Pyramiding of two bacterial blight resistance and a semidwarfing gene in Type 3 Basmati using marker-assisted selection

A traditional Type 3 Basmati rice cultivar grown in India is tall and lodges even under low nitrogen fertilizer dose. In addition to lodging, it is highly susceptible to several diseases and pests including bacterial blight (BB). BB resistance genes (Xa21 and xa13) and a semidwarfing gene (sd-1) were pyramided in Type 3 Basmati from a rice cultivar PR106-P2 using marker-assisted selection (MAS). Foreground selection for BB resistance genes, Xa21 and xa13 and reduced plant height gene, sd-1 was carried on the basis of linked molecular markers pTA248, RG136 and ‘h’, respectively. The BC₂F₃ progenies with both the BB resistance genes were highly resistant with lower lesion length than either of the genes individually. Background profiling of the selected 16 BC₂F₃ progenies was done using 95 anchored SSR and 12 ISSR markers. Among the selected 16 BC₂F₃ progenies, 38-5-2 and 38-5-36 closely clustered along with the recipient parent Type 3 Basmati showing above 85% genetic similarity with the same. Further selection was continued till F₅ generation for higher recovery for Type 3 Basmati characteristics. The desirable alleles of intermediate amylose content (wx) and aroma (fgr) loci of Type 3 Basmati were also tracked using the linked SSR markers. The BC₂F₅ pyramid lines T3-4, T3-5, T3-6 and T3-7 homozygous for the three target genes Xa21, xa13 and sd-1 from the donor parent with wx and fgr alleles of Type 3 Basmati had excellent cooking quality and strong aroma.     

Integrating marker assisted background analysis with foreground selection for identification of superior bacterial blight resistant recombinants in Basmati rice

Basmati rice is highly susceptible to bacterial blight (BB) caused by Xanthomonas oryzae pv. oryzae. Transfer of BB resistance genes from non‐Basmati sources to Basmati through cross‐hybridization requires strict monitoring for recovery of the desirable Basmati quality traits in the recombinants, which show complex inheritance pattern. We integrated background analysis using mapped microsatellite markers with foreground selection to identify superior lines that combine useful genes from a non‐Basmati BB resistance donor line IRBB55 with grain and cooking quality characteristics of the popular Basmati rice variety ‘Pusa Basmati 1’ (PB 1) employing backcross pedigree strategy. Foreground selection using linked markers ensured presence of two genes, xa13 and Xa21 for BB resistance from IRBB55, and the recurrent parent PB 1 allele for the waxy locus giving intermediate amylose content and maintainer allele at fertility restorer locus in the BC₁F₅ recombinants. Background analysis enabled selection of recombinants with recurrent parent genome to the extent of 86.3% along with the quality traits. The extent of introgression of non‐Basmati donor chromosome segments in the superior selections was estimated to be < 7.8 Mb and < 6.7 Mb in the xa13 and Xa21 linked genomic regions, respectively. Association mapping identified three quantitative trait loci, one each for 1000‐grain weight, fertile grains/panicle and cooked kernel length. The backcross‐pedigree breeding strategy facilitated recovery of additional desirable characteristics from the donor in some of the selections. The elite selection Pusa 1460‐01‐32‐6‐7‐67 with maximum genomic background and quality characteristics of the recurrent Basmati parent gave resistance reaction against BB, similar to that of the non‐Basmati resistant check variety and recorded an yield advantage of 11.9% over the best check in the multiplication agronomic trial in the Basmati growing region of India. This line, which has been released as a new variety in the name of ‘Improved Pusa Basmati 1’ for commercial cultivation in India, is an example of successful application of marker assisted selection to variety development.     

Multiplex PCR genotyping for five bacterial blight resistance genes applied to marker‐assisted selection in rice (Oryza sativa)

Bacterial blight (BB) is the most economically damaging disease of rice in Asia and other parts of the world. In this study, a multiplex PCR genotyping method was developed to simultaneously identify genotypes of five BB resistance genes, Xa4, xa5, Xa7, xa13 and Xa21. The resistance R alleles were amplified using five functional markers (FMs) to generate amplicons of 217, 103, 179, 381 and 595 bp in IRBB66. Amplicons of 198, 107, 87, 391 and 467 bp corresponded to susceptible alleles in Taiwanese japonica rice cultivars. In backcross breeding programmes, the multiplex PCR assay was integrated into selection from a population using BB resistance donor IRBB66 crossed to rice cultivar ‘Tainung82’. Two plants with homozygosity for Xa4, xa5, Xa7, xa13 and Xa21 were selected from 1100 BC₂F₂ plants. In addition, the five BB resistance genes were also accurately identified in F₂ populations. This multiplex PCR method provides a rapid and efficient method for detecting various BB resistance genes, which will assist in pyramiding genes to improve durability of BB resistance in Taiwanese elite rice cultivars.     

Marker‐assisted introgression of bacterial blight and blast resistance into IR 58025B,an elite maintainer line of rice

IR 58025A is a very popular wild‐abortive cytoplasmic male sterile (WA‐CMS) line of rice and is extensively used for hybrid rice breeding. However, IR 58025A and many hybrids derived from it possess mild aroma (undesirable in some parts of India) and are highly susceptible to bacterial blight (BB) and blast diseases. To improve IR 58025A for BB and blast resistance, we have introgressed a major dominant gene conferring resistance against BB (i.e. Xa21) and blast (i.e. Pi54) into IR 58025B, the maintainer line of IR 58025A. An introgression line of Samba Mahsuri (i.e. SM2154) possessing Xa21 and Pi54 genes in homozygous condition and fine‐grain type was used as donor parent, and backcross breeding strategy was adopted for targeted introgression of the resistance genes. PCR‐based molecular markers tightly linked to Xa21 and Pi54 were used for selection of BB‐ and blast‐resistant lines, while closely linked markers were used for identification of backcross‐derived plants devoid of Rf4 and aroma. At BC₂F₅, four backcross‐derived lines possessing resistance against BB and blast, devoid of aroma, high yield, short plant stature, long‐slender grain type and with recurrent parent genome recovery ranging from 88.8% to 98.6% were selected and advanced for further evaluation. The improved versions of IR 58025B, viz. SB54‐11‐143‐9‐44‐5, SB54‐11‐143‐9‐44‐98, SB54‐11‐143‐9‐44‐111 and SB54‐11‐143‐9‐44‐171, behaved as perfect maintainers when test‐crossed with WA‐CMS lines. Agronomically superior lines of improved IR 58025B are being converted to CMS line through backcrossing for developing high‐yielding and biotic stress‐resistant rice hybrids.     

The host background of rice influences the resistance expression of a three genes pyramid (xa5 + xa13 + Xa21) to bacterial blight (Xanthomonas oryzae pv. oryzae) pathotypes of Indian mainland and Bay islands

Bacterial blight (BB) is a major disease of rice for which host resistance is the only effective solution. The three genes pyramid xa5 + xa13 + Xa21 is recently the most utilized combination for developing resistant varieties through marker‐assisted breeding. Our study was carried out to elicit the detailed response of twenty lines possessing these three genes in five genetic backgrounds to twelve diverse BB pathotypes in India. The lines developed from ADT 47 variety showed incomplete resistance to most of the pathotypes, whereas susceptibility varied from 8.3% to 16.6% in ADT 43 and IR24, respectively. However, in IMP ASD16/60 and Improved Samba Mahsuri, complete resistance against all pathotypes was observed. The overall results confirmed that genetic background plays crucial role for the effective expression of xa5 + xa13 + Xa21 combination. Molecular studies did not reveal correlation between origin of pathotypes and their virulence potential. It is suggested to deploy Improved Samba Mahsuri, IMP ASD 16/60 and AD1306 varieties in the bacterial blight prone areas or use them as donors for realizing wider and durable resistance.     

Making an Indian traditional rice variety Mahsuri, bacterial blight resistant using marker-assisted selection

Mahsuri a popular traditional variety and the first rain-fed mega variety of the Indian sub- continent. It is highly susceptible to bacterial leaf blight caused by Xanthomonas oryzae pv.oryzae. Nine best performing families of Mahsuri pyramid containing four bacterial blight resistance genes (Xa4, xa5, xa13 and Xa21) were evaluated for agronomic, yield and its related characters viz. days to 50% flowering, plant height, number of tillers, panicle length, filled grains per panicle, grain weight, and yield under natural and disease pressure conditions for three consecutive wet seasons. In addition these pyramids were also evaluated for three different spacings to find out the optimum spacing under disease free and disease pressure conditions. Results revealed that under disease free conditions there was no significant difference between the pyramids and the parent for the characters evaluated in each spacing. However characters plant height, number of tillers, panicle length, filled grains per panicle, yield per plant and yield per sq. m. showed significant variation between the different spacings across seasons. Under heavy disease pressure the parent exhibited highly susceptible reaction whereas the pyramid families were highly resistant. A wider spacing had less yield loss when compared to dense planting under BB infestation in case of parent. There was no such yield loss in the pyramid families. When yield per sq. m. was taken into consideration the 20 × 20 cm spacing showed the highest yield when compared to the other two spacings since number of plants were more. The pyramids insulated the yield loss against bacterial leaf blight and are a gain to the farmers to help overcome the heavy yield losses due to this disease. These pyramids have the potential to replace the parent and can be used directly. In addition they can be used as donors for bacterial blight resistance in any breeding program.     

Pyramiding of four resistance genes of bacterial blight in Tapaswini, an elite rice cultivar, through marker-assisted selection

Breeding for resistance against Bacterial blight (BB), a destructive disease in rice, is a major challenge because of the rich diversity of the pathogen, Xanthomonas oryzae pv. oryzae. Three resistance genes i.e. xa5, xa13 and Xa21 were transferred from IRBB 60 through marker-assisted backcross breeding to supplement the Xa4 gene present in Tapaswini, an elite cultivar having a wide coverage. The precise transfer was aided through effective foreground selection using STS markers RG 556, RG 136 and pTA248 linked to the three target genes, respectively. In the screening assays, the gene pyramids having four genes exhibited higher level of resistance against the disease. Background selection based on morphological and grain quality traits and SSR markers led to the recovery of 85–96 % of the recurrent parent genome in the gene pyramids. Multi location evaluation conducted over a two year period at several hotspot locations of bacterial blight in India has resulted in identification and release of CRMAS 2622-7-6 (IT-6), a promising gene pyramid as improved Tapaswini. The high levels of resistance exhibited in the improved genotype without any yield penalty and its similarity to the recurrent parent provides a successful demonstration of the potential of MAS in pyramiding genes as well as the recovery of the parental genome with high accuracy.     

Molecular marker-assisted selection for development of common bean lines resistant to angular leaf spot

The objective of this work was to develop homozygous common bean lines carrying angular leaf spot resistance genes derived from the cultivars ‘Mexico 54’, ‘MAR 2’ and ‘BAT 332’ through marker‐assisted selection. Molecular markers SCAR OPN02₈₉₀, RAPD OPE04₅₀₀ and OPAO12₉₅₀ linked to the resistance genes of ‘Mexico 54’, ‘MAR 2’ and ‘BAT 332’, respectively, were used in segregating backcross‐derived populations to selection. DNA fingerprinting was used to select homozygous BC₂F₃ and BC₁F₃ resistant plants genetically closer to the recurrent parent. Two homozygous BC₂F_2:3 and two and five BC₁F_2:3 families derived from ‘Ruda’ vs. ‘Mexico 54’ (RM), ‘MAR 2’ (RMA) and ‘BAT 332’ (RB) crosses were selected, respectively. After only one (RMA, RB) or two backcrosses (RM), five and eight BC₁F₃ lines derived from RMA and RB, respectively, and seven BC₂F₃ lines derived from RM, with 14.9–16.6, 16.9–18.6 and 9.3–11.1% of relative genetic distances to the recurrent parent were selected. This is the first report of lines resistant to angular leaf spot carrying genes of the cultivars ‘Mexico 54’, ‘MAR 2’ and ‘BAT 332’ developed with the aid of molecular markers.     

Use of molecular markers to accelerate the breeding of common bean lines resistant to rust and anthracnose

The main goal of this work was to introduce resistance genes for rust, caused by Uromyces appendiculatus, and anthracnose, caused by Colletotrichum lindemuthianum, in an adapted common bean cultivar through marker-assisted backcrossing. DNA fingerprinting was used to select plants genetically closer to the recurrent parent which were also resistant to rust and to race 89 of C. lindemuthianum. DNA samples extracted from the resistant parent (cv. Ouro Negro), the recurrent parent (cv. Rudá), and from BC₁, BC₂ and BC₃ resistant plants were amplified by the RAPD technique. The relative genetic distances in relation to the recurrent parent varied between 9 and 59% for BC₁, 7 and 33% for BC₂, and 0 and 7% for BC₃ resistant plants. After only three backcrosses, five lines resistant to rust and anthracnose with, approximately, 0% genetic distance in relation to the recurrent parent were obtained. These lines underwent field yield tests in two consecutive growing seasons and three of them presented a good yield performance, surpassing in that sense their parents and most of the reference cultivars tested.     

Pyramiding of Xa7 and Xa21 for the improvement of disease resistance to bacterial blight in hybrid rice

‘Minghui 63’ is a restorer line widely used in hybrid rice production in China for the last two decades. This line and its derived hybrids, including ‘Shanyou 63’, are susceptible to bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo). To improve the bacterial blight resistance of hybrid rice, two resistance genes Xa21 and Xa7, have been introgressed into ‘Minghui 63’ by marker‐assisted selection and conventional backcrossing, respectively. The single resistance gene‐introgressed lines, Minghui 63 (Xa21) and Minghui 63 (Xa7) had higher levels of resistance to bacterial blight than their derived hybrids, Shanyou 63 (Xa21) or Shanyou 63 (Xa7). Both Xa21 and Xa7 showed incomplete dominance in the heterozygous background of rice hybrids by infection with GX325 and KS‐1‐21. The improved restorer lines, with the homozygous genotypes, Xa21Xa21 or Xa7Xa7, were more resistant than their hybrids with the heterozygous genotypes Xa21xa21 or Xa7xa7. To further enhance the bacterial blight resistance of ‘Minghui 63’ and its hybrids, Xa21 and Xa7 were pyramided into the same background using molecular marker‐aided selection. The restorer lines developed with the resistance genes Xa21 and Xa7, and their derived hybrids were evaluated for resistance after inoculation with 10 isolates of pathogens from China, Japan and the Philippines, and showed a higher level of resistance to BB than the restorer lines and derived hybrids having only one of the resistance genes. The pyramided double resistance lines and their derived hybrids have the same high level of resistance to BB. These results clearly indicate that pyramiding of dominant genes is a useful approach for improving BB resistance in hybrid rice.     

Improving bacterial blight resistance of'6078', an elite restorer line of hybrid rice, by molecular marker-assisted selection

Bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (X00), is one of the most devastating diseases of rice world‐wide; it is also a serious problem of hybrid rice production in China. In this study, a molecular marker‐assisted introgression of Xa21, a gene highly resistant to a broad spectrum of Xoo strains, from ‘IRBB21’ was performed to improve the BB resistance of‘6078′, a new restorer line with high yielding potential. The entire process took one generation of crossing followed by three generations of backcrossing and one generation of selfing. The presence of Xa21 in each generation was determined by both polymerase chain reaction (PCR) and pathogen inoculation. Recombinations between Xa21 and flanking markers were identified by PCR analysis. Background selection was conducted in BC₁F₁ and BC₂F₁ using amplified fragment length polymorphism (AFLP) markers detecting a total of 129 polymorphic bands between‘6078’ and ‘IRBB21′. The individual selected in BC₃F₂, or‘6078′(Xa21), carried a fragment of less than 3.8 cM from the donor line in the Xa21 region on chromosome 11, and about 98.8% of the genetic background from the recurrent parent. The results showed that‘6078′(Xa21) had the same level and spectrum of BB resistance as the donor parent ‘IRBB21′, while maintaining the agronomic performance and combining ability of the original 6078. A significant increase in BB resistance was also achieved in the hybrid using 6078(Xa21) as the restorer line.     

Marker assisted backcross breeding approach to improve blast resistance in Indian rice (Oryza sativa) variety ADT43

Marker assisted backcrossing breeding has become one of the essential tools in transferring novel genes to adapted varieties and was employed to pyramid three blast resistance genes Pi1, Pi2 and Pi33 to a popular susceptible rice variety ADT43. Gene pyramiding process was facilitated by marker aided selection for both foreground as well as background genotype. Previously reported linked molecular markers were deployed to survey resistant and susceptible genotypes. In the BC₃F₁ generation four lines viz, AC-B3-11-7, AC-B3-11-36, AC-B3-11-57, AC-B3-11-83 were identified to be pyramided with three genes and subjected to background analysis and a genome recovery up to 95 % was observed and advanced to further generations. Morphological, yield and grain quality traits were significantly similar to ADT43. The introgressed lines with three gene combinations were highly resistant to the blast pathogen compared to genotypes with single genes and the susceptible checks under blast nursery screening at two epiphytotic locations; Coimbatore and Gudalur. The selected three gene pyramided backcross lines in the desirable background were advanced to obtain an improved ADT 43 with resistance to blast disease.     

Introgression of bacterial blight resistance genes Xa7, Xa21, Xa22 and Xa23 into hybrid rice restorer lines by molecular marker-assisted selection

Four bacterial blight (BB) resistance genes, Xa7, Xa21, Xa22 and Xa23, were introgressed into an elite hybrid rice restorer line Huahui 1035, by marker-assisted selection (MAS). Ten promising BB resistant lines identified by MAS approach in Huahui 1035 restorer background and their respective F₁ hybrids with a cytoplasmic male sterile line i.e. Jinke 1A, three BB resistant gene donors and their recurrent parent were evaluated for BB resistance by inoculating them with eleven representative races of Xanthomonas oryzae pv. oryzae (Xoo) in China. Further, the ten marker assisted selection of BB (MAS-BB) resistant restorer lines and their F₁ hybrids were also characterized for agro-morphological traits and grain yield. Results revealed that restorer lines with Xa23 introgression i.e. HBQ809 and HBQ810 were found to be resistant to all eleven Chinese representative Xoo races, showing broad spectrum resistance to BB. However, the lines possessing Xa7 or Xa7+Xa21 were resistant to ten out of eleven Xoo races. While restorer lines with Xa21 were resistant to nine out of eleven Xoo races. Interestingly, the F₁ hybrids with Xa23, Xa7 or Xa7+Xa21 were resistant to two severe epidemic Xoo races of China. Restorer lines i.e. HBQ807 and HBQ808 possessing Xa22 were only resistant to six or five out of inoculated eleven Xoo races. Lesion length comparisons between restorer lines and their F₁ hybrids for the above four BB resistance genes showed them to be dominant in heterozygote genotype. Three promising high yielding F₁ hybrids with BB resistance were identified for immediate exploitation for hybrid rice production in China.     

水稻抗白叶枯病基因的聚合育种

白叶枯病是严重危害水稻生产的重要病害之一,利用聚合多个抗性基因的水稻品种是控制该病害的有效途径。本研究以携带Xa4、xa5、xa13、Xa214个抗白叶枯病基因的IRBB60与8个水稻新品系,组配8个杂交组合.应用分子标记辅助选择技术从F2和F3群体中共获得216个携带4个抗白叶枯病基因的纯合体。用华南地区的5个白叶枯病病原菌小种进行田间接种的试验表明,4个抗性基因聚合系的抗性水平高于只带1个抗性基因的近等基因系。这些聚合系可用作华南地区高抗白叶枯病水稻品种育种的亲本材料。     

Development of B-Genome Chromosome Addition Lines of B. napus Using Different Interspecific Brassica Hybrids

By interspecific hybridization within the genus Brassica, trigenomic haploids were produced and back-crossed four times with B. napus, variety ‘Andor’. From this material, monosomic B-genome chromosome addition lines were selected with the extra chromosome derived from three different B-genome sources, i.e., B. nigra (BB), B. carinata (BBCC), and B. juncea (AABB). After selfing and/or microspore culture, disomic addition lines were obtained. Meiotic behavior was studied of the trigenomic hybrids, the pentaploid BC₁ plants, and the monosomic addition lines. The addition lines were shown to possess cytological stability and good fertility.     

Inheritance of resistance to Xanthomonas campestris pv. translucens in triticale

Summary Three triticale lines, Siskiyou, M2A-Beagle, and OK 77842 have been reported to possess resistance to bacterial leaf streak caused by Xanthomonas campestris, pv. translucens (Xct.). The three resistant lines were crossed to susceptible lines and crossed with each other. F₂, BC₁-F₁, BC₂-F₁ plants were inoculated with a mixture of two Xct strains. The segregation data indicate the presence of a single dominant gene in each of the three resistant lines to bacterial leaf streak. These three genes are either the same or closely linked herein designated as Xct₁.     

Expression levels of three bacterial blight resistance genes against K3a race of Korea by molecular and phenotype analysis in japonica rice (O. sativa L.)

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo) is a destructive disease of rice in the major rice growing countries of Asia. In 2003, a serious bacterial blight epidemic occurred in the southwestern coastal areas in Korea, causing significant yield loss due to the emergence of a new race, K3a. IR24 near-isogenic lines containing Xa4, xa5, Xa7 and Xa21 genes conferred different degrees of resistance to the most virulent K3a isolate, HB01009 in an inoculation experiment in the greenhouse. Expression levels of the resistance genes, Xa4, xa5 and Xa21 were studied in two F2 populations derived from the crosses between elite japonica cultivars and an advanced backcross breeding line possessing Xa4, xa5 and Xa21 genes. F₂ progenies segregated for K3a resistance (R) and susceptible (S) phenotypes in a ratio of 3(R):1(S) indicated that K3a resistance was controlled by a major dominant gene. Three PCR markers tightly linked to the resistance genes Xa4, xa5 and Xa21 confirmed the presence of the genes and their interaction with each gene. This study demonstrated that the Xa21 gene dominantly contributed to K3a resistance. However, the Xa4 gene also contributed to the full expression of resistance. The level of expression of strong resistance to K3a race was attributed to the presence of Xa21 and Xa4 genes irrespective of the presence of xa5 gene. Our results suggest that the R-gene combinations of Xa4+Xa21 could be a useful and effective strategy toward improving resistance to K3a race of Korean japonica cultivars.     

Rapid and high-precision marker assisted backcrossing to introgress the <Emphasis Type="Italic">SUB1</Emphasis> QTL into BR11, the rainfed lowland rice mega variety of Bangladesh

Flooding is one of the major hazards of rice production for the rainfed lowland rice ecosystem, and tolerant cultivars are urgently needed to help protect farmers from submergence damage. A quick and efficient strategy was implemented to introgress SUB1, a major QTL for submergence tolerance, into a rainfed lowland mega variety BR11 of Bangladesh by only two backcrosses and one selfing generation. In marker-assisted backcrossing (MABC), one tightly-linked simple sequence repeat (SSR) and two gene-based markers, four flanking SSR and 116 background SSR markers were used for foreground, recombinant and background selection, respectively, in backcrosses between a SUB1 donor IR40931-33-1-3-2 and BR11. BR11-Sub1, identified in a BC₂F₂ plant, possessed BR11 type SSR alleles on all fragments analyzed except the SUB1 QTL. The introgression size in BR11-Sub1 was 800 Kb indicating approximately 99.8% identity to BR11. BR11-Sub1 along with other introgression lines showed submergence tolerance similar to the tolerant parent. Yield, yield-component parameters and grain physico-chemical properties showed successful recovery of the BR11 traits in BR11-Sub1, with yield potential ranging from 5.2 to 5.6 t/ha, not significantly different from the recurrent parent mega variety BR11. Producing a large number (~1000) of backcross F₁ plants was considered essential to achieve recombination on both sides of the gene, limiting linkage drag with only two backcrosses. A large number of background markers ensured proper recovery of the recurrent parent genome in the BC₂F₂ generation. The study demonstrates a rapid and highly precise strategy to introgress a major QTL by BC₂F₂ generation into a modern rice variety using an unadapted donor. The variety can replace BR11 on more than 2 million of ha in Bangladesh and provide major increases in rice production.     

Mapping of a gene for leaf scald resistance in barley line 'B87/14' and validation of microsatellite and RFLP markers for marker-assisted selection

Seedlings of the barley line ‘B87/14’ were resistant to 22 out of 23 Australian isolates of Rhynchosporium secalis, the causal agent of leaf scald.‘B87/14’‐based populations were developed to determine the location of the resistance locus. Scald resistance segregated as a single dominant trait in BC₁F₂ and BC₁F₃ populations. Bulked segregant analysis identified amplified fragment length polymorphisms (AFLPs) with close linkage to the resistance locus. Fully mapped populations not segregating for scald resistance located these AFLP markers on chromosome 3H, possibly within the complex Rrs1 scald locus. Microsatellite and restriction fragment length polymorphism markers adjacent to the AFLP markers were identified and validated for their linkage to scald resistance in a second segregating population, with the closest marker 2.2 cM from the resistance locus. These markers can be used for selection of the Rrs.B87 scald‐resistance locus, and other genes at the chromosome 3H Rrs1 locus.     

AFLP-based STS markers closely linked to a fertility restoration locus (Rfm1) for cytoplasmic male sterility in barley

The Rfm1 gene restores the fertility of the msm1 and msm² male‐sterile cytoplasms in barley. Rfm1 is located on the short arm of chromosome 6H. To develop molecular markers tightly linked to Rfm1 for use in sophisticated marker‐assisted selection and map‐based cloning, an amplified fragment‐length polymorphism (AFLP) marker system with isogenic lines and a segregating BC₁F₁ population was used. Nine hundred primer combinations were screened and a linkage map was constructed around the Rfm1 locus by using 25 recombinant plants selected from 214 BC₁F₁ plants. Three AFLP markers were identified, e34m2, e46m19 and e48m17, linked to the locus. The most closely linked markers were e34m2, at 1.0 cM distally and e46m19, at 1.1 cM proximally. The two AFLP markers were converted to dominant STS markers. These markers should accelerate programmes for breeding restorer lines and will be useful for map‐based cloning.