Chlorophyll fluorescence parameters,CO2 photosynthetic rate and regeneration capacity as a result of complete submergence and subsequent re-emergence in rice (Oryza sativa L.)

Regeneration capacity of submerged rice (Oryza sativa) seedlings in terms of CO₂ photosynthetic rate, chlorophyll a fluorescence and chlorophyll and carbohydrate content were investigated in three Indica rice cultivars namely FR 13A, Kalaputia and IR 42 that differed in submergence tolerance. Twenty-one day old plants were completely submerged under water for 8 days. Subsequently, plants were kept under normal conditions with 5–10 cm of stagnant water above soil surface for a further period of 15 days. After complete submergence, all genotypes showed inhibition of CO₂ photosynthetic rate. Submergence treatment resulted in a significant reduction of Rubisco activity. Maximal photochemical efficiency (F_v/F_m) of PS II and area above the fluorescence curve between F_o and F_m decreased more under submergence especially in susceptible cultivar IR 42. When re-aerated, the plants recovered to various degrees. The carbohydrate content of plants was found to be significantly and positively associated with submergence tolerance and regeneration growth. The tolerant cultivar (FR 13A) could survive submergence apparently because it possessed 1.9–2.0 and 3.2–3.7-fold more non-structural carbohydrate content before and after submergence compared to the susceptible cultivar (IR 42) and it had a better capability to restore its photosynthetic capacity during post-submergence periods.     

淹水对三峡库区消落带香附子生理特性的影响

为了阐明香附子对三峡库区消落带水淹的耐淹机理,该研究模拟三峡库区消落带夏季汛期水淹环境,设置常规水分管理(CK)、根部水淹(T1)、半淹(T2)以及全淹(T3)4个不同处理组,研究香附子对不同淹水环境的生理响应机制。结果显示:(1)经过45d的水淹处理,所有水淹处理香附子叶片叶绿素a、叶绿素b、类胡萝卜素以及总叶绿素的含量均显著下降,但叶绿素a/b值仍维持在CK水平;(2)淹水并没有增加T1和T3处理植株可溶性蛋白、脯氨酸和可溶性糖含量,但显著增加了T2植株叶片脯氨酸和可溶性糖的含量;(3)随着水淹时间的延长,各水淹处理植株叶片丙二醛含量逐渐升高,且自水淹15d后均与CK植株差异显著;(4)所有水淹处理香附子植株SOD、CAT和APX等抗氧化酶活性均有所增加,但随着水淹时间的继续延长,T3植株叶片SOD和CAT活性逐渐下降。(5)经过45d的水淹处理,各水淹处理植株淀粉含量在叶和根中均显著增加。研究表明,虽然淹水对香附子产生了一定的伤害,但香附子能够通过调节体内各种保护酶活性和渗透调节物质含量来增加对水淹的抵御能力,而且根部高浓度淀粉含量也为香附子耐受水淹胁迫提供了稳定的能量供应。     

Submergence effects on rice genotypes during seedling stage: Probing of submergence driven changes of photosystem 2 by chlorophyll a fluorescence induction O-J-I-P transients

The effects of submergence on chlorophyll (Chl) a fluorescence were compared in seven Oryza sativa (L.) cultivars, namely FR 13A, Khoda, Khadara, Kalaputia (tolerant), Sabita, and Hatipanjari (avoiding type), and IR 42 (susceptible). Seedlings were submerged for 4 d under complete darkness. Oxygen concentration of flood water decreased with the period of submergence with concomitant increase in concentration of carbon dioxide. Submergence caused diminution in the amount of total Chl. Genotypic differences were observed for Chl content and survival percentage. Quantification of the Chl a fluorescence transients (JIP-test) revealed large cultivar differences in the response of photosystem 2 (PS2) to submergence. The kinetics of Chl a fluorescence rise showed complex changes in the magnitudes and rise of O-J, J-I, and I-P phases caused by submergence. The selective suppression of the J-I phase of fluorescence especially after 2 d of submergence provided evidence for weakened electron donation from the oxygen evolving complex whereas under severe submergence stress (4 d) both O-J and J-I steps were suppressed greatly with highly suppressed P-step, which resulted in lowering of variable fluorescence. Grouping probability or energetic connectivity between PS2 obtained through JIP-test from the data after 2 d of submergence showed a direct relation with survival percentage, i.e. fluorescence measurements contained the information of the survival chance of a plant under submerged conditions. The information could be used in identifying the submergence tolerant cultivars when the damage is not very severe.     

Physiology,gene expression,and metabolome of two wheat cultivars with contrasting submergence tolerance

Responses of wheat (Triticum aestivum) to complete submergence are not well understood as research has focused on waterlogging (soil flooding). The aim of this study was to characterize the responses of 2 wheat cultivars differing vastly in submergence tolerance to test if submergence tolerance was linked to shoot carbohydrate consumption as seen in rice. Eighteen‐day‐old wheat cultivars Frument (intolerant) and Jackson (tolerant) grown in soil were completely submerged for up to 19 days while assessing responses in physiology, gene expression, and shoot metabolome. Results revealed 50% mortality after 9.3 and 15.9 days of submergence in intolerant Frument and tolerant Jackson, respectively, and significantly higher growth in Jackson during recovery. Frument displayed faster leaf degradation as evident from leaf tissue porosity, chlorophyll_a, and metabolomic fingerprinting. Surprisingly, shoot soluble carbohydrates, starch, and individual sugars declined to similarly low levels in both cultivars by day 5, showing that cultivar Jackson tolerated longer periods of low shoot carbohydrate levels than Frument. Moreover, intolerant Frument showed higher levels of phytol and the lipid peroxidation marker malondialdehyde relative to tolerant Jackson. Consequently, we propose to further investigate the role of ethylene sensitivity and deprivation of reactive O₂ species in submerged wheat.     

LcFIN2, a novel chloroplast protein gene from sheepgrass,enhances tolerance to low temperature in Arabidopsis and rice

Adverse environmental stresses affect plant growth and crop yields. Sheepgrass (Leymus chinensis (Trin.) Tzvel), an important forage grass that is widely distributed in the east of Eurasia steppe, has high tolerance to extreme low temperature. Many genes that respond to cold stress were identified in sheepgrass by RNA‐sequencing, but more detailed studies are needed to dissect the function of those genes. Here, we found that LcFIN2, a sheepgrass freezing‐induced protein 2, encoded a chloroplast‐targeted protein. Expression of LcFIN2 was upregulated by freezing, chilling, NaCl and abscisic acid (ABA) treatments. Overexpression of LcFIN2 enhanced the survival rate of transgenic Arabidopsis after freezing stress. Importantly, heterologous expression of LcFIN2 in rice exhibited not only higher survival rate but also accumulated various soluble substances and reduced membrane damage in rice under chilling stress. Furthermore, the chlorophyll content, the quantum photochemistry efficiency of photosystem II (ΦPSII), the non‐photochemical quenching (NPQ), the net photosynthesis rate (Pn) and the expression of some chloroplast ribosomal‐related and photosynthesis‐related genes were higher in the transgenic rice under chilling stress. These findings suggested that the LcFIN2 gene could potentially be used to improve low‐temperature tolerance in crops.     

Physiological responses of different olive genotypes to drought conditions

Gas exchange rates, chlorophyll fluorescence, pressure–volume relationships, photosynthetic pigments, total soluble sugars, starch, soluble proteins and proline concentrations were investigated in five Olea europaea L. cultivars with different geographical origins (Arbequina, Blanqueta, Cobran?osa, Manzanilla and Negrinha) grown under Mediterranean field conditions. We found considerable genotypic differences among the cultivars. Comparing the diurnal gas exchange rates, we observed that Cobran?osa, Manzanilla and Negrinha had high photosynthetic rate than Arbequina and Blanqueta. The first group reveals to be better acclimated to drought conditions, and appears to employ a prodigal water-use strategy, whereas Blanqueta and Arbequina, with high water-use efficiency, appear to employ a conservative water-use strategy. The degree of midday depression in photosynthesis was genotype dependent, with a maximum in Arbequina and a minimum in Negrinha. The reductions in the photosynthetic rate were dependent from both stomatal and non-stomatal limitations. Elastic adjustment plays an important role as drought tolerance mechanism. The group of cultivars that employ a prodigal water-use strategy revealed high tissue elasticity, whereas Arbequina and Blanqueta revealed high tissue rigidity. We also identified the existence of drought tolerance mechanisms associated with soluble proteins accumulation in the foliage. The high levels of soluble proteins in Arbequina may represent an increased activity of oxidative stress defence enzymes and may also represent a reserve for post stress recovery. In all cultivars, especially in Manzanilla, free proline was accumulated in the foliage. The discussed aspects of drought stress metabolism may have an adaptative meaning, supporting the hypothesis that olive cultivars native to dry regions, such as Cobran?osa, Manzanilla and Negrinha, have more capability to acclimate to drought conditions than cultivars originated in regions with a more temperate climate, like Arbequina and Blanqueta.     

The key regulator of submergence tolerance,SUB1A,promotes photosynthetic and metabolic recovery from submergence damage in rice leaves

The submergence‐tolerance regulator, SUBMERGENCE1A (SUB1A), of rice (Oryza sativa L.) modulates gene regulation, metabolism and elongation growth during submergence. Its benefits continue during desubmergence through protection from reactive oxygen species and dehydration, but there is limited understanding of SUB1A's role in physiological recovery from the stress. Here, we investigated the contribution of SUB1A to desubmergence recovery using the two near‐isogenic lines, submergence‐sensitive M202 and tolerant M202(Sub1). No visible damage was detected in the two genotypes after 3 d of submergence, but the sublethal stress differentially altered photosynthetic parameters and accumulation of energy reserves. Submergence inhibited photosystem II photochemistry and stimulated breakdown of protein and accumulation of several amino acids in both genotypes at similar levels. Upon desubmergence, however, more rapid return to homeostasis of these factors was observed in M202(Sub1). Submergence considerably restrained non‐photochemical quenching (NPQ) in M202, whereas the value was unaltered in M202(Sub1) during the stress. Upon reaeration, submerged plants encounter sudden exposure to higher light. A greater capability for NPQ‐mediated photoprotection can benefit the rapid recovery of photosynthetic performance and energy reserve metabolism in M202(Sub1). Our findings illuminate the significant role of SUB1A in active physiological recovery upon desubmergence, a component of enhanced tolerance to submergence.     

外源调节剂对淹涝水稻幼苗株高及碳水化合物消耗的影响

为研究淹涝条件下水稻幼苗株高及碳水化合物消耗对不同外源生长调节剂的响应,本试验选用籼型常规稻IR64和导入耐淹涝基因Sub1的IR64-Sub1为试验材料,秧龄20 d时喷施1-氨基环丙烷-1-羧酸(ACC)、多效唑(PB)、赤霉素(GA)3种外源生长调节剂,以喷施清水为对照(CK).喷施处理2 d后进行0、4、8、12、16 d没顶淹涝胁迫,淹涝胁迫结束后常温恢复7 d,取样分析不同外源生长调节剂对水稻成活率、株高、叶绿素降解及恢复、地上部非结构性碳水化合物(NSC)消耗的影响.结果表明:淹涝导致水稻幼苗株高显著增长,叶片SPAD值快速下降,叶片可溶性糖迅速消耗,但耐淹涝品种IR64-Sub1淹水前茎鞘NSC含量明显高于IR64,淹涝中NSC消耗速率低于IR64,淹水结束后地上部淀粉含量高于IR64.外源PB处理显著抑制水稻幼苗株高增长、叶绿素降解及NSC消耗,提高存活率,且对IR64-Sub1效果更为显著.外源GA处理水稻幼苗叶绿素降解、株高增长和NSC消耗最快,植株恢复能力最低,耐淹涝能力最差,但与IR64相比,GA对IR64-Sub1淹涝耐性的抑制明显减弱.外源ACC促进伸长效果明显低于外源GA处理.淹水前喷施PB可有效抑制植株水下伸长,延缓叶绿素降解,减缓NSC消耗,保留更多NSC,为淹水胁迫解除后水稻快速恢复提供有利条件,这对于易涝地区减轻涝渍危害具有重要意义.     

Mechanism associated with nonstructural carbohydrate accumulation in submergence tolerant rice (Oryza sativa L.) cultivars

Nonstructural carbohydrate (NSC) accumulation in submergence tolerant rice cultivars (cv) was studied in six Indica rice [Oryza sativa (L.)] cv under control and simulated submerged conditions. Tolerant cultivars accumulated greater contents of NSC compared to the susceptible cultivars. Starch and total NSC content showed significant positive association with survival percentage. On the other hand, elongation due to submergence was significantly a negative association with survival. The CO₂ photosynthetic rate, chlorophyll content, maximum photochemical efficiency of PS II (Fv/Fm), and activities of Rubisco were not significantly different between tolerant and susceptible cv under control condition. The ADP glucose pyrophosphorylase (AGPPase) activity was significantly higher in the tolerant cv and was a positive association with starch/NSC, whereas Fructose 1,6-diphosphatase (FDPase) activity was significantly higher in susceptible cv compared to tolerant cv and was a negative association with starch/NSC. Greater activities of AGPPase along with lower activities of FDPase might facilitate greater accumulation of NSC in tolerant rice cultivars.     

Examination of two lowland rice cultivars reveals that gibberellin-dependent early response to submergence is not necessarily mediated by ethylene

Using two lowland rice (Oryza sativa L.) cultivars we found that in both cases submerged-induced elongation early after germination depends on gibberellins (GAs). Submergence increases the content of the active GA₁ by enhancing the expression of GA biosynthesis genes, thus facilitating the seedlings to escape from the water and preventing asphyxiation. However, the two cultivars differ in their response to ethylene. The cultivar Senia (short), by contrast to cultivar Bomba (tall), does not elongate after ethylene application, and submerged-induced elongation is not negated by an inhibitor of ethylene perception. Also, while ethylene emanation in Senia is not altered by submergence, Bomba seedlings emanate more ethylene upon desubmergence, associated with enhanced expression of the ethylene biosynthesis gene OsACS5. The cultivar Senia thus allows the possibility of clarifying the role of ethylene and other factors as triggers of GA biosynthesis enhancement in rice seedlings under submergence.Key words: gibberellins, ethylene, submergence, lowland rice, Oryza sativa     

Effect of submergence on translocation,starch content and amylolytic activity in deep-water rice

Submergence induces rapid internodal elongation in deep-water rice (Oryza sativa L. cv. Habiganj Aman II). We investigated the metabolic activities which help to support such fast growth. Three days of submergence in water under continuous light led to the mobilization of 65% of the starch from those regions of rice internodes which had been formed prior to submergence. Disappearance of starch was accompanied by a 70-fold enhancement of amylolytic activity. Similar increases in amylolytic activity were detected in response to ethylene and gibberellic acid. Submergence also caused a 26-fold increase in the translocation of newly synthesized photosynthetic assimilates from the leaves to the internodes and younger regions of the culms. These physiological processes are likely to provide the metabolic energy required for internodal elongation in response to submergence.Abbreviation GA₃ gibberellic acid     

Light acclimation, CO2 response and long-term capacity of underwater photosynthesis in three terrestrial plant species

To characterize underwater photosynthetic performance in some terrestrial plants, we determined (i) underwater light acclimation (ii) underwater photosynthetic response to dissolved CO₂, and (iii) underwater photosynthetic capacity during prolonged submergence in three species that differ in submergence tolerance: Phalaris arundinacea, Rumex crispus (both submergence-tolerant) and Arrhenatherum elatius (submergence-intolerant). None of the species had adjusted to low irradiance after 1 week of submergence. Under non-submerged (control) conditions, only R. crispus displayed shade acclimation. Submergence increased the apparent quantum yield in this species, presumably because of the enhanced CO₂ affinity of the elongated leaves. In control plants of the grass species P. arundinacea and A. elatius, CO₂ affinities were higher than for R. crispus. The underwater photosynthetic capacity of R. crispus increased during 1 month of submergence. In P. arundinacea photosynthesis remained constant during 1 month of submergence at normal irradiance; at low irradiance a reduction in photosynthetic capacity was observed after 2 weeks, although there was no tissue degeneration. In contrast, underwater photosynthesis of the submergence-intolerant species A. elatius collapsed rapidly under both irradiances, and this was accompanied by leaf decay. To describe photosynthesis versus irradiance curves, four models were evaluated. The hyperbolic tangent produced the best goodness-of-fit, whereas the rectangular hyperbola (Michaelis-Menten model) gave relatively poor results.     

Suppression of gravitropic response of primary roots by submergence

Primary roots of six plant species were placed horizontally either in humid air or under water, and their growth and gravitropic responses were examined. In air, all the roots showed a normal gravitropic curvature. Under water without aeration, roots of rice (Oryza sativa L.), oat (Avena sativa L.), azuki bean (Vigna angularis Ohwi et Ohashi), and cress (Lepidium sativum L.) curved downward at almost same rate as in air, whereas the curvature of roots of maize (Zea mays L.) and pea (Pisum sativum L.) was strongly suppressed. Submergence did not cause a decrease in growth rate of these roots. When roots of maize and pea were placed horizontally under water without aeration and then rotated in three dimensions on a clinostat in air, they showed a significant curvature, suggesting that the step suppressed by submergence is not graviperception but the subsequent signal transmission or differential growth process. Constant bubbling of air through the water partly restored the gravitropic curvature of maize roots and completely restored that of pea roots. The curvature of pea roots was also partly restored by the addition of an inhibitor of ethylene biosynthesis, aminooxyacetic acid. In air, ethylene suppressed the gravitropic curvature of roots of maize and pea. Furthermore, the level of ethylene in the intercellular space of the roots was increased by submergence. These results suggest that the accumulation of ethylene in the tissue is at least partly involved in suppression of transmission of the gravity signal or of differential growth in maize and pea roots under conditions of submergence.Abbreviations AOA aminooxyacetic acid - 3-D three-dimensional Dedicated to Professor Andreas Sievers on the occasion of his retirementWe thank Professor H. Suge and Drs. H. Takahashi and H. Kataoka, Tohoku University and Dr. T. Suzuki, Yamagata University, for helpful suggestions. The present study was supported in part by a Grant for Basic Research in Space Station Utilization from the Institute of Space and Astronautical Science, Japan.     

Submergence-tolerant rice withstands complete submergence even in saline water: Probing through chlorophyll a fluorescence induction O-J-I-P transients

Plants experience multiple abiotic stresses during the same growing season. The implications of submergence with and without saline water on growth and survival were investigated using four contrasting rice cultivars, FR13A (submergence-tolerant, salinity-susceptible), IR42 (susceptible to salinity and submergence), and Rashpanjor and AC39416 (salinity-tolerant, submergence-susceptible). Though both FR13A and IR42 showed sensitivity to salinity, FR13A exhibited higher initial biomass as well as maintained greater dry mass under saline condition. Greater reduction of chlorophyll (Chl) contents due to salinity was observed in the susceptible cultivars, including FR13A, compared to the salinity-tolerant cultivars. Exposure of plants to salinity before submergence decreased the survival chance under submergence. Yet, survival percentage under submergence was greater in FR13A compared to other cultivars. Generally, the reduction in the Chl content and damage to PSII were higher under the submergence compared to salinity conditions. The submergence-tolerant cultivar, FR13A, maintained greater quantities of Chl during submergence compared to other cultivars. Quantification of the Chl a fluorescence transients (JIP-test) revealed large cultivar differences in the response of PSII to submergence in saline and nonsaline water. The submergence-tolerant cultivar maintained greater chloroplast structural integrity and functional ability irrespective of the quality of flooding water.     

Molecular Genetics of Submergence Tolerance in Rice: QTL Analysis of Key Traits

Flash flooding of young rice plants is a common problem forrice farmers in south and south-east Asia. It severely reducesgrain yield and increases the unpredictability of cropping.The inheritance and expression of traits associated with submergencestress tolerance at the seedling stage are physiologically andgenetically complex. We exploited naturally occurring differencesbetween certain rice lines in their tolerance to submergenceand used quantitative trait loci (QTL) mapping to improve understandingof the genetic and physiological basis of submergence tolerance.Three rice populations, each derived from a single cross betweentwo cultivars differing in their response to submergence, wereused to identify QTL associated with plant survival and variouslinked 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 underdifferent field conditions, locations and years. Several majorQTL determining plant survival, plant height, stimulation ofshoot elongation, visual tolerance score and leaf senescenceeach mapped to the same locus on chromosome 9. These QTL weredetected consistently in experiments across all years and inthe genetic backgrounds of all three mapping populations. SecondaryQTL influencing tolerance were also identified and located onchromosomes 1, 2, 5, 7, 10 and 11. These QTL were specific toparticular traits, environments, or genetic backgrounds. Allidentified QTL contributed to increased submergence tolerancethrough their effects on decreased underwater shoot elongationor increased maintenance of chlorophyll levels, or on both.These findings establish the foundations of a marker-assistedscheme for introducing submergence tolerance into agriculturallydesirable cultivars of rice.     

Molecular and Physiological Mechanisms of Flooding Avoidance and Tolerance in Rice

Submergence is one of the major constraints in rice production. The main factor limiting rice survival during submergence is oxygen deprivation. To cope with flooding conditions, rice has developed two survival strategies: either rapid elongation of the submerged tissues to keep up with the rising water level or no elongation to save carbohydrate resources for maintenance of energy production under submerged and concomitant hypoxic conditions. The survival strategies used by rice have been studied quite extensively and the role of several phytohormones in the elongation response has been established. The mechanisms of submergence tolerance include metabolic changes, for instance, the shift to an ethanolic fermentation pathway, reduced elongation growth to save carbohydrates and energy for maintenance processes, and protective antioxidant systems. Current molecular technology can provide tools for the understanding of mechanisms developed by rice to survive submergence. In addition, cloning of genes related to submergence tolerance might open new ways to genetic improvement of this crop.