Ecological genomics of tropical trees: how local population size and allelic diversity of resistance genes relate to immune responses, cosusceptibility to pathogens, and negative density dependence.

In tropical forests, rarer species show increased sensitivity to species-specific soil pathogens and more negative effects of conspecific density on seedling survival (NDD). These patterns suggest a connection between ecology and immunity, perhaps because small population size disproportionately reduces genetic diversity of hyperdiverse loci such as immunity genes. In an experiment examining seedling roots from six species in one tropical tree community, we found that smaller populations have reduced amino acid diversity in pathogen resistance (R) genes but not the transcriptome in general. Normalized R gene amino acid diversity varied with local abundance and prior measures of differences in sensitivity to conspecific soil and NDD. After exposure to live soil, species with lower R gene diversity had reduced defence gene induction, more cosusceptibility of maternal cohorts to colonization by potentially pathogenic fungi, reduced root growth arrest (an R gene-mediated response) and their root-associated fungi showed lower induction of self-defence (antioxidants). Local abundance was not related to the ability to induce immune responses when pathogen recognition was bypassed by application of salicylic acid, a phytohormone that activates defence responses downstream of R gene signalling. These initial results support the hypothesis that smaller local tree populations have reduced R gene diversity and recognition-dependent immune responses, along with greater cosusceptibility to species-specific pathogens that may facilitate disease transmission and NDD. Locally rare species may be less able to increase their equilibrium abundance without genetic boosts to defence via immigration of novel R gene alleles from a larger and more diverse regional population.

Polyhedrin sequence determines the tetrahedral shape of occlusion bodies in Thysanoplusia orichalcea single-nucleocapsid nucleopolyhedrovirus.

A nucleopolyhedrovirus (NPV) isolated from the looper Thysanoplusia orichalcea L. (Lepidoptera: Noctuidae) (ThorNPV) is occluded in a tetrahedral protein matrix. The ORF of the ThorNPV polyhedrin gene contains 738 nt which code for 246 amino acids of the putative polyhedrin protein with an estimated molecular mass of 28,778 Da. The promoter of this gene is similar in length to the promoter of Spodoptera frugiperda NPV (SfMNPV), with a 5 nt deletion before the start codon compared to those of other NPVs. When the polyhedrin gene of Autographa californica NPV (AcMNPV), whose occlusion bodies (OBs) are polyhedral, was replaced by the polyhedrin gene of ThorNPV, which produces tetrahedral OBs, tetrahedral polyhedra with properly occluded virions were produced. This work establishes the importance of the polyhedrin protein sequence in determining OB shape. Leucine at position 43 of ThorNPV polyhedrin was identified as responsible for the tetrahedral shape of ThorNPV OBs by PCR-based site-directed mutagenesis. Susceptibility to alkaline buffer of OBs formed by recombinant AcMNPV (RECAcV) carrying the polyhedrin gene of ThorNPV was slightly greater than that of native ThorNPV OBs. The LD50 of RECAcV for third-instar beet armyworm (Spodoptera exigua) was significantly lower than that of AcMNPV (253 and 31 OBs per larva, respectively).

Sex-specific and hormone-controlled expression of a vitellogenin-encoding gene in the gypsy moth.

Microvitellogenin and vitellogenin cDNA from Manduca sexta (tobacco hornworm) were tested for use as molecular probes to investigate the expression of genes coding for vitellogenins in Spodoptera frugiperda (fall armyworm) and Lymantria dispar (gypsy moth). Cross-hybridization was not observed between the M. sexta cDNAs and S. frugiperda DNA and mRNA. Vitellogenin cDNA from M. sexta did not hybridize to L. dispar DNA or mRNA. However, the 834 bp microvitellogenin cDNA from M. sexta hybridized to an approximately 850 bp transcript in L. dispar mRNA. A 2.5 kb cDNA clone, pz64, was isolated from late last instar larvae of female L. dispar by differential screening. This clone has 38% amino acid sequence (deduced) and 55% nucleic acid sequence similarities with the 3'-end of high molecular weight vitellogenin in Bombyx mori (silkworm). When used as a probe in northern analysis of L. dispar mRNA, this cDNA hybridized to a 5.3 kb transcript in female last instar larvae, pupae, and adults, but not to male last instar larvae and adults. This cDNA did not hybridize to mRNA from M. sexta or S. frugiperda. Expression of the 5.3 kb vitellogenin transcript hybridizing to the 2.5 kb cDNA clone was suppressed in 5-day-old last instar larvae of female L. dispar treated on day 2 with doses of the juvenile hormone analog, methoprene, greater than 10 nmol. Apparently, the high in vivo titer of juvenile hormone during the first 2 days of the last instar represses the transcription of vitellogenin mRNA.

Quantification of electroantennogram responses of the primary rhinaria ofAcyrthosiphon pisum (Harris) to C4-C 8 primary alcohols and aldehydes.

Electroantennograms (EAGs) of the distal and proximal primary rhinaria (DPR and PPR, respectively) were recorded from excised antennae of alate virginoparous pea aphids,Acyrthosiphon pisum (Harris) (Homoptera: Aphididae). Primary unsaturated alcohols and aldehydes with varying carbon length (C4-C8) were used as volatile stimuli. EAGs were recorded for a series of source concentrations from the DPR and PPR separately through the use of sectional electroantennography. A logistic equation was fitted to the source concentration-response data. Differences in relative EAG response of the DPR and PPR to the alcohols and aldehydes were analyzed by deriving five parameters from this logistic equation. These parameters relate to particular characteristics of sigmoid curves: the saturation (maximum) EAG response (R s ), the concentration for which the relative EAG response is ½R s (CR50), the stimulus response range (SR 0.9), the threshold concentration (CR1), and the EAG response area (A R ). Of these parameters, the EAG response area showed the largest separation between EAG responses of the DPR and PPR to the two homologous groups and between compounds with varying carbon chain lengths. The DPR was significantly more responsive to alcohols than to aldehydes, while the reverse was true for the PPR, indicating a basic difference between the two primary rhinaria. The highest overall responses were elicited by 1-hexanol, hexanal, and heptanal.

Male european corn borer,Ostrinia nubilalis (Hübner), antennal responses to analogs of its sex pheromone : Strain, electroantennogram, and behavior relationships.

Experiments were conducted to (1) determine whether the electroantennogram (EAG) can detect differences among the responses of antennae from males derived from the three strains ofOstrinia nubilalis (Hübner), and (2) characterize the EAG responses of each strain to isomeric forms of the natural pheromone, (E)- and (Z)-11-tetradecen-1-ol acetate (TDA), and analogs possessing differences in the terminal alkyl group, cyclopropyl (CPA), ortert-butyl (TBA).EAG responses differed among the strains in two ways: (1) Antennae fromZZ males always produced an EAG to (Z)-TDA with an extended duration of response. This "signature" EAG response was found to be unique to the antennal response ofZZ males to (Z)-TDA, thus providing a relatively easy method of distinguishing liveZZ males fromEE orZE males. Correlated with this longer EAG response was a longer disadaptation time, i.e., the EAG response ofZZ antennae disadapted more slowly (ca. 10 min) than the response ofEE antennae. (2) Strain differences in the relative EAG amplitudes to isomers and analogs were observed at the stimulus amounts eliciting the peak EAG amplitude as follows: TDA ≥ CPA > TBA forZZ males and both isomers; TDA > CPA ≥ TBA and CPA ≥ TDA > TBA forEE males and theE andZ isomers, respectively; CPA > TBA ≥ TDA forZE males and both isomers. Dose-response relationships were seen for all compounds if amplitude ("peak height") of the EAG was used as a measure of response. However, if width of the EAG at half the peak height ("peak width") was used, then only theZZ antennal response to (Z)-TDA resulted in a meaningful dose-response relationship. For all strains, the EAG amplitudes elicited by theZ isomers of any of the tested compounds were greater than those elicited by the correspondingE isomers. Therefore, correlations between the relative EAG and upwind flight responses were observed in theZZ (r = 0.86) andZE (r = 0.80) strains but were not correlated in theEE strain (r = 0.18). Temporal studies showed that adaptation, not postexcision deterioration, was responsible for the observed decreases in the EAG amplitude after repetitive stimulation or after stimulation with amounts in a descending order. Disa-daptation required at least 20 min for a moderate dose (10 µg for 1 sec). Developmental studies showed that antennae from 2-day-old adults had the greatest EAG response.

Regeneration and biosynthesis of dytiscid defensive agents (Coleoptera: Dytiscidae).

The defensive secretions of the dytiscid species,Agabus seriatus (Say) andAgabus obtusatus (Say), were qualitatively and quantitatively analyzed by high-pressure liquid chromatography. The intrinsic ability ofA. Seriatus andA. Obtusatus to regenerate their prothoracic gland defensive secretions under laboratory conditions was determined by analyzing the secretions every seventh day for five weeks. Both beetles regenerated ∼ 80% of their prothoracic gland components within two weeks.A. seriatus was injected with [4-(14)C]cholesterol and after a three-week regeneration period 7.5% of the(14)Clabel was found in the steroidal defensive secretion from the prothoracic glands.