Showing papers on "Germination published in 2007"

Identification of Two Protein Kinases Required for Abscisic Acid Regulation of Seed Germination, Root Growth, and Gene Expression in Arabidopsis

Abstract: Abscisic acid (ABA) is an important phytohormone regulating various plant processes, including seed germination. Although phosphorylation has been suggested to be important, the protein kinases required for ABA signaling during seed germination and seedling growth remain elusive. Here, we show that two protein kinases, SNF1-RELATED PROTEIN KINASE2.2 (SnRK2.2) and SnRK2.3, control responses to ABA in seed germination, dormancy, and seedling growth in Arabidopsis thaliana. A snrk2.2 snrk2.3 double mutant, but not snrk2.2 or snrk2.3 single mutants, showed strong ABA-insensitive phenotypes in seed germination and root growth inhibition. Changes in seed dormancy and ABA-induced Pro accumulation consistent with ABA insensitivity were also observed. The snrk2.2 snrk2.3 double mutant had a greatly reduced level of a 42-kD kinase activity capable of phosphorylating peptides from ABF (for ABA Response Element Binding Factor) transcription factors. ABA-induced expression of several genes whose promoters contain an ABA response element (ABRE) was reduced in snrk2.2 snrk2.3, suggesting that the mechanism of SnRK2.2 and SnRK2.3 action in ABA signaling involves the activation of ABRE-driven gene expression through the phosphorylation of ABFs. Together, these results demonstrate that SnRK2.2 and SnRK2.3 are redundant but key protein kinases that mediate a major part of ABA signaling in Arabidopsis.

Repression of AUXIN RESPONSE FACTOR10 by microRNA160 is critical for seed germination and post-germination stages.

Abstract: AUXIN RESPONSE FACTORS (ARFs) are transcription factors involved in auxin signal transduction during many stages of plant growth development. ARF10, ARF16 and ARF17 are targeted by microRNA160 (miR160) in Arabidopsis thaliana. Here, we show that negative regulation of ARF10 by miR160 plays important roles in seed germination and post-germination. Transgenic plants expressing an miR160-resistant form of ARF10, which has silent mutations in the miRNA target site (termed mARF10), exhibited developmental defects such as serrated leaves, curled stems, contorted flowers and twisted siliques. These phenotypes were not observed in wild-type plants or plants transformed with the targeted ARF10 gene. During sensu stricto germination and post-germination, mARF10 mutant seeds and plants were hypersensitive to ABA in a dose-dependent manner. ABA hypersensitivity was mimicked in wild-type plants by exogenous auxin. In contrast, overexpression of MIR160 (35S:MIR160) resulted in reduced sensitivity to ABA during germination. Transcriptome analysis of germinating ARF10 and mARF10 seeds indicated that typical ABA-responsive genes expressed during seed maturation were overexpressed in germinating mARF10 seeds. These results indicate that negative regulation of ARF10 by miR160 plays a critical role in seed germination and post-embryonic developmental programs, at least in part by mechanisms involving interactions between ARF10-dependent auxin and ABA pathways.

ROS production and protein oxidation as a novel mechanism for seed dormancy alleviation.

Abstract: *Summary At harvest, sunflower (Helianthus annuus L.) seeds are dormant and unable to germinate at temperatures below 15 � C. Seed storage in the dry state, known as after-ripening, is associated with an alleviation of embryonic dormancy allowing subsequent germination at suboptimal temperatures. To identify the process by which dormancy is broken during after-ripening, we focused on the role of reactive oxygen species (ROS) in this phenomenon. After-ripening entailed a progressive accumulation of ROS, namely superoxide anions and hydrogen peroxide, in cells of embryonic axes. This accumulation, which was investigated at the cellular level by electron microscopy, occurred concomitantly with lipid peroxidation and oxidation (carbonylation) of specific embryo proteins. Incubation of dormant seeds for 3 h in the presence of hydrogen cyanide (a compound that breaks dormancy) or methylviologen (a ROS-generating compound) also released dormancy and caused the oxidation of a specific set of embryo proteins. From these observations, we propose a novel mechanism for seed dormancy alleviation. This mechanism involves ROS production and targeted changes in protein carbonylation patterns.

Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana.

Abstract: Summary Despite much effort, a robust protocol for in vitro germination of Arabidopsis thaliana pollen has been elusive. Here we show that controlled temperatures, a largely disregarded factor in previous studies, and a simple optimized medium, solidified or liquid, yielded pollen germination rates above 80% and pollen tube lengths of hundreds of microns, with both Columbia and Landsberg erecta (Ler) ecotypes. We found that pollen germination and tube growth were dependent on pollen density in both liquid and solid medium. Pollen germination rates were not substantially affected by flower or plant age. The quartet1 mutation negatively affected pollen germination, especially in the Ler ecotype. This protocol will facilitate functional analyses of insertional mutants affecting male gametophyte function, and should allow detailed gene expression analyses during pollen tube growth. Arabidopsis thaliana can now be included on the list of plant species that are suitable models for physiological studies of pollen tube elongation and tip growth.

The Arabidopsis Aleurone Layer Responds to Nitric Oxide, Gibberellin, and Abscisic Acid and Is Sufficient and Necessary for Seed Dormancy

Abstract: Seed dormancy is a common phase of the plant life cycle, and several parts of the seed can contribute to dormancy. Whole seeds, seeds lacking the testa, embryos, and isolated aleurone layers of Arabidopsis (Arabidopsis thaliana) were used in experiments designed to identify components of the Arabidopsis seed that contribute to seed dormancy and to learn more about how dormancy and germination are regulated in this species. The aleurone layer was found to be the primary determinant of seed dormancy. Embryos from dormant seeds, however, had a lesser growth potential than those from nondormant seeds. Arabidopsis aleurone cells were examined by light and electron microscopy, and cell ultrastructure was similar to that of cereal aleurone cells. Arabidopsis aleurone cells responded to nitric oxide (NO), gibberellin (GA), and abscisic acid, with NO being upstream of GA in a signaling pathway that leads to vacuolation of protein storage vacuoles and abscisic acid inhibiting vacuolation. Molecular changes that occurred in embryos and aleurone layers prior to germination were measured, and these data show that both the aleurone layer and the embryo expressed the NO-associated gene AtNOS1, but only the embryo expressed genes for the GA biosynthetic enzyme GA3 oxidase.

Seed dormancy release in Arabidopsis Cvi by dry after‐ripening, low temperature, nitrate and light shows common quantitative patterns of gene expression directed by environmentally specific sensing

Abstract: The depth of seed dormancy can be influenced by a number of different environmental signals, but whether a common mechanism underlies this apparently similar response has yet to be investigated. Full-genome microarrays were used for a global transcript analysis of Arabidopsis thaliana Cape Verde Island accession seeds exposed to dry after-ripening (AR), or low temperature, nitrate and light when imbibed. Germination studies showed that the sensitivity of imbibed seeds to low temperature, nitrate and light was dependant upon the length of time spent AR following harvest. Seeds had an absolute requirement for light to complete dormancy release in all conditions, but this effect required an exposure to a prior dormancy relieving environment. Principal component analyses of the expression patterns observed grouped physiological states in a way that related to the depth of seed dormancy, rather than the type of environmental exposure. Furthermore, opposite changes in transcript abundance of genes in sets associated with dormancy, or dormancy relief through AR, were also related to the depth of dormancy and common to different environments. Besides these common quantitative changes, environment-specific gene expression patterns during dormancy relief are also described. For example, higher transcript abundance for genes linked to the process of nitrate accumulation, and nitrate reduction was associated with dormancy relief. The quantity of GA3ox1 transcripts increased during dormancy relief in all conditions, in particular when dormancy relief was completed by exposure to light. This contrasts with transcripts linked to abscisic acid (ABA) synthesis, which declined. The results are consistent with a role for the ABA/gibberellic acid balance in integrating dormancy-relieving environmental signals.

Proteomic analysis of rice (Oryza sativa) seeds during germination

Abstract: Although seed germination is a major subject in plant physiological research, there is still a long way to go to elucidate the mechanism of seed germination. Recently, functional genomic strategies have been applied to study the germination of plant seeds. Here, we conducted a proteomic analysis of seed germination in rice (Oryza sativa indica cv. 9311) - a model monocot. Comparison of 2-DE maps showed that there were 148 proteins displayed differently in the germination process of rice seeds. Among the changed proteins, 63 were down-regulated, 69 were up-regulated (including 20 induced proteins). The down-regulated proteins were mainly storage proteins, such as globulin and glutelin, and proteins associated with seed maturation, such as "early embryogenesis protein" and "late embryogenesis abundant protein", and proteins related to desiccation, such as "abscisic acid-induced protein" and "cold-regulated protein". The degradation of storage proteins mainly happened at the late stage of germination phase II (48 h imbibition), while that of seed maturation and desiccation associated proteins occurred at the early stage of phase II (24 h imbibition). In addition to alpha-amylase, the up-regulated proteins were mainly those involved in glycolysis such as UDP-glucose dehydrogenase, fructokinase, phosphoglucomutase, and pyruvate decarboxylase. The results reflected the possible biochemical and physiological processes of germination of rice seeds.

ABA-Hypersensitive Germination1 encodes a protein phosphatase 2C, an essential component of abscisic acid signaling in Arabidopsis seed.

Abstract: The phytohormone abscisic acid (ABA) regulates physiologically important stress and developmental responses in plants. To reveal the mechanism of response to ABA, we isolated several novel ABA-hypersensitive Arabidopsis thaliana mutants, named ahg (ABA-hypersensitive germination). ahg1-1 mutants showed hypersensitivity to ABA, NaCl, KCl, mannitol, glucose and sucrose during germination and post-germination growth, but did not display any significant phenotypes in adult plants. ahg1-1 seeds accumulated slightly more ABA before stratification and showed increased seed dormancy. Map-based cloning of AHG1 revealed that ahg1-1 has a nonsense mutation in a gene encoding a novel protein phosphatase 2C (PP2C). We previously showed that the ahg3-1 mutant has a point mutation in the AtPP2CA gene, which encodes another PP2C that has a major role in the ABA response in seeds (Yoshida et al., 2006b). The levels of AHG1 mRNA were higher in dry seeds and increased during late seed maturation--an expression pattern similar to that of ABI5. Transcriptome analysis revealed that, in ABA-treated germinating seeds, many seed-specific genes and ABA-inducible genes were highly expressed in ahg1-1 and ahg3-1 mutants compared with the wild-type. Detailed analysis suggested differences between the functions of AHG1 and AHG3. Dozens of genes were expressed more strongly in the ahg1-1 mutant than in ahg3-1. Promoter-GUS analyses demonstrated both overlapping and distinct expression patterns in seed. In addition, the ahg1-1 ahg3-1 double mutant was more hypersensitive than either monogenic mutant. These results suggest that AHG1 has specific functions in seed development and germination, shared partly with AHG3.

A review on the role of endozoochory in seed germination.

Abstract: A large fraction of living plants produce fruits that attract animals (64% and 27% of gymnosperm and angiosperm lineages, respectively; Herrera and Pellmyr, 2002). In consuming them, animals can spread the seeds to more or less distant sites from the parent plant, thus contributing to plant regeneration and colonization of new sites (Willson and Traveset, 2000, and references therein). For millions of years both the pulp and seeds of fruit have been subjected to selective pressures exerted by frugivores and granivores. Fruit consumers, specifically, may show preferences for fruit traits such as size, shape, chemical composition and others, and have specific morphologies and physiologies of the digestive tract that affect the survival probability of the ingested seeds in different ways (Herrera and Pellmyr, 2002, and references therein). Seed traits are under further selective pressures imposed by a number of biotic (antagonistic fungi, insects and microbes; Cipollini and Levey, 1997) and abiotic factors (e.g. light, temperature, rainfall; Holl, 1999; Leishman et al., 2000) that influence the dormancy period, germination time and/or future seedling growth, which can ultimately determine germination and seedling success (Verdú and Traveset, 2005, and references therein). Therefore, the specific seed traits in an endozoochorous plant are a result of the combined selection imposed by frugivores, granivores and other biotic and abiotic factors. In this chapter, we evaluate the role of endozoochory on seed germination. We review the information gathered on germination patterns in experiments aimed at examining the effect of a seed’s passage through a frugivore’s gut, and give further directions on methods for future studies. Fruit treatment in the disperser’s digestive tract (which includes pulp separation from seeds and treatment of the seeds) can determine the capacity of seeds to germinate and, thus, is one of the components of

Seed dormancy and germination stimulation syndromes for Australian temperate species

Abstract: Understanding seed germination biology and the dynamics of seed dormancy is essential to developing reliable germination techniques. This paper presents some new data and reviews recent findings from germination studies on Australian species, with respect to the role of temperature and moisture in the control of dormancy and germination. A seed-burial experiment was conducted over a 1-year period (January–December) with seeds of Acanthocarpus preissii Lehm., Astroloma xerophyllum (DC.) Sond., Gahnia grandis (Labill.) S.T.Blake, Goodenia scaevolina F.Muell. and Tersonia cyathiflora (Fenzl) J.W.Green to determine the fluctuations in temperature and moisture seeds naturally experience in the buried environment. All seeds became hydrated during autumn (March) while soil temperatures were >15°C, suggestive of a period of warm stratification before the onset of cooler winter temperatures appropriate for germination. Evidence of rapid wetting and drying of seeds in the soil environment was also present. Laboratory experiments testing stratification as a means for dormancy loss showed that several weeks of warm stratification at 26/13°C or 33/18°C promoted germination of Lomandra preissii (Endl.) Ewart, Marianthus bicolor (Putt.) F.Muell. and Xyris lanata R.Br. seeds. X. lanata seeds also responded to several weeks of cold stratification at 5°C. By integrating this new data with other published data on germination of Australian species, diagrammatic models of germination timing, dormancy syndromes and propagation strategies for temperate Australian species are presented as working hypotheses to help direct future research.

Seed after-ripening is a discrete developmental pathway associated with specific gene networks in Arabidopsis.

Abstract: After-ripening (AR) is a time and environment regulated process occurring in the dry seed, which determines the germination potential of seeds. Both metabolism and perception of the phytohormone abscisic acid (ABA) are important in the initiation and maintenance of dormancy. However, molecular mechanisms that regulate the capacity for dormancy or germination through AR are unknown. To understand the relationship between ABA and AR, we analysed genome expression in Arabidopsis thaliana mutants defective in seed ABA synthesis (aba1-1) or perception (abi1-1). Even though imbibed mutant seeds showed no dormancy, they exhibited changes in global gene expression resulting from dry AR that were comparable with changes occurring in wild-type (WT) seeds. Core gene sets were identified that were positively or negatively regulated by dry seed storage. Each set included a gene encoding repression or activation of ABA function (LPP2 and ABA1, respectively), thereby suggesting a mechanism through which dry AR may modulate subsequent germination potential in WT seeds. Application of exogenous ABA to after-ripened WT seeds did not reimpose characteristics of freshly harvested seeds on imbibed seed gene expression patterns. It was shown that secondary dormancy states reinstate AR status-specific gene expression patterns. A model is presented that separates the action of ABA in seed dormancy from AR and dry storage regulated gene expression. These results have major implications for the study of genetic mechanisms altered in seeds as a result of crop domestication into agriculture, and for seed behaviour during dormancy cycling in natural ecosystems.

Effects of arsenic on seed germination and physiological activities of wheat seedlings.

Abstract: The effects of arsenic (As) were investigated on seed germination, root and shoot length and their biomass and some other factors to elucidate the toxicity of As. The results showed low concentrations of As (0–1 mg/kg) stimulated seed germination and the growth of root and shoot, however, these factors all decreased gradually at high concentrations of As (5–20 mg/kg). The contents of O 2 .− , MDA, soluble protein and peroxidase (POD) activity all increased with increasing As concentrations. Soluble sugar content, ascorbate peroxidase (APX), and superoxide dismutase (SOD) activities decreased at low concentrations of As, and increased at high concentrations of As. While acetylsalicylic acid (ASA) and chlorophyll contents, catalase (CAT) activity displayed increasing trend when the concentrations of As was lower than 1 mg/kg, and then decreasing trend. By polyacrylamide gel electrophoresis (PAGE), As induced the expression of POD isozymes of wheat seedlings. As induced the expression of CAT isozymes but inhibited the expression of SOD isozymes of wheat seedlings at concentrations lower than 1 mg/kg. However, As inhibited the expression of CAT isozymes but induced the expression of SOD isozymes at concentrations higher than 5 mg/kg. The results indicated As could exert harmfulness in the early development stage of wheat at inappropriate concentrations.

Functional characterization of a glycine-rich RNA-binding protein 2 in Arabidopsis thaliana under abiotic stress conditions

Abstract: Summary Although glycine-rich RNA-binding protein 2 (GRP2) has been implicated in plant responses to environmental stresses, the function and importance of GRP2 in stress responses are largely unknown. Here, we examined the functional roles of GRP2 in Arabidopsis thaliana under high-salinity, cold or osmotic stress. GRP2 affects seed germination of Arabidopsis plants under salt stress, but does not influence seed germination and seedling growth of Arabidopsis plants under osmotic stress. GRP2 accelerates seed germination and seedling growth in Arabidopsis plants under cold stress, and contributes to enhancement of cold and freezing tolerance in Arabidopsis plants. No differences in germination between the wild-type and transgenic plants were observed following addition of abscisic acid (ABA) or glucose, implying that GRP2 affects germination through an ABA-independent pathway. GRP2 complements the cold sensitivity of an Escherichia coli BX04 mutant and exhibits transcription anti-termination activity, suggesting that it has an RNA chaperone activity during the cold adaptation process. Mitochondrial respiration and catalase and peroxidase activities were affected by expression of mitochondrial-localized GRP2 in Arabidopsis plants under cold stress. Proteome analysis revealed that expression of several mitochondrial-encoded genes was modulated by GRP2 under cold stress. These results provide new evidence indicating that GRP2 plays important roles in seed germination, seedling growth and freezing tolerance of Arabidopsis under stress conditions, and that GRP2 exerts its function by modulating the expression and activity of various classes of genes.

Influence of germination on the nutritional quality of Phaseolus vulgaris and Cajanus cajan

Abstract: Germination is a method that can modify the presence of nutrients and antinutrients in legume seeds. In this study, the changes in trypsin inhibitors activity (TIA), phytic acid, tannins, ascorbic acid, thiamine, protein digestibility and minerals in germinated black beans (Phaseolus vulgaris L.), white beans (Phaseolus vulgaris L.) and pigeon beans (Cajanus cajan L. Millsp) were evaluated. The ungerminated grains were analysed as control. A significant decrease in the content of TIA of pigeon beans (19.2%), white beans (52.5%) and black beans (25%) was observed. The reduction of phytic acid was more than 40% for the three grains germinated; for the tannins, the reduction was of 14.3% for pigeon beans, 19% for black beans and 36.2% for white beans. Germination increased the protein digestibility in a 2–4% range, and also increased the ascorbic acid by 300% for white beans, by 33% for black beans and by 208% for pigeon beans. The thiamine content increased more than 26.7%. Germination affected the mineral content erratically, depending more on the grain and the type of mineral. These variations in the content of nutrients and antinutrients of the germinated grains are attributed to the joint effect of the germination and previous soaking the grains were subjected to.

Two B3 domain transcriptional repressors prevent sugar-inducible expression of seed maturation genes in Arabidopsis seedlings.

Abstract: During development of plant seeds, embryos import nutrients and store massive amounts of reserves. Seed reserves are rapidly degraded and mobilized to support seedling development after germination. HIGH-LEVEL EXPRESSION OF SUGAR-INDUCIBLE GENE 2 (HSI2) of Arabidopsis thaliana is a B3 DNA-binding domain protein that represses the transcription of sugar-inducible reporter gene. Although disruption of HSI2 or HSI2-Like 1 (HSL1) did not affect growth, seeds with disruption of both HSI2 and HSL1 (KK mutant) developed abortive seedlings that stopped growing 7–9 days after imbibition. KK seedlings developed swollen hypocotyls that accumulated seed storage proteins and oil on medium containing sucrose or other metabolizable sugars, and calluses developed from KK seedlings also accumulated seed storage reserves. The expression of seed maturation genes, which include LEAFY COTYLEDON-type master regulators, in KK seedlings depended on the concentration of sucrose, suggesting that sugar controls the expression of seed maturation genes. Our results suggest that HSI2 and HSL1 repress the sugar-inducible expression of the seed maturation program in seedlings and play an essential role in regulating the transition from seed maturation to seedling growth.

Size-Related Differential Seed Predation in a Heavily Defaunated Neotropical Rain Forest

Abstract: Hunting and habitat loss represent an increasingly common anthropogenic impact. Available evidence suggests that defaunation is typically differential: medium/large mammals are most affected while small rodents are less affected, or even favored. In heavily impacted areas, such as Los Tuxtlas, the residual mammalian fauna is dominated by small rodents. We tested the expectation that if small rodents will preferentially attack small-seeded species, large-seeded species may escape predation in the absence of medium/large seed predators and that this may affect germination and, possibly, recruitment. Experiments with caged rodents (Heteromys desmarestianus) under laboratory conditions showed a preference for smaller seeds. A field experiment involving seeds of contrasting size showed that small, unprotected seeds experienced a predation risk 30-times larger than protected seeds, while the effect of protection was not significant for large seeds. These patterns of predation led to significant differences in germination: protected small-seeded species had a fourfold greater germination than unprotected small-seeded species, while germination was not significantly different between exposed and protected large seeds. The observed contrasts in germination suggest that under heavy defaunation, small-seeded species are likely to be penalized by the overabundance of small rodents, while large-seeded species escape predation. Our results are consistent with the observation of dense seedling carpets dominated by large-seeded species in the understory of Los Tuxtlas. We speculate that the patterns we uncovered may explain, at least partly, the impoverished diversity of the understory vegetation that characterizes heavily defaunated understories and that this has the potential to affect forest regeneration.

Seed priming with brassinolide improves lucerne (Medicago sativa L.) seed germination and seedling growth in relation to physiological changes under salinity stress

Abstract: Salt stress is an important constraint to lucerne (Medicago sativa L.) production in many parts of the world. Seeds of 3 lucerne varieties, cvv. Victoria, Golden Empress, and Victor, were used to investigate the effects of seed priming with 5 µm/L brassinolide on germination and seedling growth under a high level of salt stress (13.6 dS/m NaCl solution). The results showed that germination percentage, germination index, and vigour index of lucerne seeds primed with brassinolide were significantly higher than those of the non-primed seeds under salinity stress in each variety. Seed priming with brassinolide significantly increased the shoot fresh weight, shoot dry weight, and root dry weight in 2 varieties, and significantly increased the root length and root vigour in each variety. It also significantly increased the activities of antioxidant enzymes, peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT), in Victoria and Victor seedlings. During seedling growth, the primed seeds significantly reduced the malondialdehyde (MDA) accumulation. This suggests that priming lucerne seed with brassinolide at a suitable concentration can improve germination and seedling growth under high-saline soils.

Orchid-associated bacteria produce indole-3-acetic acid, promote seed germination, and increase their microbial yield in response to exogenous auxin.

Abstract: Germination of orchid seeds is a complex process. In this paper we focus on interactions between the host-plant and its bacterial partners via indole-3-acetic acid (IAA). Originally isolated from the roots of the epiphytic orchid Dendrobium moschatum, the strains of Rhizobium, Microbacterium, Sphingomonas, and Mycobacterium genera were among the most active IAA producers. Addition of exogenous tryptophan significantly enhanced auxin formation both in mineral and complex media. The presence of IAA and indole-3-acetaldehyde was confirmed by HPLC. Indole-3-pyruvic and indole-3-lactic acids were also detected in supernatants of culture filtrates of Sphingomonas sp., Rhizobium sp., and Microbacterium sp., while indole-3-acetamide was identified only in Mycobacterium sp. Some concentration- and strain-dependent effects of exogenous IAA on bacterial development were also established. Treatment of the cultures with 10 and 100 μg/ml of auxin resulted in an increase in microbial yield. None of the investigated strains was able to utilize IAA as a source of carbon and energy. Furthermore, inoculation of D. moschatum seeds with Sphingomonas sp. and Mycobacterium sp. resulted in considerable enhancement of orchid seeds germination. This growth-promoting activity was observed in the absence of any plant growth stimulators or mycorrhizal fungi, usually required for orchid germination.

Performance of wheat genotypes under osmotic stress at germination and early seedling growth stage

Abstract: Sixteen wheat genotypes including local varieties were tested in completely randomized design with three repeats. Data were recorded at four different moisture levels by using polyethylene glycol (PEG) 6000 on germination percentage, germination rate index, shoot length, root length, fresh weight of shoot, dry weight of shoot, fresh weight of root, dry weight of root, shoot/root ratio and analysed for significance. The genotypes differ significantly in response to the moisture stress. There were highly significant differences for all traits. PK-18199 gave the maximum germination percentage, germination rate index, shoot length root length, coleoptile length, fresh shoot weight, dry shoot weight, fresh root weight, dry root weight and root/shoot ratio under all four moisture stresses. PK-18175 showed maximum resistance against moisture stress while WAFAQ 2001 showed minimum resistance. AS-2002 and KC033 also gave the better performance under all four moisture levels for most of the traits at seedling stage. 99FJ03 gave maximum root/shoot length ratio while PK 18199 gave minimum value of root/shoot length ratio showing resistance against water stress.

Reactive oxygen species, ABA and nitric oxide interactions on the germination of warm-season C4-grasses

Abstract: Hydrogen peroxide (H2O2) as a source of reactive oxygen species (ROS) signiWcantly stimulated ger- mination of switchgrass (Panicum virgatum L.) seeds with an optimal concentration of 20 mM at both 25 and 35°C. For non-dormant switchgrass seeds exhibiting diVerent levels of germination, treatment with H 2 O 2 resulted in rapid germination (<3 days) of all germinable seeds as compared to seeds placed on water. Exposure to 20 mM H 2 O 2 elicited simultaneous growth of the root and shoot system, resulting in more uniform seedling development. Seeds of big bluestem (Andropogon gerardii Vitman) and indiangrass (Sorghastrum nutans (L.) Nash) also responded positively to H2O2 treatment, indicating the universality of the eVect of H 2 O 2 on seed germination in warm-season prairie grasses. For switchgrass seeds, abscisic acid (ABA) and the NADPH-oxidase inhibitor, diphenyleneiodonium (DPI) at 20 M retarded germination (radicle emergence), stunted root growth and partially inhibited NADPH-oxidase activ- ity in seeds. H2O2 reversed the inhibitory eVects of DPI and ABA on germination and coleoptile elongation, but did not overcome DPI inhibition of root elongation. Treatment with H 2 O 2 appeared to enhance endogenous production of nitric oxide, and a scavenger of nitric oxide abolished the perox- ide-responsive stimulation of switchgrass seed germination. The activities and levels of several proteins changed earlier in seeds imbibed on H 2 O 2 as compared to seeds maintained on water or on ABA. These data demonstrate that seed ger- mination of warm-season grasses is signiWcantly responsive to oxidative conditions and highlights the complex inter- play between seed redox status, ABA, ROS and NO in this system.

Thioredoxin-Linked Proteins Are Reduced during Germination of Medicago truncatula Seeds

Abstract: Germination of cereals is accompanied by extensive change in the redox state of seed proteins. Proteins present in oxidized form in dry seeds are converted to the reduced state following imbibition. Thioredoxin (Trx) appears to play a role in this transition in cereals. It is not known, however, whether Trx-linked redox changes are restricted to cereals or whether they take place more broadly in germinating seeds. To gain information on this point, we have investigated a model legume, Medicago truncatula. Two complementary gel-based proteomic approaches were followed to identify Trx targets in seeds: Proteins were (1) labeled with a thiol-specific probe, monobromobimane (mBBr), following in vitro reduction by an NADP/Trx system, or (2) isolated on a mutant Trx affinity column. Altogether, 111 Trx-linked proteins were identified with few differences between axes and cotyledons. Fifty nine were new, 34 found previously in cereal or peanut seeds, and 18 in other plants or photosynthetic organisms. In parallel, the redox state of proteins assessed in germinating seeds using mBBr revealed that a substantial number of proteins that are oxidized or partly reduced in dry seeds became more reduced upon germination. The patterns were similar for proteins reduced in vivo during germination or in vitro by Trx. In contrast, glutathione and glutaredoxin were less effective as reductants in vitro. Overall, more than half of the potential targets identified with the mBBr labeling procedure were reduced during germination. The results provide evidence that Trx functions in the germination of seeds of dicotyledons as well as monocotyledons.

Characterization of Pythium spp. Associated with Corn and Soybean Seed and Seedling Disease in Ohio.

Abstract: Cool, moist conditions in combination with minimum tillage, earlier planting, and recent shifts in commercial fungicide seed-treatment active ingredients have led to an increase in corn (Zea mays) and soybean (Glycine max) seedling establishment problems. This situation resulted in an investigation of Pythium spp. associated with seed and seedling diseases. Samples of diseased corn and soybean seedlings were collected from 42 production fields in Ohio. All isolates of Pythium recovered were identified to species using morphological and molecular techniques and evaluated in an in vitro pathogenicity assay on both corn and soybean seed, and a subset of the isolates was tested for sensitivity to fungicides currently used as seed treatments. Eleven species and two distinct morphological groups of Pythium were identified, of which six species were moderately to highly pathogenic on corn seed and nine species were highly pathogenic on soybean seed. There was significant variation (P < 0.05) in sensitivity to mefenoxam, azoxystrobin, trifloxystrobin, and captan both across and within species. Multiple species of Pythium had the capacity to reduce germination of both corn and soybean seed. Results indicated that mefenoxam, azoxystrobin, trifloxystrobin, or captan, when used individually, may not inhibit all pathogenic species of Pythium found in Ohio soils.

Seed Treatment with Auxins Modulates Growth and Ion Partitioning in Salt-stressed Wheat Plants

Abstract: Experiments were performed to determine whether seed priming with different concentrations (100, 150, and 200 mg/L) of auxins (indoleacetic acid (IAA), indolebutyric acid (IBA), or their precursor tryptophane (Trp)) could alter salinity induced perturbances in salicylic acid and ion concentrations and, hence, growth in wheat (Triticum aestivum L.) cultivars, namely M.H.-97 (salt intolerant) and Inqlab-91 (salt tolerant). Primed and non-primed seeds were sown in Petri dishes in a growth room, as well as in a field treated with 15 dS/m NaCl salinity. All priming agents, except IBA, increased the final germination percentage in both cultivars. The seedlings of either cultivar raised from Trp-treated seeds had greater dry biomass when under salt stress. In field experiments, Trp priming was much more effective in mediating the increase in grain yield, irrespective of the cultivar, under salt stress. The alleviatory effect of Trp was found to be associated with reduced uptake of Na + in the roots and subsequent translocation to the shoots, as well as increased partitioning of Ca 2+ in the roots of salt-stressed wheat plants. Plants of both cultivars raised from Trp- and IAA-treated seeds accumulated free salicylic acid in their leaves when under salt stress. Overall, the Trp priming-induced improvement in germination and the subsequent growth of wheat plants could be related to ion homeostasis when under salt stress. The possible involvement of salicylic acid in the Trp priming-induced better growth under conditions of salt stress is discussed.

Impact of red : far red ratios on germination of temperate forest herbs in relation to shade tolerance, seed mass and persistence in the soil

Abstract: Summary 1The effect of the ratio of red : far red light (R : FR) on seed germination of herbaceous species from northern temperate deciduous forest has received little attention Here for 27 such species, we investigated the relationship between seed mass and the suitability of micro-sites for germination Germination in light and in darkness was compared after cold stratification, and for the light-requiring species, the germination response to R : FR was determined In addition, seed bank persistence was monitored over 4 years 2With increasing seed mass, germination became less dependent on light and seed bank persistence decreased Furthermore, for the light-dependent species, there was a significant negative relationship between the R : FR that resulted in 50% germination and seed mass 3These data suggest that small-seeded species only germinate in micro-sites with a high R : FR, which signals the absence of over-topping vegetation or leaf litter Such micro-sites are comparatively rare, which may necessitate persistence in the soil seed bank 4For small-seeded species, we propose that a key benefit from restricting germination to high R : FR is a reduction in the time-frame over which the initially small seedlings remain small and hence highly vulnerable to mortality: high R : FR is likely to indicate high-light levels which would facilitate rapid seedling growth

The role of water channel proteins and nitric oxide signaling in rice seed germination

Abstract: Previous studies have demonstrated the possible role of several aquaporins in seed germination. But systematic investigation of the role of aquaporin family members in this process is lacking. Here, the developmental regulation of plasma membrane intrinsic protein (PIP) expression throughout germination and post-germination processes in rice embryos was analyzed. The expression patterns of the PIPs suggest these aquaporins play different roles in seed germination and seedling growth. Partial silencing of the water channel genes, OsPIP1;1 and OsPIP1;3, reduced seed germination while over-expression of OsPIP1;3 promoted seed germination under water-stress conditions. Moreover, spatial expression analysis indicates that OsPIP1;3 is expressed predominantly in embryo during seed germination. Our data also revealed that the nitric oxide (NO) donors, sodium nitroprusside (SNP) and S-nitrosoglutathione (GSNO), promoted seed germination; furthermore, the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, inhibited germination and reduced the stimulative effects of SNP and GSNO on rice germination. Exogenous NO stimulated the transcription of OsPIP1;1, OsPIP1;2, OsPIP1;3 and OsPIP2;8 in germinating seeds. These results suggest that water channels play an important role in seed germination, acting, at least partly, in response to the NO signaling pathway.