A linkage map of maize × teosinte <Emphasis Type="Italic">Zea luxurians</Emphasis> and identification of QTLs controlling root aerenchyma formation

The objectives of this study were to construct a linkage map and identify quantitative trait loci (QTLs) controlling root aerenchyma formation in drained soil conditions using 195 F₂ individuals derived from a cross between maize inbred line B73 × teosinte Zea luxurians. A 107 SSR marker based map covering 1,331 cM across all ten chromosomes was developed. One significant difference between the parents utilized in the study was that under non-flooding conditions, B73 exhibits a minor capacity to develop root aerenchyma, whereas Z. luxurians exhibits a high tendency to form aerenchyma. Linkage analysis indicated segregation distortion regions on chromosomes 2, 4 and 8, and severe recombination suppression on the long arm of chromosome 4. Multiple interval mapping analysis suggests that five QTLs for aerenchyma formation in non-flooding conditions are located on chromosomes 2, 3, 5, 9 and 10, and these explained 36.3% of the total phenotyphic variance. The Z. luxurians alleles in all five QTLs increased the capacity to form aerenchyma and the locations of these QTLs did not overlap those previously identified in the teosinte Z. nicaraguensis. By transferring aerenchyma-forming QTLs from both Z. luxurians and Z. nicaraguensis, it may be possible to pyramid these genes and develop a maize line with exceptional aerenchyma formation and a high level of tolerance to flooding conditions.     

A diazotrophic bacterial endophyte isolated from stems of Zea mays L. and Zea luxurians Iltis and Doebley

The apoplastic fluids of field-grown Zea mays and Zea luxurians plants were isolated from surface sterilized stem tissue by centrifugation and spread on agar plates containing a nitrogen-free, defined medium. The predominant bacterium isolated from these plates was characterized further. The ability of this bacterium to fix nitrogen was confirmed by its ability to grow on a semi-solid, nitrogen-free medium and reduce ¹⁵N₂ to ¹⁵NH₃ and acetylene to ethylene. Protions of the nifH and 16S rRNA genes from this organism were amplified by PCR and sequenced. The nifH gene, which codes for dinitrogenase reductase, from this organism is closely related to nifH from Klebsiella pneumoniae. Similarly, the 16S rRNA gene sequences and carbon utilization tests grouped it closely with K. pneumoniae. Based an these data, the isolates from Z. mays and Z. luxurians are tentatively classified as Klebsiella spp. (Zea). The ability of this bacterium to contribute to the nitrogen economy of the corn plant is unknown.     

Variation within teosinte. III. numerical analysis of allozyme data

Principal components analysis of isozyme allele frequencies at 19 loci revealed 133 electrophoretic variants for 77 accessions of annual teosinte and 1 accession each of diploid and tetraploid perennial teosintes. The majority of alleles were found in low frequency, and many were distributed only in specific locations. Zea luxuriansand the annual Mexican teosintes appeared to be the most distantly related of all teosintes. Z. perennisand Z. diploperennisappeared distinct from each other and from all other teosintes. Teosintes of west Guatemala (Huehuetenango) did not appear especially closely related to Balsas teosintes. Several differences were apparent between Chalco and Central Plateau teosintes;however, these differences were not so extreme as those suggested by chromosome knob data. Nobogame teosintes appeared closely related to Chalco and Central Plateau teosintes. Isozyme data reveal teosinte to be a diverse source of germplasm.     

The NLRomes of Zea mays NAM founder lines and Zea luxurians display presence–absence variation,integrated domain diversity,and mobility

Plant pathogens cause significant crop loss worldwide, and new resistance genes deployed to combat diseases can be overcome quickly. Understanding the existing resistance gene diversity within the germplasm of major crops, such as maize, is crucial for the development of new disease-resistant varieties. We analysed the nucleotide-binding leucine-rich repeat receptors (NLRs) of 26 recently sequenced diverse founder lines from the maize nested association mapping (NAM) population and compared them to the R gene complement present in a wild relative of maize, Zea luxurians. We found that NLRs in both species contain a large diversity of atypical integrated domains, including many domains that have not previously been found in the NLRs of other species. Additionally, the single Z. luxurians genome was found to have greater integrated atypical domain diversity than all 26 NAM founder lines combined, indicating that this species may represent a rich source of novel resistance genes. NLRs were also found to have very high sequence diversity and presence–absence variation among the NAM founder lines, with a large NLR cluster on Chr10 representing a diversity hotspot. Additionally, NLRs were shown to be mobile within maize genomes, with several putative interchromosomal translocations identified.     

A Tea Hydroperoxide Lyase Gene,CsiHPL1, Regulates Tomato Defense Response Against Prodenia Litura (Fabricius) and Alternaria Alternata f. sp. Lycopersici by Modulating Green Leaf Volatiles (GLVs) Release and Jasmonic Acid (JA) Gene Expression

Hydroperoxide lyases (HPLs) play important roles in modulating plant defense by regulating the release of green leaf volatiles (GLVs) and the jasmonic acid (JA) pathway. CsiHPL1—a chloroplast-localized tea gene that encodes HPL—was previously cloned and predicted to be a regulator of plant defense responses. CsiHPL1 was expressed constitutively in transgenic tomato (Solanum lycopersicum) plants to define its function in plant defense. CsiHPL1 overexpression caused tomato to release more constitutive and wound-induced GLVs [including (Z)-hexenal and (Z)-3-hexen-1-ol]. CsiHPL1 transgenic lines also exhibited lower levels of resistance to the larva of the tomato-chewing herbivore Prodenia litura (Fabricius) but enhanced resistance to the necrotrophic fungus Alternaria alternata f. sp. lycopersici (AAL). Furthermore, transgenic lines exhibited decreased expression levels of JA-related genes (SlAOS and SlPI-II) induced by P. litura and AAL infection. We thus concluded that constitutive expression of CsiHPL1 can regulate tomato resistance to P. litura and AAL by modulating GLV release and JA gene expression. Application of these results will be helpful in controlling plant defenses against herbivore attack and fungal disease.     

Isolation and identification of TL-DNA/plant junctions in Convolvulus arvensis transformed by Agrobacterium rhizogenes strain A4

We have constructed a Charon 4A phage library containing insert DNA isolated from a morning glory (Convolvulus arvensis) plant (clone 7) regenerated from a root organ culture incited by Agrobacterium rhizogenes, strain A4. Using a subcloned region of the Ri plasmid as ³²P-labeled probe, two lambda clones containing most of the 'left' T-DNA (TL) region were isolated. One of these lambda clones contains the left TL-DNA/plant junction, which was located by comparing nucleotide sequences from the appropriate regions of the Ri plasmid and this lambda clone. A 25-bp sequence found near this left TL-DNA/plant junction matches the 25-bp terminal sequence found at or near T-DNA/plant junctions of both nopaline- and octopine-type A. tumefaciens Ti plasmids. A possible location for the right Ri TL-DNA/plant junction in C. arvensis clone 7 was found by obtaining the nucleotide sequence surrounding its mapped location. Hybridization of plant DNA found adjacent to the left TL-DNA/plant junction against total C. arvensis DNA shows that this T-DNA integration occurred in a plant DNA region that does not contain highly repetitive DNA sequences. Nucleotide sequence analysis of 1004 bp of this plant DNA revealed no complete or partial open reading frames, but this plant DNA does have the potential to form various secondary structures which might play a role in the T-DNA integration event.     

Plant N-glycan profiling of minute amounts of material

Development of convenient strategies for identification of plant N-glycan profiles has been driven by the emergence of plants as an expression system for therapeutic proteins. In this article, we reinvestigated qualitative and quantitative aspects of plant N-glycan profiling. The extraction of plant proteins through a phenol/ammonium acetate procedure followed by deglycosylation with peptide N-glycosidase A (PNGase A) and coupling to 2-aminobenzamide provides an oligosaccharide preparation containing reduced amounts of contaminants from plant cell wall polysaccharides. Such a preparation was also suitable for accurate qualitative and quantitative evaluation of the N-glycan content by mass spectrometry. Combining these approaches allows the profiling to be carried out from as low as 500 mg of fresh leaf material. We also demonstrated that collision-induced dissociation (CID) mass spectrometry in negative mode of N-glycans harboring α(1,3)- or α(1,6)-fucose residue on the proximal GlcNAc leads to specific fragmentation patterns, thereby allowing the discrimination of plant N-glycans from those arising from mammalian contamination.     

NADP-malate dehydrogenase from Zea mays leaves: Amino acid composition and thiol content of active and inactive forms

NADP-malate dehydrogenase (L-malate: NADP⁺ oxidoreductase, E.C. 1.1.1.82) was purified from the leaves of Zea mays L. and its subunit molecular weight, amino acid composition and the changes in number of thiol groups during activation were determined. The amino acid composition we found differed from that reported earlier for the Z. mays enzyme but was very similar to that reported for the enzyme isolated from pea leaves. The maize enzyme contains fewer methionine residues (3 compared to 5 in pea) but a greater total number of cysteine residues (6 compared to 3 in pea). In its inactive form (oxidised) the enzyme contained 2 thiols per subunit of which only 1 reacts with 5,5′-dithiobis(2-nitrobenzoic acid) when the enzyme is in its native form. During activation by dithiothreitol two disulphide bonds are reduced per subunit to give 4 new thiol groups. We conclude that NADP-malate dehydrogenase from leaves of the C₄ plant Z. mays is very similar to the enzyme from the C₃ plant pea. However, apparently two disulphide bonds are reduced during the reductive activation of the Z. mays enzyme in vitro compared with 1 disulphide bond for the pea enzyme.     

TAXONOMY OF ZEA (GRAMINEAE). I. A SUBGENERIC CLASSIFICATION WITH KEY TO TAXA

The genus Zea is here divided into the Sect. Luxuriantes Doebley & litis sect. n., including the perennials Z. diploperennis (2n = 20) and Z. perennis (2n = 40) and the annual Z. luxurians (2n = 20); and Sect. Zea , including the wild Z. mays ssp.parviglumis and Z. mays ssp. mexicana (both 2n = 20), and Z. mays ssp. mays (2n = 20), the highly domesticated and tremendously variable derivate of the latter. This division is verified by a multivariate analysis of a large number of morphological characters of the male inflorescence. Cytogenetic and chemotaxonomic evidence supports the morphological conclusions. A consideration of the phylogeny of Zea within the conceptual framework offered by this new sectioning of the genus points convincingly to annual teosinte (Z. mays ssp. mexicana) as the ancestor of cultivated maize.     

Repellent,larvicidal and adulticidal activities of essential oil from Dai medicinal plant Zingiber cassumunar against Aedes albopictus

Zingiber cassumunar is an important plant used in traditional medicine and as a natural mosquito repellent. However, the compounds responsible for the repellent activity of the plant are still unknown. The aim of the study is to identify the components of Z. cassumunar essential oil that show repellent activity against Aedes albopictus. We also evaluated the larvicidal and adulticidal activities of Z. cassumunar essential oil against Ae. albopictus. In-cage mosquito repellent experiments showed that Z. cassumunar essential oil possessed moderate repellent activity with a minimum effective dose (MED) of 0.16 ± 0.01 mg/cm², compared to reference standard N,N-diethyl-3-methylbenzamide (DEET, 0.03 ± 0.01 mg/cm²). Bioassay-guided fractionation identified the major active compound of Z. cassumunar essential oil as (−)-terpinen-4-ol (1) (MED: 0.19 ± 0 mg/cm²). We also found that Z. cassumunar essential oil showed moderate larvicidal activity against first instar larvae of Ae. albopictus with a LC₅₀ (50% lethal concentration) of 44.9 μg/L after 24 h. Fumigation bioassays showed that Z. cassumunar essential oil exhibits moderate adulticidal activity against Ae. albopictus with a LC₅₀ of 5.44%, while (−)-terpinen-4-ol showed significant adulticidal activity with a LC₅₀ of 2.10% after 24 h. This study verifies that the Z. cassumunar essential oil has mosquito repellent activity, and that (−)-terpinen-4-ol is mainly responsible for this activity. Furthermore, this study provides scientific support for the folk usage of Z. cassumunar essential oil as mosquito repellent and indicates that Z. cassumunar essential oil and (−)-terpinen-4-ol can be used as plant-derived repellents and insecticides for mosquito control.     

Leaf flavonoid and other phenolic glycosides as indicators of parentage in six ornamental Fuchsia species and their hybrids

In a leaf flavonoid analysis of six Fuchsia species and seven Fuchsia hybrids, flavonols were found to be abundant in all taxa except F. procumbens. Flavone glycosides were found in only three species: luteolin 7-glucoside in F. splendens; and luteolin and apigenin 7-glucuronides and 7-glucuronidesulphates, tricin 7-glucuronidesulphate and diosmetin 7-glucuronide from one or both of the New Zealand species F. procumbens and F. excorticata. Luteolin 7- glucuronidesulphate is reported for the first time. Other less common phenolics identified include the flavanone, eriodictyol 7-glucoside from F. excorticata, a galloylglucose from F. triphylla, and a galloylglucosesulphate present in all taxa. Eight of the flavonoid glycosides proved useful as marker substances for particular Fuchsia species: quercetin 3- rhamnoside, 3-glucuronide and 3-rutinoside for F.fulgens; quercetin and kaempferol 3-galactosides for F. boliviana var. luxurians; diosmetin 7-glucuronide for F. excorticata and apigenin 7-glucuronide and 7-glucuronidesulphate for F. procumbens. The chemical results on the hybrids support the view that the cultivar ‘Mary’ is a hybrid of F. boliviana var. luxurians and F. triphylla and that both F.fulgens and F. triphylla are involved as parents of the cultivars ‘Koralle’ and ‘Traudchen Bondstedt’.     

The use of medium point density LiDAR elevation data to determine plant community types in Baltic coastal wetlands

This study investigates the use of medium point density LiDAR (light detection and ranging) elevation data for identifying and locating plant community types in Baltic coastal wetlands. Ground-surveyed micro-topography data obtained using real-time kinematic differential GPS were used to analyse and correct the accuracy of LiDAR sourced elevation values in Matlab R2010a. Following the application of elevation corrections, 3 interpolation techniques were applied: Inverse Distance Weighting (IDW), Ordinary Kriging (OK) and the Delaunay Triangulated Irregular Network (TIN) interpolation. Analysis showed that the TIN method produced a more robust model that adequately represented the micro-topographical variation across the wetland. The TIN elevation interpolation was then used to characterise the extent and distribution of the plant communities using known elevation preferences in one coastal wetland site, Tahu. The plant community model developed from the TIN elevation interpolation was then further ground-truthed in two separate Baltic coastal wetland sites, Matsalu and Kudani, in order to test model validity.The results showed that the accuracy of the LiDAR data varied dependent upon plant community type. In 5 of the 6 studied community types, the LiDAR data gave a mean elevation difference of 0.177 m (0.016 SD) above the dGPS elevations. With dGPS elevation corrections, the LiDAR data produced an accurate model of the location of the different plant community types, Kappa coefficient (κ) 0.63, at the initial site, Tahu. The model was found to be very accurate in one of the subsequent ground-truthed sites, κ 0.81, with a similar management history, but less accurate, κ 0.53, in a wetland more exposed to wave action and with a longer history of vegetation management.The inclusion of dGPS ground survey data was shown to considerably improve the accuracy of the LiDAR elevation data to facilitate the identification of plant community types on the basis of elevation. The approach developed in this study of Baltic coastal wetlands may provide a transferable method for rapidly and accurately mapping plant communities in other open environments.     

Growth Patterns Inferred from Anatomical Records : Empirical Tests Using Longisections of Roots of Zea mays L

Our objective was to test whether accurate growth analyses can be obtained from anatomical records and some mathematical formulas. Roots of Zea mays L. were grown at one of two temperatures (19°C or 29°C) and were prepared with standard techniques for light microscopy. Positions of cell walls were digitized from micrographs. The digitized data were averaged and smoothed and used in formulas to estimate growth trajectories, Z(t), velocities, v(z), and strain rates, r(z), where Z(t) is the location occupied by the cellular particle at time t; and v(z) and r(z) are, respectively, the fields of growth velocity and strain rate. The relationships tested are: for Z(t), t = n * c; v(z) = l(z) * f; and r(z) = f * (/z (l(z))). In the formulas, n represents the number of cells between the origin and the position Z(t); l(z) is local cell length; the constant c, named the `cellochron,' denotes the time for successive cells to pass a spatial point distal to the meristem; l(z) is local cell length, and f is cell flux. Growth trajectories and velocity fields from the anatomical method are in good agreement with earlier analyses based on marking experiments at the two different temperatures. Growth strain rate fields show an unexpected oscillation which may be due to numerical artifacts or to a real oscillation in cell production rate.     

Mate-Searching Behaviour of Common and Rare Wasps and the Implications for Pollen Movement of the Sexually Deceptive Orchids They Pollinate

Pollinator behaviour directly affects patterns of pollen movement and outcrossing rates in plants. In orchids pollinated by sexual deception of insects, patterns of pollen movement are primarily determined by the mate-searching behaviour of the deceived males. Here, using a capture-mark-recapture study (CMR) and dietary analysis, we compare mate-searching behaviour in relation to local abundance of two pollinator species and explore the implications for pollen movement in sexually deceptive Drakaea (Orchidaceae). Drakaea are pollinated solely by the sexual deception of male thynnine wasps. The rare Drakaea elastica and widespread D. livida occur sympatrically and are pollinated by the rare but locally common Zaspilothynnus gilesi, and the widespread and abundant Z. nigripes, respectively. Local abundance was significantly different with Z. nigripes twice as abundant as Z. gilesi. For the 653 marked wasps, there was no significant difference in median movement distance between Z. gilesi and Z. nigripes. However, the maximum movement distance was twice as high for Z. gilesi (556 m) compared with Z. nigripes (267 m). This is up to three times greater than previously reported for thynnines in CMR studies. Recapture rates were six times higher in Z. gilesi (57%) compared to Z. nigripes (9%). Pollen loads and wasp longevity were similar, suggesting that this difference in recapture rate arises due to differences in the number of males moving at a scale >500 m rather than through diet or mortality. Differences in the frequency of longer movements may arise due to variation in the spatial distribution of the wingless females. We predict that pollen movement will largely be restricted to within populations of Drakaea (<500 m), with few movements between populations (>500 m).     

Moth sex pheromone receptors and deceitful parapheromones

The insect''s olfactory system is so selective that male moths, for example, can discriminate female-produced sex pheromones from compounds with minimal structural modifications. Yet, there is an exception for this “lock-and-key” tight selectivity. Formate analogs can be used as replacement for less chemically stable, long-chain aldehyde pheromones, because male moths respond physiologically and behaviorally to these parapheromones. However, it remained hitherto unknown how formate analogs interact with aldehyde-sensitive odorant receptors (ORs). Neuronal responses to semiochemicals were investigated with single sensillum recordings. Odorant receptors (ORs) were cloned using degenerate primers, and tested with the Xenopus oocyte expression system. Quality, relative quantity, and purity of samples were evaluated by gas chromatography and gas chromatography-mass spectrometry. We identified olfactory receptor neurons (ORNs) housed in trichoid sensilla on the antennae of male navel orangeworm that responded equally to the main constituent of the sex pheromone, (11Z,13Z)-hexadecadienal (Z11Z13-16Ald), and its formate analog, (9Z,11Z)-tetradecen-1-yl formate (Z9Z11-14OFor). We cloned an odorant receptor co-receptor (Orco) and aldehyde-sensitive ORs from the navel orangeworm, one of which (AtraOR1) was expressed specifically in male antennae. AtraOR1•AtraOrco-expressing oocytes responded mainly to Z11Z13-16Ald, with moderate sensitivity to another component of the sex pheromone, (11Z,13Z)-hexadecadien-1-ol. Surprisingly, this receptor was more sensitive to the related formate than to the natural sex pheromone. A pheromone receptor from Heliothis virescens, HR13 ( = HvirOR13) showed a similar profile, with stronger responses elicited by a formate analog than to the natural sex pheromone, (11Z)-hexadecenal thus suggesting this might be a common feature of moth pheromone receptors.     

Fluorescence imaging application: effect of leaf age on seagrass photokinetics

We used the Imaging-PAM fluorometer to map spatial variability of photosynthesis in three seagrass species, Halophila ovalis, Zostera capricorni and Posidonia australis. Photosynthesis was described by relative photosynthetic rate (PS/50), effective quantum yield (Φ_PSII), non-photochemical quenching (NPQ and qN), electron transport rate (ETR) and leaf absorptivity. Photosynthetic patterns were linked to leaf age and light climate but patterns were not consistent across species. Longitudinal heterogeneity in photosynthesis was apparent along the leaves of all three species while lateral spatial heterogeneity was found only across Z. capricorni and H. ovalis leaves. Age of leaf tissue, determined by longitudinal location on the leaf, strongly influenced photosynthetic activity of Z. capricorni and P. australis. A comparison of H. ovalis leaves of differing maturity demonstrated the influence of leaf age on photosynthetic activity, yet a comparison of Z. capricorni leaves of differing maturity showed no leaf-age effects.Variations in stress-induced changes across a seagrass leaf can be used to identify areas or particular regions of the leaf, which are more susceptible to photodamage. Clear evidence of substantial within-leaf heterogeneity in photosynthetic activity (i.e., a two-fold variation in half saturation constant along a leaf of P. australis) has serious implications for use of small sections of leaf for photosynthetic incubations (such as O₂ or single-point chlorophyll a fluorescence measurements).     

Evaluation of diffusion and dilution methods to determine the antibacterial activity of plant extracts

The aim of this study was to evaluate diffusion and dilution methods for determining the antibacterial activity of plant extracts and their mixtures. Several methods for measurement of the minimal inhibitory concentration (MIC) of a plant extract are available, but there is no standard procedure as there is for antibiotics. We tested different plant extracts, their mixtures and phenolic acids on selected gram-positive (Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes) and gram-negative bacteria (Escherichia coli O157:H7, Salmonella Infantis, Campylobacter jejuni, Campylobacter coli) with the disk diffusion, agar dilution, broth microdilution and macrodilution methods. The disk diffusion method was appropriate only as a preliminary screening test prior to quantitative MIC determination with dilution methods. A comparison of the results for MIC obtained by agar dilution and broth microdilution was possible only for gram-positive bacteria, and indicated the latter as the most accurate way of assessing the antimicrobial effect. The microdilution method with TTC (2,3,5-triphenyl tetrazolium chloride) or INT (2-p-iodophenyl-3-p-nitrophenyl-5-phenyl tetrazolium chloride) to indicate the viability of aerobic bacteria was found to be the best alternative approach, while only ATP determination was appropriate for microaerophilic Campylobacter spp. Using survival curves the kinetics of bacterial inactivation on plant extract exposure was followed for 24 h and in this way the MIC values determined by the microdilution method were confirmed as the concentrations of extracts that inhibited bacterial growth. We suggest evaluation of the antibacterial activity of plant extracts using the broth microdilution method as a fast screening method for MIC determination and the macrodilution method at selected MIC values to confirm bacterial inactivation. Campylobacter spp. showed a similar sensitivity to plant extracts as the tested gram-positive bacteria, but S. Infantis and E. coli O157:H7 were more resistant.     

Z-3-Hexenylacetate emissions induced by the endophyte Epichloë occultans at different levels of defoliation during the host plant's life cycle

Vertically transmitted fungal endophytes are common defensive symbionts of cool-season grasses. Protection against herbivores has been generally associated with alkaloids produced in the grass-endophyte symbiosis. However, many other changes occur in host metabolism like the release of VOCs. We aimed at characterizing the profile of volatile organic compounds (VOCs) induced by simultaneous fungal endophyte symbiosis and defoliation during the entire life cycle of the annual host grass and the asexual symbiont. We designed an outdoor factorial experiment with plots dominated by intact and damaged Lolium multiflorum plants with high and low infection levels with Epichloë occultans. After exploring the entire VOC profile, the green leaf volatile (Z)-3-hexenyl acetate (Z3-HAC) was found as the main compound emitted by the plants under field conditions. While in low-infected patches there were no differences in volatile emission during the plant life cycle, highly infected patches emitted more Z3-HAC in seedling and vegetative phases than in the reproductive phase. The role of Z3-HAC provided by symbiotic to neighbouring non-symbiotic plants in the associational protection against herbivores and fungal pathogens is discussed.