离体水稻叶片划伤接种鉴定稻瘟菌的致病型

抗瘟品种的培育和抗瘟基因布局需要快速、准确、大规模地定性水稻抗源及其后代的抗瘟基因型和稻瘟菌致病型。为此, 本研究建立了水稻离体叶片划伤接种方法。该方法依据主效抗瘟基因抵抗稻瘟菌在寄主体内扩展的特点, 通过针刺在水稻叶片上造成伤口, 避免了寄主侵入抗性的干扰, 从而有利于抗扩展性的定性鉴定。作者利用活体喷雾接种、叶片无划伤接种和本研究建立的离体叶片划伤接种等3种接种方法, 在秧苗4~6叶龄期, 对菌株12-DG-68在24个水稻抗瘟单基因系上的致病反应进行了测定, 结果显示：叶片划伤接种的检测结果稳定、一致; 而叶片无划伤接种和活体喷雾接种的检测结果假抗性比例分别为12.5%和4.2%, 不同叶龄期的叶片间反应型不一致率达7%。此外, 离体叶片划伤接种还可利用菌丝块接种, 以鉴定分生孢子产量低的菌株的致病型。因此, 水稻叶片划伤接种是一种准确、稳定和方便的稻瘟菌接种方法, 可用于大规模定性测定水稻抗源及其后代的抗瘟基因型和稻瘟菌的致病型。     

三环唑浸秧防治稻叶瘟技术研究

用20％三环唑(tricyclazole)可湿性粉剂750—1000倍液浸湿秧苗半分钟左右取出,堆置半小时后移栽,药后33—55天防效达85％以上,本田不再施药即可有效地控制本田叶瘟。较常规喷雾更有效地发挥了三环唑内吸性强、持效期长、耐雨水冲刷的特点,减少本田施药1—2次,省工省药且方法简便易行。通过四川大面积示范推广,效益非常显著,为稻瘟病综合防治提供了一项有效措施。     

Induction of systemic resistance byPseudomonas fluorescens Pf1 againstXanthomonas oryzae pv.Oryzae in rice leaves

When lower leaves of rice plants were inoculated with powder formulation of a saprophytic strain ofPseudomonas fluorescens, Pfl, upper leaves, in addition to the inoculated lower leaves, showed resistance to the rice bacterial blight pathogenXanthomonas oryzae pv.oryzae. When the leaves were challenge-inoculated withX. oryzae pv.oryzae 4 days afterP. fluorescens application on lower leaves, the disease intensity in upper leaves decreased from 6.7 to 1.1. When rice seeds were treated with the formulation ofP. fluorescens Pfl and sown, 30-day-old seedlings showed resistance toX. oryzae pv.oryzae and the disease intensity decreased from 6.8 to 1.2. The induced resistance was transient; leaves sprayed withP. fluorescens Pfl at 30 days after treatment and leaves of 60-day-old seedlings fromP. fluorescens-treated seeds did not show resistance to the pathogen. In field trials, seed treatment followed by foliar application of the powder formulation ofP. fluorescens Pfl effectively controlled rice bacterial blight and increased the yield. In the induced resistant leaves a sharp increase in lignification and activities of peroxidase, phenylalanine ammonia-lyase and 4-coumarate: CoA ligase was observed when the leaves were challenge-inoculated withX. oryzae pv.oryzae. An approximately threefold increase in lignin content, peroxidase activity and phenylalanine ammonia-lyase activity and a fivefold increase in 4-coumarate: CoA ligase activity were observed 5 days after challenge inoculation withX. oryzae pv.oryzae in rice leaves pretreated withP. fluorescens for 5 days. A similar increase in defense-related activities was not observed in susceptible interactions or inP. fluorescens-treated plants at later stages of interactions when no resistance to the pathogen was observed.     

叶枯唑防治水稻白叶枯病的作用方式研究

叶枯唑具有诱导受体植物水稻产生抗病性的作用 ,能使水稻体内脂质过氧化程度加强 ,刺激水稻体内产生 O-2 ,阻止白叶枯病菌侵入。O-2 清除剂 (甘露醇、抗坏血酸 )处理水稻能显著降低叶枯唑的保护作用。叶枯唑也能直接作用于水稻白叶枯病菌 ,抑制菌体生长 ,表现出治疗作用。     

Rice Diseases in Nigeria

Abstract

The two major fungal diseases of rice in Nigeria are rice blast (Pyricularia oryzae Cav.) and brown spot (Cochliobolus miyabeanus (Ito and Kuribayashi) Dreschler ex Dastur). The loss in grain yield attributable to these diseases ranges from 11.5–39.6% and 12–43% respectively, Mancozeb, blasticidin-S, fentin hydroxide, benomyl and edifenphos as foliar sprays effectively control rice blast in the laboratory and field. Brown spot is controlled by spraying with thiram, mancozeb or blasticidin-S. A more useful approach to the control of these diseases being adopted in Nigeria is the search for horizontally resistant varieties. Other fungal diseases of rice in Nigeria are the green smut (Ustilaginoidea virens (Cooke) Tak.), basal sheath rot (Rhizoctonia solani Kühn), sheath blight (Corticium sasakii (Shirai), Matsumoto.), leaf scald (Rhynchosporium oryzae Hashioka and Yokogi), narrow brown leaf spot (Cercospora oryzae Miyake) and bakanae disease, a foot rot caused by Fusarium moniliforme Sheldon. Two suspected cases of virus diseases have been reported; grassy stunt disease (transmitted by Nilaparvata lugens Stal.), and orange leaf disease (transmitted by Inazuma dorsalis (Motschulsky). No bacterial disease of rice has been found in Nigeria.     

Inhibition of appressorial adhesion of Pyricularia oryzae to barley leaves by fungicides

The effects of rice blast fungicides known to inhibit melanisation or penetration on the adhesion of Pyricularia oryzae to the leaf surface of barley were investigated. Adhering appressoria were counted after shaking inoculated leaves vigorously with water. On untreated leaves, appressoria began to adhere at the time when appressorial melanisation was initiated. The number of adhering appressoria then increased gradually, and most were melanised. On chlobenthiazone-treated leaves, appressoria were not melanised and were easily detached from the surface. Similar results were obtained with tricyclazole. However, after treatment with tetrachlorophthalide or pentachlorobenzyl alcohol, which are also known to prevent penetration of P. oryzae, melanised appressoria still remained after shaking in water. It is suggested that appressorial melanisation in P. oryzae is involved in appressorial adhesion. The antipenetrant action of the melanin biosynthesis inhibitors chlobenthiazone and tricyclazole may be ascribed to the lack of adhesive intensity necessary to support the mechanical shearing of the cuticle of rice plants.     

稻瘟病抗性分子机制及抗病育种策略研究进展

由稻瘟菌侵染引起的稻瘟病是威胁水稻安全生产最严重的真菌病害之一。鉴定克隆水稻抗稻瘟病基因, 系统深入研究稻瘟菌与水稻的相互作用, 揭示水稻的抗病机制, 进而创制推广抗稻瘟病新材料, 对确保粮食安全具有重要意义。本文总结了近10年来稻瘟病抗病基因和感病基因的鉴定、分子机理解析和应用等进展, 总结归纳了抗病基因聚合、分子设计育种、感病基因编辑、抗病基因的病原诱导表达等抗病育种主要策略。最后提出充分利用种质资源, 利用新技术挖掘新基因及创制新材料, 深入研究叶瘟和穗瘟抗病机制差异等是稻瘟病抗病育种下一步重点研究方向和新挑战。     

The role of silicon in preventing appressorial penetration by the rice blast fungus

To test the hypothesis that silicon (Si) confers resistance against appressorial penetration of the rice blast fungus, the proportion of appressorial penetration into the leaf epidermis to total appressoria formed was compared among rice plants amended with various rates of silica gel to those plants nonamended. The amounts of Si in the youngest leaves were consistent with the amounts of silica gel applied to the rice plants. Relative Si levels on the adaxial surface of leaves as detected by energy dispersive X-ray analysis also increased with the amounts of silica gel applied. Based on light microscopic observation of the adaxial surface of rice leaves, the proportion of appressorial penetration was reduced by increasing amounts of silica gel applied and increased with the length of period after spray inoculation. Consequently, these results strongly support the hypothesis and suggest that Si in the leaf epidermis may confer resistance against appressorial penetration. Meanwhile, the number of lesions per leaf also decreased with the amount of Si applied, while only a certain part of penetrated appressoria could become sporulating susceptible lesions. This suggests that Si also confers physiological resistance against blast infection after the penetration.     

Suppression of Rice Blast by Preinoculation with Avirulent Pyricularia oryzae and the Nonrice Pathogen Bipolaris sorokiniana

ABSTRACT Avirulent isolates of Pyricularia oryzae and isolates of Bipolaris sorokiniana, a nonrice pathogen, were used to suppress rice blast caused by P. oryzae. In greenhouse experiments, both fungi substantially reduced leaf blast when applied 24 h or more before the pathogen. B. sorokiniana, but not avirulent isolates of P. oryzae, systemically reduced disease in leaf 5 when applied to whole plants at the four-leaf stage. In field experiments, both fungi were able to reduce neck blast significantly. No increase in grain yield was obtained by using avirulent isolates of P. oryzae, whereas five sprays with B. sorokiniana from seedling to heading stages increased the grain yield in two of three experiments conducted at two locations in Nepal. The significant increase in yield was observed under high inoculum pressure of P. oryzae. Induced resistance is suggested to be involved in the suppression of disease.     

Assessment of blast disease resistance in transgenic PRms rice using a gfp-expressing Magnaporthe oryzae strain

Rice blast caused by the fungus Magnaporthe oryzae (anamorph Pyricularia grisea ) is one of the most devastating diseases of cultivated rice worldwide. In this study, a green fluorescent protein ( gfp )-expressing M. oryzae strain was generated and used to investigate the infection process in a commercial rice cultivar. Expression of the gfp gene did not affect the pathogenicity of the M. oryzae transformants. Confocal microscopy allowed in vivo imaging of this pathogen during infection of rice tissues. Magnaporthe oryzae pathogenicity was examined on both leaf and root tissues. In roots of wild-type plants, the fungus penetrated into epidermal and cortical cells, and colonized the central cylinder and xylem vessels. However, the dimorphic growth pattern typically observed during the biotrophic and necrotrophic stages of leaf colonization was not observed during colonization of root tissues. Furthermore, events occurring during infection of rice plants constitutively expressing the maize pathogenesis-related PRms gene were characterized and compared with those occurring during the interaction of this pathogen with untransformed rice plants. Fungal penetration was drastically reduced and delayed in tissues of PRms plants compared to untransformed plants. These results indicated that the gfp -expressing M. oryzae represents a strategic tool for the assessment of blast disease resistance in transgenic rice which can be also applied to the analysis of the M. oryzae interaction with other cultivars or mutants of important crop species.     

Effects of Elevated Atmospheric CO(2) Concentration on the Infection of Rice Blast and Sheath Blight

ABSTRACT The effect of elevated atmospheric CO(2) concentration on rice blast and sheath blight disease severity was studied in the field in northern Japan for 3 years. With free-air CO(2) enrichment (FACE), rice plants were grown in ambient and elevated ( approximately 200 to 280 mumol mol(-1) above ambient) CO(2) concentrations, and were artificially inoculated with consist of Magnaporthe oryzae. Rice plants grown in an elevated CO(2) concentration were more susceptible to leaf blast than those in ambient CO(2) as indicated by the increased number of leaf blast lesions. Plants grown under elevated CO(2) concentration had lower leaf silicon content, which may have contributed to the increased susceptibility to leaf blast under elevated CO(2) concentrations. In contrast to leaf blast, panicle blast severity was unchanged by the CO(2) enrichment under artificial inoculation, whereas it was slightly but significantly higher under elevated CO(2) concentrations in a spontaneous rice blast epidemic. For naturally occurring epidemics of the sheath blight development in rice plants, the percentage of diseased plants was higher under elevated as opposed to ambient CO(2) concentrations. However, the average height of lesions above the soil surface was similar between the treatments. One hypothesis is that the higher number of tillers observed under elevated CO(2) concentrations may have increased the chance for fungal sclerotia to adhere to the leaf sheath at the water surface. Consequently, the potential risks for infection of leaf blast and epidemics of sheath blight would increase in rice grown under elevated CO(2) concentration.     

AtEDR1介导拟南芥对稻瘟菌的抗性(英文)

 稻瘟菌(Magnaporthe oryzae)是一种半腐生病原菌,该病原菌可以侵染水稻,产生稻瘟病。对于水稻与稻瘟菌的互作机制,人们已经进行了大量的研究。然而,鉴于水稻基因组相对较大且稻瘟菌生理小种众多,致使研究进展缓慢。本研究通过将稻瘟菌生理小种Y34侵染拟南芥生态型Col-0,发现Y34可以感染Col-0,从而建立Y34-拟南芥互作模型,用以探究Y34与抗白粉突变体edr1的关系。结果显示,edr1比Col-0对于稻瘟菌更敏感。之前有研究表明,降低SA含量的突变体(pad4,sid2,nim1)均可抑制edr1相关表型。接种Y34于edr1与pad4、sid2、nim1形成的双基因突变体发现,所有双突变体的感病性比edr1单突均明显降低。推测EDR1可能在拟南芥抗稻瘟菌方面起一定正调控作用,且其作用依赖于SA途径的相关基因。     

水杨酸诱导水稻幼苗抗瘟性的生化机制

 以0.01mM水杨酸(SA)喷雾处理或稻瘟菌孢子悬浮液接种稻苗后,叶片中苯丙氨酸解氨酶(PAL)和定位于细胞不同部位的过氧化物酶(POD),特别是与细胞壁结合的POD和细胞间隙POD的活性迅速升高,但没有明显的新POD同功酶带出现。同时叶片中木质素含量迅速增加。在SA处理后的稻叶中提取到能抑制稻瘟菌分生孢子萌发、含有Momilactone A的抑菌物质,并检测到分子量分别为48、52、59、106.5KDa的碱性病程相关蛋白(PRs)。这些生化指标的时序变化与SA诱导抗瘟性的表现相吻合。对SA诱导水稻幼苗抗瘟性的可能生化机理作了归纳。     

生防菌解淀粉芽胞杆菌Jt84干悬浮剂加工及田间评价

解淀粉芽胞杆菌Jt84菌株对水稻真菌病害具有良好的防治效果。本研究通过喷雾干燥技术，开发一种生防菌新剂型杀菌剂。采用Plackett-Burman试验及Box-Behnken试验，对解淀粉芽胞杆菌Jt84干悬浮剂喷雾干燥工艺进行了优化。结合生产实际确定干悬浮剂制备条件分别为：进风温度118.9℃，进样速率25.7 r/min（约为680 mL/h），热风流量28.56 M³/h，压力0.2 MPa，出风温度80℃，喷头口径0.7 mm。经喷雾干燥试验验证该理论预测值与实际值无显著差异。田间药效试验表明：水稻破口期喷施解淀粉芽胞杆菌Jt84干悬浮剂75 g/667 m²，对水稻穗颈瘟的防治效果为79.3%，防治效果与三环唑相当，能够有效防治稻瘟病。     

Identification of specific fragments of HpaG Xooc, a harpin from Xanthomonas oryzae pv. oryzicola, that induce disease resistance and enhance growth in plants

Harpin proteins from gram-negative plant-pathogenic bacteria can stimulate hypersensitive cell death (HCD) and pathogen defense as well as enhance growth in plants. Two of these diverse activities clearly are beneficial and may depend on particular functional regions of the proteins. Identification of beneficial and deleterious regions might facilitate the beneficial use of harpin-related proteins on crops without causing negative effects like cell death. Here, we report the identification and testing of nine functional fragments of HpaG(Xooc), a 137-amino-acid harpin protein from Xanthomonas oryzae pv. oryzicola, the pathogen that causes bacterial leaf streak of rice. Polymerase chain reaction-based mutagenesis generated nine proteinaceous fragments of HpaG(Xooc); these caused different responses following their application to Nicotiana tabacum (tobacco) and Oryza sativa (rice). Fragment HpaG62-137, which spans the indicated amino acid residues of the HpaG, induced more intense HCD; in contrast, HpaG10-42 did not cause evident cell death in tobacco. However, both fragments stimulated stronger defense responses and enhanced more growth in rice than the full-length parent protein, HpaG(Xooc). Of the nine fragments, the parent protein and one deletion mutant of HpaG(Xooc) tested, HpaG10-42, stimulated higher levels of rice growth and resulted in greater levels of resistance to X. oryzae pv. oryzae and Magnaporthe grisea. These pathogens cause bacterial leaf blight and rice blast, respectively, the two most important diseases of rice world-wide. HpaG10-42 was more active than HpaG(Xooc) in inducing expression of several genes that regulate rice defense and growth processes and activating certain signaling pathways, which may explain the greater beneficial effects observed from treatment with that fragment. Overall, our results suggest that HpaG10-42 holds promise for practical agricultural use to induce disease resistance and enhance growth of rice.     

稻瘟病生防芽胞杆菌的筛选及防治效果

筛选稻瘟病生防芽胞杆菌,为水稻可持续发展中稻瘟病的防治提供生防种质资源。通过平板对峙培养和离体叶片法筛选稻瘟病菌的拮抗细菌,通过形态特征观察、biolog和16S rDNA序列分析法进行鉴定,田间试验明确供试菌株对稻瘟病的防治效果和促生作用。从280株分离菌中筛选获得3株拮抗细菌对稻瘟病菌具有较好的抑制作用,在番茄燕麦培养基平板上与稻瘟病菌Magnaporthe oryzae P131对峙培养2 d后,菌株SYX04和SYX20的菌丝生长抑制率分别是95.6%和97.5%,菌株S?22菌丝生长抑制率是75%;离体叶片法明确SYX04和SYX20防治效果分别是89.6%和91.9%,与20%三环唑溶液没有显著差异,菌株S?22的防治效果是80.1%。根据16S rDNA序列分析比对结果,菌株SYX04和SYX20与枯草芽胞杆菌属基因序列的同源性最高,结合形态观察和biolog鉴定结果,确定这2株菌为枯草芽胞杆菌Bacillus subtilis,菌株S?22鉴定为多食鞘氨醇杆菌Sphingobacterium multivorum。田间试验结果表明,菌株SYX04和SYX20对水稻有较好的促生作用,分别促进水稻根长6.7%和13.2%、增加株高7.8%和8.2%、增加穗重均为23.1%、增加千粒重分别达到10.7%和11.1%。菌株SYX20对叶瘟的田间防效是73.5%～83.5%,对穗瘟的防治效果是64.0%～85.6%,与75%三环唑可湿性粉剂之间没有显著差异。菌株SYX04对叶瘟的防治效果是71.5%～82.6%,对穗瘟的防治效果是63.5%～83.8%,与绿地康处理之间没有显著差异,但与75%三环唑可湿性粉剂有差异。菌株S?22对叶瘟的防治效果是69.2%～73.3%,对穗瘟的防治效果是61.6%～77.9%。     

In vitro and in vivo antifungal activities of decursin and decursinol angelate isolated from Angelica gigas against Magnaporthe oryzae, the causal agent of rice blast

Blast is considered the most important fungal disease of rice due to its wide distribution and destructiveness under favorable conditions. Development of new effective and environmentally benign agents against the causal pathogen, Magnaporthe oryzae, is of great interest. In the course of a search for natural antifungal compounds in medicinal plants, we found that the methanol extract of Angelica gigas roots showed a potent control efficacy against rice blast caused by M. oryzae. We isolated antifungal coumarins from the extract, and they were identified as decursin and decursinol angelate. Antifungal activities of these compounds, along with kasugamycin, were tested on M. oryzae in vivo and in vitro. In an in vivo assay, the three compounds effectively suppressed the development of rice blast at concentrations more than 100 μg/mL. Coumarins showed relatively weak inhibitory effect on fungal mycelial growth when compared to kasugamycin. However, they strongly inhibited M. oryzae spore germination, which was not observed in kasugamycin treatments. This is the first report demonstrating that decursinol angelate can provide control against rice blast and that the two coumarins inhibit M. oryzae spore germination. In addition, the wettable powder formulation of the crude extract of A. gigas prohibited the development of blast symptoms on rice plants more effectively than liquid concentrate formulation of kasugamin, a commercial fungicide. Based on our study, we propose that coumarin compounds as well as the A. gigas root crude extract can be used as natural, benign fungicides for controlling rice blast.     

Magnaporthe oryzae conidia on basal wheat leaves as a potential source of wheat blast inoculum

Wheat blast caused by Magnaporthe oryzae Triticum causes significant losses on wheat during outbreak years in several South American countries. Despite reports of wheat blast leaf lesions on some wheat cultivars, the importance of inoculum originating from leaves in severely affected commercial fields is disputed. It is generally considered that leaf lesions and/or sporulation on leaves do not usually appear before the occurrence of spike blast in wheat. The purpose of this study was to (i) determine the occurrence of wheat blast on basal leaves, (ii) estimate the number of conidia produced on these leaves, and (iii) determine the impact of current fungicide application practices on inoculum produced from sporulating lesions on basal wheat leaves. Inoculations at the three‐leaf stage showed that certain cultivar and isolate combinations caused more disease on old wheat leaves than young expanding leaves. Under optimum conditions, M. oryzae had the potential to produce tens to hundreds of thousands of conidia on small amounts of wheat basal leaves. A mean of 1 669 000 conidia were produced on 1 g dry basal leaves of a highly susceptible cultivar under optimum conditions for sporulation. Conidia production on leaves coincided with spike emergence under both greenhouse and field conditions. When field studies were conducted under natural epidemic conditions, foliar fungicide applications reduced the amount of M. oryzae conidia on basal leaves by 62–77% compared to non‐sprayed controls. An earlier application of foliar fungicides might reduce inoculum if conidia from basal leaves contribute to wheat spike blast development.     

利用绿色荧光蛋白GFP研究稻瘟病菌与水稻的互作

稻瘟病菌Magnaporthe oryzae严重威胁水稻的产量与质量,明确稻瘟病菌与水稻互作过程及机理,对防治稻瘟病具有重要意义。本研究利用稻瘟病菌常用致病菌株GUY11和ZB25,构建了绿色荧光蛋白GFP的过量表达菌株,并通过荧光显微观察菌株侵染寄主水稻过程中侵染结构的形成与发育,包括孢子萌发、附着胞形成、侵染钉形成、侵染菌丝增殖、坏死斑形成及产孢。另外,通过比较过量表达菌株对稻瘟病高抗水稻和易感水稻的侵染过程,发现侵染过程的差异主要集中于侵染钉的穿透和侵染菌丝的定殖。本研究为分析稻瘟病菌对寄主水稻的定殖规律提供了一种有效工具。     

28%三环唑·嘧菌酯悬浮剂对水稻稻瘟病的防治效果

为科学合理地选择和使用田间控制稻瘟病的杀菌剂,本研究通过室内苗期测定和大田试验,评价复配剂28%三环唑·嘧菌酯SC对水稻稻瘟病的防治效果。结果表明,28%三环唑·嘧菌酯SC对水稻苗瘟及穗瘟均有很好的预防和治疗效果。在温室人工控制条件下,施药后接种的防效高于接种后施药的防效,最高防效分别为97.35%和92.70%。在田间对穗瘟的最高防效达到91.11%,与对照药剂75%三环唑WP和250 g/L嘧菌酯SC的平均防效相当,可以在生产上推广使用,推荐有效成分用量为252～420 g/hm~2。