Limited Evidence for Accumulation of Latent Infections of Canker-Causing Pathogens in Shoots of Stone Fruit and Nut Crops in California.

Prevalence of latent infections of the canker-causing fungi Botryosphaeria dothidea and species of Cytospora, Diplodia, Lasiodiplodia, Neofusicoccum, and Phomopsis in young shoots of almond, prune, and walnut trees in California was studied to test the hypotheses that latent infections accumulate from current-season shoots to 1-year-old shoots in the orchard and there are distinct associations among pathogen taxa present as latent infections in the same shoot. Samples of newly emerged and 1-year-old shoots were periodically collected in each almond, prune, and walnut orchard for two growing seasons. A real-time quantitative PCR assay was used to quantify latent infection with three parameters: incidence, molecular severity, and latent infection index. Diplodia spp. were absent from most samples. For almond, Lasiodiplodia spp. and Cytospora spp. were detected with a maximum incidence >90%, while B. dothidea and Neofusicoccum spp. incidence was <20% in most cases. In prune orchards, the incidence levels of B. dothidea were >50% in most cases, while those of Cytospora spp. and Lasiodiplodia spp. were 30 to 60% and 30 to 100%, respectively. For walnut, many samplings showed higher incidence in 1-year-old (30 to 80%) than in newly emerged shoots (10 to 50%). Accumulation of latent infection between the two shoot age classes was detected in only a few cases. The percentages of samples showing coexistence of two, three, and four pathogen taxa in the same shoot were 20 to 25, <10, and <5%, respectively. Pairwise associations among pathogen taxa in the same shoot were significant in many cases.

Interaction Between Diaporthe rhusicola and Neofusicoccum mediterraneum Causing Branch Dieback and Fruit Blight of English Walnut in California, and the Effect of Pruning Wounds on the Infection.

Botryosphaeriaceae and Diaporthaceae species are the causal agents of branch dieback of English walnut in California. In this study, the effects of the interaction between Neofusicoccum mediterraneum and Diaporthe rhusicola were evaluated in vitro by using mycelial plugs or spore suspensions and in vivo by inoculating shoots and epicarps (hulls) of walnut. Single inoculations of each species and different coinfection treatments were performed under laboratory or field conditions. The influence of shoot age and susceptibility of bark or pith tissues to N. mediterraneum and D. rhusicola infection after pruning was also evaluated. In in vitro experiments, spore germination of D. rhusicola was significantly (P < 0.0001) reduced in the presence of N. mediterraneum spores. When D. rhusicola was inoculated at 4 days before N. mediterraneum, a delay in lesion development in shoots and hulls was observed compared with the other two interaction treatments. One- to 2-year-old shoots were more susceptible to infection and colonization by N. mediterraneum than 3- to 4-year-old shoots. In young shoots, inoculation in the pith tissue resulted in longer lesions than those observed on shoots inoculated in the bark. No significant differences were observed between the development of internal or external necrosis and the age of the shoots, or the susceptibility of bark and pith to D. rhusicola infection. This information is essential to better understanding the complex situation of this walnut disease toward developing control management strategies.

Evaluation of Two Disease Warning Systems for Botryosphaeria Panicle and Shoot Blight of California Pistachio and Efficient Control Based on Early-Season Sprays.

Two empirical models to predict infection events were evaluated for control of Botryosphaeria panicle and shoot blight, caused by a Fusicoccum sp., as well as the effectiveness of early-season fungicide sprays on the control of this disease of pistachio. A model incorporating wetness duration was superior to one based solely on duration of rains ≥1 mm/h for ≥4 h and with temperature ≥11°C. The wetness duration threshold (W) for rain events ≥4 mm at a given temperature (T) for high-risk infection events was W = -7.8 + 397/T and the threshold for medium-risk events was W = -6.9 + 220/T. Wet periods interrupted by ≤12 h were added together to calculate W. In two orchards with high levels of inoculum, one high-risk event resulted in 20 to 23% blighted fruit at harvest and two or three high-risk events resulted in 31 to 80% blighted fruit. Latent infections were 0 to 1% in instances where only low-risk events (one to two events) occurred prior to collection of pistachio fruit for determination of latent infections and were 17 to 36% with one to three high-risk events. Early-season fungicide sprays in April to May effectively controlled panicle and shoot blight when applied up to 12 days before predicted infection events or 5 days after.

Antagonism of Paenibacillus lentimorbus to Botryosphaeria dothidea and Biological Control of Panicle and Shoot Blight of Pistachio.

A potential microbial fungicide, Paenibacillus lentimorbus isolate CBCA-2, against Botryosphaeria dothidea, the pistachio panicle and shoot blight fungus, was obtained from healthy pistachio leaves by both in vitro and in vivo screening techniques. CBCA-2 caused 100% inhibition of pycnidiospore germination after 24 h incubation at 25°C. Malformation of pycnidiospores and hyphae, and lysis and swollen pycnidiospores of B. dothidea occurred in the presence of cell suspensions of CBCA-2. Among the five media tested, nutrient yeast dextrose broth significantly increased the production of antifungal compounds. Application of culture filtrates of CBCA-2 suppressed disease on detached pistachio leaves, but washed bacterial cells did not inhibit lesion development. Development of lesions on excised dormant stems was inhibited only when the culture filtrate was applied before fungal inoculation. Survival of the CBCA-2 after treatment with azoxystrobin (Abound), benomyl (Benlate), tebuconazole (Elite), propiconazole (Break), or trifloxystrobin (Flint) at the highest recommended concentration was not affected, but survival was affected by iprodione (Rovral). Spraying a suspension of CBCA-2 on pruning wounds before inoculation with pycnidiospores of B. dothidea significantly reduced infection compared with the unsprayed, inoculated controls.