Thigmomorphogenesis: The induction of callose formation and ethylene evolution by mechanical perturbation in bean stems

Mechanical perturbation by rubbing of the first internode of 11–12 day old plants of Phaseolus vulgaris L. cv. Cherokee wax induces the rapid deposition of callose in the cells of phloem and other tissues. Callose deposition begins immediately after mechanical perturbation, and shows a minor transient peak 1.5 h, and a major peak 6 h later. The callose gradually disappears and is gone after 3 days. If the stems are perturbed every day, the amount of callose decreases by day 2 but then gradually increases again through day 12. Both the top and bottom of the internode produce callose in response to mechanical perturbation. The evolution of ethylene in response to mechanical perturbation begins after 1 h, peaks at 2–3 h and is gone by 5–6 h. A spray of 10⁻² M 2-deoxy-D-glucose (DDG) completely blocks stem thickening, callose deposition and ethylene evolution due to mechanical perturbation. DDG at 10⁻⁵ to 10⁻⁴ M blocks callose production in mechanically perturbed stem segments and increases ethylene evolution from unperturbed stem segments to greater levels than those obtained by mechanically perturbed segments. It is concluded that mechanical perturbation of bean stems tissue induces deposition of callose more rapidly than it induces evolution of ethylene and that DDG can block both processes.     

Thigmomorphogenesis: membrane lipid and protein changes in bean plants as affected by mechanical perturbation and Ethrel

When bean plants (Phaseolus vulgaris L. cv. Red Cherokee Bush) are mechanically perturbed by rubbing, their stem elongation is inhibited and the stem thickness increases. The decrease in cell elongation and the increase in lateral cell divisions, which are partially responsible for this syndrome, were correlated with a decrease in the tree fatty acids and in the phospholipids of the membranes of microsomal fractions of first and second internodes of mechanically stimulated plants. This was true even though only the first internode was mechanically stimulated. Of the microsomal free fatty acids, mechanical perturbation induced an increase in myristic acid and large decreases in stearic, oleic, linoleic and linolenic acids. It also reduced the unsaturated:saturated ratio of the fatty acids. It induced a decrease in phosphatidyl choline but an increase in phosphatidyl ethanolamine. When the fatty acids were cleaved from the microsomal phospholipids, mechanical perturbation caused only a slight decrease in the unsaturated:saturated ratio and no significant changes in the double bond index. Mechanical perturbation induced an increase in the total microsomal protein and of membrane-associated latent IDPase. However, the activity of membrane-associated KCN-insensitive NADPH cytochrome-c reductase was decreased by mechanical perturbation. Treatment of the first internode with exogenous Ethrel produced results that were very similar in all respects to those obtained by mechanical perturbation. The factors inducing hardening against frost and drought, as achieved by mechanical perturbation and Ethrel treatment, are not only related to sterols or the polar head-groups of phospholipids but may also be related to the protein components, and all may have an effect on the fluidity of a bilayer membrane model. These data support the hypothesis that ethylene mediates thigmomorphogenesis and that mechanical perturbation of the first internode results in the acropetal transport of a translocatable thigmomorphogenetic factor.     

Thigmomorphogenesis: The Interaction of Wind and Temperature in the Field on the Growth of Phaseolus vulgaris L.

Field and laboratory experiments were performed to determinethe interaction of wind and temperature on thigmomorphogenesis(defined here as decreased elongation and increased internodalradial enlargement due to mechanical perturbation). Kidney beanplants (Phaseolus vulgaris L. cv. Red Cherokee Wax) were shelteredor exposed to wind for 10 days. Wind velocity, wind gusting,temperature, light and rainfall were measured for the durationof each of ten experiments. Stem elongation and diameter weremeasured and it was found that thigmomorphogenesis due to windoccurs naturally. There was more internodal secondary xylemproduced in the wind-exposed plants than in the sheltered plants.The experimental data were analyzed by multiple linear regressionand the wind was found to be a significant factor in the predictionof bean stem elongation and thickening. Laboratory experimentsshow that as wind velocity increases, thigmomorphogenesis increasesin an approximate linear fashion. Furthermore, it was foundthat low temperatures interact with mechanical perturbationto reduce the amount of thigmomorphogenesis, both in the fielddue to wind and in the laboratory due to rubbing. Other environmentalfactors do not seem to interact with the wind to modulate thigmomorphogenesiseven though these factors affect plant growth. Phaseolus vulgaris L., kidney bean, thigmomorphogenesis, wind, temperature, ethylene, multiple linear regression     

Thigmomorphogenesis: Ethylene evolution and its role in the changes observed in mechanically perturbed bean plants

Mechanical perturbations, in the form of either rubbing or wounding, cause ethylene evolution from bean internodes ( Phaseolus vulgaris L. cv. Cherokee Wax). This evolution begins 45 to 60 min after perturbation or wounding and peaks about 2 h later. Maximal thigmomorphogenesis occurs if internodes are perturbed when they are 10 mm or less in length. Maximal ethylene evolution, however, occurs in longer internodes. When one internode is perturbed, ethylene evolution is not observed from other internodes even though they respond thigmomorphogenetically by exhibiting decreased elongation. Ethylene evolution is apparently a result of increased 1-amino-cyclopropane-1-carboxylic acid (ACC) production after perturbation. Inhibitors of ACC and ethylene synthesis block increased radial growth but not reduced elongation. Ethylene may therefore be only one of several factors causing thigmomorphogenesis.     

Short-term responses of phloem transport to mechanical perturbation

Jaeger, C. H., Goeschl, J. D., Magnuson, C. E., Fares, Y. and Strain, B. R. 1988. Short-term responses of phloem transport to mechanical perturbation. - Physiol. Plant. 72: 588–594.
Phloem transport was monitored using a continuous stream of ¹¹CO₂-labelled air administered to one leaf while gamma detectors measured ¹¹C activity at intervals along the stem. The effect of gentle, non-injurious mechanical perturbation on phloem transport was tested in cotton ( Gossypium hirsutum L. cv. Stoneville 213). Mechanical stimuli such as shaking, localized vibration and gentle massage were applied while the plants were at isotope equilibrium. Localized phloem blockages were observed within 1–2 min of the stimuli. The blockages lasted from 6–55 min and full recovery of transport required 20–175 min. The effect of preconditioning to mechanical perturbation on phloem transport was tested in bush beans ( Phaseolus vulgaris L. cv. Cherokee Bush). Preconditioning of a bean seedling to gentle stem massage resulted in a shorter blockage response and quicker transport recovery period when the seedling was massaged during a ¹¹C tracer experiment compared to a control seedling. These results indicate that measurements of phloem transport on recently disturbed plants will probably show depressed phloem transport velocities. Measurements should be made after at least a 24-h disturbance-free recovery period.     

The effect of brassinosteroid and 2,4-D-L-amino acid conjugates on ethylene production by etiolated mung bean segments

Brassinosteroid (BR) an analogue of brassinolide was tested in combination with thirteen 2,4-dichlorophenoxyacetic acid-L-amino acid conjugates for its possible synergistic effects on ethylene production by etiolated mung bean (Vigna radiata L. Rwilcz cv. Berken) hypocotyl segments. When BR was used in combination with 2,4-D-L-amino acid conjugates the degree of enhanced ethylene production varied with the conjugate tested. In fact, the activity of the conjugate alone was directly related to its activity with BR.     

Is ethylene involved in regulation of root ferric reductase activity of dicotyledonous species under iron deficiency?

Most dicotyledonous species respond to Fe deficiency by developing some mechanisms known as Fe-deficiency responses. The role of ethylene in regulation of root ferric reductase activity of wild-type tomato (Lycopersicon esculentum L.) and its mutant Never ripe (Nr), bean (Phaseolus vulgaris L., cv. Bifeng 80-30), and cucumber (Cucumis sativus L., cv. Xintaimici) plants grown in nutrient solution without Fe supply was studied under controlled condition. The results show that: (i) the tomato mutant Nr, which is insensitive to ethylene, presented rapid increase in root ferric reductase activity after omitting Fe from the nutrient solution; (ii) the initial time for increase in root ferric reductase activity was earlier than that in ethylene production after onset of Fe deficiency in the three species; (iii) like cobalt (3 μM Co²⁺), an inhibitor for ethylene production, high concentration of zinc (50 μM Zn²⁺) and copper (5 μM Cu²⁺) also suppressed the increase in root ferric reductase activity of Fe-starved plants; (iv) under Fe-sufficient conditions, indol-3-butylric acid (IBA) stimulated root ferric reductase activity of cucumber and bean plants, and this stimulating effect could not be suppressed by aminoethoxyvinylglycine (AVG, an inhibitor for ethylene synthesis). These results suggested that ethylene might not be directly involved in the regulation of root ferric reductase activity of Fe-deficient dicotyledonous species.     

Responses of ethylene biosynthesis to saline stress in seedlings of eight plant species

The effect of salinity (100 mM NaCl) on ethylene metabolism in the early phase of vegetative development of several plant species has been investigated. The effects of saline treatment on shoot and root growth, ranged in sensitivity with respect to species: pepper (Capsicum annum L. cv Pairal) > tomato (Lycopersicon esculentum Mill. cv Malpica) > broccoli (Brassica oleraceae L. var. Italica Plenk. cv Marathon F1) ≅ lettuce (Lactuca sativa var. longifolia Lam. cv Inverna) ≅ melon (Cucumis melo L. cv Ruano F1, Roche type) > bean (Phaseolus vulgaris L. cv. Gator Green 15) ≅ spinach (Spinacia oleracea L. cv Boeing) > beetroot (Beta vulgaris L. var. crassa (Alef.) J. Helm. cv Detroit). After saline treatment, ethylene production increased 4.2-fold in pepper shoots. Significant increases were also found in shoots of tomato, broccoli and bean. In contrast, salinity decreased shoot ethylene production rate in melon, spinach, and beetroot. In roots, the general effect of salinity was a decrease in ethylene production, especially in broccoli and bean, except in tomato root, in which a sharp increase in ethylene production occurred. In general, saline treatment increased total ACC concentration in both shoot and root in most of the plant species examined, which was related to plant sensitivity to salinity. For example, pepper shoot was the most sensitive to saline treatment, showing the highest fresh weight inhibition and the highest increase in total ACC concentration (8.5-fold), while, beetroot was less affected by salinity and showed no effect on total ACC concentration in response to saline treatment.     

Thigmomorphogenesis: The Involvement of Auxin and Abscisic Acid in Growth Retardation Due to Mechanical Perturbation

When young bean plants are mechanically perturbed for up to10 days, they accumulate large amounts of an auxin-like substanceand increased amounts of ABA. Exogenous ethylene, applied inthe form of ethephon, produces the same result. Physiologicallymoderately high amounts of exogenously applied IAA or loweramounts of ABA cause the same sort of retardation of elongationthat is caused by either mechanical perturbation or exogenousethephon. Either mechanical perturbation or applied ethephonsignificantly retards the polar basipetal transport of ¹⁴C-IAAIt is proposed that mechanical perturbation of bean internodesinduces ethylene evolution which, in turn, induces the accumulationof high levels of IAA and the production of ABA, both of whichcontribute to the retardation of elongation of the internodes. (Received December 24, 1981; Accepted May 19, 1982)     

Abscission: role of abscisic Acid

The effect of abscisic acid on cotton (Gossypium hirsutum L. cv. Acala 4-42) and bean (Phaseolus vulgaris L. cv. Red Kidney) explants was 2-fold. It increased ethylene production from the explants, which was found to account for some of its ability to accelerate abscission. Absci is acid also increased the activity of cellulase. Increased synthesis of cellulase was not du to an increase in aging of the explants but rather was an effect of abscisic acid on the processes that lead to cellulase synthesis or activity.     

Interpreting Plant Responses to Clinostating: I. MECHANICAL STRESSES AND ETHYLENE

The severe epinasty and other symptoms developed by clinostated leafy plants could be responses to gravity compensation and/or the mechanical stresses of leaf flopping. Epinasty in cocklebur (Xanthium strumarium L.), tomato (Lycopersicon esculentum Mill.), and castor bean (Ricinus communis L.) is delayed by inhibitors of ethylene synthesis and action (aminoethoxyvinylglycine and Ag⁺), confirming the role of ethylene in clinostat epinasty. To test the possibility that clinostat mechanical stresses (leaf flopping) cause ethylene production and, thus, epinasty, vertical plants were stressed with constant, gentle, horizontal, or vertical shaking or with a quick, back-and-forth rotation (twisting). Clinostat leaf flopping was closely approximated but with a minimum of gravity compensation, by turning plants so their stems were horizontal, rotating them quickly about the stem axis, and then returning them to the vertical, repeating the treatment every four minutes (clinostat rotation time). None of these mechanical stresses produced significant epinasties, but vigorous hand-shaking (120 seconds per day) generated minor epinasties, as did Ag⁺ applied daily (concentrations high enough to cause leaf browning). Plants gently inverted every 20 minutes developed epinasty at about the same rate and to about the same extent as clinostated plants, but plants inverted every 20 minutes and immediately returned to the upright position did not become epinastic. It is concluded that clinostat epinasty is probably caused by disturbances in the gravity perception mechanism, rather than by leaf flopping.     

Identification of jasmonic acid and its methyl ester as gum-inducing factors in tulips

The purpose of this study was to identify endogenous factors that induce gummosis and to show their role in gummosis in tulip (Tulipa gesneriana L. cv. Apeldoorn) stems. Using procedures to detect endogenous factors that induce gum in the stem of tulips, jasmonic acid (JA) and methyl jasmonate (JA-Me) were successfully identified using gas–liquid chromatography–mass spectrometry. Total amounts of JA and JA-Me designated as jasmonates in tulip stems were also estimated at about 70–80 ng/g fresh weight, using deuterium-labeled jasmonates as internal standards. The application of JA and JA-Me as lanolin pastes substantially induced gums in tulip stems with ethylene production. The application of ethephon, an ethylene-generating compound, however, induced no gummosis although it slightly affected jasmonate content in tulip stems. These results strongly suggest that JA and JA-Me are endogenous factors that induce gummosis in tulip stems.     

春小麦水分胁迫响应中的ACC、MACC合成及乙烯的释放

水分胁迫使两个抗旱性不同的春小麦 (TriticumaestivumL .)品种“8139”(抗旱性较弱 )和“5 0 4”(抗旱性较强 )叶片ACC和MACC含量于胁迫初期下降后期升高 ,ACC合酶活性持续升高 ,乙烯释放量在 8139中下降而在5 0 4中先大幅升高而后下降。两种作用效果相反的抑制剂MGBG (抑制SAMDC活性 )和AOA (抑制ACC合酶活性 )均明显影响了两品种春小麦叶片以上各指标的变化。结果表明 ,水分胁迫下作物乙烯的释放量并不与其合成直接前体ACC的量成正相关 ;胁迫乙烯在抗性品种中于胁迫初期的升高可能是植物胁迫信号传导的响应之一 ,是一种干旱适应现象 ,可能与作物的干旱忍耐形成有关 ,而MACC具有调节胁迫乙烯释放的特殊生理作用。     

Contribution of corticular photosynthesis to bud development in African baobab (Adansonia digitata L.) and Castor bean (Ricinus communis L.) seedlings

The African baobab (Adansonia digitata L.) and castor bean (Ricinus communis L.) are drought resistant green-stemmed succulent plants which grow in the arid and semi-arid regions of Africa. Photosynthesis in the stems of green-stemmed plants is known to contribute to plant carbon gain especially during leafless periods. To study the contribution of stem photosynthesis in stem succulent plants, the height and stem diameter of baobab and castor bean plants grown in the greenhouse were measured. The plants were completely defoliated and subjected to different treatments: Watered with open stems (WO), watered and stems covered with aluminium foil (WC) to achieve 100% light exclusion, drought and open (DO) and drought and covered (DC). Stem coverage with aluminium foil resulted in a higher stem height and diameter during drought for baobab with similar trends seen in castor bean. Light exclusion resulted in a significantly lower bud DW production and enrichment in ¹³C in bud dry matter of castor bean and in stem dry matter of baobab. These show that corticular photosynthesis contributes in carbon gain in these species.     

乙烯对UV-B辐射诱导蚕豆气孔关闭的调控效应(英文)

UV-B辐射作为一种重要的环境信号影响着植物的生长与发育,它能够调控气孔运动和诱导乙烯产生.该试验利用乙烯生物合成抑制剂和乙烯受体抑制剂处理蚕豆叶片表皮条,结合气孔开度分析和乙烯释放量测定,研究乙烯在UV-B辐射调控表皮条气孔运动中的作用.结果发现,将蚕豆叶片表皮条置于0.8 W·m-2的UV-B辐射下1～4 h,乙烯生成和气孔关闭均被显著诱导,且乙烯释放峰先于气孔关闭的起始;乙烯生物合成抑制剂和乙烯受体抑制剂处理均能显著逆转UV-B辐射诱导的气孔关闭;外源乙烯处理也能模拟UV-B辐射的效应诱导可见光下蚕豆表皮条的气孔关闭.可见,乙烯介导了UV-B辐射诱导的蚕豆气孔关闭.     

Effect of regurgitant from Leptinotarsa decemlineata on wound responses in Solanum tuberosum and Phaseolus vulgaris

The effect of regurgitant from Leptinotarsa decemlineata Say larvae on wound-induced responses was studied using two plant species, Solanum tuberosum L. and Phaseolus vulgaris L. Wounding of one leaf of intact S. tuberosum plants differentially affected ethylene production and activities of peroxidase and polyphenol oxidase. Only polyphenol oxidase activity was stimulated by wounding in both wounded and systemic leaves. Peroxidase activity was not affected by wounding. Wounding caused only a transient increase of ethylene production from wounded leaves. The application of regurgitant to wound surfaces stimulated ethylene production as well as activities of peroxidase and polyphenol oxidase in both wounded and systemic leaves. Wounding significantly enhanced ethylene production and polyphenol oxidase activity in wounded and systemic leaves of P. vulgaris . The application of regurgitant caused an amplification of ethylene production, peroxidase activity, and polyphenol oxidase activity, in both wounded and systemic leaves of bean plants. Several substances were tested for their role as possible endogenous signals in P. vulgaris . Hydrogen peroxide and methyl jasmonate appeared as potential local and systemic signals of ethylene formation in wounded bean plants. Local ethylene production in leaf discs was differentially affected by the regurgitant application in potato versus bean plants. While all tested concentrations of regurgitant caused stimulation of ethylene formation from potato leaf discs, ethylene production was completely inhibited by increasing concentrations of the regurgitant in bean leaf discs. Our data present evidence that ethylene may play an important role in the interaction between plants and herbivores at the level of recognition of a particular herbivore leading to specific induction of signalling cascades.     

Absorption and translocation of Cd in bush beans (Phaseolus vulgaris)

A series of experiments was conducted to examine some factors affecting the absorption and translocation of Cd in young bean plants ( Phaseolus vulgaris L. cv. Bulgarian). Absorption of Cd by roots was reduced in the presence of other cations of increasing valency or ionic radii. Reduced absorption was also found in the presence of EDTA. Concentration of Cd in exudates from excised stems increased with increased passage of Cd solutions and approached the concentration in the external medium (4.5 μ M Cd). This was apparently associated with saturation of adsorption sites in the stems. The stem behaved as a cation exchange column resulting in a chromatographic distribution of Cd towards the top of the plant. These experiments indicate that Cd existed in the xylem fluid as a free or weakly complexed cation. Additional experiments showed that the total amount of Cd absorbed by bean plants was elevated by inducing higher transpiration rates. The effect of water flux on Cd transport indicated apoplastic flow to the stele.     

Inhibition of IAA-induced ethylene production in etiolated mung bean hypocotyl segments by 2,3,5-triiodobenzoic acid and 2-(p-chlorophenoxy)-2-methyl propionic acid

The effect of two auxin antagonists, 2,3,5-triiodobenzoic acid (TIBA) and 2-( p -chlorophenoxy)-2-methyl propionic acid (CMPA) on IAA-induced ethylene production in etiolated mung bean hypocotyl ( Vigna radiata L. Rwilcz cv. Berken) segments was studied. Both TIBA and CMPA inhibited IAA-induced ethylene production and CO₂ production at concentrations from 0.001 m M to 0.1 m M and 0.01 m M to 1.0 m M , respectively. The optimum concentration for inhibition of ethylene production by TIBA was 0.05 m M and CMPA was 0.5 m M . At the optimum concentration of TIBA and CMPA, there was a significant decrease in IAA-induced ethylene production without a decrease in respiration rates below control levels. After 18 h, mung bean hypocotyl segments treated with 0.05 m M TIBA for 6 h or 0.5 m M CMPA for 8 h showed a maximum inhibition of IAA-induced ethylene production. Treatments longer than 8 h caused no further inhibition. The uptake of [¹⁴C]-naphthaleneacetic acid by mung bean segments was greatly reduced by the addition of either TIBA (0.05m M ) or CMPA (0.5 m M ) to the incubation media. The results of treatment sequences showed that TIBA needed to be applied prior to IAA in order to inhibit IAA-induced ethylene production, but CMPA caused the same inhibitory effect whether applied before or after IAA treatment. These findings provide evidence that TIBA inhibits auxin-induced ethylene production in etiolated mung bean hypocotyl segments by blocking auxin movement into the tissue whereas CMPA may work on both auxin transport and action.     

Ethylene Biosynthesis and Cadmium Toxicity in Leaf Tissue of Beans (Phaseolus vulgaris L.)

Stress ethylene production in bean (Phaseolus vulgaris L., cv. Taylor's Horticultural) leaf tissue was stimulated by Cd²⁺ at concentrations above 1 micromolar. Cd²⁺-induced ethylene biosynthesis was dependent upon synthesis of 1-aminocyclopropane-1-carboxylic acid (ACC) by ACC synthase. Activity of ACC synthase and ethylene production rate peaked at 8 h of treatment. The subsequent decline in enzyme activity was most likely due to inactivation of the enzyme by Cd²⁺, which inhibited ACC synthase activity in vitro at concentrations as low as 0.1 micromolar. Decrease in ethylene production rate was accompanied by leakage of solutes and increasing inhibition of ACC-dependent ethylene production. Ca²⁺, present during a 2-hour preincubation, reduced the effect of Cd²⁺ on leakage and ACC conversion. This suggests that Cd²⁺ exerts its toxicity through membrane damage and inactivation of enzymes. The possibility of an indirect stimulation of ethylene biosynthesis through a wound signal from injured cells is discussed.     

An auxin-responsive 1-aminocyclopropane-1-carboxylate synthase is responsible for differential ethylene production in gravistimulated<Emphasis Type="Italic"> Antirrhinum majus</Emphasis> L. flower stems

The regulation of gravistimulation-induced ethylene production and its role in gravitropic bending was studied in Antirrhinum majus L. cut flower stems. Gravistimulation increased ethylene production in both lower and upper halves of the stems with much higher levels observed in the lower half. Expression patterns of three different 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) genes, an ACC oxidase (ACO) and an ethylene receptor (ETR/ERS homolog) gene were studied in the bending zone of gravistimulated stems and in excised stem sections following treatment with different chemicals. One of the ACS genes (Am-ACS3) was abundantly expressed in the bending zone cortex at the lower side of the stems within 2 h of gravistimulation. Am-ACS3 was not expressed in vertical stems or in other parts of (gravistimulated) stems, leaves or flowers. Am-ACS3 was strongly induced by indole-3-acetic acid (IAA) but not responsive to ethylene. The Am-ACS3 expression pattern strongly suggests that Am-ACS3 is responsible for the observed differential ethylene production in gravistimulated stems; its responsiveness to IAA suggests that Am-ACS3 expression reflects changes in auxin signalling. Am-ACS1 also showed increased expression in gravistimulated and IAA-treated stems although to a much lesser extent than Am-ACS3. In contrast to Am-ACS3, Am-ACS1 was also expressed in non-bending regions of vertical and gravistimulated stems and in leaves, and Am-ACS1 expression was not confined to the lower side cortex but evenly distributed over the diameter of the stem. Am-ACO and Am-ETR/ERS expression was increased in both the lower and upper halves of gravistimulated stems. Expression of both Am-ACO and Am-ETR/ERS was responsive to ethylene, suggesting regulation by IAA-dependent differential ethylene production. Am-ACO expression and in vivo ACO activity, in addition, were induced by IAA, independent of the IAA-induced ethylene. IAA-induced growth of vertical stem sections and bending of gravistimulated flowering stems were little affected by ethylene or 1-methylcyclopropene treatments, indicating that the differential ethylene production plays no pivotal role in the kinetics of gravitropic bending.