Heteroblasty in Arabidopsis thaliana (L.) Heynh

 Heteroblasty in Arabidopsis thaliana was analyzed in a variety of plants with mutations in leaf morphology using a tissue-specific β-glucuronidase gene marker. Some mutants exhibited their mutant phenotypes specifically in foliage leaves. The phenotypes associated with the foliage-leaf-specific mutations were also found to be induced ectopically in cotyledons in the presence of the lec1 mutation. Moreover, the features of an emf1lec1 double mutant showed that cotyledons can be partially converted into carpelloids. When heteroblastic traits were examined in foliage leaves in the presence of certain mutations or natural deviations by histochemical analysis of the expression of the tissue-specific marker gene, it was found that ectopic expression of the developmental program for the first foliage leaves in lec1 cotyledons seemed to affect the heteroblastic features of the first set of foliage leaves, while foliage leaves beyond the third position appeared normal. Similarly, in wild-type plants, discrepancies in heteroblastic features, relative to standard features, of foliage leaves at early positions seemed to be eliminated in foliage leaves at later positions. These results suggest that heteroblasty in foliage leaves might be affected in part by the heteroblastic stage of the preceding foliage leaves but is finally controlled autonomously at each leaf position. Received: 9 July 1999 / Accepted: 17 August 1999     

Genphysiologie quantitativer Merkmale bei Arabidopsis thaliana (L.) Heynh.

Zusammenfassung In biometrischen Untersuchungen zur Genetik quantitativer Merkmale wird bisher weitgehend das Problem der Genregulation übersehen. Chlorophyll b-Defektmutanten der Crucifere Arabidopsis thaliana (L.) Heynh. erlauben eine direkte biochemische und physiologische Prüfung ihrer quantitativen Genwirkungen in der Biosynthesekette der Plastidenpigmente. Auf Grund bestimmter Testkreuzungen zwischen zwei Mutanten und der Normalform sowie eines kombinierten Verfahrens zur phänotypischen Klassifizierung bestehend aus präparativer Dünnschichtchromatographie aller Plastidenpigmente, visueller Blattfarbenbonitur, papierchromatographischem Einzelpflanzentest und quantitativer Pigmentmessung der Linien werden in Übereinstimmung mit anderen Autoren die nachweisbaren Genotypen einer Allelenreihe ch ⁺, ch ¹ und ch ² des ch-Locus zugeordnet. ch ¹/ch ¹ blockiert die Chlorophyll b-Synthese vollständig, ch ²/ch ² nur etwa zu 90% gegenüber dem Wildtyp und in der Heterozygote ch ²/ch ¹ wird offenbar die Blockade durch komplementative Effekte noch stärker aufgehoben (Superdominanz des ch ²- über das ch ¹-Allel). Die in den Spaltungsgenerationen erhöhten Chlorophyll- und Carotinoid-Varianzen aller Genotypen gegenüber den Eltern weisen auf einen evtl. polygenen Hintergrund der Gesamtpigmentbildung hin, dessen physiologischer Wirkungsbereich aber relativ zu dem des Hauptgens ch begrenzt ist. Orientierende In-vitro-Versuche ergeben, daß im plastidenfreien Blattgewebe-Extrakt des Wildtyps ch ⁺/ch ⁺ ein Wirkstoffprinzip vorhanden ist, das belichtete Chloroplasten der in vivo pigmentdefekten Genotypen ch ¹/ch ¹ und ch ²/ch ² zur Chlorophyll b-Synthese befähigt. Weiterführende Aussagen über Art, Umfang und Zeitpunkt der hierbei angenommenen Enzymaktivitäts- bzw. Substratsänderungen werden von Versuchen mit variierten Strahlungsbedingungen und verbesserten Nachweismethoden der Pigmentvorstufen erwartet.

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Physiological genetics of quantitative characters in Arabidopsis thaliana (L.) Heynh.Part 1: segregation and biosynthesis of pigments in chlorophyll-b defect mutants

Summary In biometric studies on the genetics of quantitative characters the problem of regulation of the activity of genes is rarely considered. Mutants of Arabidopsis thaliana (L.) Heynh. (Cruciferae), defective for chlorophyll b, permit a direct biochemical and physiological determination of their quantitative gene effects in the biosynthetic pathway of the plastid pigments. In agreement with other authors, the detectable genotypes are found to be based on multiple alleles (ch ⁺, ch ¹ and ch ²) at the ch locus. This evidence was obtained by test crosses of two mutants with the wild type and by a phaenotypic classification of the progeny on the basis of preparatory thin layer chromatography, paper chromatography tests of single plants, and quantitative spectrophotometry of the lines. In ch ¹/ch ¹ the synthesis of chlorophyll b is completely blocked, in ch ²/ch ² only about 10 percent of the wild-type pigment is present, and in the heterozygote ch ²/ch ¹ a complementation effect is observed (i. e. superdominance of ch ² over ch ¹) resulting in still more pigment production. In the segregating generations the variances for chlorophyll and carotinoids of all the genotypes are higher than in the parents and therefore suggest a possible polygenic background for pigment development. However, the physiological effect of the genetic background is slight as compared with the effect of the major gene ch. Preliminary experiments show that in tissue extracts of wild type leaves (ch ⁺/ch ⁺) there is an effective agent which enables irradiated chloroplasts of the defective ch ¹/ch ¹ and ch ²/ch ² genotypes to synthesize chlorophyll b in vitro. Further information on the mechanism, extent and the time of action of the active agent should be obtained by varying the conditions of irradiation and by refining the tests for pigment precursors.

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Die pigmentphysiologischen Untersuchungen wurden durch eine. Sachbeihilfe der Deutschen Forschungsgemeinschaft unterstützt.     

Abscisic acid deficiency prevents development of freezing tolerance in Arabidopsis thaliana (L.) Heynh.

Summary Abscisic acid (ABA) has been implicated as a regulatory factor in plant cold acclimation. In the present work, the cold-acclimation properties of an ABA-deficient mutant (aba) of Arabidopsis thaliana (L.) Heynh. were analyzed. The mutant had apparently lost its capability to cold acclimate: the freezing tolerance of the mutant was not increased by low temperature treatment but stayed at the level of the nonacclimated wild type. The mutational defect could be complemented by the addition of exogenous ABA to the growth medium, restoring freezing tolerance close to the wild-type level. This suggests that ABA might have a central regulatory function in the development of freezing tolerance in plants. Cold acclimation has been previously correlated to the induction of a specific set of proteins that have been suggested to have a role in freezing tolerance. However, these proteins were also induced in the aba mutant by low temperature treatment.     

Gravitropism in leaves of Arabidopsis thaliana (L.) Heynh

In higher plants, stems and roots show negative and positive gravitropism, respectively. However, current knowledge on the graviresponse of leaves is lacking. In this study, we analyzed the positioning and movement of rosette leaves of Arabidopsis thaliana under light and dark conditions. We found that the radial positioning of rosette leaves was not affected by the direction of gravity under continuous white light. In contrast, when plants were shifted to darkness, the leaves moved upwards, suggesting negative gravitropism. Analysis of the phosphoglucomutase and shoot gravitropism 2-1 mutants revealed that the sedimenting amyloplasts in the leaf petiole are important for gravity perception, as is the case in stems and roots. In addition, our detailed physiological analyses revealed a unique feature of leaf movement after the shift to darkness, i.e. movement could be divided into negative gravitropism and nastic movement. The orientation of rosette leaves is ascribed to a combination of these movements.     

Ion channels in guard cells of Arabidopsis thaliana (L.) Heynh.

Despite the availability of many mutants for signal transduction, Arabidopsis thaliana guard cells have so far not been used in electrophysiological research. Problems with the isolation of epidermal strips and the small size of A. thaliana guard cells were often prohibiting. In the present study these difficulties were overcome and guard cells were impaled with double-barreled microelectrodes. Membrane-potential recordings were often stable for over half an hour and voltage-clamp measurements could be conducted. The guard cells were found to exhibit two states. The majority of the guard cells had depolarized membrane potentials, which were largely dependent on external K⁺ concentrations. Other cells displayed spontaneous transitions to a more hyperpolarized state, at which the free-running membrane potential (E_m) was not sensitive to the external K⁺ concentration. Two outward-rectifying conductances were identified in cells in the depolarized state. A slow outward-rectifying channel (s-ORC) had properties resembling the K⁺-selective ORC of Vicia faba guard cells (Blatt, 1988, J Membr Biol 102: 235–246). The activation and inactivation times and the activation potential, all depended on the reversal potential (E_rev) of the s-ORC conductance. The s-ORC was blocked by Ba²⁺ (K_1/2 = 0.3–1.3mM) and verapamil (K_1/2 = 15–20 μM). A second rapid outward-rectifying conductance (r-ORC) activated instantaneously upon stepping the voltage to positive values and was stimulated by Ba²⁺. Inward-rectifying channels (IRC) were only observed in cells in the hyperpolarized state. The activation time and activation potential of this channel were not sensitive to the external K⁺ concentration. The slow activation of the IRC (t_1/2 ≈ 0.5 s) and its negative activation potential (V_threshold = −155 mV) resemble the values found for the KAT1 channel expressed in Saccharomyces cerevisiae (Bertl et al., 1995, Proc Natl Acad Sci USA 92: 2701–2705). The results indicate that A. thaliana guard cells provide an excellent system for the study of signal transduction processes. Received: 28 March 1996 / Accepted: 11 November 1996     

拟南芥钾离子吸收、转运及低钾胁迫的分子机理研究进展

钾(K)作为植物所需的3种大量元素之一,参与体内诸多的生理和生化过程,对于植物的生长和发育极其重要。目前,国内外学者对植物吸收、运输和利用K⁺的研究已有一定深度,尤其以模式植物拟南芥(Arabidopsis thaliana(L.) Heynh.)为研究对象。其中,与K⁺吸收、转运相关的离子通道和转运蛋白一直都是研究热点。本文综合近年来国内外相关研究进展,主要阐述K⁺通道和转运蛋白,K⁺的吸收和运输,类钙调磷酸酶(Calcineurin B-Like, CBL)-CBL相互作用蛋白激酶(CBL-Interacting protein kinase, CIPK)信号途径,参与该信号转导的一些小信号分子,对K⁺研究方面存在的问题进行了总结,并对未来的研究方向进行了展望。     

Catalase is encoded by a multigene family in Arabidopsis thaliana (L.) Heynh.

The catalase multigene family in Arabidopsis includes three genes encoding individual subunits that associate to form at least six isozymes that are readily resolved by nondenaturing gel electrophoresis. CAT1 and CAT3 map to chromosome 1, and CAT2 maps to chromosome 4. The nucleotide sequences of the three coding regions are 70 to 72% identical. The amino acid sequences of the three catalase subunits are 75 to 84% identical and 87 to 94% similar, considering conservative substitutions. Both the individual isozymes and the individual subunit mRNAs show distinct patterns of spatial (organ-specific) expression. Six isozymes are detected in flowers and leaves and two are seen in roots. Similarly, mRNA abundance of the three genes varies among organs. All three mRNAs are highly expressed in bolts, and CAT2 and CAT3 are highly expressed in leaves.     

Sodium transport by endocytic vesicles in cultured Arabidopsis thaliana (L.) Heynh. cells

In Vitro Cellular & Developmental Biology - Plant - The involvement of endocytosis in Na+ internalization by suspension-cultured Arabidopsis thaliana (L.) Heynh. cells under salt stress was...     

Hormone-response mutants of Arabidopsis thaliana (L.) Heynh. impaired in somatic embryogenesis

Plant hormones are considered to be the key factors involved in triggering in vitro induced plant morphogenesis, including somatic embryogenesis (SE). Mutants affected in SE and altered in hormonal response therefore provide valuable material for genetic research on in vitro induced plant embryogenesis. The capacity for SE was studied in 27 mutants with defects in response to different plant hormones: auxin, ABA, gibberellin and cytokinin, and evaluated in 2-week-old mutant and wild-type cultures in terms of their efficiency and productivity. SE was induced in vitro via a direct morphogenic pathway, through the culture of immature zygotic embryos on standard solid medium with 5 μM 2,4-D. The majority of the analyzed mutants displayed a significantly impaired capacity for SE; and those affected belonged to several different hormone-defective groups, including forms affected in auxin (axr4), gibberellin (ga) and ABA (abi, hyl1, cpb20, abh1) response. These mutants showed a significant decrease in embryogenic response as manifested by a low efficiency and/or productivity of SE. Additionally, SE efficiency was analyzed for axr4-1 mutant on media supplemented with different auxins while GA₃ and inhibitors of gibberellins (uniconazol P and paclobutrazol), were applied for pkl1-1-mutant. The selected mutants provide a valuable research tool for studying the molecular mechanisms determining the induction of embryogenesis in cultures of somatic tissues. Their usefulness in further studies is discussed.     

Population-level performance of Arabidopsis thaliana (L.) Heynh in dense monocultures

尽管几乎所有的农业物种在育种过程中都经历了从多物种环境向密集单种农田的过渡,但对非农业植物物种在单一栽培条件下的表现知之甚少。鉴于最近的研究成果突出杂草物种拟南芥为作物育种提供新视角的可能性,本研究的目的是了解播种密度对拟南芥在不同光周期环境下群体和个体表现的影响。我们研究了拟南芥Cvi-0生态型的表现。Cvi-0的选择基于一项初步实验,发现Cvi-0生态型植物表现出强烈的竞争性能。播种密度处理设置为每株17.6、8.8、4.4、2.2和1.1 cm²,光周期环境为日照12小时或16小时。结果表明,植物在所有密度处理下均获得了恒定的总种子产量。播种密度和日照时间处理会有一些相互作用：在高播种密度和长日照时间的处理下,种子重量更大,而日照时间短的处理下种子重量与密度呈负相关。这些结果阐明了一年生植物在面对激烈的种内竞争时能够采取不同的策略,并有助于为提高群体表现的作物育种提供新的前景。     

pho3: a phosphorus-deficient mutant of Arabidopsis thaliana (L.) Heynh

A novel P-deficient mutant of Arabidopsis thaliana, pho3, was isolated by screening for root acid phosphatase (APase) activity in plants grown under low-P conditions. pho3 had 30% less APase activity in roots than the wild type and, in contrast to wild-type plants, root APase activity did not increase in response to growth in low P. However, shoot APase activity was higher in pho3 than in the wild-type plants. In addition, the pho3 mutant had a P-deficient phenotype, even when grown in P-sufficient conditions. The total P content of 11-d-old pho3 plants, grown in agar media with a plentiful supply of P, was about 25% lower than the wild-type level in the shoot, and about 65% lower in the roots. In the rosette leaves of mature soil-grown pho3 plants the total P content was again reduced, to about 50% of wild-type levels. pho3 exhibited a number of characteristics normally associated with low-P stress, including severely reduced growth, increased anthocyanin content (at least 100-fold greater than the wild type in soil-grown plants) and starch accumulation. The results suggest that the mutant is unable to respond to low internal P levels, and may lack a transporter or a signalling component involved in regulating P nutrition. Received: 21 March 2000 / Accepted: 15 August 2000     

The use of telotrisomics for centromere mapping in Arabidopsis thaliana (L.) Heynh.

For 4 different telotrisomics of Arabidopsis thaliana segregation ratios are given for a number of genes located on the extra telocentric chromosome. These data are discussed in relation to the meiotic behaviour expected with telotrisomics. By relating the frequencies of recombinants in the progeny of heterozygous telotrisomics with the map position of the loci, the centromere position may be determined (not only in respect to order, but also in respect to distance). Chromosome arms may differ with respect to univalent loss and the amount of recombination suppression near the centromere in telotrisomics.     

Histological analysis of adventitious rooting in Arabidopsis thaliana (L.) Heynh seedlings

ABSTRACT

Adventititous rooting is essential for the post-embryonic growth of the root apparatus in various species. In Arabidopsis thaliana, adventitious rooting has been reported in some mutants, and auxin seems to be the inducer of the process. The objective of the study was to identify the tissues involved in adventitious rooting in the most commonly used ecotypes for molecular and genetic studies (i.e. Columbia, Wassilewskija and Landsberg erecta) both in the presence and absence of exogenous auxin. Seedlings of the three ecotypes were grown under various conditions. When grown under 16 hours light/day for 11 days, all seedlings showed adventitious roots, both with and without auxin, however, both adventitious and lateral rooting were enhanced by exogenous auxin (2 µM naphthaleneacetic acid). Independently of the presence of auxin and of the ecotype, the hypocotyl pericycle produced adventitious roots directly (i.e., according to the same pattern of lateral root formation by the pericycle cells in the primary root). However, in the presence of auxin, roots of indirect origin also, and mainly, formed and their formation was preceded by the exfoliation of the tissues external to the stele. Exfoliation was caused by cell hypertrophy, separation, and disintegration, which mainly involved the endodermis. At the exfoliation site, the pericycle, with a minor contribution of a few endodermal cells, produced the callus from which indirect roots arose. The finding that adventitious rooting occurs in the absence of auxin (all ecotypes) indicates that this process is part of the normal root apparatus in Arabidopsis, with the hypocotyl pericycle as the target tissue of the process. Exogenous auxin alters adventitious rhizogenesis mainly affecting the endodermis response.