A new rice gall midge resistance gene in the breeding line CR57-MR1523, mapping with flanking markers and development of NILs

Gall midge is the third most destructive insect pests of rice after stem borers and planthoppers. Host plant resistance has been recognized as the most effective and economic, means for gall midge management. With the characterization of a new gall midge biotype (GMB) 4M, unique feature of gall midge resistance in the breeding line CR57-MR1523 was highlighted. Multi-location evaluation of F₃ families derived from the cross TN1 × CR57-MR1523 against different gall midge biotypes helped to identify a new dominant gene conferring resistance against GMB4. This gene has been designated as Gm11t. Though CR57-MR1523 has been extensively used in breeding gall midge resistant rice varieties like Suraksha, neither the genetics of resistance nor chromosomal location of the resistance gene(s) is known. In the present study we have tagged and mapped the new gall midge resistance gene, Gm11t, on chromosome 12, using SSR markers. To map the gene locus, 466 F₁₀ generation Recurrent Inbred Lines (RILs), from the cross of TN1 × CR57-MR1523 were used. Of the 471 SSR markers spread across the rice genome, 56 markers showed polymorphism and were used to screen a subset of the mapping population consisting of 10 resistant (R) and 10 susceptible (S) F₁₀ RILs. Six SSR markers, RM28706, RM235, RM17, RM28784, RM28574 and RM28564 on chromosome 12 were initially found to be associated with resistance and susceptibility. Based on the linkage analysis in selected 158 RILs, we were able to map the locus between two flanking SSR markers, RM28574 and RM28706, on chromosome 12 within 4.4 and 3.8 cM, respectively. Further, two NILs with 99% genetic similarity, were identified from the RILs which differed in gall midge resistance. The tightly linked flanking SSR markers will facilitate marker-assisted gene pyramiding and map-based cloning of the resistant gene. NILs would be valuable materials for functional analysis of the identified candidate gene.     

Flanking SSR markers for allelism test for the Asian rice gall midge (Orseolia oryzae) resistance genes

Screening of rice germplasm against Asian rice gall midge, Orseolia oryzae (Wood-Mason), biotypes in India has led to identification of over 300 resistant rice genotypes. However, only ten resistance genes have been characterized so far. Identification of new genes through classical allelism test is tedious and time consuming. We propose to use closely linked flanking Simple Sequence Repeat (SSR) markers in allelism tests for identification of resistance genes. Of the ten known gall midge resistance genes, eight have been tagged and mapped. The Gm1 and Gm2 genes have closely linked flanking markers. Hence SSR markers RM219 and RM444, flanking the gene Gm1, and RM317, RM241 along with the SCAR marker F8, flanking the gene Gm2, were selected for this study. Tests with one set of 13 genotypes likely to carry Gm1 and another set of 17 genotypes suspected to contain Gm2 suggested the presence of the respective allele in all the 13 and 15 genotypes from these sets, respectively. Classical allelism test perfectly matched with the markers test. There were two exceptions involving amplification with RM444 in cultivar Kavya and with RM241 in genotype AE20, suggesting a single recombination which could have resulted in the mismatch. All the three markers in the genotype Bhumansan and the two flanking markers RM317 and F8 in AE20 indicated the absence of the Gm2 allele. This was validated through a classical test, revealing a segregation ratio of 15 resistant: 1 susceptible F₂ progeny of both the crosses between the Gm2 source Phalguna and these genotypes. We performed the allelism test with the markers on another set of 56 randomly selected gall midge resistant genotypes to discover possible sources of new resistance genes.     

华南抗稻瘿蚊分子标记辅助育种

亚洲稻瘿蚊(Orseolia oryzae Wood-Mason)是华南的主要水稻害虫.选育抗虫品种是最有效的生态控制方法.本文综述1998-2006年抗性育种的进展.用AFLP方法对从中国广东省7个地点采集的4个生物型的DAN指纹进行分析;在对用RAPD和SSR技术分别对抗中国4个稻瘿蚊生物型的基因Gm6精细定位基础上,用与Gm6紧密连锁的STS和SSR标记开展分子标记辅助育种,创造了一批抗稻瘿蚊的新种质,包括育成了6个栽培稻和6个二系杂交稻和1个三系杂交稻并在农户试种,在中国广东成功地建立了分子标记辅助选育抗稻瘿蚊品种的技术体系.     

Analysis of rice blast resistance gene <Emphasis Type="Italic">Pi</Emphasis>-<Emphasis Type="Italic">z</Emphasis> in rice germplasm using pathogenicity assays and DNA markers

The Pi-z gene in rice confers resistance to a wide range of races of the rice blast fungus, Magnaporthe oryzae. The objective of this study was to characterize Pi-z in 111 rice germplasm accessions using DNA markers and pathogenicity assays. The existence of Pi-z in rice germplasm was detected by using four simple sequence repeat (SSR) markers (RM527, AP4791, AP5659-1, AP5659-5) closely linked to Pi-z, and was verified using pathogenicity assays with an avirulent strain (IE1k) and two virulent races (IB33 and IB49). Among 111 germplasm accessions evaluated, 73 were found to contain the Pi-z gene using both SSR markers and pathogenicity assays. The remaining 38 germplasm accessions were found to be inconsistent in their responses to the blast races IB33, IEIk and IB49 with expected SSR marker alleles, suggesting the presence of unexpected SSR alleles and additional R gene(s). These characterized germplasm can be used for genetic studies and marker-assisted breeding for improving blast resistance in rice.     

Genetic analysis and pyramiding of two gall midge resistance genes (Gm-2 and Gm-6t) in rice ( Oryza sativa L.)

Asian rice gall midge (Orseolia oryzae) is a major pest across much of south and southeast Asia. This pest is genetically diverse and many gall midge biotypes are known to exist in each country. During the last three decades, host plant resistance has proved to be the most effective mechanism of controlling the Asian rice gall midge. Seven genes conditioning resistance to gall midge larvae have been identified in rice (Oryza sativa) and are being used in cultivar improvement programs. However, some of these genes are rendered ineffective by new gall midge biotypes. Increased understanding of genetics, inheritance, allelic relationships and linkage is necessary to maximise the durability of major gene resistance by the pyramiding of these genes. The two genes, Gm-2 and Gm-6(t), are known to confer resistance against a number of biotypes in India and China, respectively. An F₃ population derived from a cross between Duokang #1 (donor of Gm-6(t)) and Phalguna (donor of Gm-2) was screened against Chinese gall midge biotype 4 at Guangdong, China, and Indian gall midge biotype 1 at Raipur, India. At each location, separately,a single gene governed resistance. The parallel segregation of 417 F₃progenies for both biotypes at two locations revealed that recombination had occurred between the two genes, establishing that the two genes are not allelic. However, the two genes Gm-2 and Gm-6(t), were found to be linked with a distance of ∼16.3 cM. A number of lines homozygous at one locus and segregating for the other locus were identified and selected. These lines were selfed to obtain lines homozygous for the favourable alleles at both loci (two locus pyramids). This is the first report on use of conventional host-pest interaction method for pyramiding two closely located Gm-resistance loci of dissimilar effects. The implications of deployment of these pyramids within and across country borders, with reference to the prevailing gall midge populations are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

A novel blast resistance locus in a rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivar,Chumroo, of Bhutan

The rice cultivar ‘Chumroo’ is commonly cultivated in the mid- and high-altitude areas of Bhutan. This cultivar has shown durable blast resistance in that area, without evidence of breakdown, for over 20 years. Chumroo was inoculated with 22 blast isolates selected from the race differential standard set of Japan. The cultivar showed resistance to all the isolates. To identify the resistance gene(s), Chumroo was crossed with a susceptible rice cultivar, Koshihikari. The F₁ plants of the cross showed resistance. Segregation analyses of 300 F₃ family lines fitted the segregation ratio of 1:2:1 and indicated that a single dominant gene controls the resistance to a blast isolate Ao 92-06-2 (race 337.1). The Chumroo resistance locus (termed Pi46(t)) was mapped between two SSR markers, RM6748 and RM5473, on the terminal region of the long arm of chromosome 4, using linkage analysis with SSR markers. The nearest marker, RM5473, was linked to the putative resistance locus at a map distance of 3.2 cM. At the chromosomal region, no true resistance genes were identified, whereas two field resistance genes were present. Therefore, we designated Pi46(t) as a novel blast resistance locus.     

Molecular mapping of the fertility-restoration gene Rf4 for WA-cytoplasmic male sterility in rice

The wild abortive cytoplasmic male sterility (CMS‐WA) system, an ideal type of sporophytic CMS in indica rice, is used for the large‐scale commercial production of hybrid rice. Searching for restorer genes is a good approach when phenotyping is very time‐consuming and requires the determination of spikelet sterility in testcross progeny. To establish more precisely the genetical and physical maps of the Rf4 gene, high‐resolution mapping of this locus was carried out using simple sequence repeat (SSR) markers and newly designed markers in a F₂ population. The genetic linkage analysis indicated that five SSR markers (RM6737, RM304, RM171, RM5841 and RM228) on the long arm of chromosome 10 were linked with the Rf4 gene. Rf4 was flanked by two SSR markers RM171 and RM6737 at distances of 3.2 and 1.6 cM, respectively. Also, within the region between Rf4 gene and RM171, a newly designed primer pair, AB443, produced two sterile‐specific markers, AB443‐400 and AB443‐500, 0.5 and 1.03 cM from the gene. The flanking markers identified give promise for their application in molecular marker‐assisted selection (MAS) and they are also suitable for starting chromosome walking to clone Rf4 gene in the near future.     

Identification of microsatellite markers linked to the blast resistance gene <Emphasis Type="Italic">Pi-1(t)</Emphasis> in rice

The present work was conducted to identify microsatellite markers linked to the rice blast resistance gene Pi-1(t) for a marker-assisted selection program. Twenty-four primer pairs corresponding to 19 microsatellite loci were selected from the Gramene database (www. gramene.org) considering their relative proximity to Pi-1(t) gene in the current rice genetic map. Progenitors and DNA bulks of resistant and susceptible families from F₃ segregating populations of a cross between the near-isogenic lines C101LAC (resistant) and C101A51 (susceptible) were used to identify polymorphic microsatellite markers associated to this gene through bulked segregant analysis. Putative molecular markers linked to the blast resistance gene Pi-1(t) were then used on the whole progeny for linkage analysis. Additionally, the diagnostic potential of the microsatellite markers associated to the resistance gene was also evaluated on 17 rice varieties planted in Latin America by amplification of the specific resistant alleles for the gene in each genotype. Comparing with greenhouse phenotypic evaluations for blast resistance, the usefulness of the highly linked microsatellite markers to identify resistant rice genotypes was evaluated. As expected, the phenotypic segregation in the F₃ generation agreed to the expected segregation ratio for a single gene model. Of the 24 microsatellite sequences tested, six resulted polymorphic and linked to the gene. Two markers (RM1233*I and RM224) mapped in the same position (0.0 cM) with the Pi-1(t) gene. Other three markers corresponding to the same genetic locus were located at 18.5 cM above the resistance gene, while another marker was positioned at 23.8 cM below the gene. Microsatellite analysis on elite rice varieties with different genetic background showed that all known sources of blast resistance included in this study carry the specific Pi-1(t) allele. Results are discussed considering the potential utility of the microsatellite markers found, for MAS in rice breeding programs aiming at developing rice varieties with durable blast resistance based on a combination of resistance genes. Centro Internactional de Agricultura Tropical (CIAT) institute where the research was carried out     

Development and validation of microsatellite markers linked to the rice blast resistance gene <Emphasis Type="Italic">Pi-z</Emphasis> of Fukunishiki and Zenith

Rice blast resistance gene ‘Pi-z’ present in rice genotypes, Zenith and Fukunishiki, represents a potential source of blast resistance for the north-western Himalayan region of India. We tested the reliability of microsatellite markers linked to Pi-z for assessing blast resistance phenotype in crosses of commercial importance. A new set of microsatellite markers linked to Pi-z was also developed by exploiting the publicly available marker and genomic resources of rice. Of the three previously reported markers for Pi-z, only MRG5836 was suitable for the marker assisted selection of Pi-z. Among the 17 microsatellites selected from the putative region of Pi-z locus, two, RM8225 and RM8226 cosegregated with MRG5836 and were located at distance of 1.2–4.5 cM from the gene. A new microsatellite marker ‘SSR236’ was developed from the (CT)₁₆ repeat of PAC clone P0502B12, which exhibited closer linkage (0.6–1.2 cM) to Pi-z. Survey of the allelic diversity at the loci of the Pi-z linked microsatellite markers revealed that the Fukunishiki and Zenith type alleles were not present in majority of the local indica rice genotypes. As these markers are polymorphic between the Pi-z donors and a great majority of local indica rices tested, they can be used as a selection tool in rice breeding programs aimed at improving the blast resistance of local rices.     

A new gene for resistance in rice to Asian rice gall midge (Orseolia oryzae Wood Mason) biotype 1 population at Raipur, India

The Asian rice gall midge, Orseolia oryzae Wood Mason (Diptera: Cecidomyiidae), is a major pest of rice in several South and South East Asian countries. The maggots feed internally on the growing tips of the tillers and transform them into tubular galls, onion leaf-like structures called ‘silver shoots’ resulting into severe yield loss to the rice crop. We studied the mode of inheritance and allelic relationships of the resistance genes involved in resistant donor Line 9, a sib of a susceptible cultivar ‘Madhuri’. The segregation behaviour of F₁, F₂ and F₃ populations of the cross between Line 9 and susceptible cultivar MW10 confirmed the presence of a single dominant gene for resistance. Tests of allelism with all the known genes giving resistance to this population indicated that Line 9 possessed a new gene which was designated Gm 9 This revised version was published online in July 2006 with corrections to the Cover Date.     

Validation of molecular markers linked to fertility restorer gene(s) for WA-CMS lines of rice

The present study was carried out with the objective to validate the molecular markers, which have been previously reported to be linked to fertility restorer (Rf) gene(s) for WA-CMS lines of rice. Two mapping populations involving fertility restorer lines for WA-cytoplasm, viz., (i) an F₂ population derived from the cross IR58025A/KMR3R consisting of 347 plants and (ii) a BC₁F₁ population derived from the cross IR62829A/IR10198R//IR62829A consisting of 130 plants were analyzed. Nine SSR and three CAPS markers reported to be linked to Rf genes along with two previously unreported SSR markers were analyzed in the mapping populations. In both the populations studied, the trait of fertility restoration was observed to be under digenic control. Eight SSR markers (RM6100, RM228, RM171, RM216, RM474, RM311, MRG4456 and pRf1&2) showed polymorphism between the parents of the F₂ population, while the SSR markers RM6100 and RM474 showed polymorphism between the parents of both the F₂ and BC₁F₁ populations. Only one CAPS marker, RG146FL/RL was polymorphic between the parents of the BC₁F₁ population. RM6100 was observed to be closely segregating with fertility restoration in both the mapping populations and was located at a distance of ~1.2 cM. The largest phenotypic variation was accounted for the region located between RM311 and RM6100. Using the marker-trait segregation data derived from analysis of both the mapping populations, a local linkage map of the genomic region around Rf-4, a major fertility restoration locus on Chromosome 10 was constructed, and RM6100 was observed to be very close to the gene at a distance of 1.2 cM. The accuracy of the marker RM6100 in predicting fertility restoration was validated in 21 restorers and 18 maintainers. RM6100 amplified the Rf-4 linked allele in a majority of the restorers with a selection accuracy of 94.87%. Through the present study, we have established the usefulness of the marker RM6100 in marker-assisted selection for fertility restoration in segregating populations and identification of restorers while screening rice germplasm for their fertility restoration ability.     

Detection of quantitative trait locus for leaffolder (Cnaphalocrocis medinalis (Guenée)) resistance in rice on linkage group 1 based on damage score and flag leaf width

Rice leaffolder (RLF) (Cnaphalocrocis medinalis (Guenée) is a destructive and widespread insect pest throughout the rice growing regions in Asia. The genetics of resistance to RLF in rice is very complex and not thoroughly explored. The present study was conducted to detect the quantitative trait loci (QTL) associated with RLF resistance involving 176 recombinant inbred lines (RILs) of F₈ generation derived from a cross between IR36, a leaffolder susceptible variety and TNAULFR831311, a moderately resistant indica rice culture. Simple sequence repeat (SSR) markers were used to construct specific linkage groups of rice. All the RILs were screened to assess their level of resistance to RLF by measuring the leaf area damaged. Besides this, the length and width of the flag leaf of each RIL were measured since these two parameters were considered as correlated traits to the RLF resistance in rice. All the above parameters observed across the RILs showed quantitative variation. Correlation analysis revealed that damage score based on greenhouse screening was positively correlated with length and width of the flag leaf. Out of 364 SSR markers analysed, 90 were polymorphic between the parents. Multi-point analysis carried out on segregating 69 SSR marker loci linkage group wise resulted in construction of linkage map with eleven groups of 42 SSR markers. Through single marker analysis, 19 SSR markers were found to have putative association with the three phenotypic traits studied. Of these markers, RM472 was identified as a locus having major effect on RLF resistance trait based on length of the flag leaf. Interval mapping detected two QTLs on linkage group 1. Among these QTLs, the QTL flanked by RM576–RM3412 were found to be associated with width of the flag leaf and RLF resistance. The putative SSR markers associated with leaffolder resistance identified in the present study may be one of the loci contributing resistance to RLF in rice.     

Chromosomal location of powdery mildew resistance gene Pm16 in wheat using SSR marker analysis

The use of resistant cultivars is a most economical way to control powdery mildew (Blumeria graminis f.sp. tritici) in wheat (Triticum aestivum L.). Identification of molecular markers closely linked to resistance genes can greatly increase the efficiency of pyramiding resistance genes in wheat cultivars. The objective of this study was to identify molecular markers closely linked lo the powdery mildew resistance gene Pm16. An F₂ population with 156 progeny was produced from the cross‘Chancellor’(susceptible) ב70281’ (resistant), A total of 45 SSR markers on chromosomes 4A and 5B of wheat and 15 SSRs on chromosome 3 of rice was used lo lest the parents, as well as the resistant and susceptible bulks: the resulting polymorphic markers were used to genotype the F₂ progeny. Results indicated that the SSR marker Xgwm159, located on the short arm of chromosome 5B, is closely linked to Pm16 (genetic distance: 5.3 CM). The cytogenetical data presented in an original report, in combination with this molecular analysis, suggests that Pm16 may he located on a translocated 4A.5BS chromosome.     

Genetic mapping and inheritance of Russian wheat aphid resistance gene in accession IG 100695

The Russian wheat aphid (RWA), Diuraphis noxia (Kurdjumov), is an important pest of small‐grain cereals, particularly wheat, worldwide. The most efficient strategy against the RWA is to identify sources of resistance and to introduce them into susceptible wheat genotypes. This study was conducted to determine the mode of inheritance of the RWA resistance found in ICARDA accession IG 100695, to identify wheat microsatellite markers closely linked to the gene and to map the chromosomal location of the gene. Simple sequence repeat (SSR) marker scores were identified in a mapping population of 190 F₂ individuals and compared, while phenotypic screening for resistance was performed in F_2 : 3 families derived from a cross between ‘Basribey’ (susceptible) and IG 100695 (resistant). Phenotypic segregation of leaf chlorosis and rolling displayed the effect of a single dominant gene, temporarily denoted Dn100695, in IG 100695. Dn100695 was mapped on the short arm of chromosome 7D with four linked SSR markers, Xgwm44, Xcfd14, Xcfd46 and Xbarc126. Dn100695 and linked SSR markers may be useful for improving resistance for RWA in wheat breeding.     

Genetic Analysis of Resistance to Gall Midge (Orseolia oryzae Wood Mason) in Rice

The inheritance of resistance to rice gall midge (Ranchi biotype) was studied in 12 resistant cultivars by crossing with susceptible cultivars. By the study of F₁, F₂, F₃, B₁ and B₂ generations, it was found that resistance was governed by a single dominant gene in ‘Surekha’, ‘Phalguna’, ‘Rajendra Dhan 202’, ‘IET 7918’‘IET 6187’, ‘BG 404-1’; by duplicate dominant genes in ‘W 1263’, ‘RPW 6-17’ and ‘WGL 48684’ and a monogenic recessive gene in ‘OB 677’ and ‘BKNBR 1008-21’. The allelism test of the resistant genes in the test cultivars with already known genes Gm1 and Gm2 was carried out. A single dominant gene that conveyed the resistance in ‘RPW 6–17’, ‘IET 7918’ and ‘IET 6187’ was allelic to Gm1 and segregated independently of Gm2. The resistance in ‘Phalguna’, ‘Rajendra Dhan 202’, ‘W 1263’ and ‘RPW 6–17’, ‘IR 36’ and ‘WGL 48684’ was governed by Gm2 gene which was independent of Gm1. Two additional genes were identified and designated as Gm3 and gm4. Three test cultivars ‘BG 404-1’, ‘W 1263’ and ‘WGL 48684’ were found to have Gm3 gene for resistance which was non-allelic and segregated independently of Gm1 and Gm2. Thus the cultivars ‘W 1263’ and ‘WGL 48684’ had two resistance genes Gw2 and Gm3 together. The cultivar ‘RPW 6–17’ also had two resistance genes Gm1 and Gm2 together. The recessive gene gm4 which conditioned the resistance in ‘OB 677’ and ‘BKNBR 1008-21’ was nonallelic to and segregated independently of Gm1, Gm2 and Gm3 genes. Linkage studies of the resistance gene with pigment characters were carried out in ‘Purple gora/IR 36’ cross. The resistance gene Gm2 was found to be linked with the genes governing the pigmentation in node, apiculus and stigma with crossover values of 15.78, 31.57 and 35.78 % respectively. By the trisomic analysis, it was found that the Gm2 gene was located on chromosome 3.     

Molecular mapping of a novel blast resistance gene Pi38 in rice using SSLP and AFLP markers

Blast, caused by Magnaporthe grisea, is the most devastating disease of rice worldwide. In this study, the main objective was to identify and map a new gene for blast resistance, in an indica rice cultivar ‘Tadukan’ against blast fungal isolate B157, using molecular tools. F₂ segregating population was derived from ‘CO39’ (susceptible) and ‘Tadukan’ (resistant), and molecular mapping of the blast resistance gene was carried out using simple sequence length polymorphism (SSLP) and amplified fragment length polymorphism (AFLP) methods. Two SSLP markers, RM206 and RM21 and three AFLP markers (AF1: E‐aca/M‐ctt; AF2: E‐aca/M‐cat and AF3: E‐acc/M‐cac2) were identified to be linked to the resistance gene. The co‐segregation analysis using SSLP markers implied that the blast resistance gene designated Pi38 resides on rice chromosome 11.     

Identification of QTLs for the resistance to rice stripe virus in the <Emphasis Type="Italic">indica</Emphasis> rice variety Dular

The indica variety Dular has a high level of resistance to rice stripe virus (RSV). We performed quantitative trait locus (QTL) analysis for RSV resistance using 226 F₂ clonal lines at the seedling stage derived from a cross between the susceptible japonica variety Balilla and the resistant indica variety Dular with two evaluation criteria, infection rate (IR) and disease rating index (DRI). The experiments were performed in both 2004 and 2005. Based on IR, three putative QTLs were detected and had consistent locations in the 2 years, one QTL was detected in the RM7324–RM3586 interval on chromosome 3. The other two QTLs were linked and located in the RM287–RM209 and RM209–RM21 intervals on the long arm of chromosome 11, and accounted for 87.8–57.8% of the total phenotypic variation in both years. Based on DRI, three putative QTLs were also detected and had consistent locations in both years. One of them was located in the RM1124–SSR20 interval on the short arm of chromosome 11, while the other two linked QTLs had the same chromosomal locations on chromosome 11 as those detected by IR, and accounted for 55.7–42.9% of total phenotypic variation in both years. In comparison to the mapping results from previous studies, one of the two linked QTLs had a chromosomal location that was similar to Stv-b ⁱ , an important RSV resistance gene, while the other appeared to be a newly reported one.     

Molecular tagging of a stripe rust resistance gene in <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most important diseases of common wheat (Triticum aestivum L.). China has the largest stripe rust epidemic areas in the world and yield losses can be large. Aegilops tauschii Coss, the D-genome progenitor of common wheat, includes two subspecies, tauschii and strangulata (Eig) Tzvel. The ssp. strangulata accession AS2388 is highly resistant to the prevailing physiological races of PST in China, and possesses a single dominant gene for stripe rust resistance. In order to tag this gene, AS2388 was crossed with the highly susceptible ssp. tauschii accession AS87. The parents, F₂ plants, and F_2:3 families were tested at adult plant stage in field trials with six currently prevailing races. Simple sequence repeat (SSR) primers were used to identify molecular markers linked to the resistance gene. SSR markers Xwmc285 and Xwmc617 were linked to the resistance gene on chromosome arm 4DS flanking it at 1.7 and 34.6 cM, respectively. Based on the chromosomal location, this gene temporarily designated as YrAS2388 is probably novel. The resistance in Ae. tauschii AS2388 was partially expressed in two newly developed synthetic hexaploid backgrounds.     

SSR对黑龙江省主栽水稻品种抗瘟基因Pi-1的检测分析

应用水稻稻瘟病抗性基因Pi-1紧密连锁的3个微卫星标记RM144、RM224和MRG4766对黑龙江省49主栽水稻品种(系)进行抗瘟基因Pi-1检测分析。结果表明,用3个与抗瘟基因Pi-1紧密连锁的SSR标记同时对抗瘟基因Pi-1有检测是非常有效的途径,检测到富士光等14个水稻品种(系)含有Pi-1抗瘟基因,明确了该基因在黑龙江省水稻品种(系)中的分布情况,为分子育种、合理搭配种植品种等提供了理论依据。     

Tagging of four fertility restorer loci for wild abortive—cytoplasmic male sterility system in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) using microsatellite markers

Tagging of restorer genes for wild abortive (WA) CMS source by studying a 222 individual plants from a F₂ population of a cross between IR58025A × IR42686R. The restorer line IR42686R that was used in this study had been previously derived through random mating composite population (RMCP) involving 12 parents facilitated by IR36 genetic male sterility. Four Rf genes were tagged to simple sequence repeats (SSR) markers on chromosomes 1, 7, 10, 12 by recessive class analysis. The recombination frequency between a positive marker and Rf locus was calculated using maximum likelihood estimator assuming that all the 46 extremely sterile individual plants were homozygous at the targeted Rf locus. The recombination frequency between the marker and the restorer trait were converted to genetic distances using Kosambi function. A new Rf locus designated as Rf7 on chromosome 12 was found to be linked to RM7003 at a genetic distance of 13.3 cM (LOD 6.12). We report here first, a new molecular marker (RM 6344) linked to Rf4 locus on chromosome 7 that was previously mapped by trisomic analysis. RM443 and RM315 were flanking the Rf3 gene at a genetic distance of 4.4 (LOD 10.29) and 20.7 cM (LOD 3.98) on chromosome 1, respectively. The Rf6 was flanked on both side with SSR markers RM258 and RM591 at a genetic distance of 4.4 (LOD 10.29) and 23.3 cM (LOD 3.39) located on chromosome 10. The random mating composite population is an excellent breeding approach to develop superior restorer lines and for pyramiding different Rf genes of different CMS systems. Rf genes tagged with closely linked SSR markers would be facilitating marker assisted selection (MAS) in hybrid rice breeding program by reducing time and workload for identifying potential restorers. L. Bazrkar and A. J. Ali equally contributed to this work.