Two novel eukaryotic translation initiation factor 5A genes from <Emphasis Type="Italic">Populus simonii</Emphasis> × <Emphasis Type="Italic">P. nigra</Emphasis> confer tolerance to abiotic stresses in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

The role of plant eIF5A proteins in multiple biological processes, such as protein synthesis regulation, translation elongation, mRNA turnover, programmed cell death and stress tolerance is well known. Toward using these powerful proteins to increase stress tolerance in agricultural plants, in the present study, we cloned and characterized PsneIF5A2 and PsneIF5A4 from young poplar (P. simonii × P. nigra) leaves. The deduced amino acid sequences of PsneIF5A2 and PsneIF5A4 were 98 % similar to each other, and they are orthologs of eIF5A1 in Arabidopsis. In a subcellular localization analysis, PsneIF5A2 and PsneIF5A4 proteins were localized in the nucleus and cytoplasm. qRT-PCR analysis showed that PsneIF5A2 and PsneIF5A4 were transcribed in poplar flowers, stem, leaves, and roots. In addition, they were also induced by abiotic stresses. Transgenic yeast expressing PsneIF5A2 and PsneIF5A4 had increased salt, heavy metal, osmotic, oxidative tolerance. Our results suggest that PsneIF5A2 and PsneIF5A4 are excellent candidates for genetic engineering to improve salt and heavy metal tolerance in agricultural plants.     

A recommended rate for application of <Emphasis Type="Italic">Chaetomium globosum</Emphasis> ND35 fungus fertilizer on poplar plantations in China

Previous studies showed that Chaetomium globosum ND35 fungus fertilizer can improve the microbial community structure and enzyme activities of replanted soil. However, it remains unclear whether can improve the physiological and ecological characteristics of plants under successive rotation. In this study, we investigated the photosynthetic, physiological, and biochemical indexes including photosynthetic parameters, chlorophyll fluorescence, and chlorophyll content of 1-year-old poplar seedlings under seven different doses (range from 0 to 1.67 g kg^?1) of C. globosum ND35 fungus fertilizer to study the effects of fungus fertilizer on photosynthesis of Poplar. Our results showed that: (1) With increasing application of fungus fertilizer in replanted soil, chlorophyll content of poplar leaves (Chl) increased, while physiological indexes such as electron transport rate (ETR), net photosynthetic rate (P _n), quantum efficiency (Φ), nitrate reductase (NR) activity and root vigor initially increased and then declined. Meanwhile, heat dissipation that depended on the xanthophyll cycle declined and non-photochemical quenching (NPQ) initially increased and then decreased. When the dose of C. globosum ND35 fungus fertilizer was 0.67 g kg^?1 (T3) and 1.00 g kg^?1 (T4), excess light energy of photosynthetic apparatus was reduced, and photosynthetic apparatus distributed more light energy to the direction of photochemical reactions, which improved the efficiency of energy use. Plant height and biomass of leaves, stems, and roots were maximum at T3. We conclude that applying appropriate amounts of C. globosum ND35 fungus fertilizer can improve root physiological activity and capacity for use of light by poplar leaves. This can improve the operating states of the photosynthetic apparatus and lead to increased photosynthetic efficiency of poplar leaves and accumulation of dry matter. This suggests a strategy to alleviate the successive rotation obstacle of soil nutrient depletion.     

Biochemical changes associated with flowering in <Emphasis Type="Italic">Bambusa arundinacea</Emphasis> and <Emphasis Type="Italic">Bambusa nutans</Emphasis>

Biochemical changes associated with flowering in Bambusa arundinacea Linn and Bambusa nutans Wall. ex Munro were analyzed. Gregarious flowering was initiated in natural areas and plantations of B. arundinacea in late 2014 and reached full bloom in early 2015, whereas sporadic flowering recorded during 2014–2015 in vegetatively propagated plants of B. nutans. Leaf and nodal shoot samples from flowering culms/shoots and nonflowering culms/shoots were collected in February 2015 at the initiation of fruit filling, then analyzed and compared for soluble sugars, phenols and peroxidase activity. In both species sugars were higher in leaves of flowering culms/shoots and lower in nodal shoots of flowering culms/shoots compared to the nonflowering. Phenols were lower in leaves and nodal shoots of flowering B. arundinacea, but higher in leaves and decreases in nodal shoots of B. nutans. Peroxidase activity increases in leaves and nodal shoots of B. arundinacea after flowering but increases in nodal shoots and decreases in leaves of B. nutans.     

Identification and evaluation of reference genes for normalization in quantitative real-time PCR analysis in the premodel tree <Emphasis Type="Italic">Betula luminifera</Emphasis>

Betula luminifera is a commercial tree species that is emerging as a new model system for tree genomics research. A draft genomic sequence is expected to be publicly available in the near future, which means that an explosion of gene expression studies awaits. Thus, the work of selecting appropriate reference genes for qPCR normalization in different tissues or under various experimental conditions is extremely valuable. In this study, ten candidate genes were analyzed in B. luminifera subjected to different abiotic stresses and at various flowering stages. The expression stability of these genes was evaluated using three distinct algorithms implemented using geNorm, NormFinder and BestKeeper. The best-ranked reference genes varied across different sample sets, though RPL39, MDH and EF1a were determined as the most stable by the three programs among all tested samples. RPL39 and EF1a should be appropriate for normalization in N-starved roots, while the combination of RPL39 and MDH should be appropriate for N-starved stems and EF1a and MDH should be appropriate in N-starved leaves. In PEG-treated (osmotic) roots, MDH was the most suitable, whereas EF1a was suitable for PEG-treated stems and leaves. TUA was also stably expressed levels in PEG-treated plants. The combination of RPL39 and TUB should be appropriate for heat-stressed leaves and flowering stage. For reference gene validation, the expression levels of SOD and NFYA-3 were investigated. This work will be beneficial to future studies on gene expression under different abiotic stress conditions and flowering status in B. luminifera.     

Effects of flooding stress on the photosynthetic apparatus of leaves of two <Emphasis Type="Italic">Physocarpus</Emphasis> cultivars

We applied under pot-culture conditions and the double-casing pot method to study the characteristics of photosynthetic gas exchange and chlorophyll fluorescence in the leaves of Physocarpus amurensis Maxim (PA) and Physocarpus opulifolius under flooding stress. Our results indicate a significantly higher flooding tolerance of P. opulifolius compared to P. amurensis. Especially in P. amurensis, the limitation of non-stomatal factors played a major role in the advanced stages of flooding stress, observed as a rapid increase of the intercellular CO₂ concentration (C _i) and a decrease of the stomatal limitation value (L _s). The maximal PSII photochemical efficiencies (F _v/F _m) and actual photochemical efficiency (Ф _PSII) in the leaves of P. opulifolius were significantly higher, and the extent of decrease during the flooding process was smaller than in P. amurensis. In addition, the non-chemical quenching (NPQ) in the leaves of P. opulifolius significantly increased from the 10th day under flooding stress, while the variation of NPQ in the leaves of P. amurensis was much smaller. This indicates that the leaves of P. opulifolius had not only higher PSII photochemical activity, but also improved tolerance to flooding stress, which may be caused by its ability to dissipate excess excitation energy by starting NPQ. At the 16th day under flooding stress, the P _IABS significantly decreased with greater extent of decrease than F _v/F _m in the leaves of both Physocarpus, but the decreasing extent of P _IABS in P. opulifolius was significantly smaller than in P. amurensis. In the 16th day under flooding stress, the fluorescence at J and I point (V _J and V _I) in P. amurensis were significantly higher, and the extent of increase in V _J was greater than V _I. However, the variations of V _J and V _I in the leaves of P. opulifolius were smaller, suggesting that the damage sites of flooding stress to PSII in the leaves of P. amurensis were mainly located in the electron transport process from Q_A at the PSII receptor side to Q_B. Flooding stress reduced the proportion (φE _o ) of luminous energy absorbed by the PSII reaction center for the electron transport following Q _A ^? , while the maximum quantum yield (φD _o) of non-photochemical quenching increased. However, the TR_o/RC and ET_o/RC in the leaves of P. amurensis decreased accompanied by a dramatic increase of energy (DI_o/RC) from the dissipation in the reaction center. This further indicated that the function of the PSII reaction center in the leaves of P. amurensis was significantly lower than in P. opulifolius.     

The effects of phenolic acid on nitrogen metabolism in <Emphasis Type="Italic">Populus</Emphasis> × <Emphasis Type="Italic">euramericana</Emphasis> ‘Neva’

The declines in soil fertility and productivity in continuously cropped poplar plantations are related to phenolic acid accumulation in the soil. Nitrogen is a vital life element for poplar and whether the accumulation of phenolic acid could influence nitrogen metabolism in poplar and thereby hinder continuous cropping is not clear. In this study, poplar cuttings of Populus × euramericana ‘Neva’ were potted in vermiculite, and phenolic acids at three concentrations (0X, 0.5X and 1.0X) were added according to the actual content (1.0X) in the soil of a second-generation poplar plantation. Each treatment had eight replicates. We measured gas exchange parameters and the activities of key enzymes related to nitrogen metabolism in the leaves. Leaf photosynthetic parameters varied with the concentration of phenolic acids. The net photosynthetic rate (P_N) significantly decreased with increasing phenolic acid concentration, and non-stomatal factors might have been the primary limitation for P_N. The activities of nitrate reductase (NR), glutamine synthetase (GS) and glutamate synthase (GOGAT), as well as the contents of nitrate nitrogen, ammonium nitrogen, and total nitrogen in the leaves decreased with increasing phenolic acid concentration. This was significantly and positively related to P_N (P < 0.05). The low concentration of phenolic acids mainly affected the transformation process of NO₃^? to NO₂^?, while the high concentration of phenolic acids affected both processes, where NO₃^? was transferred to NO₂^? and NH₄⁺ was transferred to glutamine (Gln). Overall, phenolic acid had significant inhibitory effects on the photosynthetic productivity of Populus × euramericana ‘Neva’. This was probably due to its influence on the activities of nitrogen assimilation enzymes, which reduced the amount of amino acids that were translated into protein and enzymes. Improving the absorption and utilization of nitrogen by plants could help to overcome the problems caused by continuous cropping.     

An assessment of potential of hybrid poplar for planting in the Virginia Piedmont

Poplar species grow well across the temperate zone, but hybrid varieties have not previously been evaluated for planting in the Virginia Piedmont region. The top 12 clones in height growth and rust resistance from a screening trial involving 98 hybrid poplar varieties of three taxa (Populus deltoides × P. maximowiczii, DxM; P. deltoides × P. nigra, DxN; P. deltoides × P. trichocarpa, DxT) were selected for planting in replicated yield trials at two locations in the Virginia Piedmont. Results through the first four growing seasons showed that the DxM taxon had the most rapid height development. It was, however, the taxon most affected by a late spring frost at the Appomattox-Buckingham State Forest site and by Septoria stem canker at the Reynolds Homestead site. Analysis of variance of clonal and location effects showed highly significant differences among replicates within location and among clones within taxon. Among seven clones within the DxM taxon, pairwise comparison tests of height growth identified two groups: a group of four better clones that were significantly different (p = 0.05) from a second group of three. These early results suggest that multi-selection criteria, including growth, disease and frost resistance, are important when developing hybrid poplar clones for planting in the Piedmont region.     

<Emphasis Type="Italic">Orius laevigatus</Emphasis> strengthens its role as a biological control agent by inducing plant defenses

Orius laevigatus is a generalist predator that is widely used in augmentative strategies against the key pest of sweet pepper, Frankliniella occidentalis. Despite being a zoophytophagous predator, the phytophagous behavior of O. laevigatus has not been previously explored in depth nor has the impact of phytophagy on plant physiology. Here, the hierarchical significance of O. laevigatus feeding on sweet pepper is compared with other behaviors. O. laevigatus spends the majority of its time (38%) feeding on apical meristems and apical fresh leaves, which were also preferred residence locations. Here, the phytophagous feeding behavior of O. laevigatus on sweet pepper is shown to trigger defensive responses in the plant. These O. laevigatus plant-induced defenses are then shown to contribute to the repellence or attraction of pests or natural enemies, respectively. Specifically, O. laevigatus-punctured sweet pepper plants induce repellency for the whitefly Bemisia tabaci and the thrips species F. occidentalis. In contrast, the whitefly parasitoid Encarsia formosa was significantly attracted to O. laevigatus-punctured plants. The plant responses to O. laevigatus punctures include the release of an altered blend of volatiles and activation of the jasmonic acid and salicylic acid signalling pathways. These results highlight an interesting facet to the biology of O. laevigatus, in which the ability of the predator to induce defensive responses in sweet pepper plants may serve to improve the biological control of both thrips and whiteflies.     

Effects of <Emphasis Type="Italic">Verticillium nonalfalfae</Emphasis> on <Emphasis Type="Italic">Ailanthus altissima</Emphasis> and associated indigenous and invasive tree species in eastern Austria

The naturally occurring Verticillium nonalfalfae shows promise for biocontrol of the highly invasive Tree of Heaven (Ailanthus altissima), but might also bear a risk for non-target tree species. In this study, we conducted inoculations on potted seedlings of A. altissima as well as on eight indigenous and two invasive tree species associated with Tree of Heaven in Austria. Although vascular discolourations developed in all inoculated tree species, V. nonalfalfae was reisolated from Ailanthus and eight of the ten non-target-species, whereas typical disease symptoms and mortality only occurred on A. altissima. Results confirmed high susceptibility (S) of A. altissima to V. nonalfalfae but indicated tolerance (T) of Acer campestre, Acer pseudoplatanus and Quercus robur, possible resistance (PR) of Fraxinus excelsior, Populus nigra, Tilia cordata, Ulmus laevis and Ulmus minor and resistance (R) of Fraxinus pennsylvanica and Robinia pseudoacacia to this potential biocontrol agent. Results from seedling inoculations were confirmed by cursory field observations in Ailanthus-inoculated forest stands, where admixed A. campestre, A. pseudoplatanus, F. excelsior, Populus alba, R. pseudoacacia and U. laevis canopy trees remained asymptomatic, while mortality was induced in Ailanthus.     

The susceptibility of Asian,European and North American <Emphasis Type="Italic">Fraxinus</Emphasis> species to the ash dieback pathogen <Emphasis Type="Italic">Hymenoscyphus fraxineus</Emphasis> reflects their phylogenetic history

In Europe, common ash (Fraxinus excelsior) is being decimated because of the invasive fungus Hymenoscyphus fraxineus. In its native range in Asia this ascomycete is considered a harmless leaf associate of F. mandshurica and F. chinensis subsp. rhynchophylla. Field observations from Europe suggest that there is species-specific variation in disease susceptibility among European and North American Fraxinus species, but a wider comparison at the genus level has been missing so far. We assessed disease symptoms and pathogen apothecium development in 17 Fraxinus species from Asia, Europe and North America exposed to high infection pressure in a Danish arboretum. We also tested their susceptibility to pathogen infection through controlled stem and leaf inoculations and subsequently assessed the level of pathogen DNA by a qPCR assay. The results suggested the presence of a phylogenetic signal in disease susceptibility where closely related Asian, European and North American species in section Fraxinus had relatively high levels of H. fraxineus DNA in the leaves and supported high production of apothecia. Leaves from some North American species also contained relatively high levels of H. fraxineus DNA, supported moderate production of apothecia and developed lesions—stating the need to avoid introduction of H. fraxineus to North America.     

No consistent daily variation in DNA methylation detected in Populus nigra leaves by methylation-sensitive amplification polymorphism analysis

DNA methylation, an epigenetic mechanism used by cells to control gene expression, has an important biological role in plant development and environmental fitness. Since plant DNA methylation is closely related to environmental conditions, variation during the day is expected. Here, in genetically identical plants of Populus nigra clone N46, DNA methylation changes in leaves over a 24 h period were detected using the methylation-sensitive amplification polymorphism method. The results showed different DNA methylation patterns in mature poplar leaves: not only in individuals at the same time, but also in samples at each of the six time during the day. In addition,night samples had a higher percentage of methylation than in morning samples. However, no statistically significant differences were found among the samples gathered at different times. Similar results were obtained for three other P. nigra clones with different genetic backgrounds.Real time q PCR showed that the DNA methyltransferase genes Pt-MET1 and Pt-SOM1 involved in CG DNA methylation in poplar were stable over a 24 h period in leaves of P. nigra N46 compared with circadian-controlled genes. That could be part of the reason that methylation of CCGG sites is stable in those leaves. That DNA methylation differed even in genetically identical plants indicates the specificity of DNA methylation changes in their genomes. No statistically significant differences in methylation changes were found between day and night, suggesting that DNA methylation is more stable than expected and is unlikely to be involved in circadian regulation in plants.     

Within- and between-tree variation of wood density components in <Emphasis Type="Italic">Pinus nigra</Emphasis> at six sites in Portugal

Key message

In Europe, P. nigra wood presents a density pattern of longitudinal variation with an increase from east to west. However, no latitudinal tendencies were detected. Compared to other Portuguese resinous species, P. nigra revealed higher density, identical radial growth and intra-ring heterogeneity, which presents advantages for industry purposes. The environmental factors (Sites effect) manifest more strongly in the latewood components while the Trees/Sites effect is more strongly expressed in the earlywood components.

Context

Although P. nigra Arnold is one of the most important conifers in Europe, little is known about the wood’s characteristics in the southwest European region.

Aims

Our aims are to outline a first approach to study the growth and wood quality in P. nigra in Portugal comparing to other European natural stands and other resinous species.

Methods

Inter- and intra-wood density variation of P. nigra from six Portuguese sites was studied using microdensitometry. Analysis of variance (ANOVA) was performed in three subsets: 50 common rings, core (juvenile wood) and peripheral analysis (mature wood).

Results

The average ring density was 0.588 g cm^?3, with maximum values in the north and low altitudes. Regarding growth traits, no latitudinal and altitudinal tendencies were detected. Compared to the main timber species in Portugal (P. pinaster Aiton), P. nigra showed similar radial growth, higher density but lower intra-ring density homogeneity. The Sites effect mainly influenced latewood density components, while the Trees/Sites effect primarily influenced earlywood components. The Rings effect was found to be relatively low, with a density decrease in the tree’s first years followed by an increase in the periphery. Growth traits showed a reduction from pith to bark.

Conclusion

Considering the quality (density) and growth features of the Black pine, this species could be useful for the reforestation of mountainous Southern Europe areas that are not favourable for other species.

     

A novel cold-inducible promoter,P<Emphasis Type="Italic">ThCAP</Emphasis> from <Emphasis Type="Italic">Tamarix hispida</Emphasis>, confers cold tolerance in transgenic <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Our previous studies have revealed that the ThCAP gene plays a vital role in transgenic Populus (P. davidiana × P. bolleana) in response to cold stress. However, the regulatory mechanism of ThCAP gene expression has been unclear. In this study, the 5′ flanking region of the ThCAP promoter (PThCAP) was cloned using a genome-walking method. By analyzing cis-acting regulatory elements of PThCAP, a DRE motif and MYC and MYB elements were found to be located in the promoter. To identify the regulatory elements that control the expression of the ThCAP gene promoter, a series of deletion derivatives of PThCAP, P1–P5, from the translation start code (?1538, ?1190, ?900, ?718 and ?375 bp), were fused to the GUS reporter gene, and then each deletion was stably introduced into Arabidopsis thaliana plants. Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A. thaliana exposed to low temperature stress. These results suggest that this 290-bp region (?1190 to ?900 bp), as an important part in PThCAP, was associated with cold tolerance of A. thaliana. Our results provide evidence for the regulatory mechanism of ThCAP gene involved in the response to cold stress, and that the gene is promising candidate gene for genetic improvement of crops.     

Effects of extreme soil water stress on photosynthetic efficiency and water consumption characteristics of <Emphasis Type="Italic">Tamarix chinensis</Emphasis> in China’s Yellow River Delta

Soil moisture is a major limiting factor for plant growth on shell ridge islands in the Yellow River Delta. However, it is difficult to carry out situ experiment to study dominant plant photosynthesis physiological on the shell ridge islands under extreme soil water stress. To evaluate the adaptability of plants to light and moisture variations under extreme soil moisture conditions present on these islands, we measured photosynthetic gas exchange process, chlorophyll fluorescence, and stem sap flow variables for 3-year-old trees of Tamarix chinensis Lour, a restoration species on these islands, subjected to three types of soil water levels: waterlogging stress (WS), alternating dry–wet (WD), and severe drought stress (SS) to inform decisions on its planting and management on shell ridge islands. Gas exchange, chlorophyll fluorescence, and stem sap flow in T. chinensis were then measured. Net photosynthetic rate (P _N), transpiration rate (E), and water use efficiency (W _UE) were similar under WS and alternating dry–wet conditions, but their mean E and W _UE differed significantly (P < 0.05). Under SS, the P _N, E and W _UE of T. chinensis leaves varied slightly, and mean P _N, E and W _UE were all low. Apparent quantum efficiency (A _QY), light compensation point (L _CP), light saturation point (L _SP), and maximum net photosynthetic rate (P _Nmax) of leaves were not significantly different (P > 0.05) under WS and dry–wet conditions; however, under extreme drought stress, compared with the dry–wet conditions, L _CP was higher, L _SP was lower, and A _QY and P _Nmax were both at the lowest level. Therefore, drought stress weakened light adaptability of leaves, and the efficiency of light transformation was poorer. (3) Maximum photochemical efficiency (F _v/F _m) and the actual photochemical efficiency (Φ _PSII) were similar under waterlogged stress and dry–wet conditions, indicating a similar healthy photosynthetic apparatus and photosynthetic reaction center activity, respectively. Under SS, F _v/F _m was 0.631, and the coefficient of non-photochemical quenching (N _PQ) was 0.814, which indicated that while the photosynthetic mechanism was damaged, the absorbed light energy was mainly dissipated in the form of heat, and the potential photosynthetic productivity was significantly reduced. The daily cumulants of sap flow of T. chinensis under dry–wet alternation and severe drought stress were 22.25 and 63.97% higher, respectively, than under waterlogging stress. Daily changes in sap flow velocity for T. chinensis differed under the three soil water levels. Stem sap flow was weak at night under severe drought stress. Under dry–wet alternation, daytime average stem sap flow velocity was the highest, and night stem flow accounted for 10.26% of the day cumulants, while under waterlogged stress, the average nightly stem flow velocity was the highest, accounting for 31.82% of the day cumulants. These results provide important information for regional vegetation restoration and ecological reconstruction.     

Genetic diversity and population structure analysis of <Emphasis Type="Italic">Pistacia</Emphasis> species revealed by <Emphasis Type="Italic">phenylalanine ammonia</Emphasis>-<Emphasis Type="Italic">lyase</Emphasis> gene markers and implications for conservation

Information on population genetic structure and crop genetic diversity is important for genetically improving crop species and conserving threatened species. The PAL gene sequence is part of a multigene family that can be utilized to design DNA marker systems for genetic diversity and population structure investigation. In the current study, genetic diversity and population structure of 100 accessions of wild Pistacia species were investigated with 78 PAL markers. A protocol for using PAL sequences as DNA markers was developed. A total of 313 PAL loci were recognized, showing 100% polymorphism for PAL markers. The PAL markers produced relatively more observed and effective alleles in Pistacia falcata and Pistacia atlantica, with a higher Shannon’s information index and expected heterozygosity in P. atlantica, Pistacia vera and Pistacia mutica. Pairwise assessment of Nei’s genetic distance and genetic identity between populations revealed a close association between geographically isolated populations of Pistacia khinjuk and Pistacia chinensis. The accessions of wild Pistacia species had more genetic relationship among studied groups of species. Analysis of molecular variance indicated 19% among-population variation and 81% within-population variation for the PAL gene based DNA marker. Population structure analysis based on PAL revealed four groups with high genetic admixture among populations. The results establish PAL markers as a functional DNA marker system and provide important genetic information about accessions from wild populations of Pistacia species.     

Application of oxygen-sensing technology to measure seed quality of <Emphasis Type="Italic">Ginkgo biloba</Emphasis>

Germination tests are currently the most widely used method to evaluate seed quality of Ginkgo biloba L., but they are time-consuming and labor intensive. Oxygen-sensing technology, based on the principle of fluorescence quenching to detect oxygen and assess seed quality was used to rapidly evaluate seed quality of two varieties (Shandong Tancheng 202 and Zhejiang Dafoshou) of G. biloba from five mother plants. Fifteen samples of three vigor levels were produced by accelerated aging treatments. This process was applied in duplicate. A portable oxygen-sensing detector was employed to measure oxygen content during seed germination in a closed system at 25 °C each day until day 14. Four oxygen metabolism indices were calculated: oxygen consumption index, oxygen consumption rate, critical oxygen concentration, and theoretical germination time (T _GT). Additionally, we tested laboratory germination and field emergence. The results of a one-way analysis of variance and correlation analysis showed that T _GT was the candidate index to evaluate seed quality of G. biloba. Therefore, the T _GT value was used to validate the reliability of oxygen-sensing evaluation for Zhejiang Dafoshou seeds kept under four storage conditions. The trend in the change in oxygen metabolism agreed completely agreement with that of seed germination under all storage conditions. The oxygen-sensing test reliably and rapidly assessed seed quality of G. biloba. The germination rate of Zhejiang Dafoshou was accurately predicted by T _GT.     

<Emphasis Type="Italic">Vicia faba</Emphasis> plants respond to oviposition by invasive <Emphasis Type="Italic">Halyomorpha halys</Emphasis> activating direct defences against offspring

The invasive stink bug Halyomorpha halys is established in many European and American agro-ecosystems, where it causes severe crop losses. Potential control measures might include enhancement of plant defences. When attacked by herbivorous insects that oviposit on it, the plant may respond by priming direct defences, which might affect the development of future brood. Halyomorpha halys attacks numerous plant species in the invaded areas. Here, we investigated whether Vicia faba plants challenged by H. halys females can impair the development of its offspring through the activation of induced direct defences. We measured the weight and dimension of nymphs that developed on oviposition-experienced plants after 7 and 17 days from hatching. Nymphs that developed on oviposition-experienced plants weighed less compared to those that developed either on control plants or on plants solely subjected to H. halys feeding, and third instars showed shorter dimensions (tibia length). In addition, when oviposition-experienced plants were attacked by nymphs, higher and more rapid expression of two jasmonic acid-dependent genes (cysteine proteinase inhibitor gene and NAI1) was detected, possibly due to a mechanism of priming. Increased expression of the salicylic acid-responsive PR1 gene was also detected in egg-experienced plants, although the response was delayed compared to JA-dependent genes. Our results suggest that V. faba plants recognize H. halys oviposition as a warning signal and pre-activate defences against future nymphal herbivory.