Effect of Sampling Time, Quantification Method, and Tree Sample Pooling on Root Quantities in South African Avocado Orchards.

is a destructive soilborne pathogen causing Phytophthora root rot on avocados worldwide. Little is known about the effect of root sampling time, root quantification method (quantitative real-time PCR [qPCR] versus baiting), and tree sample pooling strategies on the quantification of the pathogen in roots in avocado orchard trees. This was investigated in six avocado orchards in two climatically different production regions (Mooketsi and Letaba) in the Limpopo Province, South Africa, over a 2-year period. Two different tree sample pooling strategies, consisting of either a four-pooled group (four groups each containing five pooled trees) or a single-pooled group (20 trees pooled) per 1 ha, were both shown to be suitable for quantifying in tree roots using qPCR or root baiting. root quantities from the two tree sample pooling strategies were significantly correlated for both quantification methods. Both quantification methods were suitable for quantifying the pathogen in roots, although qPCR was superior to root baiting at identifying significant differences in quantities among root sampling time points. The effect of sampling time was dependent on the investigated year. In 2017, root quantities, which were only evaluated using qPCR, did not reveal a consistent trend of a specific sampling time yielding the highest root quantities for most of the orchards. However, five of the orchards in 2018, based on the qPCR analyses, contained significantly higher root quantities in May (late autumn) than in March (early autumn), August (late winter), and October/November (late spring). In 2018, root DNA quantities were significantly positively correlated with the number of soil temperature hours at 20 to 24 and 20 to 29°C 2 months preceding the root sampling dates and negatively correlated with the number of hours at 15 to 19°C 2 months preceding root sampling. Our study has identified root quantification methods and tree sample pooling strategies, which will be useful for understanding the biology of the pathogen and when disease management strategies should be in place.