Effects of salt stress on growth, mineral nutrition and proline accumulation in relation to osmotic adjustment in rice (Oryza sativa L.) cultivars differing in salinity resistance

Salt-resistant rice cultivars Nona Bokra and IR 4630 exposed at the seedling stage during one or two weeks to 0, 20, 30, 40 or 50 mM NaCl accumulated less Na, Cl, Zn and proline and more K at root and shoot levels than salt-sensitive I Kong Pao and IR 31785. Aiwu, a moderately resistant genotype, exhibited an intermediate behaviour. P transport from root to shoot was inhibited in the most sensitive cultivar IR 31785. Accumulation of Na and Cl and decrease in K content at the shoot level were restricted to the oldest leaves in salt-resistant genotypes while proline accumulated in the youngest leaves in all cultivars. In the presence of NaCl, the osmotic potentials of the roots and of the oldest and youngest leaves were lower in the salt-resistant than in the salt-sensitive genotypes, differences among genotypes increasing with stress intensity. Proline did not appear to be involved in osmotic adjustment in salt-stressed rice plants and the significance of its accumulation is discussed in relation to salinity resistance.Abbreviations cv(s). cultivar(s) - EC electrical conductivity - IKP I Kong Pao - J rate of ion transport - MCW methanol-chloroform-water - PAR photon flux density - Pc partitioning coefficient - RGR mean relative growth rate - RI resistance index - s osmotic potential     

Carbohydrate and osmotic requirements for high-frequency plant regeneration from protoplast-derived colonies of indica and japonica rice varieties

The effects were studied of various carbohydrates and osmoticstress, created by high agarose or carbohydrate concentrations,on the regeneration of fertile plants from protoplast-denvedcolonies of several indica (IR43, Jaya, Pusa Basmati 1) andjaponica (Taipei 309) rice varieties. Observations of the culturesdeveloped on media containing one of these carbohydrates (cellobiose,fructose, glucose, lactose, maltose, mannitol, sorbitol or sucrose),each at 88 mM, indicated that maltose was the preferential carbonsource for the proliferation of embryogenic callus and shootregeneration. Maltose-containing medium induced shoot formationin 24–66% of the protoplast-derived tissues, dependingupon the rice variety, compared to shoot regeneration from 4–32%of the tissues in sucrose-supplemented medium. Media containing288 mM maltose or an equimolar combination of 88 mM maltoseand 200 mM mannitol, caused water loss from calli and promotedthe growth of embryogenic calli. These calli formed shoots withgreater frequencies when subsequently transferred to shoot regenerationmedium with 88 mM maltose. A medium containing 88 mM maltoseand semi-solidified with 1.0% (w/v) instead of 0.5% (w/v) agarosehad a similar beneficial effect on the growth of embryogeniccalli and simultaneously supported high-frequency (48–55%)shoot formation. The optimum shoot regeneration frequencies(60–78%) were obtained when protoplast-derived colonieswere serially cultured on to shoot regeneration medium containing1.0% (w/v) agarose for 4 weeks, followed by a 2-week cultureperiod on the same medium with 0.5% (w/v) agarose. Plants regeneratedon medium containing maltose and/or 1.0% (w/v) agarose werephenotypically normal and fertile. Key words: Carbohydrates, Oryza sativa L, indica and japonica rice, osmotic stress, plant regeneration, protoplast-derived colonies     

Effects of nitrogen nutrition on salt-stressed Nicotiana tabacum var. Samsun in vitro plantlets

Tobacco shoots were grown in vitro for 35 d, in MS culture mediummodified to include various sources (nitrate-N, ammonium-N ora mixture) and levels (0–120 mM) of N, and in the presenceof 0–180 mM NaCI or iso-osmotic concentrations of mannitol.Growth of control plantlets was significantly inhibited whenNH₄⁺-N was the sole N source, and at high (120 mM) NO^–₃-N supply. Under conditions of salt stress (90 and 180 mM NaCI)growth was repressed, with roots being more severely affectedthan shoots. Salinity also inhibited root emergence in vitro.The only alleviation of the salt stress by nitrate nutritionobserved in this study was on shoot growth parameters of plantletsgrown on 60 mM NO^–₃-N and 90 mM NaCI. Although both weresignificantly inhibited by NaCI, nitrate reduc-tase activitywas more severely affected than nitrate uptake. When mannitolreplaced NaCI in the culture medium, similar Inhibition of growth,nutrient uptake and enzyme activity were recorded. These observations,together with the relatively low recorded values for Na⁺ andCI^– uptake, indicate that under in vitro salt stress conditionsthe negative effects of NaCI are primarily osmotic. Key words: Growth, nitrogen metabolism, osmotic stress, salinity     

Response of rice varieties to soil salinity and air humidity: A possible involvement of root-borne ABA

In a phytotron experiment four rice varieties (Pokkali, IR 28, IR 50, IR 31785-58-1-2-3-3) grown in individual pots were subjected to low (40/55% day/night) and high (75/90%) air humidity (RH), while soil salinity was gradually increased by injecting 0, 30, 60 or 120 mM NaCl solutions every two days. Bulk root and stem base water potential (SWP), abscisic acid (ABA) content of the xylem sap and stomatal resistance (rs) of the youngest fully expanded leaf were determined two days after each salt application. The SWP decreased and xylem ABA and rs increased throughout the 8 days of treatment. The effects were amplified by low RH. A chain of physiological events was hypothesized in which high soil electric conductivity (EC) reduces SWP, followed by release of root-borne ABA to the xylem and eventually resulting in stomatal closure. To explain varietal differences in stomatal reaction, supposed cause and effect variables were compared by linear regression. This revealed strong differences in physiological reactions to the RH and salt treatments among the test varieties. Under salt stress roots of IR 31785-58-1-2-3-3 produced much ABA under low RH, but no additional effect of low RH on rs could be found. By contrast, Pokkali produced little ABA, but rs was strongly affected by RH. RH did not affect the relationships EC vs. SWP and SWP vs. ABA in Pokkali, IR 28, and IR 50, but the relationship ABA vs. rs was strongly affected by RH. In IR 31785-58-1-2-3-3 RH strongly affected the relationship SWP vs. ABA, but had no effect on ABA vs. rs and EC vs. rs. The results are discussed regarding possible differences in varietal stomatal sensitivity to ABA and their implications for varietal salt tolerance.     

Robustness and Strategies of Adaptation among Farmer Varieties of African Rice (Oryza glaberrima) and Asian Rice (Oryza sativa) across West Africa

This study offers evidence of the robustness of farmer rice varieties (Oryza glaberrima and O. sativa) in West Africa. Our experiments in five West African countries showed that farmer varieties were tolerant of sub-optimal conditions, but employed a range of strategies to cope with stress. Varieties belonging to the species Oryza glaberrima – solely the product of farmer agency – were the most successful in adapting to a range of adverse conditions. Some of the farmer selections from within the indica and japonica subspecies of O. sativa also performed well in a range of conditions, but other farmer selections from within these two subspecies were mainly limited to more specific niches. The results contradict the rather common belief that farmer varieties are only of local value. Farmer varieties should be considered by breeding programmes and used (alongside improved varieties) in dissemination projects for rural food security.     

The ionic effects of NaCl on physiology and gene expression in rice genotypes differing in salt tolerance

The effects of ionic stress on the physiology and gene expression of two rice genotypes (IR4630 and IR15324) that differ in salt tolerance, were investigated by evaluating changes in the biomass, Na⁺ and K⁺ concentrations and applying the cDNA-AFLP technique to highlight changes in gene expression. Over 8 days of salinisation, the effect of NaCl on the reduction of biomass (dry weight) was apparent from 24 h after salinisation (the first time point), indicating that the consequences of the build up of Na⁺ (and Cl^-) in the leaves of both lines was rapid. Furthermore, root growth of IR15324 was much more sensitive to salt than that of IR4630 (the reduction in root dry weight compared to non-salinised plants was three times greater in IR15324 than IR4630). The two rice lines also differed in their Na⁺ accumulation in saline conditions, a difference that was more marked in the shoots, particularly at the final harvest, than in the roots. Under salt stress, the K⁺ content (µmol/shoot) increased over four successive harvests (24, 48, 96, 192 h) in both lines, but was always greater in IR4630 than in IR15324: differences in Na⁺/K⁺ ratio appear to be an important determinant of salt tolerance in rice. To separate osmotic from ionic effects of salt, mannitol was applied as a non-ionic osmoticum at an osmotic potential estimated to be equivalent to 50 mM NaCl. Messenger RNA was sampled at 0.5, 6, 24, 48 and 192 hours after salinisation. Several products (AFLP-bands) were detected, which were upregulated in the response to ionic effects of salt in the tolerant line (IR4630) and not expressed in the sensitive line (IR15324). Bioinformatic analysis indicated three of these AFLP-bands have a high-degree of sequence similarity with the genes encoding a proline rich protein, senescence associated protein and heat-shock protein. The data are novel in that they differentially highlight changes induced by the ionic rather than osmotic effects of salt and in a tolerant rather than a sensitive genotype. The possible roles of the products of these genes are discussed.     

Photothermal Responses of Flowering in Rice (Oryza sativa)

Durations from sowing to panicle emergence in 16 diverse genotypesof rice (Oryza sativa L.) were recorded in 13 different photothermalregimes, comprising constant and diurnally alternating temperaturesbetween 16 and 32 °C and photoperiods between 10.5 and 15.0h d^–1—all provided by controlled-environment growthcabinets. In 11.5 h days and at sub-optimal temperatures, relationsbetween the rate of progress towards panicle emergence and meantemperature were linear in all genotypes, and amongst thesethe base temperature at that photoperiod varied between 6.6and 11.9 °C. In most cases progress was most rapid at 24–26°C, i.e. the optimum temperature was much cooler than expectedfrom previously published values of times to panicle emergencein a less extensive range of photothermal regimes. Only in threecultivars was it warmer than 28 °C, and in these there weresufficient data to establish that relations between rates ofprogress to panicle emergence and photoperiod in the diurnallyalternating temperature regime of 28–20 °C are alsolinear. Also, the responses of these three cultivars provideno evidence of any interaction between the effects of photoperiodand temperature. We conclude, then, that the model in whichrate of development is a linear function of both temperatureand photoperiod with no interaction, which has been shown tobe common to many other species, also applies to rice. Differencesamong genotypes in relative sensitivity of rate of progresstowards panicle emergence to both temperature and to photoperiodwere considerable; japonica cultivars tended to be more sensitiveto temperature and less sensitive to photoperiod than indicacultivars. Four indica cultivars bred and selected at The InternationalRice Research Institute (IRRI) in the Philippines did not differ(P > 0.10) in their relations between rate of progress towardspanicle emergence and sub-optimal temperatures in a daylengthof 11.5 h, but the optimum temperature for cv. IR 36 was appreciablywarmer than that for the cvs IR 5, IR 8 and IR 42. Oryza sativa, rice, flowering, temperature, photoperiod, photothermal responses     

Heteroses in Net Photosynthetic Rate, Leaf Area, Tillering, and Some Physiological Characters of 35 F1 Rice Hybrids

Net photosynthetic rate, leaf area, tillering, shoot weight,and leaf area ratio for 35 F₁ rice (Oryza sativa L.) hybridsduring the vegetative stage were compared with correspondingvalues for parental varieties. Shoot weight and leaf area showedheterosis. However, hybrid vigour was not reflected in net photosyntheticrate. Leaf area was closely correlated with heteroses in shootdry weight and tillering. Thus F₁ rice hybrids grow vigorouslybecause of their high leaf area development, which is causedby high tillering. Key words: F₁ rice hybrid, Net photosynthetic rate, Leaf area     

Abscisic Acid Accumulation in Detached Leaves of Rice (Oryza sativa L.) in Response to Water Stress: A Correlation with Leaf Size

When water stress was imposed on detached leaves of two rice(Oryza sativa L.) cultivars, more ABA per unit fresh weightaccumulated in IR20, a small-leaved cultivar, than in 63–83,a large-leaved cultivar; the difference being up to threefold.In an F₂ population of a cross between the two cultivars ABAaccumulation was found to be significantly negatively correlatedwith leaf fresh weight. This correlation persisted in the F₃generation. Such a correlation was not evident, however, whena number of rice cultivars, which varied widely in leaf size,were examined. The difference in ABA accumulation between IR20 and 63–83was not accounted for by different spatial patterns of waterloss or ABA accumulation within a leaf, and cultivar differencesin ABA content were maintained both across, and at various positionsalong the leaf. No major differences in leaf anatomy were observed between thetwo cultivars. Differences found in leaf water relations characteristicswere few and generally minor. It therefore seems unlikely thatthese properties account for the difference between the cultivarsin the ability to accumulate ABA or for the correlation withleaf size. Oryza sativa L, rice, water stress, abscisic acid, leaf size     

Photosynthesis and Transpiration in Rice as Influenced by Soil Moisture and Air Humidity

The paper describes the effect of soil moisture content andair humidity on CO₂ exchange (P_N), CO₂ diffusion resistance(Cr) and transpiration (E) in four varieties of japonica rice(Oryza sativa L.). A decrease in soil moisture content reducedthe rate of photosynthesis to a varying degree in the varieties.Reduction in photosynthesis was attributed to increase in Cr.The effect of low soil moisture on photosynthesis and CO₂ diffusionwas further intensified by decrease in air humidity. By maintaininga high humidity in the air around the leaves however, the effectof soil moisture deficiency was reduced considerably, exceptin Rikuto Norin 21 which was very sensitive to soil-moisturedeficiency alone. Dryness of the air enhanced the transpirationrate, although the increase was relatively less in the plantsfacing a simultaneous water crisis at the root surface. In plantsgrowing under flooded conditions, a decrease in air humiditycaused a slight depression in P_N despite the simultaneous decreasein Cr. Oryza sativa L., rice, photosynthesis, transpiration, diffusion resistance, soil moisture, air humidity     

Endogenous Gibberellins in Aeginetia indica,a Parasitic Plant,and Its Host,Miscanthus sinensis

Endogenous gibberellins in a parasitic plant, Aeginetia indica L., and its host, Miscanthus sinensis Andress (eulalia) were analyzed. Gibberellins of the early-non-hydroxylation pathway and their putative metabolites were identified as the major endogenous gibberellins from both types of A. indica parasitizing M. sinensis and parasitizing Oryza sativa L. (rice). Members of both the early-non- and early-13-hydroxylation pathways were detected in the host M. sinensis. Since the early-13-hydroxylation pathway has been reported to be the major pathway operating in vegetative tissues of O. sativa, these results suggest that A. indica can biosynthesize gibberellins independent of its hosts.     

Durations of the Photoperiod-sensitive and Photoperiod-insensitive Phases of Development to Flowering in Four Cultivars of Rice (Oryza sativa L.)

The durations of the photoperiod-sensitive and photoperiod-insensitivephases of development to panicle emergence were estimated infour contrasting indica cultivars of rice (Oryza sativa L.)in a reciprocal-transfer experiment. Plants were grown in potsin glasshouses maintained at warmer (32/26 C) or cooler (28/20C) day/night temperatures, and the durations from sowing topanicle emergence were determined for plants moved from relativelyshort (11 h) to relatively long (13.5 h) days and vice versaat various times after sowing. Panicle emergence was delayedby long days in all cultivars, but the traditional cvs Carreonand Peta were much more sensitive to photoperiod than the moderncvs IR8 and IR36 The durations of the photoperiod-insensitivepre-inductive phase (equivalent to some definitions of the basicvegetative phase) varied from 14.4 d in cv. Carreon at 32/26C to 42.0 d in cv. IR8 at 28/20 C. In all cultivars this initialphase was of a longer duration in the cool than in the warmregime. The duration of the photoperiod-insensitive post-inductivephase was also consistently greater, but usually only slightso, at cool than relatively warm temperatures; it varied from6.8 d in cv. IR8 at 32/26 C to 272 d in cv. Carreon at 28/20C. As expected, the length of the intervening photoperiod-sensitiveinductive phase was greater in long days, but the effect oftemperature on these durations was not consistent; for example,these durations were longer in warm than in cool temperaturesin cv. 1R8 but, if anything, they were slightly longer in coolthan in warm temperatures in cv. IR36. This difference is compatiblewith previous findings that cv. IR36 has a warmer optimum temperaturefor rate of progress towards panicle emergence than cv. IR8.A subsequent reciprocal-transfer experiment with cv. Peta providedestimates of the durations of the photoperiod-insensitive andphotoperiod-sensitive phases of pre-flowering development whichwere compatible with our earlier estimates. Furthermore, panicleinitiation was found to occur after about 80% of the photoperiod-sensitiveinductive phase had elapsed. We conclude that although the durationof the photoperiod-insensitive pre-inductive phase in rice isgreater than in many other annual crops, genotypic variationin this duration may well be less than was previously deduced.We also conclude that, despite common assumptions to the contrary,photoperiod-sensitivity during rice plant development does notend at panicle initiation. Oryza sativa L., rice, panicle initiation, panicle emergence, photoperiodism, temperature     

Biochemical Basis of Different Sensitivity to Methotrexate in Daucus carota and Oryza sativa Cell Cultures

The biochemical basis of the different sensitivity to methotrexateof Daucus carota and Oryza sativa cell cultures has been investigated.Carrot cells have a dihydrofolate reductase activity about tentimes higher than rice cells. In addition, they show a loweruptake rate of the inhibitor. No relevant differences have beenfound in the K_m value for the dihydrofolate of the two enzymesand in the degree of inhibition of their activity by methotrexate. Key words: Dihydrofolate reductase, Methotrexate resistance, Plant cell suspension cultures, Oryza sativa, Daucus carota     

Effects of UVB Radiation on Stomatal Density and Opening in Rice (Oryza sativa L.)

Ultraviolet-B (UVB, wavelength 280-320 nm) radiation has beendemonstrated to affect growth and development of many plants.This study was conducted to determine the effect of UVB radiationon stomatal density and opening of Oryza sativa and to testif the stomatal response to UVB was associated with differentsensitivity of growth to UVB in four cultivars. Ten-day-oldseedlings of IR45 and IR74 (UVB sensitive), and IR64 and IR30(UVB less sensitive), were subjected to UVB radiation in a glasshousefor 6 h d^-1 for 4 weeks. The unweighted UVB radiation was 1·94W m^-2 for UVB treatment and 0·15 W m^-2 for control. Leafarea and plant dry mass were determined every 2 weeks whilestomatal density and opening were recorded weekly. Results showedthat a 2-week UVB treatment had no effect on the leaf area orplant dry mass of any test cultivar, but significantly reducedstomatal density and opening in IR45 and IR74. Under 4-weekUVB exposure, leaf area and plant dry mass of IR45 and IR74were significantly reduced. Stomatal density decreased in allcultivars, except in IR64. Greater reduction of stomata on theadaxial surface than on the abaxial surface under 3 and 4 weeksof UVB exposure suggests a direct effect of UVB radiation onstomata. IR45 and IR74 showed significant reductions in stomatalopening after 2 weeks of exposure to UVB, while stomatal openingin IR30 and IR64 decreased significantly after only 4 weeksof UVB treatment. Difference in plant dry mass between UVB treatedand control plants was significantly correlated with the reductionsin stomatal opening and density on adaxial surface under UVBtreatment. Thus, reduction in dry mass of rice plants underUVB in the glasshouse could be attributed to decrease in stomataldensity and opening.Copyright 1995, 1999 Academic Press Oryza sativa, UVB radiation, stomatal density, stomatal opening     

Cytochemical localization of ATPase plasma membrane and acid phosphatase by cerium-based method in a salt-adapted cell line of Pisum sativum

In order to study a possible application of cerium-based techniquesin plant cells, ATPase and acid phosphatase activities havebeen compared in two cell lines of Pisum sativum calli, onesensitive to NaCI and the other selected to be grown under salinity(85 mM NaCI). ATPase activity was unchanged and localized inthe plasma membrane of both cell lines. Acid phosphatase activitywas significantly increased in the salt-selected line and localizedin the cell walls, Golgi complex, multivesicular bodies andvacuoles. These results indicated a possible involvement ofboth activities in the maintenance of cell growth in the selectedline under saline conditions. Key words: Acid phosphatase, Pisum sativum, plasma membrane ATPase, salt stress, ultrastructure     

Polyamines as modulators of salt tolerance in rice cultivars

The effect of NaCl on the endogenous levels of diamine, putrescine and polyamines, spermidine and spermine, was studied in the shoot system of salt-tolerant and salt-sensitive lines of rice (Oryza sativa L.) cultivars during three growth stages. Salt stress increased the levels of diamine and polyamine in varying degrees among nine rice cultivars investigated. Salt tolerant AU1, Co43, and CSC1 were effective in maintaining high concentrations of spermidine and spermine, while the content of putrescine was not significantly altered in all the growth stages when plants were exposed to salinity. The salt sensitivity in rice was associated with excessive accumulation of putrescine and with low levels of spermidine and spermine in the shoot system of salt-sensitive cultivars Co36, CSC2, GR3, IR20, TKM4, and TKM9 under saline condition. One of the possible mechanisms of saline resistance was observed to be due to the highly increased polyamines against the low increase in diamines. Alternatively, the salt sensitivity could be due to high increase of diamines and an incapacity to maintain high levels of polyamines.     

RICD: A rice <Emphasis Type="Italic">indica</Emphasis>cDNA database resource for rice functional genomics

Background  

The Oryza sativa L. indica subspecies is the most widely cultivated rice. During the last few years, we have collected over 20,000 putative full-length cDNAs and over 40,000 ESTs isolated from various cDNA libraries of two indica varieties Guangluai 4 and Minghui 63. A database of the rice indica cDNAs was therefore built to provide a comprehensive web data source for searching and retrieving the indica cDNA clones.     

Salinity-induced enhancement of L-myo-inositol 1-phosphate synthase in rice (Oryza sativa L.)

The salt-tolerant varieties of rice (Oryza sativa L.) exhibit enhanced activity of the chloroplast form of L-myo-inositol 1-phosphate synthase (EC 5.5.4.1) under NaCl treatment either during the seedling stage or in fully grown plants during field growth. The salt-induced enhancement was noticeable only in chloroplasts from light-grown plants. The effects of these treatments on the cytosolic inositol synthase activity were less pronounced. While the effect of salt on the activity of the two forms was marginal in the salt-sensitive varieties during seedling growth, salinity affected the chloroplast inositol synthase activity adversely in these varieties during growth of the plants under field conditions. The salt-enhanced activities of inositol synthase(s) in the highly salt-tolerant varieties studied were found to be comparable to that observed in Porteresia coarctata, a halophytic wild rice species. The implications of these findings, which suggest a role of the inositol pathway in osmoregulation, are discussed.     

Cultivar Differences in Growth and Chloroplast Ultrastructure in Rice as Affected by Nitrogen

Plant growth, leaf protein and chlorophyll content, and chloroplastultrastructure as affected by nitrogen (N) were examined infour rice (Oryza sativa L.) cultivars grown in culture solutionunder controlled conditions. Increasing N concentration generallyincreased height and shoot dry weight of all cultivars. Cultivardifferences were significant at normal N level (40 ppm). Amongcultivars, IR8 was most responsive to increasing N, having thesignificantly highest shoot dry weight and protein content.Total chlorophyll and protein contents varied with cultivarand N, but chlorophyll a/b ratio remained constant. At the ultrastructurallevel, chloroplasts had generally well-developed grana and stromalamellae at 40 ppm.N. Chloroplasts at high N had from one tofour times as many grana as the N-deficient chloroplasts. Nitrogendeficiency reduced the size of the chloroplast, grana-stromalamellae and resulted in fewer poorly stacked grana. Increasingthe N level (120 ppm) above the normal level did not significantlyaffect chloroplast size of any cultivar, except for IR8 whichhad the largest chloroplast. A reduction in the number of starchgrains was observed in IR8, but more were present in ER36 underN-deficient conditions. The size of starch grains was not affectedby N and did not differ among cultivars. Plastoglobuli appearedto be larger under N-deficient conditions. Nitrogen had no effecton the number of plastoglobuli but cultivar differences existed.The highly N-responsive IR8 (based on dry weight) had the largestchloroplast which increased with N level. The increase in chloroplastsize accounted for the increase in both chlorophyll and proteincontents and, consequently, dry weight. Key words: Oryza sativa L., chloroplast, chlorophyll, protein