Effect of gravity on apical dominance in Pharbitis nil.

When the upper part of main shoot of morning glory (Pharbitis nil) is gently bent down, lateral bud on the bending region is released from apical dominance and starts to elongate. But, clinorotating the bending shoots prevents the release of the lateral bud from apical dominance. These results suggest that gravity affects apical dominance in morning glory. Here we verified the gravity-regulated apical dominance by using a weeping morning glory defective in gravitropic response due to abnormal differentiation of endodermis. That is, bending main shoot of the weeping morning glory hardly caused the lateral bud to elongate. In addition, decapitation of apical bud released the lateral bud from apical dominance, and exogenous auxin applied to the cut surface of the decapitated stem was inhibitory to the outgrowth of the lateral bud in the wild type. However, the effect of auxin was much less in the weeping morning glory. Thus, apical dominance of the weeping morning glory was weaker and less influenced by gravity than that of the wild type, which could occur due to abnormal differentiation of endodermis required for graviperception.     

Interaction of light and gravitropism with nutation of hypocotyls of Arabidopsis thaliana seedlings

Etiolated seedlings of Arabidopsis thaliana nutated under conditions of physiological darkness while about ten percent of monitored individuals exhibited regular elliptical nutation, circumnutation. Pre-irradiation with red light prevented occurrence of circumnutation without having an effect on the average rate of the nutational movement. Phototropic response of seedlings to unilateral blue light appeared to be superimposed over nutation. Throughout gravitropism, some seedlings continued to exhibit nutation suggesting that these two processes are independently controlled. Based on these results, we suggest that nutation in Arabidopsis probably is not controlled by the mechanism predicted by the theory of gravitropic overshoots.     

SGR1, SGR2, SGR3: novel genetic loci involved in shoot gravitropism in Arabidopsis thaliana.

In higher plants shoots show a negative gravitropic response but little is known about its mechanism. To elucidate this phenomenon, we have isolated a number of mutants with abnormal shoot gravitropic responses in Arabidopsis thaliana. Here we describe mainly three mutants: sgr1-1, sgr2-1, and sgr3-1 (shoot gravitropism). Genetic analysis confirmed that these mutations were recessive and occurred at three independent loci, named SGR1, SGR2, and SGR3, respectively. In wild type, both inflorescence stems and hypocotyls show negative gravitropic responses. The sgr1-1 mutants showed no response to gravity either by inflorescence stems or by hypocotyls. The sgr2-1 mutants also showed no gravitropic response in inflorescence stems but showed a reduced gravitropic response in hypocotyls. In contrast, the sgr3-1 mutant was found to have reduced gravitropic responses in inflorescence stems but normal gravitropic responses in hypocotyls. These results suggest that some genetic components of the regulatory mechanisms for gravitropic responses are common between inflorescence stems and hypocotyls, but others are not. In addition, these sgr mutants were normal with respect to root gravitropism, and their inflorescence stems and hypocotyls could carry out phototropism. We conclude that SGR1, SGR2, and SGR3 are novel genetic loci specifically involved in the regulatory mechanisms of shoot gravitropism in A. thaliana.     

Amyloplast displacement is necessary for gravisensing in Arabidopsis shoots as revealed by a centrifuge microscope

The starch‐statolith hypothesis proposes that starch‐filled amyloplasts act as statoliths in plant gravisensing, moving in response to the gravity vector and signaling its direction. However, recent studies suggest that amyloplasts show continuous, complex movements in Arabidopsis shoots, contradicting the idea of a so‐called ‘static’ or ‘settled’ statolith. Here, we show that amyloplast movement underlies shoot gravisensing by using a custom‐designed centrifuge microscope in combination with analysis of gravitropic mutants. The centrifuge microscope revealed that sedimentary movements of amyloplasts under hypergravity conditions are linearly correlated with gravitropic curvature in wild‐type stems. We next analyzed the hypergravity response in the shoot gravitropism 2 (sgr2) mutant, which exhibits neither a shoot gravitropic response nor amyloplast sedimentation at 1  g . sgr2 mutants were able to sense and respond to gravity under 30  g conditions, during which the amyloplasts sedimented. These findings are consistent with amyloplast redistribution resulting from gravity‐driven movements triggering shoot gravisensing. To further support this idea, we examined two additional gravitropic mutants, phosphoglucomutase (pgm) and sgr9, which show abnormal amyloplast distribution and reduced gravitropism at 1  g . We found that the correlation between hypergravity‐induced amyloplast sedimentation and gravitropic curvature of these mutants was identical to that of wild‐type plants. These observations suggest that Arabidopsis shoots have a gravisensing mechanism that linearly converts the number of amyloplasts that settle to the ‘bottom’ of the cell into gravitropic signals. Further, the restoration of the gravitropic response by hypergravity in the gravitropic mutants that we tested indicates that these lines probably have a functional gravisensing mechanism that is not triggered at 1  g .     

How do plant shoots bend up? The initial step to elucidate the molecular mechanisms of shoot gravitropism using Arabidopsis thaliana

In higher plants, shoots show a negative gravitropic response. To elucidate the molecular mechanisms of this phenomenon, mutational analyses using Arabidopsis thaliana are in progress. This minireview aims to present recent developments in the genetic analysis of shoot gravitropism in this organism. We focus mainly on our studies on the novel shoot gravitropic (sgr) mutants in Arabidopsis thaliana that have dramatic defects in shoot gravitropism.     

The gravity-regulated growth of axillary buds is mediated by a mechanism different from decapitation-induced release

When the upper part of the main shoot of the Japanese morning glory (Pharbitis nil or Ipomoea nil) is bent down, the axillary bud situated on the uppermost node of the bending region is released from apical dominance and elongates. Here, we demonstrate that this release of axillary buds from apical dominance is gravity regulated. We utilized two agravitropic mutants of morning glory defective in gravisensing cell differentiation, weeping (we) and weeping2 (we2). Bending the main shoots of either we or we2 plants resulted in minimal elongation of their axillary buds. This aberration was genetically linked to the agravitropism phenotype of the mutants, which implied that shoot bending-induced release from apical dominance required gravisensing cells. Previous studies have shown that basipetal translocation of auxin from the apical bud inhibits axillary bud growth, whereas cytokinin promotes axillary bud outgrowth. We therefore compared the roles of auxin and cytokinin in bending- or decapitation-induced axillary bud growth. In the wild-type and we plants, decapitation increased cytokinin levels and reduced auxin response. In contrast, shoot bending did not cause significant changes in either cytokinin level or auxin response, suggesting that the mechanisms underlying gravity- and decapitation-regulated release from apical dominance are distinct and unique.     

The endodermis and shoot gravitropism

Shoots and roots of higher plants exhibit negative and positive gravitropism, respectively. A variety of gravitropic mutants have recently been isolated from Arabidopsis, the characterization of which demonstrates that the molecular mechanisms of the gravitropic responses in roots, hypocotyls and inflorescence stems are different. The cytological and molecular analysis of two mutants, shoot gravitropism 1 (sgrl), which is allelic to scarecrow (scr), and sgr7, which is allelic to short-root(shr), indicate that the endodermis is the site of gravity perception in shoots. These data suggest a new model for shoot gravitropism.     

Root movements: Towards an understanding through attempts to model the processes involved

Roots have the ability to change the direction of their forward growth. Sometimes these directional changes are rapid, as in mutations, or they are slower, as in tropisms. The gravitational force is always present and roots have an efficient graviperception mechanism which enables them to initiate gravitropic movements. In trying to model and simulate the course of gravitropic root movements with a view to analyse the component processes, the following aspects of the plant's interaction with gravity have been considered: (1) The level of organization (organism, organ, cell) at which the movement process is expressed; (2) whether the gravity stimulation event is dynamic or static (i.e. whether or not physiologically significant displacements take place with respect to the gravity vector); (3) the sub-systems involved in movement and the processes which they regulate; (4) the mathematical characterization of the relevant sub-systems. A further allied topic is the nature of nutational movements and whether they are linked with gravitropic movements in some way. In considering how they can best be modelled, two types of nutational movements are proponed: stochastic nutation and circumnutation. Most, if not all, natural movements developed in response to static gravistimulation can be viewed as gravimorphisms. This applies at the levels of cell, organ and organism. However, when a system at any one of these levels experiences dynamic gravistimulation, because of its inherent homeostatic properties, it is induced to regenerate a state similar to that previously held. Thus, gravitropism is a regenerative gravimorphic process at the level of the organ.     

Circumnutational movement in rice coleoptiles involves the gravitropic response: analysis of an agravitropic mutant and space‐grown seedlings

Plants exhibit helical growth movements known as circumnutation in growing organs. Some studies indicate that circumnutation involves the gravitropic response, but this notion is a matter of debate. Here, using the agravitropic rice mutant lazy1 and space‐grown rice seedlings, we found that circumnutation was reduced or lost during agravitropic growth in coleoptiles. Coleoptiles of wild‐type rice exhibited circumnutation in the dark, with vigorous oscillatory movements during their growth. The gravitropic responses in lazy1 coleoptiles differed depending on the growth stage, with gravitropic responses detected during early growth and agravitropism during later growth. The nutation‐like movements observed in lazy1 coleoptiles at the early stage of growth were no longer detected with the disappearance of the gravitropic response. To verify the relationship between circumnutation and gravitropic responses in rice coleoptiles, we conducted spaceflight experiments in plants under microgravity conditions on the International Space Station. Wild‐type rice seeds were germinated, and the resulting seedlings were grown under microgravity or a centrifuge‐generated 1 g environment in space. We began filming the seedlings 2 days after seed imbibition and obtained images of seedling growth every 15 min. The seed germination rate in space was 92–100% under both microgravity and 1 g conditions. LED‐synchronized flashlight photography induced an attenuation of coleoptile growth and circumnutational movement due to cumulative light exposure. Nevertheless, wild‐type rice coleoptiles still showed circumnutational oscillations under 1 g but not microgravity conditions. These results support the idea that the gravitropic response is involved in plant circumnutation.     

Role of endodermal cell vacuoles in shoot gravitropism

In higher plants, shoots and roots show negative and positive gravitropism, respectively. Data from surgical ablation experiments and analysis of starch deficient mutants have led to the suggestion that columella cells in the root cap function as gravity perception cells. On the other hand, endodermal cells are believed to be the statocytes (that is, gravity perceiving cells) of shoots. Statocytes in shoots and roots commonly contain amyloplasts which sediment under gravity. Through genetic research with Arabidopsis shoot gravitropism mutants, sgr1/scr and sgr7/shr, it was determined that endodermal cells are essential for shoot gravitropism. Moreover, some starch biosynthesis genes and EAL1 are important for the formation and maturation of amyloplasts in shoot endodermis. Thus, amyloplasts in the shoot endodermis would function as statoliths, just as in roots. The study of the sgr2 and zig/sgr4 mutants provides new insights into the early steps of shoot gravitropism, which still remains unclear. SGR2 and ZIG/SGR4 genes encode a phospholipase-like and a v-SNARE protein, respectively. Moreover, these genes are involved in vacuolar formation or function. Thus, the vacuole must play an important role in amyloplast sedimentation because the sgr2 and zig/sgr4 mutants display abnormal amyloplast sedimentation.     

Characterization and genetic analysis of a lettuce (Lactuca sativa L.) mutant,weary, that exhibits reduced gravitropic response in hypocotyls and inflorescence stems

A lettuce (Lactuca sativa L.) mutant that exhibits a procumbent growth habit was identified and characterized. In two wild type (WT) genetic backgrounds, segregation patterns revealed that the mutant phenotype was controlled by a recessive allele at a single locus, which was designated weary. Hypocotyls and inflorescence stems of plants homozygous for the weary allele exhibited reduced gravitropic responses compared with WT plants, but roots exhibited normal gravitropism. Microscopic analysis revealed differences in the radial distribution of amyloplasts in hypocotyl and inflorescence stem cells of weary and WT plants. Amyloplasts occurred in a single layer of endodermal cells in WT hypocotyls and inflorescence stems. By contrast, amyloplasts were observed in several layers of cortical cells in weary hypocotyls, and weary inflorescence stem cells lacked amyloplasts entirely. These results are consistent with the proposed role of sedimenting amyloplasts in shoot gravitropism of higher plants. The phenotype associated with the weary mutant is similar to that described for the Arabidopsis mutant sgr1/scr, which is defective in radial patterning and gravitropism.     

Mutations in the SGR4, SGR5 and SGR6 Loci of Arabidopsis thaliana Alter the Shoot Gravitropism

Shoots of higher plants grow upward in response to gravity.To elucidate the molecular mechanism of this response, we haveisolated shoot gravitropism (sgr) mutants in Arabidopsis thaliana.In this report, we describe three novel mutants, sgr4-1, sgr5-1and sgr6-1 whose inflorescence stems showed abnormal gravitropicresponses as previously reported for sgr1, sgr2 and sgr3. Thesenew sgr mutations were recessive and occurred at three independentgenetic loci. The sgr4-1 mutant showed severe defect in gravitropismof both inflorescence stem and hypocotyl but were normal inroot gravitropism as were sgr1 and sgr2. The sgr5-1 and sgr6-1mutants showed reduced gravitropism only in inflorescence stemsbut normal in both hypocotyls and roots as sgr3. These resultssupport the hypothesis that some mechanisms of gravitropismare genetically different in these three organs in A. thaliana.In addition, these mutants showed normal phototropic responses,suggesting that SGR4, SGR5 and SGR6 genes are specifically involvedin gravity perception and/or gravity signal transduction forthe shoot gravitropic response. (Received November 21, 1996; Accepted February 17, 1997)     

Circumnutation of rice coleoptiles: its occurrence, regulation by phytochrome, and relationship with gravitropism

It has been found that coleoptiles of dark-grown rice (Oryza sativa L.) seedlings undergo regular circumnutation in circular orbits with periods of about 180 min. Both clockwise and counter-clockwise movements were observed, but individual coleoptiles continued to rotate only in one direction. Light-grown seedlings did not show circumnutation. In fact, dark-grown seedlings were found to cease circumnutating in response to a pulse of red light (R). This light-induced inhibition of circumnutation was demonstrated to involve both a FR-inducible very-low-fluence response, solely mediated by phytochrome A, and a FR-reversible low-fluence response, mediated by phytochrome B and/or C. The R-induced inhibition of circumnutation showed temporal agreement with the R-induced inhibition of coleoptile growth, suggesting that the former results from the latter. However, about 25% of growth activity remained after R treatment, indicating that circumnutation is more specifically regulated by phytochrome. The R-treated coleoptile showed gravitropism. Investigation of the growth differential for gravitropic curvature revealed that gravitropic responsiveness was rather enhanced by R. The results suggested that gravitropism is not a cause of circumnutation. It remained probable, however, that gravity perception is a part of the mechanism of circumnutation. It is speculated that the circumnutation investigated aids the seedling shoot in growing through the soil.     

SGR2, a phospholipase-like protein, and ZIG/SGR4, a SNARE, are involved in the shoot gravitropism of Arabidopsis

In higher plants, the shoot and the root generally show negative and positive gravitropism, respectively. To elucidate the molecular mechanisms involved in gravitropism, we have isolated many shoot gravitropism mutants in Arabidopsis. The sgr2 and zig/sgr4 mutants exhibited abnormal gravitropism in both inflorescence stems and hypocotyls. These genes probably are involved in the early step(s) of the gravitropic response. The sgr2 mutants also had misshapen seed and seedlings, whereas the stem of the zig/sgr4 mutants elongated in a zigzag fashion. The SGR2 gene encodes a novel protein that may be part of a gene family represented by bovine phosphatidic acid-preferring phospholipase A1 containing a putative transmembrane domain. This gene family has been reported only in eukaryotes. The ZIG gene was found to encode AtVTI11, a protein that is homologous with yeast VTI1 and is involved in vesicle transport. Our observations suggest that the two genes may be involved in a vacuolar membrane system that affects shoot gravitropism.     

Chiral and non-chiral nutations in Arabidopsis roots grown on the random positioning machine

Arabidopsis thaliana roots grown on a vertically set plate do not elongate straight down the gravitational vector, but by making waves and coils, and by conspicuously slanting towards the right-hand. This behaviour, in a previous paper, was ascribed to the simultaneous effect of three processes: circumnutation, positive gravitropism and negative thigmotropism. However, when the plants are grown on the Random Positioning Machine (RPM), in conditions that are believed to simulate space microgravitational conditions closely, the roots do not show the usual pattern. In the wild type, the roots make large loops to the right-hand side, whereas in the gravitropic and auxinic mutants aux1, eir1, rha1, they just move randomly around the initial direction. Therefore, if the movements made on the RPM are those produced by the exclusion of gravitropism and negative thigmotropism, as is apparent, the conclusion is that Arabidopsis roots are animated by a form of chiral circumnutation, that is lacking in the auxinic and gravitropic mutants aux1, eir1 and rha1. In addition, the 1 g condition appears to reduce the scatter among the circumnutating tracks produced by the roots of the wild types, but not among those of the mutants. Because there is a scarcity of literature regarding circumnutation in roots, it is not known how widely root chiral circumnutation is spread, but it is known that, in some previously studied species, just random nutations are observed. Two kinds of nutating movements seem to exist in plant roots and, whereas the random process does not seem to be connected with auxin physiology and transport, the chiral process appears to be connected in the same way as gravitropism is.     

Involvement of the vacuoles of the endodermis in the early process of shoot gravitropism in Arabidopsis

The endodermal cells of the shoot are thought to be the gravity-sensing cells in Arabidopsis. The amyloplasts in the endodermis that sediment in the direction of gravity may act as statoliths. Endodermis-specific expression of SGR2 and ZIG using the SCR promoter could complement the abnormal shoot gravitropism of the sgr2 and zig mutants, respectively. The abnormalities in amyloplast sedimentation observed in both mutants recovered simultaneously. These results indicate that both genes in the endodermal cell layer are crucial for shoot gravitropism. ZIG encodes AtVTI11, which is a SNARE involved in vesicle transport to the vacuole. The fusion protein of SGR2 and green fluorescent protein localized to the vacuole and small organelles. These observations indicate that ZIG and SGR2 are involved in the formation and function of the vacuole, a notion supported by the results of subcellular analysis of the sgr2 and zig mutants with electron microscopy. These results strongly suggest that the vacuole participates in the early events of gravitropism and that SGR2 and ZIG functions are involved.     

Genetic evidence that the endodermis is essential for shoot gravitropism in Arabidopsis thaliana

Shoots of higher plants exhibit negative gravitropism. However, little is known about the mechanism or site of gravity perception in shoots. We have identified two loci that are essential for normal shoot gravitropism in Arabidopsis thaliana . Genetic analysis demonstrated that the shoot gravitropism mutants sgr1 and sgr7 are allelic to the radial pattern mutants, scr and shr , respectively. Characterization of the aerial phenotype of these mutants revealed that the primary defect is the absence of a normal endodermis in hypocotyls and inflorescence stems. This indicates that the endodermis is essential for shoot gravitropism and strongly suggests that this cell layer functions as the gravity-sensing cell layer in dicotyledonous plant shoots. These results also demonstrate that, in addition to their previously characterized role in root radial patterning, SCR and SHR regulate the radial organization of the shoot axial organs in Arabidopsis .      

Physiology of Movements in Stems of Seedling Pisum sativum L. cv. Alaska : I. Experimental Separation of Nutation from Gravitropism

Gravitropism and nutation in the stems of dark-grown, seedling peas (Pisum sativum L. cv. Alaska) were recorded on time-lapse photographs made with photomorphogenetically inactive light. Although gravitropism and nutation have been connected by several different theories in the past, our experiments indicate that the two processes are in fact dissociable. The evidence is as follows: (a) Nutational patterns are asymmetric. There is much greater amplitude of oscillation in the plane parallel () to the plane of the apical hook than in the plane perpendicular (), yet the average gravitropic response is equal in these two planes. (b) Brief red light irradiation given 16 to 24 hours before observation greatly increases the amplitude of nutation in the -plane, but has no influence on the kinetics of gravitropic response. (c) An inhibitor of auxin transport, α-naphthylphthalamic acid, strongly inhibits nutation at 5 micromolar but affects gravitropism only at higher concentrations. (d) Nutation is also strongly inhibited by removal of the apical bud, but gravitropism is unaffected. (e) The period of nutation does not exhibit a constant relationship to the response time of gravitropism. The above evidence is inconsistent with theories that gravitropism is an asymmetrically modified nutation or, alternatively, that nutational oscillations result in a simple fashion from gravitropic overshoots. The evidence is consistent with, although not proof of, autonomous factors such as an endogenous rhythm of growth as the cause of nutation in pea stems. However, gravity and nutation do interact. Nutation in a population of seedlings can be synchronized and brought into phase by a single gravitropic induction. Furthermore, the response time and initial rate of gravitropic curvature depend to some extent on the phase of nutational curvature at which gravitropic induction is begun.