Plant phenotypic plasticity in a changing climate.

Climate change is altering the availability of resources and the conditions that are crucial to plant performance. One way plants will respond to these changes is through environmentally induced shifts in phenotype (phenotypic plasticity). Understanding plastic responses is crucial for predicting and managing the effects of climate change on native species as well as crop plants. Here, we provide a toolbox with definitions of key theoretical elements and a synthesis of the current understanding of the molecular and genetic mechanisms underlying plasticity relevant to climate change. By bringing ecological, evolutionary, physiological and molecular perspectives together, we hope to provide clear directives for future research and stimulate cross-disciplinary dialogue on the relevance of phenotypic plasticity under climate change.

Reproductive allocation and resource compensation in male-sterile and hermaphroditic plants of Plantago lanceolata (Plantaginaceae).

Gynodioecy is a breeding system in which hermaphrodites coexist with male steriles. Theoretical models predict that without any compensation in female fitness male steriles will disappear from a population due to their reproductive disadvantage. In the present study I investigated whether male-sterile (MS), partially male-sterile (IN), and hermaphroditic (H) plants of Plantago lanceolata differed in reproductive growth and allocation. Offspring of three interpopulation crosses segregating all three sex morphs were grown under nitrogen-limited conditions in a growth chamber. Independent of the genetic background MS plants attained a higher vegetative and reproductive dry mass and a higher reproductive output than H plants, whereas IN plants had intermediate values. When corrected for the mass of the pollen, the dry mass differences between the sex morphs were much reduced but still present. However, when whole-plant allocation was expressed on the basis of nitrogen, the differences between the sex morphs disappeared. Thus the sex morphs took up similar amounts of nitrogen but distributed them differently. The MS and IN plants used the nitrogen saved by not producing pollen for additional vegetative as well as reproductive growth. The data presented in this study suggest that resource compensation is one of the main mechanisms responsible for the maintenance of MS and IN plants in gynodioecious P. lanceolata.

Evidence for a significant contribution by peroxidase-mediated O2 uptake to root respiration of Brachypodium pinnatum.

This study describes the O2 uptake characteristics of intact roots of Brachypodium pinnatum. In the presence of 25 mM salicylhydroxamic acid (SHAM), concentrations of KCN below 3.5 νM had no effect on the rate of root respiration, whereas in the absence of 25 mM SHAM a significant inhibition of approx. 18% was observed. This indicates that an O2-consuming reaction, not associated with the cytochrome pathway, the alternative pathway or the "residual component", operates in the absence of any inhibitors in roots of B. pinnatum. We demonstrate here that this fourth O2-consuming reaction is mediated by a peroxidase. A peroxidase which catalyzed O2 reduction in the presence of NADH was readily washed from the roots of B. pinnatum. This peroxidase was stimulated by 5 mM SHAM, whereas ascorbic acid, catalase, catechol, gentisic acid, low concentrations potassium cyanide (3.5 µM), sodium azide, sodium sulfide, superoxide dismutase and high concentrations SHAM (25 mM) inhibited this reaction. Except for high concentrations of SHAM and concentrations of KCN higher than approx. 3.5 µM, these effectors could not be used to inhibit the peroxidase-mediated O2 uptake in intact roots of B. pinnatum. Concentrations of SHAM below 10 mM stimulated O2 uptake up to 15% of the control rate, depending on concentration, whereas 25 mM SHAM inhibited O2 uptake by 35%. The stimulation at low concentrations resulted from a SHAM-stimulated peroxidase activity, whereas 25 mM SHAM completely inhibited both the peroxidase-mediated O2 uptake and the activity of the alternative pathway. A method is presented for determining the relative contributions of each of the four O2-consuming reactions, i.e. the cytochrome pathway, the alternative pathway, the "residual component" and the peroxidase-mediated O2 uptake. The peroxidase-mediated O2 uptake contributed 21% to the total rate of oxygen uptake in roots of B. pinnatum, the cytochrome pathway contributed 41%, the alternative pathway 14% and the "residual component" 24%.

A vasopressin and oxytocin containing nucleus in the pig hypothalamus that shows neuronal changes during puberty.

A vasopressin and oxytocin containing nucleus is described for the first time in the pig hypothalamus. It is located near the third ventricle, just dorsal to the suprachiasmatic nucleus, and consists of magnocellular neurons, similar to those of the supraoptic nucleus and paraventricular nucleus. Morphometric analysis of neuronal number, size, density, and volume was performed at four different ages: 1 day, 7 weeks, 16 weeks, and 30 weeks postnatally. No sex difference in these parameters was observed. In this period the volume of the nucleus increased gradually from 6.6 x 10(-3) to 54.2 x 10(-3) mm3. One day after birth 1,215 +/- 191 (mean +/- SEM) neurons were present in the vasopressin and oxytocin containing nucleus, followed by a decrease to 771 +/- 80 neurons at 7 weeks and 697 +/- 116 at 16 weeks. Between 16 and 30 weeks (puberty) there was a dramatic increase in neuron number up to 1,765 +/- 214 neurons. This increase in the number of vasopressin and oxytocin containing neurons in the pig hypothalamus is much later in development than has ever been reported so far.

Prolactin in the developing pig.

Prolactin (PRL) was determined in plasma of fetal pigs from 40 days post coitum (d.p.c.) onwards. Values increased in the last 3 wk of the gestational period and reached 10 ng/ml at term. Immunoreactive lactotropes could be identified as early as 31 d.p.c. Lactotrope development appeared to be biphasic, with no apparent increase in cell number in the 51-75 d.p.c. period. Sex differences in volume percentage of lactotropes and plasma PRL levels were observed in the 71-75 d.p.c. period. Pituitary PRL content increased 20-fold between 95 d.p.c. and term and 10-fold between birth and 6 wk. Volume percentage of lactotropes did not change from birth to 6 wk. In the postnatal period, plasma PRL concentrations were high in the first 1-2 wk and they decreased thereafter. Lower stable values were observed until about 10 wk, when PRL concentrations began to increase. Prepubertal peak levels were observed in the 10- to 16-wk period and a pubertal peak was observed at 20-22 wk. No significant differences between plasma PRL profiles of males and females were observed in the postnatal period. Castrated males, however, had consistently lower plasma PRL concentrations than intact boars. Cryptorchid boars had PRL concentrations comparable to intact boars. The PRL profile, in relation to the pattern of gonadal development, suggests a role for PRL in gonadal development of the pig.

PAP complex: a pitfall in immunocytochemistry of the pig hypothalamus.

Use of rabbit peroxidase-antiperoxidase (PAP) complex in immunostaining of the pig hypothalamus resulted in false positive staining of elements of the supraoptic and paraventricular nuclei due to binding of rabbit PAP complex and rabbit immunoglobulins to these structures. This phenomenon seriously hampers immunohistochemical investigation of the pig hypothalamus. Specific binding of swine immune serum against luteinizing hormone-releasing hormone has been detected with the protein A-biotin-streptavidin method employed on paraffin and vibratome sections of fetal and postnatal pig hypothalamus.

Development of pituitary gonadotropic cells in the pig fetus and the effect of luteinizing hormone-releasing hormone administration.

Pituitary gonadotropic development in fetal pigs has been studied immunocytochemically. From 50 days postcoitum (p.c.) until 90 days p.c. the volume density of luteinizing hormone (LH) immunoreactive cells increased from 0.12% to 0.61% of the glandular mass of the anterior part of the pituitary gland. From 90 to 100 days p.c. a steep increase to 4.5% was found. A single injection of long-acting luteinizing hormone-releasing hormone (LHRH) to the fetus at 75 days p.c. inhibited this developmental pattern. Pulsatile administration of LHRH agonist to fetuses from 70 to 73 days p.c. had no significant effect on the volume density of LH immunoreactive cells, whereas pulsatile administration from 70 to 80 days p.c. and from 95 to 105 days p.c. inhibited the normal increase of this parameter. It is concluded that from 70 days p.c. onwards the development of fetal gonadotropic cells can be manipulated by exogenous LHRH.

Nerve endings in the pulmonary trunk, ductus arteriosus and aorta of intact and decapitated pig fetuses.

Nerve fibres reactive to acetyl-thiocholine, and tissues showing catecholamine fluorescence were examined in the pulmonary trunk, ductus arteriosus and aorta of 28 pig fetuses between 31 and 113 days of gestation (term = 114 +/- 1 days). Eight additional fetuses, which had been decapitated in utero at 40-43 days, were also studied at ages between 51 and 114 days of gestation. Spherical micro-networks of nervous tissue reactive to acetyl-thiocholine are present in the adventitia on the cranial aspect of the pulmonary trunk and ductus arteriosus, between the aorta and pulmonary trunk, and on the caudal aspects of the pulmonary trunk and the pulmonary arteries. These fibres invest spherical clusters of catecholamine containing cells which are well supplied with blood vessels. Nerve fibres which fluoresce are also found in association with these cells. Decapitation in utero does not appear to affect the distribution of morphology of these structures. The observations show that structures are present in the major arteries of the fetal pig which may act as sensory receptors, and that these structures are unaffected by chronic vagotomy of the fetus produced by decapitation early in gestation.

Development of nervous tissue in the heart of the fetal and neonatal pig and the effect of decapitation in utero.

The development and distribution of the nerves in the heart of the pig was studied macroscopically and by light microscopy. Hearts were collected from 86 fetuses between 31 and 114 days of gestation (term = 114 days), from 12 neonatal pigs aged 9 and 20 days and from 6 adult sows of the Dutch Landrace breed. The effect of vagotomy produced by decapitation in utero at 40-43 days was studied in an additional 24 hearts from fetuses aged between 51 and 114 days of gestation. The amount of acetyl-thiocholine reactive fibres increases in the atria, A-V node and ventricles throughout gestation. At every age the amount of nervous tissue is highest in the A-V node and lowest in the ventricles. Hearts from decapitated fetuses have smaller amounts of nerve tissue than those from intact fetuses at every age studied. Ganglia are present in both intact and decapitated fetuses. Fluorescent cells containing catecholamines are observed in hearts from fetuses as young as 35 days gestation. Although fluorescent nerve fibres are rarely seen in hearts at 70 days gestation, more fibres are present near birth and thereafter there appears to be a considerable increase in the number of fibres and in the intensity with which they fluoresce. These results show that there is substantial nerve growth into the heart of the pig during gestation and that catecholamine containing nerve fibres develop later than those reactive to acetyl-thiocholine.