我国水稻白叶枯病和条斑病发生与防控研究进展

培育和种植抗病品种和研发新型绿色杀菌剂是防治水稻白叶枯病和条斑病的有效措施。最近几年有关稻黄单胞菌Xanthomonas oryzae致病效应蛋白调控水稻抗(感)病性研究取得了突破性进展, 为水稻抗性品种培育提供了新思路、新策略。本文对稻黄单胞菌-水稻互作系统中已知的TALE效应蛋白与水稻抗(感)病基因(R 或 S)的对应关系进行了归纳, 就tal基因与水稻R或S基因的协同进化进行了分析, 结合生物农药和高效低毒杀菌剂的应用现状, 提出了我国水稻白叶枯病和水稻条斑病绿色防控关键策略。     

miR1320表达水平与水稻白叶枯病抗性的关联性分析

水稻白叶枯病是由稻黄单胞菌水稻致病变种Xanthomonas oryzae pv. oryzae(Xoo)引起的一种细菌性病害,严重影响水稻的产量和品质。miR1320是水稻与Xoo互作中重要的调控因子。本研究在接种Xoo后的0、12、24 h分别检测了10个粳稻和8个籼稻品种中miR1320的积累和4个防卫基因的转录水平,测量了各品种叶片上的病斑长度,分析了miR1320的积累和防卫基因的表达与水稻白叶枯病抗性之间的相关性。结果表明:接种Xoo后,miR1320的表达模式在不同品种中有所不同;籼稻和粳稻对白叶枯病的抗性分别与接种后24 h和12 h时miR1320的表达量呈紧密负相关;且相对于防卫基因,miR1320更能反映品种间的抗病性。因此,miR1320可应用于评估水稻品种对白叶枯病的抗性,辅助水稻抗病种质的鉴定。     

云南高海拔粳稻区白叶枯病菌致病型的鉴定

水稻白叶枯病是危害水稻的重要细菌性病害之一,病原为稻生黄单胞菌水稻致病变种Xanthomonas oryzae pv.oryzae (Xoo).近10年来云南省水稻白叶枯病的危害向高海拔粳稻区扩散,在海拔2200m的剑川县,白叶枯病已成为最严重的水稻病害[1-2].开展高海拔粳稻区白叶枯病菌致病型的鉴定、分布和分化研究,对云南高海拔粳稻区稻作生产具有指导意义.     

云南疣粒野生稻抗白叶枯病鉴定及叶片组织学观察

 本研究采用水稻白叶枯病菌强致病性代表菌株,评价了云南省自然分布的疣粒野生稻代表生态居群18份材料的抗病性,观察叶片组织学结构,初步分析了疣粒野生稻的抗性基因。结果表明,16份材料表现中抗、高抗甚至免疫;9份材料对4个菌株的抗性有一定差异;同时发现云南地方代表菌株X1和CN9404的致病性比C1和BD8438强。叶片组织学结构表明,高抗材料与感病的栽培稻米泉黑芒都无蜡质层,二者的叶肉组织、薄壁细胞和表皮细胞相同,表皮毛、维管束、厚壁组织等的差异也不明显,不足以引起抗病性差异。疣粒野生稻高抗或免疫白叶枯病并非借助其特殊叶片结构的物理作用,而主要是抗病基因的作用。参试疣粒野生稻中没有与Xa1、Xa21同源的基因,加之独立进化,推断其具有新的优异抗白叶枯病基因,值得深入研究和发掘利用。     

水稻品种对稻瘟病和白叶枯病抗性的遗传

选用稻瘟病(Pyricularia oryzae)生理小种ZB15和白叶枯病(Xanthomonas campesfris pv.oryzae)菌株98对3个水稻品种的抗性基因进行了遗传分析。结果表明IR2061和戈马列的稻瘟病抗性分别受非等位的2对重复显性基因所控制。IR29的抗性表现较为复杂,其抗性遗传尚待进一步研究。IR29和IR2061具有1对相同的抗白叶枯病显性基因,而戈马列则具有1对非等位的显性抗性基因,并且受珍汕97的1对隐性抑制基因所制约。     

水稻白叶枯病抗性基因在不同载体品种上及杂种F_1的抗性研究

本研究测试了八个水稻白叶枯病抗性基因Xa3、Xa4、xa5、Xa7、Xa10、Xa11、Xa14和Xa23在相应载体品种上对华南白叶枯病优势致病菌系Ⅳ型菌和强毒菌系Ⅴ型菌的抗性反应;分析了Xa4、xa5、Xa7和Xa23抗病基因与不同感病品种杂交组合F_1的抗性表达模式以及显性抗病基因Xa7和Xa23在杂交水稻上的利用价值。结果表明:大部分抗性基因在不同载体品种上抗性表达一致,但也有少数基因在不同载体品种上抗性表现不一,甚至截然相反,说明不同遗传背景对抗性表达有影响,而这种影响因不同抗性基因而异。隐性抗病基因xa5(IRBB5)和显性抗病基因Xa23(CBB23)与感病亲本组合的F_1代分别表现感病和抗病,符合隐性基因或显性基因的抗性表达模式;而2个显性抗病基因Xa4(IRBB4))和Xa7(IRBB7)与感病亲本组合的F_1代,有部分组合的F_1代表现抗病,符合显性基因的抗性表达模式,有部分组合的F_1代表现感病,不符合显性基因的抗性表达模式。在杂交水稻上的利用价值方面,Xa7与2个不育系组合的F_1代均表现高感,不宜在杂交水稻上利用;Xa23(CBB23)与4个感病亲本,无论是不育系还是常规稻组合的4个F_1代全部表现抗病,抗性在F_1充分表达,在杂交水稻上有重要的利用价值。     

水稻黄单胞菌致病相关基因插入突变体系的建立

 水稻白叶枯病菌（Xanthomonas oryzae pv. oryzae, Xoo）和条斑病菌（Xanthomonas oryzae pv. oryzicola, Xoc）是水稻上的模式病原菌,分别引起水稻白叶枯病（bacterial blight, BB）和细菌性条斑病（bacterial leaf streak, BLS）。为了精确和高效地实现目的基因的突变,本研究利用pK18mobGⅡ载体,建立了一套适于Xoo和Xoc目的基因定点插入的突变体系。通过同源片段与目的基因间的同源重组,成功获得了Xoo和Xoc的hrcV和hrpF突变体。PCR和Southern杂交证实：pK18mobGⅡ携带不同大小的同源片段,能够整合于hrcV和hrpF的特定位点;200～400 bp的同源片段能够获得最佳的突变效率;两亲接合的转化效率是电转化的5～100倍。毒性测定结果显示,hrcV基因决定着Xoo在水稻上的致病性。致病相关基因插入突变体系的建立为研究水稻黄单胞菌与水稻互作中致病相关基因的功能奠定了遗传学研究基础。     

水稻抗白叶枯病基因Xa47(t) a的敲除及抗性鉴定

为鉴定水稻Xa47(t)a基因对白叶枯病的抗性,以水稻种质L214为材料,通过构建针对Xa47(t)a基因的Xa47(t)a-Cas9敲除载体来获得敲除突变体,利用生物信息学技术对突变的Xa47(t)a基因进行突变类型分析,同时采用实时荧光定量PCR(quantitative real-time PCR,qPCR)技术分析突变体中Xa47(t)a及病程相关基因的表达情况,并于水稻孕穗期对突变体及其野生型植株接种11株白叶枯病菌Xanthomonas oryzae pv. oryzae菌株进行抗性鉴定。结果表明,在Xa47(t)a基因的第2外显子区域进行基因编辑后成功获得25株T₀代突变体株系;测序分析发现T₀代突变体株系中有13种不同的突变体类型,其中纯合突变体有3种类型,且突变位点均在靶标位点的11位碱基处缺失1～4个A碱基;氨基酸序列分析发现大部分突变体中Xa47a编码的蛋白翻译会提前终止,由原来的803个氨基酸变为144～166个氨基酸;qPCR分析结果表明突变体中Xa47(t)a及大部分病程相关基因的表达水平显著低于野生型株系;抗性鉴...     

安徽水稻白叶枯病菌生理小种监测及其对水稻主栽品种的致病性测定

水稻白叶枯病是由稻黄单胞菌水稻致病变种Xanthomonas oryzae pv.Dryzae引起的细菌性病害[1-2],该病菌具有明显的致病力差异,有关其致病力分化研究一直备受关注[2-3].水稻白叶枯病是安徽省水稻上的重要病害,自20世纪90年代承河元等[4]确定安徽省水稻白叶枯病菌中存在0、Ⅰ、Ⅱ和Ⅲ致病型以来,至今未见关于其生理小种监测的系统报道.     

22份中国小麦生产主栽品种抗条锈病基因解析

为挖掘新的小麦抗条锈病基因,掌握小麦生产主栽品种的抗条锈病基因携带情况,有效防控小麦条锈病,采用抗性鉴定、基因推导分析和分子标记技术对22份小麦生产上主栽品种进行了研究,通过抗性鉴定比较22份小麦主栽品种与已知基因载体品种的抗谱。结果显示,共推测出14份供试品种携带已知抗条锈基因,8份供试品种携带未知抗条锈基因,是新的抗锈基因资源;聚类分析结果显示,供试22份小麦品种可分为2个大类6个亚类;利用SSR分子标记检测抗条锈病基因Yr1、Yr10和Yr24的携带情况发现,11份品种携带Yr1基因,2份品种携带Yr10基因,22份品种均不携带Yr24基因。部分生产主栽品种携带新的抗条锈病基因,表明小麦品种选育中避免了抗性基因单一化,并加强了未知基因的利用。     

Influence of rice development on the function of bacterial blight resistance genes

Disease resistance genes most commonly used in breeding programs are single, dominant genes with relative effectiveness that is sometimes influenced by plant developmental stage. Knowing the developmental stages at which a resistance gene is functional is important for disease management. In rice, resistance at the seedling stage is crucial, because wounding during transplanting increases the potential for bacterial blight disease, and not all bacterial blight resistance genes are effective at the seedling stage. Effectiveness of the bacterial blight resistance genes Xa4, xa5, and Xa7, all in a common genetic background, was evaluated at different developmental stages by measuring lesion length and bacterial numbers after inoculation with the bacterial pathogen, Xanthomonas oryzae pv. oryzae. The Xa4 and xa5 genes controlled disease at all growth stages. In contrast, Xa7 was not fully functional in very young seedlings, but was completely effective by 21 days after sowing (das). The effects of plant developmental stage on interactions of the Xa7 gene with X. oryzae pv. oryzae strains carrying different mutant avrXa7 alleles were also tested. If a partial or fully functional avrXa7 allele was present, Xa7 resistance was effective at all growth stages tested after the transplant stage (>21 das).     

Genome‐wide identification of pathogenicity genes in Xanthomonas oryzae pv. oryzae by transposon mutagenesis

A transposon mutant library was constructed from the bacterial blight pathogen Xanthomonas oryzae pv. oryzae (Xoo) KACC10331 by Tn5 transposon mutagenesis. The susceptible rice cultivar Milyang 23 was inoculated with a total of 24 540 mutants resistant to kanamycin and 67 avirulent or reduced‐pathogenicity mutant strains were selected for study. Southern hybridization verified that 84 mutant strains had single‐copy insertions and their single‐transposon insertion sites were identified by sequencing analysis combined with thermal asymmetric interlaced (TAIL)‐PCR. The single‐transposon‐tagged sequences of 21 mutant strains belonged to pathogenicity‐related genes previously reported in Xanthomonas species, while the other 46 single‐transposon‐tagged sequences included diverse functional genes encoding, five cell‐wall‐degrading enzymes, three fimbrial and flagella assembly regulators, five regulatory proteins, 15 metabolic regulators and 18 hypothetical proteins, which were identified as novel pathogenicity genes of Xoo.     

Identification and molecular mapping of the rice bacterial blight resistance gene allelic to <Emphasis Type="Italic">Xa7</Emphasis> from an elite restorer line Zhenhui 084

Rice bacterial blight (BB), caused by Xanthomonas oryzae pv. Oryzae (Xoo), is a serious disease in rice production worldwide. Rice cv. Zhenhui 084, a newly developed strong indica restorer line, exhibits high resistance to most of the Philippine races of BB and has been widely used in rice hybrids in China; however, the resistance gene has not yet been cloned. Here, we show that the resistance of Zhenhui 084 to Xoo strains is similar to that of IRBB7 containing Xa7, a durable and broad resistance dominant gene for BB. To map the resistance gene in Zhenhui 084, a F₂ population with 331 highly susceptible individuals derived from a cross between Chenghui 448 and Zhenhui 084 was built. We finely mapped the target R gene to a region between two proximal markers RM20576 and MY4 in rice chromosome 6. A marker-based physical map of chromosome six was used to construct the contig covering the genomic region between two markers RM20576 and MY4. The target gene was assumed to be in an interval of approximate 200 kb, in which 16 candidate genes were predicted. Our findings will greatly facilitate the isolation and characterisation of the target R gene allelic to Xa7. Additionally, two PCR-based markers, tightly linked to the target R gene locus, will be a useful tool for the marker-assisted selection of the target R gene allelic to Xa7 in breeding programmes.     

Breeding approaches for bacterial leaf blight resistance in rice (Oryza sativa L.), current status and future directions

Bacterial leaf blight (BLB) is one of the most serious threats to the rice crop in irrigated and rainfed areas of the world. It is caused by Xanthomonas oryzae pv. oryzae and has been known for more than a century. Through rigorous screening and selection, a number of resistant cultivars have been produced and utilized, but resistance was overcome by the development of mutant strains of pathogen and by the dynamic change in Xoo populations. About 38 resistance genes have been reported in rice against the disease and a few have been cloned. The pyramiding of several resistance genes through marker assisted selection has been a quite effective strategy for combating the disease. However, new powerful tools such as transgenics have been introduced to make a significant impact. The purpose of this mini-review is to consolidate the existing knowledge about bacterial leaf blight in rice and the progress made both in conventional as well as in molecular dimensions of breeding together with potential findings and constraints.     

水旱两种栽培模式下海南山栏稻对白叶枯病抗性鉴定与评价

为研究山栏稻对白叶枯病抗性,本试验以17个山栏稻品种为材料,在大田与温室大棚中同时种植,于大田中观察山栏稻在水、旱两种栽培模式下对海口当地白叶枯病菌的自然抗性并测产;在温室中检测水、旱两种栽培模式下山栏稻对菲律宾白叶枯病菌小种PXO99(P6)、PX145(P7)的抗性;并以基因功能性标记检测其抗性基因,分析各指标与抗病性的关系。结果表明,大田环境下17个供试品种中有12个品种在水作栽培模式下感病指数高于旱作,有10个品种水作栽培模式下产量比旱作高。温室中用P6接种山栏稻,水作模式下仅有1个品种表现抗病,8个品种感病;旱作模式下2个品种表现抗病,3个品种感病。接种P7,水作时表现抗病和感病的品种数量分别为7个和2个;旱作时表现抗病的品种高达9个,仅1个品种感病。供试品种中有16个品种含有Xa1抗病基因,4个品种含有Xa27抗病基因,所有品种均不含xa13、Xa21抗病基因。因此认为,山栏稻对海口当地白叶枯病菌小种以及菲律宾白叶枯病菌小种都有一定抗性,且抗性受栽培条件的影响,山栏稻旱作时抗性强于水作。     

Virulence profiling of <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis> isolates,causing bacterial blight of rice in India

Bacterial blight (BB) of rice caused by Xanthomonas oryzae pv. oryzae (Xoo), remains a major production constraint in rice cultivation especially in irrigated and rainfed lowland ecosystems in India. The pathogen is highly dynamic in nature and knowledge on pathotype composition among the Xoo population is imperative for designing a scientific resistance breeding program. In this study, four hundred isolates of Xoo collected from diverse rice growing regions of India were analyzed for their virulence and genetic composition. Virulence profiling was carried out on a set of differentials consisting of 22 near isogenic lines (NILs) of IR24 possessing different BB resistance genes and their combinations along with the checks. It was observed that different NILs possessing single BB resistance gene were susceptible to about 59–94% of the Xoo isolates except IRBB 13 (containing BB resistance gene xa13), which showed susceptibility to about 35% of the isolates. Based on the reaction of the Xoo isolates on the differentials, they were categorized into 22 pathotypes. Among the 22 pathotypes, IXoPt-1 and IXoPt-2 were least virulent and IXoPt # 18–22 were highly virulent. Pathotype IXoPt-19 which was virulent on all single BB resistance genes except xa13 constituted the major pathotype (22.5% isolates) and was widely distributed throughout India (16 states). This was followed by pathotype IXoPt-22 (17.25%) which was virulent on all the NILs possessing single BB resistance genes. Molecular analysis was carried out using two outwardly directed primers complementary to sequence of IS1112, a repetitive element of Xoo. A high level of genetic polymorphism was detected among these isolates and the isolates were grouped into 12 major clusters. The data indicated complex nature of evolution of the Xoo pathotypes and there was no strong correlation between pathotypes and genetic clusters as each genetic cluster was composed of Xoo isolates belonging to different pathotypes. The study indicated that none of the single BB resistance genes can provide broad spectrum resistance in India. However, two-gene combinations like xa5 + xa13 and different 3 or 4 genes combination like Xa4 + xa5 + xa13, Xa4 + xa13 + Xa21, xa5 + xa13 + Xa21 and Xa4 + xa5 + xa13 + Xa21 are broadly effective throughout India.     

Identification and fine mapping of the new bacterial blight resistance gene, <Emphasis Type="Italic">Xa31(t)</Emphasis>, in rice

Zhachanglong (ZCL), a regional rice variety from Yunnan province in southwest China, has a high level of resistance to a broad spectrum of Xanthomones oryzae pv. oryzae (Xoo) isolates. In a previous study, a bacterial blight (BB) resistance (R) gene, Xa22(t), with resistance against Xoo strain Px061 on chromosome 11 was identified in ZCL. Here, we report another BB R-gene, tentatively named Xa31(t), with resistance against Xoo strain OS105 and susceptible to Px061 identified in ZCL. To determine the location of Xa31(t), 102 polymorphic RFLP markers on 12 rice chromosomes were selected for bulked segregation analysis (BSA). Twelve RFLP markers on chromosome 4 detected DNA polymorphisms between ZCL and Zhengzhu Ai (ZZA), as well as in the OS105-resistant and -susceptible bulks from F₂ populations derived from ZCL × ZZA. Genetic linkage analysis and fine mapping localised Xa31(t) within a genetic distance of 0.2 cM between two RFLP markers, G235 and C600, on the end of the long arm of chromosome 4, using two F₂ populations from the cross ZCL × ZZA and two F₃ populations, consisting of 3,311 plants with 301 F₃ random families and 3,333 plants with 303 F₃ Pxo61-susceptible families, derived from the same F₂ populations from the cross ZCL × ZZA. Using two flanking markers, G235 and C600, to screen the MH63 BAC library, the Xa31(t) locus was limited to one BAC clone with a length of about 100 kb.     

Inheritance of resistance to bacterial blight in 21 cultivars of rice

ABSTRACT Genetic analysis for resistance to bacterial blight (Xanthomonas oryzae pv. oryzae) of 21 rice (Oryza sativa L.) cultivars was carried out. These cultivars were divided into two groups based on their reactions to Philippine races of bacterial blight. Cultivars of group 1 were resistant to race 1 and those of group 2 were susceptible to race 1 but resistant to race 2. All the cultivars were crossed with TN1, which is susceptible to all the Philippine races of X. oryzae pv. oryzae. F(1) and F(2) populations of hybrids of group 1 cultivars were evaluated using race 1 and F(1) and F(2) populations of hybrids of group 2 cultivars were evaluated using race 2. All the cultivars showed monogenic inheritance of resistance. Allelic relationships of the genes were investigated by crossing these cultivars with different testers having single genes for resistance. Three cultivars have Xa4, another three have xa5, one has xa8, two have Xa3, eight have Xa10, and one has Xa4 as well as Xa10. Three cultivars have new, as yet undescribed, genes. Nep Bha Bong To has a new recessive gene for moderate resistance to races 1, 2, and 3 and resistance to race 5. This gene is designated xa26(t). Arai Raj has a dominant gene for resistance to race 2 which segregates independently of Xa10. This gene is designated as Xa27(t). Lota Sail has a recessive gene for resistance to race 2 which segregates independently of Xa10. This gene is designated as xa28(t).