Developmental changes in the germinability, desiccation tolerance, hardseededness, and longevity of individual seeds of Trifolium ambiguum

Background and Aims: Using two parental clones of outcrossing Trifolium ambiguum as a potential model system, we examined how during seed development the maternal parent, number of seeds per pod, seed position within the pod, and pod position within the inflorescence influenced individual seed fresh weight, dry weight, water content, germinability, desiccation tolerance, hardseededness, and subsequent longevity of individual seeds. Methods: Near simultaneous, manual reciprocal crosses were carried out between clonal lines for two experiments. Infructescences were harvested at intervals during seed development. Each individual seed was weighed and then used to determine dry weight or one of the physiological behaviour traits. Key Results: Whilst population mass maturity was reached at 33–36 days after pollination (DAP), seed-to-seed variation in maximum seed dry weight, when it was achieved, and when maturation drying commenced, was considerable. Individual seeds acquired germinability between 14 and 44 DAP, desiccation tolerance between 30 and 40 DAP, and the capability to become hardseeded between 30 and 47 DAP. The time for viability to fall to 50 % (p50) at 60 % relative humidity and 45 °C increased between 36 and 56 DAP, when the seed coats of most individuals had become dark orange, but declined thereafter. Individual seed f. wt at harvest did not correlate with air-dry storage survival period. Analysing survival data for cohorts of seeds reduced the standard deviation of the normal distribution of seed deaths in time, but no sub-population showed complete uniformity of survival period. Conclusions: Variation in individual seed behaviours within a developing population is inherent and inevitable. In this outbreeder, there is significant variation in seed longevity which appears dependent on embryo genotype with little effect of maternal genotype or architectural factors.

Relationship between incidence and severity of cashew gummosis in semiarid north-eastern Brazil

The incidence-severity relationship for cashew gummosis, caused by Lasiodiplodia theobromae, was studied to determine the feasibility of using disease incidence to estimate indirectly disease severity in order to establish the potential damage caused by this disease in semiarid north-eastern Brazil. Epidemics were monitored in two cashew orchards, from 1995 to 1998 in an experimental field composed of 28 dwarf clones, and from 2000 to 2002 in a commercial orchard of a single clone. The two sites were located 10 km from each other. Logarithmic transformation achieved the best linear adjustment of incidence and severity data as determined by coefficients of determination for place, age and pooled data. A very high correlation between incidence and severity was found in both fields, with different disease pressures, different cashew genotypes, different ages and at several epidemic stages. Thus, the easily assessed gummosis incidence could be used to estimate gummosis severity levels.

Flowering intensity in white yam (Dioscorea rotundata)

White or Guinea yam (Dioscorea rotundata), grown for its underground tubers, is an important food in West Africa. Progress in yam breeding is constrained by variable flowering behaviour, making hybridization difficult. Yam clones may be dioecious, monoecious or hermaphrodite with variable sex ratios. The proportion of plants that flower and the flowering intensity also vary with season and location. The objective of the present work was to investigate whether variation in flowering behaviour was related to factors determining rate of development (photoperiod and temperature through sowing date, location and year) or growth (cumulative solar radiation and temperature). Sex ratios, the proportion of plants that had flower buds and open flowers, and the number of flowers or spikes was recorded in one male (TDr 131) and one female (TDr 99-9) clone of white yam grown in the field in Nigeria at three locations and at different sowing dates. Clone TDr 131 was uniformly male flowering, while clone TDr 99-9 exhibited a number of sex types with gynoecious, monoecious and trimonoecious plants observed. The proportion of flowering plants was low in both clones, averaging 0.34 in clone TDr 131 and 0.13 in clone TDr 99-9. Day of vine emergence had a significant and contrasting effect on the proportion of flowering plants and on flowering intensity in the two clones. In clone TDr 131, the proportion of flowering plants and flowering intensity declined with later vine emergence at all locations (r=0.43-0.53, P<0.05), whereas in clone TDr 99-9 the proportion of flowering plants increased with later emergence (r=0.46, P<0.01). In clone TDr 131, this response was strongly associated with warmer temperatures (r=0.49-0.50; P<0.05) and greater cumulative radiation (r=0.85-0.93; P<0.001) between vine emergence and flowering, rather than photoperiod at vine emergence. This suggests that flowering behaviour in the male clone TDr 131 is strongly influenced by factors that affect growth rather than development. Clone TDr 99-9, on the other hand, exhibited no clear relations between flowering and growth or developmental factors, though the proportion of flowering plants and flowering intensity was greatest at planting dates close to the longest day and at temperatures of 25-26 degrees C. This might suggest that flowering behaviour in clone TDr 99-9 is controlled by photothermal responses.

Duration from vine emergence to flowering suggests a long-day or rate of change of photoperiod response in white yam (Dioscorea rotundata Poir.)

The objective of this study was to quantify the effect of photoperiod on the duration from vine (shoot) emergence to flowering in white or Guinea yam (Dioscorea rotundata). The duration from vine emergence to flowering in two clonal varieties of yam (TDr 131 and TDr 99-9) was recorded at 10 different sowing dates/locations in Nigeria. Durations to flowering varied from 40 to > 88 days. Mean daily temperature and photoperiod between vine emergence and flowering varied from 25 to 27 degrees C and 13.1 to 13.4 h day(-1), respectively. Both clones had similar responses to temperature, with base and optimum temperatures of 12 and 25-27 degrees C, respectively. Thermal durations to flowering were strongly related (r(2) > 0.75-0.83) to absolute photoperiod (h) at vine emergence as well as to rate of change of photoperiod (s day(-1)) at vine emergence. The response to absolute photoperiod suggests that white yams are quantitative LDPs, flowering sooner in long than short days. Yams also flowered earlier when the rate of change of photoperiod was positive but small, or was negative. It is suggested that yams may use a combination of photoperiod and rate of change in order to fine tune flowering time. (c) 2006 Elsevier B.V. All rights reserved.

Effect of harvest date on the dormancy period of yam (Dioscorea rotundata)

Tuber dormancy enables yams to survive in the ground during the dry season and post-harvest storage. Three clones of Dioscorea rotundata were harvested after five intervals and then stored in a cooler (20.6°C) or at ambient temperature (27.8°C). The time from harvest to sprouting was shorter as harvest was delayed. The period from sowing to sprouting for each clone was similar for tubers harvested from 140 days after planting, but tubers harvested earlier took longer to sprout. The cooler temperature delayed sprouting. Tubers of two clones sprouted after only 70 days of crop growth. If the dormancy period of these young tubers can be broken, the generation time of yam crop improvement programmes could be considerably reduced.

The stimulation of mitogenic signaling pathways by N-POMC peptides

The N-terminal fragment of pro-opiomelancortin (POMC) has been shown previously to act as an adrenal mitogen. However, little is known about the molecular mechanisms by which mitogenesis is stimulated, although it has been shown that N-POMC1-28 Stimulates the ERK pathway in human H295R cells. We have investigated signaling stimulated by N-POMC1-28 and N-POMC1-49 in the mouse Y1 cell line and found that both peptides stimulate ERK phosphorylation with maximal stimulation being achieved within 5 min. Similar results were observed for both MEK and c-Raf phosphorylation, although N-POMC1-49 stimulated the phosphorylation of Akt more robustly than N-POMC1-28. We also investigated the expression of tyrosine kinase receptors in adrenal cells. PCR utilizing degenerate primers was performed on cDNA from both Y1 cells and rat adrenal tissue. Sequencing of 114 clones from each cDNA population revealed the expression of a number of receptors, several of which have not been described previously in the adrenal. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

In vivo expression technology (IVET) selection of genes of Rhizobium leguminosarum biovar viciae A34 expressed in the rhizosphere

IVET was used to identify genes that are specifically expressed in the rhizosphere of the pea-nodulating bacterium Rhizobium leguminosarum A34. A library of R. leguminosarum A34 cloned in the integration vector pIE1, with inserts upstream of a promoter-less purN:gfp:gusA, was conjugated into purN host RU2249 and recombined into the genome. After removal of colonies that expressed the reporter genes of the vector under laboratory conditions, the library was inoculated into a nonsterile pea rhizosphere. The key result is that 29 rhizosphere-induced loci were identified. Sequence analysis of these clones showed that a wide variety of R. leguminosarum A34 genes are expressed specifically in the rhizosphere including those encoding proteins involved in environmental sensing, control of gene expression, metabolic reactions and membrane transport. These genes are likely to be important for survival and colonization of the pea rhizosphere.

Display of a Maize cDNA library on baculovirus infected insect cells

Background: Maize is a good model system for cereal crop genetics and development because of its rich genetic heritage and well-characterized morphology. The sequencing of its genome is well advanced, and new technologies for efficient proteomic analysis are needed. Baculovirus expression systems have been used for the last twenty years to express in insect cells a wide variety of eukaryotic proteins that require complex folding or extensive posttranslational modification. More recently, baculovirus display technologies based on the expression of foreign sequences on the surface of Autographa californica ( AcMNPV) have been developed. We investigated the potential of a display methodology for a cDNA library of maize young seedlings. Results: We constructed a full-length cDNA library of young maize etiolated seedlings in the transfer vector pAcTMVSVG. The library contained a total of 2.5 x 10(5) independent clones. Expression of two known maize proteins, calreticulin and auxin binding protein (ABPI), was shown by western blot analysis of protein extracts from insect cells infected with the cDNA library. Display of the two proteins in infected insect cells was shown by selective biopanning using magnetic cell sorting and demonstrated proof of concept that the baculovirus maize cDNA display library could be used to identify and isolate proteins. Conclusion: The maize cDNA library constructed in this study relies on the novel technology of baculovirus display and is unique in currently published cDNA libraries. Produced to demonstrate proof of principle, it opens the way for the development of a eukaryotic in vivo display tool which would be ideally suited for rapid screening of the maize proteome for binding partners, such as proteins involved in hormone regulation or defence.

The severity of malarial anaemia in Plasmodium chabaudi infections of BALB/c mice is determined independently of the number of circulating parasites

Background: Severe malarial anaemia is a major complication of malaria infection and is multifactorial resulting from loss of circulating red blood cells (RBCs) from parasite replication, as well as immune-mediated mechanisms. An understanding of the causes of severe malarial anaemia is necessary to develop and implement new therapeutic strategies to tackle this syndrome of malaria infection. Methods: Using analysis of variance, this work investigated whether parasite-destruction of RBCs always accounts for the severity of malarial anaemia during infections of the rodent malaria model Plasmodium chabaudi in mice of a BALB/c background. Differences in anaemia between two different clones of P. chabaudi were also examined. Results: Circulating parasite numbers were not correlated with the severity of anaemia in either BALB/c mice or under more severe conditions of anaemia in BALB/c RAG2 deficient mice (lacking T and B cells). Mice infected with P. chabaudi clone CB suffered more severe anaemia than mice infected with clone AS, but this was not correlated with the number of parasites in the circulation. Instead, the peak percentage of parasitized RBCs was higher in CB-infected animals than in AS-infected animals, and was correlated with the severity of anaemia, suggesting that the availability of uninfected RBCs was impaired in CB-infected animals. Conclusion: This work shows that parasite numbers are a more relevant measure of parasite levels in P. chabaudi infection than % parasitaemia, a measure that does not take anaemia into account. The lack of correlation between parasite numbers and the drop in circulating RBCs in this experimental model of malaria support a role for the host response in the impairment or destruction of uninfected RBC in P. chabaudi infections, and thus development of acute anaemia in this malaria model.

Clonal variation in acetylcholinesterase biomarkers and life history traits following OP exposure in Daphnia magna

Two clones of Daphnia magna (Standard and Ruth) were exposed for 7 days to sub-lethal concentrations of acephate (5.0 and 10.0 mg/L). Survivorship, individual growth, reproduction and the population growth rate (lambda) were evaluated over three weeks. Acetylcholinesterase (AChE) activity was measured on days 2, 7 and 21. Acephate exposure inhibited AChE activity but had no direct effect on life history (LH) traits. There was also no effect of clone on AChE activity, LH and lambda. However, a significant interaction between clone and acephate concentration was found on both fecundity and AChE inhibition at 48 h was associated with a decrease in lambda the Standard clone and an increase in lambda in clone Ruth. Therefore, our findings show that genotypic variation will influence the link between AChE activity and toxic effects at higher levels of biological organisation in D. magna. (c) 2007 Elsevier Inc. All rights reserved.

Duration from vine emergence to flowering suggests a long-day or rate of change of photoperiod response in white yam (Dioscorea rotundata Poir.)

The objective of this study was to quantify the effect of photoperiod on the duration from vine (shoot) emergence to flowering in white or Guinea yam (Dioscorea rotundata). The duration from vine emergence to flowering in two clonal varieties of yam (TDr 131 and TDr 99-9) was recorded at 10 different sowing dates/locations in Nigeria. Durations to flowering varied from 40 to > 88 days. Mean daily temperature and photoperiod between vine emergence and flowering varied from 25 to 27 degrees C and 13.1 to 13.4 h day(-1), respectively. Both clones had similar responses to temperature, with base and optimum temperatures of 12 and 25-27 degrees C, respectively. Thermal durations to flowering were strongly related (r(2) > 0.75-0.83) to absolute photoperiod (h) at vine emergence as well as to rate of change of photoperiod (s day(-1)) at vine emergence. The response to absolute photoperiod suggests that white yams are quantitative LDPs, flowering sooner in long than short days. Yams also flowered earlier when the rate of change of photoperiod was positive but small, or was negative. It is suggested that yams may use a combination of photoperiod and rate of change in order to fine tune flowering time. (c) 2006 Elsevier B.V. All rights reserved.

Aphidius ervi preferentially attacks the green morph of the pea aphid, Acyrthosiphon pisum

The pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae) is found in red and green color morphs. Previous work has suggested that the aphidiine parasitoid Aphidius ervi Haliday preferentially attacks green pea aphids in the field. It is not clear whether these results reflect a real preference, or some unknown clonal difference, such as in immunity, between the aphids used in the previous studies. We used three susceptibility-matched pairs of red and green morph pea aphid clones to test for preferences. In a no-choice situation, the parasitoids attacked equal proportions of each color morph. When provided with a choice, A. ervi was significantly more likely to oviposit into colonies formed from green morphs when the neighboring colony was formed from red morph aphids. In contrast, red morphs were less likely to be attacked when their neighboring colony was of the green morph. By preferentially attacking green colonies, A. ervi may reduce the likelihood of intraguild predation, as it is suggested that visually foraging predators preferentially attack red aphid colonies. Furthermore, if this host choice behavior is replicated in the field, we speculate that color morphs of the pea aphid may interact indirectly through their shared natural enemies, leading to intraspecific apparent competition.

A qualitative host-pathogen interaction in the Theobroma cacao-Moniliophthora perniciosa pathosystem

The aim of this study was to test whether resistance of clones of Theobroma cacao ( cocoa) varied between isolates of Moniliophthora (formerly Crinipellis) perniciosa, the cause of witches' broom disease. Developing buds of vegetatively propagated T. cacao grown in greenhouses in the UK were inoculated with 16 000 spores of M. perniciosa per meristem in water, under conditions where water condensed on the inoculated shoot for at least 12 h after inoculation. The proportion of successful inoculations varied between clones and was inversely correlated with time to symptom production or broom formation. A specific interaction was demonstrated among three single-spore isolates of M. perniciosa and the clone Scavina 6 (SCA 6) and a variety of susceptible clones. Isolates Castenhal-I and APC3 were equally likely to infect SCA 6 and the other clones, but isolate Gran Couva A9 never infected SCA 6, although it was as virulent on the other clones. The interaction was maintained when the wetness period was extended to 70 h. Offspring of SCA 6 x Amelonado matings were all susceptible to both Castenhal-I and GC-A5, with no evidence of greater variability in susceptibility to GC-A5 than Castanhal-I. This suggests recessive inheritance of a single homozygous factor conferring resistance to GC-A5, from SCA 6. The progenies were slightly more susceptible to Castanhal-I than GC-A5. The implications for managing the disease are discussed.

Burkholderia Hep_Hag autotransporter (BuHA) proteins elicit a strong antibody response during experimental glanders but not human melioidosis

BACKGROUND: The bacterial biothreat agents Burkholderia mallei and Burkholderia pseudomallei are the cause of glanders and melioidosis, respectively. Genomic and epidemiological studies have shown that B. mallei is a recently emerged, host restricted clone of B. pseudomallei. RESULTS: Using bacteriophage-mediated immunoscreening we identified genes expressed in vivo during experimental equine glanders infection. A family of immunodominant antigens were identified that share protein domain architectures with hemagglutinins and invasins. These have been designated Burkholderia Hep_Hag autotransporter (BuHA) proteins. A total of 110/207 positive clones (53%) of a B. mallei expression library screened with sera from two infected horses belonged to this family. This contrasted with 6/189 positive clones (3%) of a B. pseudomallei expression library screened with serum from 21 patients with culture-proven melioidosis. CONCLUSION: Members of the BuHA proteins are found in other Gram-negative bacteria and have been shown to have important roles related to virulence. Compared with other bacterial species, the genomes of both B. mallei and B. pseudomallei contain a relative abundance of this family of proteins. The domain structures of these proteins suggest that they function as multimeric surface proteins that modulate interactions of the cell with the host and environment. Their effect on the cellular immune response to B. mallei and their potential as diagnostics for glanders requires further study.

Intraspecific heritable variation in life-history traits can alter the outcome of interspecific competition among insect herbivores

Competition is one of the most important biotic factors determining the structure of ecological communities. In this study, we show that there is variation in competitive ability between two clones of the pea aphid, Acyrthosiphon pisum, both of which out-compete a clone of the vetch aphid, Megoura viciae, in the laboratory. We tested whether this variation in competitive ability would alter the outcome of interspecific competition in the field. White one pea aphid clone followed the pattern set in the laboratory, out-competing the Megoura viciae clone, another showed the reverse effect with Megoura viciae dominating. These differences appear to be the result of variation in early population growth rate between the pea aphid clones, rather than predation, although predation did lead to the eventual extinction of colonies. We also questioned whether intra- and interspecific differences in predator escape behaviour could affect the outcome of competition in the field. All three clones responded similarly to the presence of foraging hoverfly larvae (Episyrphus balteatus), but the Megoura viciae clone dropped from the plant significantly less often in response to the presence of a foraging two-spot ladybird (Adalia bipunctata). This work provides evidence that intraspecific variation in competitive ability can alter the outcome of interspecific competitive interactions in nature and suggests that species-specific behavioural. traits may have the potential to modify the outcome of these interactions. (c) 2005 Gesellschaft fur Okologie. Published by Elsevier GmbH. All rights reserved.