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Mutations in two independent genes lead to suppression of the shoot apical meristem in maize.

The shoot apical meristem (SAM), initially formed during embryogenesis, gives rise to the aboveground portion of the maize (Zea mays) plant. The shootless phenotype (sml) described here is caused by disruption of SAM formation due to the synergistic interaction of mutations at two genetic loci. Seedlings must be homozygous for both sml (shootmeristemless), and the unlinked dgr (distorted growth) loci for a SAM-less phenotype to occur. Seedlings mutant only for sml are impaired in their morphogenesis to different extents, whereas the dgr mutation alone does not have a recognisable phenotype. Thus, dgr can be envisaged as being a dominant modifier of sml and the 12 (normal):3 (distorted growth):1 (shoot meristemless) segregation observed in the F(2) of the double heterozygote is the result of the interaction between the sml and dgr genes. Other segregation patterns were also observed in the F(2), suggesting instability of the dgr gene. Efforts to rescue mutant embryos by growth on media enriched with hormones have been unsuccessful so far. However, mutant roots grow normally on medium supplemented with kinetin at a concentration that suppresses wild-type root elongation, suggesting possible involvement of the mutant in the reception or transduction of the kinetin signal or transport of the hormone. The shootless mutant appears to be a valuable tool with which to investigate the organization of the shoot meristem in monocots as well as a means to assay the origins and relationships between organs such as the scutellum, the coleoptile, and leaves that are initiated during the embryogenic process.

Adenine↗

Localization of the auxin permease AUX1 suggests two functionally distinct hormone transport pathways operate in the Arabidopsis root apex.

Auxins represent an important class of plant hormone that regulate plant development. Plants use specialized carrier proteins to transport the auxin indole-3-acetic acid (IAA) to target tissues. To date, efflux carrier-mediated polar auxin transport has been assumed to represent the sole mode of long distance IAA movement. Localization of the auxin permease AUX1 in the Arabidopsis root apex has revealed a novel phloem-based IAA transport pathway. AUX1, asymmetrically localized to the plasma membrane of root protophloem cells, is proposed to promote the acropetal, post-phloem movement of auxin to the root apex. MS analysis shows that IAA accumulation in aux1 mutant root apices is impaired, consistent with an AUX1 phloem unloading function. AUX1 localization to columella and lateral root cap tissues of the Arabidopsis root apex reveals that the auxin permease regulates a second IAA transport pathway. Expression studies using an auxin-regulated reporter suggest that AUX1 is necessary for root gravitropism by facilitating basipetal auxin transport to distal elongation zone tissues.

Arabidopsis↗

A root-specific iron-regulated gene of tomato encodes a lysyl-tRNA-synthetase-like protein.

The tomato mutant chloronerva exhibits a defect in iron-uptake regulation. Despite high apoplastic and symplastic iron concentrations, the mutant shows characteristic symptoms of iron deficiency. Using a subtractive-hybridisation approach, we have screened for cDNA clones specific for genes with altered expression in wild-type versus mutant root tissue. Based on this clone collection, we have isolated and characterised a 2075-bp full-length cDNA encoding a lysyl-tRNA-synthetase-like protein. The corresponding gene is localised as a single copy on chromosome 10. Its expression is strongly induced by changes in the iron status of the plant. This iron-dependent regulation is superimposed upon a strict root specificity of gene expression. Possible functions of the gene product other than in protein biosynthesis will be discussed.

Amino Acid Sequence↗

Mechanisms of cytoplasmic pH regulation in hypoxic maize root tips and its role in survival under hypoxia.

We show that a transient lactic fermentation provides the signal triggering ethanol production in hypoxic maize root tips. The signal is cytoplasmic pH. This interaction between lactic and ethanolic fermentation permits tight cytoplasmic pH regulation during hypoxia--cytoplasmic pH remaining near neutrality for several hours. Mutant roots unable to synthesize ethanol can neither regulate cytoplasmic pH nor maintain ATP levels during extended periods of hypoxia and, like vertebrate tissues, are less tolerant of hypoxia than normal maize. This indicates that cytoplasmic pH regulation is an important factor in survival under hypoxia.

Cytoplasm↗

Structural studies of alfalfa roots infected with nodulation mutants of Rhizobium meliloti.

Alfalfa roots infected with four nodulation defective (Nod-) mutants of Rhizobium meliloti which were generated by transposon Tn5 mutagenesis were examined by light and electron microscopy. In one class of Nod- mutants, which we can nonreactive, the bacteria did not induce root hair curling or penetrate host cells. In a second class of Nod- mutants, which we call reactive, the bacteria induced some root hair curling and entered root epidermal cells, although no infection threads were formed. In addition, reactive Nod- mutants induced extensive root hair proliferation and hypertrophied roots. This study presents the details of the phenotype of the association between each mutant strain and alfalfa roots.

Medicago sativa↗

Plant development: pulled up by the roots.

Recent advances in the study of root development in Arabidopsis have begun to yield mechanistic insights into the processes that underpin morphogenesis, pattern formation and cell differentiation in plants. The most exciting feature of these advances is that, as a consequence of the simple and largely invariant cellular architecture of the root, the processes can be studied at a cellular level. Nevertheless, it is clear that although the cell lineages of the root are relatively invariant, we have no evidence that lineage per se is an important regulator of development. Instead, all the evidence indicates that the positional regulation of inductive cues is of primary importance. The availability of new root mutants is alerting us to the complexity of the contribution of cell size and cell expansion to plant development.

Arabidopsis↗

A pleiotropic Arabidopsis thaliana mutant with inverted root chirality.

Circumnutation is an oscillating movement of a growing plant organ that is believed to result from an endogenous rhythmic process intrinsic to growth. Circumnutating organs, as they extend, describe a helical trace. In Arabidopsis thaliana (L.) Heynh. circumnutation is particularly evident in primary roots and occurs, as in most plants, in a right-handed direction when viewed from above in the direction of the growing tips. We have discovered a pleiotropic mutant of Arabidopsis with left-handed root circumnutation. Major abnormalities of the mutant are: (i) a reduced size of all organs, mainly due to a defect in cell elongation or expansion; (ii) a zigzagging pattern of stem pith cells, reminiscent of the "erectoides" phenotype of the lk mutant of Pisum; (iii) roots of the mutant are gravitropic but as they grow, they form tight, left-handed coils. Genetically, the mutant depends on the presence of two independent monogenic recessive factors acting additively. The mutant alleles of both factors alter the growth of the aerial organs in a similar manner but differ at the root level: one mainly produces non-circumnutating roots, the other changes the direction of circumnutation from right to left hand.

Arabidopsis↗

[Identification of a gene, included in control of root system development in Arabidopsis thaliana].

Genetic and molecular genetic analysis of a lethal root mutant of Arabidopsis thaliana was carried out. The mutant was obtained from a collection created earlier by means of insertion mutagenesis. The mutation was found to be recessive. It was caused by an insertion of the T region of vector pLD3 used for transformation of germinating seeds when creating the collection of insertion mutants. A 118-bp DNA fragment flanking the left border of the insertion was isolated using the TAIL PCR technique, and its nucleotide sequence was determined. Computer analysis of this DNA region demonstrated that it was located in exon 32 of the YUP8HI2R.44 gene in chromosome 1.

Arabidopsis↗

Cell biology of plant gravity sensing.

The debate about whether gravity sensing relies upon statoliths (amyloplasts that sediment) has intensified with recent findings of gravitropism in starchless mutants and of claims of hydrostatic gravity sensing. Starch and significant plastid sedimentation are not necessary for reduced sensing in mutant roots, but plastids might function here if there were a specialized receptor for plastid mass e.g. in the ER. Alternatively, components in addition to amyloplasts might provide mass for sensing. The nucleus is dense and its position is regulated, but no direct data exist for its role in sensing. If the weight of the protoplast functioned in sensing, why would there be specific cytological specializations favoring sedimentation rather than cell mass? Gravity has multiple effects on plants in addition to gravitropism. There may be more than one mechanism of gravity sensing.

Cell Nucleus↗

The growth defect of lrt1, a maize mutant lacking lateral roots, can be complemented by symbiotic fungi or high phosphate nutrition.

The growth of three maize (Zea mays L.) mutants, each impaired in the formation of one individual element of its root system, was compared under "natural" limiting phosphate conditions (0.1 mM). Mutant plants exhibiting a reduction in root hairs (rth3-1) or a depletion of crown and brace roots (rtcs) grew as well as the corresponding wild-type plants. However, mutant plants lacking lateral roots (lrt1) showed a strong reduction in plant growth. The growth defect of lrt1 was overcome when it was grown in association with an arbuscular mycorrhizal fungus, Glomus mosseae. Establishment of symbiosis was associated with the occurrence of a new type of lateral root. These new lateral roots were stunted and highly branched, giving rise to a bush-like structure. Supply of high phosphate (1 microM) ameliorated the growth of lrt1 plants too, but less efficiently than the symbiosis did. Hence, arbuscular mycorrhizal fungi as well as phosphate functionally complemented the lrt1 mutation.

Fungi↗

Reduced gravitropism in inflorescence stems and hypocotyls, but not roots, of Arabidopsis mutants with large plastids.

The sites of gravity perception are columella cells in roots and endodermal cells in hypocotyls and inflorescence stems. Since plastids are likely to play a role in graviperception, we investigated gravitropism in plastid mutants of Arabidopsis. Previous studies have shown that the arc6 and arc12 (accumulation and replication of chloroplasts) mutants have an average of two large plastids per leaf mesophyll cell. In this study, we found that these arc mutants have altered plastid morphology throughout the entire plant body, including the cells involved in gravity perception. There were no major differences in total starch content per cell in endodermal and columella cells of the wild-type (WT) compared to arc6 and arc12 as assayed by iodine staining. Thus, the total mass of plastids per cell in arc6 and arc12 is similar to their respective WT strains. Results from time course of curvature studies demonstrated that the plastid mutation affected gravitropism only of inflorescence stems and hypocotyls, but not roots. Thus, roots appear to have different mechanisms of gravitropism compared to stems and hypocotyls. Time course of curvature studies with light-grown seedlings were performed in the presence of latrunculin B (Lat-B), an actin-depolymerizing drug. Lat-B promoted gravitropic curvature in hypocotyls of both the WT and arc6 but had little or no effect on gravitropism in roots of both strains. These results suggest that F-actin is not required for hypocotyl gravitropism.

Journal Article↗

A non-nodulating alfalfa mutant displays neither root hair curling nor early cell division in response to Rhizobium meliloti.

The early events in the alfalfa-Rhizobium meliloti symbiosis include deformation of epidermal root hairs and the approximately concurrent stimulation of cell dedifferentiation and cell division in the root inner cortex. These early steps have been studied previously by analysis of R. meliloti mutants. Bacterial strains mutated in nodABC, for example, fail to stimulate either root hair curling or cell division events in the plant host, whereas exopolysaccharide (exo) mutants of R. meliloti stimulate host cell division but the resulting nodules are uninfected. As a further approach to understanding early symbiotic interactions, we have investigated the phenotype of a non-nodulating alfalfa mutant, MnNC-1008 (NN) (referred to as MN-1008). Nodulating and non-nodulating plants were inoculated with wild-type R. meliloti and scored for root hair curling and cell divisions. MN-1008 was found to be defective in both responses. Mutant plants inoculated with Exo- bacteria also showed no cell division response. Therefore, the genetic function mutated in MN-1008 is required for both root hair curling and cell division, as is true for the R. meliloti nodABC genes. These observations support the model that the distinct cellular processes of root hair curling and cell division are triggered by related mechanisms or components, or are causally linked.

Cell Division↗

The central-peripheral transition zone of cervical spinal nerve roots in Jimpy mutant and normal mice. Light- and electron-microscopic study.

Comparative morphological and ultrastructural investigations on the cervical dorsal and ventral central-peripheral transition zones (CPTZs) of Jimpys and control mice have been performed at early and advanced myelination stages. After postnatal development a characteristic cone-shaped glial outgrowth extends into the proximal part of the dorsal roots, while the ventral roots exhibit short Schwann cell and peripheral nervous tissue invaginations into the spinal cord at the ventral root-spinal cord junction in both animal groups. In Jimpys, although there is marked central myelin deficiency and absence of oligodendroglial development on the CNS side, the normal general aspect of the CPTZs is maintained. Previously postulated astrocytic and neuroaxonal abnormalities in the mutants do not alter the central-peripheral borderline, and Schwann cell migration from the spinal nerve roots into the cord does not occur.

Animals↗

Effects of Rhizobium meliloti nif and fix mutants on alfalfa root nodule development.

Ineffective alfalfa nodules were examined at the light and electron microscope level after inoculation with Rhizobium meliloti strains with mutations in nif and fix genes. All the mutant strains induced nodules that contained elongated bacteroids within the host cells, but the bacteroids quickly senesced. The nodules were small and numerous, and the host cells also exhibited symptoms of an ineffective symbiosis. nifB, fixA, and fixB bacteroids appeared to be completely differentiated (by ultrastructural criteria), i.e., as bacteroids developed, they increased in diameter and length and their cytoplasm underwent a change from homogeneous and electron dense to heterogeneous and electron transparent after enlargement. In contrast, nifA bacteroids rarely matured to this state. The bacteroids degenerated at an earlier stage of development and did not become electron transparent.

Medicago sativa↗

Reduced gravitropic sensitivity in roots of a starch-deficient mutant of Nicotiana sylvestris.

Gravitropism was studied in seedlings of Nicotiana sylvestris Speg. et Comes wild-type (WT) and mutant NS 458 which has a defective plastid phosphoglucomutase (EC 2.7.5.1.). Starch was greatly reduced in NS 458 compared to the WT, but small amounts of starch were detected in rootcap columella cells in NS 458 by light and electron microscopy. The roots of WT are more sensitive to gravity than mutant NS 458 roots since: (1) in mutant roots, curvature was reduced and delayed in the time course of curvature; (2) curvature of mutant roots was 24-56% that of WT roots over the range of induction periods tested; (3) in intermittent-stimulation experiments, curvature of mutant roots was 37% or less than that of WT roots in all treatments tested. The perception time, determined by intermittent-stimulation experiments, was < or = 5 s for WT roots and 30-60 s for mutant roots. The growth rates for WT and NS 458 roots were essentially equal. These results and our previous results with WT and starchless mutant Arabidopsis roots (Kiss et al. 1989, Planta 177, 198-206) support the conclusions that a full complement of starch is necessary for full gravitropic sensitivity and that amyloplasts function in gravity perception. Since a presumed relatively small increase in plastid buoyant mass (N. sylvestris mutant versus Arabidopsis mutant) significantly improves the orientation of the N. sylvestris mutant roots, we suggest that plastids are the likeliest candidates to be triggering gravity perception in roots of both mutants.

Gravitropism↗

The effects of asymmetric calcium application on primary root curvature of ageotropum pea mutant.

Recent studies indicate that roots of ageotropum seedlings can be used to study the hydrotropic response of roots independent of physiological events related to the gravity response of roots. There is evidence that Ca2+ ions are important in both the gravitropic and hydrotropic response of roots. In this study, we have compared three fully graviresponsive pea cultivars and the ageotropum mutant with regard to: 1) general root anatomy, 2) the effects of unilateral Ca application to both the root cap and DEZ region on root curvature, and 4) effects of unilateral application of EGTA to the DEZ region.

Calcium↗

Ultrastructure and movements of cell organelles in the root cap of agravitropic mutants and normal seedlings of Arabidopsis thaliana.

The root anatomy and ultrastructure of the agravitropic Arobidopsis thaliana L. mutants Dwf and aux-1 were compared with the gravitropic mutant aux-2 and the wild type (WT) in an attempt to find an explanation for the lack of response to gravity. No differences were found in the organization of the root cap. The central part of the cap (columella) contains 5 storeys of developing, functioning and degenerating statocytes. Their ultrastructure is very similar in all four types of plant. Particular attention was paid to the distribution of rough endoplasmic reticulum (ER). Both in the WT and the mutants the ER is concentrated in the distal part at the "floor" of the cell. Light micrographs were used to compare the sedimentation rates of movable cell structures in normal and agravitropic root statocytes. A longitudinal movement of amyloplasts and nuclei was observed when the roots were inverted. In WT and aux-2 the rates were on average 6.3 micrometers h-1 (amyloplasts) and 2.1 micrometers h-1 (nucleus). In aux-1 the sedimentation rates were significantly lower: 2.4 and 0.6 micrometers h-1, respectively. Based on magnified electron micrographs of normal and inverted statocytes a morphometrical analysis of the distribution and redistribution of amyloplasts, nuclei, mitochondria, vacuoles and ER was made. The only significant difference was found in the redistribution of amyloplasts between aux-1 and the gravitropical normal types.

Arabidopsis↗