Cloning and Sequencing of Phytoplasma Ribosomal DNA (rONA) Associated with Kerala Wilt Disease of Coconut Palms

Using the polymerase chain reaction (PCR) the 165 rRNA gene of phytoplasma associated with Kerala wilt disease of coconut palm (Cocos nucifera L) was amplified from infected leaf samples. Within the three universal primer pairs P1/P6, P1/P7and P41 P7, the primer pair P4/P7 only showed an amplification of 650 bp DNA fragment. 5ince P4/P7 amplifies the 16S-23S intergenic spacer region of 165 rRNA gene, the PCR product 650 bp of Kerala wilt disease palm indicates the phytoplasma DNA. The amplified fragment was sequenced and deposited in Genbank data library (Accession No. AY158660). The absence of restriction sites for Bcll and Rsa/l in 650 bp indicates phytoplasmic nature of DNA and its strain difference. A comparison of the 650 bp sequence with other phytoplasmas and its restriction profile indicates Kerala wilt disease phytoplasma as a separate 165 rRNA group in the classification of phytoplasmas. To our knowledge, this report records the first finding of the phytoplasma DNA using universal primers and its sequence analysis in coconut palms of Kerala, south India.     

Detection and Molecular Characterization of Phytoplasma associated with Lethal Yellowing Disease of Coconut Palms in Cuba

Lethal Yellowing (LY) disease of coconut palm (Cocos nucifera L.) in Cuba has been reported since the end of the 19th century. In order to ascertain the presence of phytoplasmas associated with this disease, leaf samples were taken from plants showing typical disease symptoms and assayed for the LY agent by the polymerase chain reaction (PCR) using LY‐specific primers. Selected PCR amplification products were cloned, sequenced and compared to that of a Mexican LY isolate from the Yucatán region. The results obtained confirm the presence of LY phytoplasma in Cuba. Cuban and Mexican isolates show an overall high degree of sequence similarity with occasional point mutations and small deletions or insertions. Based on these identified genetic differences, LY isolates from the Havana and the Yucatán region cluster together and apart from isolates originating at Maisí in eastern Cuba.     

Light-Induced Stomatal Opening Is Affected by the Guard Cell Protein Kinase APK1b

Guard cells allow land plants to survive under restricted or fluctuating water availability. They control the exchange of gases between the external environment and the interior of the plant by regulating the aperture of stomatal pores in response to environmental stimuli such as light intensity, and are important regulators of plant productivity. Their turgor driven movements are under the control of a signalling network that is not yet fully characterised. A reporter gene fusion confirmed that the Arabidopsis APK1b protein kinase gene is predominantly expressed in guard cells. Infrared gas analysis and stomatal aperture measurements indicated that plants lacking APK1b are impaired in their ability to open their stomata on exposure to light, but retain the ability to adjust their stomatal apertures in response to darkness, abscisic acid or lack of carbon dioxide. Stomatal opening was not specifically impaired in response to either red or blue light as both of these stimuli caused some increase in stomatal conductance. Consistent with the reduction in maximum stomatal conductance, the relative water content of plants lacking APK1b was significantly increased under both well-watered and drought conditions. We conclude that APK1b is required for full stomatal opening in the light but is not required for stomatal closure.     

Role of Pressure in Xylem Transport of Coconut and Other Palms

The significance of root pressure in the transport of xylem sap has been investigated in Cocos nucifera L. and a few other palms. Despite the fact that excised palm roots can generate considerable pressures in situ, the quantity of water transported is only a small fraction of the demand resulting from transpiration. Most water transport is induced by negative pressure gradients, as in other higher plants. The development of considerable negative pressures has been demonstrated both directly and indirectly. Acoustic detection was used for the first time to monitor cavitation in water-stressed Cocos leaves. Its detection implies the ready disruption of xylem sap under these tensions. We suggest that root pressure might serve to refill cavitated xylem conduits when water is abundantly available and transpiration practically zero. However, little or no positive pressure could be demonstrated in intact palms subjected to low water stress: experimentally.     

Suppression of Stomatal Opening in Leaves Treated with Abscisic Acid

Small doses of abscisic acid (approximately 0.02 µg cm^-2of leaf) applied to the leaf surface as a 10^-4 M solution causedmarked stomatal closure in Xanthium pennsylvanicum, and theeffect persisted for up to 9 days after application. Similareffects were found when 10^-4 M abscisic acid was supplied todetached tobacco leaves via their petioles. CO²-free air didnot cause a reversal of the closure, and it was therefore concludedthat the effect was not due simply to an increase in the intercellularCO²concentration; a more direct effect on the stomatal apparatusis suggested. It is considered that abscisic acid could playan endogenous role in the control of stomatal aperture, andthat this, and/or related substances, might be more useful as‘anti-transpirants’ than the phytotoxic substancescurrently employed for this purpose.     

Zinc Requirement for Stomatal Opening in Cauliflower

Zn deficiency induced increases in epicuticular wax deposits, lamina thickness, degree of succulence, water saturation deficit, diffusive resistance, and proline accumulation and decreases in carbonic anhydrase activity, water potential, stomatal aperture, and transpiration in the leaves of cauliflower (Brassica oleracea L. var botrytis cv Pusa) plants. Restoration of Zn supply to the deficient plants increased stomatal aperture, transpiration, and carbonic anhydrase activity significantly within 2 h. However, leaf water potential in the Zn-deficient plants did not recover within 24 h after resupply of Zn. The guard cells in epidermal peels from the Zn-deficient leaves had less K+ than those from the controls. Stomatal aperture in the epidermal peels from Zn-deficient leaves was 64% less than in the controls when the epidermal strips were floated on 125 mM KCl. Supplementing the ambient medium 25 mM KCl with ZnCl2 enhanced stomatal aperture in both control and Zn-deficient peels, and the effect was significant in the latter. The observations indicate involvement of Zn in stomatal opening, possibly as a constituent of carbonic anhydrase needed for maintaining adequate [HCO3-] in the guard cells, and also as a factor affecting K+ uptake by the guard cells.     

Studies of the Mechanism by which Daminozide (B9) Inhibits Stomatal Opening

The mechanism by which the synthetic growth retardant daminozide(B9) inhibits stomatal opening has been investigated, and itsaction compared with that of the naturally occurring growthhormone, abscisic acid. The inhibition of stomatal opening bylow concentrations of daminozide in normal air is largely reversedin CO₂-froe air, suggesting that the main effect is due to anincrease in internal CO₂ concentratiom This is supported byobservations of a raised level of CO₂ compensation in treatedleaves. The CO₂-independent light reaction of stomata does,however,also appear to be inhibited by dammozide. Higher concentrationscause damage to cells of the epidermis, and such effects couldalso be found in normal practical use of the growth retardant. The mechanism of action of daminozide on stomata has littlein common with that of abscisic acid, which does not operateby increasing the internal CO₂ concentration and does not inducecellular damage. Daminozide cannot be recommended for practicaluse as an antitrans-pirant because of its mechanism of action,which is likely to cause a decrease in the efficiency of wateruse.     

Nuclear DNA Amounts in Palms (Arecaceae)

Nuclear DNA amounts are reported for 83 species and 53 genera of palms, covering all of the six subfamilies. 4C DNA contents range between 3.89 and 55.62 pg in diploids, showing an approximate 14.3-fold variation in genome size. Polyploids have DNA contents of up to 156.40 pg/4c which demonstrates a 40.2-fold variation. Diploids with high DNA contents occur in three subfamilies of palms (Coryphoideae, Calamoideae, Arecoideae), and seem to be further restricted to particular tribes or subtribes (Thrinacinae, Borasseae, Lepidocaryeae, Caryoteae, some subtribes of Areceae). Palms from the subfamilies Nypoideae and Phytelephantoideae have the lowest DNA amounts, followed by the Phoeniceae and the Corypheae: Livistoninae from the subfamily Coryphoideae. Although DNA amounts in some genera and subtribes are usually constant, e.g., in Phoenix, Phytelephas, the Livistoninae, Dypsidinae, diploid Butiinae), considerable variation occurs at the diploid level in some large and apparently actively evolving genera such as Chamaedorea, Pinanga, Cenoma and possibly Bactris. Formaldehyde fixation is recommended for palms, as conventional ethanol-acetic acid fixation has proved to be unsuitable for DNA estimation of Feulgen-stained nuclei by microdensitometry, since it can lead to errors up to 2.5-fold in extent. Chromosome counts are reported for 72 of the species studied, of which 42 are new.     

细胞骨架参与乙酰胆碱诱导的气孔开放

动物系统中,乙酰胆碱（Ａｃｈ）是胆碱能系统中的主要成分,近上的研究表明高等植物体内均有Ａｃｈ、胆碱乙酰基转移酶和乙酰胆碱酯酶,以及乙酰胆碱受本的存在,这些物质调节着生命吕的许多重要过程,如气孔运动等。以前的研究表明微管、微丝参与了气孔的运动。本研究了Ａｃｈ调控的气孔开放是否与微管、微丝的结构相关联。结果表明Ａｃｈ可以在ＫＣｌ溶液和无ＫＣｌ溶液中诱导气孔开放;同时Ａｃｈ还能在无ＫＣｌ溶液中促进保卫     

Control of Root Rot and Wilt caused by Pythium myriotylum in Kangaroo Paw (Anigozanthos)

Control of root rot and wilt on Kangaroo Paw (Anigozanthos) caused by Pythium myriotylum was tested, using artificial inoculations, in greenhouse experiments. Disease incidence in the inoculated but non‐treated plots was 74% in Expt A (1999) and 46% in Expt B (2000). Disease incidence was significantly reduced by treatments with fenamidon (26 and 70%) and Terraclor SuperX (65 and 41%) in Expts A and B, respectively, in comparison with the non‐treated plots. Disease incidence was reduced with Ridomil Gold by 7 and 48% in Expts A and B, respectively; with Dynone by 22 and 41% in Expts A and B, respectively; with Tachigaren by 20% in both experiments; and with Trichoderma harzianum treatment, disease incidence was reduced by 9% only in Expt B. Yields (recorded only in Expt B) obtained in plots inoculated with P. myriotylum were significantly lower than in the non‐inoculated control plots. The most efficient treatment was fenamidon, with 324% more flowers than in the non‐treated control. The number of flowers in terraclor SuperX, Ridomil Gold, Dynone and T. harzianum treatments was higher by 285, 249, 234 and 235% than in the non‐treated control. In vitro tests demonstrated that the most effective doses, causing 50% inhibition of mycelial growth (ED₅₀), were 0.09, 0.4, 0.4 and 4 μg/ml for the fungicides fenamidon, Ridomil Gold (mefenoxam), Terraclor SuperX (etridiazole + quintozene) and Dynone (prothiocarb), respectively.     

Effects of Organic Nutrients and Hormones on Growth and Development of Tissue Explants from Coconut (Cocos nucifera) and Date (Phoenix dactylifera) Palms Cultured in vitro

The growth response (increase in weight) of cultured explants from seedling date (Phoenix dactylifera L.) and mature coconut (Cocos nucifera L. cv. Malayan Dwarf) palms to source and concentration of organic nitrogen. carbohydrate, auxins, cytokinins and gibberellins was examined. Growth was strongly stimulated by the presence of auxins (10^?7 to 10^?6M), cytokinins (10^?6 to 10^?5M), high concentrations of sucrose (0.2 M), and in the absence of NH₄Cl, by organic sources of reduced nitrogen. Higher concentrations of auxin (2,4-D or NAA at 10^?6 to 10^?5M) which still stimulated growth of Phoenix tissue, proved inhibitory to growth of freshly excised Cocos tissues. Explants from both palms initiated roots when subcultured on a medium with increased levels of auxin (NAA, 2.5 × 10^?6 to 2.5 × 10^?5M) and reduced levels of cytokinin (6-BAP, 5 × 10^?8M). Isolated roots excised from these explants continued growth and produced new laterals when subcultured on media with GA₃ (5 × 10^?7M) and reduced levels of auxin, cytokinin, and either minerals or sucrose.     

Effect of Photoperiod on Stomatal Opening in Vicia faba

Stomatal apertures in darkness and subsequent average opening rates in light were measured in Vicia faba leaf discs throughout the nyctoperiods for plants grown on three light:dark cycles (8:16, 12:12, and 16:8). The time course of opening in darkness depended on the specific light:dark cycle with the maximum aperture always occurring at the time the lights normally went on. The light-induced opening rate was also maximum at the end of the nyctoperiod.     

Acid-Induced Stomatal Opening in Commelina communis and Vicia faba

The effects of H^$ and fusicoccin (FC) on stomatal opening inthe dark were investigated using epidermal strips of Commelinacommunis and Vicia faba cv. Ryosai Issun. Citrate-phosphatebuffer induced maximal opening of stomata at pH 3.0 when testedover the range of 2.7 to 5.0. HCl at 1 mM also induced stomatalopening without appreciable accumulation of K^$ in the guardcells. After 4 hr treatment with 10 µM FC, stomata openedwith concomitant accumulation of K^$ in the guard cells, although1–2 hr treatment caused opening without concomitant K^$increase. These results suggest that stomatal opening can be caused bysalt accumulation and/or changes of the physicochemical conditionsin the cell wall of the guard cells due to high acidity. ¹ Present address: Biological Laboratory, Faculty of Education,Nagasaki University, Nagasski 852, Japan. (Received April 30, 1982; Accepted July 17, 1982)     

Stilt Root Structure in the Neotropical Palms Iriartea deltoidea and Socratea exorrhiza1

Many arborescent palms develop a stilt root cone that provides increased mechanical support on steep hillsides, better root aeration under waterlogged conditions, resprouting after mechanical damage, and rapid stem elongation. However, for most species the role of stilt roots is not well understood. We determined how palm size and slope inclination affected the structure of stilt roots in the neotropical palms Iriartea deltoidea and Socratea exorrhiza. We expected palm height to be lower on steep slopes because the effectiveness of root support could decrease as slope inclination increases, and thus, the structure of the root cone would vary mostly with slope. Alternatively, if stilt root development is determined by palm size, their production should match stem height, with taller palms having larger cones. We measured the stilt root cone of 31 Iriartea and 36 Socratea palms in San Ramón Biological Reserve and Golfito Wildlife Refuge, Costa Rica. We divided the cone into five variables (horizontal projections of the cone base and stilt root height up and down the slope, and width of the cone base), from which we extracted the first two principal components and used them to measure the effects of slope and palm size on stilt root structure. We found stilt roots to be determined by palm size, not by slope conditions. Stilt roots matched palm size, with larger palms having taller and larger cones, maintaining stability under diverse slope conditions.     

Latent Periods of Cytokinin-induced Stomatal Opening in the Sunflower Leaf

Spraying a zeatin solution on the mature leaf of an intact sunflowerplant (Helianthus annuus L. cv. Large Russian) caused a two-folddecrease in leaf temperature during the 20 min after treatment.Scanning electron microscope photographs taken 2 min after cytokinintreatment, however, showed a small but distinct stimulationof the opening of the central pore of the stomata, indicatingthat the latent period of cytokinin-induced stomatal openingis only 2 min. Five minutes after treatment with zeatin, thestomata at both the central pore and the outer ledge had openedwidely. On the other hand, zeatin riboside treatment did notshow a distinct effect on the stomatal opening within theseperiods. The stomata of a water-sprayed leaf closed within 1min after the treatment. (Received January 7, 1981; Accepted June 12, 1981)     

Effect of the Mesophyll on Stomatal Opening in Commelina communis

The effect of a number of factors on the opening of stomatain the intact leaf and in the isolated leaf epidermis of Commelinacommunishas been investigated. Stomata in the intact leaf opened widein the light and closed rapidly on transfer to the dark. Theywere also sensitive to CO₂. In contrast, stomata in isolatedepidermis floated on an incubation solution containing 100 molm^–3KCl responded neither to light nor CO₂. They opened as widelyas those in the intact leaf when treated with fusicoccin. Stomata in isolated epidermis opened almost as wide as thosein the intact leaf when they were incubated with isolatedmesophyllcells in the light. The solution in which the mesophyll cellswere incubated was separated by centrifugation. Themedium fromcells previously incubated in the light caused the stomata inisolated epidermis to open but that from cells kept inthe darkhad no effect. A similar effect was observed when isolated chloroplastswere incubated with the isolated epidermis.However, the supernatantfrom the chloroplast suspension had no significant effect onstomatal opening. These results indicate that the mesophyll plays an importantrole in stomatal opening in the light. The mesophyll appearstoproduce in the light, but not in the dark, a soluble compoundwhich moves to the guard cells to bring about stomatal opening.Theexperiments with isolated chloroplasts suggest that this substanceis a product of photosynthesis. Key words: Commelina communis, stomata, light, mesophyll     

Periodic Nocturnal Stomatal Opening of Citrus in a Steady Environment

In a steady environment, leaf stomates of sour orange (Citrus aurantium L.) remained closed in the dark, whereas those of rough lemon (Citrus jambhiri Lush) underwent several 20-minute periods of pronounced opening at intervals of 60–85 minutes. Peak nocturnal opening occurred about 10 minutes after the cycle began, as shown by a change in leaf diffusion resistance from 60 s cm^?1 (closed) to 2 s cm^minus;1 (open). This minimal leaf resistance equals that obtainable in rough lemon in strong illumination (26 kilolux). The transitory stomatal opening detected by leaf resistance measurements was corroborated by measurements of leaf temperature and leaf thickness. During nocturnal opening, leaf temperature was 2°C below the value representing non-cycling periods. Also, minimal leaf thickness coincided with maximal stomatal opening. The triggering mechanism for nocturnal stomatal opening appears to originate within the plant, since the environmental factors of air temperature, humidity, carbon dioxide concentration, and substrate (aerated water culture) were held steady.