Angular Leaf Spot of Cucumber in Japan

YASUMASA WATANABE* and AKIRA OHUCHI**

* National Institute of Agricultural Sciences (Yatabe,Ibaraki, 305 Japan) * Hokuriku National Agricultural Experiment Station (Inada, Joetsu,Niigata, 943-01 Japan)

Cucumber angular leaf spot is one of the culture, Forestry and Fisheries organized a most prevalent diseases of cucumber (cucur­ co-operative research program "Studies on bit) in Japan. The affected fields covered the integrated control of bacterial diseases in 6,240 hectares in 1980, accounting for about cucurbits"23> which lasted four years from a half of the total cucumber-planted area in­ 1976 to 1979, in order to establish a combined cluding plastic house cultivation. cultural and chemical control procedure. The The first incidence of cucumber angular authors will describe outline of the results leaf spot in Japan was reported by Tominaga obtained in this research program. & TsuchiyaH> in 1957. Since Mukoo et al.OJ recognized its severe incidence in Kanto dis­ Causal bacterium trict in 1971, it has spread throughout Japan. The causal bacterium not only produces Ohuchi et al.O> examined more than one spotted lesions on cucumber leaves ( Plate 1) hundred isolates collected from affected cu­ but also affects cucumber fruit (Plate 2) , fol­ cumber plants which were distributed in the lowed by entire decay of the fruit. various parts of Japan and identified 110 The disease was so severe in cucumber­ isolates as Pseuclomonas syringae pv. Zachry­ growing districts that the Ministry of Agri- mans (Smith & Bryan 1915) Young, Dye &

Plate 1. Typical symptom of cucumber angular leaf spot on leaf 113

isolates by needle pricking on cucumber fruit segments and incubating in moist chamber at 24 °C. P. syringae pv. lachrymans was detected by conspicuous ooze exudation on the fruit segments (Plate 3). Although several other bacteria were capable of exudating bac­ terial ooze, they were not pathogenic to cucum­ ber, except P. syringae pv. mellea and P. syringae pv. tabaci. These two bacteria were distinguished from P. syringae pv. lachrymans by yellow haloes surrounding lesions on cu­ cumber leaves.

Disease cycle

The disease has spread throughout Japan in these several years. Such rapid spread of the disease suggests that the causal bacterium should be carried by infested seeds, as was proved by Carsner in 1918.2> Exact seed trans­ mission of the disease in Japan was con­ firmed through the experiment using the seeds obtained from naturally infected fruit of cucumber in the open field for seed pro­ duction.7,Hil The rate of seed transmission was as low as less than one per cent. Less than one per cent of seed transmission, however, was proved to be enough for overall incidence of angular leaf spot in a nursery bed, when Plate 2. A fruit affected by cucumber angular leaf spot disease, the bed was kept under high moisture con­ showing a clear secretion of dition. Hi) tea1·-like droplets Transmission of the disease through in­ fested soi l has been suspected in the fields Wilkie 1978 on the basis of their bacteriologi­ where angular leaf spot broke out every year. cal properties and pathogenicity to host plants. Soil transmission, however, has not been con­ On the contrary, the isolates from the lesions firmed clearly yet. Ogawa et al.Ill showed that characterized by blighting of leaf margin the disease symptoms appeared on the coty­ were identified as Pseudomonas viridi/lava ledons of cucumber seedlings when the causal (Burkholder 1930) Dowson 1939 (Table 1) .10 l bacteria were introduced in the soil above This bacterium was responsible for the soft sowing level, suggesting direct contact of rot of cucumber fruit which was often ob­ cotyledons with causal bacteria during ger­ served in the cucumber fields affected by mination of seeds. The longivity of causal angular leaf spot, and was advocated as an bacteria in soil was examined at several additional causal agent of marginal blight of laboratories in Japan using indicator plant cucumber. 12> method.23> The longivity was dependent on Ohuchi et aJ.1 1> developed successfully a seasons when diseased cucumber vines were rapid method for distinguishing P. syringae buried in soil. In summer the causal bac­ pv. lachrymans from other bacteria. The terium died within 10 to 20 days, whereas method consists of inoculating with bacterial longivity of causal bacterium was 3 to 4 114 JARQ Vol. 17, No. 2, 1983

Table 1. Major bacteriological characteristics of two species of pathogenic bacteria

Pseudomonas syringae pv. lachrymans Pseudomonas viridiflava Property (110 isolates from affected cucumber (45 isolates from affected cucumber plants characterized by angular plants characterized by blighting spotted lesion on leaf) of leaf margin) Gram stain 0-F test 0 0 Flagella (polar) 1-5 >5 Poly-,8-hydroxybutyrate accumulation Fluorescent pigment + + Growth at 37°C Growth in Cohn's solution + Arginin dehydrolase activity Oxidase activity ,8-glucosidase activity + + Tyrosinase activity Hydrolysis of casein + Hydrolysis of cotton oil Production of levan + Nitrate reduction Liquefaction of gelatin + + Utilization of tartrate + Utilization of sucrose + Softrot activity to potato + Hypersensitivity to tobacco + +

P late 3. Conspicuous ooze exudation on the fruit segments inoculated with P. sy1·-inycw pv. laclwymans by needle pricking 115

Plate 4. Effect of atmospheric humidity on the development of lesions of cucumbet' ang·ular leaf spot A: characteristic angulal' spotted lesions developed under high moistme condition ( t'elative humidity of 90-949',, or above) B: minute lesions developed at 85% relative humidity

months in winter.4l These results indicate Predisposing factors that soil transmission of the disease is likely to occur in winter, if the period of fallowing Angular leaf spot of cucumber in Japan is in cucumber fields is within 3 to 4 months.Gl sevel'e in plastic house cultivation in winter as well as in open field cultivation in the northern part of Japan. Thus it. seems that 116 JARQ Vol. 17, No. 2, 1983 a condition which combines low temperature leaf spot of cucumber by means of controlling with high humidity is favorable for the severe atmospheric humidity at night in plastic occurrence of the disease. Umekawa & Wata­ houses. The dehumidifiers equipped in plastic nabe10> investigated the effect of temperature houses proved to be effective in controlling and atmospheric humidity on the occurrence the incidence of angular leaf spot as well as of angular leaf spot of cucumber. The lesions for keeping atmospheric humidity in plastic developed severely at 15° to 25°C under a houses below 90%.19) Only slight marginal moisture saturated condition. Relative hu­ blight lesions were developed on leaves of midity of 90 to 94% or above was essential cucumber grown in the equipped houses for the development of characteristic angular (Plate 5) . lesions, whereas only minute lesions developed Rainfall affects the incidence of cucumber at 85 % relative humidity (Plate 4) . The angular leaf spot in open-field cultivation in duration of misting after spray of bacterial northern part of Japan. Umekawa et a1.1 s, suspension had a marked effect on lesion revealed that the rain water dripping from development. Twenty-four hours duration of the diseased leaves contained the causal misting after inoculation was necessary for bacteria at the concentration of 1or.-100 cell the development of characteristic lesions, per ml of rain drop, and sheltering of cucum­ whereas minute lesions were produced when ber vines by roof of plastic film was able to inoculated plants were kept under dry con­ reduce the development of angular leaf spot dition (relative humidity of 40 to 60 %) . lesions. They concluded that cultivation of cu­ These results coincide substantially with those cumber avoiding l'ain under plastic roof was of Williams et a1.21 J effective on the control of angular leaf spot. Moisture content in soil also affects the development of lesions on leaves. Experiments Table 2. Effect of duration of misting at night conducted in a green house provided the re­ on the development of lesions of cucum­ sults that the cucumber plants grown in the ber angular leaf spot under a diurnal soil with higher moisture content were af­ alternation of humid night and dry day. fected more severely by angular leaf spot.l=ll tim e Diurnal alternation Varietal resistance Duration Duration Disease index Type of lesion of misting of dry condi- Most commercial varieties of cucumber now (hr) tion (hr) in use in Japan are susceptible to angular 0 24 1. l Minute lesion leaf spot, whereas 'Kurume-ochiai H and 1 23 l.l Minute lesion Maji-midori' were proved to be tolerant 3 21 2. l Minute lesion 6 18 3. l Developed lesion through the co-operative research prngram.2:q 12 12 3. 0 Developed lesion Umekawa & Watanabe17l established the meth­ od of assaying the resistance to angular leaf spot during a seedling stage and evaluated that 'Saitama-ochiai No. 4, Shogoin-aonaga­ Atmospheric humidity in fields or plastic fushinari, Kariba-fushinari and Saclo-1·isshu' houses, however, varies rithmically from were as resistant as 'Poinsett' which was humid condition at night to dry condition in rated as highly resistant variety in USA.1J the daytime. Under such an alternating hu­ In order to develop the resistant varieties to midity, the duration of less than 3 hr of moist angular leaf spot, Kawaicle3> selected 'Sai­ condition at night could not produce angular tama-ochiai No. 4, Sakata and Suyo for par­ lesions but minute lesions on cucumber leaves ents and established the populations carrying (Table 2). These results suggest that it may l'esistant genes. be possible to reduce the incidence of angular 117

Plate 5. Effect of dehumidifiers equipped in a plastic house on the incidence of cucumber angular leaf spot in the plastic house. A. severe incidence of cucumber angular leaf spot in a standard plastic house B: very few lesions on the leaves of cucumber grown in a dehumidifier-equipped plastic house C: slight marginal lesions on the leaf of cucumber grown in the dehumidifier-equipped plastic house 118 JARQ Vol. 17, No. 2, 1983

od. Ann. Phytopcithl. Soc. Jvn., 46, 83- 84 Seed disinfection and spray of (1980) [Abstract. In Japanese]. chemicals 5) Kuniyasu, K. & Takeuchi, S.: Cucumbe1· angular leaf spot, possibility of passing winter and summer through soil. Proc. Various methods of seed disinfection against Ifonsai Plant Prot. Soc., 22, 80 (1980) [Ab­ angular leaf spot were examined through the stract. In Japanese]. co-operative research program.2Hl Among the 6) Mukoo, H. et al. : Angular leaf spot of methods examined, soak treatment of infested cucumber. Ann Phytopcithl. Soc. )Jm., 37, seeds in 1 :20 dilution of sodium hypochlorite 368 (1971) [Abstract. In Japanese]. 7) Nagai, Y. & Fukatsu, R.: Occurrence and solution (active Cl : 4 %) for 20 min was seed transmission of cucumber angular leaf proved to be effective against seed transmis­ spot. Proc. Kanto-Tosan Plant Prot. Soc., sion of angular leaf spot from a lot of experi­ 17, 47 ( 1970) [Abstract. In Japanese]. mental results. Soak t1·eatment in 0.25 to 8) Ogawa, IC et al.: Soil transmission of cu­ 0.5 % solution of "Chemiclone G" (Calcium cumber angular leaf spot. Ann. Phytovathl. Soc. Jpn., 44, 59 ( 1978) [ Abstract. In J apa­ hypochlorite, active Cl: 70%) for 60 min was nese]. also effective. They were recommended for 9) Ohuchi, A. et al.: Bacteriological prop­ practical use because of their little phyto­ erties and pathogenicity of Pseudomonas toxici ty to seed germination. Soak treatment lachrymans, the causal agent of angular in streptomycine solution was not practical leaf spot of cucumber, from Japan. Bull. Nat. Inst. Agr. Sci., Ser. C 34, 1- 13 (1980) because of wide distribution of streptomycil1- [In Japanese with English summary]. resistant strain of Pseudo11ionas syringae pv. 10) Ohuchi, A. & Ezuka, A.: Pseudomon

Phytovathl. Soc. Jpn., 45, 560-561 (1979) 20) Watanabe, Y. : Chemical control of cucum­ [Abstract. In Japanese]. ber angular leaf spot. Kongetsii-no-Noyaku, 17) Umekawa, M. & Watanabe, Y.: Varietal 19 (11), 86- 90 (1975) [In Japanese]. resistance to angular leaf spot of cucumber 21) Williams, P. H. & Keen, N. T. : Relation of and assaying method of resistance during cell permeability alterration to water con­ seedlings. Bull. Veg. & Ornam. C1·ops Res. gestion in cucumber angula1· leaf spot. Sta. hm., Ser. B 3, 67-73 (1980) [In Japa­ Phytopathl., 57, 1378-1385 (1967). nese with English summary]. 22) Yano, H. et al.: Drug-resistance of cucum­ 18) Umekawa, Y. et al.: Facilitative effect of ber angular leaf spot bacterium, Pseudo1no­ rainfall on the transmission of the pathogen nas lachrymans (Smith et Bryan) Carser. and the development of angular leaf spot Ann. Phytopathl. Soc. Jpn., 44, 334-336 of cucumber. Ann. Phytopathl. Soc. Jpn., (1978) [In Japanese with English sum­ 47, 346-351 (1981) [In Japanese with Eng­ mary]. lish summary]. 23) Anonymous: Integrated control of bac­ 19) Umekawa, M. & Watanabe, Y. : Relation terial disease in cucurbits. Interim report. of temperature and humidity to the occur- Agriculture, Forestry and Fisheries Re­ 1·ence of angular leaf spot of cucumber search Council, Japan (1979) [In Japa­ grown under plastic house. Ann Phytopathl. nese]. Soc. Jpn., 48, 301-307 (1982) [In Japanese with English summary]. (Received for publication, September 13, 1982)