Coffee volatiles induced after mechanical injury and beetle herbivory attract the coffee berry borer and two of its parasitoids

The coffee berry borer (CBB), Hypothenemus hampei (Ferrari), is the most important insect pest of coffee worldwide. In this study, we used headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry to sample and identify volatile compounds from Robusta coffee berries, Coffea canephora Pierre ex Froehner, infested with CBB and with mechanical damage. Furthermore, we evaluated the behavioral responses of the CBB and two of its parasitoids, Prorops nasuta Waterstone and Phymastichus coffea LaSalle, to three selected coffee volatile compounds in a Y-tube olfactometer. We found in the effluvia of red coffee berry compounds not previously reported for this coffee species. Our results show that Robusta coffee berries release induced volatiles either by insect herbivory or by mechanical damage. Small amount of butyl acetate, unknown compound 2, α-longipinene, longiborneol and longiborneol acetate are produced only in infested coffee berries fruits. Quantitatively, nine compounds account for the difference between healthy berries, infested, or mechanically damaged berries. Trans-ocimene, 4,8-dimethyl-3,7-nonadien-2-ol, α-copaene and kaurene increased amount levels in infested berries, while amount of methyl salicylate and linalool increased in mechanically damaged coffee berries. The olfactometric bioassays showed that CBB females and its two parasitoids were attracted to methyl salicylate. In addition, H. hampei and P. nasuta were attracted to linalool, and P. nasuta and P. coffea were attracted to trans-ocimene.     

Song frequency correlates with latitude and individual body size in the cicada <Emphasis Type="Italic">Mogannia formosana</Emphasis> Matsumura (Hemiptera: Cicadidae)

Analyses of acoustic variation between and within populations can help to clarify the evolution and diversification of cicada calling songs. In this study, we analyzed the acoustic variation in the calling song of Mogannia formosana within Taiwan and between Taiwan and Green Island to assess the effects of geographic locations. Furthermore, chorusing males in the Green Island population were recorded and collected from the same habitat site during the same time period to investigate the relationship between individual body size and the acoustic features of calling songs. Among populations of M. formosana, we found that most of the acoustic variation in M. formosana calling songs was associated with frequency parameters, in which six frequency parameters changed significantly with latitude on the island of Taiwan. In contrast, temporal parameters, which were associated with principal components corresponding to less acoustic variation than that of the first principal component, were also found to be significant among populations but did not show consistent trends of difference with latitude. However, the geographically isolated Green Island population exhibited the lowest number of short echemes in segment B, which is the diagnostic structure employed to separate M. formosana from other M. species. This finding suggests that the temporal pattern of segment B in the calling songs of M. formosana might be important for both population differentiation and interspecific recognition. In a chorus of the Green Island population, the sound frequency of the last short echeme was found to be significantly correlated with individual body size. The possible role played by sexual selection in shaping sound frequency as a result of its reliable association with body size was discussed. We suggest that, in comparison with temporal elements, the frequency elements of segment B in calling songs of M. formosana in chorus can serve as a more reliable indicator of body size for female mate choice.     

Circadian rhythm of activity as a factor of reproductive isolation in closely related species of sand-desert tenebrionid beetles (Coleoptera,Tenebrionidae)

The results of long-term studies of the circadian rhythms of activity in Trigonoscelis gigas and T. sublaevicollis, typical and abundant representatives of the tenebrionid fauna in the Kara Kum desert are given. T. gigas and T. sublaevicollis are omni-seasonal species: their adults are observed on the sand surface from spring till late autumn. They spend their resting period in soil. Adults of both species are similar in morphology, ecology, and behavior, strongly differing in the circadian rhythm pattern. T. gigas are active only at twilight, whereas T. sublaevicollis are characterized by nocturnal activity. Therefore, inhabiting the same habitat, T. gigas and T. sublaevicollis never meet each other under natural conditions, independently of the season and weather. Hence, the circadian rhythm of activity can serve as a reliable mechanism of reproductive isolation in closely related tenebrionid species.     

Comparison of bioethanol production from cultivated versus wild <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis> and <Emphasis Type="Italic">Gracilaria gigas</Emphasis>

The seaweed genus Gracilaria is a potential candidate for the production of bioethanol due to its high carbohydrate content. Gracilaria is abundant throughout the world and can be found in both wild and cultivated forms. Differences in the ecological factors such as temperature, salinity, and light intensity affecting wild and cultivated specimens may influence the biochemical content of seaweeds, including the carbohydrate content. This study aimed to investigate the proximate composition and potential bioethanol production of wild and cultivated G. gigas and G. verrucosa. Bioethanol was produced using separate hydrolysis fermentation (SHF), employing a combination of enzymatic and acid hydrolysis, followed by fermentation with Saccharomyces cerevisiae ATCC 200062. The highest carbohydrate content was found in wild G. gigas. The highest galactose and glucose contents (20.21 ± 0.32 and 9.70 ± 0.49 g L^?1, respectively), as well as the highest production of bioethanol (3.56 ± 0.02 g L^?1), were also found in wild G. gigas. Thus, we conclude that wild G. gigas is the most promising candidate for bioethanol production. Further research is needed to optimize bioethanol production from wild G. gigas. Domestication of wild G. gigas is a promising challenge for aquaculture to avoid overexploitation of this wild seaweed resource.     

Home range size in two darkling beetle species (Coleoptera,Tenebrionidae) from Southern Kazakhstan

The repeated collecting of marked darkling beetle species, including the diurnal species Tentyria gigas and the nocturnal species Cyphogenia gibba from Kazakhstan demonstrated that their home ranges constituted 457 and 360 m², respectively. More than 500 beetles from an area of 2.5 ha were marked by engraving numbers on their elytra. Beetles were observed during two seasons at a distance covering 2 km from the site of release. Sex was determined in a quarter of all the beetles. In females and males of T. gigas, the home range constituted 700 and 552 m², respectively; in C. gibba, 599 and 343 m², respectively. Small home ranges of these species can be explained by the presence of numerous predators, occasional death of beetles under cattle’s hoofs, and a great number and high density of shelters.     

Dynamics of periodical cicada chorus centers (Homoptera: Cicadidae:Magicicada)

Mating aggregations of three species of periodical cicadas were monitored during the emergence of Brood XIX at a 16-ha study site in northwest Arkansas, May–June 1985. Magicicada tredecassiniappeared first and formed the most choruses. M. tredecimand M. tredeculachoruses formed next, and M. tredeculachoruses outnumbered those of M. tredecim.Of the 268 choruses seen, 84% were composed of M. tredecassini. M. tredecassiniwere often found chorusing in the same trees with the other two species. Such multispecies mating aggregations apparently are unique to periodical cicadas. Choruses were dynamic with respect to their locations and durations. Initially, choruses were located near areas of high cicada emergence densities. One week later, cicadas chorused in trees throughout the forest and at the forest edge. Many choruses were seen only once at a location. Although cicadas chorused for almost 4 weeks, individual choruses persisted only approximately 8 days, on average. Sound intensities under chorus centers ranged from 50 to 80 dh and were correlated with arena sizes during times of peak chorus activity. No distinct habitat preferences of the three species were observed, however, the tree species used by chorusing cicadas differed among the species.     

Intraspecific Variation in Mitogenomes of Five <Emphasis Type="Italic">Crassostrea</Emphasis> Species Provides Insight into Oyster Diversification and Speciation

A large number of Crassostrea oysters are found in Asia-Pacific. While analyses of interspecific variation have helped to establish historical relationships among these species, studies on intraspecific variation are necessary to understand their recent evolutionary history and current forces driving population biology. We resequenced 18 and analyzed 31 mitogenomes of five Crassostrea species from China: Crassostrea gigas, Crassostrea angulata, Crassostrea sikamea, Crassostrea ariakensis, and Crassostrea hongkongensis. Our analysis finds abundant insertions, deletions, and single-nucleotide polymorphisms in all species. Intraspecific variation varies greatly among species with polymorphic sites ranging from 54 to 293 and nucleotide diversity ranging from 0.00106 to 0.00683. In all measurements, C. hongkongensis that has the narrowest geographic distribution exhibits the least sequence diversity; C. ariakensis that has the widest distribution shows the highest diversity, and species with intermediate distribution show intermediate levels of diversity. Low sequence diversity in C. hongkongensis may reflect recent bottlenecks that are probably exacerbated by human transplantation. High diversity in C. ariakensis is likely due to divergence of northern and southern China populations that have been separated without gene flow. The significant differences in mitogenome diversity suggest that the five sister species of Crassostrea have experienced different evolutionary forces since their divergence. The recent divergence of two C. ariakensis populations and the C. gigas/angulata species complex provides evidence for continued diversification and speciation of Crassostrea species along China’s coast, which are shaped by unknown mechanisms in a north–south divide.     

Nucleotide Diversity of the Coding and Promoter Regions of <Emphasis Type="Italic">DREB1D</Emphasis>, a Candidate Gene for Drought Tolerance in <Emphasis Type="Italic">Coffea</Emphasis> Species

Climate change is posing a major challenge to coffee production worldwide leading to a need for the development of coffee cultivars with increased drought tolerance. In several plant species, the use of DREB genes in crop improvement has achieved promising results to desiccation tolerance engineering. Recent studies reported CcDREB1D specific patterns of expression in Coffea canephora and functional evidence of this gene involvement in drought stress responses. However, knowledge on natural diversity of this gene is largely unknown. In this context, this study aimed at evaluating the sequence variability of the DREB1D gene in several Coffea genotypes. Nucleotide variation in promoters and coding regions of this gene were evaluated in a population consisting of 38 genotypes of C. canephora, C. arabica and C. eugenioides, most of them characterized by different phenotypes (tolerance vs. susceptibility) in relation to drought. The genetic diversity of the loci revealed different haplotypes for the promoter and coding regions. In particular, our findings suggest association between drought tolerance and the genetic variations on DREB1D promoter regions, but not with those from its corresponding coding regions. Gene expression studies revealed up-regulated expression of DREB1D gene upon drought mainly in leaves of drought-tolerant clones of C. canephora, and in response to drought, high, and low temperatures in leaves of C. arabica, suggesting a key role of this gene in coffee responses to abiotic stress.     

Geographic patterns of genetic variation in nuclear and chloroplast genomes of two related oaks (<Emphasis Type="Italic">Quercus aliena</Emphasis> and <Emphasis Type="Italic">Q. serrata</Emphasis>) in Japan: implications for seed and seedling transfer

In this study, we assessed geographic patterns of genetic variations in nuclear and chloroplast genomes of two related native oaks in Japan, Quercus aliena and Q. serrata, in order to facilitate development of genetic guidelines for transfer of planting stocks for each species. A total of 12 populations of Q. aliena and 44 populations of Q. serrata were analyzed in this study. Genotyping of nuclear microsatellites in Q. aliena was done with only nine populations (n = 212) due to limited numbers of individuals in two populations, while all 12 populations (n = 89) were used in sequencing chloroplast DNA (cpDNA). In Q. serrata, 43 populations (n = 1032) were genotyped by nuclear microsatellite markers, while cpDNA of 44 populations (n = 350) was sequenced. As anticipated, geographic patterns detected in the variations of Q. aliena’s nuclear genome and its chloroplast haplotype distribution clearly distinguished northern and southern groups of populations. However, those of Q. serrata were inconsistent. The geographic distribution of its chloroplast haplotypes tends to show the predicted differentiation between northern and southern lineages, but geographic signals in the genetic structure of its nuclear microsatellites are weak. Therefore, treating northern and southern regions of Japan as genetically distinct transferrable zones for planting stocks is highly warranted for Q. aliena. For Q. serrata, the strong NE-SW geographic structure of cpDNA should be considered.     

Reproductive Biology of <Emphasis Type="Italic">Leptocybe invasa</Emphasis> Fisher &amp; La Salle (Hymenoptera: Eulophidae)

Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae) is an invasive pest in Eucalyptus plantations around the world. The successful colonization of L. invasa is possibly related to its reproductive biology. The objective of this study was to examine the reproductive biology of L. invasa. In Guangxi Province, the sex ratio (proportion of female, 0.99) of L. invasa was female-dominant throughout the year based on natural and artificial infestation. This result was similar to the ratios observed for other geographic populations in China, including those in Fujian (0.99), Guangdong (0.98), Hainan (0.95), Jiangxi (0.96), and Sichuan (0.99). The offspring sex ratio favored females. A large number of females emerged from the galls produced by females, with few males found. Galls on the petioles and midribs of Eucalyptus plants could be caused by newly emerged females with mature eggs. The lengths of the ovariole, spermatheca, common oviduct, and reproductive glands did not differ among L. invasa females, but their lateral oviducts showed differences from 0 to 42 h after emergence, indicating that this insect is proovigenic. These results could explain why L. invasa populations can rapidly increase in invaded areas.     

Functional convergence and phenotypic divergence in two specialist species of pine-associated ladybirds

Traditionally, convergent evolution has been considered to produce phenotypic similarity in independently evolved species. By contrast, recent studies have detected morphological divergence between species even in similar selective environments when different morphological traits combine to produce a specific functional output. However, it is still unclear whether a complex combination of non-morphological phenotypic traits, such as behavioural and life-history traits, can produce a similar performance in different species. In this study, I examined prey capture performance and related phenotypes in two sympatric ladybird species, Sospita oblongoguttata and Harmonia yedoensis, which specialize on the giant pine aphid, which is known to be elusive for ladybird hatchlings. In particular, I focused on egg size and proportion of trophic eggs in the clutch, since the amount of maternal investment per offspring can contribute to prey capture performance of ladybird hatchlings. Predation success of hatchlings against the giant pine aphid was higher in both S. oblongoguttata and H. yedoensis than in Harmonia axyridis, a generalist ladybird species that feeds on various kinds of aphid species in nature. Sospita oblongoguttata females, however, produced relatively larger eggs and in most clutches provided no trophic eggs, whereas H. yedoensis females produced smaller eggs and provided more trophic eggs per clutch. Moreover, hatchling morphology in H. yedoensis more closely resembled that of its congener, H. axyridis, than that of the more distantly related S. oblongoguttata, although like H. yedoensis, S. oblongoguttata predates on the elusive pine aphid. These results in two pine-associated specialist ladybirds indicate that divergent phenotypes can nonetheless have similar prey capture performance. In conclusion, this work demonstrates that the general ultimate function can be achieved by various mechanisms through convergence that operates at different level of life.     

Prolonged diapause of specialist seed-feeders makes predator satiation unstable in masting of<Emphasis Type="Italic"> Quercus crispula</Emphasis>

Quercus crispula (=Q. mongolica var. grosseserrata) is the predominant tree species in cool temperate, mixed broadleaf/conifer forests in northern Japan. We compared 11 years of data on acorn production in a population of Q. crispula, with data on seed-insect populations, to try to answer the following questions: (1) Does Q. crispula show a regular pattern of masting? (2) How long do principal seed predators remain in diapause? (3) How do the seed predators affect the pattern of predator satiation? Q. crispula showed a tendency to alternate bearing, with significant synchrony between individual trees. The principal acorn-feeding insects (Curculio spp. weevils), which infested 25%–70% of matured acorns, generally exhibited a prolonged diapause of 2 years. No significant negative relationship was found between the rate of injury by the weevils and the density of mature acorns, indicating that simple predator satiation fails due to the synchrony of the life-cycle of acorn-feeding insects and the periodical production of acorns. However, the rate of injury by the weevils was negatively correlated with the relative abundance of mature acorns to the number of weevil larvae that had matured 2 years previously. Thus, the proportion of sound acorns notably increased in a rich crop after a disturbance in alternate bearing. Prolonged diapause of specific seed predators is critical in determining the peak year of sound-seed production.     

Stridulatory Sound-Production and Its Function in Females of the Cicada Subpsaltria yangi

Acoustic behavior plays a crucial role in many aspects of cicada biology, such as reproduction and intrasexual competition. Although female sound production has been reported in some cicada species, acoustic behavior of female cicadas has received little attention. In cicada Subpsaltria yangi, the females possess a pair of unusually well-developed stridulatory organs. Here, sound production and its function in females of this remarkable cicada species were investigated. We revealed that the females could produce sounds by stridulatory mechanism during pair formation, and the sounds were able to elicit both acoustic and phonotactic responses from males. In addition, the forewings would strike the body during performing stridulatory sound-producing movements, which generated impact sounds. Acoustic playback experiments indicated that the impact sounds played no role in the behavioral context of pair formation. This study provides the first experimental evidence that females of a cicada species can generate sounds by stridulatory mechanism. We anticipate that our results will promote acoustic studies on females of other cicada species which also possess stridulatory system.     

Daily activity rhythm of desert omni-seasonal Tenebrionid beetles, <Emphasis Type="Italic">Trigonoscelis gigas</Emphasis> and <Emphasis Type="Italic">T. sublaevicollis</Emphasis> (Coleoptera,Tenebrionidae) in constant darkness and under alternating 1-hour pulses of light and darkness

In the Karakum Desert (Turkmenistan) the beetles Trigonoscelis gigas are only active in the morning and evening while T. sublaevicollis are strictly nocturnal, regardless of the season and weather. The daily activity rhythm of T. gigas and T. sublaevicollis was studied in the laboratory according to the following pattern: 5 days under a light-darkness cycle of 15: 9 h (LD 15: 9), then 10 days in constant darkness (DD), and then 10 more days under alternating 1-h pulses of light and darkness (LD 1: 1). The temperature was 25°C in all the modes. At LD 15: 9, beetles of both species maintained a 24-h period and a natural pattern of the activity rhythm. In DD, the circadian rhythm ran with a period of 23.5 ± 0.3 h (n = 40) in T. gigas and 23.6 ± 0.4 h (n = 40) in T. sublaevicollis. At DD, the morning and evening activity peaks of T. gigas merged to form a rhythm with only one peak. Under LD 1: 1, both T. gigas and T. sublaevicollis recovered a 24-h period of the rhythm, while the rhythm of T. gigas regained the two-peak structure. Our research confirmed the assumption of Tshernyshev (1980) about the 24-h period of the free-running endogenous rhythm and the distorting effect of constant conditions on this rhythm.     

Features of the reproductive cycle in females of the beaked redfish <Emphasis Type="Italic">Sebastes mentella</Emphasis> (Sebastidae)

The reproductive status of some categories of females of the beaked redfish Sebastes mentella has been determined as a result of the histological analysis. The histological characteristic of gonads of large nonreproductive females of redfish is presented for the first time. Gonads of mature females that skip spawning are studied. The cytomorphological characteristic of ovaries after larvae extrusion is specified. A broadened maturity scale for gonads of females of S. mentella supplemented with stages that characterize fish that skip spawning and large nonreproductive females is proposed.     

Reproductive features of <Emphasis Type="Italic">Upeneus margarethae</Emphasis> (Mullidae), a species recorded in the coastal zone of Vietnam for the first time

The species Upeneus margarethae described recently from the western Indian Ocean is recorded in the coastal zone of Vietnam for the first time. The reproductive features of the representatives of the species from southern Central Vietnam (Nha Trang Bay), Central Vietnam (Da Nang), and North Vietnam (Ha Long Bay) are studied. Based on the analysis of maturity stages of female’s ovaries and gonadosomatic index, the fishes spawn over the entire year in Central Vietnam, but the spawning interruption is observed in North Vietnam during a winter season. In Nha Trang Bay, 50% of females reach sexual maturity at the body length (FL) 9.8 cm. Based on histological structure of ovaries and frequency distribution of oocyte diameter before the spawning, the oogenesis is continuous. Anomalous oocyte structure is registered in 20% of the females. Egg morphology is described for 3 h after activation of ovulated oocytes. A comparison of reproductive parameters in U. margarethae and U. tragula is conducted.     

Sound waves increases the ascorbic acid content of alfalfa sprouts by affecting the expression of ascorbic acid biosynthesis-related genes

Previous studies have shown that sound wave treatment can affect the expression of plant genes and improve the growth. So, we investigated the ability of sound waves to increase AsA (l-ascorbic acid) content in alfalfa (Medicago sativa) sprouts in this study. Sprouts were exposed to a range of sound wave frequencies for two 1-h periods per day for various numbers of days. Most sound wave treated sprouts had a higher AsA content than untreated sprouts. In addition, the activity level of superoxide dismutase, an enzyme with potent antioxidative properties, was increased in sound wave-treated sprouts. The AsA content varied in response to sound wave treatment. Most processing conditions, including 500 and 1000 Hz, increased AsA content by 24–50%; however, some treatment conditions caused reduced AsA content during sprout growth. Furthermore, AsA content during sprout storage was increased by most sound wave treatment conditions, with 13–36% increases observed following 800 and 1000 Hz sound wave treatments compared to untreated sprouts. To investigate the mechanisms underlying changes in AsA content, we analyzed the expression levels of AsA biosynthesis-related genes. We found that several genes, including VTC1, VTC2, VTC4, GME, L-GalDH, GLDH, MDHAR, and DHAR1, displayed differential expression in response to sound wave treatment. Therefore, sound wave treatment may be a viable method for increasing the nutritional contents of sprouted vegetables.     

A preliminary integrated genetic map distinguishes every chromosome pair and locates essential genes related to abiotic adaptation of <Emphasis Type="Italic">Crassostrea angulata</Emphasis>/<Emphasis Type="Italic">gigas</Emphasis>

Background

The re-sequencing of C. angulata has revealed many polymorphisms in candidate genes related to adaptation to abiotic stress that are not present in C. gigas; these genes, therefore, are probably related to the ability of this oyster to retain high concentrations of toxic heavy metals. There is, in addition, an unresolved controversy as to whether or not C. angulata and C. gigas are the same species or subspecies. Both oysters have 20 metacentric chromosomes of similar size that are morphologically indistinguishable. From a genomic perspective, as a result of the great variation and selection for heterozygotes in C. gigas, the assembly of its draft genome was difficult: it is fragmented in more than seven thousand scaffolds.

Results

In this work sixty BAC sequences of C. gigas downloaded from NCBI were assembled in BAC-contigs and assigned to BACs that were used as probes for mFISH in C. angulata and C. gigas. In addition, probes of H3, H4 histone, 18S and 5S rDNA genes were also used. Hence we obtained markers identifying 8 out the 10 chromosomes constituting the karyotype. Chromosomes 1 and 9 can be distinguished morphologically. The bioinformatic analysis carried out with the BAC-contigs annotated 88 genes. As a result, genes associated with abiotic adaptation, such as metallothioneins, have been positioned in the genome. The gene ontology analysis has also shown many molecular functions related to metal ion binding, a phenomenon associated with detoxification processes that are characteristic in oysters. Hence the provisional integrated map obtained in this study is a useful complementary tool for the study of oyster genomes.

Conclusions

In this study 8 out of 10 chromosome pairs of Crassostrea angulata/gigas were identified using BAC clones as probes. As a result all chromosomes can now be distinguished. Moreover, FISH showed that H3 and H4 co-localized in two pairs of chromosomes different that those previously escribed. 88 genes were annotated in the BAC-contigs most of them related with Molecular Functions of protein binding, related to the resistance of the species to abiotic stress. An integrated genetic map anchored to the genome has been obtained in which the BAC-contigs structure were not concordant with the gene structure of the C. gigas scaffolds displayed in the Genomicus database.