Norihito Yamauchi

Physiological races and vegetative compatibility groups of butterhead lettuce isolates of Fusarium oxysporum f. sp. lactucae in Japan

Races were identified among butterhead lettuce isolates of Fusarium oxysporum f. sp. lactucae collected from three geographical areas of Hokkaido, Shizuoka, and Fukuoka in Japan by inoculation tests using Fujinaga’s race differential cultivars of lettuce (i.e., Patriot, Costa Rica No. 4, and Banchu Red Fire). Eighteen isolates from Shizuoka and Fukuoka were designated race 3, with two unknown vegetative compatibility groups (VCGs) that differed from Ogiso’s VCG 1 and 2. These two new VCGs were obtained from both Shizuoka and Fukuoka. On the other hand, three isolates from Hokkaido were classified as race 1 and identified as VCG 1, which represents a VCG of crisphead isolates from Nagano.

Pathogenicity Groups in Fusarium oxysporum f. sp. lactucae on Horticultural Types of Lettuce Cultivars

Isolates of Fusarium oxysporum f. sp. lactucae obtained from six localities in Japan were divided into three patho-genicity groups. Group 1 was highly pathogenic to lettuce cultivars of crisphead and red leaf types and was less pathogenic to butterhead and green leaf type cultivars. Group 2 was highly pathogenic to butterhead type and less pathogenic to crisphead and leaf types. Group 3 was less pathogenic to all lettuce types than groups 1 and 2. These results indicated pathogenic differentiation in F. oxysporum f. sp. lactucae, roughly relating to horticultural types of host lettuce cultivars.

Development of sequence tagged site markers to identify races of Fusarium oxysporum f. sp. lactucae

By random amplified polymorphic DNA (RAPD) analysis of the representative isolates of each race of Fusarium oxysporum f. sp. lactucae, RAPD fragments of 0.6, 1.6, and 2.9 kb were obtained. The 0.6-kb RAPD fragment was common to the representative isolates of all three races. Amplification of the 1.6- and 2.9-kb fragments were unique to the isolates of races 1 and 2, respectively. Sequence tagged site (STS) marker FLA0001, FLA0101, and FLA0201 were generated from the 0.6-, 1.6-, and 2.9-kb RAPD fragments, respectively. Polymerase chain reaction (PCR) analysis showed that FLA0001 was common to all 49 isolates of F. oxysporum f. sp. lactucae. FLA0101 was specifically generated from all 23 isolates of race 1 but not from races 2 or 3. FLA0201 was specifically amplified from all 12 isolates of race 2 but not from races 1 or 3. In two isolates of F. oxysporum f. sp. lactucum, PCR amplified FLA0001 and FLA0101 but not FLA0201. On the other hand, these STS markers were not detected from isolates of five other formae speciales. Because these STS markers were not generated from isolates of other plant pathogenic fungi, bacteria, or plant materials examined in this study, PCR analysis combined with the three STS markers should be a useful means for rapid identification of races of F. oxysporum f. sp. lactucae.

A New Race of Spinach Downy Mildew in Japan

A new race of spinach downy mildew caused by Peronospora effusa occurred in Fukui, Japan. The fungus was capable of affecting spinach cultivars resistant to races 1, 2, 3 and 4, but not some other cultivars. Thus, the fungus had different pathogenicity from race 3 and race 4 of the pathogen and was considered to be a new race of spinach downy mildew in Japan.

Recent races of spinach downy mildew pathogen Peronospora farinosa f. sp. spinaciae in Japan

Isolates of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, collected at seven sites in Japan from December 2013 to March 2016, were identified as belonging to races 8, 10, 12 and 13, and one was unauthorized. Races 10, 12, 13 and the unauthorized were first found in Japan.

First report of spinach downy mildew caused by race Pfs:8 of Peronospora farinosa f. sp. spinaciae in Japan.

The race of field isolates of Peronospora farinosa f. sp. spinaciae (Pfs), causal agent of spinach downy mildew, were identified using race-differential cultivars. One isolate was similar to race Pfs:6. Three isolates were identified as race Pfs:8, the first time the race has been reported in Japan as far as we know. The differential reaction caused by the other two isolates did not match any known to be caused by races Pfs:1 through Pfs:11; thus, this strain appears to a new pathogenic strain in Japan.

Characterization of biotin-autotrophic isolates derived from naturally occurring auxotrophic race 2 isolates of Fusarium oxysporum f. sp. lactucae

Race 2 isolates of Fusarium oxysporum f. sp. lactucae have been recognized as biotin auxotrophs and consequently have restricted growth on Puhallas minimal medium (MM), which contains no biotin. Biotin-autotrophic isolates were raised from race 2 isolates through cultural mutation that grew as well on MM as they did on MM supplemented with biotin. These autotrophs were identical to the parental isolates in pathogenicity on race differential cultivars of lettuce (Patriot, Banchu Red Fire, and Costa Rica No. 4), and thus were designated as race 2. A vegetative compatibility test indicated that the autotrophic isolates fell into the same vegetative compatibility group as the parents. Culture filtrates of the autotrophs allowed abundant growth of the parental auxotroph on MM, and, through a competitive enzyme-binding assay, biotin was detected in the culture filtrates. These results suggest that biotin auxotrophy in the natural race 2 isolates has no direct relation to pathogenicity, qualitatively defined as physiological race, or to vegetative compatibility.

New races of the spinach downy mildew pathogen, Peronospora farinosa f. sp. spinaciae, in Japan in 2017

Six isolates of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, collected from Saitama and Ibaraki Prefectures in 2017 were identified. Three isolates collected from Saitama Prefecture in January differed from all the known races described by the International Seed Federation but their virulence was similar to that of strain UA1014 sampled in the USA in 2014. Isolate Iba17-2, collected from Ibaraki Prefecture in April, was identified as race 10; isolates Iba17-1 and Iba17-3, collected from the same region in February and May, did not match any known races and were coded as novel races, J17-1 and J17-2, respectively.