinant AtCCD7 protein catalyzes a specific 9-10 cleavage of beta-carotene to produce the 10 black triangle down-apo-beta-carotenal (C27) and beta-ionone (C13). When AtCCD7 and AtCCD8 were co-expressed in a beta-carotene-producing strain of E. coli, the 13-apo-beta-carotenone (C18) was produced. The C18 product appears to result from a secondary cleavage of the AtCCD7-derived C27 product. The sequential cleavages of beta-carotene by AtCCD7 and AtCCD8 are likely the initial steps in the synthesis of a carotenoid-derived signaling molecule that is necessary for the regulation lateral branching.


Subject(s)
Arabidopsis Proteins/chemistry , Carotenoids/chemistry , Dioxygenases/chemistry , Oxygen/chemistry , Oxygenases/chemistry , Arabidopsis , Catalysis , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Cloning, Molecular , Escherichia coli/metabolism , Kinetics , Mass Spectrometry , Models, Chemical , Mutation , Recombinant Proteins/chemistry , Signal Transduction , Time Factors , beta Carotene/chemistry
17.
Plant Physiol ; 128(2): 544-51, 2002 Feb.
Article in English | MEDLINE | ID: mdl-11842158

ABSTRACT

The plant hormone abscisic acid (ABA) plays important roles in seed maturation and dormancy and in adaptation to a variety of environmental stresses. An effort to engineer plants with elevated ABA levels and subsequent stress tolerance is focused on the genetic manipulation of the cleavage reaction. It has been shown in bean (Phaseolus vulgaris) that the gene encoding the cleavage enzyme (PvNCED1) is up-regulated by water stress, preceding accumulation of ABA. Transgenic wild tobacco (Nicotiana plumbaginifolia Viv.) plants were produced that overexpress the PvNCED1 gene either constitutively or in an inducible manner. The constitutive expression of PvNCED1 resulted in an increase in ABA and its catabolite, phaseic acid (PA). When the PvNCED1 gene was driven by the dexamethasone (DEX)-inducible promoter, a transient induction of PvNCED1 message and accumulation of ABA and PA were observed in different lines after application of DEX. Accumulation of ABA started to level off after 6 h, whereas the PA level continued to increase. In the presence of DEX, seeds from homozygous transgenic line TN1 showed a 4-d delay in germination. After spraying with DEX, the detached leaves from line TN1 had a drastic decrease in their water loss relative to control leaves. These plants also showed a marked increase in their tolerance to drought stress. These results indicate that it is possible to manipulate ABA levels in plants by overexpressing the key regulatory gene in ABA biosynthesis and that stress tolerance can be improved by increasing ABA levels.


Subject(s)
Abscisic Acid/metabolism , Nicotiana/enzymology , Oxygenases/genetics , Plant Growth Regulators/metabolism , Water/pharmacology , Adaptation, Physiological/genetics , Dexamethasone/pharmacology , Dioxygenases , Disasters , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Germination/drug effects , Plant Leaves/drug effects , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Proteins , Plants, Genetically Modified , Seeds/enzymology , Seeds/genetics , Nicotiana/drug effects , Nicotiana/genetics , Water/metabolism
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