Use of single-protoplast isolates in the study of the mating phenomena of Rhizoctonia solani (Thanatephorus cucumeris) AG-1 IC and IA

Qu, Ping; Aratani, Akiko; Syoji, Takako; Toda, Takeshi; Kubota, Mayumi; Hyakumachi, Mitsuro

doi:10.1007/s10267-007-0400-6

Cited by 7 publications

(7 citation statements)

References 23 publications

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“…Consequently, most investigations have focused on haploid protoplasts rather than single basidiospores. Such studies have confirmed that AG 1‐IA can form both homokaryotic and heterokaryotic progeny 12 . In addition, changes to the protoplast‐releasing procedure have been found to result in protoplasts that contain different numbers of nuclei, making them genetically different from their parental isolates 6 …”

Section: Introductionmentioning

confidence: 82%

“…Ten mycelial plugs (5 mm in diameter) were then cut from the intersection of the two colonies and used to inoculate Potato Dextrose Broth (PDB). After 24 h of dark incubation at 25°C with shaking (120 rpm), the mycelium was removed with forceps, washed in an osmotic medium (0.98 m MgSO 4 , 8.4 m m Na 2 HPO 4 , 1.6 m m NaH 2 PO 4 ), 44 and ground with a sterile pestle and mortar before being transferred to a 250‐ml flask containing 100 ml CM medium (5 g L −1 glucose, 5 g L −1 malt extract, 5 g L −1 yeast extract), 12 and dark‐incubated at 25°C with shaking (120 rpm) for 24 h. The mycelium was then collected by centrifugation at 290 g for 10 min. After the supernatant had been removed, the mycelium was washed in 10 ml osmotic medium and centrifuged as before.…”

Section: Methodsmentioning

confidence: 99%

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Heterokaryotic state of a point mutation (H249Y) in SDHB protein drives the evolution of thifluzamide resistance in Rhizoctonia solani

Miao

et al. 2020

Pest Management Science

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BACKGROUND Sheath blight, caused by Rhizoctonia solani, can be effectively controlled by application of the succinate dehydrogenase inhibitor thifluzamide. Although the risk of resistance to thifluzamide in R. solani had been reported, the thifluzamide‐resistance mechanism and the evolution of thifluzamide‐resistance in R. solani have not been investigated in detail. RESULTS No differences were found between the sequences of proteins SDHA, SDHC, and SDHD in thifluzamide‐sensitive isolates and thifluzamide‐resistant mutants, but a single point mutation H249Y was found in SDHB. Two different types of thifluzamide‐resistant R. solani mutants were characterized: homokaryotic, carrying only the resistance allele; and heterokaryotic, retaining the wild‐type allele in addition to the resistance allele. The resistance level differed according to the nuclear composition at codon 249 in the sdhB gene. Molecular docking results suggested that the point mutation (H249Y) might significantly alter the affinity of thifluzamide and SDHB protein. Heterokaryotic mutants were able to evolve into a homokaryon when repeatedly cultured on agar media or rice plants in the presence of thifluzamide, but thifluzamide treatment had no effect on the genotypes of homokaryotic mutants or sensitive isolates. CONCLUSION This study showed that H249Y in SDHB protein could cause thifluzamide resistance in R. solani. Fungicide application could promote heterokaryotic mutants to evolve into a homokaryon. © 2020 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 82%

Section: Methodsmentioning

confidence: 99%

Heterokaryotic state of a point mutation (H249Y) in SDHB protein drives the evolution of thifluzamide resistance in Rhizoctonia solani

Miao

et al. 2020

Pest Management Science

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“…When one of the homokaryons was paired separately with the other three homokaryons, none of the parings showed tuft formation, indicating that all of them belonged to the same type of homokaryon. Since R. solani AG1-IC has been reported to produce two types of homokaryons (Ou et al [ 2008 ]), this type of homokaryon was designated as A, while the other not yet found homokaryon type was designated as B. The heterokaryon was designated as AB.…”

Section: Methodsmentioning

confidence: 99%

“…In R. solani , homokaryotic isolates are normally derived from single basidiospores (Whitney and Parmeter [ 1963 ]) or protoplasts (Phillips [ 1993 ]). Since R. solani AG1-IC is heterothallic (Qu et al [ 2008 ]) and our numerous attempts to induce basidiospore formation of isolate CH-1 also were not successful, protoplasts were used for preparation of homokaryons. In the process of homokaryon preparation, it was found that colonies derived from protoplasts of isolate CH-1 consisted of heterokaryon and one type of homokaryon only.…”

Section: Introductionmentioning

confidence: 99%

A new variant produced by Rhizoctonia solani AG1-IC isolate CH-1 with a new type of nuclei

Tsai

2014

Bot Stud

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BackgroundIsolate CH-1 of Rhizoctonia solani Kühn was commonly used in our studies of the pathogenicity and genetics of this pathogen. During the preparation of homokaryons through protoplast regeneration and tuft formation, a defective homokaryon was detected and a new variant was obtained.ResultsWhen tuft formation was used to identify the karyotic nature of single protoplast regenerants (SPRs) of Rhizoctonia solani AG1-IC isolate CH-1, one homokaryon type designated as A type and the parental heterokaryon designated as AB type were obtained. The homokaryon B type was not found. Various approaches were used to obtain SPRs, including from fast or slow growing protoplast regenerants, and from regenerants of protoplasts released from mycelia grown in different nutrient broths or at different temperatures. Without exception, all these SPRs were either homokaryon A or heterokaryon AB. Moreover, the SPRs obtained from different generations of SPRs, and from different generations of SPRs treated with lytic enzymes 3 to 4 times also were invariably either homokaryon A or heterokaryon AB. When single hyphal isolates were obtained from the tuft resulting from the pairing between homokaryon A and heterokaryon AB, only the heterokaryon and a variant were obtained. The variant did not form tuft when paired with parental heterokaryon AB or homokaryon A. Its protoplast regenerants gave rise to heterokaryon AB, homokaryon A and the variant, indicating that it is a new kind of heterokaryon.ConclusionInability to obtain homokaryon B despite numerous attempts suggests that the B type nuclei are probably defective and are dependent on A type nuclei for their multiplication. This is the first report of a heterokaryotic R. solani strain carrying a defective type of nuclei. A new variant which is a new kind of heterokaryon was obtained from the tuft resulting from the paring between the homokaryon A and the parental heterokaryon AB.Electronic supplementary materialThe online version of this article (doi:10.1186/s40529-014-0069-z) contains supplementary material, which is available to authorized users.

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“…Regeneration of protoplasts of R. solani was usually carried out on agar media [15,16,19,26]. When tested in liquid media, Robin-…”

Section: Figurementioning

confidence: 99%

Factors affecting protoplast formation by Rhizoctonia solani

Liu

Lin

2010

New Biotechnology

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Novozym 234 was the most frequently used enzyme for production of Rhizoctonia solani protoplasts. Since manufacture of this enzyme was discontinued in the late 1990s, a new procedure was developed by testing lytic enzymes from Sigma and by examining factors affecting protoplast formation. The combination of 20 mg/mL Driselase and 10mg/mL lysing enzyme was effective in releasing protoplasts from R. solani. The optimal condition for enzyme treatment of mycelium was incubation at 37 degrees C for 15 min followed by 34 degrees C for 105 min. The amount of protoplasts produced was positively correlated with growth rate and negatively correlated with mycelial density. Under favorable conditions, R. solani mycelia released 1.68 x 10(6) protoplasts/mL that is comparable with that produced with Novozym 234. Among various media tested, the best solid medium for protoplast regeneration was 1% V-8 juice agar, while the best liquid medium was 10% potato dextrose broth.

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Use of single-protoplast isolates in the study of the mating phenomena of Rhizoctonia solani (Thanatephorus cucumeris) AG-1 IC and IA

Cited by 7 publications

References 23 publications

Heterokaryotic state of a point mutation (H249Y) in SDHB protein drives the evolution of thifluzamide resistance in Rhizoctonia solani

Heterokaryotic state of a point mutation (H249Y) in SDHB protein drives the evolution of thifluzamide resistance in Rhizoctonia solani

A new variant produced by Rhizoctonia solani AG1-IC isolate CH-1 with a new type of nuclei

Factors affecting protoplast formation by Rhizoctonia solani

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