Seed dormancy and longevity in <Emphasis Type="Italic">Stipa tenacissima</Emphasis> L. (Poaceae)

In the present paper we studied the life history traits related to seed germination of Stipa tenacissima, a key species in semiarid environments of western Mediterranean areas. S. tenacissima is a perennial tussock grass, which has traditionally been considered to expand mainly by vegetative propagation with little or no sexual reproduction. We analysed seed longevity as well as the type of seed dormancy and the role of the seed covers from seeds collected from different populations in SE Spain. We also studied the variation in seed germinability among populations, individuals, and years and the ability of seeds of S. tenacissima to form soil seed banks. There was significant variation in seed germination among individuals, populations and years. Lemma and palea were the main factor controlling these differences since their removal promoted higher and faster germination and eliminated the differences in germination parameters among populations. However, the control of dormancy by lemma and palea was independent of their weight, suggesting that their chemical nature plays a more important role than does size in controlling seed germination. Mechanical scarification treatments (via abrasion with sand) did not affect seed germination. The decay in seed germinability two years after seed collection and the low density of viable seeds in soils one year after seed dispersal indicated that S. tenacissima forms transient soil seed banks.     

Seed bank dynamics of two obligate seeders, <Emphasis Type="Italic">Cistus monspeliensis</Emphasis> and <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis>, in relation to time since fire

Many species in Mediterranean-type ecosystems regenerate after fire by seed germination from soil seed banks. Seed bank dynamics of two of those obligate seeders, Cistus monspeliensis and Rosmarinus officinalis, were investigated in relation to stand age since fire in southwestern Portugal. Soil seed density, annual seed input, annual seed losses through germination and seed persistence were compared between species at stands differing in age since fire (5, 10 and 35 years). Soil seed density and seed input increased over the first decade after fire and were lowest at 35-year-old stands for C. monspeliensis. In R. officinalis, few seeds were produced and found in the soil at early stages, and maximum seed input and soil seed density were attained at 35-year-old stands. Soil seed density was mostly driven by seed production in both species, which is largely dependent on plant traits and population dynamics related to fire. Overall, stand age since fire had a negligible effect on seed germination, seed persistence and viability. Ten to 39% of buried seeds were not recovered after 1 year, and viability of seeds recovered was 97–100% for C. monspeliensis and only 0–3% for R. officinalis. Variation in plant traits within the seeder syndrome was evidenced by this study. R. officinalis evidenced lower seed persistence, lower proportion of viable seed produced and lower density of viable soil seed than C. monspeliensis at any stage after fire. R. officinalis is expected to depend largely on previous year seed production for population replacement after fire.     

Effects of fruit thinning on fruit and seed features of <Emphasis Type="Italic">Cistus ladanifer</Emphasis>

Fruit and seed features are the result of reproductive allocation decisions which ultimately depend on both plant availability of resources and total number of developing fruits. In this study, we manipulated fruit load in Cistus ladanifer plants by removing 0, 25 or 75% of developing fruits. Fruit features (total fruit weight, fruit-wall weight, total seed weight per fruit and seed number per fruit) were unaffected by fruit thinning, nevertheless mean seed weight increased in treated plants independently of thinning intensity. This reduced compensation was unrelated to plant size and had no consequences on fruit predation by insects. From these results it could be suggested that not only availability of resources but also morphological constraints could affect seed size in Cistus ladanifer. On the other hand, this change in seed weight could have important consequences since in this species heavy seeds perform better after fire events but light ones are advantageous in between fire recruitments.     

<Emphasis Type="Italic">CDPK</Emphasis> gene expression in somatic embryos of <Emphasis Type="Italic">Panax ginseng</Emphasis> expressing <Emphasis Type="Italic">rolC</Emphasis>

A somatic embryogenesis protocol for plant regeneration of northern red oak (Quercus rubra) was established from immature cotyledon explants. Embryogenic callus cultures were induced on Murashige and Skoog medium (MS) containing 3% sucrose, 0.24% Phytagel™, and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-d) after 4 weeks of culture in darkness. A higher response (66%) of embryogenic callus was induced on 0.45 μM 2,4-d. Higher numbers of globular- (31), heart- (17), torpedo- (12), and cotyledon-stage (8) embryos per explant were obtained by culturing embryogenic callus on MS with 3% sucrose, 0.24% Phytagel™, and devoid of growth regulators after 8 weeks culture in darkness. Continuous sub-culturing of embryogenic callus on medium containing 2,4-d yielded only compact callus. Desiccation of embryos for 3 days in darkness at 25 ± 2°C followed by cold storage at 4°C in darkness for 8 weeks favored embryo germination and development of plantlets. Cotyledon-stage embryos subjected to desiccation and chilling treatment cultured on MS with 3% sucrose, 0.24 Phytagel™, 0.44 μM 6-benzylaminopurine (BA), and 0.29 μM gibberellic acid germinated at a higher frequency (61%) than with 0.44 μM BA alone and control cultures. Germinated plantlets developed a shoot and root, were acclimatized successfully, and maintained in a growth room for plantlet development.     

Germination ecology of <Emphasis Type="Italic">Scorpiurus subvillosus</Emphasis> L. seeds: the role of temperature and storage time

Scorpiurus subvillosus L., wide spread in pastures of Mediterranean basin, is disappearing in the native pastures of the Hyblean plateau (Sicily, southern Italy), because of overgrazing and intensive management techniques. Moreover, it exhibits seed coat dormancy, which delays and reduces germination preventing its diffusion. This paper represents a first attempt in order to investigate changing in germination determined by storage time and temperature on seeds of two populations of S. subvillosus. Germination of S.␣subvillosus seeds was tested in relation to four storage time (30, 130, 200 and 360 days after harvest (DAH)), eight constant temperatures (5, 10, 15, 20, 25, 30, 35 and 40°C) and two populations of different provenience (30 and 600 m above mean sea level). The experiments were conducted either on scarified and unscarified seeds. In S. subvillosus the failure of germination under favourable conditions must be attributed␣only to seed coat, since seed scarification enhanced germination percentage with values up to 100% at almost all tested temperatures. In both treatments, but with a grater incidence in unscarified, seed germination increased gradually as temperature raised, peaking at 20–25°C, then declined with further increases of temperatures. At 40°C no germination occurred. Storage time induced a softening effect, which is somewhat limited by the natural ageing of seeds occurring from about 6 months after harvest.     

Long-term variability in seed size and seedling establishment of <Emphasis Type="Italic">Maianthemum bifolium</Emphasis>

Relatively few studies conducted in natural plants populations focus on the relationship between seed size and their germination ability and seedling establishment. Maianthemum bifolium is a perennial herb that spreads vegetatively through rhizomatous growth and reproduces through seeds. However, this species is characterized as seed and microsite limited, and under undisturbed conditions seedlings are not noted. The studies were conducted in two populations of M. bifolium in six subsequent seasons. The mean seed mass was negatively correlated both per ramet as well as in the fruit with the number of seeds, and positively with its height and the number of flowers. The long-term mean annual production of seeds in the populations was 37 and 56 seeds per m². The seeds from both populations had similarly high germination abilities that were approximately 90% under laboratory conditions, 60% in garden, and 55% in the natural habitat. Seeds from four size classes were sown and a positive correlation was noted between seedling establishment and the mass of the seeds from which they grew (r_S = 0.64). Seedling survival was also significantly correlated with seed mass.     

Seed predation limits post-fire recruitment in the waratah (<Emphasis Type="Italic">Telopea speciosissima</Emphasis>)

Seed predation may reduce recruitment in populations that are limited by the availability of seeds rather than microsites. Fires increase the availability of both seeds and microsites, but in plants that lack a soil- or canopy-stored seed bank, post-fire recruitment is often delayed compared to the majority of species. Pyrogenic flowering species, such as Telopea speciosissima, release their non-dormant seeds more than 1 year after fire, by which time seed predation and the availability of microsites may differ from that experienced by plants recruiting soon after fire. I assessed the role of post-dispersal seed predation in limiting seedling establishment after fire in T. speciosissima, in southeastern Australia. Using a seed-planting experiment, I manipulated vertebrate access to seeds and the combined cover of litter and vegetation within experimental microsites in the 2 years of natural seed fall after a fire. Losses to vertebrate and invertebrate seed predators were rapid and substantial, with 50% of seeds consumed after 2 months in exposed locations and after 5 months when vertebrates were excluded. After 7 months, only 6% of seeds or seedlings survived, even where vertebrates were excluded. Removing litter and vegetation increased the likelihood of seed predation by vertebrates, but had little influence on losses due to invertebrates. Microsites with high-density vegetation and litter cover were more likely to have seed survival or germination than microsites with low-density cover. Recruitment in pyrogenic flowering species may depend upon the release of seeds into locations where dense cover may allow them to escape from vertebrate predators. Even here, conditions suitable for germination must occur soon after seed release for seeds to escape from invertebrate predators. Seed production will also affect recruitment after any one fire, while the ability of some juvenile and most adult plants to resprout after fire buffers populations against rapid declines when there is little successful recruitment.     

Diverse bacteria isolated from root nodules of <Emphasis Type="Italic">Trifolium</Emphasis>, <Emphasis Type="Italic">Crotalaria</Emphasis> and <Emphasis Type="Italic">Mimosa</Emphasis> grown in the subtropical regions of China

We investigated the regulation of the two of the three groE operons (cpn.1 and cpn.2) of the root-nodulating bacterium R. leguminosarum strain A34. Both are heat inducible, and both have a CIRCE sequence in their upstream regions, suggesting regulation by an HrcA repressor. Mutagenesis of the CIRCE sequence upstream of cpn.1 led to an increase in the levels of cpn.1 mRNA, and knock-out of the hrcA gene increased the level of Cpn60.1 protein (the GroEL homologue encoded by the cpn.1 operon). Inactivation of the hrcA gene also caused increased expression of a 29 kDa protein that was identified as RhiA, a component of a quorum-sensing system. However, neither loss of the upstream CIRCE sequence, nor loss of HrcA function, had any effect on expression from the cpn.2 promoter. Further analysis of the cpn.2 upstream region suggested regulation could be mediated by an RpoH system, and this was confirmed by deleting the rpoH gene from the chromosome, which led to a decreased level of Cpn60.2 expression. Inactivation of RpoH led to a reduction in growth rate which could be partly compensated for by inactivation of HrcA, indicating an overlap in the in vivo function of the proteins regulated by these two systems. Accession numbers: DQ173160 (hrcA operon); DQ173161 (rpoH gene).     

Effects of the pericarp on imbibition,seed germination,and seedling establishment in seeds of <Emphasis Type="Italic">Hedysarum scoparium</Emphasis> Fisch. et Mey

Four independent experiments were designed to investigate the effects of the pericarp on seed imbibition, dehydration, germination, seedling establishment, and seed longevity in the field in seeds of Hedysarum scoparium Fisch. et Mey. The results showed that the presence of the pericarp decreased seed imbibition rates in the first 6 h, but the seeds attained significantly higher final water content after 24 h of soaking. The pericarp caused seed dormancy, and removal of the pericarp improved the germination percentage to 90 from 44%. In the pot experiment, where the level of moisture was maintained at field capacity (control), seeds with the pericarp removed had significantly improved seedling establishment. However, no statistical differences were observed in seedling establishment when the experiment was repeated under dry conditions at 40% of the field water capacity. The seedling biomass derived from seeds without the pericarp was much higher in the control but the trend was reversed under dry conditions. For seed longevity, 2 months burial in the field killed almost all seeds without the pericarp, while more than 70% of the seeds with the pericarp intact remained viable. These results indicated that the pericarp was beneficial for seedling establishment and seed longevity in arid environments. The results of this study may have practical application in grassland restoration in dry areas, especially for aerial seeding, which has been extensively used in the northern part of China.     

Heat increases germination of water-permeable seeds of obligate-seeding <Emphasis Type="Italic">Darwinia</Emphasis> species (Myrtaceae)

We examined the response of seeds to heat in four geographically restricted and one widespread species of shrubby Darwinia from the fire-prone region of southeastern Australia. These shrubs are killed by fire and rely on seed germination after a fire to maintain populations. We replicated the germination trials across several sites and several fruiting seasons for most species. Seeds had a high level of viability and were largely dispersed in a dormant state, except in D. glaucophylla, where seed dormancy varied significantly across fruiting seasons. The indehiscent fruit of all species readily imbibes moisture when wet and seeds are not considered to be ‘hard-seeded’. All species had increased seed germination in response to a limited range of heating temperatures (generally 80–100°C). Higher temperatures killed increasing proportions of seeds. This pattern was broadly consistent across species, population and seasons, although the proportion of seeds whose germination was promoted by heat varied from high (D. diminuta, D. fascicularis, D. glaucophylla) to moderate (D. biflora, D. procera). Our work highlights the importance of heat as a mechanism for influencing germination in species that are not hard-seeded. Consequently, soil temperatures during a fire should strongly influence post-fire germination levels in Darwinia. The roles of other cues that promote germination, i.e. smoke, seasonal temperatures and their interactions with heat, remain to be investigated.     

Glucose delays seed germination in<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis>

Here we report that glucose delays germination of Arabidopsis thaliana (L.) Heynh. seeds at concentrations below those known to inhibit early seedling development. This inhibition acts on embryo growth and is independent of hexokinase (HXK) function. Hormones and hormone inhibitors were applied to the germination media and several hormone biosynthesis and signalling mutants were tested on glucose media to investigate a possible role of abscisic acid (ABA), gibberellin and ethylene in the glucose-induced germination delay. Results indicate that the germination inhibition by glucose cannot be antagonized by ethylene or gibberellin and is independent of the HXK1/ABA/ABI4 signalling cascade. These findings suggest that there is a separate regulatory pathway independent of ABI2/ABI4/ABI5. Thus, in a relatively short time frame sugars utilize different signalling cascades to inhibit germination and post-germination growth, underlining the complexity of sugar responses.Abbreviations ABA Abscisic acid - ABI ABA insensitive - ACC 1-Aminocyclopropane-1-carboxylic acid - BR Brassinosteroid - CAB Chlorophyll a/b-binding protein - FUS3 Fusca3 - GA Gibberellin - GA ₃ Gibberellic acid - HXK Hexokinase - LEC1 Leafy cotyledon1 - RBCS Ribulose-1,5-bisphosphate carboxylase small subunit - WT Wild type     

Propagation of <Emphasis Type="Italic">Haemaria discolor</Emphasis> via <Emphasis Type="Italic">in vitro</Emphasis> seed germination

In vitro propagation protocol for Haemaria discolor (Ker) Lindl. var. dawsoniana by artificial cross-pollination and asymbiotic germination of seeds has been developed. Fruit set (100 %) was obtained when the pollinia and ovules of various aged flowers were used for pollination. In vitro germination of seeds obtained from capsules of various ages was achieved on half-strength Murashige and Skoog’s (MS) medium supplemented with 3 % sucrose and 0.85 % agar. The germinated seedlings were cultured on half-strength MS medium with 0.2 % activated charcoal, 8 % banana homogenate, 0.1 mg dm⁻³ 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (TDZ) and 1 mg dm⁻³ α-naphthaleneacetic acid (NAA). Ninety-six percent of plantlets survived after hardening in greenhouse.This research was supported by grant (91AS-3.1.1-CI-C3) from the Council of Agriculture, Executive Yuan of Taiwan and grants (NSC-89-2317-B055-002 and NSC-91-2317-B324-001) from the National Science Council of Taiwan. This paper is Agricultural Research Institute Contribution No. 2158.     

Gene structure and transcriptional regulation of <Emphasis Type="Italic">dnaK</Emphasis> and <Emphasis Type="Italic">dnaJ</Emphasis> genes from a psychrophilic bacterium, <Emphasis Type="Italic">Colwellia maris</Emphasis>

The dnaK and dnaJ genes, encoding heat shock proteins, were cloned from a psychrophilic bacterium, Colwellia maris. Significant homology was evident comparing DnaK and DnaJ of the psychrophilile with the counterparts of mesophilic and thermophilic bacteria. In the DnaJ protein, three conserved regions of the Hsp40 family were observed. A putative promoter similar to the sigma32 consensus sequence was found upstream of the dnaK gene. The G+C content in the 5'-untranslated region of the dnaK gene was much lower than that in the corresponding region of mesophilic bacteria. Northern-blot analysis and primer-extension analysis showed that both genes were transcribed separately as monocistronic mRNAs. Following several temperature upshifts from 10 to 26 degrees C, maximum induction of the dnaK and dnaJ mRNAs was detected at 20 degrees C, suggesting that this temperature induces the heat shock response in this bacterium. In addition, the level of the induction of the dnaJ gene was much lower than that of the dnaK gene. These findings together revealed several specific features of the heat shock response at a relatively low temperature in psychrophiles.     

Two of the three <Emphasis Type="Italic">groEL</Emphasis> homologues in <Emphasis Type="Italic">Rhizobium leguminosarum</Emphasis> are dispensable for normal growth

Although many bacteria contain only a single groE operon encoding the essential chaperones GroES and GroEL, examples of bacteria containing more than one groE operon are common. The root-nodulating bacterium Rhizobium leguminosarum contains at least three operons encoding homologues to Escherichia coli GroEL, referred to as Cpn60.1, Cpn60.2 and Cpn60.3, respectively. We report here a detailed analysis of the requirement for and relative levels of these three proteins. Cpn60.1 is present at higher levels than Cpn60.2, and Cpn60.3 protein could not be detected under any conditions although the cpn60.3 gene is transcribed under anaerobic conditions. Insertion mutations could not be constructed in cpn60.1 unless a complementing copy was present, showing that this gene is essential for growth under the conditions used here. Both cpn60.2 and cpn60.3 could be inactivated with no loss of viability, and a double cpn60.2 cpn60.3 mutant was also constructed which was fully viable. Thus only Cpn60.1 is required for growth of this organism.Dedicated to the memory of Professor V. Javier Benedí, 1957–2002     

Seed Ecology of the Invasive Tropical Tree <Emphasis Type="Italic">Parkinsonia aculeata</Emphasis>

Parkinsonia aculeata is an invasive tree native to tropical America, but introduced to Australia. Propagation and stand regeneration is mainly by seed. To gain baseline knowledge for management decisions, seed bank dynamics were monitored for two months during the fruit dispersal period at a coastal wetland in Costa Rica (native habitat), and at a coastal wetland and two semi-arid rangeland sites in Northern Queensland, Australia (introduced habitats). Seed bank densities underneath dense, uniform Parkinsonia stands were found to be lowest in the Australian wetland but highest in the Costa Rican wetland. Post-dispersal seed losses were highest in the Australian wetland, primarily due to seed germination and/or death. At the other sites, seed losses were minor during the study period, and predation was the most important cause of losses. At the two rangeland sites bruchid beetles accounted for more than 95% of the seed losses by predation. Total predation was lowest in the Costa Rican wetland. In order to test for intrinsic differences of seed characteristics, germination trials were conducted using both canopy seeds and seeds from the soil seed bank. Dormancy release and germination rate were studied under four temperature treatments. In all populations, dormancy release increased with increasing temperature, but averaged responses were significantly different between Costa Rican and Australian seed populations, and between seeds collected from the soil and from trees. Germination rate of scarified seeds was fastest at 35 °C in all tested seed populations. While high seed germination levels seem to explain low seed bank densities in the Australian wetland, the large seed banks at the rangeland sites reflect the lower incidence of favourable conditions for germination. In the Australian wetland biocontrol with bruchids is unlikely to be successful, while control by conventional methods, such as killing stands by basal bark spraying, seems feasible, due to a lower long-term risk of re-infestation from the soil seed bank. At the rangeland sites conventional control will be difficult and costly. Parkinsonia stands may be better left to their own, while bruchid populations are monitored and management efforts are concentrated on preventing further invasion.     

Mycorrhization between <Emphasis Type="Italic">Cistus ladanifer</Emphasis> L. and <Emphasis Type="Italic">Boletus edulis</Emphasis> Bull is enhanced by the mycorrhiza helper bacteria <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> Migula

Boletus edulis Bull. is one of the most economically and gastronomically valuable fungi worldwide. Sporocarp production normally occurs when symbiotically associated with a number of tree species in stands over 40 years old, but it has also been reported in 3-year-old Cistus ladanifer L. shrubs. Efforts toward the domestication of B. edulis have thus focused on successfully generating C. ladanifer seedlings associated with B. edulis under controlled conditions. Microorganisms have an important role mediating mycorrhizal symbiosis, such as some bacteria species which enhance mycorrhiza formation (mycorrhiza helper bacteria). Thus, in this study, we explored the effect that mycorrhiza helper bacteria have on the efficiency and intensity of the ectomycorrhizal symbiosis between C. ladanifer and B. edulis. The aim of this work was to optimize an in vitro protocol for the mycorrhizal synthesis of B. edulis with C. ladanifer by testing the effects of fungal culture time and coinoculation with the helper bacteria Pseudomonas fluorescens Migula. The results confirmed successful mycorrhizal synthesis between C. ladanifer and B. edulis. Coinoculation of B. edulis with P. fluorescens doubled within-plant mycorrhization levels although it did not result in an increased number of seedlings colonized with B. edulis mycorrhizae. B. edulis mycelium culture time also increased mycorrhization levels but not the presence of mycorrhizae. These findings bring us closer to controlled B. edulis sporocarp production in plantations.     

Isolation of a <Emphasis Type="Italic">Latimeria menadoensis</Emphasis><Emphasis Type="Italic">heat shock protein 70</Emphasis> (<Emphasis Type="Italic">Lmhsp70</Emphasis>) that has all the features of an inducible gene and encodes a functional molecular chaperone

Molecular chaperones facilitate the correct folding of other proteins, and heat shock proteins form one of the major classes of molecular chaperones. Heat shock protein 70 (Hsp70) has been extensively studied, and shown to be critically important for cellular protein homeostasis in almost all prokaryotic and eukaryotic systems studied to date. Since there have been very limited studies conducted on coelacanth chaperones, the main objective of this study was to genetically and biochemically characterize a coelacanth Hsp70. We have successfully isolated an Indonesian coelacanth (L. menadoensis) hsp70 gene, Lmhsp70, and found that it contained an intronless coding region and a potential upstream regulatory region. Lmhsp70 encoded a typical Hsp70 based on conserved structural and functional features, and the predicted upstream regulatory region was found to contain six potential promoter elements, and three potential heat shock elements (HSEs). The intronless nature of the coding region and the presence of HSEs suggested that Lmhsp70 was stress-inducible. Phylogenetic analyses provided further evidence that Lmhsp70 was probably inducible, and that it branched as a clade intermediate between bony fish and tetrapods. Recombinant LmHsp70 was successfully overproduced, purified and found to be functional using ATPase activity assays. Taken together, these data provide evidence for the first time that the coelacanth encodes a functional molecular chaperone system. K. W. Modisakeng and M. Jiwaji contributed equally to this study.     

A consensus map for <Emphasis Type="Italic">Cucurbita pepo</Emphasis>

Using random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), simple sequence repeats (SSR), and morphological traits, the first genetic maps for Cucurbita pepo (2n=2x=40) were constructed and compared. The two mapping populations consisted of 92 F₂ individuals each. One map was developed from a cross between an oil-seed pumpkin breeding line and a zucchini accession, into which genes for resistance to Zucchini Yellow Mosaic Virus (ZYMV) from a related species, C. moschata, had been introgressed. The other map was developed from a cross between an oil-seed pumpkin and a crookneck variety. A total of 332 and 323 markers were mapped in the two populations. Markers were distributed in each map over 21 linkage groups and covered an average of 2,200 cM of the C. pepo genome. The two maps had 62 loci in common, which enabled identification of 14 homologous linkage groups. Polyacrylamide gel analyses allowed detection of a high number of markers suitable for mapping, 10% of which were co-dominant RAPD loci. In the Pumpkin-Zucchini population, bulked segregant analysis (BSA) identified seven markers less than 7 cM distant from the locus n, affecting lignification of the seed coat. One of these markers, linked to the recessive hull-less allele (AW11-420), was also found in the Pumpkin-Crookneck population, 4 cM from n. In the Pumpkin-Zucchini population, 24 RAPD markers, previously introduced into C. pepo from C. moschata, were mapped in two linkage groups (13 and 11 markers in LGpz1 and LGpz2, respectively), together with two sequence characterized amplified region (SCAR) markers linked to genes for resistance to ZYMV.     

Overexpression of Organellar and Cytosolic <Emphasis Type="Italic">AtHSP90</Emphasis> in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Impairs Plant Tolerance to Oxidative Stress

Three AtHSP90 isoforms, cytosol-localized AtHSP90.2, chloroplast-localized AtHSP90.5, and endoplasmic reticulum (ER)-localized AtHSP90.7 genes, were constitutively overexpressed in Arabidopsis thaliana to study their functional mechanisms under oxidative stress. Overexpression of AtHSP90 genes reduced germination of transgenic seeds under oxidative stress. When exposed to 10 mM H₂O₂, AtHSP90 transgenic seedlings displayed lower activities of superoxide dismutase, catalase, and peroxidase; higher content of malondialdehyde; and higher levels of protein damage than detected in the wild type. This indicated that overexpression of AtHSP90.2, AtHSP90.5, and AtHSP90.7 in Arabidopsis impaired plant tolerance to oxidative stress. Moreover, overexpression of chloroplast- and ER-localized AtHSP90 resulted in lower resistance to oxidative stress than that of cytosolic AtHSP90. This suggested that HSP90.2, HSP90.5, and HSP90.7 localized in different cellular compartments were involved in different functional mechanisms during oxidative stress.