Genotyping Common Bean for the Potyvirus Resistance Alleles I and bc-1(2) with a Multiplex Real-Time Polymerase Chain Reaction Assay

ABSTRACT A multiplex real-time polymerase chain reaction (PCR) assay was developed to simultaneously genotype plants for the I and bc-1(2) alleles, which condition resistance in beans to Bean common mosaic virus and Bean common mosaic necrosis virus. A segregating F(2) population was derived from the cross between pinto bean breeding line P94207-189A (bc-1 bc-1 I I) and Olathe (bc-12 bc-1(2) i i). Real-time PCR assays were developed that were specific for each allele, and a multiplex PCR reaction could unambiguously assign F(2) plants to one of nine genotypes. Remnant F(1) plants were used as a comparative reference sample. PCR results among this sample fit a normal distribution for both real-time PCR assays, and 99% probability distributions were determined for heterozygotes. F(2) plants were genotyped based on results relative to the probability distributions for heterozygotes. F(2) plants also were genotyped for the I and bc-1(2) alleles by performing F(3) family progeny tests for virus resistance. Agreement between the two methods was 100% (198/198) for the bc-1(2) allele, and 92.4% (183/198) for the I allele. Erroneous genotyping was due to recombination between the amplicon and the I allele. Realtime PCR assays provide a robust method for genotyping seedlings and, in some cases, may eliminate the need for progeny testing.     

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Response of Ugandan common bean varieties to Pseudocercospora griseola and Angular leaf spot disease development in varietal mixtures

Angular leaf spot (ALS) is one of the most economically important bean diseases in Africa. One promising control option for the disease is the use of mixtures of resistant and susceptible varieties. This research evaluated (1) the reaction of farmer preferred bean varieties to ALS in the screenhouse and on-station and (2) the effect of different spatial arrangements of resistant and susceptible bean varieties on ALS disease development. For the latter, five mixture combinations and two controls were laid out in a Randomized Complete Block Design having three replicates. Analysis of variance and least significant differences (LSDs) were used to compare disease levels in both studies above. Varietal screening showed significant differences in varietal reaction to ALS. Screenhouse disease scores ranged between 0 and 5, compared to 0.7–3.9 in the field. For the varietal mixture trial, the lowest disease levels and the highest mixture efficiencies were observed for the combination of equal proportions of the susceptible and resistant varieties randomly mixed (even mixture) prior to planting. We conclude that even mixtures reduce the amount of ALS disease that develops in the season.     

Using 3D animations to teach intracellular signal transduction mechanisms: taking the arrows out of cells

Traditional methods of teaching intracellular biological processes and pathways use figures or flowcharts with the names of molecules linked with arrows. Many veterinary students, presented with such material, simply memorize the names or chemical structures of the molecules and are then likely to forget the material once the examination is completed. To address this problem, the authors designed, created, and field-tested new teaching media that incorporate realistic three-dimensional (3D) animations depicting the dynamic changes that occur in intracellular molecules during cellular activation. Testing found that veterinary students taught using traditional teaching media (e.g., lectures, handouts, textbooks) are proficient in memorizing the names and order of intracellular molecules but unable to appreciate the interactions between these elements or their spatial relationships within cells. In contrast, more than 90% of veterinary students taught using 3D animations not only recall the facts about the intracellular elements but also develop accurate mental images of the interactions among these molecules and their spatial relationships. These findings strongly suggest that the comprehension of complex biological processes by veterinary students can be enhanced by the use of dynamic 3D depictions of these processes in the classroom.     

Inheritance of leaf angle in Triticum aestivum L.

Summary A 6×6 diallel was prepared to study the inheritance of leaf angle in T. aestivum L. Genetic analysis in terms of diallel cross parameters and graphic analysis indicated the control of additive gene effects in the expression of this character. The results of F₁ analysis were supported by the analysis of F₂ data.     

Seedling emergence,coleoptile length,and plant height relationships in crosses of dwarf and standard-height wheats

Summary Seedling emergence was closely correlated with coleoptile length and plant height among parents, F₂ and F₃, populations of crosses involving dwarf wheats Olesen Dwarf (CI 14497), Norin 10 derivative D6301, Tom Thumb derivative D6899, and the standard-height varieties Ramona 50 and Nainari 60. Genetic mechanisms that governed plant height also influenced coleoptile length, but the relative effects of genes showing dominant or epistatic effects appeared to be different. With respect to the two parents involved in each of 15 crosses, mean F₂ coleoptile lengths were consistently closer to the low parent value than were corresponding mean F₂ plant heights. A slight curvilinear relationship was also found between coleoptile length and plant height of F₃ lines. The results suggest that selection of semidwarf wheats with long coleoptiles and improved emergence properties from crosses involving the dwarf wheats of this study would be unlikely.     

Analysis of rice blast resistance gene <Emphasis Type="Italic">Pi</Emphasis>-<Emphasis Type="Italic">z</Emphasis> in rice germplasm using pathogenicity assays and DNA markers

The Pi-z gene in rice confers resistance to a wide range of races of the rice blast fungus, Magnaporthe oryzae. The objective of this study was to characterize Pi-z in 111 rice germplasm accessions using DNA markers and pathogenicity assays. The existence of Pi-z in rice germplasm was detected by using four simple sequence repeat (SSR) markers (RM527, AP4791, AP5659-1, AP5659-5) closely linked to Pi-z, and was verified using pathogenicity assays with an avirulent strain (IE1k) and two virulent races (IB33 and IB49). Among 111 germplasm accessions evaluated, 73 were found to contain the Pi-z gene using both SSR markers and pathogenicity assays. The remaining 38 germplasm accessions were found to be inconsistent in their responses to the blast races IB33, IEIk and IB49 with expected SSR marker alleles, suggesting the presence of unexpected SSR alleles and additional R gene(s). These characterized germplasm can be used for genetic studies and marker-assisted breeding for improving blast resistance in rice.     

Assessing the Impacts of Time-to-Detection Distribution Assumptions on Detection Probability Estimation

Abundance estimates from animal point-count surveys require accurate estimates of detection probabilities. The standard model for estimating detection from removal-sampled point-count surveys assumes that organisms at a survey site are detected at a constant rate; however, this assumption can often lead to biased estimates. We consider a class of N-mixture models that allows for detection heterogeneity over time through a flexibly defined time-to-detection distribution (TTDD) and allows for fixed and random effects for both abundance and detection. Our model is thus a combination of survival time-to-event analysis with unknown-N, unknown-p abundance estimation. We specifically explore two-parameter families of TTDDs, e.g., gamma, that can additionally include a mixture component to model increased probability of detection in the initial observation period. Based on simulation analyses, we find that modeling a TTDD by using a two-parameter family is necessary when data have a chance of arising from a distribution of this nature. In addition, models with a mixture component can outperform non-mixture models even when the truth is non-mixture. Finally, we analyze an Ovenbird data set from the Chippewa National Forest using mixed effect models for both abundance and detection. We demonstrate that the effects of explanatory variables on abundance and detection are consistent across mixture TTDDs but that flexible TTDDs result in lower estimated probabilities of detection and therefore higher estimates of abundance.Supplementary materials accompanying this paper appear on-line.     

Current and historical inputs of mercury to high-latitude lakes in Canada and to Hudson Bay

Sediment cores collected from several lakes in northern Canada have been analyzed for mercury and several other chemical contaminants. Sites ranged from the Experimental Lakes Area of northwestern Ontario, north to Cornwallis Island, and west to the southern Yukon. Cores were sliced at sites of collection and individual slices were freeze dried and analyzed for Pb-210 and Cs-137 to estimate average time intervals of deposition. The earliest date estimated by Pb-210 was about 1850, and mercury concentrations in some lakes were clearly increasing before then, assuming no vertical movements of mercury within the sediments. Extrapolation of dates downward to deeper slices, assuming a constant sedimentation rate, indicated that in some lakes mercury inputs increased slowly even in the 1500's, more rapidly after 1750, and more rapidly yet over the current century. These increases are interpreted as increased fluxes of mercury to the lakes as a result of long-range transport of atmospheric mercury, since there are no local industrial sources of mercury. Slices taken near the bottom of a core are taken to estimate the geological component while elevations in excess of that in surface slices are taken to represent contamination from fallout. This partitioning suggests that sediments in the eastern Northwest Territories are dominated by pollution, while those from the western Northwest Territories are influenced more by their geological settings. Two cores from Hudson Bay suggest that mercury is increasing there too, but has not yet exceeded geological sources. Mercury shows little or no tendency to decline in the most recent slices; indicating that inputs of mercury remain at or near their historical maxima. Given relatively high and continuing inputs of mercury to northern lakes it seems likely that some portion of that mercury may find its way into the food chain, hence the long-term prospect is for increasing levels of mercury in northern fish.