Effects of Host Resistance on Colonisation of Brassica napus (Oilseed Rape) by Leptosphaeria maculans and Leptosphaeria biglobosa (Phoma Stem Canker)
Mohamed Sidique, Siti Nordahliawate
Leptosphaeria maculans and L. biglobosa co-infect winter oilseed rape plants to cause phoma stem canker disease. The sexual spores of both species are produced in pseudothecia on infected winter oilseed rape stem debris after harvest and this is the most important source of inoculum for infection of newly-emerged plants in autumn. Field experiments investigated the effects of host resistance on proportions of pathogens Leptosphaeria maculans and L. biglobosa in most leaf and stem tissues during 2011/2012, 2012/2013 and 2013/2014 growing seasons and on the pseudothecial development on crop debris on nine winter oilseed rape cultivars; cvs Adriana (Rlm4 + quantitative resistance (QR)), Bilbao (Rlm4), Capitol (Rlm1), Drakkar (no R gene against L. maculans), DK Cabernet (Rlm1 + (QR), Es-Astrid (QR), Excel (Rlm7), Roxet (Rlm7) and NK Grandia (QR). Cultivars with a combination of R-gene resistance and QR [Adriana (Rlm4 + QR), DK Cabernet (Rlm1 + QR)] or cultivars with only QR [(Es-Astrid and NK Grandia)] had more numbers of L. maculans leaf spots than other cultivars in autumn but less stem canker damage. There was greater number of L. biglobosa leaf spots on leaves of cvs Roxet and Excel with resistance gene Rlm7 than those of other cultivars and later more L. biglobosa DNA was detected in their stems than in those of other cultivars. In all cultivars in the three growing seasons, there was a greater amount of L. biglobosa DNA than L. maculans DNA in basal stem canker and upper stem lesions. The cv. Drakkar (no R gene against L. maculans) was susceptible in all three growing seasons, with a great number of L. maculans and L. biglobosa leaf spots and severe stem cankers. There were four cultivars (Adriana, Bilbao, Drakkar and NK Grandia) selected for the study of pseudothecial development under natural conditions with different times of exposure and in controlled environment conditions (20oC, continuous wetness). The fastest development was on the susceptible cv. Drakkar (no R gene against L. maculans), followed by Bilbao (Rlm4), Adriana (Rlm4 + QR) and NK Grandia (QR) for stem base cankers and upper stem lesions in controlled conditions. Results for pseudothecial development on stems of the nine winter oilseed rape cultivars that were exposed in natural conditions at Bayfordbury support the controlled environment results, with pseudothecia on stems of cultivars with a combination R-gene and QR consistently maturing later than those on other cultivars, regardless of the weather conditions in three growing seasons. Ascospores produced in pseudothecia are the primary inoculum that initiate phoma stem canker epidemics in autumn. Ascospore release was later in autumn 2011 than in autumn/winter 2012/2013 or 2013/2014 because of dry weather. The pattern of ascospore release had a peak, or maximum in autumn/winter 2011/2012 (4958 spores/m-3 on 22 Jan 2012) and several maxima in autumn/winter 2012/2013 (1307 spores/m-3 on 5 Nov 2012, 1291 spores/m-3 on 15 Nov 2012, 1306 spores/m-3 on 25 Dec 2012) and 2013/2014 (4575 spores/m-3 on 27 Oct 2013, 4619 spores/m-3 on 3 Nov 2013, 3674 spores/m-3 on 9 Nov 2013, 3521 spores/m-3 on 12 Dec 2013). Results from the qPCR showed that ascospores of L. maculans were released earlier than ascospores of L. biglobosa at Bayfordbury in the 2013/2014 growing season. There were differences in phenotype of isolates amongst ninety-five isolates of L. maculans and forty-eight isolates of L. biglobosa obtained from different sources (phoma leaf spots, upper stem lesions or basal stem cankers) on different cultivars. Cotyledon tests showed that the resistance genes Rlm4, Rlm5, Rlm6 and Rlm7 are still effective in England. Most isolates from phoma leaf spots carried avirulent AvrLm4 (39 isolates; 97.5%), AvrLm5 (39 isolates; 97.5%) and AvrLm6 alleles (36 isolates; 90%) and all 40 isolates carried the avirulent allele AvrLm7 (100%). Fewer isolates from basal stem cankers carried avirulent AvrLm4 (4 isolates, 16.7%) or AvrLm6 alleles (16.7%) but all 24 isolates carried the avirulent AvrLm7 (100%). Fewer isolates from upper stem lesions carried the avirulent AvrLm4 allele (5 isolates; 16.1%), but 15 isolates carried avirulent AvrLm5 (48.4%), 21 isolates carried AvrLm6 (67.7%) and all 31 isolates carried AvrLm7 (100%). By contrast, all isolates were virulent against Rlm1, Rlm2, Rlm3 and Rlm9. This knowledge, together with knowledge about R genes present in current winter oilseed rape cultivars, should be useful to provide recommendations on cultivar selection to growers based on regional frequencies of avirulent alleles of Avr allele genes in the L. maculans populations (races) and improved understanding of the race structure of L. maculans. There is a need to further investigate any R genes that operate against L. biglobosa (possibly from wild brassicas) and to study if any R genes or QR can provide resistance against both L. maculans and L. biglobosa.