Characterization and identification of fungicide insensitive Pestalotiopsis-like species pathogenic to tea crop in India

Gray blight, a fungal disease caused by Pestalotiopsis-like species, is a widespread disease affecting tea crop (Camellia sinensis (L.) Kuntze) in many tea-growing countries, including India, resulting in huge losses in tea production. In India, several studies have been conducted to understand the fungal diseases of tea crop, but gray blight has not been well described in major tea growing areas such as in North Bengal, based on its geographic distribution, molecular analysis, or pathogenicity, and even fungicide resistance. The objective of this study was to identify and characterize the causative agents of gray blight disease in symptomatic leaf sample of tea crop collected from 27 tea gardens located in North Bengal, India and to evaluate some common fungicides against them in order to understand the resistance mechanism. In this study, we characterized Pestalotiopsis-like species based on the phylogenies of DNA sequences (internal transcribed spacers) and assessment of conidial characteristics. The study revealed that out of 27 isolates of gray blight pathogens, 17 belonged to the genus Pseudopestalotiopsis (Ps.), six isolates were Neopestalotiopsis, and four were Pestalotiopsis. Two novel species, Ps. thailandica and N. natalensis were introduced through this study. The most frequently isolated genus from C. chinensis was Pseudopestalotiopsis. Pathogenicity tests showed that the isolates displayed significantly different virulence when inoculated onto wounded tea leaves and the mycelial growth rate was positively correlated with pathogenicity (P < 0.01). Based on the 13 ISSR (Inter Simple Sequence Repeat) markers used and principal coordinate analysis, it was found that isolates were very diverse. Out of 27 isolates, IND0P2, DLG0P10, and BHAT0P11 isolates were insensitive against both MBC + M3 (Carbendazim + Mancozeb) and DMI (Hexaconazole) fungicides, while isolates SANY0P18, PAHG0P19, RANG0P24, and SING0P25 were insensitive only against MBC + M3 fungicide. Further, these insensitive isolates were grouped into separate clusters by ISSR, indicating their distinctiveness. However, all the evaluated isolates were susceptible to M1 (copper oxychloride) and another DMI (propiconazole) fungicides. Therefore, to manage gray blight, fungicide resistance management strategies as recommended by Fungicide Resistance Action Committee should be implemented.