Natural Variation at the Pi-ta Rice Blast Resistance Locus

ABSTRACT The resistance gene Pi-ta protects rice crops against the fungal pathogen Magnaporthe grisea expressing the avirulence gene AVR-Pita in a gene-for-gene manner. Pi-ta, originally introgressed into japonica rice from indica origin, was previously isolated by positional cloning. In this study, we report the nucleotide sequence of a 5,113-base pair region containing a japonica susceptibility pi-ta allele, which has overall 99.6% nucleotide identity to the indica Pi-ta allele conferring resistance. The intron region shows the levels of sequence diversity that typically differentiate genes from indica and japonica rices, but the other gene regions show less diversity. Sequences of the Pi-ta allele from resistant cultivars Katy and Drew from the southern United States are identical to the resistance Pi-ta sequence. Sequences from susceptible cultivars El Paso 144 and Cica 9 from Latin America define a third susceptibility haplotype. This brings the total number of Pi-ta haplotypes identified to four, including the resistance allele and three susceptibility alleles. The Pi-ta locus shows low levels of DNA polymorphism compared with other analyzed R genes. Understanding the natural diversity at the Pi-ta locus is important for designing specific markers for incorporation of this R gene into rice-breeding programs.     

Allelism of rice blast resistance genes in two Chinese rice cultivars, and identification of two new resistance genes

Blast, caused by Pyricularia grisea , is a major constraint on rice production. To broaden genetic diversity for resistance to this disease, two rice cultivars, GA20 and GA25 from Yunnan Province, China, were analysed for the genetic basis of their high resistance to blast. GA20 was crossed with 10 Japanese differential cultivars, and GA25 was crossed with nine of them and with the susceptible Chinese cultivar Lijiangxintuanheigu (LTH). The resistance of GA20 was governed by two dominant genes allelic to genes at the Pi-k and Pi-ta loci. The allele at the Pi-k locus was new, based on a reaction pattern different from known alleles at this locus. It could not be shown whether or not the allele at the Pi-ta locus is new, because races with virulence for Pi-ta were not tested. GA25 has one resistance gene, which is not allelic to genes at the loci Pi-a , Pi-k , Pi-z , Pi-ta , Pi-b , and Pi-t , but is linked to the Pi-i gene on chromosome 9 with a recombination frequency of 15.1 ± 2.8%.
The new allele at the Pi-k locus in GA20 is designated as Pi-k^g (t), and the new resistance gene in GA25 as Pi15(t) .     

New gene for bacterial blight resistance in rice located on chromosome 12 identified from minghui 63, an elite restorer line

ABSTRACT Bacterial blight, caused by Xanthomonas oryzae pv. oryzae, is a serious disease of rice worldwide. A new dominant gene for bacterial blight resistance in rice, Xa25(t), was identified from Minghui 63, a restorer line for a number of rice hybrids that are widely cultivated in China. This gene conferred resistance to Philippine race 9 (PXO339) of X. oryzae pv. oryzae in both seedling and adult stages. It was mapped to the centromeric region of chromosome 12, 2.5 cM from a disease resistance gene-homologous sequence, NBS109, and 7.3 cM from a restriction fragment length polymorphism marker, G1314. The genomic location of this gene is similar to the previously identified blast resistance genes, Pi-ta and Pi-ta2.     

Identification of a new resistance gene to a chinese blast fungus isolate in the Japanese rice cultivar aichi asahi

ABSTRACT Genetic analysis of the rice cultivar Aichi Asahi and some other Japanese cultivars for the high resistance to the blast fungus isolate CHNOS58-3-1 from China was performed. All the Japanese differential cultivars were resistant to the isolate except for 'Pi No. 4', which showed moderate resistance. Analysis of the F(2) population of a cross of the susceptible cultivar Reiho and the resistant cultivar Aichi Asahi indicated that the resistance of 'Aichi Asahi' to the isolate was conferred by one dominant gene. To identify the gene in other Japanese differential cultivars, AK lines, which were derived from a cross of 'Aichi Asahi' x 'K59' and assumed to harbor no known genes except for the new one, were used for the allelism tests. The new, completely dominant resistance gene was detected in 14 differential cultivars, but not in 'Pi No. 4', 'Yashiro-mochi', and 'K1', and was designated as Pi19(t). Pi19(t) was allelic or closely linked to Pita(2) on chromosome 12. Pi19(t) was extensively distributed among Japanese traditional local cultivars.     

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。     

沿黄稻区水稻品种对水稻条纹叶枯病抗性评价

2007年和2008年在田间自然发病条件下, 鉴定了河南省沿黄稻区25个主要推广品种、育种高代品系对水稻条纹叶枯病的抗性。结果表明,在两个年份均表现为抗病的有5个粳稻（5/17）、3个籼稻（3/3）和2个旱稻（2/5）品种;但有4个品种（4/25）表现不一致。2007年17个粳稻品种中的11个表现为感病或中感（64.70%）,5个旱稻品种中2个表现感病或中感（40％）。2008年17个品种中有10个表现为感病或中感（58.80%）,5个旱稻品种中3个表现为感病或中感（60％）。 2008年对25个品种的病情变化和田间灰飞虱种群动态的调查结果表明,抗、感品种间的病情变化具有明显的差异,不同水稻品种间灰飞虱的虫口数量也具有显著的差异,但品种对水稻条纹叶枯病的抗性程度和灰飞虱的虫口数量并不完全一致。     

Identification of an Avirulence Gene in the Fungus Magnaporthe grisea Corresponding to a Resistance Gene at the Pik Locus

ABSTRACT A rice isolate of Magnaporthe grisea collected from China was avirulent on rice cvs. Hattan 3 and 13 other Japanese rice cultivars. The rice cv. Hattan 3 is susceptible to almost all Japanese blast fungus isolates from rice. The genetic basis of avirulence in the Chinese isolate on Japanese rice cultivars was studied using a cross between the Chinese isolate and a laboratory isolate. The segregation of avirulence or virulence was studied in 185 progeny from the cross, and monogenic control was demonstrated for avirulence to the 14 rice cultivars. The resistance gene that corresponds to the avirulence gene (Avr-Hattan 3) is thought to be located at the Pik locus. Resistance and susceptibility in response to the Chinese isolate in F(3) lines of a cross of resistant and susceptible rice cultivars were very similar to the Pik tester isolate, Ken54-20. Random amplified polymorphic DNA markers and restriction fragment length polymorphism markers from genetic maps of the fungus were used to construct a partial genetic map of Avr-Hattan 3. We obtained several flanking markers and one co-segregated marker of Avr-Hattan 3 in the 144 mapping population.     

小麦田看麦娘对甲基二磺隆的抗性水平及分子机制初探

看麦娘是中国长江中下游地区稻茬麦田的主要恶性杂草之一,甲基二磺隆是防治小麦田看麦娘等禾本科杂草的重要除草剂.该研究团队前期在安徽省凤台县小麦田采集到疑似抗性种群看麦娘(AHFT-01),为明确其对甲基二磺隆的抗性发生情况及潜在的抗性机制,采用温室盆栽法在整株水平上测定了该种群对甲基二磺隆及其他乙酰乳酸合成酶(ALS)抑...     

西瓜抗小西葫芦黄花叶病毒基因的连锁分子标记研究

 小西葫芦黄花叶病毒中国株系(Zucchini yellow mosaic virus Chinese strain,ZYMV-CH)是危害我国西瓜的主要病毒。本实验以抗病毒病西瓜野生种质P.I.595203与感病的普通西瓜自交系98R为亲本,采用单粒传方式得到109个E代株系,分别对亲本、F₁及109个F₃代株系群体进行了苗期抗ZYMV-CH接种鉴定,通过F₃代群体的抗感分离情况,推测得到F₂代各单株的基因型,采用集团分离分析法(bulked segregant analysis,BSA)在F₂代建立抗感基因池,以亲本、F₁和抗感基因池为模板,对640条RAPD引物进行PCR扩增筛选,其中引物AK13在亲本、F₁和抗感基因池之间扩增出一条多态性片段(644bp),在F₂代群体上验证该多态性条带与ZYMV-CH的抗性基因呈现连锁关系,遗传连锁距离为8cM,定名为AK13_-644,该连锁标记在ZYMV-CH抗性转育后代自交系上得到了验证。最终将此RAPD标记成功转化成SCAR标记SCAK13_-644,该标记可以作为西瓜抗病毒病辅助选择的分子标记。     

亚麻品系9801-1抗白粉病基因的RAPD标记

 F2 populations were obtained from the cross between 9801-1 and DIANE.Bulked segregate and RAPD analyses were employed to identify molecules linked to the resistance to powdery mildew.OPP02 amplified about 792 bp polymorphic band in all individuals from 9801-1 and resistant bulk,but absent in all individuals from DIANE and susceptible bulk.By further analysis in F2 segregating population,the polymorphic band was found to be cosegregated with the resistant gene possibly.The fragment was sequenced,     

玉米抗粗缩病遗传及分子标记研究

Maize rough dwarf disease caused by Rice black-streaked dwarf virus (RBSDV) is transmitted by planthopper in China. Identification and development of resistant hybrids are complicated because of the inconsistencies in viral disease pressure every year. Marker-assisted selection can provide means for main-taining virus resistance alleles even in the absence of disease. In this paper a F₂ segregation population was constructed to identity the molecular markers linked to the resistance gene using a cross between a resistant and a susceptible parents (Qi319×Ye107). Fifteen-day-old seedlings of F₂ population were exposed to small brown planthoppers carrying RBSDV for 3 days in specific inoculation chamber. The inoculated plants were transplanted to screenhouse after removing the insects completely. In plant maturity stage the disease resistance of all the individuals were visually assessed. The results showed that 17, 8, 11, 51 and 122 plants were scaled from 0-4 respectively, in which 0 means no symptoms and 4 represents highly susceptible. Chi-square test demonstrated that the segregation ratio of phenotype was 1∶15 (resistant: susceptible) or 1∶6∶9 (resistant∶moderate∶susceptible) in the F₂ population, indicating RBSDV resistance of maize was controlled by two recessive genes. The F₂ individuals DNA were extracted and 261 SSR (simple sequence repeat) primers derived from maize genome ten chromosomes were selected from maize GDB database to construct genetic linkage map. The linkage map consisted of 71 polymorphic SSR markers, spanning a genetic distance of 996.6 cM with an average interval of 14.0 cM between adjacent markers. The resistant and susceptible gene pools were set up for BSA (bulked segregant analysis) and 6 polymorphism markers were obtained with BSA-SSR method between the two pools. The F₂individuals were further analyzed with 6 polymorphism markers. Chi-square test showed that phi 051, umc1407 and umc1432, mapped on chromosome 7 and 10, exhibited segregation distortion significantly and very significantly in susceptible individuals. These three SSR markers were identified as potential markers linked to the resistant loci.     

Virulence structure of theMagnaporthe grisea rice population from the northwestern Himalayas

The virulence structure of theMagnaporthe grisea rice population from the northwestern Himalayan region of India was deciphered on 24 rice genotypes harboring different blast resistance genes. Matching virulences appropriate to all the rice genotypes, except Fukunishiki (Pi-z, Pi-sh) and Zenith (Pi-z, Pi-a, Pi-i), were present in the pathogen population. Moreover, a very low percentage of isolates were virulent on Tetep (Pi-ta, Pi-k ^h, Pi-4^b) and Tadukan (Pi-ta/Pi-ta ²). Although virulence was recorded on most of the lines tested, none was susceptible to all of the isolates. Three pairs of genotypes, namely, C101LAC:C101A51; K-1: Dular; and Dular: HPU-741, exhibited complementary resistance spectra as no isolate combined virulence to both the members of each of the three pairs of genotypes despite the fact that individual members were susceptible to a major portion of the pathogen population. The blast resistance genesPi-z, Pi-k ^h, Pi-l andPi-2 and their various combinations were construed to provide broad spectrum and durable blast resistance in Himachal Pradesh. Pathotype analysis revealed the existence of extremely high pathotypic diversity in the pathogen population. Based on the observed population structure forM. grisea, it was not possible to designate a minimum set of pathogen isolates that could be used in blast resistance screens to identify effective sources of blast resistance. The overall results suggested that the pathotype analysis alone is insufficient to describe the existing pathogenic variability, especially when this information has to be used for guiding the breeding programs aimed at developing durable blast resistance. However, population genetics approach of studying pathogenic specialization by monitoring the frequency of individual virulence genes and analyzing virulence gene combinations for their association or dissociation might generate useful information for developing durable blast resistance. http://www.phytoparasitica.org posting May 14, 2006.     

Novel septoria speckled leaf blotch resistance loci in a barley doubled-haploid population

The genetics of resistance to Septoria speckled leaf blotch (SSLB), caused by Septoria passerinii, was studied in the Leger × CIho9831 barley doubled-haploid population. The 140 lines in the population segregated as 102 resistant and 38 susceptible, approximating a 3:1 ratio. A recombination map was developed using diversity arrays technology and other molecular markers. Quantitative trait locus (QTL) analysis demonstrated that resistance is primarily conferred either by having the CIho9831 allele at a QTL on 6HS or by having the CIho9831 allele at both of two QTLs on 3H and 2HL. In addition, ≈1/16 of the lines were resistant for unidentified reasons. This model predicts a resistant/susceptible ratio of 11:5, which fits the phenotypic observations. Minor QTLs were detected on 2HS and 1H. DNA sequences of linked markers suggest that the 6HS, 3H, and 2HS QTLs are part of resistance gene clusters and that the 6HS and 3H QTLs share homology. The 6HS QTL is identical to or closely linked to the SSLB resistance locus Rsp4 and the 1H QTL to the Rsp2 or Rsp3 locus. The 3H and 2HS QTLs are unique and offer new opportunities for pyramiding resistance genes through marker-assisted breeding for resistance to S. passerinii.     

Characterisation of target-site resistance to ACCase-inhibiting herbicides in the weed Alopecurus myosuroides (black-grass)

Resistance to aryloxyphenoxypropionate (AOPP), cyclohexanedione (CHD) and phenylurea herbicides was determined in UK populations of Alopecurus myosuroides Huds. Two populations (Oxford AA1, Notts. A1) were highly resistant (Resistance indices 13-->1000) to the AOPP and CHD herbicides fenoxaprop, diclofop, fluazifop-P and sethoxydim, but only marginally resistant to the phenylurea, chlorotoluron. Analyses of acetyl coenzyme A carboxylase (ACCase) activity showed that an insensitive ACCase conferred resistance to all the AOPP/CHD herbicides investigated. Another population, Oxford S1, showed no resistance to sethoxydim at the population level, but contained a small proportion of plants (<10%) with an insensitive ACCase. Genetic studies on the Notts A1 and Oxford S1 populations demonstrated that target site resistance conferred by an insensitive ACCase is monogenic, nuclearly inherited with the resistant allele showing complete dominance. Investigations of the molecular basis of resistance in the Notts A1 population showed that sethoxydim resistance in A myosuroides was associated with the substitution of an isoleucine in susceptible with a leucine in resistant plants, which has also been found in three other resistant grass-weed species (Setaria viridis (L) Beauv, Avena fatua L, Lolium rigidum Gaud).     

浙江稻区千金子对氰氟草酯和噁唑酰草胺的抗药性及其分子机制研究

千金子是中国直播稻田的优势禾本科杂草之一,严重威胁水稻的产量和品质。为了进一步明确浙江地区水稻田千金子对芳氧苯氧丙酸类除草剂的抗性发生情况,本研究从浙江部分稻区共采集了11个千金子种群 (其中1个为敏感种群),通过整株植物测定法检测了各种群对氰氟草酯和噁唑酰草胺的敏感性。结果显示:共有8个种群对氰氟草酯产生了抗性 (抗性指数为2.1~79.1),9个种群对噁唑酰草胺产生了抗性 (抗性指数为2.0~31.0),其中对氰氟草酯的抗性问题更为显著。在此基础上,通过基因扩增和克隆,对敏感种群和抗性种群的乙酰辅酶A羧化酶 (Acetyl-CoA carboxylase,ACCase) 基因部分序列进行比对,结果在3个抗性种群中发现突变,其中1个种群为Ile-1781-Val突变,另外2个种群则为Trp-2027-Cys突变。该研究结果表明,目前浙江部分稻区千金子种群已对氰氟草酯和噁唑酰草胺产生了抗药性,其中靶标酶基因突变是导致部分种群产生抗药性的原因之一。     

小麦-滨麦易位系M8657-1抗条锈病基因遗传分析和分子标记

 M8657-1, one of the wheat translocation lines derived from Leymus mollis Trin. Hara, is possessed of effective resistance at all stages to Su-ll and other dominant races of Puccinia striiformis f. sp. tritici in China. Seedlings of the parents, F₁, and F₂ progeny derived from the cross of M8657-1 (resistant) Mingxian169 (susceptible) were inoculated with Su-ll in greenhouse to identify and map the probable new stripe rust resistance gene. The results suggested that the stripe rust resistance in M8657-1 was conferred by a pair of recessive genes. Simple sequence repeat (SSR) technique was used to detect molecular marker associated with the resistance gene:208 pairs of wheat SSR primers were used to screen the two parents, as well as resistant and susceptible bulks and then three SSR markers were selected for genotyping the F₂ population. The geue, temporarily designated as YrLml, was found to be located on the chromosome 7DL and flanked by three SSR markers GDM67, WMC150 and WMC671, with the genetic distance of 5.0, 9.7 and 11.8cM, respectively.     

野生稻与栽培稻的杂种后代对褐稻虱的抗性机制初探

为了开拓抗虫基因源,培育抗性稳定的水稻抗虫品种,采用感虫的栽培稻雄性不育株为母本,以高抗褐稻虱的野生稻为父本进行有性杂交。通过用改良的苗期群体筛选法,对其后代进行抗性鉴定,证实其抗性可以遗传,已成功地获得了抗褐稻虱的杂种后代。并根据飞虱在抗虫植株上的生存率、群体建立、蜜露量和稻株的受害级别等指标与感虫对照种TN1上的差异显著性作比较,综合评定杂种后代对褐稻虱的抗性机制。结果表明,测试的N482(A)/101392组合的F_5、F_6、F_7和H_4等几个后代对褐稻虱的抗性机制均为抗生性。     

中粱93447抗条锈病基因遗传分析和分子作图

 用小麦条锈菌生理小种对中梁93447与感病品种铭贤169的杂交后代F₁、F₂和F₃代进行苗期温室抗条锈性遗传分析,结果表明中梁93447对CYR30的抗病性由1对显性基因控制。用中梁93447×铭贤169 F₂代分离群体建立抗、感DNA池,在F₂代群体中通过SSR标记技术寻找与抗病基因连锁的分子标记,发现7个位于5BS上的标记Xwmc813、Xwmc740、Xgwm159、Xbarc4、Xwmc616、Xwmc363和Xbarc89与目的基因连锁,遗传距离分别为17、12.9、8.3、4.5、3.7、9.1和20.8cM。系谱分析及分子标记分析表明,该基因可能来自中间偃麦草,暂命名为YrZL93447。与已定位于5B染色体上的抗条锈病基因的比较研究表明,YrZL93447可能是1个新的抗条锈病基因。用SSR引物BARC4和WMC616检测43个黄淮麦区主栽品种,其中7%的黄淮麦区主栽品种具有与YrZL93447基因相同的标记位点。     

Molecular marker mapping of leaf rust resistance gene lr46 and its association with stripe rust resistance gene yr29 in wheat

ABSTRACT Leaf and stripe rusts, caused by Puccinia triticina and P. striiformis, respectively, are globally important fungal diseases of wheat that cause significant annual yield losses. A gene that confers slow rusting resistance to leaf rust, designated as Lr46, has recently been located on wheat chromosome 1B. The objectives of our study were to establish the precise genomic location of gene Lr46 using molecular approaches and to determine if there was an association of this locus with adult plant resistance to stripe rust. A population of 146 F(5) and F(6) lines produced from the cross of susceptible 'Avocet S' with resistant 'Pavon 76' was developed and classified for leaf rust and stripe rust severity for three seasons. Using patterns of segregation for the two diseases, we estimated that at least two genes with additive effects conferred resistance to leaf rust and three to four genes conferred resistance to stripe rust. Bulked segregant analysis and linkage mapping using amplified fragment length polymorphisms with the 'Avocet' x 'Pavon 76' population, F(3) progeny lines of a single chromosome recombinant line population from the cross 'Lalbahadur' x 'Lalbahadur (Pavon 1B)', and the International Triticeae Mapping Initiative population established the genomic location of Lr46 at the distal end of the long arm of wheat chromosome 1B. A gene that is closely linked to Lr46 and confers moderate levels of adult plant resistance to stripe rust is identified and designated as Yr29.