Association of AFLP markers with downy mildew resistance in autotetraploid alfalfa

The amplified fragment length polymorphism (AFLP) assay is an efficient method for the identification of molecular markers useful in the improvement of numerous crop species. The identification of AFLP markers linked to disease resistance genes has been shown in segregating populations from crosses of inbred lines. The development of inbred lines in alfalfa is not possible, but existing breeding programs have produced populations selected for resistance to a single pest. Two such populations, UC-123 and UC-143, differing only in selection for resistance to downy mildew (Peronospora trifoliorum de Bary) isolate I-8, were used in this study. Thirty-six resistant plants from UC-143, and 36 susceptible plants from UC-123 were screened for DNA polymorphisms using fourteen AFLP primer combinations. Four AFLP fragment markers, ACACTC₂₀₈, ACACTC₁₅₀, ACACAT₂₁₆ and ACACTC₄₈₆, were found to be significantly associated with disease susceptibility or resistance. Resistant and susceptible plants were crossed in a diallel scheme and the progeny were screened for resistance to P. trifoliorum isolate I8. Two of the AFLP markers, ACACTC₂₀₈ and ACACTC₄₈₆ were significantly associated with resistance in the F₁ and S₁ progeny. The utilization of two populations, comprised of 36 resistant and 36 susceptible plants, for the identification of DNA fragments associated with disease resistance proved successful. Seventy-two plants is a very manageable number and provides a starting point for further refinement of marker-trait associations.     

Comparing two methods for aging trees with suppressed,diffuse-porous rings (Betula pubescens ssp. czerepanovii)

The annual growth rings of diffuse porous species such as mountain birch are often difficult to distinguish when samples are collected from trees that grow at treeline or in other harsh environments. In this study we document the differences in seedling and sapling ring counts obtained from two methods of analysis: a traditional analysis based on reflected light and low-power microscopy and one based on transmitted light with higher power magnification that uses thin-sections of the samples. Rings are easier to resolve using the more labor-intensive transmitted light method. Small rings are often missed when using the reflected light method, resulting in an underestimation of tree age. The dates estimated by the standard method agreed with those determined using the thin-sectioning method in 9.6% of the cases. Most commonly, the standard method gave a younger age than did thin-sectioning (72.4% of the trees). In only 18.03% of the cases did the standard method result in a greater age than did thin-sectioning. The reflected light method produced age estimations that were on average 1.37 years younger than those determined using the transmitted light method. The difference between the two methods was positively correlated with age and negatively correlated with mean ring-width. Age-class histograms based on the two methods show little difference at coarser aggregation levels (decades and pentads), but annualized age-class histograms have less agreement between the two methods. Therefore, we suggest using the more labor-intensive thin-sectioning method when annualized age counts are necessary in suppressed seedlings and saplings, for example, comparing tree establishment with annual climate conditions at treeline.