bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytokinins in Tobacco and Wheat Chloroplasts. Occurrence and Changes Due to Light/Dark Treatment
Authors: Benková Eva , Witters Erwin , Van Dongen Walter , Kolar Jan , Motyka Václav , Brzobohatý Bretislav , Van Onckelen Harry A. , Machácková Ivana 
Published: July 2002

Abstract
Although cytokinins (CKs) affect a number of processes connected with chloroplasts, it has never been rigorously proven that chloroplasts contain CKs. We isolated intact chloroplasts from tobacco (Nicotiana tabacum L. cv SR1) and wheat (Triticum aestivum L. cv Ritmo) leaves and determined their CKs by liquid chromatography/tandem mass spectroscopy. Chloroplasts from both species contained a whole spectrum of CKs, including free bases (zeatin and isopentenyladenine), ribosides (zeatin riboside, and isopentenyladenosine), ribotides (isopentenyladenosine-5′-monophosphate, zeatin riboside-5′-monophosphate, and dihydrozeatin riboside-5′-monophosphate), and N-glucosides (zeatin-N 9-glucoside, dihydrozeatin-N 9-glucoside, zeatin-N 7-glucoside, and isopentenyladenine-N-glucosides). In chloroplasts there was a moderately higher relative amount of bases, ribosides, and ribotides than in leaves, and a significantly increased level ofN 9-glucosides of zeatin and dihydrozeatin. Tobacco and wheat chloroplasts were prepared from leaves at the end of either a dark or light period. After a dark period, chloroplasts accumulated more CKs than after a light period. The differences were moderate for free bases and ribosides, but highly significant for glucosides. Tobacco chloroplasts from dark-treated leaves contained zeatin riboside-O-glucoside and dihydrozeatin riboside-O-glucoside, as well as a relatively high CK oxidase activity. These data show that chloroplasts contain a whole spectrum of CKs and the enzymatic activity necessary for their metabolism.
Brassinosteroid-Insensitive Dwarf Mutants of Arabidopsis Accumulate Brassinosteroids
Authors: Noguchi Takahiro , Fujioka Shozo , Choe Sunghwa , Takatsuto Suguru , Yoshida Shigeo , Yuan Heng , Feldmann Kenneth A. , Tax Frans E. 
Published: July 2002

Abstract
Seven dwarf mutants resembling brassinosteroid (BR)-biosynthetic dwarfs were isolated that did not respond significantly to the application of exogenous BRs. Genetic and molecular analyses revealed that these were novel alleles ofBRI1 (Brassinosteroid-Insensitive 1), which encodes a receptor kinase that may act as a receptor for BRs or be involved in downstream signaling. The results of morphological and molecular analyses indicated that these represent a range of alleles from weak to null. The endogenous BRs were examined from 5-week-old plants of a null allele (bri1-4) and two weak alleles (bri1-5 and bri1-6). Previous analysis of endogenous BRs in several BR-biosynthetic dwarf mutants revealed that active BRs are deficient in these mutants. However,bri1-4 plants accumulated very high levels of brassinolide, castasterone, and typhasterol (57-, 128-, and 33-fold higher, respectively, than those of wild-type plants). Weaker alleles (bri1-5 and bri1-6) also accumulated considerable levels of brassinolide, castasterone, and typhasterol, but less than the null allele (bri1-4). The levels of 6-deoxoBRs in bri1 mutants were comparable to that of wild type. The accumulation of biologically active BRs may result from the inability to utilize these active BRs, the inability to regulate BR biosynthesis in bri1 mutants, or both. Therefore,BRI1 is required for the homeostasis of endogenous BR levels.
Early Copper-Induced Leakage of K+ from Arabidopsis Seedlings Is Mediated by Ion Channels and Coupled to Citrate Efflux
Authors: Murphy Angus S. , Eisinger William R. , Shaff Jon E. , Kochian Leon V. , Taiz Lincoln 
Published: July 2002

Abstract
Copper tolerance among Arabidopsis ecotypes is inversely correlated with long-term K+ leakage and positively correlated with short-term K+ leakage (A. Murphy, L. Taiz [1997] New Phytol 136: 211–222). To probe the mechanism of the early phase of K+ efflux, we tested various channel blockers on copper and peroxide-induced K+efflux from seedling roots. The K+ channel blockers tetraethyl ammonium chloride and 4-aminopyridine (4-AP) both inhibited short-term copper-induced K+ efflux. In contrast, peroxide-induced K+ efflux was insensitive to both tetraethyl ammonium chloride and 4-AP. Copper-induced lipid peroxidation exhibited a lag time of 4 h, while peroxide-induced lipid peroxidation began immediately. These results suggest that short-term copper-induced K+ efflux is mediated by channels, while peroxide-induced K+ efflux represents leakage through nonspecific lesions in the lipid bilayer. Tracer studies with 86Rb+ confirmed that copper promotes K+ efflux rather than inhibiting K+uptake. Short-term K+ release is electroneutral, since electrophysiological measurements indicated that copper does not cause membrane depolarization. Short-term K+ efflux was accompanied by citrate release, and copper increased total citrate levels. Since citrate efflux was blocked by 4-AP, K+appears to serve as a counterion during copper-induced citrate efflux. As copper but not aluminum selectively induces citrate production and release, it is proposed that copper may inhibit a cytosolic form of aconitase.
Nucleoside Diphosphate Kinase Required for Coleoptile Elongation in Rice
Authors: Pan Ling , Kawai Maki , Yano Akira , Uchimiya Hirofumi 
Published: July 2002

Abstract
Although several nucleoside diphosphate (NDP) kinase genes have been cloned in plants, little is known about the functional significance of this enzyme during plant growth and development. We introduced a chimeric gene encoding an antisense RNA of NDP kinase under the control of the Arabidopsis heat shock protein HSP81-1 promoter into rice (Oryza sativaL.) plants using the Agrobacterium tumefacienstransformation system. The expression of antisense RNA down-regulated the accumulation of mRNA, resulting in reduced enzyme activity even under the standard growth temperature (25°C) in transgenic plants. Following heat shock treatment (37°C), NDP kinase activities in some transgenic rice plants were more reduced than those grown under 25°C. The comparison of the coleoptile growth under submersion showed that cell elongation process was inhibited in antisense NDP kinase transgenic plants, suggesting that an altered guanine nucleotide level may be responsible for the processes.
The Genepat-2, Which Induces Natural Parthenocarpy, Alters the Gibberellin Content in Unpollinated Tomato Ovaries
Authors: Fos Mariano , Nuez Fernando , Garcı́a-Martı́nez José L. 
Published: July 2002

AbstractWe investigated the role of gibberellins (GAs) in the effect of pat-2, a recessive mutation that induces facultative parthenocarpic fruit development in tomato (Lycopersicon esculentum Mill.) using near-isogenic lines with two different genetic backgrounds. Unpollinated wild-type Madrigal (MA/wt) and Cuarenteno (CU/wt) ovaries degenerated, but GA3 application induced parthenocarpic fruit growth. On the contrary, parthenocarpic growth of MA/pat-2 and CU/pat-2 fruits, which occurs in the absence of pollination and hormone application, was not affected by GA3. Pollinated MA/wt and parthenocarpic MA/pat-2 ovary development was negated by paclobutrazol, and this inhibitory effect was counteracted by GA3. The main GAs of the early-13-hydroxylation pathway (GA1, GA3, GA8, GA19, GA20, GA29, GA44, GA53, and, tentatively, GA81) and two GAs of the non-13-hydroxylation pathway (GA9 and GA34) were identified in MA/wt ovaries by gas chromatography-selected ion monitoring. GAs were quantified in unpollinated ovaries at flower bud, pre-anthesis, and anthesis. In unpollinated MA/pat-2 and CU/pat-2ovaries, the GA20 content was much higher (up to 160 times higher) and the GA19 content was lower than in the corresponding non-parthenocarpic ovaries. The application of an inhibitor of 2-oxoglutarate-dependent dioxygenases suggested that GA20 is not active per se. The pat-2mutation may increase GA 20-oxidase activity in unpollinated ovaries, leading to a higher synthesis of GA20, the precursor of an active GA.
Isolation of Arabidopsis Mutants Lacking Components of Acquired Thermotolerance
Authors: Burke John J. , O'Mahony Patrick J. , Oliver Melvin J. 
Published: July 2002

Abstract
Acquired thermotolerance is a complex physiological phenomenon that enables plants to survive normally lethal temperatures. This study characterizes the temperature sensitivity of Arabidopsis using a chlorophyll accumulation bioassay, describes a procedure for selection of acquired thermotolerance mutants, and provides the physiological characterization of one mutant (AtTS02) isolated by this procedure. Exposure of etiolated Arabidopsis seedlings to 48°C or 50°C for 30 min blocks subsequent chlorophyll accumulation and is eventually lethal. Arabidopsis seedlings can be protected against the effects of a 50°C, 30-min challenge by a 4-h pre-incubation at 38°C. By the use of the milder challenge, 44°C for 30 min, and protective pretreatment, mutants lacking components of the acquired thermotolerance system were isolated. Putative mutants isolated by this procedure exhibited chlorophyll accumulation levels (our measure of acquired thermotolerance) ranging from 10% to 98% of control seedling levels following pre-incubation at 38°C and challenge at 50°C. The induction temperatures for maximum acquired thermotolerance prior to a high temperature challenge were the same in AtTS02 and RLD seedlings, although the absolute level of chlorophyll accumulation was reduced in the mutant. Genetic analysis showed that the loss of acquired thermotolerance in AtTS02 was a recessive trait. The pattern of proteins synthesized at 25°C and 38°C in the RLD and AtTS02 revealed the reduction in the level of a 27-kD heat shock protein in AtTS02. Genetic analysis showed that the reduction of this protein level was correlated with the acquired thermotolerance phenotype.
Expression of a Gene Encoding Mitochondrial Aldehyde Dehydrogenase in Rice Increases under Submerged Conditions
Authors: Nakazono Mikio , Tsuji Hiroyuki , Li Yuhua , Saisho Daisuke , Arimura Shin-ichi , Tsutsumi Nobuhiro , Hirai Atsushi 
Published: July 2002

Abstract
It is known that alcoholic fermentation is important for survival of plants under anaerobic conditions. Acetaldehyde, one of the intermediates of alcoholic fermentation, is not only reduced by alcohol dehydrogenase but also can be oxidized by aldehyde dehydrogenase (ALDH). To determine whether ALDH plays a role in anaerobic metabolism in rice (Oryza sativa L. cv Nipponbare), we characterized a cDNA clone encoding mitochondrial ALDH from rice (Aldh2a). Analysis of sub-cellular localization of ALDH2a protein using green fluorescent protein and an in vitro ALDH assay using protein extracts from Escherichia coli cells that overexpressed ALDH2a indicated that ALDH2a functions in the oxidation of acetaldehyde in mitochondria. A Southern-blot analysis indicated that mitochondrial ALDH is encoded by at least two genes in rice. We found that the Aldh2a mRNA was present at high levels in leaves of dark-grown seedlings, mature leaf sheaths, and panicles. It is interesting that expression of the riceAldh2a gene, unlike the expression of the tobacco (Nicotiana tabacum) Aldh2a gene, was induced in rice seedlings by submergence. Experiments with ruthenium red, which is a blocker of Ca2+ fluxes in rice as well as maize (Zea mays), suggest that the induction of expression of Adh1 and Pdc1 by low oxygen stress is regulated by elevation of the cytosolic Ca2+level. However, the induction of Aldh2a gene expression may not be controlled by the cytosolic Ca2+ level elevation. A possible involvement of ALDH2a in the submergence tolerance of rice is discussed.
Rac-Related GTP-Binding Protein in Elicitor-Induced Reactive Oxygen Generation by Suspension-Cultured Soybean Cells
Authors: Park Jumok , Choi Hyun-Jung , Lee Sumin , Lee Taehoon , Yang Zhenbiao , Lee Youngsook 
Published: July 2002

Abstract
Plant cells produce reactive oxygen species (ROS) in response to many stimuli. However, the mechanism of ROS biosynthesis remains unclear. We have explored the hypothesis that the superoxide burst in plants mechanistically resembles the oxidative burst in neutrophils. First we have confirmed that ROS production, which occurs in suspension-cultured soybean (Glycine max) cells in response to hypo-osmotic shock, is inhibited by diphenylene iodonium, an inhibitor of the flavin-dependent oxidase of neutrophils. Because a Rac family G protein is an essential regulator of this NADPH oxidase, and because many plant homologs of Rac have been cloned, we next examined whether Rac-like proteins might be involved in the oxidative burst in the soybean cells. We identified a Rac-like 21-kD soybean protein that cross-reacts with antibodies to human Rac and garden pea Rop and also binds [γ-35S] GTP, a diagnostic trait of small G proteins. This Rac-related protein translocated from the cytosol to microsomes during the oxidative burst. Moreover, soybean cells transiently transformed with either a dominant negative (RacN17) or a dominant positive (RacV12) form of Rac1 showed the anticipated altered responses to three different stimuli: hypo-osmotic shock, oligo-GalUA, and harpin. In response to these stimuli, cells transformed with RacN17 produced less ROS and cells transformed with RacV12 generated more ROS than control cells. These results strongly suggest that a Rac-related protein participates in the regulation of ROS production in soybean cells, possibly via activation of an enzyme complex similar to the NADPH oxidase of phagocytes in animal systems.
Molecular and Biochemical Characterization of a Cytokinin Oxidase from Maize
Authors: Bilyeu Kristin D. , Cole Jean L. , Laskey James G. , Riekhof Wayne R. , Esparza Thomas J. , Kramer Michelle D. , Morris Roy O. 
Published: July 2002

Abstract
It is generally accepted that cytokinin oxidases, which oxidatively remove cytokinin side chains to produce adenine and the corresponding isopentenyl aldehyde, play a major role in regulating cytokinin levels in planta. Partially purified fractions of cytokinin oxidase from various species have been studied for many years, but have yet to clearly reveal the properties of the enzyme or to define its biological significance. Details of the genomic organization of the recently isolated maize (Zea mays) cytokinin oxidase gene (ckx1) and some of its Arabidopsis homologs are now presented. Expression of an intronless ckx1 inPichia pastoris allowed production of large amounts of recombinant cytokinin oxidase and facilitated detailed kinetic and cofactor analysis and comparison with the native enzyme. The enzyme is a flavoprotein containing covalently bound flavin adenine dinucleotide, but no detectable heavy metals. Expression of the oxidase in maize tissues is described.
Comparison of Gas Exchange and Bioassay Determinations of the Ammonia Compensation Point in Luzula sylvatica (Huds.) Gaud.
Authors: Hill Paul W. , Raven John A. , Loubet Benjamin , Fowler David , Sutton Mark A. 
Published: July 2002

Abstract
Determinations of the NH3 compensation point for the understory plant of semi-natural woodlands Luzula sylvatica (Huds.) Gaud. were carried out by measurements of gas exchange and by calculation from the NH4 +concentration and pH of extracts of the foliar apoplast. Compensation points determined by gas exchange measurements were among the lowest yet reported (0.51–1.10 μg NH3 m−3) and those calculated from apoplast extracts were lower than any yet reported (0.017–0.54 μg NH3 m−3). Those determined by gas exchange were consistently found to be between 2 and 30 times higher than those determined from apoplast extracts. Consideration of possible causes of this discrepancy, which is not confined to this investigation, showed that all likely errors would result in an increase in the discrepancy, or were insufficient to account for observed differences. It is suggested that spatial variability of pH and NH4 + concentration within the foliar apoplast represents the most promising line for further investigation. It is also shown that the foliar apoplast of L. sylvatica is sufficiently buffered to eliminate the need for correction of H+ concentration for dilution during extraction, but that it is necessary to correct the NH4 + concentration of apoplast extracts for dilution.
Molecular Structure of Three Mutations at the Maizesugary1 Locus and Their Allele-Specific Phenotypic Effects
Authors: Dinges Jason R. , Colleoni Christophe , Myers Alan M. , James Martha G. 
Published: July 2002

Abstract
Starch production in all plants examined is altered by mutations of isoamylase-type starch-debranching enzymes (DBE), although how these proteins affect glucan polymer assembly is not understood. Various allelic mutations in the maize (Zea mays) genesugary1 (su1), which codes for an isoamylase-type DBE, condition distinct kernel phenotypes. This study characterized the recessive mutations su1-Ref,su1-R4582::Mu1, and su1-st, regarding their molecular basis, chemical phenotypes, and effects on starch metabolizing enzymes. The su1-Ref allele results in two specific amino acid substitutions without affecting the Su1 mRNA level. The su1-R4582::Mu1 mutation is a null allele that abolishes transcript accumulation. Thesu1-st mutation results from insertion of a novel transposon-like sequence, designated Toad, which causes alternative pre-mRNA splicing. Three su1-st mutant transcripts are produced, one that is nonfunctional and two that code for modified SU1 polypeptides. The su1-st mutation is dominant to the null allele su1-R4582::Mu1,but recessive to su1-Ref, suggestive of complex effects involving quaternary structure of the SU1 enzyme. All threesu1- alleles severely reduce or eliminate isoamylase-type DBE activity, although su1-st kernels accumulate less phytoglycogen and Suc than su1-Ref orsu1-R4582::Mu1 mutants. The chain length distribution of residual amylopectin is significantly altered bysu1-Ref and su1-R4582::Mu1, whereas su1-st has modest effects. These results, together with su1 allele-specific effects on other starch- metabolizing enzymes detected in zymograms, suggest that total DBE catalytic activity is the not the sole determinant ofSu1 function and that specific interactions between SU1 and other components of the starch biosynthetic system are required.
Possible Role of Root Border Cells in Detection and Avoidance of Aluminum Toxicity
Authors: Miyasaka Susan C. , Hawes Martha C. 
Published: July 2002

Abstract
Root border cells are living cells that surround root apices of most plant species and are involved in production of root exudates. We tested predictions of the hypothesis that they participate in detection and avoidance of aluminum (Al) toxicity by comparing responses of two snapbean (Phaseolus vulgaris) cultivars (cv Dade and cv Romano) known to differ in Al resistance at the whole-root level. Root border cells of these cultivars were killed by excess Al in agarose gels or in simple salt solutions. Percent viability of Al-sensitive cv Romano border cells exposed in situ for 96 h to 200 μm total Al in an agarose gel was significantly less than that of cv Dade border cells; similarly, relative viability of harvested cv Romano border cells was significantly less than that of cv Dade cells after 24 h in 25 μm total Al in a simple salt solution. These results indicate that Al-resistance mechanisms that operate at the level of whole roots also operate at the cellular level in border cells. Al induced a thicker mucilage layer around detached border cells of both cultivars. Cultivar Dade border cells produced a thicker mucilage layer in response to 25 μM Al compared with that of cv Romano cells after 8 h of treatment and this phenomenon preceded that of observed cultivar differences in relative cell viability. Release of an Al-binding mucilage by border cells could play a role in protecting root tips from Al-induced cellular damage.
Induction of Vacuolar ATPase and Mitochondrial ATP Synthase by Aluminum in an Aluminum-Resistant Cultivar of Wheat
Authors: Hamilton Christie A. , Good Allen G. , Taylor Gregory J. 
Published: July 2002

Abstract
Two 51-kD aluminum (Al)-induced proteins (RMP51, root membrane proteins of 51 kD) were recently discovered in an aluminum-resistant cultivar of wheat (Triticum aestivum) cv PT741 (Basu et al., 1994a). These proteins segregate with the aluminum resistance phenotype in a segregating population arising from a cross between Al-resistant cv PT741 and Al-sensitive cv Katepwa (Taylor et al., 1997). The proteins have been purified by continuous elution electrophoresis and analyzed by peptide microsequencing. Sequence analysis of the purified peptides revealed that they are homologous to the B subunit of the vacuolar H+-ATPase (V-ATPase) and the α- and β-subunits of the mitochondrial ATP synthase (F1F0-ATPase). To confirm that these ATPases are induced by Al, ATPase activity and transcript levels were analyzed under Al stress. Both V-ATPase and F1F0-ATPase activities were induced by Al and responded in a dose-dependent manner to 0 to 150 μm Al. In contrast, plasma membrane H+-ATPase (P-ATPase) activity decreased to 0.5× control levels, even when plants were exposed to 25 μm Al. Northern analysis showed that the transcript encoding the B subunit of V-ATPase increased by 2.2× in a dose-dependent manner, whereas levels of the transcript encoding the α-subunit of F1F0-ATPase remained constant. The effect of Al on ATPase activity in other cultivars was also examined. The Al-resistant cultivar, cv PT741, was the only cultivar to show induction of V- and F1F0-ATPases. These results suggest that the V-ATPase in cv PT741 is responding specifically to Al stress with the ATP required for its activity supplied by ATP synthase to maintain energy balance within the cell.
Nitrite Reductase Gene Enrichment Improves Assimilation of NO2 in Arabidopsis
Authors: Takahashi Misa , Sasaki Yukari , Ida Shoji , Morikawa Hiromichi 
Published: July 2002

AbstractTransgenic plants of Arabidopsis bearing the spinach (Spinacia oleracea) nitrite reductase (NiR, EC 1.7.7.1) gene that catalyzes the six-electron reduction of nitrite to ammonium in the second step of the nitrate assimilation pathway were produced by use of the cauliflower mosaic virus 35S promoter and nopaline synthase terminator. Integration of the gene was confirmed by a genomic polymerase chain reaction (PCR) and Southern-blot analysis; its expression by a reverse transcriptase-PCR and two-dimensional polyacrylamide gel electrophoresis western-blot analysis; total (spinach + Arabidopsis) NiR mRNA content by a competitive reverse transcriptase-PCR; localization of NiR activity (NiRA) in the chloroplast by fractionation analysis; and NO2 assimilation by analysis of the reduced nitrogen derived from NO2(NO2-RN). Twelve independent transgenic plant lines were characterized in depth. Three positive correlations were found for NiR gene expression; between the total NiR mRNA and total NiR protein contents (r = 0.74), between the total NiR protein and NiRA (r = 0.71), and between NiRA and NO2-RN (r = 0.65). Of these twelve lines, four had significantly higher NiRA than the wild-type control (P &lt; 0.01), and three had significantly higher NO2-RN (P &lt; 0.01). Each of the latter three had one to two copies of spinach NiR cDNA per haploid genome. The NiR flux control coefficient for NO2 assimilation was estimated to be about 0.4. A similar value was obtained for an NiR antisense tobacco (Nicotiana tabacum cv Xanthi XHFD8). The flux control coefficients of nitrate reductase and glutamine synthetase were much smaller than this value. Together, these findings indicate that NiR is a controlling enzyme in NO2assimilation by plants.