Proteomic changes of Citrus roots in response to long-term manganese toxicity

Key message

Fifty-three and thirty-nine differentially expressed protein spots were isolated from Mn-toxic Citrus sinensis and Citrus grandis roots, respectively. Mn-toxicity-induced changes in protein profiles greatly differed between the two species.

Abstract

Limited information is available on the manganese (Mn)-toxicity-responsive proteins in plant roots. ‘Sour pummelo’ (Citrus grandis) and ‘Xuegan’ (Citrus sinensis) seedlings were irrigated for 17 weeks with 2 (control) or 600 μM (Mn-toxic) MnSO₄. C. sinensis displayed more tolerance to Mn-toxicity than C. grandis, which may be related to more Mn accumulation in roots and less Mn distribution in shoots. Using two-dimensional electrophoresis (2-DE), we isolated 11 up-regulated and 42 down-regulated protein spots from Mn-toxic C. sinensis roots, and 25 up-regulated and 14 down-regulated protein spots from Mn-toxic C. grandis roots. This indicated more metabolic flexibility in C. sinensis roots, thus contributing to the Mn-tolerance of C. sinensis. According to the biological functional properties, these differentially expressed proteins in the two species were classified into the following categories: protein metabolism, nucleic acid metabolism, carbohydrate and energy metabolism, stress responses, cell wall and cytoskeleton, cell transport, signal transduction and fatty acid metabolism. Under Mn-toxicity, proteins involved in nucleic acid metabolism, glycolysis and cell transport were up-regulated in nontolerant C. grandis roots, and down-regulated in tolerant C. sinensis roots. The notable down-regulation of proteins in Mn-toxic C. sinensis roots with less accumulation of carbohydrates may provide an advantage to the net carbon balance by lowering related metabolic processes, and enhancing the Mn-tolerance of C. sinensis. To conclude, there are many important differences in Mn-toxicity-induced changes in protein profiles and metabolic responses between the two species.     

New taxa in Pectis (Compositae: Pectidinae) from Mexico and the southwestern United States

Pectis minutiflora,P. papposa var.grandis, andP. purpurea var.sonorae are described as new. A new combination,P. purpurea var.lancifolia also is included. Chromosome counts (n = 12) are reported forP. papposa var.grandis andP. purpurea var.sonorae.     

Roles of rootstocks and scions in aluminum-tolerance of <Emphasis Type="Italic">Citrus</Emphasis>

Four scion-rootstock combination [i.e., X/X and X/SP, ‘Xuegan’ (Citrus sinensis) grafted on ‘Xugan’ and ‘Sour pummelo’ (Citrus grandis), respectively, and SP/X and SP/SP, ‘Sour pummelo’ grafted on ‘Xuegan’ and ‘Sour pummelo’, respectively] plants were treated for 18 weeks with 0 (?Al) or 1.2 mM AlCl₃·6H₂O (+Al). Thereafter, leaf, stem and root concentrations of phosphorus and aluminum (Al), leaf and root levels of organic acids (OAs), Al-induced release of OA anions (i.e., malate and citrate), photosynthesis and chlorophyll a fluorescence (OJIP) transients were measured. Al-induced decrease of photosynthesis and damage of photosynthetic electron transport chain were less pronounced in X/X and X/SP leaves than in SP/SP and SP/X leaves, which might be related with the higher Al-induced root efflux of OA anions and leaf P concentration. C. sinensis rootstock alleviated the influences of Al-toxicity on leaf photosynthetic electron transport chain by enhancing Al-induced release of root OA anions, hence lessening Al-induced photosynthesis inhibition in SP/X plants, while the reverse was the case for C. grandis rootstock in X/SP plants. In conclusion, the tolerance of grafted Citrus plants to Al depends on the scion as well as rootstock genotype, and the scion-rootstock interaction.     

New species and new combinations in the sematophyllum subpinnatum complex (Sematophyllaceae)

Three new tropical American species ofSematophyllum are described with relatively broad leaves and short upper leaf cells:S. cubense, S. cataractae, andS. oedophysidium. The Central AmericaS. apaloblastum andS. barnesii are new combinations.Sematophyllum galipense, S. amnigenum, andS. cochleatum are briefly discussed.     

Converging patterns of vertical variability in leaf morphology and nitrogen across seven Eucalyptus plantations in Brazil and Hawaii,USA

Key message

Across sites in Brazil and Hawaii, LMA and N _mass were strongly correlated with height and shade index, respectively, which may help simplify canopy function modeling of Eucalyptus plantations.

Abstract

Within tree canopies, leaf mass per area (LMA) and leaf nitrogen per unit area (N _area) commonly increase with height. Previous research has suggested that these patterns occur as a strategy to optimize carbon gain by allocating available resources to upper canopy leaves that are exposed to greater light availability. We tested three hypotheses about the influences of height, shade index (a proxy for light), and stand age on LMA and leaf nitrogen for even-aged Eucalyptus saligna and Eucalyptus grandis × urophylla plantations in Brazil and Hawaii, USA, spanning most of the environmental conditions found across 19.6 million ha of Eucalyptus spp. plantations around the world. Shade index was developed by incorporating canopy depth (inner-crown shading) and a tree height ratio relative to neighbor trees (shading from other trees). Across all sites and ages, leaf height accounted for 45 % of the variation in LMA, whereas shade index accounted for only 6 %. A combination of both factors was slightly better in accounting for LMA variation than height alone. LMA–height relationships among sites were strongest under greater light availability and in older stands. Leaf nitrogen per unit mass (N _mass) consistently decreased with shade index, whereas N _area showed no consistent pattern with height or shade index. These relationships indicate that N _mass is primarily driven by light, while height is the primary driver for LMA. The general relationships between LMA and leaf N _mass across all sites may simplify canopy function modeling of E. saligna and E. grandis × urophylla plantations.     

A new wild type of Citron (Citrus medica L., Rutaceae) identified through morphology and psbM-trnD spacer region of chloroplast DNA

Key message

This study showed that Themachhi is genetically very close to C. medica and consistently grouped with the typical citron in all the phylogenetic trees; hence revealed the identity of ‘Themachhi’ as a natural wild type of C. medica.

Abstract

Taxonomic identity and well-supported phylogenetic relationships of doubtful and taxonomically uncertain taxa are highly requisite for developing crop improvement programmes and suitable conservation strategies. In Citrus, several natural varieties and hybrids of ambiguous origin have been existing due to several factors such as frequent intergeneric and interspecific hybridization, apomixis, bud mutations, polyploidy and diverse eco-geographical distribution. In Northeast India, an important unidentified type of Citrus, i.e. ‘Themachhi’ is found naturally in Garo Hills, Meghalaya, which is morphologically very close to C. medica and C. limon. The sequences of chloroplast non-coding region psbM-trnD of 16 accessions of Citrus were utilized to confirm the taxonomic identity of ‘Themachhi’ and also to infer its phylogenetic affinities to related taxa. The nucleotide frequencies were 32.86 % for A and T and 17.4 % for C and G. Pairwise genetic divergence between the accessions of Citrus ranged from 0.002 to 0.036 with an average of 0.017. ‘Themachhi’ showed least genetic divergence with C. medica (0.013) and highest divergence with C. limon (average 0.024). All the accessions of Citrus were grouped into three main clusters (C. medica, C. reticulata and C. maxima clusters) in both the MP and ML trees with high bootstrap value (>60 %). In the MP and ML trees, both accessions of ‘Themachhi’ were closely attached with C. medica cluster with high bootstrap value (98 %). Close genetic affinity and consistently grouping of “Themachhi” with typical citron (C. medica) in both the phylogenetic trees revealed the identity of ‘Themachhi’ as a natural wild type of C. medica.     

A new variety of coreopsis mutica (Compositae) from Mexico

Coreopsis mutica var.simplicifolia is described from the state of Guanajuato in Mexico. The new variety is distinguishable by its simple lanceolate to linear-lanceolate leaves. In addition, var.simplicifolia is al lopatric with other C.mutica and has a distinctive leaf flavonoid chemistry. The chromosome number of the variety is 2n = 56.     

Within-canopy variation in photosynthetic capacity,SLA and foliar N in temperate broad-leaved trees with contrasting shade tolerance

Key message

The relative shade tolerance of T. cordata , F. sylvatica , and C. betulus in mature stands is based on different species-specific carbon and nitrogen allocation patterns.

Abstract

The leaf morphology and photosynthetic capacity of trees are remarkably plastic in response to intra-canopy light gradients. While most studies examined seedlings, it is not well understood how plasticity differs in mature trees among species with contrasting shade tolerance. We studied light-saturated net photosynthesis (A _max), maximum carboxylation rate (V _cmax), electron transport capacity (J _max) and leaf dark respiration (R _d) along natural light gradients in the canopies of 26 adult trees of five broad-leaved tree species in a mixed temperate old-growth forest (Fraxinus excelsior, Acer pseudoplatanus, Carpinus betulus, Tilia cordata and Fagus sylvatica), representing a sequence from moderately light-demanding to highly shade-tolerant species. We searched for species differences in the dependence of photosynthetic capacity on relative irradiance (RI), specific leaf area (SLA) and nitrogen per leaf area (N _a ). The three shade-tolerant species (C. betulus, T. cordata, F. sylvatica) differed from the two more light-demanding species by the formation of shade leaves with particularly high SLA but relatively low N _a and consequently lower area-based A _max, and a generally higher leaf morphological and functional plasticity across the canopy. Sun leaf morphology and physiology were more similar among the two groups. The three shade-tolerant species differed in their shade acclimation strategies which are primarily determined by the species’ plasticity in SLA. Under low light, T. cordata and F. sylvatica increased SLA, mass-based foliar N and leaf size, while C. betulus increased solely SLA exhibiting only low intra-crown plasticity in leaf morphology and N allocation patterns. This study with mature trees adds to our understanding of tree species differences in shade acclimation strategies under the natural conditions of a mixed old-growth forest.     

Xcc-facilitated agroinfiltration of citrus leaves: a tool for rapid functional analysis of transgenes in citrus leaves

Key message

Xanthomonas citri subsp. citri pretreatment before agroinfiltration could significantly promote transient expression in citrus leaves which were previously recalcitrant to agroinfiltration.

Abstract

Transient expression via agroinfiltration is widely used in biotechnology but remains problematic in many economically important plants. Xanthomonas citri subsp. citri (Xcc)-facilitated agroinfiltration was employed to promote transient protein expression in Valencia sweet orange leaves, which are recalcitrant to agroinfiltration. However, it is unclear whether Xcc-facilitated agroinfiltration has broad application, i.e., whether Xcc-facilitated agroinfiltration could be used on other citrus varieties. In addition, we intended to investigate whether Xcc-facilitated agroinfiltration could be used to hasten transgene function assays, e.g., Cre/lox system and Cas9/sgRNA system. In this report, Xcc-facilitated agroinfiltration was further exploited to enhance β-glucuronidase (GUS) expression in five citrus varieties. Xcc-facilitated agroinfiltration also significantly increased GFP expression in six citrus varieties tested. Both GUS and GFP assays indicated that Xcc-facilitated agroinfiltration had the best performance in grapefruit. After Xcc-facilitated agroinfiltration was carried out in grapefruit, protoplast analysis of the transformed cells indicated that there were more than 20 % leaf cells expressing GFP. In grapefruit, usefulness of Xcc-facilitated agroinfiltration was assayed in three case studies: (1) fast functional analysis of Cre/lox system, (2) the heat shock regulation of HSP70B promoter derived from Arabidopsis, and (3) Cas9/sgRNA-mediated genome modification.     

Salicylic acid induces physiological and biochemical changes in Torreya grandis cv. Merrillii seedlings under drought stress

Key message

SA treatment effectively ameliorated the negative effect of moderate drought stress on T. grandis Seedlings through increasing the water content, Pn, proline content, antioxidant enzymes activity and reducing MDA.

Abstract

Water availability is one of the most critical factors that limits the growth and development of plants. Salicylic acid (SA) is an important signal molecule that modulates plant responses to abiotic stress. To elucidate the regulating mechanism of exogenous SA on Torreya grandis cv. Merrillii under different water stresses, a pot experiment was conducted in a greenhouse. Exposure of T. grandis seedlings to drought conditions resulted in reduced growth rate that was associated with a decline in water content and CO₂ assimilation. Foliar application of SA effectively increased the water content, net CO₂ assimilation rate, proline content and antioxidant enzymes activity in the plants, which helped T. grandis to acclimate to moderate drought stress and increase the shoot dry matter. However, when the plants were under severe drought stress, the relative water content and CO₂ assimilation in the SA-treated plants were significantly lower than those in the control plants. Therefore, our results indicated that SA can effectively ameliorate the negative effect of moderate drought stress on T. grandis seedling growth.     

Purple-leaved Ficus lyrata plants produced by overexpressing a grapevine VvMybA1 gene

Key message

This study established an efficient method of regenerating plants of Ficus lyrata and producing purple-leaved F. lyrata plants through genetic transformation using a VvMybA1 gene of grapevine.

Abstract

Ficus lyrata, a species with unique violin- or guitar-shaped leaves, was regenerated from leaf-derived calli cultured on Murashige and Skoog (MS) basal medium supplemented with 4.5 μM N-phenyl-N’-1, 2, 3-thiadiazol-5-yl urea (TDZ) and 0.5 μM α-naphthalene acetic acid (NAA). Leaf discs were inoculated with Agrobacterium tumefaciens strain EHA 105 harboring a binary vector DEAT that contains the VvMybA1 gene and neomycin phosphotransferase (npt II) gene and subsequently cultured on the established regeneration medium supplemented with 100 mg l^?1 kanamycin. Results showed that 87.5 % of the leaf discs produced kanamycin-resistant callus, and 68.8 % of them produced adventitious shoots. Transgenic plants with three leaf colors including green, green-purple, and purple were produced. Regular and quantitative real-time PCR analyses confirmed the integration of transgenes into the host genome. Semi-quantitative RT-PCR analysis indicated that the VvMybA1 gene was responsible for the purple-colored phenotype. Purple-leaved plants with strong color stability grew vigorously in a greenhouse. This study illustrated the feasibility of using a genetically engineered VvMybA1 gene for drastic modification of leaf color of an important woody ornamental plant.     

A new variety of cardamine angustata (cruciferae) from the Ouachita Mountains of Arkansas

Cardamine angustata var.ouachitana E. B. Smith, a new variety differing fromC. angustata var.angustata in absolutely glabrous leaf segments, is described from the Ouachita Mountains of Arkansas.     

A novel insight into the structure of amphivasal secondary bundles on the example of Dracaena draco L. stem

Key message

Pattern of tracheids found along the bundles extends understanding of their cross - sectional anatomy and sheds a new light on the issue of radial transport in monocotyledons with secondary growth.

Abstract

Secondary growth of Dracaena draco L. stem is connected with the formation of amphivasal vascular bundles in which a centrally located phloem is surrounded by a ring of xylem cells (tracheids). However, as visible in a single transverse section, there is a tendency towards variation among the secondary bundles from such with a xylem ring to ones in which the tracheids do not completely surround the phloem, i.e., are separated by vascular parenchyma cells. We aimed to elucidate the cross-sectional anatomy of amphivasal secondary bundles using the method of serial sectioning (with sections 3 μm thick), which allowed us to follow very precisely the bundle structure along its length. The analysis revealed that the xylem arrangement in these bundles depends on the position of a section in the bundle path. Each amphivasal bundle is composed of sectors where tracheids form a ring, as well as of such where tracheids are separated by vascular parenchyma cells. We hypothesize that this structure of amphivasal vascular bundles facilitates radial transport of assimilates to the sink tissues. The result of the anatomical analysis is discussed in a physiological context.     

Function and structure of leaves contributing to increasing water storage with height in the tallest <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> trees of Japan

Key message

In Cryptomeria japonica , transfusion tissue in leaves may have functions of water storage and supply, which could compensate for hydraulic constraints with increasing height.

Abstract

The tallest trees of Cryptomeria japonica occur in climatic regions similar to the world’s tallest trees. We hypothesized that tall C. japonica trees would have evolved adaptive mechanisms to overcome height growth limitation. Here, we focused on foliar water storage, a mechanism recently discovered in Sequoia sempervirens. In C. japonica, leaf water potential at turgor loss did not change with height or light availability, while leaf hydraulic capacitance and succulence (water content per leaf surface area) increased, suggesting hydraulic compensation. Plasticity of leaf morphology could contribute to avoiding negative effects of height on photosynthesis. We also focused on the structure and function of transfusion tissue in leaves and its role in water storage and supply. Cross-sectional area of transfusion tissue increased with height, whereas that of xylem was constant. We confirmed that water flowed from vascular bundle to mesophyll via the transfusion tissue. Cryo-scanning electron microscopy images of leaf cross sections showed that transfusion cells were flattened, but not fully dehydrated when leaf water potential decreased in situ and by experimental dehydration, and cell deformation was more marked for treetop leaves than for lower-crown leaves. The shape of transfusion cells recovered at predawn as well as after experimental rehydration. As in S. sempervirens, transfusion tissue of C. japonica may function as a hydraulic buffer, absorbing and releasing water according to leaf water status. Anatomical and hydraulic properties contributing to foliar water storage may be an adaptive mechanism acquired by tall Cupressaceae trees to overcome the hydraulic constraints on physiological function with increasing height.

     

Overexpression of Camellia sinensis H1 histone gene confers abiotic stress tolerance in transgenic tobacco

Key message

Overexpression of CsHis in tobacco promoted chromatin condensation, but did not affect the phenotype. It also conferred tolerance to low-temperature, high-salinity, ABA, drought and oxidative stress in transgenic tobacco.

Abstract

H1 histone, as a major structural protein of higher-order chromatin, is associated with stress responses in plants. Here, we describe the functions of the Camellia sinensis H1 Histone gene (CsHis) to illustrate its roles in plant responses to stresses. Subcellular localization and prokaryotic expression assays showed that the CsHis protein is localized in the nucleus, and its molecular size is approximately 22.5 kD. The expression levels of CsHis in C. sinensis leaves under various conditions were investigated by qRT-PCR, and the results indicated that CsHis was strongly induced by various abiotic stresses such as low-temperature, high-salinity, ABA, drought and oxidative stress. Overexpression of CsHis in tobacco (Nicotiana tabacum) promoted chromatin condensation, while there were almost no changes in the growth and development of transgenic tobacco plants. Phylogenetic analysis showed that CsHis belongs to the H1C and H1D variants of H1 histones, which are stress-induced variants and not the key variants required for growth and development. Stress tolerance analysis indicated that the transgenic tobacco plants exhibited higher tolerance than the WT plants upon exposure to various abiotic stresses; the transgenic plants displayed reduced wilting and senescence and exhibited greater net photosynthetic rate (Pn), stomatal conductance (Gs) and maximal photochemical efficiency (Fv/Fm) values. All the above results suggest that CsHis is a stress-induced gene and that its overexpression improves the tolerance to various abiotic stresses in the transgenic tobacco plants, possibly through the maintenance of photosynthetic efficiency.     

Stable carbon and nitrogen isotope ratios of Eucalyptus and Acacia species along a seasonal rainfall gradient in Western Australia

Key message

Eucalyptus and Acacia species were surprisingly similar with respect to variations in δ ¹³ C, δ ¹⁵ N. Both genera respond with speciation and associated changes in leaf structure to drought.

Abstract

Stable carbon and nitrogen isotope ratios (δ¹³C and δ¹⁵N) in leaves of eucalypts (Corymbia and Eucalyptus) and Acacia (and some additional Fabaceae) species were investigated together with specific leaf area (SLA), leaf nitrogen (N) and leaf phosphorous (P) concentration along a north–south transect through Western Australia covering winter- and summer-dominated rainfall between 100 and 1,200 mm annually. We investigated 62 eucalypts and 78 woody Fabaceae species, mainly of the genus Acacia. Leaf δ¹³C values of Eucalyptus and Acacia species generally increased linearly with latitude from ?29.5 ± 1.3 ‰ in the summer-dominated rainfall zone (15°S–18°S) to about ?25.7 ± 1.1 ‰ in the winter-dominated rainfall zone (29°S–31°S). δ¹⁵N increased initially with southern latitudes (0.5 ± 1.6 ‰ at 15°S; 5.8 ± 3.3 ‰ at 24–29°S) but decreased again further South (4.6 ± 3.5 ‰ at 31°S). The variation in δ¹³C and δ¹⁵N was probably due to speciation of Eucalyptus and Acacia into very local populations. There were no species that were distributed over the whole sampling area. The variation in leaf traits was larger between species than within species. Average nitrogen concentrations were 11.9 ± 1.05 mg g^?1 in Eucalyptus, and were 18.7 ± 4.1 mg g^?1 in Acacia. Even though the average nitrogen concentration was higher in Acacia than Eucalyptus, δ¹⁵N gave no clear indication for N₂ fixation in Acacia. In a multiple regression, latitude (as a surrogate for rainfall seasonality), mean rainfall, leaf nitrogen concentration, specific leaf area and nitrogen fixation were significant and explained 69 % of the variation of δ¹³C, but only 36 % of the variation of δ¹⁵N. Higher nitrogen and phosphorus concentration could give Acacia an advantage over Eucalyptus in arid regions of undefined rainfall seasonality.     

A new subspecies and new combinations in Comarostaphylis (Ericaceae)

Comarostaphylis spinulosa subsp.glandulifera is described from the mountains of Oaxaca and Puebla, Mexico. Four new combinations inComarostaphylis are also made:C. arbutoides subsp.costaricensis, C. discolor subsp.rupestris, C. polifolia subsp.minor, andC. spinulosa.Arctostaphylos glabrata, A. rupestris, andA. spinulosa are lectotypified and the status ofC. polifolia is discussed.     

Sequence and functional analyses of the aldehyde dehydrogenase 7B4 gene promoter in Arabidopsis thaliana and selected Brassicaceae: regulation patterns in response to wounding and osmotic stress

Key message

The core promoter of the antiquitin ALDH7B4 gene was compared between selected Brassicaceae. Conserved cis elements controlling osmotic stress and wound-induced expression were identified and analysed in Arabidopsis thaliana leaves and seeds.

Abstract

Aldehyde dehydrogenases metabolise a wide range of aliphatic and aromatic aldehydes, which become cytotoxic at high levels. Family 7 aldehyde dehydrogenase genes, often described as antiquitins or turgor-responsive genes in plants, are broadly conserved across all domains. Despite the high conservation of the plant ALDH7 proteins and their importance in stress responses, their regulation has not been investigated. Here, we compared ALDH7 genes of different Brassicaceae and found that, in contrast to the gene organisation and protein coding sequences, similarities in the promoter sequences were limited to the first few hundred nucleotides upstream of the translation start codon. The function of this region was studied by isolating the core promoter of the Arabidopsis thaliana ALDH7B4 gene, taken as model. The promoter was found to be responsive to wounding in addition to salt and dehydration stress. Cis-acting elements involved in stress responsiveness were analysed and two conserved ACGT-containing motifs proximal to the translation start codon were found to be essential for the responsiveness to osmotic stress in leaves and in seeds. The integrity of an upstream ACGT motif and a dehydration-responsive element/C-repeat—low temperature-responsive element was found to be necessary for ALDH7B4 expression in seeds and induction by salt, dehydration and ABA in leaves. The comparison of the gene expression in selected Arabidopsis mutants demonstrated that osmotic stress-induced ALDH7B4 expression in leaves and seeds involves both ABA- and lipid-signalling components.     

Rubiacearum Americanarum Magna Hama Pars XXXII. New Species and a New Combination in Coussarea (Coussareeae) from Western South America

The Neotropical genus Coussarea is species-rich and widely distributed but not well known. Seven new species are described here, documented mainly by recent botanical exploration: C. acrensis is found in western Brazil, C. boliviensis and C. mexiae in the mountains of northwestern Bolivia, and C. camposiana, C. maranonensis, C. pseudopilosula, and C. vasqueziana in Peru. The new combination C. insolita is based on Rudgea insolita from Colombia.