Involvement of polyamines in root development at low temperature in the subantarctic cruciferous species Pringlea antiscorbutica

Polyamine involvement in root development at low temperature was studied in seedlings of Pringlea antiscorbutica R. Br. This unique endemic cruciferous species from the subantarctic zone is subjected to strong environmental constraints and shows high polyamine contents. In the present study, free polyamine levels were modified by inhibitors of polyamine biosynthesis (D-arginine, difluoromethylornithine, cyclohexylammonium, and methylglyoxal-bis-guanylhydrazone) and variations of the endogenous pools were compared to changes in root growth. The arginine decarboxylase pathway, rather than that of ornithine decarboxylase, seemed to play a major role in polyamine synthesis in Pringlea antiscorbutica seedlings. Root, but not shoot, phenotypes were greatly affected by these treatments, which modified polyamine endogenous levels according to their expected effects. A positive correlation was found between agmatine level and growth rate of the primary root. Spermidine and spermine contents also showed positive correlations with primary root growth whereas the putrescine level showed neutral or negative effects on this trait. Free polyamines were therefore found to be differentially involved in the phenotypic plasticity of root architecture. A comparison of developmental effects and physiological concentrations suggested that agmatine and spermine in particular may play a significant role in the control of root development.     

Growth and reproduction of the endemic cruciferous species Pringlea antiscorbutica in Kerguelen Islands

This paper presents the first results from a 7-year monitoring of Pringlea plants established naturally from seed at Kerguelen at two sites with different microenvironmental characteristics. The field growth and reproductive traits of Pringlea are reported for the first time. Pringlea plants grow much faster than was previously believed, attaining around 50 cm diameter in 4 years. The growth pause in winter is short. Pringlea first flowers mainly in its 3rd or 4th year of growth and, as such, this species can be described as an early-flowering perennial. Inter-individual variability for all growth and reproductive parameters was generally higher than inter-site variability. These biological traits are compared to other subantarctic phanerogams and are discussed in terms of adaptation to subantarctic climate and ecological distribution of Pringlea antiscorbutica. Accepted: 27 September 1999     

Carbon metabolism in the subantarctic Kerguelen cabbage Pringlea antiscorbutica R. Br.: environmental controls over carbohydrates and proline contents and relation to phenology

The subantarctic Brassicaceae Pringlea antiscorbutica R. Br. (Kerguelen cabbage) was used as a model to study the physiological adaptations of higher plants to the subantarctic environment. ¹³C-nuclear magnetic resonance permitted, in combination with biochemical methods, the identification and quantification of the major solutes in leaves, stem and roots. As characterized in many Brassicaceae, proline was a major solute in all organs of the plants, and its accumulation was mainly controlled by salt stress rather than temperature. Glucose was the major soluble sugar in the leaves, whilst sucrose and starch accumulated in stems and roots. Over a period of 1 year we found strong correlations between (i) glucose content in leaves and irradiance, and (ii) starch content in non-photosynthetic organs and air temperature. The pattern of carbohydrate accumulation indirectly indicated that photosynthetis was sustained throughout the year, even during cold days when the temperature remained near 0 °C. This is consistent with the direct gas exchange measurements showing that photosynthetic capacity is mainly influenced by irradiance and weakly by temperature. Taken together, these characteristics demonstrated that the growth and development cycle occurs without a period of dormancy.     

Subcellular compartmentation of proline in the leaves of the subantarctic Kerguelen cabbage Pringlea antiscorbutica R. Br. In vivo13C-NMR study

Proline is one of the major solutes accumulated upon salt stress in leaves, stem and roots of the subantarctic Brassicaceae Pringlea antiscorbutica R. Br. (Kerguelen cabbage). Using in vivo ¹³C-NMR techniques, it was possible for the first time to visualize the subcellular compartmentation of proline between cytoplasmic and vacuolar compartments in Pringlea leaves. We observed that this osmolyte accumulated at a 2–3 times higher concentration in the cytoplasm than in the vacuole.     

Breeding system of the subantarctic plant species Pringlea antiscorbutica R. Br. and search for potential insect pollinators in the Kerguelen Islands

The reproductive biology of Pringlea antiscorbutica R. Br. (Brassicaceae), an endemic species from the southern Indian Ocean islands, is investigated here. Controlled crosses were performed between plants grown in a common garden on the Kerguelen Islands. Searching for potential insect pollinators, we investigated the flying ability of all insects known from the Kerguelen Islands. We showed evidence for self-compatibility, low pollen limitation in the absence of a biotic pollen vector and slight selfing depression in P. antiscorbutica. The insects present on the islands are mostly wingless and are not likely to be efficient pollinators of this species. Since P. antiscorbutica shows morphological adaptations to insect pollination despite the absence of pollinators in its present range, we conclude that this species recently evolved from insect pollination to wind and/or autonomous pollination.     

High flexibility in growth and polyamine composition of the crucifer Pringlea antiscorbutica in relation to environmental conditions

Although polyamine (PA) metabolism in plants responds to abiotic stresses, few studies have investigated this response in plants under natural conditions. Using high-performance liquid chromatography, we studied the amine composition of the subantarctic crucifer Pringlea antiscorbutica R. Br. both in the field (Crozet and Kerguelen Islands) and under controlled temperatures in the laboratory. Plants collected from different sites showed a large variability of amine composition and contents. The aliphatic and acetylated amine composition of leaves allowed to statistically identify two groups. The first group, composed of Kerguelen coastal plants, was characterized by high levels of acetylspermidine and acetylspermine, showing the presence in P. antiscorbutica of this uncommon regulation pathway. PA acetylation may be induced in P. antiscorbutica under conditions of low water availability. The second group, composed of plants from Kerguelen altitude sites and plants from Crozet sites, showed high levels of free spermidine (Spd). In these plants, the ratio Spd/putrescine did not positively correlate with plant size as was found in developmental studies, suggesting that free Spd may be devoted to other aims, such as cold tolerance. Some differences between Crozet and Kerguelen plant responses suggested the possibility that different regulations of amine metabolism could take place in plants from these two islands. Agmatine accumulation pattern was diverse and suggested this amine to be sensitive to combinations of environmental factors. Studies on amine variation patterns in P. antiscorbutica provide insights into the roles of rarely reported amines, such as acetylated amines, in plant metabolic adaptation to abiotic stresses.     

Phylogeny and colonization history of Pringlea antiscorbutica (Brassicaceae), an emblematic endemic from the South Indian Ocean Province

The origins and evolution of sub-Antarctic island floras are not well understood. In particular there is uncertainty about the ages of the contemporary floras and the ultimate origins of the lineages they contain. Pringlea R. Br. (Brassicaceae) is a monotypic genus endemic to four sub-Antarctic island groups in the southern Indian Ocean. Here we used sequences from both the chloroplast and nuclear genomes to examine the phylogenetic position of this enigmatic genus. Our analyses confirm that Pringlea falls within the tribe Thelypodieae and provide a preliminary view of its relationships within the group. Divergence time estimates and ancestral area reconstructions imply Pringlea diverged from a South American ancestor ～5Myr ago. It remains unclear whether the ancestor of Pringlea dispersed directly to the South Indian Ocean Province (SIOP) or used Antarctica as a stepping-stone; what is clear, however, is that following arrival in the SIOP several additional long-distance dispersal events must be inferred to explain the current distribution of this species. Our analyses also suggest that although Pringlea is likely to have inherited cold tolerance from its closest relatives, the distinctive morphology of this species evolved only after it split from the South American lineage. More generally, our results lend support to the hypothesis that angiosperms persisted on the sub-Antarctic islands throughout the Pliocene and Pleistocene. Taken together with evidence from other sub-Antarctic island plant groups, they suggest the extant flora of sub-Antarctic is likely to have been assembled over a broad time period and from lineages with distinctive biogeographic histories.     

Accumulation of organic and inorganic solutes in the subantarctic cruciferous species Pringlea antiscorbutica in response to saline and cold stresses

Pringlea antiscorbutica R. Br., a subantarctic endemic cruciferous species, is endangered in its natural sites by several ecological changes. This species is tolerant to salinity and a permanent cold temperature on Kerguelen and Crozet Islands. We attempted the investigation of regulating mechanisms of osmotic adjustment in this species. ¹³C NMR analyses of water-soluble compounds from leaves collected from the field revealed glucose and proline to be the main accumulated organic solutes. Colorimetric determinations in these samples showed that proline and soluble carbohydrates were present at remarkably high levels. When young plants were cultivated in growth chambers they showed a good resistance to cold and medium resistance to saline conditions. High levels of soluble carbohydrates were present in all situations. Proline was accumulated in response to a saline and a cold treatment. The quantitative variations of the pool of proline in response to saline treatments were rapid and important. The adaptive value of these responses of organic solutes in the tolerance of Pringlea antiscorbutica to various stresses is discussed. Received: 16 June 1997 / Accepted: 5 May 1998     

Effects of salinity and flooding on seedlings of cabbage palm (Sabal palmetto)

Sabal palmetto (Walt.) Lodd. ex Schultes (cabbage palm) dominates the coastal limit of many forests in North Florida and Georgia, United States. Changes in saltwater flooding due to sea level rise have been credicted with pushing the coastal limit of cabbage palms inland, eliminating regeneration before causing death of mature trees. Localized freshwater discharge along the coast causes different forest stands to experience tidal flooding with waters that differ in salinity. To elucidate the effect of such variation on regeneration failure under tidal flooding, we examined relative effects of flooding and salinity on the performance of cabbage palm seedlings. We examined the relationship between seedling establishment and degree of tidal inundation in the field, compared the ability of seedlings to withstand tidal flooding at two coastal sites that differed in tidal water salinity, and investigated the physiological responses of cabbage palm seedlings to salinity and flooding in a factorial greenhouse experiment. Seedling survival was inversely correlated with depth and frequency of tidal flooding. Survival of seedlings at a coastal site flooded by waters low in salinity [c. 3 parts per thousand (ppt)] was greater than that at a site flooded by waters higher in salinity (up to 23 ppt). Greenhouse experiments revealed that leaves of seedlings in pots flushed twice daily with salt solutions of 0 ppt and 8 ppt exhibited little difference in midmorning net CO₂ assimilation rates; those flushed with solutions of 15 ppt and 22 ppt, in contrast, had such low rates that they could not be detected. Net CO₂ assimilation rates also declined with increasing salinity for seedlings in pots that were continuously inundated. Continuous root zone inundation appeared to ameliorate effects of salinity on photosynthesis, presumably due to increased salt concentrations and possibly water deficits in periodically flushed pots. Such problems associated with periodic flushing by salt water may play a role in the mortality of cabbage palm seedlings in the field. The salinity range in which plant performance plummeted in the greenhouse was consistent with the salinity difference found between our two coastal study sites, suggesting that variation in tidal water salinity along the coast plays an important role in the ability of cabbage palm seedlings to withstand tidal flooding.     

The effects of root temperature and Ca supply on the growth and transpiration of cotton seedlings (Gossypium hirsutum,L.)

Summary Cotton seedlings were germinated in either tap water or a CaSO₄ solution and then grown for two days in nutrient solutions containing 0.1, 1.0, or 10.0 me Ca/1. They were then transferred to cultures having the same Ca variables and the roots subjected to temperatures of 26, 18, 15 and 12°C for four days. The fresh weight of all plant parts and leaf area increased with increasing root temperatures and with increasing Ca levels, the effect of Ca being most pronounced at the higher temperatures. The dry weight was increased by increasing root temperature; the effect of Ca was rather small. Water use increased with increasing temperature and was higher for the lowest Ca level than for the higher levels. The results emphasize the overriding effect of low temperature on water uptake. Calcium had little effect on the growth depression resulting from low root temperatures. Joint contribution from the Agronomy Department, Mississippi Agricultural Experiment Station, State College and the Soil Science Department, North Carolina Agricultural Experiment Station, Raleigh. Published with the approval of the Directors of Research as Journal Contribution No.1530 and Paper No.2389 of the Journal Series, respectively. This is a report of research conducted while the author was on sabbatical leave at North Carolina State University, February 1–September 10, 1965.     

Involvement of photobleaching and inhibition of protochlorophyll(ide) accumulation in tentoxin effects on greening of mung bean seedlings

Several types of evidence indicate that tentoxin-caused reduction of chlorophyll accumulation in greening primary leaves of mung bean [ Vigna radiata (L.) Wilczek cv. Berken] is due to both photobleaching and decreased protochlorophyll(ide) synthesis. Greening was greater under dim (2.5 μmol m_-2 s_-1) far-red or white light than under bright (180 to 200 μmol m_-2 s_-1) white light in tentoxin-treated tissues, whereas there was a positive correlation between fluence rate and greening in control tissues. Under continuous white light (100 μmol m_-2 s_-1) chorophyll(ide) accumulation was slower in tentoxin-treated than in control tissues. This was caused by greater photobleaching of newly formed chlorophyll(ide), as well as by decreased protochlorophyll(ide) synthesis. Photobleaching did not affect protochlorophyll(ide) synthesis in control or tentoxin-treated tissues. Chlorophyll(ide) was less stable in tentoxin-treated than in control tissues during a 24 h period of darkness. Plastids of tentoxin-treated tissues had all of the chlorophyll-proteins of control plants. Etioplasts of tentoxin-treated plants contained normal galactolipid contents, but galactolipids in these plants were greatly reduced in white light. Reduced chlorophyll accumulation caused by tentoxin is apparently the result of both photodestruction and of reduced synthesis of chlorophyll.     

Effect of low temperature on ethanolic fermentation in rice seedlings

Rice seedlings (Oryza sativa L.) were incubated at 5-30 degrees C for 48 h and the effect of temperature on ethanolic fermentation in the seedlings was investigated in terms of low-temperature adaptation. Activities of alcohol dehydrogenase (ADH, EC 1.1.1.1) and pyruvate decarboxylase (PDC, EC 4.1.1.1) in roots and shoots of the seedlings were low at temperatures of 20-30 degrees C, whereas temperatures of 5, 7.5 and 10 degrees C significantly increased ADH and PDC activities in the roots and shoots. Temperatures of 5-10 degrees C also increased ethanol concentrations in the roots and shoots. The ethanol concentrations in the roots and shoots at 7.5 degrees C were 16- and 12-times greater than those in the roots and shoots at 25 degrees C, respectively. These results indicate that low temperatures (5-10 degrees C) induced ethanolic fermentation in the roots and shoots of the seedlings. Ethanol is known to prevent lipid degradation in plant membrane, and increased membrane-lipid fluidization. In addition, an ADH inhibitor, 4-methylpyrazole, decreased low-temperature tolerance in roots and shoots of rice seedlings and this reduction in the tolerance was recovered by exogenous applied ethanol. Therefore, production of ethanol by ethanolic fermentation may lead to low-temperature adaptation in rice plants by altering the physical properties of membrane lipids.     

Involvement of polyamines in the control of fruitlet physiological abscission in grapevine (Vitis vinifera)

The potential contribution of polyamines (PAs) in the regulation of physiologically induced fruitlet abscission was investigated in cuttings from two cultivars of Vitis vinifera L., Pinot noir (PN) and Merlot (MRT). Abscission was higher in MRT than in PN and was preceded by a decrease in free PA levels. This decline was more pronounced in inflorescences than in leaves of the sensitive cultivar. Soluble conjugated PA showed an opposite trend in both cultivars. This suggests a cause-effect relationship between free and/or conjugated PA levels in floral organs and susceptibility to abscission. Spermidine (Spd), but not putrescine (Put) or diaminopropane, supplied at 0.5–1 m M to the nutritive medium prior to the anthesis, increased free and conjugated PA levels in the inflorescences and markedly inhibited abscission. α -Difluoromethylarginine, an inhibitor of arginine decarboxylase, but not α -difluoromethylornithine, an inhibitor of ornithine decarboxylase, lowered PA levels and increased abscission. Treatment with cyclohexylamine or β -hydroxyethylhydrazine as potent inhibitors of Spd synthase and PA oxidases, respectively, reduced the Spd and/or spermine levels and enhanced free Put in the inflorescences, inducing an increased abscission of floral organs shortly after anthesis. These data suggest that PAs, particularly Spd, could be involved in the regulation of grapevine fruitlet physiological abscission.     

Temporal organization of chilling resistance in cotton seedlings: effects of low temperature and relative humidity

The effect of temperature and relative humidity (RH) on the time course of the rhythmic endogenous changes of chilling resistance was studied in cotton (Gossypium hirsutum L. cv. Deltapine 50) seedlings grown under light-dark cycles of 12:12 h. The resistant phase to 5° C, 85% RH lasted during most of the dark period while to 5° C, 100% RH it was longer and extended into the last half of the light period because a transient phase advance occurred when chilling started at the middle of the light period. Seedlings acclimated by low temperature were resistant throughout the light-dark cycle. A treatment with 100% RH before chilling to acclimated seedlings introduced a sensitive phase that corresponded to that of non-acclimated seedlings. In non-acclimated seedlings, this treatment decreased the resistance but the basic pattern of the rhythm was sustained. Changes in chilling resistance were analyzed under fluctuating temperatures and RHs, and explained taking into consideration the functioning of the circadian clock and environmental induction of resistance.Abbreviations CR chilling resistance - LDC light-dark cycle of 24 h - RH relative humidity     

Stimulation of androgenesis in white cabbage (Brassica oleracea var. capitata) anthers by low temperature and anther dissection

Anther culture was performed on two local cultivars, Ljubljansko and Varadinsko, and the F₁ cv. Krautman (Bejo-Zaden). The effects on androgenesis of hot and cold temperature treatments and different dissections of anthers were evaluated. In contrast to cv. Krautman, cvs. Ljubljansko and Varadinsko produced more embryos after cold pretreatment of flower buds (4°C, 48 h) than after standard treatment (35°C, 24h). Simultaneous cutting of the anther tip and removal of the filament gave the best results in comparison to other tested dissections. Microscopical observations of sectioned anthers revealed enhanced embryo development near the cut ends of the anthers. Ploidy analysis revealed the presence of haploids among embryos resulting from cold treatment (4°C, 48 h), treatment at elevated temperature (35°C, 24 h), and among embryos resulting from dissections of anther tips.     

低镁胁迫对低温下黄瓜幼苗光合特性和抗氧化系统的影响

以‘津优3号’黄瓜幼苗为试材,以Hoagland全营养液处理为对照(CK),研究低镁(30％Mg)胁迫对低温(昼/夜温度12℃/8℃)下黄瓜幼苗光合特性和抗氧化系统的影响.结果表明:低温下30％Mg处理黄瓜幼苗叶片Mg含量显著低于CK,而根中Mg含量与CK差异不显著.随着低温胁迫时间的延长,黄瓜幼苗叶片的叶绿素含量、净光合速率(Pn)、气孔导度(gs)和羧化效率(CE)逐渐降低,胞间CO2浓度(Ci)趋于升高.与CK相比,低温下30％ Mg处理叶片叶绿素含量、Pn、gs和CE显著降低,C-变化不大,叶绿体膜损伤严重,叶绿体数、基粒数和片层数较少,淀粉粒数增加,淀粉粒较长,丙二醛(MDA)含量升高,而超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性降低.可见,低温下镁运输受阻引起的缺镁是叶片失绿的主要原因;低温引起Pn降低的主要原因是非气孔限制,低镁胁迫会加大低温对黄瓜Pn的影响,而由此引起的Pn降低的主要原因是气孔限制.