COMPARISON OF SPRAY, POINT INOCULATION METHODS, AND FDK TO FACILITATE EARLY GENERATION SELECTION FOR FUSARIUM HEAD BLIGHT RESISTANCE IN WINTER WHEAT Tamburic-Ilincic, L. 1, 2*, Fedak, G. 3, and Schaafsma, A. W. 1, 2 1
Ridgetown College, University of Guelph, Ridgetown, Ontario, Canada Department of Plant Agriculture, University of Guelph, Guelph, Ontario, Canada 3 Eastern Cereal and Oilseed Research Centre, Ottawa, Ontario, Canada *Corresponding author: PH: 519/674-1611; E- mail:
[email protected] 2
INTRODUCION Fusarium head blight (FHB) caused by Fusarium graminearum (Schwabe) is an important disease of wheat (Triticum aestivum L.) in Canada, and worldwide (Sutton, 1982). Different types of wheat resistance to FHB have been reported, but in the breeding programs Type 1, or resistance to initial infection, and Type 2, or resistance to spread of symptoms within the head are used most often (Schroeder and Christensen, 1963, Mesterhazy, 1995). The most frequently used source of FHB Type 2 resistance worldwide is Sumai-3. Several authors reported that Type 1 and Type 2 resistance varied independently among cultivars (Schroeder and Christensen, 1963; McKendry et al., 2001; Desmeules et al., 2001), and that selection of genotypes with resistance to FHB depends on the inoculation technique used (Engle et al., 2001; Bockus et al., 2001). Shaner and Buechley (2001) proposed the expression of severity as the number of spikelets blighted, not the proportion of spikelets blighted, in order to avoid the influence of spike size on the degree of Type 2 resistance. Van Saford et al., (1999), and Hall et al., (2000) reported that some genotypes have a lower kernel infection rate then expected from their spikelets infection rate, and also the opposite. In order to increase the comprehensive resistant level to FHB, different resistance types to FHB should be pyramided into improved varieties (Xu et al., 2001). Even though spray and point inoculation methods are used most frequently as the ways to estimate wheat resistance or susceptibility to FHB, these two methods are not often directly compared using segregating populations with known type of FHB resistance. The objectives of this study are: -to directly compare FHB severity after spray and point inoculation method using a segregating population with type 2 FHB resistance, -to compare Type 1 and Type 2 resistance with Type 4, or resistance to kernel infection, -to compare expression of FHB severity as the number of spikelets infected with expression of FHB severity as the proportion of spikelets infected, in order to examine the influence of spike size on the degree of FHB resistance.
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MATERIALS AND METHODS The influence of different methods of inoculation on number or proportion of FHB infected spikelets, and percent of Fusarium damage kernels (FDK), were studied using an F3 population carrying Type 2 FHB resistance. The progeny (n=85) was derived from a cross of resistant (WEKO60DH3 - a Sumai 3 derivative) and susceptible (AC RON) parents. The segregating generations, and parents, were planted on October 20, 2000, in 2- m long single rows, spaced 17.8 cm apart, at Ridgetown, Ontario. In order to obtain uniform growth stage at the time of inoculation, individual heads, rather than whole plants or plots, were inoculated at 50 % of anthesis (Zadoks growth stage 60-69) (Zadoks, 1974). The heads were spray inoculated with 2 mL of the suspension sprayed onto individual heads, and point inoculated with 10 ? l of suspension injected into single florets. The suspension of macroconidia, including three isolates of F. graminearum, was produced in liquid shake culture using modified Bilay’s medium, and used at a concentration of 50,000 spores/mL. Between10 to 20 plants from each progeny and the parents were inoculated using both methods of inoculation. Clear plastic bags were placed over the inoculated heads, and left for 48 hr to maintain humidity. The plots were misted daily with an overhead mister that delivered about 7.5 mm of water each day. The plots were fertilized and maintained using provincial recommendations. The number of diseased spikelets and the total number of spikelets were recorded for each inoculated wheat head. The spikelet infection rate was calc ulated as the number of diseased spikelets, or percentage of diseased spikelets of the total number of spikelets. The average infection rate from each row was calculated. The inoculated heads were hand harvested separately. Heads from each row, with the same inoculation method, were threshed together using a single head thresher, retaining all light kernels. The number of healthy and fusarium damaged kernels were counted, and percentage of FDK was calculated for each line and parents, after both methods of inoculation. In order to avoid the influence of inoculation method on % of FDK, % of FDK after both methods of inoculation was also averaged for each line, and their parents. FDK were identified as shriveled kernels, with chalky, pink or white color. The F3 progeny was assigned to phenotypic classes on the basis of their position in the distribution of the proportion of FHB infected spikelets, and % of FDK. For the proportion of FHB infected spikelets the following classes were used: 1=0-2.5, 2=2.51-5, 3=5.01-7.5, 4=7.51-10, 5=10.01-12.5, 6=12.51-15, 7=15.01-17.5, 8=17.51-20, 9=20.01-22.5, 10=22.51-25, 11=25.01-27.5, 12=27.51-30, 13=30.01-32.5, 14=32.51-35, 15=35.01-37.5, 16>37.51. For % of FDK, there were the following classes: 1=0-2, 2=2.01-4, 3=4.01-6, 4=6.01-8, 5=8.01-10, 6=10.01-12, 7=12.01-14, 8=14.01-16, 9=16.01-18, 10=18.01-20, 11=20.01-22, 12>22.01. Data management and all statistical procedures were completed using SAS v. 6.0. (SAS Institute Inc, 2001). RESULTS AND DISCUSSION
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Transgressive segregants, with higher levels of resistance than the parents, were found using visual symptoms, and % FDK after both methods of inoculation (Fig. 1-2). Minimum, maximum, and mean values for % of infected spikelets after point inoculation were 3.7, 32.7, and 11.4, and lower than after spray inoculation where they were 4.8, 42.0, and 15.4, respectively. This result was expected because this population carring Type 2 resistance from Sumai-3. Overall correlation between % of diseased spikelets after spray and point inoculation was positive, but low (r=0.38, P
