Sources of resistance to Pseudocercospora fijiensis,the cause of black Sigatoka in banana

Black Sigatoka, caused by Pseudocercospora fijiensis, is one of the most devastating diseases of banana. In commercial banana-growing systems, black Sigatoka is primarily managed by fungicides. This mode of disease management is not feasible for resource-limited smallholder farmers. Therefore, bananas resistant to P. fijiensis provide a practical solution for managing the disease, especially under smallholder farming systems. Most banana and plantain hybrids with resistance to P. fijiensis were developed using few sources of resistance, which include Calcutta 4 and Pisang Lilin. To broaden the pool of resistance sources to P. fijiensis, 95 banana accessions were evaluated under field conditions in Sendusu, Uganda. Eleven accessions were resistant to P. fijiensis. Black Sigatoka symptoms did not progress past Stage 2 (narrow brown streaks) in the diploid accessions Pahang (AA), Pisang KRA (AA), Malaccensis 0074 (AA), Long Tavoy (AA), M.A. Truncata (AA), Tani (BB), and Balbisiana (BB), a response similar to the resistant control Calcutta 4. These accessions are potential sources of P. fijiensis resistance and banana breeding programmes can use them to broaden the genetic base for resistance to P. fijiensis.     

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.     

Biochemical responses of coffee resistance against Meloidogyne exigua mediated by silicon

Coffea arabica cultivars Catuaí 44 and IAPAR 59, susceptible and resistant, respectively, to the root knot nematode Meloidogyne exigua, were grown in pots containing Si‐deficient soil amended with either calcium silicate (+Si) or calcium carbonate (?Si). There was an increase of 152 and 100%, respectively, in Si content of root tissue of cvs Catuaí 44 and IAPAR 59 in the +Si compared to the ?Si treatment, but no significant difference between Si treatments for calcium content. Plants, assessed 150 days after inoculation (d.a.i.) showed that the number of galls (NG) and number of eggs (NE) significantly decreased by 16·8 and 28·1% respectively, for susceptible cv. Catuaí 44 in the presence of Si, whilst both NG and NE were significantly lower for cv. IAPAR 59 compared to the susceptible cultivar regardless of Si treatments. In a separate experiment, biochemical assays were carried out 5 and 10 d.a.i. There was no significant difference between Si treatments and cultivars for concentration of total soluble phenolics. The concentration of lignin‐thioglycolic acid (LTGA) derivatives significantly increased by 11·5% in roots of nematode‐inoculated plants of susceptible cv. Catuaí supplied with Si. In roots of inoculated plants of resistant cv. IAPAR 59, the increase was 23 and 10%, respectively, for treatments with and without silicon. Peroxidase (POX), polyphenoloxidase (PPO) and phenylalanine ammonia lyase (PAL) activities significantly increased in roots of inoculated plants compared with roots of non‐inoculated plants, regardless of cultivar or Si treatment. In +Si treatments at 10 d.a.i., POX activity in roots of nematode‐inoculated plants of cvs Catuaí 44 and IAPAR 59 increased by 39·9 and 31·3%, respectively; PPO increased by 54·9 and 56·1%; and PAL activity was also higher at 26·6 and 62·9%. It was concluded that supplying Si to coffee plants increases root resistance against M. exigua by decreasing its reproductive capacity.     

香蕉MaWRKY18的克隆及表达特征分析

为了研究WRKY基因在香蕉与Foc TR4(Fusarium oxysporum f.sp.cubense race 4)互作中作用,利用RACE技术从香蕉中克隆获得了一个WRKY基因全长。序列分析表明,该基因包含完整的开放阅读框,编码275个氨基酸,与香蕉A基因组预测出的(XP_009392153)成员的同源性为100%,因此将其命名为MaWRKY18。MaWRKY18与MaWRKY71(ADR71866)亲缘关系近,聚类在一个分支上。RT-PCR分析表明该基因在香蕉各器官(根、球茎、假茎、叶、花和果实)中均有表达。MaWRKY18受水杨酸和乙烯诱导上调表达,受茉莉酸诱导下调表达。亚细胞定位表明该基因编码的蛋白定位在细胞核中,且C端具有转录激活活性。在Foc TR4接种后,MaWRKY18在感病品种中下调表达,在抗病品种中上调表达。结果表明克隆到的MaWRKY18可能在香蕉与Foc TR4互作过程中介导抗病性。     

Pathogenic variation of Mycosphaerella species infecting banana and plantain in Nigeria

Mycosphaerella species that cause the ‘Sigatoka disease complex’ account for significant yield losses in banana and plantain worldwide. Disease surveys were conducted in the humid forest (HF) and derived savanna (DS) agroecological zones from 2004 to 2006 to determine the distribution of the disease and variation among Mycosphaerella species in Nigeria. Disease prevalence and severity were higher in the HF than in the DS zone, but significant (P < 0·001) differences between agroecological zones were only observed for disease severity. A total of 85 isolates of M. fijiensis and 11 isolates of M. eumusae were collected during the survey and used to characterize the pathogenic structure of Mycosphaerella spp. using a putative host differential cultivar set consisting of Calcutta‐4 (resistant), Valery (intermediate) and Agbagba (highly susceptible). Area under disease progress curve (AUDPC) was higher on all cultivars when inoculated with M. eumusae than with M. fijiensis, but significant (P < 0·05) differences between the two species were only observed on Valery. Based on the rank‐sum method, 8·3% of the isolates were classified as highly aggressive and 46·9% were classified as aggressive. About 11·5% of all the isolates were classified as least aggressive, and all of these were M. fijiensis. The majority of M. eumusae isolates (seven out of 11; 64%) were classified as aggressive. A total of nine pathotype clusters were identified using cluster analysis of AUDPC. At least one M. fijiensis isolate was present in all the nine pathotype clusters, while isolates of M. eumusae were present in six of the nine clusters. Isolates in pathotype clusters III and V were the most aggressive, while those in cluster VIII were the least aggressive. Shannon’s index (H) revealed a more diverse Mycosphaerella collection in the DS zone (H = 1·81) than in the HF (H = 1·50) zone, with M. fijiensis being more diverse than M. eumusae. These results describe the current pathotype structure of Mycosphaerella in Nigeria and provide a useful resource that will facilitate screening of newly developed Musa genotypes for resistance against two important leaf spot diseases of banana and plantain.     

Biochemical responses associated with common bean defence against Sclerotinia sclerotiorum

The aim of this study was to investigate changes in defence compounds of common bean cultivars with different levels of resistance to the fungus Sclerotinia sclerotiorum and determine the relation of the compounds to pathogen tolerance. The lines were inoculated with the pathogen and assessed for enzymatic and non-enzymatic parameters related to plant defence: peroxidases (POX), polyphenol oxidases (PPO), catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX), total soluble phenol and lignin contents. Stem tissue samples were collected from two regions of the plant for biochemical analyses. Stem tissue samples were collected from two regions of the plant for biochemical analyses. In the position one, 5 cm of the stem was collected from the region with necrosis caused by the pathogen, and in the position two, 5 cm of the stem was collected from the end of the position one at the times of 12, 24, 48, 72, 96 and 120 h after inoculation (HAI). Greater lignin and total soluble phenol contents and greater induction of POX and SOD activity in inoculated plants in the region near the inoculation (position one) indicate local activation with later signalling for activation of defence mechanisms in other regions of the plant. The genotype with a greater level of resistance was superior to the susceptible one in regard to lignin production and the activities of POX, APX and SOD defence enzymes. These results suggest that a combination of these defence responses in common bean may contribute to greater plant resistance to the pathogen and that these enzymes have potential use in selection of common bean genotypes.     

Physiological effects of the hydrophilic phytotoxins produced by <Emphasis Type="Italic">Mycosphaerella fijiensis</Emphasis>, the causal agent of black sigatoka in banana plants

Although Mycosphaerella fijiensis, the causal agent of black sigatoka disease of banana, has been known to produce numerous lipophilic host-selective (HSTs) and nonhost selective phytotoxins (non-HSTs), only recently we have reported that the pathogen also produces hydrophilic phytotoxins. Here we examined the effect of light on the toxicity of the hydrophilic phytotoxins and estimated the electrolyte leakage and H₂O₂ and superoxide generation in detached banana leaves to study their mode of action at the cellular level. Nonhost plant species were also tested to determine whether the toxins are HSTs or non-HSTs. Our results suggest that the hydrophilic phytotoxins are non-HSTs, that their phytotoxicity is not light dependent, and that they may act at the plasma membrane by altering permeability through oxidative damage, by inducing ROS production as part of their mechanism of action.     

小偃6号成株期高温抗条锈性遗传分析

为揭示小偃6号抗病机制和培育持久抗病品种,采用常规杂交分析方法,在小麦抽穗期利用小麦条锈菌小种CYR30、CYR32和Su11-4对小偃6号、铭贤169及其杂交F1、F2、F2∶3接种,平均气温达到21℃时对小偃6号进行了抗条锈性调查和遗传分析。结果显示,接种CYR30、CYR32时,F1代表现高感,F2代群体中抗感分离比例符合1 R∶15 S的理论比例。接种Su11-4时,F1代表现高抗,F2代群体中抗感分离比例符合3R∶1S的理论比例。研究表明小偃6号对CYR30、CYR32的抗病性均由2对隐性基因累加作用控制,对Su11-4的抗病性由1对显性基因控制。     

Effect of Juglone on Active Oxygen Species and Antioxidant Enzymes in Susceptible and Partially Resistant Banana Cultivars to Black Leaf Streak Disease

The black leaf streak disease (BLSD), caused by Mycosphaerella fijiensis, is the most destructive disease of bananas and plantains around the world. Breeding for resistance is the most promising strategy to fight this disease especially in small farmer plantations. Mycosphaerella fijiensis produces many phytotoxins such as juglone, which can be used, jointly with field and inoculations under controlled conditions, for screening banana cultivars for BLSD-resistance. This non-host specific phytotoxin has been shown to act on chloroplasts and disturbs the proton electrochemical gradient across the plasmalemma membrane. Moreover, an involvement of the oxidative burst during the interaction has been suggested. The present study was carried out using two cultivars that differed for either their juglone-responses or their resistance to BLSD (cv. Grande Naine susceptible to BLSD and juglone and cv. Fougamou partially resistant to BLSD and highly tolerant to juglone). The production of active oxygen species (AOS) and the enhancement of the enzymatic and/or non-enzymatic AOS-scavenging systems were investigated after treatment of the two cultivars with juglone. The time-course of AOS-production and AOS-scavenging was shown to be the key difference between these two tested cultivars after treatment with juglone. Thus, an early release of AOS (O₂⁻ radical and H₂O₂) and a quick stimulation of a preferment anti-oxidant system (superoxide dismutases, catalases, and peroxidases) was observed for cv. Fougamou as compared to cv. Grande Naine for which a late and weak generation of AOS accompanied by a late stimulation of the anti-oxidant systems were detected.     

Resistance of wheat to Mycosphaerella graminicola involves early and late peaks of gene expression

Large-scale cDNA-AFLP profiling identified numerous genes with increased expression during the resistance response of wheat to the Septoria tritici blotch fungus, Mycosphaerella graminicola. To test whether these genes were associated with resistance responses, primers were designed for the 14 that were most strongly up-regulated, and their levels of expression were measured at 12 time points from 0 to 27 days after inoculation (DAI) in two resistant and two susceptible cultivars of wheat by real-time quantitative polymerase chain reaction. None of these genes was expressed constitutively in the resistant wheat cultivars. Instead, infection of wheat by M. graminicola induced changes in expression of each gene in both resistant and susceptible cultivars over time. The four genes chitinase, phenylalanine ammonia lyase, pathogenesis-related protein PR-1, and peroxidase were induced from about 10- to 60-fold at early stages (3 h–1 DAI) during the incompatible interactions but were not expressed at later time points. Nine other genes (ATPase, brassinosteroid-6-oxidase, peptidylprolyl isomerase, peroxidase 2, 40S ribosomal protein, ADP-glucose pyrophosphorylase, putative protease inhibitor, methionine sulfoxide reductase, and an RNase S-like protein precursor) had bimodal patterns with both early (1–3 DAI) and late (12–24 DAI) peaks of expression in at least one of the resistant cultivars, but low if any induction in the two susceptible cultivars. The remaining gene (a serine carboxypeptidase) had a trimodal pattern of expression in the resistant cultivar Tadinia. These results indicate that the resistance response of wheat to M. graminicola is not completed during the first 24 h after contact with the pathogen, as thought previously, but instead can extend into the period from 18 to 24 DAI when fungal growth increases dramatically in compatible interactions. Many of these genes have a possible function in signal transduction or possibly as regulatory elements. Expression of the PR-1 gene at 12 h after inoculation was much higher in resistant compared to susceptible recombinant-inbred lines (RILs) segregating for the Stb4 and Stb8 genes for resistance. Therefore, analysis of gene expression could provide a faster method for separating resistant from susceptible lines in research programs. Significant differential expression patterns of the defense-related genes between the resistant and susceptible wheat cultivars and RILs after inoculation with M. graminicola suggest that these genes may play a major role in the resistance mechanisms of wheat.     

Powdered neem seed and cake for management of the banana weevil,cosmopolites sordidus, and parasitic nematodes

Field trials were conducted in Kenya with ‘Nakyetengu’, an East African highland banana cultivar (AAA-EA), highly susceptible to banana pests. Regardless of soil fertility levels, incorporation around the plant base of powdered neem(Azadirachta indica A. Juss.) seed or cake at 60-100 g/mat at 4-month intervals, gave better control of the banana weevil,Cosmopolites sordidus (Germar), and of parasitic nematodes, than that achieved with soil application of Furadan 5G (carbofuran) at 60 g/mat at 6-month intervals. Compared with untreated control, fruit yield in most of the neem treatments was significantly higher, particularly during the second cycle of crop production. Neem application conferred a net economic gain, whereas Furadan application proved uneconomical. Application of powdered neem seed or cake at higher rates (200–400 g/mat) at 6-month intervals caused phytotoxicity, resulting in drying up of banana plants before fruiting, or in ‘chokethroat’,i.e., inflorescence emergence failure.     

Phytoanticipins from banana (<Emphasis Type="Italic">Musa acuminata</Emphasis> cv. Grande Naine) plants,with antifungal activity against <Emphasis Type="Italic">Mycosphaerella fijiensis</Emphasis>, the causal agent of black Sigatoka

The previously detected antifungal activity against Mycosphaerella fijiensis of aqueous infusions of healthy banana (Musa acuminata cv. Grande Naine) leaves, suggested the production of phytoprotectants by the plant. The bioassay-guided VLC-purification of the lyophilized infusion of the leaves of 4-month old healthy banana (M. acuminata cv. Grande Naine) plants, resulted in the purification of a phytoanticipin with strong antifungal activity against M. fijiensis Morelet, the causal agent of black Sigatoka, the most destructive and devastating disease of bananas and plantains in the world. The LC-MS analysis of the purified phytoanticipin suggests a steroidal saponin structure with four sugar units attached to the C-3 position of a diosgenin-like aglycone. This represents the first report of phytoanticipins occurring in M. acuminata.     

Biological control mechanisms of D-pinitol against powdery mildew in cucumber

D-pinitol is an effective agent for controlling powdery mildew (Podosphaera xanthii) in cucumber. In this study, we determined the mechanisms of D-pinitol in controlling powdery mildew in cucumber plants. We compared P. xanthii development on cucumber leaf surface treated with D-pinitol or water (2 mg ml⁻¹) at different time points after inoculation. The germinating conidia, hyphae, and conidiophores of the pathogen were severely damaged by D-pinitol at any time of application tested. The highest level of suppression of fungal development was obtained at 3 days after inoculation. The contents of chlorophyll, total phenolics, flavonoid, and gallic acid and its derivatives (GAD); the activities of phenylalanine ammonialyase (PAL), polyphenoloxidase (PPO), peroxidase (POX), and superoxide dismutase (SOD); and the expression of the genes encoding for PR-1, peroxidase (POX), lipoxygenase (LOX1), chitinase (Chit1) were higher in the cucumber leaves treated with D-pinitol and inoculated than in the leaves either treated with D-pinitol or inoculated with the pathogen. These results suggest that D-pinitol triggers several plant defense responses in cucumber leading to pathogen suppression and resistance to powdery mildew.     

Defence responses in flag leaves and spikes of common wheat Triticum aestivum cultivars with contrasting levels of basal resistance to blast caused by Pyricularia oryzae

Wheat blast, caused by Pyricularia oryzae, can cause large yield losses in crops. This study aimed to investigate defence responses in flag leaves and spikes of wheat cultivars BR-18 (moderately resistant) and BRS-Guamirim (susceptible), which differ in their levels of basal resistance. In contrast to cultivar BRS-Guamirim, infected plants of cultivar BR-18 showed more pronounced increases in activities of β-1,3-glucanase and chitinase as well as higher concentrations of lignin-thioglycolic acid derivatives in the flag leaves and total soluble phenolics in the spikes. Polyphenoloxidase activity increased in both flag leaves and spikes in response to fungal infection, regardless of cultivar. Phenylalanine ammonia-lyase (PAL) activity increased in infected flag leaves of both cultivars, especially in BR-18. PAL activity was lower in spikes of infected compared to noninfected plants of both cultivars, although to a lesser extent in BR-18. Compared to BRS-Guamirim, the antioxidative system in both flag leaves and spikes of BR-18 was more efficient in removing reactive oxygen species, reducing cellular damage caused by fungal infection. The lower catalase and peroxidase activities, associated with high superoxide dismutase activity, in flag leaves and spikes of infected BR-18 culminated in a high hydrogen peroxide concentration. The increase in ascorbate peroxidase activity was higher in both flag leaves and spikes of infected plants of BR-18 than in infected BRS-Guamirim. It was concluded that wheat resistance to blast depended on the basal level of resistance of the cultivar, which was mainly associated with the activities of defence enzymes and a more effective antioxidative system.