scholarly journals Molecular identification and characterization of binucleate Rhizoctonia spp. associated with black root rot of strawberry in Serbia

2018 ◽  
Vol 33 (2) ◽  
pp. 97-107 ◽  
Author(s):  
Mira Vojvodic ◽  
Brankica Tanovic ◽  
Milica Mihajlovic ◽  
Petar Mitrovic ◽  
Ivana Vico ◽  
...  

Strawberry production is a popular, fast-growing agricultural business in Serbia. Its cultivar selection has been changing fast, following market demands. One of the limiting factors of strawberry production worldwide is black root rot, primarily caused by binucleate Rhizoctonia. Recently, outbreaks of black root rot of strawberry have occurred in Serbia and the estimated disease incidence was up to 30%. Isolates of binucleate Rhizoctonia AG-A were recovered from symptomatic strawberry plants, and characterized on the bases of morphological, molecular and pathogenic features. Despite their uniform morphological characteristics, the isolates demonstrated genetic variability within ITS rDNA, grouping into three different phylogenetic sub-clusters which comprise AG-A isolates originating from Italy, Israel, Japan and the USA. The binucleate Rhizoctonia AG-A from Serbia exhibited uniform virulence on strawberry after inoculation of daughter plants and detached leaf petioles, as well as on seedlings of bean, carrot and sunflower, while they were non-pathogenic to wheat, maize, tomato, pepper, tobacco, cucumber, lettuce, peas, cabbage, rapeseed and sugar beet.

Plant Disease ◽  
2008 ◽  
Vol 92 (9) ◽  
pp. 1368-1368 ◽  
Author(s):  
S. T. Koike

In 2005 and 2006, field-grown iceberg lettuce (Lactuca sativa) in California's coastal Salinas Valley (Monterey County) was affected by a previously unreported disease. Symptoms were observed on iceberg lettuce at the post-thin rosette stage (8 to 12 leaves). Plants were stunted and slightly chlorotic. Fine feeder roots had numerous, small (4 to 8 mm long), elongated, dark brown-to-black lesions. Larger secondary roots and taproots lacked lesions. No vascular discoloration was present. Isolations from root lesions consistently resulted in gray fungal colonies that formed catenulate, cylindrical, thin-walled, hyaline endoconidia and catenulate, subrectangular, thick-walled, dark aleuriospores. The fungus was identified as Thielaviopsis basicola (2). Conidial suspensions (5.0 × 105) of eight isolates from iceberg lettuce were used for pathogenicity tests. Iceberg cv. Ponderosa and romaine cv. Winchester were grown for 3 weeks in soilless peat moss rooting mix. Roots of 20 plants per cultivar were washed free of the rooting mix and soaked in conidial suspensions for 5 min. Plants were repotted and grown in a greenhouse. Control plant roots were soaked in sterile distilled water (SDW). After 3 weeks, inoculated iceberg exhibited slight chlorosis in comparison with control plants. Feeder roots of all iceberg plants inoculated with the eight isolates exhibited numerous black lesions and T. basicola was reisolated from these roots. Romaine lettuce, however, did not show any foliar symptoms. Small segments of roots had tan-to-light brown discoloration and T. basicola was occasionally reisolated (approximately 40% recovery). Roots of control iceberg and romaine showed no symptoms. Results were similar when this experiment was repeated. To explore the host range of T. basicola recovered from lettuce, two isolates were prepared and inoculated as described above onto 12 plants each of the following: iceberg lettuce (cv. Ponderosa), bean (cv. Blue Lake), broccoli (cv. Patriot), carrot (cv. Long Imperator #58), celery (cv. Conquistador), cotton (cv. Phy-72 Acala), cucumber (cv. Marketmore 76), green bunching onion (cv. Evergreen Bunching), parsley (cv. Moss Curled), pepper (cv. California Wonder 300 TMR), radish (cv. Champion), spinach (cvs. Bolero and Bossanova), and tomato (cv. Beefsteak). Control plant roots of all cultivars were soaked in SDW. After 4 weeks, only lettuce and bean roots had extensive brown-to-black lesions, from which the pathogen was consistently resiolated. Roots of cotton, pepper, spinach, and tomato had sections of light brown-to-orange discoloration; the pathogen was not consistently recovered from these sections. All other species and the control plants were symptomless. This experiment was repeated with similar results except that inoculated peppers were distinctly stunted compared with control plants. To my knowledge, this is the first report of black root rot caused by T. basicola on lettuce in California. Disease was limited to patches along edges of iceberg lettuce fields; disease incidence in these discrete patches reached as high as 35%. Affected plants continued to grow but remained stunted in relation to unaffected plants and were not harvested. Black root rot of lettuce has been reported in Australia (1); that report also showed that lettuce cultivars vary in susceptibility to T. basicola and isolates from lettuce were highly aggressive on bean but not on many other reported hosts of this pathogen. References: (1) R. G. O'Brien and R. D. Davis. Australas. Plant Pathol. 23:106, 1994. (2) C. V. Subramanian. No. 170 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1968.


2007 ◽  
Vol 119 (4) ◽  
pp. 469-475 ◽  
Author(s):  
Eiko E. Kuramae ◽  
Alexandre L. Buzeto ◽  
Andreia K. Nakatani ◽  
Nilton L. Souza

Plant Disease ◽  
2008 ◽  
Vol 92 (10) ◽  
pp. 1394-1399 ◽  
Author(s):  
Warren E. Copes ◽  
Katherine L. Stevenson

A pictorial key was developed and the relationship between disease severity (S) and incidence (I) was examined to aid in the assessment of black root rot of pansy caused by Thielaviopsis basicola. The key consisted of photographs of root segments that represented nine disease severity levels ranging from 1 to 91%. Pansies that had received different fertility treatments, as part of seven separate experiments, were inoculated with T. basicola. Four weeks after inoculation, roots were washed, and incidence and severity of black root rot were visually assessed using a grid-line-intersect method. Disease incidence ranged from 1.3 to 100%, and severity ranged from 0.1 to 21.4% per plant. Four different mathematical models were compared to quantitatively describe the I-S relationship for the combined data from all seven experiments. Although all models provided an adequate fit, the model that is analogous to the Kono-Sugino equation provided the most reliable estimate of severity over the entire range of disease incidence values. The predictive ability and accuracy of this model across data sets was verified by jackknife and cross-validation techniques. We concluded that incidence of black root rot in pansy can be assessed more objectively and with greater precision than disease severity and can be used to provide reliable estimates of disease severity based on derived regression equations that quantify the I-S relationship for black root rot.


Plant Disease ◽  
2020 ◽  
Author(s):  
Tao Tang ◽  
Fanfan Wang ◽  
Jie Guo ◽  
XiaoLiang Guo ◽  
Yuanyuan Duan ◽  
...  

Ophiopogon japonicus (Linn. f.) is a perennial evergreen in the Liliaceae family that is cultivated in many provinces of China due to its high medicinal and economic value . In April 2019, an unknown root rot disease was observed on the rhizomes of O. japonicus in a commercial production field in Xiangyang City (30.83° N, 112.53° E), Hubei Province. Disease incidence was approximately 10-20%. Symptoms included chlorosis, drooping and rolling of the leaves followed by rapid death of entire plant. Infected roots appeared to be softened, necrotic, and shriveled with reddish fungal growth. Infected tissues were disinfested on surface with 75% ethanol for 30 s and 0.1% HgCl2 for 1 min, rinsed with sterile distilled water, and dried. Small pieces (2 mm × 2 mm) were then excised from disinfested tissue and incubated on potato dextrose agar (PDA) medium at 25 ℃ in the dark. After 3 days of incubation, six isolates with 75% of isolation rate and same colony morphology were sub-cultured and purified by hyphal tip isolation. Purified cultures grew rapidly and media plates (70×70 mm ) were covered with hyphae after 3 to 4 days. Cultures were initially white and became pink or red over 5 days. Microconidia were not observed. Macroconidia were produced from monophialides on branched conidiophores, which were slender, equilaterally curved, and measured 32.5 to 53.5 μm in length and 3.5 to 5.1 μm in width, with three to five septa. All strains were preliminarily identified as Fusarium acuminatum (Eslie and Summerell 2006) on the basis of morphology. To confirm the identity of the pathogen, molecular identification was performed with strain MD1. Following DNA extraction, PCR was performed using the TSINGKE 2×T5 Direct PCR Mix kit. Target areas of amplification were internal transcribed spacer (ITS), RNA polymerase second largest subunit (RPB2) and beta-tubulin gene (TUB2) regions of rDNA, using ITS1,4 (Yin et al. 1990) , RPB2-5f2/7cr (O’Donnell et al. 2010)and Btu-F-F01, Btu-F-R01 primers(Wang et al. 2014), respectively. Nucleotide sequences were deposited in NCBI (GenBank MT525360.1; MW164629; MT588110.1). BLAST analysis of the ITS sequence had 100% similarity to a 517 bp portion of F. acuminatum sequence in GenBank (MK764994.1) ;RPB2 sequence had 100% similarity to a 687 bp portion of F. acuminatum sequence in GenBank (HM068330.1) and TUB2 sequence had 99% similarity to a 964 bp portion of F. acuminatum sequence in GenBank (KT965741.1). A pathogenicity test was performed in laboratory on O. japonicus roots with isolate MD1. Mycelial plugs (5 mm) were excised from the margin of colony cultured for 5 days, and placed on three-years-old tuberous roots covered with wet sterile cotton and kept at 25℃, under 80% relative humidity. Controls were inoculated with non-colonized PDA plugs (5 mm). All treatments had three replicate plants. On incolated plants, white hyphae covered on O. japonicus roots 3 DPI became pink and by 5 DPI, roots had rot symptoms. By comparision, the control plants had no symptoms. The pathogen was reisolated from the inoculated roots and exhibited same morphological characteristics and ITS sequence as those of F. acuminatum. F. acuminatum was reported to cause fruit rot on postharvest pumpkin and Vaccinium corymbosum in China (Li et al. 2019; Wang et al. 2016).To our knowledge, this is the first report of root rot caused by F. acuminatum on O. japonicus in China.


Plant Disease ◽  
2001 ◽  
Vol 85 (11) ◽  
pp. 1200-1205 ◽  
Author(s):  
A. Priyatmojo ◽  
Y. Yotani ◽  
K. Hattori ◽  
K. Kageyama ◽  
M. Hyakumachi

Root and stem rot of miniature rose (Rosa hybrida L.) was observed in commercial glasshouse-grown roses in Gifu prefecture, Japan, during the summer and fall of 1997 and 1998. One hundred and fifty-three isolates of Rhizoctonia spp. were obtained from infected roots and stems. Of the 153 isolates, 9 had binucleate and 144 had multinucleate vegetative hyphal cells. Binucleate Rhizoctonia failed to anastomose with tester isolates of anastomosis groups (AG)-A through -S (not including AG-J and AG-M). Of 144 isolates identified as R. solani, 83.3% were AG 2-2 IIIB and 16.7% were AG 4 HG-I. Five isolates from each group caused severe rot and mortality on cuttings during rooting. Pathogenicity of Rhizoctonia spp. varied on three different ages of miniature roses cv. Silk. Isolates of AG 4 HG-I caused root and stem rot and mortality on 15-, 25-, and 40-day-old plants, whereas isolates of AG-2-2 IIIB caused root and stem rot and mortality on 15- and 25-day-old plants, but light root rot on 40-day-old plants. Isolates of binucleate Rhizoctonia caused root and stem rot and mortality only on 15-day-old plants.


Plant Disease ◽  
2011 ◽  
Vol 95 (9) ◽  
pp. 1194-1194 ◽  
Author(s):  
G. Polizzi ◽  
D. Aiello ◽  
V. Guarnaccia ◽  
A. Panebianco ◽  
P. T. Formica

The genus Passiflora (Passifloraceae family) contains more than 500 species and several hybrids. In Italy, some of these species and hybrids are grown as ornamental evergreen vines or shrubs. During August and September 2010, a crown and root rot was observed in a stock of approximately 6,000 potted 2-year-old plants of Passiflora mollissima (Kunth) Bailey, commonly known as the banana passionflower, in a nursery located in eastern Sicily (southern Italy). Disease incidence was approximately 20%. Disease symptoms consisted of water-soaked lesions at the crown and a root rot. Successively, older crown lesions turned light brown to brown and expanded to girdle the stem. As crown and root rot progressed, basal leaves turned yellow and gradually became necrotic and infected plants wilted and died. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently isolated from crown lesions and brown decaying roots when plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 μg/ml. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Mycelium was branched at right angles with a septum near the branch with a slight constriction at the branch base. Hyphal cells removed from 10 representative cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (4). Pairings were made with tester strains of AG-1, AG-2, AG-3, AG-4, AG-5, AG-6, and AG-11. Anastomosis was observed only with tester isolates of AG-4 (3). Pathogenicity tests were performed on container-grown, healthy, 3-month-old cuttings. Twenty plants of P. mollissima were inoculated near the base of the stem with five 1-cm2 PDA plugs from 5-day-old mycelial plugs obtained from two representative cultures. The same number of plants served as uninoculated controls. Plants were maintained at 25°C and 95% relative humidity with a 12-h fluorescent light/dark regimen. Wilt symptoms due to crown and root rot, identical to ones observed in the nursery, appeared 7 to 8 days after inoculation with either of the two isolates and all plants died within 20 days. No disease was observed on control plants. R. solani AG-4 was reisolated from symptomatic tissues and identified as previously described, confirming its pathogenicity. Damping-off or crown and root rot due to R. solani were previously detected on P. edulis in Brazil, Africa, India, Oceania, and Australia (2). To our knowledge, this is the first report of R. solani causing crown and root rot on P. mollissima. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) J. L. Bezerra and M. L. Oliveira. Fitopathol. Brasil. 9:273, 1984. (3) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.


Author(s):  
Mahfouz M. M. Abd-Elgawad ◽  
Ibrahim E. Elshahawy ◽  
Farid Abd-El-Kareem

Abstract Background Strawberry (Fragaria ananassa Duch.) is an economically important crop in Egypt. Yet complex black root rot disease of strawberry caused by Fusarium solani, Rhizoctonia solani, and Pythium sp. can cause considerable yield losses. Therefore, this study aimed at evaluating different aspects of soil solarization against this disease. Such an evaluation would better be viewed in the context of other beneficial effects of soil solarization on nematodes and weeds. Materials/methods Growth agar disks, growth suspension, and resting stages of strawberry black root rot fungi were evaluated at different temperatures and exposure times using digital hot water bath. Cloth bags artificially infested with single fungal species were buried into the soil before soil solarization at soil depths of 1–10, 11–20, and 21–30 cm at three spots of each plot for each of the abovementioned fungi for 3, 6, or 9 weeks. The disease incidence and severity in solarized and un-solarized soil was compared with the application of the fungicide Actamyl. Effects of soil solarization on nematodes and weeds were also consulted. Results The lethal temperature to F. solani, Pythium sp., and R. solani was 58, 58, and 56 °C, respectively when exposure time was 1 min. Chlamydospores were killed at 62 °C while sclerotia were killed at 58 °C in hot water for 1 min. Maximum soil temperature in solarized soil was raised by 15, 14, and 12 °C at depths of 1–10, 11–20, and 21–30 cm as compared with non-solarized soil. Solarization for 3, 6, and 9 weeks significantly reduced the disease incidence and severity and increased the strawberry yield. Complete reduction in total count of all tested fungi was obtained after 9 weeks at all tested depths. A review of collective soil pest and pathogen control via solarization documented its beneficial application. Conclusion The study may exploit hot months in Egypt for soil solarization against the serious root rot disease either singly or in an integrated pest management program.


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