Molecular biology of wound-inducible proteinase inhibitors in plants

Abstract. The techniques of molecular biology are being employed to investigate at the gene level the systemically mediated, wound-induced accumulation of two defensive proteinase inhibitor proteins in plant leaves. These techniques have added a new dimension to biochemical and physiological studies already underway to understand the mechanism of induction by wounding. The acquisition of cDNAs from the RNAs coding for the two inhibitors facilitated studies of mRNA synthesis in leaves in response to wounding, and provided probes to obtain wound-inducible proteinase inhibitor genes from tomato ( Lycopersicon esculentum ) and potato (Solarium tuberosum) genomes. Successful transformations of tobacco plants with fused genes, containing the 5' and 3' regions of the inhibitor genes with the open reading frame of the chloramphenicol acelyltransferase ( cat ) gene, have provided a wound-inducible chloramphenicol acetyltransferase (CATase) activity with which to seek cis- and transacting elements that regulate wound-inducibility to help to understand the interaction of cytoplasmic and nuclear components of the intracellular communication systems that activate the proteinase inhibitor genes in response to wounding by insect pests.     

Vocal morphology of the Physalaemus pustulosus species group (Leptodactylidae): morphological response to sexual selection for complex calls

Most male frogs in the genus Physalaemus produce a whine-like advertisement call. Male P. pustulosus , however, add chucks to the call. This enhances the attractiveness of the call to females, and has evolved under the influence of sexual selection despite the increased predation risk from the frog-eating bat ( Trachops cirrhosus ). This complex call is unusual, if not unique, among anurans because the two call components overlap in time. Here we investigate the morphological changes responsible for the production of complex calls.
The Physalaemus purtulosus species group consists of four species. Physalaemus pustulosus and P. petersi are sister species, and recently it has been shown that P. petersi produces chucks. Physalaemus coloradorum and P. purtulatus are sister species and neither is known to produce chucks. Two laryngeal characters vary within the species group. Physalaemus pustulosus has a large fibrous mass (FMI), whose vibration is responsible for production of the chuck. This mass is much smaller in the other three species. In P. pustulosus and P. petersi the FMI is anchored dorsally, deep within the bronchial process, the attachment is more extensive in P. pustulosus. Neither P. pustulatus nor P. coloradorum have such a dorsal attachment associated with their FMI. This character is responsible for allowing the FMI to vibrate independently of the vocal cords, that is, for the production of the complex call. Thus the morphological changes responsible for the evolution of this unusual behavioural innovation, the complex call, are gradual, and almost trival, in nature. This study also shows that the primitive condition of the larynx of the P. pustulosus and P. petersi ancestor, was predisposed to the production of complex calls.
We also document ontogenetic and sexually dimorphic patterns in larynx structure.     

Malate Accumulates in the Vacuole of Chara australis During Uptake of Ammonium From Chloride-free Solution

Internodal cells of Chara australis can accumulate ammoniumto high concentrations (10 to 70 mol m^–3) in their vacuoles.When Cl^– is included in the bathing solution, changesin the cellular concentrations of ammonium, K⁺, Cl^– andNa⁺ have been shown to meet the requirements for electroneutralityand to account for the changes in vacuolar osmotic pressureassociated with ammonium uptake. If accumulation occurs in theabsence of external Cl^–, however; changes in the inorganicions do not meet these criteria. Malate is found in the vacuolesof cells accumulating amine in the absence of external Cl^–and its presence (at 0·5 to 8·5 mol m^–3)allows us to account for electroneutrality and for changes inthe osmotic potential. Key words: Malate, Chara, electroneutrality, ammonium     

Aluminium Toxicity in Roots: An Investigation of Spatial Sensitivity and the Role of the Root Cap

The primary symptom of aluminium (Al) toxicity in higher plantsis inhibition of root growth. In this study, we investigatedthe spatial sensitivity of maize (Zea mays L.) roots to Al.A divided-chamber technique indicated that only exposure ofthe terminal 10 to 15 mm of the root to Al resulted in inhibitionof growth. Application of Al to all but this apical region ofthe root had little or no effect on growth for 24 h and causedminimal damage to the root tissue. Small agar blocks infusedwith Al were then applied to discrete areas of the apex of maizeroots to determine which section (root cap, meristem or elongationzone) was more important to Al-induced inhibition of growth.The terminal 20 to 30 mm of root (root cap and meristem) mustbe exposed to Al for inhibition. Application of Al to the 30mm of root proximal to this terminal zone (elongation zone)resulted in damage to the root tissue but no significant inhibitionof growth. Therefore, the visible injuries incurred by rootsduring Al-stress are not associated directly with the inhibitionof root growth. Furthermore, removal of the root cap had noeffect on the Al-induced inhibition of root growth in solutionexperiments and argues against the root cap providing protectionfrom Al stress or serving an essential role in the mechanismof toxicity. We suggest that the meristem is the primary siteof Al-toxicity. Key words: Aluminium, toxicity, root growth, root cap     

Rapid calcium exchange for protons and potassium in cell walls of Chara

Net fluxes of Ca²⁺, H⁺ and K⁺ were measured from intact Chara australis cells and from isolated cell walls, using ion-selective microelectrodes. In both systems, a stimulation in Ca²⁺ efflux (up to 100 nmol m^?2 s^?1, from an influx of ～40 nmol m^?2 s^?1) was detected as the H⁺ or K⁺ concentration was progressively increased in the bathing solution (pH 7.0 to 4.6 or K⁺ 0.2 to 10mol m^?3, respectively). A Ca²⁺ influx of similar size occurred following the reverse changes. These fluxes decayed exponentially with a time constant of about 10 min. The threshold pH for Ca²⁺ efflux (pH 5.2) is similar to a reported pH threshold for acid-induced wall extensibility in a closely related characean species. Application of NH₄⁺ to intact cells caused prolonged H⁺ efflux and also transient Ca²⁺ efflux. We attribute all these net Ca²⁺ fluxes to exchange in the wall with H⁺ or K⁺. A theoretical treatment of the cell wall ion exchanges, using the ‘weak acid Donnan Manning’ (WADM) model, is given and it agrees well with the data. The role of Ca²⁺ in the cell wall and the effect of Ca²⁺ exchanges on the measured fluxes of other ions, including bathing medium acidification by H⁺ efflux, are discussed.     

Craniofacial ontogeny in centrosaurine dinosaurs (Ornithischia: Geratopsidae): taxonomic and behavioral implications

Centrosaurine ceratopsians are characterized by well developed nasal horncores or bosses, relatively abbreviated supraorbital horncores or bosses, and adorned parietosquamosal frills. Recent study of several paucispecific (low diversity) bonebed assemblages in Alberta and Montana has contributed greatly to our understanding of ontogenetic and taxonomic variation in the skulls of centrosaurines. Relative age determination of centrosaurines is now possible through examination of ontogenetic change in several characters, including the surface bone morphology of specific skeletal elements. The within-group taxonomy of centrosaurines is based almost entirely on characters of the skull roof, relating particularly to horns and frills. Juvenile and sub-adult centrosaurines are characterized by relatively simple, unadorned skulls compared to their adult counterparts. As in numerous living taxa, the cranial ornaments of centrosaurines developed late in ontogeny, as individuals approached or attained adult size. An important implication arising directly from this study is that juvenile and sub-adult centrosaurines are difficult to distinguish taxonomically at the specific level. Two monospecific genera represented only by immature materials, Brachyceratops montanensis and Monoclonius crassus , cannot be defended and should be considered nomina dubia . The late ontogenetic development and diverse taxonomic variation of horn and frill morphologies support the contention diat these structures are best interpreted as reproductive characters employed in mate competition.     

Pre-breeding diet influences ornament size in the Rhinoceros Auklet Cerorhinca monocerata

Understanding the mechanisms that influence variation in sexually selected ornaments in seabirds has been challenging owing to the difficulty of capturing and sampling individuals outside of the breeding period when ornaments are usually grown. Stable carbon (δ¹³C) and nitrogen (δ¹⁵N) isotopes were used to examine the influence of pre-breeding diet composition on ornament size in the Rhinoceros Auklet Cerorhinca monocerata , a socially monogamous seabird that breeds in the North Pacific. We analysed stable isotopes in adult feathers grown during the pre-alternate moult, which allowed us to infer diet composition during the pre-breeding (February–March) period. Females that fed more on inshore fish had larger horns than females that fed more on euphausiids (also known as krill; Euphausiacea). Body size was a stronger predictor of horn height in males than females, suggesting that ornaments may serve as different signals for each sex. This study provides evidence that diet during the pre-breeding period can influence ornament size and emphasizes the importance of understanding individual ecology throughout the annual cycle for determining the factors that influence mate choice and fitness.