Pre-plant bacterisation: a strategy for delivery of beneficial endophytic bacteria and production of disease-free plantlets of black pepper (Pipernigrum L.)

Naturally occurring endophytic bacteria from black pepper vines were found to exhibit strong antagonistic activities against Phytophthora capsici and Radopholus similis. In order to deliver these bacterial strains, as well as to produce disease-free plantlets of black pepper, a pre-plant stem and root bacterisation was standardised. Stem bacterisation with endophytic Pseudomonas spp. was found to suppress P. capsici infection (over 90% reduction in lesion length) on cut shoots. Pre-plant root bacterisation with Pseudomonas aeruginosa, Pseudomonas putida and Bacillus megaterium yielded over 60% of plantlets free from P. capsici infection on roots. Curtobacterium luteum and B. megaterium recorded over 70% reduction of nematode population in soil with concomitant production of over 65% of nematode-free plantlets. Besides protecting the plants from the pathogens, the bacteria were also found to enhance the growth of rooted cuttings. The biocontrol potential of the above endophytic bacteria and their exploitation for disease management in the black pepper nursery are discussed.     

Evaluation of fatty acid methyl ester profile and amplified fragment length polymorphism analyses to distinguish five species of Phytophthora associated with ornamental plants

Fatty acid methyl ester (FAME) profiles and amplified fragment length polymorphisms (AFLPs) were evaluated as tools for identifying species of Phytophthora. Five isolates of each of Phytophthora cactorum, Phytophthora citrophthora, Phytophthora cinnamomi, Phytophthora nicotianae and Phytophthora cryptogea were subjected to both analyses to examine variation among and within species. In FAME analysis, isolates of P. cactorum, P. cinnamomi and P.nicotianae were clustered by species, but isolates of P. citrophthora and P.cryptogea were divided into multiple clusters based on greater variations within these two species. The AFLP analysis differentiated all five species of Phytophthora. The five isolates of each species were grouped in a separate terminal cluster, but diversity within a species cluster varied considerably with variation greater in P. cryptogea and P. citrophthora. Comparing the dendrograms based on FAME and AFLP analyses, the overall patterns of both were similar. The P. cactorum cluster was distinct from clusters of the other four species, which formed one large cluster. The higher values of percentages of polymorphic loci and gene diversity in AFLP analysis substantiated diversity observed among isolates of P. citrophthora and P. cryptogea in FAME and AFLP dendrograms. Both FAME and AFLP appear to be useful tools for identifying species of Phytophthora, but only AFLP analysis has potential to study genetic and phylogenetic relationships within and among species in this genus.     

First Report of Phytophthora Foliar Blight on Florida Hopbush (Dodonaea viscosa) in Italy

During 2010, a new foliar blight was detected on potted Dodonaea viscosa cv. Purpurea plants in two nurseries in Catania (Italy). On the basis of morphological and cultural features, the pathogen was identified as Phytophthora palmivora. The internal transcribed spacer (ITS)‐rDNA sequence of a representative Phytophthora isolate from hopbush showed 99% identity with other ITS sequences of different P. palmivora isolates available in GenBank, thus confirming the morpho‐cultural identification. Koch’s postulates were fulfilled by pathogenicity tests on potted D. viscosa cv. Purpurea seedlings. To our knowledge, this is the first report of P. palmivora foliar blight disease on D. viscosa.     

Conditions for Infection of Apple by Phytophthora syringae

The processes leading to Phytophthora fruit rot were divided into two main stages for the purposes of investigating the effects of temperature and duration of wet periods on pathogen development: oospore germination and infection of fruit by zoospores. It was found that the first stage was markedly affected by temperature over the range 10–20°C and required a wet period of 4–7 days. At 18 and 20°C, activation was low regardless of the length of the wet period. Once oospore germination (first stage) had occurred, free water was necessary for only a few hours for fruit infection (second stage) to occur, but the incidence of infection rose rapidly over the first 48 h, regardless of temperature over the range 10–20°C. From the data obtained, mathematical models were produced relating the incidence of Phytophthora fruit rot to the two weather variables. These models can be used to develop a weather‐based risk assessment system for the disease.     

Resistance to crown rot (Phytophthora cactorum) in strawberry cultivars and in offspring from crosses between cultivars differing in susceptibility to the disease

During a period of five years (1998‐2002), 26 strawberry cultivars and five selections were tested for resistance to crown rot. Cold stored plants inoculated with zoospores of Phytophthora cactorum were used in all experiments. The results showed that resistance to P. cactorum varies greatly between cultivars, and the most resistant ones were Senga Sengana, Induka, Melody, Glima and Bogota, while the most susceptible were Tamella, Inga, Evita and Jonsok. The results were compared to those obtained by other authors, and there were clear indications that the genetic background affects the degree of susceptibility to P. cactorum. Many of the most resistant cultivars descend from Senga Sengana (e.g. Bounty, Glima, Induka and Melody), and several of the most susceptible cultivars have common parentage (e.g. Inga, Tamella, Elsanta and Evita). In a resistance test of the progeny from two separate crosses between a resistant and a susceptible cultivar 61% and 65% of the offspring were intermediately to very susceptible, and 17% and 13% were resistant at the level of Senga Sengana. Hence, there is a high risk of losing offspring with high levels of resistance if selection for resistance to crown rot is not performed at an early stage in a breeding programme.     

Role of the cyclic lipopeptide massetolide A in biological control of Phytophthora infestans and in colonization of tomato plants by Pseudomonas fluorescens

Pseudomonas strains have shown promising results in biological control of late blight caused by Phytophthora infestans. However, the mechanism(s) and metabolites involved are in many cases poorly understood. Here, the role of the cyclic lipopeptide massetolide A of Pseudomonas fluorescens SS101 in biocontrol of tomato late blight was examined. Pseudomonas fluorescens SS101 was effective in preventing infection of tomato (Lycopersicon esculentum) leaves by P. infestans and significantly reduced the expansion of existing late blight lesions. Massetolide A was an important component of the activity of P. fluorescens SS101, since the massA-mutant was significantly less effective in biocontrol, and purified massetolide A provided significant control of P. infestans, both locally and systemically via induced resistance. Assays with nahG transgenic plants indicated that the systemic resistance response induced by SS101 or massetolide A was independent of salicylic acid signalling. Strain SS101 colonized the roots of tomato seedlings significantly better than its massA-mutant, indicating that massetolide A was an important trait in plant colonization. This study shows that the cyclic lipopeptide surfactant massetolide A is a metabolite with versatile functions in the ecology of P. fluorescens SS101 and in interactions with tomato plants and the late blight pathogen P. infestans.