黑龙江省部分地区稻瘟病菌致病性分析及鉴别体系优化

为明确哈尔滨市及鸡西市稻瘟病菌致病性变化情况并优化鉴别体系,以24个水稻单基因系为鉴别体系,对2006—2008年采自2市的稻瘟病菌菌株进行致病型划分,并应用聚类分析方法完成鉴别体系优化。结果表明,应用水稻单基因系鉴别体系可将哈尔滨市2006—2008年采集的稻瘟病菌菌株划分为34、12和27个致病型,致病性相似系数分别为0.32~1.00、0~1.00和0.20~1.00;水稻单基因系对其抗性频率分别为2.86%~97.14%、8.33%~100.00%和10.00%~96.67%;鸡西市2006—2008年采集的菌株可划分为26、19和20个致病型,致病性相似系数分别为0.47~0.92、0.15~0.86和0.39~1.00;水稻单基因系对其抗性频率分别为3.85%~96.15%、5.26%~73.68%和4.76%~95.24%。适用于哈尔滨市的优化鉴别体系包括Pi-9、Pi-11、Pi-a、Pi-ks、Pi-5、Pi-i、Pi-sh、Pi-3、Pi-km和Pi-ta共10个基因,2006—2008年累计方差贡献率分别为86.15%、98.77%和87.39%;适用于鸡西市的优化鉴别体系包括Pi-9、Pi-11、Pi-a、Pi-ks、Pi-5、Pi-i、Pi-sh、Pi-1、Pi-7和Pi-t共10个基因,2006—2008年累计方差贡献率分别为82.58%、94.55%和90.37%。应用优化后的鉴别体系可将哈尔滨市2006—2008年采集的菌株划分为22、12和23个致病型,将鸡西市2006—2008年采集的菌株划分为22、14和18个致病型。     

Recovery and characterization of asexual recombinants ofMagnaporthe grisea

Eleven nitrate non-utilizing (nit) mutants were recovered from six field isolates ofMagnaporthe grisea that had different degrees of sensitivity to the blasticide kitazin P (iprobenfos, IBP). Allnit mutants were resistant to chlorate and there were no significant differences in hyphal growth rate, conidial production, sensitivity to IBP, and pathogenicity betweennit mutants and their parent isolates.nit phenotypes and IBP-resistance were used as two independent genetic markers to study asexual recombination inM. grisea. Asexual recombinants were recovered from the heterokaryotic mycelium of three inter-strain pairings, namely, DY2-3(nitl-LR) + A7-3 (nitM-S); A7-3 (nitM-S) + F4-2 (nitl-MR); and F4-2 (nitl-MR) + DY2-4 (nitA-LR), at a frequency of 7.31%, 14.00% and 8.63%, respectively. The growth rate, conidial production and pathogenicity of asexual recombinants were similar to those of parental strains. We concluded that asexual recombination resulting from hyphal fusion might contribute to variability inM. grisea. http://www.phytoparasitica.org posting Nov. 14, 2005.     

Effect of glyphosate on the sprouting of Cyperus rotundus L. tubers

Post-emergence applications of glyphosate [N-(phosphonomethyl)glycine] have been shown not to eradicate purple nutsedge (Cyperus rotundus L.) in the field. It was not known if this was due to failure to control emerged plants or if dormant tubers produced new plants after application. Studies with individual plants were conducted in screenhouse facilities to determine the effects of glyphosate rate, time for translocation, area of foliage treated, and shade on the sprouting ability of tubers attached to treated plants. Rates of 1.5–2.0 kg/ha glyphosate inhibited tuber sprouting; 72 h were required for complete translocation at 1.0 kg/ha whereas 36 h were sufficient at 2.0 kg/ha. Treating less than all of the foliage reduced foliar control and increased tuber sprouting. Shading treated plants reduced control of the foliage but did not affect glyphosate translocation to the tubers. These studies showed that glyphosate kills C. rotundus foliage and the tubers attached to treated plants. Therefore, regrowth after glyphosate application under field conditions is due to dormant tubers which sprout after treatment.     

Effect of glyphosate on tryptamine production and Sekiguchi lesion formation in rice infected with Magnaporthe grisea

When exposed to light, the Sekiguchi lesion (sl) rice mutant has an enhanced resistance to Magnaporthe grisea infection responsible for Sekiguchi lesion formation and tryptamine accumulation. Glyphosate [N-(phosphonomethyl) glycine] pretreatment suppressed Sekiguchi lesion formation and tryptamine accumulation in the sl mutant after M. grisea infection even under light. This inhibition by glyphosate was blocked by the supply of exogenous tryptophan, but not by exogenous phenylalanine. In glyphosate-pretreated leaves, 5-enol-pyruvyl-shikimate-3-phosphate synthase gene expression and tryptophan biosynthesis were significantly suppressed. During tryptophan starvation, catalase activity was maintained at a high level even under light, leading to the suppression of H₂O₂ generation and DNA fragmentation. These results show a strong relationship between the tryptophan and tryptamine pathways in the induction of light-enhanced resistance to M. grisea infection in the sl mutant.     

DNA fingerprinting of Pyricularia grisea by rep-PCR using a single primer based on the terminal inverted repeat from either of the transposable elements Pot2 and MGR586

We investigated the use of single primers complementary to sequences in the terminal inverted repeat (TIR) of either Pot2 or MGR586, transposable elements found in Pyricularia grisea, for DNA fingerprinting by repetitive-element-based polymerase chain reaction (rep-PCR). Under standard amplification conditions, rep-PCR with each single primer generated distinct fingerprint patterns among rice-infecting P. grisea isolates collected in Japan. With the Pot2-TIR primer, bands ranging in size from 0.2 to 8 kb and in number from 8 to 13 per isolate were amplified. Although fewer bands were amplified with the MGR586-TIR primer, this molecular technique should be more reliable to identify and classify P. grisea isolates by combining the data of fingerprint patterns from each TIR primer. In a cluster analysis based on DNA fingerprints from this rep-PCR with the Pot2-TIR primer, 10 reference isolates and 12 field isolates from Saga Prefecture in 2002 were separated into six clonal lineages. We also demonstrated that the 12 field isolates belonged to one clonal lineage. Thus, this rep-PCR method using the single primer Pot2-TIR will be useful for the analysis of the population structure of rice blast pathogens.     

Effect of inhibitors of melanin biosynthesis on appressorial penetration and reductive reactions in Pyricularia oryzae and Pyricularia grisea

Appressorial penetration of onion epidermal walls by wild-type strains P-2 and O-42 of Pyricularia oryzae was more sensitive to fthalide (4,5,6,7-tetrachlorophthalide) than penetration by wildtype strain 4091-5-8 and black mutant strain BL-3 of Pyricularia grisea. Cerulenin completely blocked appressorial penetration by P. oryzae strain P-2 or P. grisea strain BL-3, but penetration capacity of these appressoria could be largely restored with 0.1 mM scytalone. Fthalide and tricyclazole inhibited the conversion of 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN) to 1,3,8-trihydroxynaphthalene (1,3,8-THN) and 1,8-dihydroxynaphthalene (1,8-DHN) as well as the conversion of scytalone to 1,8-DHN by cell-free extracts of P. oryzae or P. grisea. These inhibitors blocked the NADPH-dependent reductase reactions involved in the conversion of 1,3,6,8-THN to scytalone and the conversion of 1,3,8-THN to vermelone in the melanin biosynthetic pathway to 1,8-DHN. The two reductase reactions in extracts from P. oryzae P-2 were about 10 times as sensitive to fthalide or tricyclazole as those in extracts of P. grisea. Reductase activity with either 1,3,6,8-THN or 1,3,8-THN as substrates was present in only trace amounts in cell-free extracts of the buff mutant, P. oryzae P-2 m-1.     

Possible roles and functions of LPL1 gene encoding lysophospholipase during early infection by Magnaporthe grisea

The rice blast fungus Magnaporthe grisea differentiates appressoria, which are required to attack its rice plant host. Clone A26, tentatively named LPL1, was previously found to be homologous to the known lysophospholipase genes from our subtractive cDNA library. The LPL1 protein had a consensus motif (GxSxG) and a catalytic triad (S, D, H) of esterases in the deduced amino acid sequence, and the protein expressed in Escherichia coli had lysophospholipase activity. To clarify the functions and possible roles of LPL1, the gene was disrupted by targeted gene replacement. The ΔLPL1 mutants formed fewer appressoria on the hydrophobic surface of GelBond film, and the appressoria had reduced turgor pressure and penetration into cells of the leaf sheath. The ΔLPL1 mutants and wild-type differentiated normal appressoria on other artificial substrata such as polycarbonate plate and on rice leaf sheath. Cytological analysis of the appressoria indicated that ΔLPL1 mutants had a delay in the disappearance of lipid droplets. These findings imply that LPL1, phospholipid metabolism, or both are involved in glycerol biosynthesis and accumulation to generate turgor pressure in the appressorium. LPL1 was, however, dispensable for full pathogenicity, suggesting that other complementary pathways or similar genes related to phospholipid metabolism probably function in M. grisea.     

Site factors and management influence short-term host resistance to spruce budworm, Choristoneura fumiferana (Clem.), in a species-specific manner

BACKGROUND: This study examined the interactions between thinning and soil drainage classes on the resistance of balsam fir, Abies balsamea (L.) Mill, white spruce, Picea glauca (Moench) Voss, and black spruce, P. mariana (Mill.) BSP, to spruce budworm, Choristoneura fumiferana (Clem.), defoliation 1 year after treatment. To estimate host tree resistance, foliage production and larval foliage consumption were determined to generate an index of resistance quantifying the amount of residual foliage available for photosynthesis after insect defoliation. RESULTS: Significant interactions on tree resistance and foliage chemistry were detected between thinning and soil drainage in balsam fir. Drainage class affected spruce budworm performance, foliar chemistry and balsam fir resistance to spruce budworm, whereas no effect was found in white spruce. Thinning had a significant effect on the foliar chemistry of balsam fir and white spruce, but no effect on black spruce. CONCLUSION: Thinning reduced balsam fir resistance to spruce budworm defoliation. This response is due to increased defoliation linked to reduction in concentrations of certain monoterpenes, and a decrease in foliage production, except on hydric drainage, demonstrating the importance of drainage class to tree resistance. The results suggest that the use of Bacillus thuringiensis might be required when conducting stand thinning during spruce budworm outbreaks. Copyright © 2011 Society of Chemical Industry     

广东省稻瘟病菌生理小种的消长动态

１９９０～１９９７年,用中国稻瘟病菌鉴别寄主鉴定了广东省稻瘟病菌单孢菌株１５７４份,共鉴定出８群４３个生理小种。其中ＺＣ群一直是广东省的优势种群,其出现频率平均为５０.２％;ＺＣ_１３一直是优势小种,其出现频率平均为２７.４％,其次是ＺＧ_１（１８.８％）、ＺＣ_１５（１４.５％）和ＺＢ_１３（９.１％）小种。不同地区不同年份优势小种的类型及其组成不同。     

Oxidative stress induces high transcription of Rhm52 and Rhm54, novel genes identified as being involved in recombinational repair in Magnaporthe grisea

The first two recombinational repair genes of Magnaporthe grisea were cloned. Analysis of the deduced amino acid sequences revealed that Rhm52 and Rhm54 are Saccharomyces cerevisiae RAD52 and RAD54 homologs, respectively. Phenotypic complementation testing of these genes showed their function in recombinational repair. Both genes were in single copies in M. grisea genome. Expression of these genes was induced by methyl methanesulfonate and ultraviolet radiation as known for other homologs of the RAD52 epistasis group. Higher induction of both genes by oxidative stress and heat shock indicated the probability for recombinational repair during the infection cycle of M. grisea. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB091332 and AB091330     

Expression profiling of rice genes in early defense responses to blast and bacterial blight pathogens using cDNA microarray

In order to understand the defense machinery in the model cereal crop rice, we performed a large-scale analysis of rice gene expression in response to rice blast Magnaporthe grisea (M. grisea) or Magnaporthe oryzae and bacterial blight Xanthomonas oryzae pv. oryzae (Xoo) during the early incompatible and compatible interactions. Using a gene chip containing 10 254 rice cDNAs representing 9240 unique genes, we identified 794 and 612 genes differentially expressed in the incompatible and compatible rice–M. grisea interactions, respectively, with 274 genes co-regulated during both interactions. In the rice–Xoo pathosystem, 454 and 498 differentially expressed genes were identified in the incompatible and compatible interactions, respectively, including 237 co-regulated genes in the both interactions. By clustering differentially regulated genes from all these interactions, we identified 29 co-regulated genes in the all four interactions, and 86 and 74 co-regulated genes in the two incompatible and two compatible interactions, respectively. These differentially expressed genes could be classified into three categories, including M. grisea- and Xoo-regulated, M. grisea-specific, and Xoo-specific. The expression patterns of representative defense-related genes were further confirmed by RT-PCR. The large-scale expression data from our microarray analysis indicated the existence of distinctive as well as shared defense pathways between the rice–M. grisea and rice–Xoo interactions.     

Pi34-AVRPi34: a new gene-for-gene interaction for partial resistance in rice to blast caused by Magnaporthe grisea

The japonica rice (Oryza sativa) cultivar Chubu 32 has a high level of partial resistance to blast, which is mainly controlled by a dominant resistance gene located on chromosome 11. The partial resistance to the rice blast fungus (Magnaporthe grisea) in Chubu 32 has isolate specificity; isolate IBOS8-1-1 is more aggressive on Chubu 32 than are other isolates. We hypothesized that the gene-for-gene relationship fits this case of a partial resistance gene in Chubu 32 against the avirulence gene in the pathogen. The partial resistance gene in Chubu 32 was mapped between DNA markers C1172 (and three other co-segregated markers) and E2021 and was designated Pi34. In the 32 F₃ lines from the cross between a chromosome segment substitution line (Pi34⁻) from Koshihikari/Kasalath and Chubu 32, the lines with high levels of partial resistance to the M. grisea isolate Y93-245c-2 corresponded to the presence of Pi34 estimated by graphic genotyping. This indicated that Pi34 has partial resistance to isolate Y93-245c-2 in compatible interactions. The 69 blast isolates from the F₁ progeny produced by the cross between Y93-245c-2 and IBOS8-1-1 were tested for aggressiveness on Chubu 32 and rice cultivar Koshihikari (Pi34⁻). The progeny segregated at a 1 : 1 ratio for strong to weak aggressiveness on Chubu 32. The results suggested that Y93-245c-2 has one gene encoding avirulence to Pi34 (AVRPi34), and IBOS8-1-1 is extremely aggressive on Chubu 32 because of the absence of AVRPi34. This is the first report of a gene-for-gene relationship between a fungal disease resistance gene associated with severity of disease and pathogen aggressiveness.     

Expression and homology modelling of sterol 14α‐demethylase of Magnaporthe grisea and its interaction with azoles

BACKGROUND: Magnaporthe grisea (Hebert) ME Barr infection is one of the most serious diseases for cultivated rice in the world. Sterol 14α‐demethylase (CYP51) is an important drug target for microbial pathogenic infections. To exploit specific and effective fungicides for M. grisea better, the authors have analysed the characteristics of interaction between sterol 14α‐demethylase from M. grisea (MGCYP51) and azoles. MGCYP51 with truncation of N‐terminal residues was cloned and expressed in E. coli, difference binding spectra of MGCYP51 induced by addition of four commercial azoles were determined and molecular modelling of MGCYP51 based on the crystal structure of Mycobacterium tuberculosis Lehmann & Newman and docking with the azoles were performed. RESULTS: The affinity of the azoles for MGCYP51 was positively correlated with their hydrophobicity. Amino acid residues Tyr112, Phe120, Phe220, His308 and Phe497 of MGCYP51, forming a large hydrophobic cavity, are the key residues interacting with azole fungicides. Furthermore, Phe220 and Phe497 are fungus and species specific respectively. CONCLUSION: The results suggest that the more potent azole fungicides for MGCYP51 should possess more hydrophobic groups interacting with residues Phe220 and Phe497. Copyright © 2009 Society of Chemical Industry     

新型含2(1H)-喹啉酮的甲氧基丙烯酸酯类

以(E)-2-[2'-(bromo-methyl)phenyl]3-甲氧基丙烯酸甲酯和2(1H)-喹啉酮类化合物为原料,通过醚化等反应,合成了11个新型含2(1H)-喹啉酮结构的甲氧基丙烯酸酯类化合物,其结构经红外、电喷雾串联质谱(ESI-MS)和核磁共振氢谱确证。初步生物活性测试结果表明:在100 mg/L质量浓度下,部分目标化合物对烟草赤星病菌Alternaria alternata等6种病原菌具有一定的抑菌活性,其中5a对水稻稻瘟病菌Pyricularia grisea的抑菌率达100%。     

稻瘟病菌对稻瘟灵抗性遗传研究

在离体条件下就稻瘟病菌对稻瘟灵抗性的诱导、抗性水平和遗传进行了研究。结果表明,供试３个小种（ＺＡ_４９、ＺＦ_１和ＺＤ_１）４个菌株分别经稻瘟灵５０μｇ／ｍｌ、１００μｇ／ｍｌ和稻瘟灵１００μｇ／ｍｌ＋亚硝基胍０.５μｇ／ｍｌ的３种处理诱变,均得到了抗稻瘟灵突变株,在含稻瘟灵的培养基中加入诱变剂亚硝基胍可显著提高稻瘟病菌对稻瘟灵抗性的突变率。上述３个不同处理获得的突变株的抗性水平相似,为野生型亲本的２.８～８.８倍。突变株对稻瘟灵的抗性在单分生孢子无性系后代可以稳定遗传。     

Suppression of rice blast by phylloplane fungi isolated from rice plants

Rice phylloplane fungi were evaluated for their potential as biocontrol agents for rice blast disease caused by Magnaporthe grisea. A total of 1923 fungal isolates were obtained from rice plants in fields at Ishigaki and Iwama and from potted plants placed in a cedar woods in Iwama as bait. Although 82.9% of isolated fungi could not be identified, species of Epicoccum were the most prevalent among identified isolates. Of the 1923 isolates, 967 were randomly selected for screening against rice leaf blast. Nine isolates (MKP5111B, MKP5112, J2JMR3-2, K2J131-2, I5R3-1, NOP541, K1KM134-1, NOP5112, MKP33222) suppressed the disease when a conidial or hyphal suspension of both the phylloplane fungus and pathogen were simultaneously used to inoculate rice plants cultured in pots in a growth chamber. Five of the isolates originated from potted plants in the woods and four from Ishigaki, a subtropical island. Five (MKP5111B, MKP5112, NOP541, NOP5112, MKP33222) of the nine isolates strongly suppressed conidial germination of M. grisea (0.7%) and formed inhibition zones (3–5mm width) in dual cultures with the pathogen. Methanol extracts from the isolates also inhibited mycelial growth of the pathogen. These results suggest that the five isolates produced antibiotic(s). These five isolates are likely identical or closely related fungal species because the sequence of their ITS regions were 100% similar. ITS sequence analysis also suggested that J2JMR3-2 was associated with a species of Fusarium. Under field conditions, J2JMR3-2 reduced both leaf and panicle blast severity, and three other isolates (MKP5111B, K1KM134-1, K2J131-2) suppressed leaf blast in one of the three experiments.     

New South Wales Department of Agriculture,Entomology Branch,Annual Report 1966-7 (extracts)

Abstract

Damage to tea plants due to a root infection was observed on an estate in southern Malawi. Initial symptoms on plants in the infected areas varied from yellowing of leaves on individual branches followed by dieback to a more frequent rapid wilting of the leaves of entire bushes. The bark surface of superficial roots and collar region was covered by a loose mat of coarse fungal mycelium varying from white or cream to pale or bright yellow in colour. Infection was well developed on a large proportion of roots of affected plants before foliage symptoms appeared. The causal agent was identified as Pseudophaeolus baudonii (Pat.) Ryv.; this is the first record of the fungus in Malawi and the first record of its occurrence anywhere on tea. The fungus is widely distributed in Africa on a range of hosts. It is not yet known how the fungus is transmitted and definite control methods cannot be recommended although it is suggested that entire tea crops in diseased areas be removed and the land used for tobacco production. It is unlikely thatP. baudonii will have serious economic effects on tea crops in the area.     

Effects of some indole-related compounds on the infection behavior of Magnaporthe grisea

Based on their effect on the infection behavior of Magnaporthe grisea, indole-related compounds were classified into three groups. The first group, including tryptophan, indole-3-butyric acid, indole-3-pyruvic acid, and indole-3-acetamide, did not inhibit infection behavior such as spore germination, appressorium formation, or infection hypha formation in M. grisea. The second, including indole acetic acid, indole-3-acetonitrile, oxindole, and tryptamine inhibited all stages of infection behaviors in a dose-dependent manner. The third, including gramine and indole, did not inhibit spore germination or appressorium formation, whereas it did inhibit infection hypha formation in a dose-dependent manner. These results suggest that endogenous or exogenously applied indole-related compounds in the second and third groups may contribute to protection in blast-susceptible plants such as rice and barley.     

First record of the tomato leafminer,Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) in Sudan

An outbreak of larvae and adults of a devastating moth was noted to be causing severe damage in tomato grown in greenhouses at Khartoum State, Sudan during June 2010. Symptoms included foliage mining and leaves drying. Photographs of symptoms and larvae and adults of the pest (and subsequently specimens) were sent to Russell IPM, UK and the Finnish Museum of Natural History for identification. The pest was identified as Tuta absoluta (Meyrick), (Lepidoptera: Gelechiidae). This is the first official record of T. absoluta in Sudan, based on surveys in tomato production areas in Khartoum, Gezira, Red Sea and North Kordofan States and eggplant and potato in Khartoum State. Severe yield loss due to the serious feeding damage on foliage and fruits infestation was reported on tomato and eggplant fields at Khartoum State.     

Transgenic indica rice expressing a bitter melon (Momordica charantia) class I chitinase gene (McCHIT1) confers enhanced resistance to Magnaporthe grisea and Rhizoctonia solani

McCHIT1 chitinase (DQ407723), a class I secretory endochitinase from bitter melon (Momordica charantia), had been demonstrated to enhance resistance against Phytophthora nicotianae and Verticillium wilt in transgenic tobacco and cotton. In order to obtain disease-resistant transgenic rice, McCHIT1 was transformed into a restorer line JinHui35 (Oryza sativa subsp. indica) by using the herbicide-resistance gene Bar as the selection marker. Transgenic rice lines and their progenies overexpressing the McCHIT1 gene showed enhanced resistance to Magnaporthe grisea (rice blast) and Rhizoctonia solani (sheath blight), two major fungal pathogens of rice. McCHIT1-transgenic rice confirmed the inheritance of the transgene and disease resistance to the subsequent generation. The T₂ transformants exhibited significantly increased tolerance to M. grisea, with a 30.0 to 85.7 reduction in disease index, and R. solani, with a 25.0 to 43.0 reduction in disease index, based on that of the control as 100. These results indicated that over-expression of the McCHIT1 gene could lead to partial disease reduction against these two important pathogens in transgenic rice.