共查询到20条相似文献,搜索用时 31 毫秒
1.
2.
Expansins are cell wall proteins characterised by their ability to stimulate wall loosening during cell expansion. The expression of some expansin isoforms is clearly correlated with growth and the external application of expansins can stimulate cell expansion in vivo in several systems. We report here the expression of a heterologous expansin coding sequence in transgenic tomato plants (Lycopersicon esculentum Mill.) under the control of a constitutive promoter. In some transgenic lines with high levels of expansin activity extractable from cell walls, we observed alterations of growth: mature plants were stunted, with shorter leaves and internodes, and dark-grown seedlings had shorter and wider hypocotyls than their wild-type counterparts. Examination of hypocotyl sections revealed similar differences at the cellular level: cortical and epidermal cells were shorter and wider than those from wild-type seedlings. The observed stimulation of radial expansion did not compensate for the decreased elongation, and overall growth was reduced in the transgenics. As this observation can seem paradoxical given the known effect of expansins on isolated cell walls, we examined the mechanical behaviour of transgenic tissue. We measured a decrease in hypocotyl elongation in response to acidic pH in the transformants. This result may account for the alterations in cell expansion, and could itself be explained by a reduced susceptibility of transgenic cell walls to expansin action. 相似文献
3.
Ectopic expression of a wood-abundant expansin PttEXPA1 promotes cell expansion in primary and secondary tissues in aspen 总被引:1,自引:0,他引:1
Gray-Mitsumune M Blomquist K McQueen-Mason S Teeri TT Sundberg B Mellerowicz EJ 《Plant biotechnology journal》2008,6(1):62-72
Expansins are primary agents inducing cell wall extension, and are therefore obvious targets in biotechnological applications aimed at the modification of cell size in plants. In trees, increased fibre length is a goal of both breeding and genetic engineering programmes. We used an α-expansin Ptt EXPA1 that is highly abundant in the wood-forming tissues of hybrid aspen ( Populus tremula L. × P. tremuloides Michx.) to evaluate its role in fibre elongation and wood cell development. Ptt EXPA1 belongs to Subfamily A of α-expansins that have conserved motifs at the N- and C-termini of the mature protein. When PttEXPA1 was over-expressed in aspen, an extract of the cell wall-bound proteins of the transgenic plants exhibited an increased expansin activity on cellulose–xyloglucan composites in vitro , indicating that Ptt EXPA1 is an active expansin. The transgenic lines exhibited increased stem internode elongation and leaf expansion, and larger cell sizes in the leaf epidermis, indicating that Ptt EXPA1 protein is capable of increasing the growth of these organs by enhancing cell wall expansion in planta . Wood cell development was also modified in the transgenic lines, but the effects were different for vessel elements and fibres, the two main cell types of aspen wood. Ptt EXPA1 stimulated fibre, but not vessel element, diameter growth, and marginally increased vessel element length, but did not affect fibre length. The observed differences in responsiveness to expansin of these cell types are discussed in the light of differences in their growth strategies and cell wall composition. 相似文献
4.
5.
Detection of expansin proteins and activity during tomato fruit ontogeny 总被引:19,自引:0,他引:19
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Expansins are plant proteins that have the capacity to induce extension in isolated cell walls and are thought to mediate pH-dependent cell expansion. J.K.C. Rose, H.H. Lee, and A.B. Bennett ([1997] Proc Natl Acad Sci USA 94: 5955-5960) reported the identification of an expansin gene (LeExp1) that is specifically expressed in ripening tomato (Lycopersicon esculentum) fruit where cell wall disassembly, but not cell expansion, is prominent. Expansin expression during fruit ontogeny was examined using antibodies raised to recombinant LeExp1 or a cell elongation-related expansin from cucumber (CsExp1). The LeExp1 antiserum detected expansins in extracts from ripe, but not preripe tomato fruit, in agreement with the pattern of LeExp1 mRNA accumulation. In contrast, antibodies to CsExp1 cross-reacted with expansins in early fruit development and the onset of ripening, but not at a later ripening stage. These data suggest that ripening-related and expansion-related expansin proteins have distinct antigenic epitopes despite overall high sequence identity. Expansin proteins were detected in a range of fruit species and showed considerable variation in abundance; however, appreciable levels of expansin were not present in fruit of the rin or Nr tomato mutants that exhibit delayed and reduced softening. LeExp1 protein accumulation was ethylene-regulated and matched the previously described expression of mRNA, suggesting that expression is not regulated at the level of translation. We report the first detection of expansin activity in several stages of fruit development and while characteristic creep activity was detected in young and developing tomato fruit and in ripe pear, avocado, and pepper, creep activity in ripe tomato showed qualitative differences, suggesting both hydrolytic and expansin activities. 相似文献
6.
7.
Comparative analysis of expansin gene codon usage patterns among eight plant species 总被引:1,自引:0,他引:1
Hao Zhang Jian Li Ruixue Wang Junkai Zhi Peng Yin 《Journal of biomolecular structure & dynamics》2019,37(4):910-917
Expansins are essential components of plant cell wall and play an important role in plant growth, development, and stress resistance via loosening function. To understand the codon usage pattern of expansin genes, we gained the sequence data of expansin genes from eight plant species. Statistics analysis showed obvious codon characteristics between monocot and dicot plants. Comparably, expansin genes in monocot plants had really higher GC content, more high-frequency codons, and more optimal codons than that in dicot plants. Several monocot plants performed somehow as dicot plants in a few characters. Codon information of expansin genes might contribute to the understanding of the relationship and evolution clues between monocot and dicot plants. It further gained insight into the improvement of the gene expression and roles. 相似文献
8.
Ermel FF Follet-Gueye ML Cibert C Vian B Morvan C Catesson AM Goldberg R 《Planta》2000,210(5):732-740
The development of pectin structural features during the differentiation of cambial derivatives was investigated in aspen
(Populus tremula L. × P. tremuloides Michx.) using biochemical and immunocytochemical methods. Comparisons were also made between active and resting tissues.
Active tissues, in particular cambial cells and phloem derivatives, were characterized by a high pectin content. Use of antibodies
raised against arabinan side chains of rhamnogalacturonan 1 (LM6), as well as biochemical analysis, revealed an obvious decrease
from the cortex to the differentiating xylem. Galactan side chains, detected with LM5 antibodies, were present mainly in the
cambial zone and enlarging xylem cells. In contrast, they were totally absent from sieve-tube cell walls. Image analysis of
LM5 immunogold labelling in the cambial zone showed a clustered distribution of galactan epitopes in the radial walls, a distribution
which might result from the association of two different periodic processes, namely the exocytosis of galactan and wall expansion.
Cessation of cambial activity was characterized by cell wall thickening accompanied by a sharp decrease in the relative amount
of pectin and a lowering of the degree of methylesterification. The data provide evidence that the walls of phloem and xylem
cells differ in their pectin composition even at a very early stage of commitment. These differences offer useful tools for
identifying the initial cells among their immediate neighbours.
Received: 12 June 1999 / Accepted: 20 October 1999 相似文献
9.
It is desirable that the expression of transgenes in genetically modified crops is restricted to the tissues requiring the
encoded activity. To this end, we have studied the ability of the heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase
(Rubisco) small-subunit (SSU) gene promoters, RBCS3CP (0.8 kbp) from tomato (hycopersion esculentum Mill.) and SRS1P (1.5 kbp) from soybean (Glycine max [h.] Mers.), to drive expression of the β-glucuronidase (gusA) marker gene in apple (Malus pumila Mill.). Transgenic lines of cultivar Greensleeves were produced by Agrobacterium-mediated transformation and the level of gusA expression in the vegetative tissues of young plants was compared with that produced using the cauliflower mosaic virus (CaMV)
35S promoter. These quantitative GUS data were assessed for their relationship to the copy number of transgene loci. The precise
location of GUS activity in leaves was identified histochemically. The heterologous SSU promoters were active primarily in
the green vegetative tissues of apple, although activity in the roots was noticeably higher with the RBCS3C promoter than with the SRS1 promoter. The mean GUS activity in leaf tissue of the SSU promoter transgenics was approximately half that of plants containing
the CaMV 35S promoter. Histochemical analysis demonstrated that GUS activity was localised to the mesophyll and palisade cells
of the leaf. The influence of light on expression was also determined. The activity of the SRS1 promoter was strictly dependent on light, whereas that of the RBCS3C promoter appeared not to be. Both SSU promoters would be suitable for the expression of transgenes in green photosynthetic
tissues of apple.
Received: 15 June 1999 / Accepted: 12 August 1999 相似文献
10.
Immunolocalization of LeAGP-1, a modular arabinogalactan-protein, reveals its developmentally regulated expression in tomato 总被引:1,自引:0,他引:1
Arabinogalactan-proteins (AGPs) are highly glycosylated cell surface proteins that are thought to function in plant growth
and development. The developmentally regulated expression of LeAGP-1, a novel and major AGP in tomato, was examined in different
organs and tissues of tomato (Lycopersicon esculentum Mill. cv. UC82B) plants with an anti-peptide antibody (i.e. the PAP antibody) directed specifically against the lysine-rich
subdomain of the LeAGP-1 core protein. During cell differentiation in tomato plants, LeAGP-1 was associated with cell wall
thickening and lignification of particular cell types. Specifically, LeAGP-1 was detected in secondary wall thickenings of
maturing metaxylem and secondary xylem tracheary elements in roots and stems, and in thickened cell walls of phloem sieve
elements. However, LeAGP-1 was also present in thin-walled, cortical parenchyma cells of seedling roots as well as thick-walled
collenchyma cells in young stems, both of which are not lignified. Based on these observed patterns, possible roles for LeAGP-1
in plant growth and development are discussed.
Received: 17 August 1999 / Accepted: 7 October 1999 相似文献
11.
Rice is the only cereal in which germination and coleoptile elongation occur in hypoxia or anoxia. Little is known of the
molecular basis directly underlying coleoptile cell extension. In this paper, we describe the expression of α-expansin genes
in embryos during seed development and young seedlings grown under various oxygen concentrations. The genes Os-EXP2 and Os-EXP1 were predominantly expressed in the developing seeds, mainly in newly developed leaves, coleoptiles, and seminal roots. These
expansins expressed in the developing seeds may give cells the potential to expand after seed imbibition begins. In coleoptiles,
Os-EXP4 and Os-EXP2 mRNAs were greatly induced by submergence, while they were weakly detected in aerobic or anoxic conditions. Under submerged
soil conditions, the signals hybridized with probes Os-EXP4 and Os-EXP2 in coleoptiles were strongest when coleoptiles elongated in the water layer. These data show that expansin gene expression
is highly correlated with coleoptile elongation in response to oxygen concentrations. The Os-EXP4 gene was also expressed in leaves, mesocotyls, and coleorhizas of young seedlings. The growth of these tissues was also correlated
with the presence of expansins. Therefore, the evidence derived from this study clearly demonstrates that expansins are indispensable
for the growing tissues of rice seedlings.
Received: 23 December 1999 / Accepted: 24 February 2000 相似文献
12.
Expression of a Petunia inflata pectin methyl esterase in Solanum tuberosum L. enhances stem elongation and modifies cation distribution 总被引:2,自引:0,他引:2
Transgenic potato (Solanum tuberosum L.) plants were constructed with a Petunia inflata-derived cDNA encoding a pectin methyl esterase (PME; EC 3.1.1.11) in sense orientation under the control of the cauliflower
mosaic virus 35S promoter. The PME activity was elevated in leaves and tubers of the transgenic lines but slightly reduced
in apical segments of stems from mature plants. Stem segments from the base of juvenile PME-overexpressing plants did not
differ in PME activity from the control, whereas in apical parts PME was less active than in the wild-type. During the early
stages of development stems of these trangenic plants elongated more rapidly than those of the wild-type. Further evidence
that overexpression of a plant-derived PME has an impact on plant development is based on modifications of tuber yield, which
was reduced in the transgenic lines. Cell walls from transgenic tubers showed significant differences in their cation-binding
properties in comparison with the wild-type. In particular, cell walls displayed increased affinity for sodium and calcium,
while potassium binding was constant. Furthermore, the total ion content of transgenic potatoes was modified. Indications
of PME-mediated differences in the distribution of ions in transgenic plants were also obtained by monitoring relaxations
of the membrane potential of roots subsequent to changes in the ionic composition of the bathing solution. However, no effects
on the chemical structure of pectin from tuber cell walls could be detected.
Received: 24 March 1999 / Accepted: 20 August 1999 相似文献
13.
Tomato (Lycopersicon esculentum Mill.) prosystemin in fusion with a viral signal peptide was expressed in Sf21 insect cell cultures after infection with recombinant baculoviruses. Prosystemin was purified from culture supernatants
and its identity was confirmed by N-terminal sequence and mass-spectral analyses. Recombinant prosystemin was found to be
equally active as compared to systemin in inducing the expression of wound-response genes in tomato plants. In cultured cells
of L. peruvianum, prosystemin elicited a rapid alkalinization of the growth medium. The timing and dose-dependence of the alkalinization response
were found to be identical for prosystemin and systemin, respectively. Prosystemin-triggered defense responses were inhibited
by a competitive antagonist of systemin activity, indicating that the systemin sequence within the primary structure of prosystemin
determines its activity.
Received: 30 August 1999 / Accepted: 6 December 1999 相似文献
14.
The study of Carbohydrate-Active enZymes (CAZymes) associated with plant cell wall metabolism is important for elucidating the developmental mechanisms of plants and also for the utilization of plants as a biomass resource. The use of recombinant proteins is common in this context, but heterologous expression of plant proteins is particularly difficult, in part because the presence of many cysteine residues promotes denaturation, aggregation and/or protein misfolding. In this study, we evaluated two phenotypes of methylotrophic yeast Pichia pastoris as expression hosts for expansin from peach (Prunus persica (L.) Batsch, PpEXP1), which is one of the most challenging targets for heterologous expression. cDNAs encoding wild-type expansin (PpEXP1_WT) and a mutant in which all cysteine residues were replaced with serine (PpEXP1_CS) were each inserted into expression vectors, and the protein expression levels were compared. The total amount of secreted protein in PpEXP1_WT culture was approximately twice that of PpEXP1_CS. However, the amounts of recombinant expansin were 0.58 and 4.3 mg l−1, corresponding to 0.18% and 2.37% of total expressed protein, respectively. This 13-fold increase in production of the mutant in P. pastoris indicates that the replacement of cysteine residues stabilizes recombinant PpEXP1. 相似文献
15.
Expansins are unique plant cell wall proteins that possess the ability to induce immediately cell wall extension in vitro and cell expansion in vivo. To investigate the biological functions of expansins that are abundant in wood-forming tissues, we cloned two expansin genes from the differentiating xylem of Chinese fir (Cunninghamia lanceolata (Lamb.) Hook). Phylogenetic reconstruction indicated that they belong to α-expansin (EXPA), named ClEXPA1 and ClEXPA2. Expression pattern analysis demonstrated that they are preferentially expressed in the cambium region. Overexpression of ClEXPA1 and ClEXPA2 in tobacco plants yielded pleiotropic phenotypes of plant height, stem diameter, leaf number and seed pod. The height and diameter growth of the 35S(pro) :ClEXPA1 and 35S(pro) :ClEXPA2 transgenic plants were increased drastically, exhibiting an enlargement of pith parenchyma cell size. Isolated cell walls of ClEXPA1 and ClEXPA2 overexpressors contained 30%-50% higher cellulose contents than the wild type, accompanied by a thickening of the cell walls in the xylem region. Both ClEXPA1 and ClEXPA2 are involved in plant growth and development, with a partially functional overlap. Expansins are not only able to induce cell expansion in different tissues/organs in vivo, but they also can act as a potential activator during secondary wall formation by directly or indirectly affecting cellulose metabolism, probably in a cell type-dependent manner. 相似文献
16.
17.
18.
Role of extensin peroxidase in tomato (Lycopersicon esculentum Mill.) seedling growth 总被引:3,自引:0,他引:3
It is proposed that inhibition of extensin peroxidase activity leads to a less rigid cell wall and thus promotes cell expansion
and plant growth. A low-molecular-weight inhibitor derived from the cell walls of suspension-cultured tomato cells was found
to completely inhibit extensin peroxidase-mediated extensin cross-linking in vitro at a concentration of 260 μg/ml. The inhibitor
had no effect upon guaiacol oxidation catalyzed by extensin peroxidase or horseradish peroxidase. We have demonstrated that
the light-irradiated inhibition of plant growth may be partially offset by inhibition of endogenous extensin peroxidase activity.
Overall plant growth was enhanced by up to 15% in the presence of inhibitor relative to control plants. Inhibitor-treated
and illuminated tomato hypocotyls grew up to 15% taller than untreated controls. The inhibitor had no effect upon etiolated
plants over a 15-d period, suggesting that only low levels of peroxidase-mediated cross-linking can be found in the cell walls
of etiolated plants. SDS-PAGE/Western blots of ionically bound protein from both etiolated and illuminated hypocotyls identified
a doublet at 57/58.5 kDa which is immuno-reactive with antibodies raised to tomato extensin peroxidase. Levels of the 58.5-kDa
protein, determined by SDS-PAGE, were at least threefold higher in illuminated tomato hypocotyls than in etiolated hypocotyls.
Three fold higher levels of extensin peroxidase, elevated in-vitro extensin cross-linking activity and 15% higher levels of
cross-linked, non-extractable extensin were observed in illuminated tomato hypocotyls compared with etiolated tomato hypocotyls.
This suggests that white-light inhibition of tomato hypocotyl growth appears to be mediated, at least partially, by deposition
of cell wall extensin, a process regulated by Mr-58,500 extensin peroxidase. Our results indicate that the contribution of peroxidase-mediated extensin deposition to plant
cell wall architecture may have an important role in plant growth.
Received: 22 July 1999 / Accepted: 11 October 1999 相似文献
19.
Expression of six expansin genes in relation to extension activity in developing strawberry fruit. 总被引:18,自引:0,他引:18
Expansins are proteins which have been demonstrated to induce cell wall extension in vitro. The identification and characterization of six expansin cDNAs from strawberry fruit, termed FaExp3 to FaExp7, as well as the previously identified FaExp2 is reported here. Analysis of expansin mRNAs during fruit development and in leaves, roots and stolons revealed a unique pattern of expression for each cDNA. FaExp3 mRNA was present at much lower levels than the other expansin mRNAs and was expressed in small green fruit and in ripe fruit. FaExp4 mRNA was present throughout fruit development, but was more strongly expressed during ripening. FaExp5 was the only clone to show fruit specific expression which was up-regulated at the onset of ripening. FaExp6 and FaExp7 mRNAs were present at low levels in the fruit with highest expression in stolon tissue. During fruit development FaExp6 had the highest expression at the white, turning and orange stages whereas expression of FaExp7 was highest in white fruit. The expression profiles of FaExp2 and FaExp5 in developing fruit were similar except that FaExp2 was induced at an earlier stage. Analysis of expansin protein by Western blotting using an antibody raised against CsExp1 from cucumber hypocotyls identified two bands of 29 and 31 kDa from developing fruit. Protein extracts from developing fruit were assayed for extension activity. Considerable rates of extension were observed with extracts from ripening fruit, but no extension was observed with protein from unripe green fruit. These results demonstrate the presence of at least six expansin genes in strawberry fruit and that during ripening the fruit acquires the ability to cause extension in vitro, characteristic of expansin action. 相似文献
20.
The temporal and spatial patterns of storage-globulin mobilization were immunohistochemically pursued in the embryonic axis
and cotyledons of vetch seed (Vicia sativa L.) during germination and early seedling growth. Embryonic axes as well as cotyledons of mature seeds contain protein bodies
with stored globulins. Prevascular strands of axes and cotyledons, the radicle and epidermal layers of axis organs were nearly
exclusively stained by vicilin antibodies whereas the cotyledonous storage mesophyll gave similar staining for vicilin and
legumin. Globulin breakdown started locally where growth and differentiation commenced in the axis. There, vicilin mobilization
preceded legumin mobilization. Thus vicilin represents the initial source of amino acids for early growth and differentiation
processes in vetch. Legumin presumably only serves as a bulk amino acid source for subsequent seedling growth during postgerminative
globulin degradation. During the first 2–3 d after the start of imbibition the axis was depleted of globulins whereas no decrease
in immunostainability was detected in the cotyledons except in their vascular strands where immunostainability was almost
completely lost at this time. Continuous vascular strands were established at the third day when globulin breakdown was finished
in the axis but had just started in the cotyledon mesophyll. Protein mobilization proceeded in a small zone from the epidermis
towards the vascular strands in the center of the cotyledons. In this zone the storage cells, which initially appeared densely
packed with starch grains and protein bodies, concomitantly transformed into cells with a large central vacuole and only a
thin cytoplasmic layer attached to the cell wall. These results agree well with the hypothesis that during the first 2 d after
imbibition the axis is autonomous in amino acid provision. After the endogenous reserves of the axis are depleted and the
conductive tissue has differentiated, globulins are mobilized in the cotyledons, suggesting that then the amino acid supply
is taken over by the cotyledons. For comparison with other degradation patterns we used garden bean (Phaseolus vulgaris L) and rape (Brassica napus L.) as reference plants.
Received: 3 August 1999 / Accepted: 11 December 1999 相似文献