Hydrothermal time analysis of watermelon (<Emphasis Type="Italic">Citrullus vulgaris</Emphasis> cv. ‘Crimson sweet’) seed germination

This study evaluated the ability of a hydrothermal time model (HTT) to describe the kinetics of watermelon (Citrullus vulgaris cv. ‘Crimson sweet’) seed germination under different temperatures (T) and water potentials (ψ) and also to determine the cardinal temperatures of watermelon. Results indicated that ψ influenced germination rate and germination percentage. For this seed lot, cardinal temperatures were 10 °C for T _b, 28.34 °C for T _o and 40.8 °C for T _c in the control (0 MPa) treatment. There was a decrease in hydrotime constant (θ _H) when T was increased to T _o and then remained constant at supra-optimal temperatures (30 MPah^?1). Also, at temperatures above T _o, ψ _b(50) values increased linearly with T. The k _T value (the slope of the relationship between ψ _b(50) and T exceeds T _o) of this seed lot was calculated as 0.076 MPa°Ch^?1. Results this study show that when the HTT model is applied, it can accurately describe ψ _b(g) and the course of germination around Ts (R ² = 0.82). Moreover, the ψ _b(50) was estimated to be ?0.96 MPa based on this model. Consequently, the germination response of watermelon for all Ts and ψs can be adequately described by the HTT model and enabling it to be used as a predictive tool in watermelon seed germination simulation models.     

Quantification of soybean seed germination response to seed deterioration under PEG-induced water stress using hydrotime concept

This experiment was arranged to investigate the ability of hydrotime model (θ_H) for estimating soybean seed germination (cv. ‘JK’) under different accelerated aging periods (AAP, 0, 24, 48, and 72 h) at each of the following water potentials (ψ, 0, ??0.12, ??0.24, and ??0.36 MPa). Results indicated that both germination percentage (GP) and germination rate (GR) significantly influenced by ψ, AAP, and their interactions (P?<?0.01). GP and GR decreased by 62.6 and 47.3% with longer AAP from 0 to 72 h and by 90.7 and 81.5% with lower ψ from zero to ??0.36 MPa as compared to the control, respectively. Therefore, the effect of ψ on GP and GR was more than AAP. The θ_H value was constant (~?6.71 MPa h^?1) till 50.6 h AAP and then linearly declined with the rate of 0.1545 MPa h^?1 per hour increase in AAP until 72 h (~?50% lower than its initial value). The ψ_b(50) value was ? 0.343 MPa in the control and then increased (became more positive) by ~?70% until 72 h AAP (? 0.104 MPa). In general, GP and GR of soybean declined with the increasing ψ_b(50) which can be due to cell membrane damage and reduce the activity of enzymes and organelles during AAP. Based on our findings, the θ_H model could describe well these relationships and their parameters can nicely be used for simulating soybean seed germination under this condition.     

Using hydrothermal time concept to describe sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) seed germination response to temperature and water potential

To quantify both temperature (T) and water potential (ψ) effects on sesame (Sesamum indicum L.) seed germination (SG) and also to determine the cardinal T _s for this plant, a laboratory experiment was carried out using hydrothermal time model (HTT). For this purpose, four sesame cultivars (‘Asbomahalleh’, ‘Darab’, ‘Dashtestan’ and ‘Yellowhite’) were germinated at seven constant T _s (20, 25, 30, 35, 37, 39 and 43 °C) at each of the following ψ _s (0, ? 0.12, ? 0.24 and ? 0.36 MPa; provided by PEG 8000). Germination rate (GR) and germination percentage (GP) significantly influenced by ψ, T and their interactions in all cultivars (P ≤ 0.01). There was no significant difference, based on the confidence intervals of the model coefficients, between cultivars, so an average of cardinal T _s was 14.7, 35.4 and 47.2 °C for the minimum (T _b), optimum (T _o) and maximum (T _c) T _s, respectively, in the control condition (0 MPa). Hydrotime values in all cultivars decreased when T was increased to T _o and then remained constant at T _s > T _o (15 MPa h^?1). An average value of ψ _b(50) was estimated to be ? 1.23 MPa at T _s ≤ T _o and then increased linearly (0.1041 MPa°Ch^?1, the slope of the relationship between ψ _b(50) and supra-optimal T _s) with T when T _s increased above T _o and finally reached to zero at T _c. The T _b and T _o values were not influenced by ψ, but T _c value decreased (from 47.2 for zero to 43.5 °C for ? 0.36 MPa) at supra-optimal T _s as a result of the effect of ψ on GR. Based on our findings, this model (as a predictive tool) and or the estimated parameter values in this study can easily be used in sesame SG simulation models to quantitatively characterize the physiological status of sesame seed populations at different T _s and ψ _s.     

Cryopreservation of <Emphasis Type="Italic">Citrus</Emphasis> anthers in the National Crop Genebank of China

Genebank conservation of pollen is valuable because it makes genetic resources immediately available for use in breeding programs. In the case of Citrus species, conserved anthers or pollen can be easily transported and used to develop new varieties with pathogen resistance and desirable quality and yield traits. The aim of this study was to develop and improve air-desiccation cryopreservation protocols for Citrus cavaleriei and Citrus maxima anthers in genebanks. In the current study, warming, rehydration, and in vitro germination conditions were optimized to achieve high levels of in vitro germination in Citrus pollen for ten cultivars after liquid nitrogen (LN) exposure. The optimal warming, rehydration, and in vitro germination medium formulations affected the germination levels after pollen cryopreservation, with species- and cultivar-dependent effects. The Citrus anthers were dehydrated to the moisture content of 5–14% before LN exposure and warmed at 25 (cryopreserved Citrus anthers with a moisture content of lower than 10%) or 37°C (a moisture content of 10% or higher), then rehydrated, and cultured on medium with 150-g L^?1 sucrose, 0.1-g L^?1 boric acid, 1.0-g L^?1 calcium nitrate, 0.1-g L^?1 potassium nitrate, 0.3-g L^?1 magnesium sulfate, and 10-g L^?1 agar. After 2 yr of storage, in vitro germination levels of Citrus pollen after cryopreservation were significantly higher (> 22% for all ten cultivars) than those of samples that were stored at 4°C (0%). In vitro germination levels of pollen from six of ten cultivars after cryopreservation remained relatively high after 2 yr of storage (38–93%). The highest viability of 93% was obtained for C. cavaleriei ‘2–3’. The methods identified in the current study could be used to cryopreserve C. cavaleriei and C. maxima anthers.     

Effects of drought stress on seed germination of ornamental sunflowers

The response to drought stress on germination was investigated on three hybrids of ornamental sunflower, ‘Hadar’, ‘Pazit’, and ‘Zohar’. Different levels of water potential [Ψ: 0.0 (control), ?0.15, ?0.30, ?0.45, ?0.60, ?0.75, and ?0.90 MPa] were adopted using polyethylene glycol-6000 (PEG6000) at four germination temperatures (15, 20, 25, and 30 °C). Final germination percentage, mean germination time, germination index, germination rate index, and germination stress tolerance index were used to evaluate the genotype response to PEG-induced water stress. Shoot and root length and fresh and dry weight were measured on seeds germinated at 20 °C under the different levels of water potentials. During germination, the three ornamental sunflowers showed to be more sensitive to suboptimal temperature than to supraoptimal. Decreasing water potential of imbibition solution progressively inhibited and delayed seed germination. Among cultivars, ‘Hadar’ and ‘Pazit’ performed better at temperature lower than 30 °C. ‘Zohar’ showed a lower sensitivity to PEG-induced water stress at all temperature conditions. Water stress during seed germination depressed the following seedling growth under favourable conditions. As a result, shoot and root length and fresh and dry weight was significantly lower in seedlings from seed germinated at ψ ≤ 0.45 MPa.     

Effect of cold stratification on seed germination in <Emphasis Type="Italic">Solidago</Emphasis> × <Emphasis Type="Italic">niederederi</Emphasis> (<Emphasis Type="Italic">Asteraceae</Emphasis>) and its parental species

In this study, we investigated the influence of cold stratification on seed germination in S. × niederederi, a hybrid between the North American S. canadensis and the European S. virgaurea, using fruit samples collected in 2016 in Poland. We aimed to test the hypothesis that the low temperature exposure decreases the final percentage and speed of seed germination in the hybrid and its parental species. For each species, sets of 100 achenes in three replications were mixed with dry sand and stored in Petri dishes in darkness for 12 weeks, at ?18 °C and?+?4 °C, and?+?25 °C. The seeds were incubated for 21 d at room temperature (+25 °C), under the 12 h photoperiod (630 lx). We showed a lack of significant differences in: (i) the final percentage of germinated seeds of studied species stored at the same conditions, (ii) the final percentage of germinated seeds between the applied stratification conditions in the hybrid and its parental species, and (iii) the mean values of Timson’s index, mean germination time, and coefficient of velocity of germination between the stratification conditions in each species. The statistically significant inter-specific differences in the mean germination time parameter after the +25 °C treatment suggest that the seeds of S. × niederederi are able to germinate faster than the seeds of its parental species. However, to improve our knowledge of naturalization and invasion abilities of S. × niederederi by sexual reproduction, the seed germination and seedling survival of the hybrid should be tested in the field.     

Mung bean (<Emphasis Type="Italic">Vigna radiata</Emphasis>) cultivars mediated oviposition preference and development of <Emphasis Type="Italic">Callosobruchus chinensis</Emphasis> (Coleoptera: Chrysomelidae: Bruchinae)

The Azuki bean weevil, Callosobruchus chinensis (L.), is a destructive pest of stored mung bean [Vigna radiata (L.) Wilczek] as well as other leguminous seeds. The development of resistant seeds to manage this pest is of current great interest to plant breeders. In this study, we investigated the oviposition preference and development of C. chinensis on two susceptible mung bean cultivars (Seonhwa and Gyeongseon) and one previously reported resistant cultivar (Jangan), compared to the susceptible cowpea (Vigna unguiculata L.), cultivar (Yeonbun) using both multiple-choice and no-choice tests. In addition, the development of C. chinensis was also examined at four constant temperatures (20, 25, 30, and 35 °C). Both tests found cowpea to be the most suitable seed for oviposition. Total developmental time from oviposition to adult emergence ranged from 27.01 to 38.2 days, being shortest on cowpea and longest on the mung bean, cv. Jangan. However, no successful development of C. chinensis larvae on mung bean, cv. Jangan, occurred at any temperature. The highest rate of adult emergence and the longest adult longevity both occurred on cowpea and certain mung bean cultivars (Seonhwa and Gyeongseon), with the dramatic exception of cv. Jangan. These results suggest that the higher preference and performance of C. chinensis on cowpea (3.3 egg/seed) and least on mung bean, cv. Jangan (0.4 egg/seed). This information may facilitate the exploration of resistant genetic materials and chemicals associated with seeds for successful breeding. Further studies should examine the chemicals associated with mung bean cultivars and its resistant mechanism to develop a control method against bruchines.     

Effect of Temperature on Development and Reproduction of <Emphasis Type="Italic">Epilachna dodecastigma</Emphasis> (Wied.) (Coleoptera: Coccinellidae)

The effect of five constant temperatures of 21, 24, 27, 30 and 33 °C on adult life span, reproduction, oviposition behavior and larval developmental time of a bitter gourd inhabited coleopteran insect Epilachna dodecastigma (Wied.) (Coccinellidae) was determined in laboratory conditions under 70 ± 5 % relative humidity and a photoperiod of 12 L : 12 D. Larval developmental time of E. dodecastigma decreased as temperature increased from 21 to 33 °C. Life table data revealed that overall mortality was lowest at 27 °C and highest at 21 °C. Females lived longer than males at all temperatures; but longevity decreased with increase in temperature. Pre-oviposition period decreased significantly with increase in temperature up to 27 °C and thereafter increased at a slower rate; whereas oviposition period decreased significantly with increase in temperature. Fecundity and egg viability increased significantly with an increase in temperature up to 27 °C and thereafter decreased at a slower rate. The intrinsic rate of increase (r _m ) was 0.1703, 0.1984, 0.2235, 0.2227 and 0.2181 day^?1 at 21, 24, 27, 30 and 33 °C, respectively. The net reproductive rate and finite rate of increase was highest at 27 °C (R _o  = 112.05; λ = 1.4233) and lowest at 21 °C (R _o  = 51.23; λ = 1.2581).     

Effect of short-term heat exposure on physiological traits of indica rice at grain-filling stage

Heat stress impacts the quantity and quality of rice grains, particularly during grain-filling stage needed for grain development. In this study, the effect of short heat stress (42 °C, 30 min) on indica rice plants at the grain-filling stage (dough grain stage) was found by determining their physiological and growth traits F_v/F_m, ?F/F_m′, chlorophyll content, leaf water potential (LWP), membrane stability, relative leaf area (RLA), relative plant height (RPH), total grain weight per panicle (TGW) and 1000 GW. Thai economic rice cvs. KDML105 and Pathumthani 1 (PTT1) were compared to the heat-tolerant rice cultivars N22 and Dular and to the heat-sensitive rice cultivar IR64. The results showed that short heat stress exhibited effects on physiology and growth greater than the control (35 °C, 30 min) by reducing of F_v/F_m, ?F/F_m′, chlorophyll content, LWP, membrane stability and RLA. This result impacted the TGW and 1000 GW. A higher reduction of physiological traits was shown in IR64, followed by KDML105. In contrast, N22 and Dular were minimally affected by heat stress and were able to adapt and recover based on their grain weight that exhibited less of an effect. PTT1 was also impacted by heat stress similarly to Dular. Thus, short heat stress affected the physiological parameters of five rice cultivars at the dough grain stage. In addition, the five indica rice cultivars were classified into three groups: (1) the heat-tolerant group (N22, Dular and PTT1), (2) the moderately heat-tolerant group (KDML105), and (3) the heat-sensitive group (IR64) by PC-ORD program at 50% of similarity of the 13 physiological traits.     

Purification and partial characterization of phytase from rice bean (<Emphasis Type="Italic">Vigna umbellata</Emphasis> Thunb.) germinated seeds

Rice bean (Vigna umbellata Thunb.) phytase activity increased during germination and reached maximum at 72 h. The phytate content in seeds decreased with increase in germination time. Phytase was purified 32 fold from 72-h germinated cotyledons with final specific activity 2.22 U/mg. Native PAGE revealed a single band. On SDS PAGE, it revealed two bands with molecular mass 66 and 44 kDa. The native molecular mass was 110 kDa on size exclusion chromatography. The A₂₈₀/₂₆₀ ratio was 1.88. When the enzyme was excited at 295 nm, the emission maximum was observed at 330 nm. The FTIR results suggest that Lys, Tyr, Phe, Trp, Ser, Gln and Asn residues on the enzyme’s surface. The enzyme was stored at 4 °C, showed 12 % residual activity on 35th day which was improved to 53.6 and 65.7 %, respectively in the presence of additives ascorbic acid and acetaminophen. The optimum pH and temperature of enzyme were 4.0 and 40 °C, respectively. The energy of activation was 32.2 kJ/mol. The values of K _m and V _max were 0.197 mM and 2.35 μmol/min/mg protein, respectively with sodium phytate as substrate. Phytase showed broad substrate specificity. The k _cat/K _m ratio was the highest for sodium phytate.     

High-yield expression,purification, characterization,and structure determination of tag-free <Emphasis Type="Italic">Candida utilis</Emphasis> uricase

We report the successful high-yield expression of Candida utilis uricase in Escherichia coli and the establishment of an efficient three-step protein purification protocol. The purity of the recombinant protein, which was confirmed to be C. utilis uricase by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer analysis, was >98% and the specific activity was 38.4 IU/mg. Crystals of C. utilis uricase were grown at 18°C using 25% polyethylene glycol 3350 as precipitant. Diffraction by the crystals extends to 1.93 Å resolution, and the crystals belong to the space group P2₁2₁2₁ with unit cell parameters a?=?69.16 Å, b?=?139.31 Å, c?=?256.33 Å, and α?=?β?=?γ?=?90°. The crystal structure of C. utilis uricase shares a high similarity with other reported structures of the homologous uricases from other species in protein database, demonstrating that the three-dimensional structure of the protein defines critically to the catalytic activities.     

Pistil-function breakdown in a new <Emphasis Type="Italic">S</Emphasis>-allele of European pear, <Emphasis Type="Italic">S</Emphasis><Subscript><Emphasis Type="Italic">21</Emphasis></Subscript>°, confers self-compatibility

European pear exhibits RNase-based gametophytic self-incompatibility controlled by the polymorphic S-locus. S-allele diversity of cultivars has been extensively investigated; however, no mutant alleles conferring self-compatibility have been reported. In this study, two European pear cultivars, ‘Abugo’ and ‘Ceremeño’, were classified as self-compatible after fruit/seed setting and pollen tube growth examination. S-genotyping through S-PCR and sequencing identified a new S-RNase allele in the two cultivars, with identical deduced amino acid sequence as S ₂₁ , but differing at the nucleotide level. Test-pollinations and analysis of descendants suggested that the new allele is a self-compatible pistil-mutated variant of S ₂₁ , so it was named S ₂₁ °. S-genotypes assigned to ‘Abugo’ and ‘Ceremeño’ were S ₁₀ S ₂₁ ° and S ₂₁ °S ₂₅ respectively, of which S ₂₅ is a new functional S-allele of European pear. Reciprocal crosses between cultivars bearing S ₂₁ and S ₂₁ ° indicated that both alleles exhibit the same pollen function; however, cultivars bearing S ₂₁ ° had impaired pistil-S function as they failed to reject either S ₂₁ or S ₂₁ ° pollen. RT-PCR analysis showed absence of S ₂₁ °-RNase gene expression in styles of ‘Abugo’ and ‘Ceremeño’, suggesting a possible origin for S ₂₁ ° pistil dysfunction. Two polymorphisms found within the S-RNase genomic region (a retrotransposon insertion within the intron of S ₂₁ ° and indels at the 3′UTR) might explain the different pattern of expression between S ₂₁ and S ₂₁ °. Evaluation of cultivars with unknown S-genotype identified another cultivar ‘Azucar Verde’ bearing S ₂₁ °, and pollen tube growth examination confirmed self-compatibility for this cultivar as well. This is the first report of a mutated S-allele conferring self-compatibility in European pear.     

Expression of ethylene biosynthetic and signaling genes in relation to ripening of banana fruit after cold storage

Banana fruit are highly sensitive to chilling injury (CI), while the effect of different degrees of CI on the subsequent fruit ripening is largely unknown. In the present work, ripening characteristic of banana fruit after storage at 7 °C for 3 days or for 8 days, and expression levels of eight genes associated with ethylene biosynthetic and signaling, including MaACS1, MaACO1, MaERS1, MaERS3, and MaEIL1–4, were investigated. The results showed that banana fruit stored at 7 °C for 8 days exhibited more severe chilling symptoms than those at 7 °C for 3 days. Compared with banana fruit stored at 7 °C for 8 days, which showed abnormal ripening, more decrease in fruit firmness, while higher increase in ethylene production and hue angle were observed in banana fruit stored at 7 °C for 3 days, which could ripening normally. Moreover, gene expression profiles during ripening revealed that ethylene biosynthetic and signaling genes were differentially expressed in peel and pulp of banana fruit after storage at 7 °C for 3 days and 7 °C for 8 days. In the peel of fruit storage at 7 °C for 3 days, expression levels of MaACS1, MaACO1, MaEIL1, and MaEIL2 increased remarkably while MaERS3, MaEIL1, and MaEIL4 were enhanced in the fruit after storage at 7 °C for 8 days. In the pulp, with the exception of MaACO1 and MaERS3, expression levels of other genes did not exhibit a significant difference, between the banana fruit storage at 7 °C for 3 days and 7 °C for 8 days. Taken together, our results suggest that differential expression of ethylene biosynthetic and signaling genes such as MaERS3, MaACO1, and MaEIL2, may be related to ripening behavior of banana fruit with different degrees of CI after cold storage.     

Biology and Thermal Requirements of <Emphasis Type="Italic">Fopius arisanus</Emphasis> (Sonan, 1932) (Hymenoptera: Braconidae) Reared on <Emphasis Type="Italic">Ceratitis capitata</Emphasis> Eggs (Wiedemann) (Diptera: Tephritidae)

Fopius arisanus (Sonan) is a solitary parasitoid of eggs and the first instar larvae of Tephritidae. Due to the occurrence of Ceratitis capitata (Wiedemann) in various regions and under several climatic conditions, this study aimed to evaluate the effect of different temperatures on the embryonic development (egg–adult) and determine thermal requirements and the number of annual generations F. arisanus on eggs of C. capitata. In the laboratory, eggs of C. capitata (24 h) were submitted to parasitism of F. arisanus during 6 h. Later, the eggs were placed in plastic containers (50 mL) (50 eggs/container) on a layer of artificial diet and packed in chambers at temperatures 15, 18, 20, 22, 25, 28, 30, and 32 ± 1°C, RH 70 ± 10%, and a photophase of 12 h. The largest number of offspring, emergence rate, and weight of adults of F. arisanus were observed at 25°C. The highest sex ratios (sr > 0.75) were recorded at 15 and 18°C, being statistically higher than the temperatures 20°C (0.65), 22°C (0.64), 25°C (0.65), 28°C (0.49), and 30°C (0.47). At 32°C, there was no embryonic development of F. arisanus. The egg–adult period was inversely proportional to temperature. Based on the development of the biological cycle (egg–adult), the temperature threshold (T _t) was 10.3°C and thermal constant (K) of 488.34 degree-days, being the number of generations/year directly proportional to the temperature increase. The data show the ability of F. arisanus to adapt to different thermal conditions, which is important for biological control programs of C. capitata.     

Temperature-induced changes in treatment efficiency and microbial structure of aerobic granules treating landfill leachate

This paper investigates the effect of temperature on nitrogen and carbon removal by aerobic granules from landfill leachate with a high ammonium concentration and low concentration of biodegradable organics. The study was conducted in three stages; firstly the operating temperature of the batch reactor with aerobic granules was maintained at 29 °C, then at 25 °C, and finally at 20 °C. It was found that a gradual decrease in operational temperature allowed the nitrogen-converting community in the granules to acclimate, ensuring efficient nitrification even at ambient temperature (20 °C). Ammonium was fully removed from leachate regardless of the temperature, but higher operational temperatures resulted in higher ammonium removal rates [up to 44.2 mg/(L h) at 29 °C]. Lowering the operational temperature from 29 to 20 °C decreased nitrite accumulation in the GSBR cycle. The highest efficiency of total nitrogen removal was achieved at 25 °C (36.8 ± 10.9 %). The COD removal efficiency did not exceed 50 %. Granules constituted 77, 80 and 83 % of the biomass at 29, 25 and 20 °C, respectively. Ammonium was oxidized by both aerobic and anaerobic ammonium-oxidizing bacteria. Accumulibacter sp., Thauera sp., cultured Tetrasphaera PAO and Azoarcus–Thauera cluster occurred in granules independent of the temperature. Lower temperatures favored the occurrence of denitrifiers of Zooglea lineage (not Z. resiniphila), bacteria related to Comamonadaceae, Curvibacter sp., Azoarcus cluster, Rhodobacter sp., Roseobacter sp. and Acidovorax spp. At lower temperatures, the increased abundance of denitrifiers compensated for the lowered enzymatic activity of the biomass and ensured that nitrogen removal at 20 °C was similar to that at 25 °C and significantly higher than removal at 29 °C.     

Seed germinability and longevity influences regeneration of <Emphasis Type="Italic">Acacia gerrardii</Emphasis>

Acacia gerrardii is the only native tree species of the Kuwaiti desert ecosystem. However, anthropogenic disturbances and harsh arid climate have contributed towards the disappearance of this keystone species from its habitat. In this study, effects of different seed pretreatments to break dormancy, water entry pathway, and ecology (seasonal timing) of dormancy loss and germination of A. gerrardii were investigated. Effects of mechanical scarification, hot water treatment (30 s, 1, 2, and 5 min), and concentrated acid scarification (10, 20, and 30 min) on germination percentage and rate (time to 50% germination and final germination) were also examined. Pretreatment with mechanical scarification produced the highest germination in the least time and 20 °C, 40% RH with 12 h of light (2370 Lux) were found to provide the best germination environment. Seeds were rapidly aged at 60% RH and 45 or 50 °C to determine longevity, and the results were analyzed using probit analysis. Times taken for viability of A. gerrardii seeds aged at 45 and 50 °C to fall to 50% (p₅₀) were 38.6 and 9.3 days, respectively, and therefore the seeds can be considered to have medium longevity. Experiments to find the water entry pathway in A. gerrardii indicated that the micropyle region was the primary point of water entry into the seed. Seed burial experiments indicated that though seed retention decreased over time, there was no significant decrease in number of viable seeds after 31 weeks. The findings of this study are important to nursery managers, seed banks, and those involved in conservation and restoration activities.     

Droplet-vitrification cryopreservation of <Emphasis Type="Italic">in vitro</Emphasis>-grown shoot tips of grapevine (<Emphasis Type="Italic">Vitis</Emphasis> spp.)

An efficient and broad-spectrum protocol for cryopreservation of Vitis spp. shoot tips by droplet-vitrification is reported. Shoot tips (1.0 mm) containing 5–6 leaf primordia (LPs) were precultured for 3 d with a preculture medium containing 0.3 M sucrose, 0.16 μM glutathione, and 0.14 μM ascorbic acid. Precultured shoot tips were treated for 20 min at 24°C with a loading solution composed of 2 M glycerol and 0.4 M sucrose, followed by exposure at 0°C to half-strength plant vitrification solution 2 (PVS2) for 30 min, and then full-strength PVS2 for 50 min. Dehydrated shoot tips were transferred into 2.5-μL PVS2 carried on aluminum foil, prior to a direct immersion in liquid nitrogen. With this method, an average shoot regrowth level of 50.5% was obtained from cryopreserved shoot tips in six V. vinifera genotypes (three wine cultivars, two table cultivars, and one rootstock) and two V. pseudoreticulata genotypes. Vegetative growth of the regenerants recovered from cryopreservation, significantly increased as the number of subculture cycles increased and was greater than the control after the third subculture following cryopreservation. Inter-simple sequence repeats (ISSR) and random amplification of polymorphic DNA (RAPD) analyses did not detect any polymorphic loci in the plants of V. vinifera L. cv. ‘Cabernet Sauvignon’ from cryopreserved shoot tips compared to the original cultures. This droplet-vitrification cryopreservation method provides a technical platform to set up cryobanks of Vitis spp.     

Population Growth Parameters of Rose Aphid, <Emphasis Type="Italic">Macrosiphum rosae</Emphasis> (Hemiptera: Aphididae) on Different Rose Cultivars

The rose aphid, Macrosiphum rosae (L.), is one of the most important pests on rose plants (Rosa spp.) with a worldwide distribution. As resistance indices, the development, survivorship, and reproduction of this aphid were evaluated on 10 rose cultivars, including Bella Vita, Cool Water, Dolce Vita, Maroussia, Orange Juice, Pinkpromise, Roulette, Tea, Valentine, and Persian Yellow in laboratory at 25?±?1°C, 65?±?5% relative humidity, and photoperiod of 16:8 (L/D)?h. Rose aphid successfully survived on all 10 rose cultivars, although mortality rate was higher on Tea and Bella Vita. The number of offspring per female differed significantly among the tested rose cultivars, and ranged from 9.2 on Tea to 38.7 nymphs on Orange Juice. Population growth parameters were significantly affected by rose cultivars. The longest mean generation time (T) was observed on Bella Vita (14.8 days) and Tea (14.7 days) and the shortest on Orange Juice (10.0 days). The net reproductive rate (R ₀ ) ranged from 6.9 on Tea to 33.2 nymphs on Orange Juice cultivar. Correspondingly, the highest value of intrinsic rate of increase (r _m ) was observed on Orange Juice (0.348 day^?1) and lower values on Tea (0.131 day^?1) followed by Bella Vita (0.154 day^?1). Cluster analysis of all the measured parameters of rose aphid on different rose cultivars revealed that Tea and Bella Vita were relatively resistant to M. rosae. These findings could be useful in developing an integrated pest management (IPM) program for this aphid in urbanized areas and commercial rose potting.     

Differences in neurochemical profiles of two gadid species under ocean warming and acidification

Background

Exposure to future ocean acidification scenarios may alter the behaviour of marine teleosts through interference with neuroreceptor functioning. So far, most studies investigated effects of ocean acidification on the behaviour of fish, either isolated or in combination with environmental temperature. However, only few physiological studies on this issue were conducted despite the putative neurophysiological origin of the CO₂-induced behavioural changes. Here, we present the metabolic consequences of long-term exposure to projected ocean acidification (396–548 μatm PCO₂ under control and 915–1272 μatm under treatment conditions) and parallel warming in the brain of two related fish species, polar cod (Boreogadus saida, exposed to 0 °C, 3 °C, 6 °C and 8 °C) and Atlantic cod (Gadus morhua, exposed to 3 °C, 8 °C, 12 °C and 16 °C). It has been shown that B. saida is behaviourally vulnerable to future ocean acidification scenarios, while G. morhua demonstrates behavioural resilience.

Results

We found that temperature alters brain osmolyte, amino acid, choline and neurotransmitter concentrations in both species indicating thermal responses particularly in osmoregulation and membrane structure. In B. saida, changes in amino acid and osmolyte metabolism at the highest temperature tested were also affected by CO₂, possibly emphasizing energetic limitations. We did not observe changes in neurotransmitters, energy metabolites, membrane components or osmolytes that might serve as a compensatory mechanism against CO₂ induced behavioural impairments. In contrast to B. saida, such temperature limitation was not detected in G. morhua; however, at 8 °C, CO₂ induced an increase in the levels of metabolites of the glutamate/GABA-glutamine cycle potentially indicating greater GABAergic activity in G.morhua. Further, increased availability of energy-rich substrates was detected under these conditions.

Conclusions

Our results indicate a change of GABAergic metabolism in the nervous system of Gadus morhua close to the optimum of the temperature range. Since a former study showed that juvenile G. morhua might be slightly more behaviourally resilient to CO₂ at this respective temperature, we conclude that the observed change of GABAergic metabolism could be involved in counteracting OA induced behavioural changes. This may serve as a fitness advantage of this respective species compared to B. saida in a future warmer, more acidified polar ocean.