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1 Res. Plant Dis. 16(2) : (2010) Research in Plant Disease The Korean Society of Plant Pathology z w t w sƒ Á½ Á Á yá * w yw yw l Development of Effective Screening Method and Evaluation of Radish s for Resistance to Fusarium oxysporum f. sp. raphani Song-Yi Baik, Jin-Cheol Kim, Kyoung Soo Jang, Yong Ho Choi and Gyung Ja Choi* Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology, Daejon , Korea (Received on June 25, 2010; Accepted on July 6, 2010) To establish the efficient screening method for resistance of radish to F. oxysporum f. sp. raphani, we investigated the development of Fusarium wilt of two radish cultivars, Songbaek (susceptible) and Tokwang (moderately resistant), according to several conditions such as inoculation methods, inoculum concentrations, and dipping periods of radish roots in spore suspension. By infected soil and soil-drenching inoculation methods, Fusarium wilt did not occur on the seedlings of both cultivars. In root dipping inoculation method using cut or non-cut roots of radish plants, the cut roots were easily infected by the pathogen than non-cut roots. And the disease development of two cultivars represented significant difference in non-cut root method. On the other hand, disease severity of Fusarium wilt on radish seedlings according to inoculum concentration increased in a dose-dependant manner, regardless of dipping periods. Using screening method established from the results, the 41 commercial radish cultivars were evaluated the degree of resistance to F. oxysporum f. sp. raphani. Among them, 6 radish cultivars were resistant, 22 cultivars were moderately resistant, and 13 cultivars were susceptible to Fusarium wilt. Keywords : Breeding, Fusarium wilt, Radish yellows, Raphanus sativus, Resistant (Raphanus sativus) l ƒ xk, w, w (t, 1989). ù 5 wù ,192 ha 533,210m ( t, 2007). û t x y ƒ ƒ w ù,» w ƒ w wƒ ƒwš. ¾ w w k 19 š (w wz, 2009). j F. oxysporum f. sp. raphani ³ m ü z s x w» *Corresponding author Phone) , Fax) ) [email protected] { k w ƒ w. š y w yw z s w» e w k (van Peer, 1988) e s San Benito White Chinese Winter Radish s š, z 1946 g Waukesha s š z x ƒ wš (Pound, 1959; Pound Fowler, 1953). ù l w wš ù, 1987 š (û, 1994). ù 6 ~7» w l wì y w š y» ƒ w, w w t š w ƒ y ù y, z

2 w t ³ w (, 1990;, 2003). w» w w t w ù, z q, z,, m š (, 2002). w m z m ³ û, ü ³ ƒ (w yxz, 2008) y w ƒ ey ƒ ƒw z w ³ š. ú w t ƒ y ey š. x w t, w l, mg, k, g š z q wš ù, w ³ w š f w š. w z w v. z w y wš,, e w w, y w w qt 41 w y w.. w y w» w x mÿ ( w l) w sp(sp m 20 ml, ( ) ) m 5y( ) š 1 q w (25±5 C) 14 w o x w. q t w x z w t q wš 8 t w œ w 33 t w w w x w. 5 8 sp(sp m 70 ml, ( ) ) m 5y š 1 q w 7, 10, 14 w z x w.. j F. oxysporum f. sp. raphani KR1 ³ l, ³ potato dextrose agar(becton, Dickinson and z w t w sƒ 149 Co.) e w 25 o C w» 7 w ³ l ³ ƒ malt extract broth(becton, Dickinson and Co.) wš 25 o C k rpm k w. w KR1 ³ 4 ³ wš, Ÿwx w hemocytometer w s ( x s ) d w. ³ conidia/mlƒ 7 w w. x ƒƒ , , , conidia/ml 7 w, , conidia/pot, m m ml ƒ 4 w. ³. m ³ w w x w w w z s xk 2 e w w. w 5 8 sp m 5y w w. e w ƒ w 2 cm û»š z s xk e w. w e s xk ƒƒ 1, 2, 4, 8, 16, 24 e w w. 5 8 sp w w s xk sp 5 ml w w. m ³w m 5y 100 ml s xk 5 ml š w m 5 8 sp(sp m 70 ml) š sp 1 q w w.. w 1 25 o C w z (25±5 C) 3~4 o w w. w z w. 0=, 1= w ù š, 2= w š, 3= w š yyw, 4= w š w y w š ù, 5=š 5 w, s³ ƒ 1.0 w w, 1.0~2.5 w, 2.5 q w. qt w x 10 2z w, w x 5 2z w. SAS(SAS Institute, Inc., 1989, Cary, NC) v w ANOVA w, s³

3 150 Á½ Á Á yá w Duncan's multiple range test(p=0.05) w. š. mÿ j» w,» Fusarium w w m, š s xk e w xw. m mÿ w. (1990) m:vermiculite=1:1 yw m m ml ƒ 4 F. oxysporum f. sp. raphani s xk w m q w x ww š šw ù, x m ml w m q w w, w. m w ww ƒ. š ³ w w sp s w 8 xw» w (Table 1). (1996) (F. oxysporum f. sp. conglutinans) w ³ w, m e š šw. ù w x w, w w ww ƒ. Table 1. Occurrence of Fusarium wilt in two radish cultivars after soil drenching of spore suspension of Fusarium oxysporum f. sp. raphani KR1 a wr, e üš w, mÿ (Table 2). ù s xk e w xw ù, mÿ û ùkþ. w w w t y w ùkü ww. Table 2. Development of Fusarium wilt in two radish cultivars when cut and non-cut roots were dipped in the spore suspension of Fusarium oxysporum f. sp. raphani KR1 y Method Tokwang Songbaek Cut root 4.9±0.1a z 4.6±0.3a Non-cut root 2.6±0.3b 4.5±0.4b y 14-day-old seedlings of two radish cultivars were inoculated with F. oxysporum f. sp. raphani KR1 by dipping the roots of radish seedlings in spore suspension of conidia/ml for 2 hr. The z Each value represents the mean disease index±standard deviation of two runs with five replicates each. Values in the labeled with the same letter are not significantly different in Duncan s multiple rage test at P = Inoculum (conidia/pot) Plant growth stage (after sowing) 7 days 10 days 12 days Songbaek 0 b Tokwang Songbaek Tokwang a Each seedling of two radish cultivars was inoculated with F. oxysporum f. sp. raphani KR1 by drenching the spore suspensions on soil of each pot. The infected plants were incubated in humidity chamber at 25 o C for 24 hr and then transferred to a greenhouse at 25±5 o C. After 3 weeks, disease severity of the radish seedlings was investigated. b Each value represents the mean disease index of two runs with five replicates each. Fig. 1. Fusarium wilt occurrence of two radish cultivars according to inoculum concentration. 14-day-old seedlings of two radish cultivars were inoculated with Fusarium oxysporum f. sp. raphani KR1 by dipping the roots of the radish seedlings in each spore suspension for 2 hr. The infected plants were incubated in humidity chamber at 25 o C for 24 hr and then transferred to a greenhouse at 25±5 o C. After 3 weeks, disease severity of radish seedlings was investigated. Each value represents the mean of two runs with five replicates each. Values in the labeled with the same letter are not significantly different in Duncan s multiple rage test at P = 0.05.

4 . mÿ t ƒ ƒw ƒw (Fig. 1) conidia/ml s xk 7 e w mÿ ƒƒ ùkü, conidia/ml 4.2, mÿ 3.4. Kuc(1987, 1994) w š ³ w ƒ» ù w w yw w š šw. w j w conidia/ml s xk e w w ƒ. e. mÿ ³ s xk ƒƒ 1, 2, 4, 8, 16, 24 e w w,. w e ¼ w ƒw (Table 3). 1 e w e w w mw v ƒ. (2003) y w m x s xk 2 e w xw. š q s ü w» w, ³ w s xk e 2 w q (Agrios, 2005). F 1 t w. y w w w x q 41 t w w. z w t q wš 8 t Table 3. Fusarium wilt development of two radish cultivars according to root dipping period y z w t w sƒ 151 Incubation period (hr) Songbaek 4.8a z 3.6ab 3.6ab 2.2b 3.0ab 3.0ab Tokwang 4.6a 3.4ab 4.2ab 4.0ab 3.2ab 3.4ab Table 4. Resistance degree of the 41 commercial radish cultivars to Fusarium wilt caused by Fusarium oxysporum f. sp. raphani KR1 a Company Trait Disease index A01 A YR b 0.4±0.14 c B01 B YR 0.6±0.42 C01 C 0.7±0.00 D01 D 0.9±0.14 E01 E YR 0.9±0.28 C02 C 1.0±0.21 C03 C 1.1±0.35 C04 C 1.1±0.35 D02 D 1.1±0.00 C05 C 1.1±0.00 E02 E 1.2±0.21 B02 B 1.3±0.35 B03 B 1.3±0.21 B04 B YR 1.3±0.07 C06 C 1.3±0.28 F01 F YR 1.4±0.35 B05 B YR 1.4±0.49 B06 B YR 1.4±0.28 B07 B 1.4±0.42 B08 B 1.6±0.28 D03 D 1.7±0.14 B09 B YR 1.8±0.21 A02 A 1.9±1.06 B10 B 2.0±0.14 A03 A 2.1±0.71 A04 A 2.2±0.07 A05 A 2.2±1.13 C07 C 2.5±0.28 A06 A 2.8±1.56 B11 B 2.8±0.42 D04 D 2.9±0.35 C08 C 2.9±0.21 C09 C 3.2±0.49 D05 D 3.2±0.42 C10 C 3.2±0.28 B12 B 3.3±0.49 C11 C 3.4±0.14 A07 A 3.5±0.07 D06 D 3.6±0.71 B13 B 3.9±0.92 C12 C 4.0±0.00 y 14-day-old seedlings of two radish cultivars were inoculated with Fusarium oxysporum f. sp. raphani KR1 by dipping the roots of radish seedlings in spore suspension of conidia/ml. The z Each value represents the mean disease index of two runs with five replicates each. Values in the labeled with the same letter are not significantly different in Duncan s multiple rage test at P = a 14-day-old seedlings of each radish cultivar were inoculated with F. oxysporum f. sp. raphani KR1 by dipping the roots of radish seedlings in spore suspension of conidia/ml for 2 hr. The b Resistant cultivar to Fusarium wilt supported by each seed company. c Each value represents the mean±standard deviation of two runs with ten replicates each.

5 152 Á½ Á Á yá 3 t (38%) w ùkþ ù, 5 t (62%) w ùkü (Table 4). wr, w œ w 33 t 3 t (9%) w w ùkü, 17 t (52%) w ùkþ. 13 t (39%) w. q t 41 t w 6t (15%) w, 22t (54%) w, š 13t (31%) q t 70% ƒ w w š. w w y w» w, e, ( ) mÿ ( w ) w. m, e xw, m w w. e, w, t ùkþ. s xk e w (1 ~24 ), ƒw ƒw. ù s xk s ƒ ƒw ƒw. w q 41 t w w. 14 w conidia/ml s xk 7 2 e w xw, 6 t (15%) w, 22 t (53%) w š 13 t (32%). t» ( y : y) w. š x Agrios, G. N Parasitism and disease development. In: Plant Pathology. pp Academic Press, New York. Ÿ,, ½,,, w», ³ 2 j y : y (Radish Yellows), ³ w j y. w yw. pp w yxz e p. w wz w 5q. pp Kuc, J Plant immunization and its applicability for disease control. In: Innovative approaches to plant disease control, ed. by I. Chet, pp John Wiley, New York. Kuc, J Induced systemic resistance, a non-pesticide technology for disease control in plant. In: Proc. 4th. Nat. Conf. Pesticides, ed. D. L. Weigmann, pp Blacksburg, Virginia., xûy, ³, w, ½ ³ y w m, w št, y t. ( ). 130 p. û x Fusarium w w. û w w », t pp t , m. pp Pound, G. S Red Prince is new radish. Wis. Univ. Agric. Exp. Sta. Bull. 538: 93. Pound, G. S. and Fowler, D. L Fusarium wilt of radish in Wisconsin. Phytopathology 43: tx, wƒ. w. 610 p. y, ½, x y w t. w wz 28: van Peer, R., Xu, T., Rattink, H. and Schippers, B Biological control of carnation wilt caused by Fusarium oxysporum f. sp. dianthi in hydroponic system. ISOSC Proc