Ethylene and the growth of rice seedlings

Etiolated whole rice seedlings enclosed in sealed vials produced ethylene at a rate of 0.9 picomole per hour per seedling. When 2-centimeter-long shoots were subdivided into 5-millimeter-long sections, the sections containing the tip of the shoot evolved 37% of the total ethylene with the remaining 63% being produced along a gradient decreasing to the base of the shoot. The tip of the coleoptile also had the highest level of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid and of the ethylene-forming enzyme activity. Ethylene is one of the factors controlling coleoptile elongation. Decapitation of the seedling reduced ethylene evolution to one-third its original level and inhibited coleoptile growth. In short-term experiments, the growth rate of decapitated seedlings was restored to almost that of intact seedlings by application of ethylene at a concentration of 10 microliters per liter. Apart from ethylene, O₂ also participates in the control of coleoptile growth. When rice seedlings were grown in a gas mixture of N₂ and O₂, the length of the coleoptiles reached a maximum at a concentration of 2.5% O₂. Lower and higher concentrations of O₂ reduced coleoptile growth. The effect of exogenous ethylene on coleoptile growth was also O₂ dependent.     

Effects of ethanol on growth of rice seedlings

Effects of ethanol, the end product of ethanolic fermentation, on the growth of rice (Oryza sativa L.) seedlings were determined as a means of evaluating growth responses under anoxia. The ethanol concentrations in roots and coleoptiles of the seedlings subjected to 48 h-anoxia, and in their culture medium were 23 and 32 µmol g^–1 fresh weight, and 19 µmol ml^–1, respectively. The growth of the roots and coleoptiles of the seedlings was restricted by exogenous ethanol at concentrations above 50 mM and 100 mM, respectively, suggesting that the roots are more sensitive to ethanol than the coleoptiles.     

Levels of endogenous polyamines and NaCl-inhibited growth of rice seedlings

The role of endogenous polyamines in the control of NaCl-inhibited growth of rice seedlings was investigated. Putrescine, spermidine and spermine were all present in shoots and roots of rice seedlings. NaCl treatment did not affect spermine levels in shoots and roots. Spermidine levels in shoots and roots were increased with increasing concentrations of applied NaCl. NaCl at a concentration of 50 mM, which caused only slight growth inhibition, drastically lowered the level of putrescine in shoots and roots. Addition of precursors of putrescine biosynthesis (L-arginine and L-ornithine) resulted in an increase in putrescine levels in NaCl-treated shoots and roots, but did not allow recovery of the growth inhibition of rice seedlings induced by NaCl. Pretreatment of rice seeds with putrescine caused an increase in putrescine level in shoots, but could not alleviate the inhibition effect of NaCl on seedling growth. The current results suggest that endogenous polyamines may not play a significant role in the control of NaCl-inhibited growth of rice seedlings.Abbreviations PUT putrescine - SPD spermidine - SPM spermine     

Regulation of growth in stem sections of deep-water rice

Submergence in water greatly stimulates internodal elongation in excised stem sections of deep-water rice (Oryza sativa L. cv. Habiganj Aman II) and inhibits growth of leaf blades and leaf sheaths. The highest rates of internodal growth have been observed in continuous light. Very little growth occurs in submerged sections kept in darkness or incubated under N₂ in the light. The effect of submergence on the growth of deep-water rice is, at least in part, mediated by C₂H₄, which accumulates in the air spaces of submerged sections. This accumulation results from increased C₂H₄ synthesis in the internodes of submerged sections and reduced diffusion of C₂H₄ from the tissue into the water. Increased C₂H₄ levels accelerate internodal elongation and inhibit the growth of leaves. Compounds capable of changing the rate of C₂H₄ synthesis, namely aminoethoxyvinylglycine, an inhibitor of C₂H₄ synthesis, and 1-aminocyclopropane-1-carboxylic acid, the immediate, precursor of C₂H₄, have opposite effects on growth of internodes and leaves. The enhancement of internodal elongation by C₂H₄ is particularly pronounced in an atmosphere of high CO₂ and low O₂. The increase in C₂H₄ synthesis in internodes of submerged sections is primarily triggered by reduced atmospheric concentrations of O₂. The rate of C₂H₄ evolution by internodes isolated from stem sections and incubated in an atmosphere of low O₂ is up to four times greater than that of isolated internodes incubated in air. In contrast, C₂H₄ evolution from the leaves is reduced under hypoxic conditions. The effect of submergence on growth of stem sections of deep-water rice can be mimicked by exposing non-submerged sections to a gas mixture which is similar to the gaseous atmosphere in the internodal lacunae of submerged sections, namely 3% O₂, 6% CO₂, 91% N₂ (by vol.) and 1 l l^-1 C₂H₄. Our results indicate that growth responses obtained with isolated rice stem sections are similar to those of intact deep-water rice plants.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine     

Biological activity of pestalotins on the elongation growth of rice seedlings

The effect of (–)-pestalotin, (+)-pestalotin and (+)-epipestalotinon the elongation growth of rice seedlings was studied. Thesecompounds did not influence elongation when given alone, although(–)-pestalotin and (+)-epipestalotin synergistically enhanced GA₃-induced elongation. (Received May 20, 1977; )     

Aluminum-inhibited root growth of rice seedlings is mediated through putrescine accumulation

The effects of AlCl₃ on growth and polyamine levels of rice roots were investigated. When rice roots were treated with AlCl₃, root growth was markedly inhibited. AlCl₃ treatment resulted in a higher putrescine content and lower spermidine and spermine contents in rice roots. d-Argnine and α-methylornithine, inhibitors of putrescine biosynthesis, caused a reduced content of putrescine in rice roots under Al stress. AlCl₃ treatment also resulted in a decrease in diamine oxidase activity in rice roots. The growth of rice roots in the presence of AlCl₃ was recovered after the addition of d-arginine or α-methylornithine. The protective effect of d-arginine or α-methylornithine in counteracting AlCl₃-inhibited growth of rice roots is unlikely caused by reduction of Al uptake. Furthermore, the effect of the growth recovery in AlCl₃-treated rice roots by d-arginine or α-methylornithine was reversed by the addition of putrescine. Our results strongly suggest that putrescine accumulation is a factor causing growth inhibition of rice roots under Al tress. Evidence is also presented to show that lignification is responsible for putrescine- and AlCl₃-inhibited growth of rice roots.     

Aluminum uptake and aluminum-induced rapid root growth inhibition of rice seedlings

Aluminum (Al) inhibits root growth in acidic soil, but the site of action of Al remains unclear. We investigated whether the rate of Al accumulation correlates to Al-indeced rapid root growth inhibition in rice seedlings (Oryza sativa L. cv. Youngnam). Growth of roots was significantly inhibited by 100 μM AICI₃, as early as 1 h after the treatment. The inhibition of root growth was strongly dependent on Al concentration (l₅₀ = 20 (μM) and Al-exposure time (l₅₀ = 23 min at 25 μM Al) in a solution of 10 mM KCI and 1 mM CaCl₂ buffered by 10 mM Mes/KOH (pH 4.5). Using ICPES, massive uptake of Al by roots was observed even at 15 min treatment of 25 μM Al. The kinetics of Al uptake by the roots closely corresponded to the inhibitory effects of Al on root growth. When the roots of seedlings were exposed to 50 (μM Al for 1 h, then sectioned and stained with hematoxylin, all cell types of the roots showed the presence of Al in the cytoplasm. These results indicate that Al was rapidly taken up into the root cells and thereby reduced root growth.     

Synergistic action of ethylene with helminthosporic acid in the growth of rice seedlings

Ethylene was found to promote the growth of rice seedlings synergisticallywith helminthosporic acid. (Received July 10, 1973; )     

Significant contribution of boron stored in seeds to initial growth of rice seedlings

Rice plants are relatively insensitive to low boron environment. We examined the role of boron stored in seeds in growth of rice seedlings in regard to its insensitivity to low boron condition. In experiments using seeds without the removal of the endosperm, the effect of low boron treatments (0.18 and 0.03 μM B) was not evident on growth of seedlings until 5 weeks from germination. Analysis of boron distribution among different organs in rice grain demonstrated that the endosperm contained 92% of total boron in brown rice and husks contained twice as much boron as the brown rice. To examine the role of boron stored in the endosperm, endosperm were removed 5 days after de-husking and imbibition and the plants were subjected to boron treatments. The growth of the plants without endosperm was much reduced after 10 days under the 0.03 μM boron compared with the plants grown with the normal level (18 μM B). With the normal level of boron supply, the growth did not differ between the plants with and without endosperm, suggesting that growth difference between seedlings with and without endosperm under low B condition is not due to lack of starch or proteins, but due to lack of boron stored in seeds. Tracer experiments confirmed that a substantial amount of B was supplied from the seed to the seedling and that B limitation increased the B supply from seeds to seedlings, especially to roots. Our results established that boron stored in seeds, principally in the endosperm, contributes significantly to the growth of rice seedlings when external boron supply is limited.     

Effect of extracellular products ofAulosira fertilissima on the growth of rice seedlings

Summary The effect of extracellular products fromAulosira fertilissima on the growth of rice seedlings (IR-8) has been studied. There is indication of liberation of a root-promoting hormone from the alga. The growth pattern of rice seedlings, treated with algal filtrate, parallels that of seedlings treated with gibberellic acid. An increased growth of rice seedlings results when they are treated with algal filtrate obtained from the exponential phase of the alga. The amino acid-treated seedlings do not show any marked increase in growth. Filtrate fromAnacystis nidulans, on the other hand, do not show any growth promoting response. The possibility of liberation of kinins byA. fertilissima does not appear to be involved.     

Selection of RAPD marker for growth of seedlings at low temperature in rice.

We have developed a polymerase chain reaction (PCR)-based assay that could effectively reduce the time period required to screen and select the cold tolerance gene of rice seedlings under field conditions. The two specific random amplified polymorphic DNA (RAPD) fragments for the assay were identified on the basis of quantitative trait loci (QTL) analysis which were found to be tightly linked to cold sensitivity. The two RAPD fragments, OPT8(600) in the cold sensitivity rice cultivar 'Dular (indica)' and OPU20(1200) in the resistance rice cultivar 'Toyohatamochi (japonica)', were identified after screening 11 RAPD fragments using 2 random primers on the genomic DNAs of 'Dular' and 'Toyohatamochi'. These primers, when used in a multiplexed PCR, specifically amplified a 0.6 kb and a 1.2 kb fragment in the sensitive and resistant rice cultivars, respectively. When this assay was performed on the genomic DNAs of 16 japonica, 3 Tongil (indica/ japonica), and 2 indica rice cultivars, the primers amplified a 0.6 kb fragment in all of the cold sensitivity rice cultivars or 1.2 kb fragment in all of the resistance ones. These markers can be of potential use in the marker-assisted selection (MAS) for cold tolerance in rice seedling. As screening for resistance can now be conducted independent of the availability of low temperature, the breeding of cold tolerance cultivars can be hastened.     

Suppression and promotion of growth by ethylene in rice seedlings depends on ambient humidity

We examined the effect of ethylene on the growth of rice seedlings (Oryza sativa L.) at various degrees of humidity. Ethylene significantly suppressed the growth of shoots when applied to seedlings grown under 30% relative humidity (RH), but promoted the growth of shoots when applied to seedlings grown under 100% RH. The application of gibberellic acid (GA₃) promoted the elongation of shoots in seedlings grown under 30% and 100% RH. Ethylene inhibited the shoot elongation induced by GA₃ at 30% RH, but enhanced the elongation induced by GA₃ at 100% RH. These results indicate that ethylene can either promote or suppress the growth of rice shoots depending on ambient humidity, and that these actions of ethylene may be mediated through modulating the responsiveness of shoots to gibberellin.     

Similarities and differences between phytochrome-mediated growth inhibition of coleoptiles and seminal roots in rice seedlings

In rice, light is known to inhibit the growth of coleoptiles and seminal roots of seedlings through phytochrome. Here we investigated the light-induced growth inhibition of seminal roots and compared the results with those recently determined for coleoptiles. Although three rice phytochromes, phyA, phyB and phyC functioned in a similar manner in coleoptile and seminal root, the Bunsen-Roscoe law of reciprocity was not observed in the growth inhibition of seminal root. We also found coiling of the seminal root at the root tip which appeared to be associated with the photoinhibition of seminal root growth. This could be a new light-induced phenomenon in certain cultivars of rice.Key words: growth, hypocotyl, Oryza sativa, phytochrome, seminal rootPhytochrome-mediated growth inhibition was reported for both coleoptiles and seminal roots of rice seedlings in the same year by two research groups in Nagoya and Tohoku University in Japan, respectively.^1,2 Forty years after the findings, a detailed photobiological study was carried out for the coleoptile growth inhibition.³ In this study, we examined photoinhibition of seminal root growth, and found similarities and differences between light-induced growth inhibition of the two organs in rice seedlings. Although coleoptile growth was inhibited by pulses of light, growth inhibition of seminal roots required light irradiation longer than 6 h. The Bunsen-Roscoe law of reciprocity was not observed in the growth inhibition of seminal root. Action spectra were determined for the growth inhibition of coleoptiles, and the mode of inhibition was found to depend on the age of the coleoptiles. At the early stage of development [40 h after inducing germination (AIG)], photoinhibition was predominantly due to the phyB-mediated low-fluence response (LFR), but at the late developmental stage (80 h AIG), it consisted of the phyA-mediated very low-fluence response (VLFR) as well as the phyB-mediated LFR.^3,4 In the case of root growth, the sensitivity of photoinhibition also depended on age, and was most sensitive in the period of 48–96 h AIG when seedlings were irradiated for 24 h. Using rice phytochrome mutants,⁵ we found that far-red light for root growth inhibition was perceived exclusively by phyA, that red light was perceived by both phyA and phyB, and that phyC had little or no role in growth inhibition. Furthermore, the fluence rate required for phyB-mediated inhibition was more than 10,000-fold greater than that required for phyA-mediated inhibition. These characteristics of photoinhibition in seminal roots are similar to those found in coleoptiles at the late stage of development.³ In seminal roots, photoinhibition appeared to be mediated by photoreceptors in the root itself.Interestingly, coiling of the root tips always occurred when root growth was inhibited under the light condition (Fig. 1B). Under continuous light irradiation, rice seeds germinated ∼30 h AIG. Seminal roots formed a coil at the root tips during the 48–96 h period AIG, and stopped growing. When they were irradiated for only 24 h on the 3rd day AIG, coils started to form just after the end of irradiation. The roots continued to coil for ∼28 h and then began growing straight again (Fig. 1C). The coils were larger and looser than those formed under continuous light condition (Fig. 1, Open in a separate windowFigure 1Light irradiation induces coiling of root tips in rice seedlings (Oryza sativa cv. Nipponbare). A rice seedling was grown in the dark (A), or in continuous white light (55 µole m⁻² s⁻¹) (B) for 7 d at 28°C. In (C), it was irradiated by white light for 24 h during the 48–72 h period after inducing germination, and kept in the dark again until the 7th day. Arrows and arrowheads indicate the seminal and crown roots, respectively. Seedlings were grown in glass tubes of 3-cm diameter.

Table 1

The size of coil of root tips formed after white light irradiation

Effect of algal extracts of Phormidium species on growth and development of rice seedlings

Summary The effect of growth-promoting substances in extracts of Phormidium foveolarum, P. tenue, and P. frigidum on rice seedlings has been studied by presoaking seed treatment. The observations show that the growth and development of roots and shoots is markedly stimulated, in seeds treated with P. foveolarum and P. tenue and the results are highly significant statistically.     

Inhibitory effects of monoamine oxidase inhibitors on the growth of pea and rice seedlings

Two monoamine oxidase inhibitors of the hydrazine-type, safrazineand nialamide, inhibited growth in seedlings of rice and pea.We demonstrated histochemically that monoamine oxidase is locatedchiefly in sieve tubes and in the epidermis of pea seedling.Activity of this enzyme was high in the apical part of the epicotyl,decreasing toward the base. Inhibition of pea monoamine oxidaseby safrazine and nialamide was observed histochemically andwith an extract from the epicotyl. This supports the hypothesisthat indole-3-acetic acid (IAA) is formed from tryptamine byamine oxidase and that inhibition of this enzyme causes loweringof the auxin level, resulting in growth inhibition. Inhibitionof growth in rice seedlings by safrazine was reversed by theaddition of IAA to the culture medium. (Received May 6, 1970; )     

Water-stressed maize, barley and rice seedlings show species diversity in mechanisms of leaf growth inhibition

The possible occurrence of species diversity in mechanisms underlying leaf-growth inhibition by water stress, was investigated in related cereal plants. Water stress was generated by additions of the osmoticum polyethylene glycol 6000 to the root medium. Effects of external water potentials ranging from 0 to -0.6MPa, on early growth parameters of emerging leaves were measured under controlled environment conditions, using pairs of maize, barley or rice genotypes with differing resistance to water stress under field conditions. Water potentials of -0.4 MPa for 24 h, similarly reduced leaf growth, comparative production rates of leaf epidermal cells and cell size in all genotypes. These reductions did not appear to be caused by reductions in the osmotic potential gradients between the expanding leaf cells and their external water source. However, growth inhibition in maize and barley, was accompanied by significant reductions in comparative leaf and cell wall extensibility. Moreover, regression plots revealed good linear correlations (r=0.83** for maize and r=0.77** for barley) between the reductions in leaf growth induced by a series of water potentials and associated reductions in leaf extensibility. In contrast, the reduction in growth of rice leaves, was not accompanied by any significant changes in leaf or cell wall extensibility. Similarly, regression plots revealed poor correlations between leaf growth and leaf extensibility in both paddy and upland rice (r=0.17 and r=0.07, respectively). Thus, despite numerous inter-species similarities, biophysical changes associated with stress-induced leaf growth inhibition in maize and barley, differed from those in rice.Key words: Cell walls, extensibility, water stress, cereal diversity, leaf growth.      

Regulation of expansin gene expression affects growth and development in transgenic rice plants

To investigate the in vivo functions of expansins, we generated transgenic rice plants that express sense and antisense constructs of the expansin gene OsEXP4. In adult plants with constitutive OsEXP4 expression, 12% of overexpressors were taller and 88% were shorter than the average control plants, and most overexpressors developed at least two additional leaves. Antisense plants were shorter and flowered earlier than the average control plants. In transgenic plants with inducible OsEXP4 expression, we observed a close correlation between OsEXP4 protein levels and seedling growth. Coleoptile and mesocotyl length increased by up to 31 and 97%, respectively, in overexpressors, whereas in antisense seedlings, they decreased by up to 28 and 43%, respectively. The change in seedling growth resulted from corresponding changes in cell size, which in turn appeared to be a function of altered cell wall extensibility. Our results support the hypothesis that expansins are involved in enhancing growth by mediating cell wall loosening.     

Variation in the nitrate reductase of rice seedlings

Summary Nitrate reductase was induced in rice seedlings by nitrate and by chloramphenicol. During the induction period the different enzyme activities associated with nitrate reductase increased to different degrees. Nitrate induced high NADH-nitrate reductase activity and a great increase in the NADH-cytochrome c reductase activity which was associated with the nitrate reductase in a sucrose gradient. Chloramphenicol induced a nitrate reductase which had higher activity with NADPH than NADH. Chloramphenicol also induced a marked increase in NADPH-cytochrome c reductase activity as well as in NADH-cytochrome c reductase activity. Both activities were associated with the nitrate reductase in a sucrose gradient.After partial purification by sucrose gradient sedimentation or by starch gel electrophoresis, the nitrate reductase of rice induced by nitrate and chloramphenicol showed the same preference in pyridine nucleotide cofactors as was shown by the crude enzyme extracts.