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    [23279834 - HortScience] Fusarium Yellows and Turnip Mosaic Virus Resistance in Brassica rapa and B. juncea

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    (23279834 - HortScience) Fusarium Yellows and Turnip Mosaic Virus Resistance in Brassica Rapa and B. Juncea

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    HoarScxence 32(5)'927-930, 1997. Fusarium Yellows and Turnip Mosaic Virus Resistance in Brassica rapa and B. juncea Robert G. Fjellstrom! and Paul H. Williams Department of Plant Pathology, University of Wisconsin, Madison, WI53706 Additional index words. vegetable breeding, disease resistance, oriental Brassicas Abstract. Thirty-seven Brassica rapa L. and B. juncea L. ines from nine subspecies were {ested for their reaction to two pathotypes of Fusarium yellows (Fusarium axysporum ‘Schlecht. f sp. conglutinans (We.) Snyd. & Hans. race 1 and F.o. fp. raphani Kend. & ‘Snyd. A subset of 16 lines from these same vegetable types were tested for their reaction {o four strains of turnip mosaic virus (TuM-C1, C2, C3, and C4). Resistance to both ‘Fusarium pathotypes was widespread in these Brassica subspecies, whereas resistance to any strain of TuMV was uncommon. The broad availability of resistance to Fusarium. yellows and scarcity of resistance to TuMY necessitate different approaches to obtain isease-resistant cultivars. Fusarium yellows, caused by Fusarium ‘oxysporum f. spp. conglutinans and raphant, and turnip mosaic virus (TuMV) are common diseases of Brassica crops (Shatuck, 1992; Walker, 1969). Although the seriousness of Fusarium yellowson 8. rapa syn. B. eampes- tris L.) and B. juncea has not been systemat- cally documented, the importance of this di ease in other erucifers (B. oleracea L. and Raphanus sativus 1; Walker et al, 1958) givesreason for concern about its potential for ‘damaging these Brassica crops, Susceptibility (of B. rapa and B. juncea to TuMV, however, is known to be costly. TMV causes wide- spread devastation within plantings of these vegetables (Faan and Ko, 1957; Nui et al., 1983; Sako, 1981; Wu et al, 1964) Information on resistance to these diseases within the economically important orienta vegetable subspecies of B. rapa and juncea is relatively scarce. Only one subspecies, B. rapa ssp. pekinensis (Lou) Olsson, has been well characterized for the possession of TuMV resistance (Green and Deng, 1983; Provvi- enti, 1980) and only two cultivars have been examined for resistance to yellows (Bosland and Williams, 1988; Ramirez-Villupadua et al, 1985). Thus, aspartofan ongoing program of erucifer pathology research, we undertook anevaluation of Fusarium yellows and TuMV resistance ina set of B. rapa and B. juncea cultivars. This survey of eight B- rapa and one 2B. juncea subspecies for resistance against two pathotypes of F. axysporum and four stains of TUMV provides information con- ‘ceming the prevalence of disease resistance in these erops. [Recaved Tor publication 9 July 1996. Accepted for publication Seb, 1997, The cost of publishing this Paper was defrayed in part by the payment of page hares. ner posal regulations this paper thee fore must be hereby marked advertisement solely (0 indict his fact. ‘Current adress: Pant Molecular Biology Center, Dept of Biological Setences, Neches Univ DeKalb, I. 60115 HorrScience, Vot. 32(5), Avaust 1997 Materials and Methods Pathogen and cultivar sources. Isolates of oxysporum. sp. conglutinans ace | (FOC) and Fo. f. sp. raphani (FOR) were obiained from the collection of PH. Williams, Dept of Plant Pathology, Univ. of Wisconsin, Madi- son (isolates PHWSI and 699, espectively) Isolates of TuMV-C1, C2, C3, and C4 were obtained from R. Provvidenti, Dept. of Plant Pathology, New York State Agr. Expt. Sta, Geneva. Eight subspecies of B. rapa, sspp chinensis L. (pakchoy, tai-tsai, ete.), arachinensis (Bailey) T. & L. (choy sum), ppekinensis (Chinese cabbage), perviridis Bailey (Komatsuna or tendergreen), narinasa Bailey (chizimina, vitamin greens, etc.) ripposinica Bailey (mizuna and mibuna), ‘apifera (Metrg.) Sinsk.(curip), and wis. &L. (hong.-tai-tsai) and one subspecies of B Juncea,ssp.crspfoia Bailey (mustard greens), ‘were obtained from various sources (Table 1). Fusarium disease sereen. Seeds ofthe 37 cultivars to be sereened for resistance to FOC and FOR were sown in silica sand in 30 60 % 8 cm stel pans, grown under continuous itlumination of 400 molars from mixed (1:1 Sylvania coo!-white and Grolux fuores- ‘cent lamps (GTE, Danvers, Mast.) ina 28-5 °C greenhouse, and watered daly. Fusarium isolates were stored in soil at °C. Isolates were grown in SO mLof potato dextrose broth in 250-mL flasks Cultures were incubated at 20°C on a gyrating shaker for 3 t0 5d, then filtered through two ayers ofcheesecloth. The fungal mat collected on top ofthe cheesecloth ‘was rinsed with distilled Water, transferred to a Waring blender containing 50 ml. of dis: tilled water per culture flask and macerated at the highest speed setting for min. Te result ‘ng mixture of hyphae and spores became the primary inoculum forthe disease screen, Fifty 7-d-ld seedlings were inoculated by first removing them from sand using a stel spatula, dipping the roots into water to wash off excess sand, blotting the roots on paper toweling, and submerging the roots in the inoculum suspension for 5 to 10 min. The inoculated seedlings were transplanted into another ste] pan of sandin randomly arranged. roups of 25 (10 groups per pan, two replica- tions per cultivar). The pans were held in Wisconsin soil temperature tanks (Williams, 1981) setat 24+ 1°C. Plants were kept ina 20 £1°C greenhouse, continuously illuminated, as forthe seedling germination, and irrigated daily with 0.5 modified Hoagland solution (Gacabson, 1950). ‘Two weeks after inoculation, the plants were gently removed from the sand and rated for their disease reaction on a scale fom 0 to ‘9(0=nosymptoms; 3 =some stunting of shoot and root growth; 5 = noticeable stunting of shoot and root growah, with leaves turning yellow and roots dark 7 = severe stunting of shoot and root growth, with yellowing and necrosis of leaves, browning and necrosis of roots; 9= death of the plant; Williams, 1981). Plants rated from 0 10 3 on this scale were considered to be resistant and 4 t0 9 10 be susceptible. The percentage of resistant plants and mean disease reaction for each cultivar were then calculated, Twenty-five resistant and 25 susceptible control plants were in- cluded in each screening pan. “Wisconsin Golden Acre" and “Golden Acre’ cabbage for FOC and ‘Fancy Red’ and “White Ieicle"rad- ish for FOR were the resistant and susceptible controls, respectively TuMV disease screen. Seeds of the 16 cultivarstobe screened forresitance toTuMV- C1,C2, C3, and C4 were sown in greenhouse Potting oi in plastic multipot 96 cell trays 39 x 28 x 53 mm/cell), fertilized once weekly With a soluble 20 N-20 P-20 K fertilizer, and ‘grown under natural illumination ina shaded 2845 °C glasshouse ‘The TuMV strains were maintained in tur- nip plants (‘Presto’) isolated in aphid-proof cages. TuMV inocutum was preparedby grind= ing 4 em? of systemically infected turnip leaf witha pestle ina morat containing 5 mL. of (0.03x¢phosphate (K+) bufferatpH 7.0and0.2 'g0f 200 mesh (72 um particle size carborun- dum Fony-cight 14-d-old plants of each euli- var were randomized into four replications of 12 plantsteplication/ultivar, lightly dusted With 200 mesh carborundum, and inoculated by rubbing thefirsttwo true leaves with inocu- lum. Inoculated plants were covered with wetted newspaper for 20 h after inoculation Plants were observed for mosaic symptoms 10, aftr the inital inoculation and, at this time, symptomless plants werereinoculatedontheir third and fourth re leaves. Ten and 20 d after the second inoculation, plants were rated for their disease reaction ona scale from 010 9 (0 = no symptoms; 1 = symptoms restricted 10 ‘small chlorotic and necrotic lesions on the inoculated leaves; 3 = chlorotic or necrotic lesions on the inoculated leaves plus slight :motling on new growth; 5 = 3 + increased ‘mottling and chlorosis on new grovth, no leaf deformation; 7=3 + severe chlorotic moting, 60'%) aver- age esistant frequencies tothe two pathotypes. ‘A >S0% average frequency of resistance against one pathotype, but not against the other, occurred inthe B.rapassp.parachinen- sis, sp. perviridis, ssp. nipposinica, and B. Juncea ssp. erispiolia cultivars. The B. rapa ‘sp. nipposinica cultivars wer relatively Su8- ceptible (22.0% average resistance frequency) to POC and the B. rapa ssp. parachinensis, pervridis, and B. juncea ssp. crispifoliaculi- ‘vars wererelatively suscepble (36.0%, 26 0%, and 22.3% average resistance frequencies, respectively) 0 FOR, ‘Despite the relatively common occurrence ‘of resistance, the cultivars were quite hetero- ‘eneous in their disease reaction to the Fusarium pathotypes. Only five of the 35 cultivars were uniformly (295%) resistant to FOC and only wo were uniformly resistant to FOR, None were uniformly resistant to both pathotypes TuMfV tests. Resistance against any ofthe strains of TuMV (Cl, C2, C3, and C4) was uncommon (Table 3). Out of 64 total cultivar- strain combinations tested, there were only four combinations that showed a cultivar hav- ing a SO% or higher frequency of resistant plants to any one strain ofthe virus. Cultivars having >59% frequency of resistant plans oc- curred in only 11 of the 64 combinations. “Souther Giant Cured’, of B. juncea ssp. crspiflia wasthe only cultivar having >SO% frequencies of resistant plants against more than one strain of TuMV (having 72% and 181% frequencies of resistance aginst strains C1 and C2, respectively), Resistance against any one strain of the virus generally had low correlation (absolute value F< 0.16) with resistance to any other 928, strain, The highest coreltions were between CH and C2 resistance and Cl and C3 ress tance, being 0.566 and 0.542, respectively ‘Thecorrelationfresistance frequency tomean disease reaction was only moderately high ( 0153). Resistance frequencies >3% to the various TuMV strains were found among cul- tivats of B. rapa sp. pekinenss, ssp. chinen- sis, and ssp. rapifea. Cultivars belonging to the other B. rapa subspecies tested had

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