First report of leaf spot disease on oil palm caused by Pestalotiopsis theae in Thailand

A leaf spot on oil palm, caused by Pestalotiopsis theae, was found in a plantation of Elaeis guineensis for the first time in the world in Chiang Mai Province, Thailand. The fungus was isolated from lesions on leaves, and its pathogenicity was confirmed. Pathogenicity tests showed that P. theae could infect E. guineensis, which developed the same symptoms after inoculation as those observed naturally in the field. The fungus was identified based on morphological characteristics and confirmed using comparisons of DNA sequences of internal transcribed spacer (ITS)1, ITS2 and 5.8S rDNA. This report is the first on oil palm leaf spot disease caused by P. theae.     

Effect of tricyclazole on growth and secondary metabolism in Pyricularia oryzae

Tricyclazole [5-methyl-1,2,4-triazolo (3,4-b)-benzothiazole] controls rice blast disease caused by Pyricularia oryzae at concentrations (5–10 μg/ml) which do not inhibit growth of the pathogen in vitro. However, concentrations of 1 μg/ml or less inhibit melanin formation in the fungus. Production of pyriculol by the pathogen is usually enhanced by 10 μg/ml of tricyclazole, whereas production of 3,4-dihydro-4,8-dihydroxy-1(2H)-naphthalenone is strongly inhibited or markedly reduced and delayed. Evidence suggests that tricyclazole blocks aspects of polyketide metabolism in P. oryzae which may have a role in pathogenicity.     

Involvement of OsNPR1/NH1 in rice basal resistance to blast fungus Magnaporthe oryzae

Rice blast disease, caused by the fungus Magnaporthe oryzae, is a major threat to worldwide rice production. Plant basal resistance is activated by virulent pathogens in susceptible host plants. OsNPR1/NH1, a rice homolog of NPR1 that is the key regulator of systemic acquired resistance in Arabidopsis thaliana, was shown to be involved in the resistance of rice to bacterial blight disease caused by Xanthomonas oryzae pv. oryzae and benzothiadiazole (BTH)-induced blast resistance. However, the role of OsNPR1/NH1 in rice basal resistance to blast fungus M. oryzae remains uncertain. In this study, the OsNPR1 gene was isolated and identified from rice cultivar Gui99. Transgenic Gui99 rice plants harbouring OsNPR1-RNAi were generated, and the OsNPR1-RNAi plants were significantly more susceptible to M. oryzae infection. Northern hybridization analysis showed that the expression of pathogenesis-related (PR) genes, such as PR-1a, PBZ1, CHI, GLU, and PAL, was significantly suppressed in the OsNPR1-RNAi plants. Consistently, overexpression of OsNPR1 in rice cultivars Gui99 and TP309 conferred significantly enhanced resistance to M. oryzae and increased expression of the above-mentioned PR genes. These results revealed that OsNPR1 is involved in rice basal resistance to the blast pathogen M. oryzae, thus providing new insights into the role of OsNPR1 in rice disease resistance.     

Infection by Magnaporthe oryzae chrysovirus 1 strain A triggers reduced virulence and pathogenic race conversion of its host fungus, <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

Magnaporthe oryzae chrysovirus 1 strain A (MoCV1-A) is associated with an impaired growth phenotype of its host fungus, Magnaporthe oryzae. In this report, we assayed the virulence and pathogenicity of MoCV1-A-infected and MoCV1-A-free M. oryzae on rice plants. MoCV1-A infection did not affect virulence-associated fungal traits, such as conidial germination and appressorium formation. However, after punch inoculation of leaves on rice plants, MoCV1-A-infected strain formed smaller lesions than the MoCV1-A-free strain did on all rice varieties tested, showing that MoCV1-A infection resulted in reduced virulence of host fungi in rice plants. In contrast, after spray inoculation of rice seedlings, in some cases, MoCV1-A-infected and MoCV1-A-free strains caused different lesion types (resistance to susceptible, or vice versa) on individual international differential rice varieties. However, we did not find any gain/loss of the fungal avirulence genes by PCR, suggesting that MoCV1-A infection can convert the pathogenicity of the host M. oryzae from avirulence to virulence, or from virulence to avirulence, depending on the rice variety. We also confirmed the correlation of these race conversion events and invasive hyphae growth of the fungi in a leaf sheath inoculation assay. These data suggested that MoCV1-A infection generally confers hypovirulence to the fungal host and could be a driving force to generate physiological diversity, including pathogenic races.     

The occurrence of postharvest gray-mold rot of sweet persimmon caused by <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in Korea

In this study, we identified the causative agent of postharvest gray-mold rot in sweet persimmon fruit that were collected from Gangneung, Gangwon Province, Korea in October 2016. Symptoms included extensive growth of mycelia on post harvested fruit. The fungus was isolated from infected fruit and cultured on potato dextrose agar (PDA). For identification of the fungus, we examined morphology characteristics and rDNA sequencing analysis of the fungus and confirmed its pathogenicity according to Koch’s postulates. The results of morphological examinations, pathogenicity tests, 5.8S rDNA sequences of the internal transcribed spacer regions (ITS1 and ITS4) and the five nuclear protein-coding genes G3PDH, HSP60, RPB2, MS547 and TUB revealed that the causal agent of postharvest gray-mold rot on sweet persimmon fruit in Korea was Botrytis cinerea.     

黑龙江省水稻种质抗瘟性及稻瘟病菌致病性分析

为明确黑龙江省水稻种质抗性及稻瘟病菌的致病性,以黑龙江省8个水稻品种、24个单基因系作为供试材料,120株稻瘟病菌株作为接种体,采用喷雾接种法测定了各供试水稻的抗瘟性及稻瘟病菌的致病性。结果表明,水稻品种对2010年和2011年菌株的抗性频率分别在31.67%~68.33%和21.67%~55.00%之间,2010年最好的抗性品种为松粳12,2011年最好的抗性品种为五优稻4和东农425;松粳12东农425组合联合抗病性最好。水稻单基因系对2010年和2011年菌株的抗性频率分别在10.00%~90.00%和5.00%~86.67%之间,抗性最好的单基因系分别为IRBLzt-T(Pi-zt)和IRBLz5-CA(Pi-z5);松粳12、东农425和龙粳22的基因聚合效果最好。2010年和2011年菌株对抗瘟基因群的致病率分别在8.33%~95.83%和25.00%~95.83%之间;无毒基因总出现频率分别为461和412次。研究表明,水稻种质抗性受菌株致病性影响较大,但高抗种质相对稳定,基因聚合方式更适宜当地品种抗性改良。     

水直播条件下黑龙江省不同稻区稻瘟病菌致病性分析

为明确水直播条件下黑龙江省不同稻区稻瘟病菌Magnaporthe oryzae的致病性分化情况,以24个抗瘟单基因系品种为寄主,来源于2017—2018年黑龙江省水直播稻田的242株稻瘟病菌菌株为接种体,采用离体划伤方法接种,记录病斑反应型,计算有效致病菌株率和抗性频率,并进行聚类分析。结果显示,在水直播条件下,2017年,黑龙江省南部和中东部稻区稻瘟病菌菌株对抗瘟单基因系品种的有效致病菌株率介于8.33%~95.83%和20.83%~95.83%之间,无毒基因出现频率分别为575次和622次;2018年,南部和中东部稻区稻瘟病菌菌株对鉴别体系的有效致病菌株率介于29.17%~95.83%和20.83%~91.67%之间,无毒基因出现频率分别为536次和571次。2017年,黑龙江省南部和中东部稻区稻瘟病菌菌株的致病性相似系数介于0.15~1.00和0.14~1.00之间,以致病性相似系数0.40为阈值,可将菌株分别划分为5个类群和6个类群;2018年,南部和中东部稻区菌株的致病性相似系数介于0.15~0.93和0.26~1.00之间,以致病性相似系数0.40为阈值,可将菌株分别划分为5个类群和4个类群。2017年,抗瘟单基因系品种对黑龙江省南部和中东部稻区稻瘟病菌菌株的抗性频率介于11.29%~88.71%和10.77%~86.15%之间,其中抗瘟单基因系品种IRBL9-W（Pi-9）和IRBLz5-CA（Pi-z5）抗性表现最好;2018年,抗瘟单基因系品种对南部和中东部稻区菌株的抗性频率介于10.34%~82.67%和15.79%~85.96%之间,其中抗瘟单基因系品种IRBL9-W（Pi-9）和IRBLz5-CA（Pi-z5）抗性表现最好。表明水直播条件下黑龙江省稻瘟病菌致病性分化剧烈,稻瘟病菌整体致病力较强,但仍有部分水稻种质抗性较好且相对稳定,基因聚合后抗性会得到进一步提升。     

Mechanisms of date palm resistance to Bayoud disease: Current state of knowledge and research prospects

The Bayoud disease, caused by Fusarium oxysporum f. sp. albedinis (Foa), represents a major limiting factor of date palm culture in Morocco and constitutes a serious threat to the date palm plantations in Algeria and all other countries. Efficient disease prevention requires the development of resistant cultivars. In Morocco, among the cultivars listed, only six appear to be resistant to Bayoud disease, but they produce poor quality fruit. Thus, the Moroccan program of date palm genetic improvement is based on directed crossing between resistant cultivars and susceptible cultivars with good date quality traits to select resistant genotypes producing high quality fruits. In addition to the separation of the resistance to Bayoud disease and quality of the fruits characters, this breeding program is really complex due to the sex separation in the date palm, the duration of juvenile phase which is very long, and the lifespan of the date palm which requires a durable polygenic resistance. Then, the selected genotypes must be of female sex, of good date quality, and possess effective defense mechanisms against the pathogen. Moreover, the selection of the date palm resistance must necessarily take into account the mechanisms of pathogen aggressiveness. In this review, we will present and discuss studies developed on the Bayoud disease of the date palm, particularly on the disease control, the biochemical and molecular markers of resistance, the program of date palm genetic improvement of resistance, the Foa pathogenicity factors, and host defense mechanisms. It will also highlight the recent studies that showed that differential behaviour of the resistant and susceptible cultivars was not related to a difference of induction of the defense mechanisms, but to the suppression of their elicitation in the susceptible cultivars.     

Characterization and pathogenicity of <Emphasis Type="Italic">Bipolaris peregianensis</Emphasis>: the causal organism for leaf spot of hybrid bermudagrass in China

Several golf courses established with hybrid bermudagrass (Cynodon dactylon × C. transvaalensis) were surveyed from April 2011 through April 2015 in Hainan Province, China. The hybrid bermudagrass in these golf courses showed a new leaf spot disease, and a filamentous fungus was consistently recovered from the infected leaves. Based on the morphological characteristics of colony color and appearance, shapes of conidiophores and conidia as well as sequences of internal transcribed spacer regions (ITS) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), the fungus was identified as Bipolaris peregianensis. The pathogenicity test conducted on healthy hybrid bermudagrass produced leaf spot symptoms one week post inoculation. B. peregianensis mycelia grew in a temperature range of 5 to 35 °C with the optimum temperature being 28 °C.     

Pathogenicity,host range and activities of a secondary metabolite and enzyme from Myrothecium roridum on water hyacinth from Thailand

Leaf blight disease of water hyacinth was observed and collected from different geographical areas of Thailand. The disease is caused by a fungal pathogen that was identified as Myrothecium roridum by using its morphological characteristics. The most effective fungal strains were evaluated for pathogenicity on water hyacinth under greenhouse and natural conditions. Myrothecium roridum isolate, Kamphaeng Sean Campus (KKFC) 448, was found to be the most virulent. Different fungal formulations were evaluated for their level of control of water hyacinth. The results showed that spore suspensions with 10% palm oil or 1% Tween 20 caused a higher level of disease severity, compared to spores applied in water alone. The host range of KKFC 448 was evaluated by using 77 plant species that belong to 40 plant families. The fungus did not cause disease on 74 economically important plants but did produce disease signs on water hyacinth and two other aquatic weeds, duckweed and water lettuce. Leaf blight occurs on water hyacinth leaves after being treated with crude extracts of M. roridum and it was indicated that secondary metabolites were released from the fungal mycelia. Myrothecium roridum that was grown on boiled paddy rice produced β‐1,4‐exoglucanase, β‐1,4‐endoglucanase, β‐glucosidase, xylanase and pectinase more than Mroridum that was grown on potato dextrose agar. The results indicated that M. roridum is a pathogen of water hyacinth and the fungus is capable of producing different enzymatic activities on potato dextrose agar and boiled paddy rice, which might be important for infection.     

A boll rot of cotton caused by Corynespora Cassiicola in Tamil Nadu,India

A new boll rot disease of cotton (Gossypium hirsutum L.) cv. MCU 9 was observed in Tamil Nadu, India, in 1988. The causal organism was isolated in pure culture and identified asCorynespora cassiicola (Berk. & Curt.) Wei. The isolate, which on cotton is pathogenic only to bolls, is considered to be different from a strain ofC. Cassiicola known elsewhere causing leaf spot of cotton. The pathogenicity of this fungus on several other important crops was also tested. The fungus produced leaf spot and stem rot diseases on, respectively, eight and six host plants. Root rot, after inoculation, was observed only inVigna sinensis, Arachis hypogaea andSesamum indicium.     

Antifungal activity of a novel compound purified from the culture filtrate of <Emphasis Type="Italic">Biscogniauxia</Emphasis> sp. O821 against the rice blast fungus <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

Rice blast is a devastating fungal disease resulting in major losses to rice crops. Owing to continuous acquisition of resistance by the causal fungus, several fungicide chemicals are no longer effective. Therefore, there is a need to identify natural components and develop new agents to control fungal pathogens. We previously demonstrated that the culture filtrate of Biscogniauxia sp. O821 inhibited infection behavior of Magnaporthe oryzae and subsequent blast lesion formation. In the present study, we isolated a new compound, (3aS,4aR,8aS,9aR)-3a-hydroxy-8a-methyl-3,5-dimethylenedecahydronaphto[2,3-b]furan-2(3H)-one (HDFO), from the culture filtrate of Biscogniauxia sp. O821 and determined its molecular weight as 248. The HDFO structure was determined by electrospray ionization-mass spectrometry and nuclear magnetic resonance spectroscopy after purification with column chromatography and high-performance liquid chromatography. The structure of this antifungal compound was similar to that of alantolactone and isoalantolactone. The growth inhibition zone against M. oryzae in presence of HDFO was observed at Rf 0.5–0.6 on a thin layer chromatography plate. HDFO inhibited conidial germination of M. oryzae in a dose-dependent manner (1–200 ppm). Furthermore, blast lesion formation was significantly suppressed by HDFO at over 5 ppm. These results suggest that HDFO from the culture filtrate of Biscogniauxia sp. O821 can protect rice from rice blast disease caused by M. oryzae. This is the first report that HDFO produced by Biscogniauxia sp. can serve as an antifungal compound against M. oryzae.     

Site‐directed mutagenesis of the cytochrome b gene and development of diagnostic methods for identifying QoI resistance of rice blast fungus

BACKGROUND: It is possible that a single nucleotide polymorphism (SNP) (G143A mutation) in the cytochrome b gene could confer resistance to quinone outside inhibiting (QoI) fungicides (strobilurins) in rice blast fungus because this mutation caused a high level of resistance to fungicides such as azoxystrobin in Pyricularia grisea Sacc. and other fungal plant pathogens. The aim of this study was to survey Magnaporthe oryzae B Couch sp. nov. isolates in Japan for resistance to QoIs, and to try to develop molecular detection methods for QoI resistance. RESULTS: A survey on the QoI resistance among M. oryzae isolates from rice was conducted in Japan. A total of 813 single‐spore isolates of M. oryzae were tested for their sensitivity to azoxystrobin using a mycelial growth test on PDA. QoI fungicide resistance was not found among these isolates. The introduction of G143A mutation into a plasmid containing the cytochrome b gene sequence of rice blast fungus was achieved by site‐directed mutagenesis. Molecular diagnostic methods were developed for identifying QoI resistance in rice blast fungus using the plasmid construct. CONCLUSION: As the management of rice blast disease is often dependent on chemicals, the rational design of control programmes requires a proper understanding of the fungicide resistance phenomenon in field populations of the pathogen. Mutation of the cytochrome b gene of rice blast fungus would be specifically detected from diseased leaves and seeds using the molecular methods developed in this study. Copyright © 2009 Society of Chemical Industry     

Effects of panicle development stage and temperature on rice panicle blast infection by Magnaporthe oryzae and visualization of its infection process

Panicle blast, caused by the fungus Magnaporthe oryzae (syn. Pyricularia oryzae), directly contributes to yield loss in the field. The effects of panicle development stage and temperature on panicle blast were studied and the infection process of M. oryzae in panicles was visualized. Rice panicles at different development stages from three rice cultivars were inoculated with a conidial suspension in vitro. The rice cultivar Lijiangxintuanheigu was highly susceptible to panicle blast at 5 days postinoculation (dpi) when the pulvinus distance was 15–20 cm. Nanjing 9108 was moderately susceptible to panicle blast when the pulvinus distance was 8–10 cm, but Yliangyou 800 was resistant. The effect of temperature on panicle blast was determined under 22–35 °C temperature treatments. Inoculated panicles placed at temperatures of 28 and 30 °C showed the highest lesion grade based on lesion length at 5 dpi. The infection process of M. oryzae in rice panicles was observed by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). M. oryzae initially formed the appressorium to invade through the epidermis of rice panicles at 24 hours postinoculation (hpi). As the disease progressed, the invasive hyphae formed dense mycelial networks in the inner parenchyma cells at 60 hpi. Our results will contribute to the understanding of panicle development stage and temperature effects on panicle blast and improve resistance evaluation methods. Additionally, visualization of the infection process by CLSM and TEM are valuable methods to observe M. oryzae invasive hyphae inside rice panicle cells.     

Antifungal activity of <Emphasis Type="Italic">Biscogniauxia</Emphasis> sp. culture filtrates against the rice pathogen <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

An ethyl acetate extract of a culture filtrate (ECF) from an unidentified fungal isolate O821 was evaluated for antifungal activity against the rice pathogen Magnaporthe oryzae. The O821-ECF significantly inhibited spore germination, appressorium formation, and mycelial growth of M. oryzae, and its antifungal activity was heat-stable. It also significantly suppressed the number and size of blast lesions. In an analysis of the ITS sequence of this isolate, it shared similarities with species of the fungus Biscogniauxia. These results suggest that isolate O821 of the genus Biscogniauxia produces a heat-stable antifungal compound(s) in its culture filtrate.     

Genetic and molecular analyses of the incompatibility between Lolium isolates of Pyricularia oryzae and wheat

Pyricularia oryzae isolates from Lolium spp. (annual ryegrass and perennial ryegrass) show evidence of recent events in evolution of this fungus. A wheat blast isolate found in Kentucky in 2011 was assumed to originate from annual ryegrass isolates. Genetic analyses revealed that the incompatibility between a Lolium isolate and common wheat cultivars is controlled by two gene pairs, Rmg6–A1 with a strong effect and R2-A2 with a weak effect, implying that this incompatibility is conditioned by simple gene-for-gene interactions. Disruption of the A1 avirulence gene led the Lolium isolate to gain virulence on common wheat. These results suggest a mechanism for host jumping by the blast fungus.     

三份重要水稻资源的稻瘟病抗性鉴定及不同评价指标间的相关性分析

为明确杂交水稻育种骨干亲本93-11、纹枯病重要抗源YSBR1及水稻重要基础研究的遗传转化材料Kitaake共3份重要水稻资源的稻瘟病抗谱及不同评价指标间的相关性,通过喷雾接种法,采用国际水稻稻瘟病抗性评价分级标准、致病率、病斑数量和病斑长度4个评价指标,分析了这3份重要水稻资源对36个稻瘟病菌生理小种的抗性。结果表明这3份水稻资源材料的病情指数主要集中于4和5级,但针对不同的生理小种其致病率存在较大差异。同时,93-11、YSBR1和Kitaake产生的病斑平均长度分别为0.56、0.54和0.66 cm,病斑平均数量分别为6.50、4.10和4.80个。Spearman相关性分析证明所采用的4个稻瘟病抗性评价指标之间均呈极显著正相关。表明93-11、YSBR1和Kitaake均对多数稻瘟病生理小种表现出抗性,但抗谱各不相同,且采用的4个评价指标间无显著差异。     

Fusarium solani f. sp. cucurbitae race 1, a potential pathogen of grafted watermelon production in Spain*

A crown, root and fruit rot of squash (Cucurbita maxima and Cucurbita moschata) has been observed in eastern provinces of Spain over the past 4 years. Isolations from the crown of symptomatic plants and fruits yielded primarily a Fusarium solani that was identified as F. solani f. sp. cucurbitae race 1 on the basis of pathogenicity tests and disease symptoms in the field. In Spain, more than 90% of watermelon plants are grafted, using different Cucurbita hybrids (C. maxima × C. moschata) as rootstocks. In 1998, some grafted watermelon plants were first found to be affected by F. s. cucurbitae race 1. The purpose of this study was to evaluate the pathogenicity of this fungus on several rootstocks commonly used for grafting watermelon (Brava, Titan, Shintoza, RS‐841, TZ‐148 and TW‐1) in order to prevent a possible spread of this fungus that could cause serious economic losses in watermelon production. None of them proved to be resistant.     

Functional analysis of the G-protein α subunits FGA1 and FGA3 in the banana pathogen Fusarium oxysporum f. sp. cubense

The asexual fungus Fusarium oxysporum f. sp. cubense (Foc) is the causal agent of fusarium wilt in bananas (Musa spp.). This fungus poses a threat to banana production throughout the world. Here, two Foc genes, fga1 and fga3, were functionally characterized. These genes encode proteins homologous to the G-protein α subunits GPA1 from Saccharomyces cerevisiae and MAGC from Magnaporthe grisea, respectively. The deletion of fga1 leads to a phenotypic defect in colony morphology and reductions in vegetative growth, conidiation and pathogenicity against the banana plant (Musa spp. cv. Brazil), which was not observed for the Δfga3 deletion mutant. Intriguingly, both Δfga1 and Δfga3 deletion mutants showed declines in intracellular cyclic AMP levels and increases in heat resistance, suggesting that FGA1 regulates growth, development, pathogenicity, and heat resistance, whereas FGA3 modulates heat resistance, potentially through the cAMP-dependent protein kinase A pathway. These findings offer insights into the roles of the G-protein α subunits in the development and pathogenicity of the fungus Foc.