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Article ; Online: Positive selection and functional diversification of transcription factor Cmr1 homologs in Alternaria.

Qiu, Chaodong / Liu, Zhenyu

2024 Volume 108, Issue 1, Page(s) 133

Abstract: Transcription factor Cmr1 (Colletotrichum melanin regulation 1) and its homologs in several plant fungal pathogens are the regulators of the 1,8-dihydroxynaphthalene (DHN)-melanin biosynthesis pathway and have evolved functional diversification in ... ...

Abstract	Transcription factor Cmr1 (Colletotrichum melanin regulation 1) and its homologs in several plant fungal pathogens are the regulators of the 1,8-dihydroxynaphthalene (DHN)-melanin biosynthesis pathway and have evolved functional diversification in morphology and pathogenicity. The fungal genus Alternaria comprises the group of "black fungi" that are rich in DHN-melanin in the primary cell wall and septa of the conidia. Some Alternaria species cause many economically important plant diseases worldwide. However, the evolution and function of Cmr1 homologs in Alternaria remain poorly understood. Here, we identified a total of forty-two Cmr1 homologs from forty-two Alternaria spp. and all contained one additional diverse fungal specific transcription factor motif. Phylogenetic analysis indicated the division of these homologs into five major clades and three branches. Dated phylogeny showed the A and D clades diverged latest and earliest, respectively. Molecular evolutionary analyses revealed that three amino acid sites of Cmr1 homologs in Alternaria were the targets of positive selection. Asmr1, the homolog of Cmr1 in the potato early blight pathogen, Alternaria solani was amplified and displayed the sequence conservation at the amino acid level in different A. solani isolates. Asmr1 was further confirmed to have the transcriptional activation activity and was upregulated during the early stage of potato infection. Deletion of asmr1 led to the decreased melanin content and pathogenicity, deformed conidial morphology, and responses to cell wall and fungicide stresses in A. solani. These results suggest positive selection and functional divergence have played a role in the evolution of Cmr1 homologs in Alternaria. KEY POINTS: • Cmr1 homologs were under positive selection in Alternaria species • Asmr1 is a functional transcription factor, involved in spore development, melanin biosynthesis, pathogenicity, and responses to cell wall and fungicide stresses in A. solani • Cmr1 might be used as a potential taxonomic marker of the genus Alternaria.
MeSH term(s)	Transcription Factors/genetics ; Transcription Factors/metabolism ; Alternaria/genetics ; Alternaria/metabolism ; Melanins/metabolism ; Fungicides, Industrial/metabolism ; Phylogeny ; Naphthols
Chemical Substances	Transcription Factors ; 1,8-dihydroxynaphthalene melanin ; Melanins ; Fungicides, Industrial ; Naphthols
Language	English
Publishing date	2024-01-15
Publishing country	Germany
Document type	Journal Article
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-023-12893-7
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Article ; Online: Multifunctionality of AsCFEM6 and AsCFEM12 effectors from the potato early blight pathogen Alternaria solani.

Qiu, Chaodong / Halterman, Dennis / Zhang, Huajian / Liu, Zhenyu

International journal of biological macromolecules

2023 Volume 257, Issue Pt 1, Page(s) 128575

Abstract: Plant pathogens secrete fungal-specific common in several fungal extracellular membrane (CFEM) effectors to manipulate host immunity and contribute to their virulence. Little is known about effectors and their functions in Alternaria solani, the ... ...

Abstract	Plant pathogens secrete fungal-specific common in several fungal extracellular membrane (CFEM) effectors to manipulate host immunity and contribute to their virulence. Little is known about effectors and their functions in Alternaria solani, the necrotrophic fungal pathogen causing potato early blight. To identify candidate CFEM effector genes, we mined A. solani genome databases. This led to the identification of 12 genes encoding CFEM proteins (termed AsCFEM1-AsCFEM12) and 6 of them were confirmed to be putative secreted effectors. In planta expression revealed that AsCFEM6 and AsCFEM12 have elicitor function that triggers plant defense response including cell death in different botanical families. Targeted gene disruption of AsCFEM6 and AsCFEM12 resulted in a change in spore development, significant reduction of virulence on potato and eggplant susceptible cultivars, increased resistance to fungicide stress, variation in iron acquisition and utilization, and the involvement in 1,8-dihydroxynaphthalene (DHN) melanin biosynthesis pathway. Using maximum likelihood method, we found that positive selection likely caused the polymorphism within AsCFEM6 and AsCFEM12 homologs in different Alternaria spp. Site-directed mutagenesis analysis indicated that positive selection sites within their CFEM domains are required for cell death induction in Nicotiana benthamiana and are critical for response to abiotic stress in yeast. These results demonstrate that AsCFEM effectors possess additional functions beyond their roles in host plant immune response and pathogen virulence.
MeSH term(s)	Alternaria/physiology ; Genes, Fungal ; Plant Diseases/microbiology ; Solanum tuberosum/genetics ; Solanum tuberosum/microbiology ; Virulence/genetics
Language	English
Publishing date	2023-12-02
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	282732-3
ISSN	1879-0003 ; 0141-8130
ISSN (online)	1879-0003
ISSN	0141-8130
DOI	10.1016/j.ijbiomac.2023.128575
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Article: Nigrospora oryzae Causing Leaf Spot on Asiatic Dayflower in Chongqing, China

Qiu, Chaodong / Zhu, Wenhui / Niu, Taotao / Liu, Zhenyu

Plant disease. 2022 Feb. 28, v. 106, no. 2

2022

Abstract: Asiatic dayflower (Commelina communis L.) is an annual herbaceous weed that is distributed throughout China. A foliar disease on Asiatic dayflowers was discovered in one farm field in Dianjiang County, Chongqing, China (30°3′22″ N, 107°18′5″ E) in summer ...

Abstract	Asiatic dayflower (Commelina communis L.) is an annual herbaceous weed that is distributed throughout China. A foliar disease on Asiatic dayflowers was discovered in one farm field in Dianjiang County, Chongqing, China (30°3′22″ N, 107°18′5″ E) in summer 2019. The disease incidence was observed on about 10% (13/127) of the plants. Symptoms appeared as round, tan lesions 2 to 5 mm in diameter that occurred randomly and irregularly on the whole leaves. The centers of lesions become grayish white with reddish borders as the disease progressed. The leaves with typical symptoms were detached and wiped with 70% ethanol for surface disinfestation before isolating the causal agent. Subsequently, three pieces (3 to 4 mm²) of tissue were taken from the margin of the leaf lesion, disinfested in 1.5% NaClO for 1 min, rinsed three times in sterilized distilled water, and placed onto potato dextrose agar (PDA) medium containing 50 μg/ml each of kanamycin and ampicillin. A fungus was exclusively and consistently isolated from the disinfested leaf lesion sections. The colonies on PDA grew rapidly and covered the entire Petri dish within 5 days at 28°C. Colonies were at first grayish white, cotton wool-like, round, with abundant aerial mycelium, and later turned black as conidia were produced. The abundant conidia formed on PDA were initially yellow brown and gradually became black, oblate to ellipsoidal, smooth, single-celled, and ranged in size from 4 to 10 × 3.5 to 9 μm. They were borne on a colorless, hyaline, and inverted flask-shaped cell at the tip of each conidiophore. The morphology characteristics were consistent with those of Nigrospora spp. (Wang et al. 2017). Genomic DNA was extracted from one representative isolate NDJ0819. The amplification and sequencing of the gene fragments including the internal transcribed space (ITS) region of ribosomal DNA and beta-tubulin were performed using the primers ITS1/ITS4 (White et al. 1990) and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. Fragments of 536 bp for ITS and 408 bp for beta-tubulin were obtained. A phylogram of the combined ITS and beta-tubulin sequences reconstructed using the maximum likelihood bootstrapping method implemented in the software MEGA version 7.0 (Kumar et al. 2016) indicated that isolate NDJ0819 clustered with Nigrospora oryzae. Both ITS and beta-tubulin sequences were deposited into GenBank (accession nos. MT140353 and MT157509, respectively). Pathogenicity test was performed by rub-inoculating needle-wounded leaves of three 4-week-old Asiatic dayflowers with a spore suspension (2.6 × 10⁶ conidia/ml) of NDJ0819 prepared in water containing 0.05% Tween-20, and holding plants at 28°C in the growth chamber. The pathogenicity test was repeated twice. Brown, round lesions developed on leaves inoculated with spores at 15 days postinoculation. However, the centers of the lesion did not become grayish white, compared with those of lesions seen in naturally infected leaves. No symptoms developed on leaves inoculated with sterilized distilled water. N. oryzae was reisolated from the lesions. All results described above indicated that N. oryzae was responsible for the leaf spot of Asiatic dayflower. To our knowledge, this is the first report of N. oryzae causing leaf spot on Asiatic dayflower in China. Research into the potential use of N. oryzae as a candidate biological agent against the weed is worth being initiated.
Keywords	Commelina communis ; Khuskia oryzae ; Nigrospora ; ampicillin ; computer software ; conidia ; cotton ; culture media ; disease incidence ; disinfestation ; ethanol ; farms ; foliar diseases ; fungi ; genes ; growth chambers ; kanamycin ; leaf spot ; leaves ; mycelium ; pathogenicity ; polysorbates ; ribosomal DNA ; statistical analysis ; summer ; tubulin ; weeds ; China
Language	English
Dates of publication	2022-0228
Publishing place	The American Phytopathological Society
Document type	Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-06-21-1271-PDN
Database	NAL-Catalogue (AGRICOLA)

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Article: Genome Resource of American Ginseng Black Spot Pathogen

Qiu, Chaodong / Wang, Weiquan / Liu, Zhenyu

Plant disease

2022 Volume 106, Issue 3, Page(s) 1020–1022

MeSH term(s)	Alternaria/genetics ; Panax/genetics
Language	English
Publishing date	2022-03-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-05-21-0895-A
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: First Report of Alternaria spp. Causing Leaf Spot on Sweet Viburnum in China

Qiu, Chaodong / Zhang, Yingying / Liu, Zhenyu

Plant disease. 2021 Aug. 20, v. 105, no. 8

2021

Abstract: Sweet viburnum (Viburnum odoratissimum [L.] Ker Gawl) is an evergreen shrub mainly cultivated along roadsides in urban landscapes and in parks and residential areas. A foliar disease occurred on ∼40% of sweet viburnum plants near Anhui Grand Theater, ... ...

Abstract	Sweet viburnum (Viburnum odoratissimum [L.] Ker Gawl) is an evergreen shrub mainly cultivated along roadsides in urban landscapes and in parks and residential areas. A foliar disease occurred on ∼40% of sweet viburnum plants near Anhui Grand Theater, Anhui Province, China, in June 2019. Early symptoms appeared as small brown spots 2 to 3 mm in length on the leaves. Spots developed on the upper, middle, and lower leaves; however, upper leaves were more severely affected. As the disease developed, spots enlarged and became rectangular or oval, brown to dark brown, and their centers became ashen gray. In later stages, diseased leaves wilted. Diseased leaves were surface disinfested, and three 2- to 3-mm² sections were cut from lesion margins. Sections were placed in 1.5% NaClO for 2 min, submerged in three changes of sterilized distilled water (SDW) for 1 min each, placed onto PDA amended with 50 μg/ml of ampicillin and kanamycin, and incubated at 25°C for 3 days. The mycelium from the leading edge of colonies growing from the tissue was subcultured onto a PDA plate for 3 days, followed by spore induction (Simmons 2007) and single-spore isolation to obtain a pure culture. Colonies of single-spore isolate HF0719 were rounded and grayish white with dense aerial mycelium viewed from above and dark brown viewed from below. On potato carrot agar, conidiophores were branched or occasionally unbranched. On branched conidiophores, conidia were in dwarf tree-like branched chains of 2 to 5 conidia. On unbranched conidiophores, conidia were simple or in chains of 2 to 8 conidia. Conidia were light brown or dark brown, ovoid, ellipsoidal to fusiform, and 7 to 26.5 × 4.5 to 11 μm (average 16 × 7 µm, n = 500 spore observations), with one beak and 1 to 7 transverse, 0 to 3 longitudinal, and 0 to 3 oblique septa. Beaks were (1.5 to) 2 to 10 (to 16) μm long. Based on cultural and morphological characteristics, HF0719 was identified as Alternaria spp. (Simmons 2007). For molecular identification, total genomic DNA was isolated from mycelia collected from 7-day-old colonies of HF0719 using the fungal genomic DNA extraction kit (Solarbio, Beijing, China). Fragments of five genes, including those encoding GAPDH (gpd), plasma membrane ATPase, actin, calmodulin, and the Alternaria major allergen (Alt a1) regions of HF0719 were amplified and sequenced using primer pairs gpd1/gpd2 (Berbee et al. 1999), ATPDF1/ATPDR1, ACTDF1/ACTDR1, CALDF1/CALDR1 (Lawrence et al. 2013), and Alt-for/Alt-rev (Hong et al. 2005), respectively. The obtained nucleotide sequences were deposited in GenBank: gpd, MT614365; ATPase, MT614364; actin, MT614363; calmodulin, MN706159; and Alt a1, MN304720. A phylogenetic tree using a maximum likelihood bootstrapping method based on the five-gene combined dataset (gpd, ATPase, actin, calmodulin, and Alt a1) of HF0719 and standard strains of 120 Alternaria species (Lawrence et al. 2013) was constructed. HF0719 formed a separate branch. Based on morphology and phylogenetic pattern, HF0719 was identified as Alternaria spp. Pathogenicity was tested by rubbing 32 healthy leaves of six 5-year-old sweet viburnum plants with a cotton swab dipped in spore suspension containing 2.6 × 10⁶ spores/ml, following leaf surface disinfection with 70% ethanol in the open field. SDW was used as a control. Average air temperature was ∼28°C. Eleven days after inoculation, 100% of inoculated leaves showed symptoms identical to those in the field. Control leaves were symptomless. The experiment was done three times. The reisolated pathogen from the leaf lesion had the same morphological and molecular characteristics as HF0719, satisfying Koch’s postulates. Alternaria has been reported to cause leaf spot on sweet viburnum in Florida, U.S.A. (Alfieri et al. 1984). This is the first report of Alternaria spp. causing leaf spot on sweet viburnum in China. Our findings will contribute to monitoring and adopting strategies to manage leaf spot disease on sweet viburnum.
Keywords	Alternaria ; DNA ; Viburnum odoratissimum ; actin ; adenosinetriphosphatase ; air temperature ; allergens ; ampicillin ; beak ; calmodulin ; conidia ; conidiophores ; cotton ; culture media ; data collection ; disinfection ; ethanol ; foliar diseases ; fungi ; kanamycin ; leaf spot ; leaves ; mycelium ; pathogenicity ; pathogens ; phylogeny ; plasma membrane ; shrubs ; statistical analysis ; China ; Florida
Language	English
Dates of publication	2021-0820
Publishing place	The American Phytopathological Society
Document type	Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-06-20-1407-PDN
Database	NAL-Catalogue (AGRICOLA)

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Article: First Report of Leaf Spot of Weigela florida Caused by Epicoccum layuense in China

Tian, Yue / Zhang, Yingying / Qiu, Chaodong / Liu, Zhenyu

Plant disease. 2021 Aug. 20, v. 105, no. 8

2021

Abstract: Weigela florida (Bunge) A. DC. is a dense, rounded, deciduous shrub commonly planted in landscapes. It is also used in Chinese medicine to treat sore throat, erysipelas, cold, and fever (Zheng et al. 2019). In May 2019, leaf spots were observed on ... ...

Abstract	Weigela florida (Bunge) A. DC. is a dense, rounded, deciduous shrub commonly planted in landscapes. It is also used in Chinese medicine to treat sore throat, erysipelas, cold, and fever (Zheng et al. 2019). In May 2019, leaf spots were observed on approximately 50% of W. florida plants grown in the Wisdom Plaza Park of Anhui Agricultural University in Hefei, Anhui Province, China. Leaf spots began as small light brown and irregular lesions, enlarged, turned reddish brown, coalesced to form large blighted areas, and eventually covered the entire leaf surface. Five pieces of tissues were removed from the lesion margins of each diseased leaf (five leaves from five different plants), chopped into several 3- to 4-mm² pieces, disinfected with 1.5% NaOCl for 2 min, rinsed three times with sterile distilled water for 1 min, plated onto potato dextrose agar (PDA) medium containing 50 μg/ml of ampicillin and kanamycin, and incubated at 25°C with a 12-h photoperiod for 5 days. One segment of the fungal growth from the growing edge of the colony was transferred onto a fresh PDA plate for purification and incubated under the same conditions for another 5 days. The colony morphology of one representative isolate (AAU0519) was characterized by a pale orange cushion in the center surrounded by irregular pink margin, diffusing red-orange pigments into the PDA medium. Isolate AAU0519 was cultured on PDA medium for 7 days at 25°C in the dark to induce sporulation. The produced conidia were globose, subglobose to pyriform, golden brown to brown, and with a diameter of 7.7 to 23.8 μm. Both cultural and morphological characteristics suggested that isolate AAU0519 was an Epicoccum species, according to the description by Chen et al. (2017). Amplification and sequencing of the internal transcribed spacer (ITS), beta-tubulin, and 28S large subunit ribosomal RNA (LSU) gene fragments from the extracted genomic DNA of AAU0519 were performed using primer sets ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass and Donaldson 1995), and LSU1Fd/LR5 (Crous et al. 2009; Vilgalys and Hester 1990), respectively. A phylogenetic tree was constructed by the maximum-likelihood method with 1,000 bootstrap replications based on the concatenated ITS, beta-tubulin, and LSU sequences from isolate AAU0519 and representative strains of 22 species of the genus Epicoccum (Chen et al. 2017). Isolate AAU0519 clustered with ex-holotype CGMCC 3.18362 of Epicoccum layuense Q. Chen, Crous & L. Cai (Chen et al. 2017). All obtained sequences were deposited into GenBank under accession numbers MK983497 (ITS), MN328723 (beta-tubulin), and MN328724 (LSU). A pathogenicity test was conducted on leaves of five 3-year-old W. florida cultivar ‘Red Prince’ planted in the field (five leaves for each treatment and control per plant) by spraying 30 ml of a spore suspension (10⁶ spores/ml) of isolate AAU0519 as the treatment or sterilized distilled water as the control. Before the inoculation, the leaves were disinfected with 70% ethanol. After inoculation, the leaves were wrapped with a plastic bag to keep high relative humidity. The average air temperature was about 28°C during the period of the pathogenicity test. The experiment was repeated once. Ten days after inoculation, the fungal-inoculated leaves developed light brown lesions resembling those of naturally infected leaves, and control leaves did not develop any symptoms. E. layuense was recovered from leaf lesions, and its identity was confirmed by morphological and sequence analyses as described above. To our knowledge, E. layuense has been previously reported as a pathogen of Perilla sp. (Chen et al. 2017), oat (Avena sativa) (Chen et al. 2020a), and tea (Camellia sinensis) plants (Chen et al. 2020b), but this is the first report of E. layuense causing leaf spot on W. florida in China. This pathogen could pose a threat to the ornamental value of W. florida plants. Thus, it is necessary to adopt effective management strategies against leaf spot on W. florida.
Keywords	Avena sativa ; Camellia sinensis ; Epicoccum ; Oriental traditional medicine ; Perilla ; Weigela florida ; agricultural colleges ; air temperature ; ampicillin ; cold ; conidia ; cultivars ; culture media ; erysipelas ; ethanol ; fever ; fungal growth ; genes ; internal transcribed spacers ; kanamycin ; leaf spot ; leaves ; oats ; ornamental value ; pathogenicity ; pathogens ; pharyngitis ; phylogeny ; relative humidity ; ribosomal RNA ; shrubs ; sporulation ; statistical analysis ; tea ; tubulin ; China
Language	English
Dates of publication	2021-0820
Publishing place	The American Phytopathological Society
Document type	Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-07-20-1498-PDN
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Article: First Report of Leaf Spot of

Tian, Yue / Zhang, Yingying / Qiu, Chaodong / Liu, Zhenyu

Plant disease

2021

Abstract: ... Weigela ... ...

Abstract	Weigela florida
Language	English
Publishing date	2021-03-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-07-20-1498-PDN
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Article: First Report of

Qiu, Chaodong / Zhang, Yingying / Liu, Zhenyu

Plant disease

2021

Abstract: Sweet viburnum [ ...

Abstract	Sweet viburnum [
Language	English
Publishing date	2021-02-19
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-06-20-1407-PDN
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Article: Nigrospora oryzae

Qiu, Chaodong / Zhu, Wenhui / Niu, Taotao / Liu, Zhenyu

Plant disease

2021

Abstract: Asiatic dayflower ( ...

Abstract	Asiatic dayflower (
Language	English
Publishing date	2021-08-31
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-06-21-1271-PDN
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Article: Nigrospora sphaerica causing leaf spot in a new host, Eclipta prostrata (False Daisy), in China

Qiu, Chaodong / Liu, Chen / Niu, Taotao / Zhu, Wenhui / Liu, Zhenyu

Phytopathologische Zeitschrift. 2022 Apr., v. 170, no. 4

2022

Abstract: Leaf spot symptom was observed in false daisy [Eclipta prostrata (L.) L., syn. Eclipta alba (L.) Hassk.] in Dianjiang County, Chongqing, China, in August 2019. The isolated and purified fungus CQLC820 was confirmed as Nigrospora sphaerica based on ... ...

Abstract	Leaf spot symptom was observed in false daisy [Eclipta prostrata (L.) L., syn. Eclipta alba (L.) Hassk.] in Dianjiang County, Chongqing, China, in August 2019. The isolated and purified fungus CQLC820 was confirmed as Nigrospora sphaerica based on cultural and morphological features and phylogram of combined sequences of gene fragments of the internal transcribed spacer (ITS) region of ribosomal DNA and beta‐tubulin. Pathogenicity test revealed that isolate CQLC820 caused leaf spot lesions on false daisy, identical to those occurred naturally in the field. N. sphaerica was re‐isolated from the lesions and identified through morphological and sequence analysis. The causal agent of leaf spot of false daisy was identified as N. sphaerica. This is the first time to identify false daisy as a new host of N. sphaerica not only in China, but also in the world. There is a potential concern that N. sphaerica might have a threat on production of false daisy, a traditional herbal medicine with a long history of uses for treating haemorrhagic diseases in Asia and South America.
Keywords	Eclipta prostrata ; Nigrospora ; fungi ; genes ; herbal medicines ; internal transcribed spacers ; leaf spot ; pathogenicity ; ribosomal DNA ; sequence analysis ; tubulin ; China ; South America
Language	English
Dates of publication	2022-04
Size	p. 242-246.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	633487-8
ISSN	0931-1785
ISSN	0931-1785
DOI	10.1111/jph.13075
Database	NAL-Catalogue (AGRICOLA)

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