A hypervariable intron of the STAYGREEN locus provides excellent discrimination among Pisum fulvum accessions and reveals evidence for a relatively recent hybridization event with Pisum sativum.

An analysis of 82 non-synonymous Pisum fulvum accessions for sequence variation in a fragment of the STAYGREEN (SGR) locus revealed 57 alleles, most of which differed in indel structure. Eight additional P. fulvum accessions, each supposedly synonymous with a different accession of the initial group, were also analyzed. In every case the paired synonymous accessions possessed the same SGR sequence but varied slightly for a 6-trait morphological phenotype, indicating that SGR sequence is a much more reliable indicator of accession identity than is a morphological characterization. SGR sequence analysis confirmed our previous finding that P. fulvum accessions separate into two allele groups. This division was not supported by results of previous studies that were based on sequences distributed across the entire genome, suggesting that the division may have been produced by selection at a nearby locus and that the SGR phylogeny may not be good indicator of overall relationships within the species. One P. fulvum accession, PI 595941 (=JI1796), displayed an SGR sequence outside the variation typical of the species. Instead, its allele resembled alleles limited to a set of Pisum sativum landraces from the Middle East, suggesting hybridization between ancestors of PI 595941 and some primitive form of domesticated P. sativum. With one exception from the extreme northwest corner of Israel, P. fulvum accessions collected north of latitude 35.5° N were fixed for alleles from group A. These northern accessions also displayed greatly reduced SGR sequence diversity compared to group A accessions collected from other regions, suggesting that the northern populations may represent recent extensions of the range of the species. Group B accessions were distributed from Lake Tiberias south and were generally sympatric with the southern group A accessions. Although group B accessions occupied a smaller area than group A, the SGR sequence diversity in this group (28 alleles in 33 accessions) exceeded that for group A.

Construction and characterization of two bacterial artificial chromosome libraries of pea (Pisum sativum L.) for the isolation of economically important genes.

Pea (Pisum sativum L.) has a genome of about 4 Gb that appears to share conserved synteny with model legumes having genomes of 0.2-0.4 Gb despite extensive intergenic expansion. Pea plant inventory (PI) accession 269818 has been used to introgress genetic diversity into the cultivated germplasm pool. The aim here was to develop pea bacterial artificial chromosome (BAC) libraries that would enable the isolation of genes involved in plant disease resistance or control of economically important traits. The BAC libraries encompassed about 3.2 haploid genome equivalents consisting of partially HindIII-digested DNA fragments with a mean size of 105 kb that were inserted in 1 of 2 vectors. The low-copy oriT-based T-DNA vector (pCLD04541) library contained 55 680 clones. The single-copy oriS-based vector (pIndigoBAC-5) library contained 65 280 clones. Colony hybridization of a universal chloroplast probe indicated that about 1% of clones in the libraries were of chloroplast origin. The presence of about 0.1% empty vectors was inferred by white/blue colony plate counts. The usefulness of the libraries was tested by 2 replicated methods. First, high-density filters were probed with low copy number sequences. Second, BAC plate-pool DNA was used successfully to PCR amplify 7 of 9 published pea resistance gene analogs (RGAs) and several other low copy number pea sequences. Individual BAC clones encoding specific sequences were identified. Therefore, the HindIII BAC libraries of pea, based on germplasm accession PI 269818, will be useful for the isolation of genes underlying disease resistance and other economically important traits.

First Report of Sclerotinia Stem Rot of Chickpea Caused by Sclerotinia sclerotiorum in North Dakota and Washington.

Chickpea (Cicer arietinum L.) is cultivated as a rotational crop in the cereal-based production system in the U.S. Pacific Northwest (PNW) and its production is expanding to other northern tier states. During July 2005, symptoms of Sclerotinia stem rot were observed on chickpea cv. Dwelley and Dylan in fields near Spangle, WA and Carrington, ND, respectively, with disease incidence of approximately ≤1% in affected areas at both locations. Symptoms included stem whitening, wilting, and stem breakage. Occasionally, white fluffy mycelium was observed; however, production of sclerotia on infected plants was rarely observed. Sclerotinia sclerotiorum was isolated from diseased stems collected from both states. The isolates produced a ring of sclerotia near the edge of potato dextrose agar (PDA) plates in 7 days and produced neither conidia nor other fruiting bodies in culture after 30 days. PCR amplification of the rDNA internal transcribed spacer region from two representative isolates and subsequent digestion with restriction enzymes, Mbo I and Taq I, produced identical banding patterns to previously identified isolates of S. sclerotiorum from pea from the PNW (2). Chickpea cvs. Dwelley and Spanish White (eight plants of each) were inoculated by fastening mycelial agar plugs from an actively growing colony on PDA onto the stems with Parafilm. Symptoms of stem whitening were observed as early as 2 days after inoculation, and the lesions extended upward and downward from the inoculation site. Wilting and stem breakage were also observed. Control inoculations of four plants of each cultivar with PDA plugs without mycelium produced no visible symptoms. S. sclerotiorum was consistently reisolated from inoculated plants but not from control plants. Chickpea had been grown in the PNW for more than 20 years without any reported incidence of Sclerotinia stem rot although the disease has been reported from Arizona (3) and Asian countries (1). This is likely because of the upright growth habit of the chickpea plant coupled with relatively dry conditions late in the growing season. Previous chickpea cultivars were very susceptible to Ascochyta blight, an early-season disease of chickpea in the PNW that reduced chickpea stands and canopy coverage. Current cultivars possess much improved resistance to Ascochyta blight, allowing greater vegetative growth to occur and creating microenvironmental conditions conducive to Sclerotinia stem rot. In North Dakota, where humid conditions prevail late in the growing season, symptoms of Sclerotinia stem rot had been observed in previous years but had not been documented because of a recent history of chickpea cultivation there. To our knowledge, this is the first report of confirmed Sclerotinia stem rot of chickpea in North Dakota and Washington. References: (1) G. J. Boland and R. Hall. Can. J. Plant Pathol. 16:93, 1994. (2) I. Jimenez-Hidalgo et al. Phytopathology (Abstr.) 94(suppl.):S47, 2004. (3) M. E. Matheron and M. Porchas. Plant Dis. 84:1250, 2000.

Microsatellite marker polymorphism and mapping in pea (Pisum sativum L.).

This paper aims at providing reliable and cost effective genotyping conditions, level of polymorphism in a range of genotypes and map position of newly developed microsatellite markers in order to promote broad application of these markers as a common set for genetic studies in pea. Optimal PCR conditions were determined for 340 microsatellite markers based on amplification in eight genotypes. Levels of polymorphism were determined for 309 of these markers. Compared to data obtained for other species, levels of polymorphism detected in a panel of eight genotypes were high with a mean number of 3.8 alleles per polymorphic locus and an average PIC value of 0.62, indicating that pea represents a rather polymorphic autogamous species. One of our main objectives was to locate a maximum number of microsatellite markers on the pea genetic map. Data obtained from three different crosses were used to build a composite genetic map of 1,430 cM (Haldane) comprising 239 microsatellite markers. These include 216 anonymous SSRs developed from enriched genomic libraries and 13 SSRs located in genes. The markers are quite evenly distributed throughout the seven linkage groups of the map, with 85% of intervals between the adjacent SSR markers being smaller than 10 cM. There was a good conservation of marker order and linkage group assignment across the three populations. In conclusion, we hope this report will promote wide application of these markers and will allow information obtained by different laboratories worldwide in diverse fields of pea genetics, such as QTL mapping studies and genetic resource surveys, to be easily aligned.

Discordance of physiological and biochemical response to smoking and to psychological stress.

Both smoking and psychological stress produce marked effects upon cardiovascular function, and several studies have demonstrated that in combination they produce additive or potentiating effects. More recently, it has been reported that individuals strongly reactive to psychological stress are also strongly reactive to nicotine. In an attempt to replicate and extend those findings, we examined reactivity to smoking and competitive mental arithmetic across several physiological and biochemical variables. Despite stable responding across mental arithmetic trials, we were unable to demonstrate significant correlations between reactivity to smoking and to a psychological stressor. We further observed that anxiety level, when low, was a poor predictor of desire to smoke and of withdrawal, whereas higher anxiety levels were more tightly linked to these measures. These findings have implications for the identification of individuals at risk of cardiovascular disease as well as for the design of smoking treatment and relapse prevention programs.

Simple phobia: evidence for heterogeneity.

Although simple phobia is a residual category in DSM-III, clinical experience suggests at least four subtypes of this group. To test the validity of the subtypes, the authors compared patients with one of four simple phobias subtypes (n: animal-insect = 25, blood-injury = 9, situational = 46, choking-vomit = 8). Significant sex differences were observed; all animal and insect phobics and seven of eight choking-vomit phobics were female, while the other two groups showed approximately equal numbers of males and females. Mean age of onset was significantly older for situational phobics than animal-insect or blood-injury phobics; choking-vomit probands were intermediate. Frequency of situational phobias differed significantly among relatives of the four proband groups, with highest frequency being found among situational probands. Thus, these clinical and epidemiological variables support the separation of simple phobia into at least these four diagnostic groups.

Menstrual fluctuation in the symptoms of panic anxiety.

Ten women with DSM-III-defined panic attacks (five with and five without agoraphobia) had symptom severity rated daily, weekly, and retrospectively through one full menstrual cycle. Substantial fluctuations in retrospective ratings of severity were observed, with the premenstrual week being rated as most severe. Daily and weekly ratings showed much smaller fluctuations in the predicted direction. Possible reasons for this outcome are considered.

Circadian symptom fluctuations in people with anxiety disorders.

Circadian rhythm abnormalities have been demonstrated in people with depression, including a tendency toward maximal symptom severity in the morning. Although a few studies have suggested that symptoms in people with anxiety are worse later in the day, no detailed study of this observation has been reported. In 86 patients with anxiety disorders (63 with panic disorder or agoraphobia with panic attacks), anxiety symptoms tended to be more severe in the afternoon or evening than in the morning, with no abnormalities of heart rate or oral temperature. This is the first systematic demonstration of a circadian fluctuation of mood in any disorder other than depression.