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1.
Loss-of-function mutations of ROOT HAIR DEFECTIVE3 suppress root waving, skewing, and epidermal cell file rotation in Arabidopsis
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Wild-type Arabidopsis (Arabidopsis thaliana L. Heynh.) roots growing on a tilted surface of impenetrable hard-agar media adopt a wave-like pattern and tend to skew to the right of the gravity vector (when viewed from the back of the plate through the medium). Reversible root-tip rotation often accompanies the clockwise and counterclockwise curves that form each wave. These rotations are manifested by epidermal cell file rotation (CFR) along the root. Loss-of-function alleles of ROOT HAIR DEFECTIVE3 (RHD3), a gene previously implicated in the control of vesicle trafficking between the endoplasmic reticulum and the Golgi compartments, resulted in an almost complete suppression of epidermal CFR, root skewing, and waving on hard-agar surfaces. Several other root hair defective mutants (rhd2-1, rhd4-1, and rhd6-1) did not exhibit dramatic alterations in these root growth behaviors, suggesting that a generalized defect in root hair formation is not responsible for the surface-dependent phenotypes of rhd3. However, similar alterations in root growth behavior were observed in a variety of mutants characterized by defects in cell expansion (cob-1, cob-2, eto1-1, eto2-1, erh2-1, and erh3-1). The erh2-1 and rhd3-1 mutants differed from other anisotropic cell expansion mutants, though, by an inability to respond to low doses of the microtubule-binding drug propyzamide, which normally causes enhanced left-handed CFR and right skewing. We hypothesize that RHD3 may control epidermal CFR, root skewing, and waving on hard-agar surfaces by regulating the traffic of wall- or plasma membrane-associated determinants of anisotropic cell expansion. 相似文献
2.
When stimulated to bend downward by being held at 45 degrees off vertical but unable to penetrate into agar-based media, Arabidopsis roots develop waving and looping growth patterns. Here, we demonstrate that ethylene modulates these responses. We determined that agar-containing plates sealed with low-porosity film generate abiotic ethylene concentrations of 0.1 to 0.3 microL L(-1), whereas in plates wrapped with porous tape, ethylene remains at trace levels. We demonstrate that exogenous ethylene at concentrations as low as a few nanoliters per liter modulates root waving, root growth direction, and looping but through partly different mechanisms. Nutrients and Suc modify the effects of ethylene on root waving. Thus, ethylene had little effect on temporal wave frequency when nutrients were omitted but reduced it significantly on nutrient-supplemented agar. Suc masked the ethylene response. Ethylene consistently suppressed the normal tendency for roots of Landsberg erecta to skew to the right as they grow against hard-agar surfaces and also generated righthanded petiole twisting. Furthermore, ethylene suppressed root looping, a gravity-dependent growth response that was enhanced by high nutrient and Suc availability. Our work demonstrates that cell file twisting is not essential for root waving or skewing to occur. Differential flank growth accounted for both the extreme root waving on zero-nutrient plates and for root skewing. Root twisting was nutrient-dependent and was thus strongly associated with the looping response. The possible role of auxin transport in these responses and the involvement of circadian rhythms are discussed. 相似文献
3.
Arabidopsis thaliana sku mutant seedlings show exaggerated surface-dependent alteration in root growth vector. 总被引:5,自引:0,他引:5
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Roots of wild-type Arabidopsis thaliana seedlings in the Wassilewskija (WS) and Landsberg erecta (Ler) ecotypes often grow aslant on vertical agar surfaces. Slanted root growth always occurs to the right of the gravity vector when the root is viewed through the agar surface, and is not observed in the Columbia ecotype. Right-slanted root growth is surface-dependent and does not result directly from directional environmental stimuli or gradients in the plane of skewing. We have isolated two partially dominant mutations in WS (sku1 and sku2) that show an exaggerated right-slanting root-growth phenotype on agar surfaces. The right-slanting root-growth phenotype of wild-type and mutant roots is not the result of diagravitropism or of an alteration in root gravitropism. It is accompanied by a left-handed rotation of the root about its axis within the elongation zone, the rate of which positively correlates with the degree of right-slanted curvature. Our data suggest that the right-slanting root growth phenotype results from an endogenous structural asymmetry that expresses itself by a directional root-tip rotation. 相似文献
4.
It is widely accepted that the Arabidopsis Adh (alcohol dehydrogenase) gene is constitutively expressed at low levels in the roots of young plants grown on agar media, and that the expression level is greatly induced by anoxic or hypoxic stresses. We questioned whether the agar medium itself created an anaerobic environment for the roots upon their growing into the gel. beta-Glucuronidase (GUS) expression driven by the Adh promoter was examined by growing transgenic Arabidopsis plants in different growing systems. Whereas roots grown on horizontal-positioned plates showed high Adh/GUS expression levels, roots from vertical-positioned plates had no Adh/GUS expression. Additional results indicate that growth on vertical plates closely mimics the Adh/GUS expression observed for soil-grown seedlings, and that growth on horizontal plates results in induction of high Adh/GUS expression that is consistent with hypoxic or anoxic conditions within the agar of the root zone. Adh/GUS expression in the shoot apex is also highly induced by root penetration of the agar medium. This induction of Adh/GUS in shoot apex and roots is due, at least in part, to mechanisms involving Ca2+ signal transduction. 相似文献
5.
Pandey S Monshausen GB Ding L Assmann SM 《The Plant journal : for cell and molecular biology》2008,55(2):311-322
Heterotrimeric G proteins composed of α, β and γ subunits regulate a number of fundamental processes concerned with growth and development in plants. In addition to the canonical heterotrimeric G proteins, plants also contain a small family of extra large G proteins (XLGs) that show significant similarity to the G-protein α subunit in their C-terminal regions. In this paper we show that one of the three XLG genes, XLG3 , and the Gβ subunit (AGB1) of the Arabidopsis G-protein heterotrimer are specifically involved in the regulation of a subset of root morphological and growth responses. Based on analysis of T-DNA insertional mutant phenotypes, XLG3 and AGB1 each positively regulate root waving and root skewing. Since these responses are regulated by physical as well as physiological cues, we assessed the roles of AGB1 and XLG3 in gravitropism, thigmotropism and hormonal responses. Our data show that mutants lacking either XLG3 or AGB1 genes are hypersensitive to ethylene and show growth responses consistent with alterations in auxin transport, while maintaining an essentially wild-type response to the physical cues of gravity and touch. These results suggest that XLG3 and AGB1 proteins regulate the hormonal determinants of root-waving and root-skewing responses in plants and possibly interact in a tissue-specific or signal-specific manner. Because plants harboring knockout mutations in the Gα subunit gene, GPA1 , exhibit wild-type root waving and skewing, our results may indicate that the AGB1 subunit functions in these processes without formation of a classic Gαβγ heterotrimer. 相似文献
6.
Sugar-induced adventitious roots in<Emphasis Type="Italic"> Arabidopsis</Emphasis> seedlings 总被引:3,自引:0,他引:3
The effects of sugars on root growth and on development of adventitious roots were analyzed in Arabidopsis thaliana. Seeds were sown on agar plates containing 0.0–5.0% sugars and placed vertically in darkness (DD) or under long day (LD,
16 h:8 h) conditions, so that the seedlings were constantly attached to the agar medium. In the sucrose-supplemented medium,
seedlings showed sustained growth in both DD and LD. However, only dark-grown seedlings developed adventitious roots from
the elongated hypocotyl. The adventitious roots began to develop 5 days after imbibition and increased in number until day 11.
They could, however, be initiated at any position along the hypocotyl, near the cotyledon or the primary root. They were initiated
in the pericycle in the same manner as ordinary lateral roots. Sucrose, glucose and fructose greatly stimulated the induction
of adventitious roots, but mannose or sorbitol did not. Sucrose at concentrations of 0.5–2.0% was most effective in inducing
adventitious roots, although 5.0% sucrose suppressed induction. Direct contact of the hypocotyl with the sugar-supplemented
agar medium was indispensable for the induction of adventitious roots.
Electronic Publication 相似文献
7.
8.
Zea mays L. seedlings, grown on agar plates at 26°C, reoriented the original vertical direction of their primary root when exposed to a thermal gradient applied perpendicular to the gravity vector. The magnitude and direction of curvature can not be explained simply by either a temperature or a humidity effect on root elongation. It is concluded that primary roots of maize sense temperature gradients in addition to sensing the gravitational force. 相似文献
9.
一种筛选拟南芥突变体的有效方法 总被引:15,自引:0,他引:15
经甲基磺酸乙酯(EMS)诱变处理的拟南芥种子,接种于MS培养基上,垂直放置培养4天后,将幼苗转移至胁迫培养基中,以倒置幼苗180°所形成的弯曲生长根作为指标筛选拟南芥耐营养胁迫突变体。利用这种方法,成功地筛选到一个耐低钾的隐性单基因拟南芥突变体。本方法同样适用于其他类型突变体的筛选。
Abstract:his paper introduces a root-bending assay for isol ation of Arabidopsis mutants tolerant to nutrition stress. Seeds of wild-ty pe Arabidopsis thaliana (ecotype Landersberg erecta) were mutagenized wi th ethyl methyl sulfide (EMS),and M2 populations were screened for mutants. Fo ur-day-old seedlings with 1-to 1.5-cm-long roots were transferred from the vertical agar plates onto to a second agar medium that was supplemented with det erminate stress. The seedlings were arranged in rows, and the plates were orient ed vertically with the roots pointing upward. After another 4 days, the root be nding seedlings were selected for putative mutants and transferred to soil to gr ow to maturity.Seeds from the putative mutants were screened again to determine the true mutants.By using this root-bending assay we have isolated a low-K+-tolerant (lkt1) mutant which is caused by single recessive nuclear mutation. F or lkt1 mutant screening,K+concentration of the medium was 100μmol/L because root growth of wild type seedlings was completely inhibited at or below this con centration.This root-bending assay is also applicable to other type of Arabid opsis mutant isolation. 相似文献
10.
Arabidopsis thaliana is frequently grown on semisolid medium in Petri dishes, for various experiments that usually consist of two stages on two distinct growth media. Seedlings are germinated under favorable conditions followed by their transfer to another medium containing the given treatment(s). This often causes secondary effects on seedlings due to root shock, or direct and unavoidable contact of the shoot with the second medium. We have developed a simple and efficient method for the transfer of seedlings grown on semisolid medium with minimal damage. In this double-agar-layer method, seeds are germinated on a thin growth-medium-containing agar layer. Subsequently, medium blocks containing the embedded seedlings are excised and placed on the second semisolid medium supplemented with the treatment agent. Differential agar concentrations allow easy penetration of the roots into the second medium, but do not allow the shoots to come into contact with it. This unique method offers several advantages over others that are in common use, in which the seedlings are individually transferred to the second medium or alternatively grown on transfer-carrier matrices, such as filter paper, mesh and cellophane. In the presented method, the entire root system faces the growth medium, the shoots are surrounded by air at all growth stages and transfer of the seedlings is much easier. In addition, a large number of seedlings can be transferred in a single step, without stressing the plants or damaging the delicate root system. This method can also be applied to other plant species grown on semisolid media. 相似文献
11.
The Arabidopsis sku6/spiral1 gene encodes a plus end-localized microtubule-interacting protein involved in directional cell expansion 总被引:1,自引:0,他引:1
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The sku6-1 mutant of Arabidopsis thaliana exhibits altered patterns of root and organ growth. sku6 roots, etiolated hypocotyls, and leaf petioles exhibit right-handed axial twisting, and root growth on inclined agar media is strongly right skewed. The touch-dependent sku6 root skewing phenotype is suppressed by the antimicrotubule drugs propyzamide and oryzalin, and right skewing is exacerbated by cold treatment. Cloning revealed that sku6-1 is allelic to spiral1-1 (spr1-1). However, modifiers in the Columbia (Col) and Landsberg erecta (Ler) ecotype backgrounds mask noncomplementation in sku6-1 (Col)/spr1-1 (Ler) F1 plants. The SPR1 gene encodes a plant-specific 12-kD protein that is ubiquitously expressed and belongs to a six-member gene family in Arabidopsis. An SPR1:green fluorescent protein (GFP) fusion expressed in transgenic seedlings localized to microtubules within the cortical array, preprophase band, phragmoplast, and mitotic spindle. SPR1:GFP was concentrated at the growing ends of cortical microtubules and was dependent on polymer growth state; the microtubule-related fluorescence dissipated upon polymer shortening. The protein has a repeated motif at both ends, separated by a predicted rod-like domain, suggesting that it may act as an intermolecular linker. These observations suggest that SPR1 is involved in microtubule polymerization dynamics and/or guidance, which in turn influences touch-induced directional cell expansion and axial twisting. 相似文献
12.
The Arabidopsis WAVY GROWTH 2 protein modulates root bending in response to environmental stimuli 总被引:5,自引:0,他引:5
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Mochizuki S Harada A Inada S Sugimoto-Shirasu K Stacey N Wada T Ishiguro S Okada K Sakai T 《The Plant cell》2005,17(2):537-547
To understand how the direction of root growth changes in response to obstacles, light, and gravity, we characterized an Arabidopsis thaliana mutant, wavy growth 2 (wav2), whose roots show a short-pitch pattern of wavy growth on inclined agar medium. The roots of the wav2 mutant bent with larger curvature than those of the wild-type seedlings in wavy growth and in gravitropic and phototropic responses. The cell file rotations of the root epidermis of wav2-1 in the wavy growth pattern were enhanced in both right-handed and left-handed rotations. WAV2 encodes a protein belonging to the BUD EMERGENCE 46 family with a transmembrane domain at the N terminus and an alpha/beta-hydrolase domain at the C terminus. Expression analyses showed that mRNA of WAV2 was expressed strongly in adult plant roots and seedlings, especially in the root tip, the cell elongation zone, and the stele. Our results suggest that WAV2 is not involved in sensing environmental stimuli but that it negatively regulates stimulus-induced root bending through inhibition of root tip rotation. 相似文献
13.
Zornitza Andreeva Deborah Barton William J. Armour Min Y. Li Li-Fen Liao Heather L. McKellar Kylie A. Pethybridge Jan Marc 《Planta》2010,232(5):1263-1279
The phospholipase protein superfamily plays an important role in hormonal signalling and cellular responses to environmental
stimuli. There is also growing evidence for interactions between phospholipases and the cytoskeleton. In this report we used
a pharmacological approach to investigate whether inhibiting a member of the phospholipase superfamily, phospholipase C (PLC),
affects microtubules and actin microfilaments as well as root growth and morphology of Arabidopsis thaliana seedlings. Inhibiting PLC activity using the aminosteroid U73122 significantly inhibited root elongation and disrupted root
morphology in a concentration-dependent manner, with the response being saturated at 5 μM, whereas the inactive analogue U73343
was ineffective. The primary root appeared to lose growth directionality accompanied by root waving and formation of curls.
Immunolabelling of roots exposed to increasingly higher U73122 concentrations revealed that the normal transverse arrays of
cortical microtubules in the elongation zone became progressively more disorganized or depolymerized, with the disorganization
appearing within 1 h of incubation. Likewise, actin microfilament arrays also were disrupted. Inhibiting PLC using an alternative
inhibitor, neomycin, caused similar disruptions to both cytoskeletal organization and root morphology. In seedlings gravistimulated
by rotating the culture plates by 90°, both U73122 and neomycin disrupted the normal gravitropic growth of roots and etiolated
hypocotyls. The effects of PLC inhibitors are therefore consistent with the notion that, as with phospholipases A and D, PLC
likewise interacts with the cytoskeleton, alters growth morphology, and is involved in gravitropism. 相似文献
14.
Isolated barley embryos were cultivated aseptically in three different complete media. The growth of the primary root of each
embryo was measured during four days of cultivation. Embryos were cultivated on three different consistencies of media: on
agar plates, on cellulose tissue moistened with the medium and in liquid shaken cultures. The last way of cultivation yielded
the highest degree of the growth of roots. Optimum combination of conditions in shaken liquid medium was selected on the basis
of systematic study. The growth of roots of isolated embryos under appropriate conditions approaches that of roots of seedlings
cultived on moistened blotting paper. 相似文献
15.
Roots of tomato seedlings can be induced to coil by treatment with ethylene. The extent of coiling is dependent on the level of ethylene to which the seedlings are exposed and can be prevented by the incorporation of Ag ions into the growing medium. In contrast to all other tomato mutants examined, roots of the mutant diageotropica do not reorientate their growth in response to ethylene. The results of an agar penetration test indicate that roots of this mutant are agravitropic. The relationship between gravitropism and root coiling, and the origin of the ethylene modified growth pattern is discussed. 相似文献
16.
17.
MyROOT: a method and software for the semiautomatic measurement of primary root length in Arabidopsis seedlings 总被引:1,自引:0,他引:1
Isabel Betegn‐Putze Alejandro Gonzlez Xavier Sevillano David Blasco‐Escmez Ana I. Cao‐Delgado 《The Plant journal : for cell and molecular biology》2019,98(6):1145-1156
Root analysis is essential for both academic and agricultural research. Despite the great advances in root phenotyping and imaging, calculating root length is still performed manually and involves considerable amounts of labor and time. To overcome these limitations, we developed MyROOT, a software for the semiautomatic quantification of root growth of seedlings growing directly on agar plates. Our method automatically determines the scale from the image of the plate, and subsequently measures the root length of the individual plants. To this aim, MyROOT combines a bottom‐up root tracking approach with a hypocotyl detection algorithm. At the same time as providing accurate root measurements, MyROOT also significantly minimizes the user intervention required during the process. Using Arabidopsis, we tested MyROOT with seedlings from different growth stages and experimental conditions. When comparing the data obtained from this software with that of manual root measurements, we found a high correlation between both methods (R2 = 0.997). When compared with previous developed software with similar features (BRAT and EZ‐Rhizo), MyROOT offered an improved accuracy for root length measurements. Therefore, MyROOT will be of great use to the plant science community by permitting high‐throughput root length measurements while saving both labor and time. 相似文献
18.
When Arabidopsis seedlings are grown on a hard-agar plate, their primary roots show characteristic spiralling movements, apparent as waves, coils and torsions, together with a slanting toward the right-hand side. All these movements are believed to be the result of three different processes acting on the roots: circumnutation, positive gravitropism and negative thigmotropism. The basic movement of the roots is described as that of a growing right-handed helix, which, because of the root tip hitting the agar plate, is continuously switched from the right-hand to the left-hand of the growth direction, and vice versa. This movement also produces a slanting root-growth direction toward the right-hand because of the incomplete waves made by the right-handed root to the left-hand. By contrast, the torsions seen in the coils and waves are interpreted as artefacts that form as an adaptation of the three-dimensional root helix to the flat two-dimensional agar surface. 相似文献
19.
Tomato root growth, gravitropism, and lateral development: correlation with auxin transport. 总被引:13,自引:0,他引:13
Tomato (Lycopersicon esculentum, Mill.) roots were analyzed during growth on agar plates. Growth of these roots was inhibited by the auxin transport inhibitors naphthylphthalamic acid (NPA) and semicarbazone derivative I (SCB-1). The effect of auxin transport inhibitors on root gravitropism was analyzed by measurement of the angle of gravitropic curvature after the roots were reoriented 90 degrees from the vertical. NPA and SCB-1 abolished both the response of these roots to gravity and the formation of lateral roots, with SCB-1 being the more effective at inhibition. Auxins also inhibited root growth. Both auxins tested has a slight effect on the gravity response, but this effect is probably indirect, since auxins reduced the growth rate. Auxins also stimulated lateral root growth at concentration where primary root growth was inhibited. When roots were treated with both IAA and NPA simultaneously, a cumulative inhibition of root growth was found. When both compounds were applied together, analysis of gravitropism and lateral root formation indicated that the dominant effect was exerted by auxin transport inhibitors. Together, these data suggest a model for the role of auxin transport in controlling both primary and lateral root growth. 相似文献
20.
An inhibitory effect of excess moisture on the early development of Sinapis alba L. seedlings 总被引:1,自引:1,他引:0
Abstract. Mustard seedlings in the early stages of growth are sensitive to the presence of excess moisture, growth of the radicle and anthocyanin synthesis being retarded by water at a suction of less than 3 cm water for seedlings growing on a plane surface. This inhibitory effect commences at about 12-15 h from wetting of the seed. It can be counteracted by increasing the partial pressure of oxygen in the atmosphere or by de-coating the seed, suggesting that the inhibition is caused by a restricted flow of oxygen through the mucilaginous seed coat. The positive effect of gravity on anthocyanin synthesis in the developing seedling can be wholly accounted for in terms of the removal of excess moisture. This explanation largely accounts for the improved growth of the root, but a tonic effect of gravity on root growth can also be discerned. 相似文献