Ethylene signaling involves in seeds germination upon submergence and antioxidant response elicited confers submergence tolerance to rice seedlings.
ABSTRACT: BACKGROUND:Flooding has negative impact on agriculture. The plant hormone ethylene is involved in plant growth and stress responses, which are important role in tolerance and adaptation regulatory mechanisms during submergence stress. Ethylene signaling crosstalk with gibberellin signaling enhances tolerance in lowland rice (Flood Resistant 13A) through a quiescence strategy or in deepwater rice through an escape strategy when rice is submerged. Information regarding ethylene-mediated priming in submergence stress tolerance in rice is scant. Here, we used 1-aminocyclopropane-1-carboxylic acid, an ethylene precursor, to evaluate the response in submerged rice seedlings. RESULTS:The germination rate and mean germination times of rice seeds was higher in seedlings under submergence only when ethylene signaling was inhibited by supplemented with silver nitrate (AgNO3). Reduced leaf chlorophyll contents and induced senescence-associated genes in rice seedlings under submergence were relieved by pretreatment with an ethylene precursor. The ethylene-mediated priming by pretreatment with an ethylene precursor enhanced the survival rate and hydrogen peroxide (H2O2) and superoxide (O2-) anion accumulation and affected antioxidant response in rice seedlings. CONCLUSIONS:Pretreatment with an ethylene precursor leads to reactive oxygen species generation, which in turn triggered the antioxidant response system, thus improving the tolerance of rice seedlings to complete submergence stress. Thus, H2O2 signaling may contribute to ethylene-mediated priming to submergence stress tolerance in rice seedlings.
Project description:Submergence stress is a limiting factor for direct-seeded rice systems in rainfed lowlands and flood-prone areas of South and Southeast Asia. The present study demonstrated that submergence stress severely hampered the germination and seedling growth of rice, however, seed priming alleviated the detrimental effects of submergence stress. To elucidate the molecular basis of seed priming-induced submergence tolerance, transcriptome analyses were performed using 4-day-old primed (selenium-Se and salicylic acid-SA priming) and non-primed rice seedlings under submergence stress. Genomewide transcriptomic profiling identified 2371 and 2405 transcripts with Se- and SA-priming, respectively, that were differentially expressed in rice compared with non-priming treatment under submergence. Pathway and gene ontology term enrichment analyses revealed that genes involved in regulation of secondary metabolism, development, cell, transport, protein, and metal handling were over-represented after Se- or SA-priming. These coordinated factors might have enhanced the submergence tolerance and maintained the better germination and vigorous seedling growth of primed rice seedlings. It was also found that many genes involved in cellular and metabolic processes such as carbohydrate metabolism, cellular, and metabolic biosynthesis, nitrogen compound metabolic process, transcription, and response to oxidative stress were induced and overlapped in seed priming treatments, a finding which reveals the common mechanism of seed priming-induced submergence tolerance. Taken together, these results may provide new avenues for understanding and advancing priming-induced responses to submergence tolerance in crop plants.
Project description:Rice (Oryza sativa L.) is the only cereal that can be cultivated in the frequently flooded river deltas of South-East and South Asia. The survival strategies used by rice have been studied quite extensively and the role of several phytohormones in the elongation response has been established. Deep-water rice cultivars can diminish flooding stress by rapid elongation of their submerged tissues to keep up with the rising waters. Other rice cultivars may react by mechanisms of submergence tolerance. Aerenchyma and aerenchymatous adventitious roots are formed that facilitate oxygen diffusion to prevent anaerobic conditions in the submerged tissues. This paper discusses the molecular aspects of the mechanism that leads to shoot elongation (leaves of seedlings and internodes), the regulation of which involves metabolism of, and interactions between, ethylene, gibberellins and abscisic acid. Finally, the importance of new techniques in future research is assessed. Current molecular technology can reveal subtle differences in gene activity between tolerant and non-tolerant cultivars, and identify genes that are involved in the regulation of submergence avoidance and tolerance.
Project description:Submergence tolerance of 13 doubled haploid lines of rice and their parents (submergence tolerant FR13A and submergence intolerant CT6241) was assessed using 2-week-old seedlings. Plants were scored for leaf senescence and percentage of seedlings that survived up to 15 d submergence, followed by a 12 d recovery period. Seven lines proved to be submergence tolerant, and six relatively intolerant. In all lines, activity of pyruvate decarboxylase (PDC), extracted from the apical 3-5 cm of root axes, decreased by 46-96 % and 38-76 %, respectively, during 5 or 10 d submergence under natural day/night conditions, compared with pre-submergence values (100 %). However, when the enzyme was extracted at night, submergence increased PDC activity of all rice lines (approx. 112 % on average), compared with pre-submergence values (100 %). The stimulating effect of the dark period on PDC activity was reproduced and amplified by submerging rice seedlings for up to 5 d in continuous darkness in water containing sub-ambient concentrations of oxygen (2.3 mg l(-1)). Such increased PDC activity was also observed in seedlings exposed to anoxia for 6 h (approx. 6-175 % higher than pre-submergence values). Irrespective of tolerance class, submergence decreased soluble protein concentrations under all conditions and sampling times. No positive correlation was found between PDC activity and tolerance of the various rice lines to submergence. However, PDC activity was slightly higher in submergence intolerant lines, compared with tolerant lines, under both dark submergence and anoxia. Such differences in PDC activity between the two groups of rice lines were not observed when they were submerged under the natural diurnal cycle. Increased PDC activity in roots at night demonstrated a probable incidence of tissue hypoxia or anoxia during submergence during each dark period.
Project description:Acireductone dioxygenase (ARD) is a metal-binding metalloenzyme and involved in the methionine salvage pathway. In rice, OsARD1 binds Fe2+ and catalyzes the formation of 2-keto-4-methylthiobutyrate (KMTB) to produce methionine, which is an initial substrate in ethylene synthesis pathway. Here, we report that overexpression of OsARD1 elevates the endogenous ethylene release rate, enhances the tolerance to submergence stress, and reduces the sensitivity to drought, salt, and osmotic stresses in rice. OsARD1 is strongly induced by submergence, drought, salinity, PEG6000, and mechanical damage stresses and exhibits high expression level in senescent leaves. Transgenic plants overexpressing OsARD1 (OsARD1-OE) display fast elongation growth to escape submergence stress. The ethylene content is significantly maximized in OsARD1-OE plants compared with the wide type. OsARD1-OE plants display increased shoot elongation and inhibition of root elongation under the submergence stress and grow in dark due to increase of ethylene. The elongation of coleoptile under anaerobic germination is also significantly promoted in OsARD1-OE lines due to the increase of ethylene content. The sensitivity to drought and salt stresses is reduced in OsARD1-OE transgenic lines. Water holding capacity is enhanced, and the stomata and trichomes on leaves increase in OsARD1-OE lines. Drought and salt tolerance and ethylene synthesis-related genes are upregulated in OsARD1-OE plants. Subcellular localization shows that OsARD1 displays strong localization signal in cell nucleus, suggesting OsARD1 may interact with the transcription factors. Taken together, the results provide the understanding of the function of OsARD1 in ethylene synthesis and abiotic stress response in rice.
Project description:Ethylene is gaseous plant hormone that controls a variety of physiologic activities. OsERS1 and OsETR2 are major ethylene receptors in rice that have been reported to have different regulatory functions. The GFP fused N-terminus of OsERS1 and OsETR2 showed differentially localization patterns when transiently expressed in onion epidermal cells. Base on these results, we suggested that OsERS1 could be localized to plasma membranes, whereas OsETR2 could be localized to the endoplasmic reticulum. Furthermore, instead of the constitutive expression profile of OsERS1, OsETR2 is differentially expressed in seedlings of light/dark-grown conditions, submergence or exogenous ethylene treatments. Our results and others support the notion that OsERS1 and OsETR2 could have different roles during rice plant submergence.
Project description:Submergence tolerance is an important agronomic trait for rice grown in South-East Asia, where flash flooding occurs frequently and unpredictably during the monsoons. Although mapping locations of one major and several minor quantitative trait loci (QTL) were known previously, improving submergence tolerance in agronomically desirable types of rice has not been achieved. KDML105 is jasmine rice widely grown in rain-fed lowland regions of Thailand. This cultivar is very intolerant of submergence stress. To improve submergence tolerance in this cultivar, three submergence-tolerant cultivars, FR13A, IR67819F2-CA-61 and IR49830-7-1-2-2, were cross-pollinated with KDML105. Transferring the major QTL for submergence tolerance was facilitated by four back-crossings to the recipient KDML105. Molecular markers tightly linked to the gene(s) involved were developed to facilitate molecular genotyping. We demonstrated that individuals of a BC4F3 line that retained a critical region on chromosome 9 transferred from tolerant lines were also tolerant of complete submergence while retaining all the agronomically desirable traits of KDML105. In addition, effects of secondary QTLch2 were detected statistically in back-cross progenies. Effects of secondary QTLch7 were not statistically significant. The close association between tightly linked markers of the tolerance locus on chromosome 9 and submergence tolerance in the field demonstrates the considerable promise of using these markers in lowland rice breeding programmes for selecting increased submergence tolerance.
Project description:Rice plants are much damaged by several days of total submergence. The effect can be a serious problem for rice farmers in the rainfed lowlands of Asia, and runs contrary to a widespread belief amongst plant biologists that rice is highly tolerant of submergence. This article assesses the characteristics of the underwater environment that may damage rice plants, examines various physiological mechanisms of injury, and reviews recent progress achieved using linkage mapping to locate quantitative traits loci (QTL) for tolerance inherited from a submergence-tolerant cultivar FR13A. Progress towards identifying the gene(s) involved through physical mapping of a dominant tolerance locus on chromosome 9 is also summarized. Available physiological evidence points away from responses to oxygen shortage as being inextricably involved in submergence injury. An imbalance between production and consumption of assimilates is seen as being especially harmful, and is exacerbated by strongly accelerated leaf extension and leaf senescence that are ethylene-mediated and largely absent from FR13A and related cultivars. DNA markers for a major QTL for tolerance are shown to be potentially useful in breeding programmes designed to improve submergence tolerance.
Project description:BACKGROUND:Flooding during seasonal monsoons affects millions of hectares of rice-cultivated areas across Asia. Submerged rice plants die within a week due to lack of oxygen, light and excessive elongation growth to escape the water. Submergence tolerance was first reported in an aus-type rice landrace, FR13A, and the ethylene-responsive transcription factor (TF) gene SUB1A-1 was identified as the major tolerance gene. Intolerant rice varieties generally lack the SUB1A gene but some intermediate tolerant varieties, such as IR64, carry the allelic variant SUB1A-2. Differential effects of the two alleles have so far not been addressed. As a first step, we have therefore quantified and compared the expression of nearly 2500 rice TF genes between IR64 and its derived tolerant near isogenic line IR64-Sub1, which carries the SUB1A-1 allele. Gene expression was studied in internodes, where the main difference in expression between the two alleles was previously shown. RESULTS:Nineteen and twenty-six TF genes were identified that responded to submergence in IR64 and IR64-Sub1, respectively. Only one gene was found to be submergence-responsive in both, suggesting different regulatory pathways under submergence in the two genotypes. These differentially expressed genes (DEGs) mainly included MYB, NAC, TIFY and Zn-finger TFs, and most genes were downregulated upon submergence. In IR64, but not in IR64-Sub1, SUB1B and SUB1C, which are also present in the Sub1 locus, were identified as submergence responsive. Four TFs were not submergence responsive but exhibited constitutive, genotype-specific differential expression. Most of the identified submergence responsive DEGs are associated with regulatory hormonal pathways, i.e. gibberellins (GA), abscisic acid (ABA), and jasmonic acid (JA), apart from ethylene. An in-silico promoter analysis of the two genotypes revealed the presence of allele-specific single nucleotide polymorphisms, giving rise to ABRE, DRE/CRT, CARE and Site II cis-elements, which can partly explain the observed differential TF gene expression. CONCLUSION:This study identified new gene targets with the potential to further enhance submergence tolerance in rice and provides insights into novel aspects of SUB1A-mediated tolerance.
Project description:Submergence-tolerant rice maintains viability during complete submergence by limiting underwater elongation until floodwaters recede. Acclimation responses to submergence are coordinated by the submergence-inducible Sub1A, which encodes an ethylene-responsive factor-type transcription factor (ERF). Sub1A is limited to tolerant genotypes and sufficient to confer submergence tolerance to intolerant accessions. Here we evaluated the role of Sub1A in the integration of ethylene, abscisic acid (ABA), and gibberellin (GA) signaling during submergence. The submergence-stimulated decrease in ABA content was Sub1A-independent, whereas GA-mediated underwater elongation was significantly restricted by Sub1A. Transgenics that ectopically express Sub1A displayed classical GA-insensitive phenotypes, leading to the hypothesis that Sub1A limits the response to GA. Notably Sub1A increased the accumulation of the GA signaling repressors Slender Rice-1 (SLR1) and SLR1 Like-1 (SLRL1) and concomitantly diminished GA-inducible gene expression under submerged conditions. In the Sub1A overexpression line, SLR1 protein levels declined under prolonged submergence but were accompanied by an increase in accumulation of SLRL1, which lacks the DELLA domain. In the presence of Sub1A, the increase in these GA signaling repressors and decrease in GA responsiveness were stimulated by ethylene, which promotes Sub1A expression. Conversely, ethylene promoted GA responsiveness and shoot elongation in submergence-intolerant lines. Together, these results demonstrate that Sub1A limits ethylene-promoted GA responsiveness during submergence by augmenting accumulation of the GA signaling repressors SLR1 and SLRL1.
Project description:Flash flooding of young rice plants is a common problem for rice farmers in south and south-east Asia. It severely reduces grain yield and increases the unpredictability of cropping. The inheritance and expression of traits associated with submergence stress tolerance at the seedling stage are physiologically and genetically complex. We exploited naturally occurring differences between certain rice lines in their tolerance to submergence and used quantitative trait loci (QTL) mapping to improve understanding of the genetic and physiological basis of submergence tolerance. Three rice populations, each derived from a single cross between two cultivars differing in their response to submergence, were used to identify QTL associated with plant survival and various linked traits. These included total shoot elongation under water, the extent of stimulation of shoot elongation caused by submergence, a visual submergence tolerance score, and leaf senescence under different field conditions, locations and years. Several major QTL determining plant survival, plant height, stimulation of shoot elongation, visual tolerance score and leaf senescence each mapped to the same locus on chromosome 9. These QTL were detected consistently in experiments across all years and in the genetic backgrounds of all three mapping populations. Secondary QTL influencing tolerance were also identified and located on chromosomes 1, 2, 5, 7, 10 and 11. These QTL were specific to particular traits, environments, or genetic backgrounds. All identified QTL contributed to increased submergence tolerance through their effects on decreased underwater shoot elongation or increased maintenance of chlorophyll levels, or on both. These findings establish the foundations of a marker-assisted scheme for introducing submergence tolerance into agriculturally desirable cultivars of rice.