First Report of Phytophthora capsici Infection on Bell Peppers (Capsicum annuum L.) from Punjab, Pakistan

Abstract

Bell pepper (Capsicum annuum L.) is one of the extensively cultivated vegetable crop in Punjab, Pakistan. During two years of field surveys, February-November 2016-17, damping off and blight symptoms were observed. Average seedling mortality was recorded as 18.7% while yield loss due to blight was estimated 32 to 41% at mature stages. Maximum blight infection was recorded from the areas frequently flooded with canal irrigation system. At early stages, lesions were noticed on stem portions at soil line level while at crop maturity stages blight symptoms were noted. Leaves were blanched and wilted while fruits were covered with white mold. Masses of sporangia were evident on and inside the infected fruits under humid conditions. A total of twelve isolates were recovered from infected root, stem and fruit portions on rye agar media (Caten and Jinks, 1968) incubated at 25^oC under fluorescent light. Papillated sporangia were averaged 42 ± 2.6 X 27 ± 1.7 μm in size (range 27 - 52 × 23 - 36 μm). Oospores were produced on 20% V8 agar and were spherical 22 ± 1.4 μm in diameter (range 14 to 27 μm) while average pedicels length was recorded as 58 ± 12.5 μm (range 13 to 120 μm). These observations were similar to those described for P. capsici (Cocoa, 1988). DNA was extracted using Cetyl Trimethylammonium Bromide (CTAB) method and the internal transcribed spacer regions (ITS1 and ITS2) were amplified by polymerase chain reaction (White et al., 1990). The amplicons were purified and sequenced in both directions (GenBank Accession No. MF322868 and MF322869). BLAST analysis revealed these isolates showed 99% identity with ITS sequences of Phytophthora capsici (KM369964 and KU518782). Pathogenicity assay was performed on healthy bell pepper seedlings with five repeats. Soil was flooded with 20ml sporangial suspension (1 x 10³ sporangia/ml) in pots containing seedlings while 5ml suspension was sprayed until run off on mature plants (Hyder et al., 2018). A set of uninoculated seedlings was used as control. Pots were kept in dew chamber for 10-20 days at 25±2 ^oC. Seedling mortality was observed five days after inoculation while at later stage plants develop brown-to-black stem lesions with white mycelial growth on leaves. These symptoms were identical to the P. capsici infections in field. Consistent re-isolations of P. capsici confirm its association with the disease. To our knowledge, this is the first report of Phytophthora blight on bell pepper from Pakistan