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Plant Physiol than nontolerant accumulate to than in tolerant ecotypes


Taiz Plant Physiol 109 1995 was not copper tolerance to differences it should be possible to correlate with at shown in Table I The represents the rate between h at the untreated each ecotype a correl

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Document on Subject : "Plant Physiol than nontolerant accumulate to than in tolerant ecotypes"— Transcript:

1 Plant Physiol. than nontolerant accumula
Plant Physiol. than nontolerant accumulate to than in tolerant ecotypes, a higher metal ions the latter are low molecular weight, metal-binding bacteria (Robinson al., 1993). divided into Class I exhibit sequence similarity to first identified certain fungi, Agaricus (Bel- been identified (Kagi, 1991). align well either the type, originally isolated from wheat a second type, originally identified been identified (GenBank accession motifs exclusively, (GenBank accession and have recently been to be encoded by multigene Goldsbrough, per- broad range organisms by a variety trace metals, particularly suggests that are involved copper with the highest affinity (Tomsett al., 1989), exhibit increased sensitivity to the pea (Evans et expressed uniformly was metal that the differential expression genes might reported variation copper tolerance Taiz, 1995). tion between seedling copper which PCs are the primary constituent (Harmens al., 1993), significant correlation basal level among the ecotypes previously exhibit inducible tolerance, Ws NPT levels. root extension assays seedl

2 ings were conducted the results previous
ings were conducted the results previously described root extension plates were video camera a large video screen. A apart was chromatography paper used plates with were recorded at the positions (before after treat- ment) were ineasured individual video were digitized a Macintosh software to enhance resolution. With this system it was root-tip position greater than or growth during interval was then the net Normal distribution values determined described previously, except Taiz (1995). tissue utilized for rinsed, quick fro- zen in liquid nitrogen, used immediately for RNA Taiz, 1995). Total RNA from plant by grinding liquid nitrogen buffer consisting salicylic acid, p-mercaptoethanol, 4 citrate. The slurry was diluted 3.5 mL citrate, 10 5 min. water-saturated phenol the mixture room temperature min. Residual proteins were from the aqueous phase with nucleic acids were precipitated Total RNA was then samples from Total RNA until used were designed based accession No. GenBank acces- P-tubulin primers used were Taiz Plant Physiol. 109, 1995 was not copper tolerance to d

3 ifferences it should be possible to corr
ifferences it should be possible to correlate with at shown in Table I. The represents the rate between h at the untreated each ecotype. a correlation, basal were plotted extension rate the same determined. No either basal against either the HNI or However, a plot mRNA levels versus the rate gave a highly nificant correlation (Fig. 2A). was plotted against root extension determine the levels for ecotypes (Columbia, copper concentrations to P-tubulin, plotted along growth data. from samples that treated with shown in ecotype exhibited a (HNI) at Shahdara was (Fig. 3C), intermediate between In terms copper tolerance the three descending order curves are compared mRNAs, it is exhibit very different uniformly expressed all three ecotypes the presence copper. In ecotypes. Columbia sion, like most sensitive to reached a is less to Columbia, shifted to Induction began reached a the HNI. that the were about copper tolerance dopsis, a correlation kinetics experiments. time course ecotypes (Columbia, shown in Columbia root extension by 64% 0.70 1.20 1.70 MT2 mRNA (relative units

4 ) mRNA expression (relative units) Corre
) mRNA expression (relative units) Correlation between copper the untreated control growth levels, expressed normalized fluorometric ecotype. The bars represent first 4 h copper treatment recovered only slightly first 8 h increase thereafter, but at slower rate. were constant throughout the time course. The initial recovery The second, slower mRNA increase the slight the remainder time course. Ws (Fig. (Fig. 4C) first 4 h copper. In recovered �50% their initial the subsequent rapid phase which lasted In the the termination Metallothioneins and Copper Tolerance Tolerance (PW Figure 3. Copper dose-response curves for three Arabidopsis ecotypes Error bars each metal h coincided a decline The effects Ag+, Cd2+, Zn2+, Ni2+, salicylic acid gene expression by any with the may have caused a increase. In contrast, the metals being the that heat salicylic acid shock might alter the metal sensitivity the seedlings, we determined the the subse- quent growth response to copper. pretreatment with also tested, Taiz, 1995). As shown in nor prior exposure copper gave protection

5 against a Cu2+ in In con- (Fig. 6C). Fo
against a Cu2+ in In con- (Fig. 6C). For the initial to copper was reduced final level inhibition after treatment was copper tolerance to be mRNA levels, a role ecotypic differentiation. PC levels ecotypes were estimated from the between total shown in ecotypes. Treat- caused a ecotypes. Limeport Buckhorn exhibited the smallest statistically signifi- had the GSH levels. It is were the only ecotypes previously been to possess copper, raising the possibility or other play a inducible tolerance (see "Discus- GSH levels to a also reflects slightly higher rate GSH. In percentage conversion to NPT to be similar average conversion rates. be the primary regulating the relative copper toler- the copper-induced were plotted against the growth (as percentage the presence a general as in at a confidence level the correlation 109, 1995 8 16 Hours after 8 16 24 32 40 48 Hours after 8 16 24 40 48 Hours after course for gene expres- Cu*+ for were determined bars are the high end and Limeport at the Arabidopsis expresses seedlirig tis- that under standard to be constitutiveLy ex- met

6 al inducible. Growing the dark did not a
al inducible. Growing the dark did not alter pattern (data shown). In 1994). Occasionally, data not shown). AgNO3 CdSO4 Heat Salicvllc Shock Acid AgN03 CdS04 metals, heat shock, and salicylic gene expression, expressed fluorometric: units, Treatments are heat shock, Error bars ”Materials and Methods.” Metallothioneins and Copper Tolerance treatment with Hours after treatment with has pro- the microevolution metal-tolerant ecotypes begins with a mutation in conferred by enhanced by mutations Ten Bookum, been strongly implicated other organisms (Hamer, genes encoding proteins are a wide variety among the genes involved the microevolution Such major encode tran- or regulatory proteins the expression one or testing this quantified the correlated these values with the seedling copper as reflected Consistent with the hypothesis, a ship was observed between copper levels also correlation was statistically significant. the analysis the variation the seedlings. at the metal inducible unpublished data). dose-response experiments. copper concentrations below the HNI, c

7 onsistent role for setting the HNI. More
onsistent role for setting the HNI. Moreover, levels coin- the onset metal stress copper concentration a proportionate studies, the roots were 4 h whereas Colum- rates remained constant. dara and Columbia. These with a role for copper. It were comparable treatment, with rate being slightly faster the other two ecotypes. Thus, the three to dif- to sustain the long-term exposure to copper. 109, 1995 values are expressed (GSH) equivalents (one glutathione were calculated were quantitated addition to treatments noted. Both total non-BSO-treated values (data three independent 195 (3) 15 110 some protection against metal also cor- the stimulatory yeast (Hottiger the ACEl gene encodes a gene expres- sion. In both yeast Cu2+ and Ag+ are heavy metal ion gene expression feature held factor is mutations that enhance either metal or the ACEl protein would be j j Santa Clara NP-GSH thiol Correlation between copper the untreated control growth the presence sulhydryl equivalent ecotypes. Each Error bars represent metal tolerance is likely evidenced by the stimulatory additional mut

8 ations be among minor genes involved met
ations be among minor genes involved metal tolerance. gene expression might it will parallel those Despite the evidence, based measurements, favoring a role for in plant has proven the past, are not and Hutchinson Moreover, subjecting thiol-rich proteins, had no to play a unreliable estimate the purification proteins from in plant be overcome. A second is that different metal tolerance mechanisms operate at different developmental stages. Schultz In the present Arabidopsis, only seedlings were examined. Further needed to with metal mature plants and Copper Tolerance MT2 expression has thus metal stress tolerance mecha- synthesis is the primary ecotypic differences been equated strated that value can than NPT Taiz, 1995). with the the other ecotypes exhibited is manifested This could either Miiller for in the design also wish to acknowledge the with the ments. Finally, to Dr. attention the the cur- Received April Clearance Center: disulfide in samples. Methods Primary structure copper metallothionein. Environ vegetation. Geol trations in Agric Food Metal tolerance in p

9 lants: acclimation mechanisms. DC Adrian
lants: acclimation mechanisms. DC Adriano, Trace Metals. Lewis Publishers, Knecht JA, against a role for Evans KM, Gatehouse JA, Robinson NJ the pea trace metal accumulation: implications for function. Plant (1978) The plants. Annu plant kingdom ability to bind heavy metals JC, Thurman organic acids in roots Zenk MH metal-binding peptides from glutatione specific y-glutamylcysteine transpeptidase (phytochelatin synthase). Phytochelatin accumulation Rev Biochem Verkleij JAC Increased zinc tolerance to increased the genus Physiological char- overexpressing its transcriptional Kruckeburg AR problems. Univ a copper (1993) The metal tolerance The ecological significance hyperaccumulation: a plant chemical defense. Oecologia Taiz Plant Physiol. cucumber seedlings. copper metallothionein mesh transfer for metal tolerance Quantitative assessment soils by trace metals. metal tolerance in the fission yeast requires an cassette-type vacuolar membrane transporter. metal-binding peptides fission yeast ABC-type vac- uolar membrane protein. tolerance in Schizosaccharomyces Pombe. In

10 Vitro Cell Dev 29: 213-219 Stillman M t
Vitro Cell Dev 29: 213-219 Stillman M thiolate clusters in Phytochelatins. Annu Metal-binding polypeptides Heavy Metal Tolerance Plants, Evolutionary As- Kalff MMA Are phytochelatins involved in ferential metal tolerance or reflect metal-im- higher plants. Reeves, eds, Hutchinson TC against a key role for metallothionein-like protein New Phytol activates transcription metallothionein gene. small genome 9 expressed produces a Plant Physiol99: heavy metal-binding peptides Metal tolerance mechanisms in The 4th Heavy Metals metal tolerance. Aspects Molecular biology plants. Plant Cell Environ MD, Barcelo and cadmium cumulate both metals. their role (1989). The role in the copper tolerance Hamer, eds, Metal Ion heavy metal in higher plants. Shaw, ed, Heavy Metal Tolernnce Wagner G J Subcellular localization tobacco leaves. Functional homologs gal metallothionein genes from Sinsheimer Laboratories, Arabidopsis ecotypes were compared nonprotein thiol inducible in were compared I I I I I I I I a a a a a g 2 a a - a a expression extension data. Root extension