Mode of resistance to respiration inhibitors at the cytochrome bc1 enzyme complex of Mycosphaerella fijiensis field isolates

Field isolates of Mycosphaerella fijiensis, causing black Sigatoka of banana, were characterised for their sensitivity to different inhibitors of the cytochrome bc₁ enzyme complex (Qo respiration inhibitors, strobilurin fungicides), using physiological, biochemical and molecular genetic methods. Strobilurin‐resistant isolates exhibited very high resistance factors both in mycelial growth inhibition and NADH consumption assays. Cross‐resistance was observed among all Qo inhibitors, including trifloxystrobin, azoxystrobin, famoxadone, strobilurin B and myxothiazol. However, the Qi and the cytochrome aa₃ inhibitors, antimycin A and potassium cyanide, respectively, were not cross‐resistant to Qo inhibitors. In sensitive but not in resistant isolates, mixtures of Qo inhibitors and SHAM, an inhibitor of the alternative oxidase (AOX), were more active than the components alone, indicating that the alternative pathway is essential in metabolism, but not causal for resistance. In the cell‐free NADH‐consumption assay, the Qo inhibitors affected the sensitive but not the resistant isolates, suggesting that AOX was not active in sub‐mitochondrial particles. In whole cells, however, the AOX has a basic expression level and is probably not inducible by trifloxystrobin. Sequencing of the cytochrome b gene of sensitive and resistant M fijiensis isolates revealed a difference in the nucleotide sequence leading to a single amino acid change from glycine to alanine at position 143 in the resistant isolate. This change is known to occur also in the naturally tolerant basidiomycete Mycena galopoda. It is suggested that the field isolates of M fijiensis can acquire resistance to Qo inhibitors due to a target site alteration with a single base pair change. Resistant isolates do not seem to contain a mixture of mutated and non‐mutated DNA, indicating a complete selection of resistant mitochondria and a maternally donated mode of resistance. © 2000 Society of Chemical Industry     

Genetic structure of Mycosphaerella fijiensis populations from Australia, Papua New Guinea and the Pacific Islands

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of Mycosphaerella fijiensis , the cause of black leaf streak (black Sigatoka) disease of banana and plantain, in the Torres Strait, Papua New Guinea (PNG), and the Pacific Islands. A moderate level of genetic variation was observed in all populations with genotypic diversity values of 60–78% of the theoretical maximum, and gene diversity ( H ) values between 0·269 and 0·336. All populations were at gametic equilibrium, and with the high level of genotypic diversity observed this indicated that sexual reproduction has a major role in the genetic structure of the M. fijiensis populations examined. Population differentiation was tested on several hierarchical scales. No evidence of population differentiation was observed between sites on Mer Island. A moderate level of population differentiation was observed within the Torres Strait, between Badu and Mer Islands ( F _ST = 0·097). On a regional scale, the greatest differentiation was found between the populations of the Torres Strait and the Pacific. Populations from these regions were more closely related to the PNG population than to each other, suggesting they were founded in separate events from the same population.     

Genetic variability of Pseudocercospora fijiensis,the black Sigatoka pathogen of banana (Musa spp.) in Mexico

Pseudocercospora (previously known as Mycosphaerella) fijiensis causes black Sigatoka disease in banana (Musa spp.) and is considered to be the most devastating pathogen of this crop worldwide. To improve knowledge of its evolutionary patterns, this study determined the genetic variability of populations from two regions of Mexico: Central Pacific (Colima and Michoacan) and Southern (Chiapas, Tabasco and Oaxaca), using 10 simple sequence repeat (SSR) loci and the MAT-specific PCR assay. Both mating types were present in all regions under study, with frequencies of 63% MAT1-1 and 37% MAT1-2. The SSR markers showed an average of three alleles per locus, resulting in 34 alleles in total. The genetic diversity (H_T) was 0.3308, but at the local level (H_S) ranged from 0.0976 (Colima) to 0.2228 (Oaxaca). However, the genotypic diversity was usually high (H′ > 2.4, S > 0.89). Cluster analysis grouped the isolates into five clusters with high statistical support (au > 80%), suggesting a geographic organization of the genetic variability of P. fijiensis; AMOVA, the minimum spanning tree and the population structure analysis supported this result, and all data indicated that the major genetic differences were between the different populations under analysis. Thus, the high level of genetic variability in P. fijiensis is attributed partly to a high rate of sexual reproduction, and also to a strong evolutionary capacity coupled with isolation due to limited genetic flow between distant populations. Both possibilities could be playing a relevant role in population differentiation of the pathogen.     

Tackling black leaf streak disease and soil fertility constraints to enable the expansion of plantain production to grassland in the humid tropics

In Central Africa, plantain is traditionally grown after a forest fallow. Given increasing urban demand and a lack of forest fallows near urban centres, as well as poor roads and environmental concerns to reduce pressure on forests, research is needed to identify suitable shade, fertility management and cultivars to shift production of plantain to grasslands and to reduce losses to diseases such as black leaf streak disease (BLSD). Effects of light level (full, 67%, 33% light), and nitrogen (N)-amendment on BLSD-tolerant (FHIA-21) and BLSD-susceptible (Batard) cultivars planted on soil from paired grassland and forest sites were determined. BLSD and growth were monitored until 5 months after planting. Three months after planting, leaf area attacked on cultivar FHIA-21 was less than half that on Batard. Plants grown under 33% and 67% light had less leaf area attacked (2.9% and 4.6%, respectively) than those grown in full light (7.3%). Leaf area and dry matter (DM) were higher under shade and when grown on forest soils. Compared to growing BLSD-susceptible plantain on forested land under shade, a shift onto grasslands and a reduction in shade use is predicted to reduce yields. Using cultivar FHIA-21 may limit, but not eliminate, yield loss.     

Relationship between the Spray Droplet Density of Two Protectant Fungicides and the Germination of Mycosphaerella fijiensis Ascospores on Banana Leaf Surfaces

The effect of fungicide spray droplet density (droplet cm^-2), droplet size, and proximity of the spray droplet deposit to fungal spores was investigated with Mycosphaerella fijiensis ascospores on the banana (Musa AAA) leaf surface for two contact fungicides: chlorothalonil and mancozeb. When droplet size was maintained at a volume median diameter (VMD) of 250 μm while total spray volume per hectare changed, M. fijiensis ascospore germination on the leaf surface fell below 1% for both fungicides at a droplet deposit density of 30 droplet cm^-2. At a droplet deposit density of 50 droplet cm^-2, no ascospores germinated in either fungicide treatment. When both droplet size and droplet cm^-2 varied while spray volume was fixed at 20 litre ha^-1, ascospore germination reached 0% at 10 droplet cm^-2 (VMD=602 μm) for both fungicides. At lower droplet densities (2–5 droplet cm^-2 VMD=989 μm and 804 μm respectively), ascospore germination on the mancozeb-treated leaves was significantly lower than on the chlorothalonil-treated leaves. The zone of inhibition surrounding a fungicide droplet deposit (VMD=250 μm) on the leaf surface was estimated to extend 1·02 mm beyond the visible edge of the spray droplet deposit for chlorothalonil and 1·29 mm for mancozeb. The efficacy of fungicide spray droplet deposit densities which are lower than currently recommended for low-volume, aerial applications of protectant fungicides was confirmed in an analysis of leaf samples recovered after commercial applications in a banana plantation. Calibrating agricultural spray aircraft to deliver fungicide spray droplets with a mean droplet deposit density of 30 droplet cm^-2 and a VMD between 300 and 400 μm will probably reduce spray drift, increase deposition efficiency on crop foliage, and enhance disease control compared to aircraft calibrated to spray finer droplets. © 1997 SCI.     

Distribution of Pseudocercospora species causing Sigatoka leaf diseases of banana in Uganda and Tanzania

Sigatoka leaf diseases are a major constraint to banana production. A survey was conducted in Tanzania and Uganda to assess the distribution of Pseudocercospora species and severity of Sigatoka leaf diseases. Pseudocercospora species were identified using species-specific primers. Sigatoka-like leaf diseases were observed in all farms and on all cultivars, but disease severity varied significantly (P < 0.001) between countries, districts/regions within countries, altitudinal ranges and banana cultivars. In all regions except Kilimanjaro, P. fijiensis, the causal agent of black Sigatoka, was the only pathogen associated with Sigatoka disease. Mycosphaerella musae was associated with Sigatoka-like symptoms in Kilimanjaro region. Black Sigatoka disease was more severe in Uganda, with a mean disease severity index (DSI) of 37.5%, than in Tanzania (DSI = 19.9%). In Uganda, black Sigatoka disease was equally severe in Luwero district (mean DSI = 40.4%) and Mbarara district (mean DSI = 37.9%). In Tanzania, black Sigatoka was most severe in Kagera region (mean DSI = 29.2%) and least in Mbeya region (mean DSI = 11.5%). Pseudocercospora fijiensis, the most devastating sigatoka pathogen, was detected at altitudes of up to 1877 m a.s.l. This range expansion of P. fijiensis, previously confined to altitudes lower than 1350 m a.s.l. in East Africa, is of concern, especially for smallholder banana farmers growing the susceptible East African Highland bananas (EAHB). Among the banana varieties sampled, the EAHB, FHIA hybrids and Mchare were the most susceptible. Here, the loss of resistance in Yangambi KM5, a banana variety previously resistant to P. fijiensis, is reported for the first time.     

香蕉褐缘灰斑病菌检测技术研究进展

香蕉褐缘灰斑病严重影响香蕉产量和品质,是香蕉生产上的重要生物灾害之一.香蕉褐缘灰斑病的早期诊断、检疫及防治的重要基础与前提是建立快速、灵敏的检测技术,从而准确及时鉴定出病原菌种类.本文介绍了香蕉褐缘灰斑病菌的种类和分类,概述了国内外香蕉褐缘灰斑病菌检测方法的研究进展,总结和归纳了传统、血清学、DNA指纹图谱、常规PCR...     

三种猎物对小黑瓢虫成虫繁殖及寿命的影响

在实验室条件下,以自然猎物烟粉虱Bemisiatabaci为对照,研究了替代猎物红蜘蛛Tetranychusnr.fi jiensis和桃蚜Myzuspersicae(Sulzer)对小黑瓢虫成虫繁殖及寿命的影响,结果表明:(1)小黑瓢虫以蚜虫为猎物时,成虫不产卵,但可延长寿命;以红蜘蛛为猎物时,成虫可产卵。与自然猎物烟粉虱相比,小黑瓢虫取食替代猎物红蜘蛛的产卵前期延长,总产卵量及平均每雌每天产卵量减少,而两者之间的性比、产卵率及产卵期无显著差异。(2)三种猎物条件下雌虫寿命之间显著性无差异,而雄虫寿命之间的差异达显著水平,t测定法进一步表明,雄虫寿命在蚜虫与红蜘蛛之间差异显著,而在蚜虫与烟粉虱之间、以及烟粉虱与红蜘蛛之间均无显著差异。