Biocontrol of a root rot of kale by <Emphasis Type="Italic">Muscodor albus</Emphasis> strain MFC2

Pythium ultimum is an oomycetous root rot pathogen that causes significant crop production losses on many crops including kale (Brassica oleracea), an economically important vegetable in Thailand. An endophytic fungus from Thailand designated Muscodor albus MFC2 controlled P. ultimum both in vitro and on kale seedlings grown under outdoor conditions via the production of volatile antibiotics. Ten-day old M. albus MFC2 PDA cultures killed P. ultimum in vitro. Thoroughly mixing three PDA plates of 10-day old M. albus MFC2 into a 500 g mixture of commercial soil and field soil did not adversely affect kale seed germination. The same amount of M. albus MFC2 could restore seedling emergence in P. ultimum inoculated soil to a level close to that of a non-infested control. In addition, M. albus MFC2 did not cause any disease symptoms, but rather seemed to promote the growth of kale in the presence or absence of P. ultimum for up to eight weeks after planting.     

Pseudomonas cichorii as the causal agent of midrib rot,an emerging disease of greenhouse-grown butterhead lettuce in Flanders

Bacterial midrib rot of greenhouse-grown butterhead lettuce (Lactuca sativa L. var. capitata) is an emerging disease in Flanders (Belgium) and fluorescent pseudomonads are suspected to play an important role in the disease. Isolations from infected lettuces, collected from 14 commercial greenhouses in Flanders, yielded 149 isolates that were characterized polyphasically, which included morphological characteristics, pigmentation, pathogenicity tests by both injection and spraying of lettuce, LOPAT characteristics, FAME analysis, BOX-PCR fingerprinting, 16S rRNA and rpoB gene sequencing, as well as DNA–DNA hybridization. Ninety-eight isolates (66%) exhibited a fluorescent pigmentation and were associated with the genus Pseudomonas. Fifty-five of them induced an HR⁺ (hypersensitive reaction in tobacco leaves) response. The other 43 fluorescent isolates were most probably saprophytic bacteria and about half of them were able to cause rot on potato tuber slices. BOX-PCR genomic fingerprinting was used to assess the genetic diversity of the Pseudomonas midrib rot isolates. The delineated BOX-PCR patterns matched quite well with Pseudomonas morphotypes defined on the basis of colony appearance and variation in fluorescent pigmentation. 16S rRNA and rpoB gene sequence analyses allowed most of the fluorescent isolates to be allocated to Pseudomonas, and they belonged to either the Pseudomonas fluorescens group, Pseudomonas putida group, or the Pseudomonas cichorii/syringae group. In particular, the isolates allocated to this latter group constituted the vast majority of HR⁺ isolates and were identified as P. cichorii by DNA–DNA hybridization. They were demonstrated by spray-inoculation tests on greenhouse-grown lettuce to induce the midrib rot disease and could be re-isolated from lesions of inoculated plants. Four HR⁺ non-fluorescent isolates associated with one sample that showed an atypical midrib rot were identified as Dickeya sp.     

Suppression of Ripe Rot on ‘Zesy002’ Kiwifruit with Commercial Agrochemicals

Ripe rot caused by Botryosphaeria dothidea is one of the serious diseases of postharvest kiwifruit. In order to control ripe rot on Actinidia chinensis cultivar ‘Zesy002’, several commercial agrofungicides were selected by an antifungal test on an artificial medium. Furthermore, disease suppression by the selected fungicides was evaluated on the kiwifruit by inoculation with a conidial suspension of B. dothidea. On the artificial media containing boscalid + fludioxonil was shown to be the most effective antifungal activity. However, in the bio-test pyraclostrobin + boscalid and iminoctadine-tris were the most effective agrochemicals on the fruit. On the other hand, the infection structures of B. dothidea on kiwifruit treated with pyraclostrobin + boscalid were observed with a fluorescent microscope. Most of the fungal conidia had not germinated on the kiwifruit treated with the agrochemicals whereas on the untreated fruit the fungal conidia had mostly germinated. Electron microscopy of the fine structures showed morphological changes to the conidia and branch of hyphae on the kiwifruit pre-treated with pyraclostrobin + boscalid, indicating its suppression effect on fungal growth. Based on this observation, it is suggested that ripe rot by B. dothidea may be suppressed through the inhibition of conidial germination on the kiwifruit treated with the agrochemicals.     

Genetic Diversity of Populations of Monilinia fructicola (Fungi,Ascomycota, Helotiales) from China

ABSTRACT. The genetic variation among 128 isolates of Monilinia fructicola (Fungi, Ascomycota, Helotiales) from China was analyzed using Inter‐Simple Sequence Repeat (ISSR) markers and compared with those of samples from California, USA and New Zealand. A total of 72 reproducible DNA fragments were scored, of which 87.5% (63/72) were polymorphic. The Nei's gene diversity and Shannon's diversity indices of three Chinese regional populations were very similar to that from California. However, several differences were observed among geographic populations of M. fructicola from both within China and between China and California. The analysis of molecular variance (AMOVA) of isolates from different geographic locations suggested that most of the observed genetic variation was found within populations. Results of this study are inconsistent with the hypothesis that the Chinese populations of M. fructicola were derived from a single or few recent migrants from other countries. Instead, our results suggest that M. fructicola has been in China long before its first official recording in 2003.     

Fusarium culmorum is a single phylogenetic species based on multilocus sequence analysis

Fusarium culmorum is a major pathogen of wheat and barley causing head blight and crown rot in cooler temperate climates of Australia, Europe, West Asia and North Africa. To better understand its evolutionary history we partially sequenced single copy nuclear genes encoding translation elongation factor 1-α (TEF), reductase (RED) and phosphate permease (PHO) in 100 F. culmorum isolates with 11 isolates of Fusarium crookwellense, Fusarium graminearum and Fusarium pseudograminearum. Phylogenetic analysis of multilocus sequence (MLS) data using Bayesian inference and maximum parsimony analysis showed that F. culmorum from wheat is a single phylogenetic species with no significant linkage disequilibrium and little or no lineage development along geographic origin. Both MLS and TEF and RED gene sequence analysis separated the four Fusarium species used and delineated three to four groups within the F. culmorum clade. But the PHO gene could not completely resolve isolates into their respective species. Fixation index and gene flow suggest significant genetic exchange between the isolates from distant geographic regions. A lack of strong lineage structure despite the geographic separation of the three collections indicates a frequently recombining species and/or widespread distribution of genotypes due to international trade, tourism and long-range dispersal of macroconidia. Moreover, the two mating type genes were present in equal proportion among the F. culmorum collection used in this study, leaving open the possibility of sexual reproduction.     

The use of biological selenium nanoparticles to suppress Triticum aestivum L. crown and root rot diseases induced by Fusarium species and improve yield under drought and heat stress

Fusarium species threaten wheat crops around the world and cause global losses. The global trend is toward using biological materials such as selenium (Se) in nano form to control these fungi. Bulk selenium is toxic and harmful at high doses; however, selenium nanoparticles are safe; therefore, the aim of this study to employ the biological selenium nanoparticles (BioSeNPs) synthesized by Lactobacillus acidophilus ML14 in controlling wheat crown and root rot diseases (CRDs) induced by Fusarium spp., especially Fusarium culmorum and Fusarium graminearum, and their reflection on the growth and productivity of wheat. The ability of BioSeNPs to suppress the development and propagation of F. culmorum and F. graminearum and the CRDs incidence were also investigated. The obtained BioSeNPs were spherical with a size of 46 nm and a net charge of –23.48. The BioSeNPs significantly scavenged 88 and 92% of DPP? and ABT? radicals and successfully inhibited the fungal growth in the range of 20–40 µg/mL; these biological activities were related to the small size of BioSeNPs and the phenolic content in their suspension. Under greenhouse conditions, the wheat supplemented with BioSeNPs (100 µg/mL) was significantly reduced the incidence of CRDs by 75% and considerably enhanced plant growth, grain quantity and quality by 5–40%. Also, photosynthetic pigments and gas exchange parameters were significantly increased as compared to chemical selenium nanoparticles (Che-SeNPs) and control. This study results could be recommended the use of BioSeNPs (100 µg/mL) in reducing CRDs incidence and severity in wheat plants, enhancing their tolerance with drought and heat stress, and increasing their growth and productivity as compared to control and Che-SeNPs.     

Development of SCAR markers linked to three disease resistances based on AFLP within Nicotiana tabacum L

Amplified fragment length polymorphism (AFLP) was conducted on a set of 92 Nicotiana tabacum L. accessions from diverse types (flue-cured, dark air-cured, burley, oriental, and cigar wrapper) and breeding origins to identify markers associated with disease resistances. Eleven primer combinations were required to identify 33 polymorphic fragments. This allowed the identification of 92% of these accessions, and yielded sufficient information for building a neighbor joining tree. Clusters of accessions with common traits or breeding origins were observed. An important part of this polymorphism could be related to interspecific introgressions from other Nicotiana species, performed during the breeding history of N. tabacum to confer resistance to pathogens. Seven fragments were associated with three different resistances: two for the blue-mold (Peronospora tabacina Adam) resistance derived from Nicotiana debneyi Domin, two for the Va gene (Potato Virus Y susceptibility), and three for the black root rot (Chalara elegans) resistance of N. debneyi origin. Some of these markers were converted into sequence characterized amplified region markers, and validated on recombinant inbred lines or doubled-haploid lines.     

Interactions of soil pH,nutrients and moisture on phytophthora root rot of avocado

Summary This experiment employed a factorial design combining 4 soil pH levels, 3 soil moisture levels, with and without the addition ofPhytophthora cinnamomi to the soil to evaluate the conditions that lead to Phytophthora root rot of avocado.An inverse relation between soil pH and leaf production (and root-weight) was observed in nondiseased plants. In soil infested withP. cinnamomi, plant growth and root weights were much depressed by low soil pH, and especially by low soil pH coupled with high soil moisture contents. These interactions were statistically highly significant. Root weights in pots withP. cinnamomi were closely related to the incidence of disease. A disease index was used to visually assess the conditions of roots. Isolation of the pathogen from diseased plant roots confirmed the accuracy of the disease index.A process of elimination suggsts that favorable soil Ca level and not high pHper se was responsible for disease suppression and that the devastating effects of low soil pH was produced by high Mn (and possibly Al) and associated low levels of Ca and P in soil solutions, which led to breakdown of biological control mechanisms.Journal Series No. 2801, Hawaii Institute of Tropical Agriculture and Human Resources.