First Report of Colletotrichum fructicola Causing Anthracnose on Cherry (Prunus avium) in China

Abstract

Cherry (Prunus avium) has become an important economical fruit in China. In October 2020, a leaf spot disease was found on cherry in the orchard of Taizhou Academy of Agriculture Sciences, Zhejiang, China. The symptoms appeared as small, water-soaked spots on the leaves, which later became larger, dark brown, and necrotic lesions of 1 cm to 3 cm in width, 4 cm to 8 cm in length. Disease incidences of approximately 60% of the leaves were observed by sampling five locations. To isolate the causing agent, small fragments from five target symptomatic leaves were surface-sterilized with 1.0% sodium hypochlorite solution for 1 min and then rinsed three times with sterilized water. Afterwards the leaf fragments were air-dried, plated onto potato dextrose agar (PDA) medium, and incubated at 25 ℃ in the dark for 2 days. The pure cultures were obtained by transferring hyphal plug of 2 mm in diameter onto PDA, which followed single spore isolation. The colony morphology showed light to dark gray, cottony mycelium, with the underside of the culture became brownish after 7 days. Conidia (n = 28) were hyaline, smooth-walled, cylindrical, aseptate, broadly rounded ends, and average size around 3.84 × 12.82 μm (2.99 to 4.87 × 10.27 to 15.68 μm). Appressoria (n = 27) were mostly brown, ovoid and slightly irregular in shape, and average size around 8.04 × 9.68 μm (6.29 to 9.67 × 9.32 to 12.06 μm). Perithecia average size is 106.25 μm, textura angularis, thick-walled. Asci 26.35–49.18 × 5.00-12.03 μm (average size 37.44 × 7.80 μm, n = 17), unitunicate, thin-walled, clavate or cymbiform. Ascospores 13.69–20.93 × 3.86-6.69 μm (average size 16.00 × 5.42 μm, n = 30), one-celled, hyaline, one or two large guttulate at the centre, slightly rounded ends. The morphological characteristics matched well with previous descriptions of Colletotrichum species of C. gloeosporioides species complex, including C. fructicola (Prihastuti et al. 2009; Fu et al. 2019). The identity of two representative isolates (cf2-3 and cf4-4) from different leaves was confirmed by means of multi-locus gene sequencing. To this end, genomic DNA was extracted by the Plant Direct PCR kit (Vazyme Biotech Co., Ltd, China). Molecular identification was conducted by sequencing the internal transcribed spacer (ITS) rDNA region, partial glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, partial actin (ACT) gene, partial beta-tubulin 2 gene (TUB2), and partial chitin synthase gene (CHS). The obtained sequences have been deposited in GenBank under accession numbers MW581851 and MW581852 (ITS), MW590586 and MW590587 (GAPDH), MW616561 and MW616562 (ACT), MW729380 and MW729381 (TUB2), MW729378 and MW729379 (CHS). The results of Basic Local Alignment Search Tool (BLAST) analysis revealed that the ITS, GAPDH, ACT, TUB2 and CHS sequences of both isolates matched with 100% identity to Colletotrichum fructicola culture collection sequences in GenBank database (JX010165, JX009998, JX009491, JX010405, and JX009866 respectively). These morphological characteristics and molecular analyses allowed the identification of the pathogen as C. fructicola. Koch’s postulates were performed with healthy detached cherry leaves of cultivar namely ‘HongMi’ from Taizhou Academy of Agriculture Sciences. Surface-sterilized leaves were inoculated with five-day-old cultures of C. fructicola mycelial discs of 2 mm in diameter after being wounded with a needle or non-wounded. Control leaves were inoculated with discs of same size PDA agar. Treated leaves were incubated at 25 ℃ in the dark at high relative humidity. Anthracnose symptoms appeared within 3 days both on non-wounded and wounded inoculation approaches. Mock-inoculated controls remained asymptomatic. Biological repetitions were carried out three times. The fungus was reisolated from infected leaves and confirmed as C. fructicola following the methods described above. Until recently, it has been found that C. fructicola can infect tea, apple, pear, Pouteria campechiana in China (Fu et al. 2014; Li et al. 2013; Shi et al. 2018; Yang et al. 2020). To the best of our knowledge, this is the first report of C. fructicola on cherry in China.

Draft genome sequence of Colletotrichum fructicola causing leaf spot on tea plants (Camellia sinensis)

“…In the C. gloeosporioides complex, C. fructicola is one of the most aggressive species, and affects a wide range of plants, such as kiwifruit, cherry, apple and tea plants (Huang, Sheng, et al, 2022;Huang, Wu, et al, 2022;Liu et al, 2022;Lu et al, 2018;Tan et al, 2021;Tang et al, 2021;Zhang et al, 2022). In tea plants, based on the high isolation rate from the main tea region in China, C. fructicola and C. camelliae were identified as the dominant pathogens causing anthracnose disease (Lu et al, 2018).…”

Community Series in Plants and Microbial Communities: Diversity, Pathogens and Biological Control, Volume II

“…doi: 10.1016Biol. 15, /j.cub.2005.047 Batool, W., Shabbir, A., Lin, L., Chen, X., An, Q., He, X., et al (2021). Translation initiation factor eIF4E positively modulates conidiogenesis, appressorium formation, host invasion and stress homeostasis in the filamentous fungi Magnaporthe oryzae.…”

Occurrence and Characterization of Sclerotinia sclerotiorum Causing Fruit Rot on Sweet Cherry in Southern China

“…Sub-optimal cultivation regions located in southern China, such as Hangzhou, have subtropical monsoon climates, for which a rainy spring is one of the climate characteristics. Therefore, the high humidity and suitable temperature in spring lead to high incidences of various fungal diseases in the orchards, such as leaf spot, gummosis, shoot blight, anthracnose, blossom blight, and fruit rot, which seriously restrict the sustainable development of the sweet cherry industry in these regions [ 2 , 14 , 15 , 16 , 17 , 18 , 19 , 20 ].…”