Molecular markers for late blight resistance breeding of potato: an update

Late blight is the most devastating disease of the potato crop that can be effectively managed by growing resistant cultivars. Introgression of resistance (R) genes/quantitative trait loci (QTLs) from the Solanum germplasm into common potato is one of the plausible approaches to breed resistant cultivars. Although the conventional method of breeding will continue to play a primary role in potato improvement, molecular marker technology is becoming one of its integral components. To achieve rapid success, from the past to recent years, several R genes/QTLs that originated from wild/cultivated Solanum species were mapped on the potato genome and a few genes were cloned using molecular approaches. As a result, molecular markers closely linked to resistance genes or QTLs offer a quicker potato breeding option through marker‐assisted selection (MAS). However, limited progress has been achieved so far through MAS in potato breeding. In near future, new resistance genes/QTLs are expected to be discovered from wild Solanum gene pools and linked molecular markers would be available for MAS. This article presents an update on the development of molecular markers linked to late blight resistance genes or QTLs by utilization of Solanum species for MAS in potato.     

Linkage and quantitative trait locus mapping of foliage late blight resistance in the wild species Solanum vernei

The global cultivation of potato (Solanum tuberosum) is threatened by epidemics caused by new variants of the late blight pathogen, Phytophthora infestans. New sources of durable late blight resistance are urgently needed and these may be found in wild Solanum species. The diploid wild species, S. vernei, has not previously been subjected to mapping of quantitative trait loci (QTLs) for late blight resistance. Two populations designated HGIHJS and HGG, originating from a cross between a clone of S. vernei and two different S. tuberosum clones were evaluated in field trials for late blight infestation. The relative area under the disease progress curve (RAUDPC) was estimated and used for QTL mapping. A linkage map of S. vernei, comprising 11 linkage groups, nine of which could be assigned to chromosomes, was constructed. Results indicated that the resistance in S. vernei was quantitatively inherited. Significant QTLs for late blight resistance were identified on chromosomes VIII (HGG), VI and IX (HGIHJS). In addition, potential QTLs were detected on chromosomes VII (HGIHJS) and IX (HGG). A putative and a significant QTL for tuber yield were found on chromosomes VI and VII in HGG, but no linkage between yield and resistance was indicated. The QTL for late blight resistance, which mapped to chromosome IX, could be useful for late blight resistance breeding as it was located close to the microsatellite marker STM1051 in both populations.     

Differentiation of the high molecular weight glutenin subunit D t x 2.1 of Aegilops tauschii indicated by partial sequences of its encoding gene and SSR markers

The devastating late blight pathogen Phytophthora infestans infects foliage as well as tubers of potato. To date, resistance breeding has often focused on foliage blight resistance, but tuber blight resistance is becoming more and more important in cultivated potatoes. In this study, a reliable tuber assay for resistance assessment was developed and foliage and tuber blight resistance (R) was compared in four mapping populations. In the RH4X-103 population, tuber blight resistance inherited independently from foliage blight resistance. Three specific R genes against P. infestans were segregating. The Rpi-abpt and R3a genes function as foliage-specific R genes, whereas the R1 gene acts on both foliage and tuber. In the segregating populations SHRH and RH94-076, tuber and foliage blight resistance correlated significantly, which suggests that resistance in foliage and tuber is conferred by the same gene (could be R3b) and quantitative trait loci (QTL), respectively. In the CE population neither tuber nor foliage resistance was observed.     

Mapping polygenes for tuber resistance to late blight in a diploid Solanum phureja × S. stenotomum hybrid population

Potato tuber blight is a disease caused by the oomycete Phytophthora infestans (Mont.) de Bary. Due to the significant economic impact of this disease, introgression of durable resistance into the cultivated potato is one of the top priorities of breeding programmes worldwide. Though numerous resistance loci against this devastating disease have already been mapped, most of the detected loci are contributing towards foliar resistance while specific information on tuber resistance is limited. To identify the genetic components of tuber resistance and its relationship to foliar resistance and plant maturity we have investigated the host‐pathogen interaction in a segregating diploid hybrid Solanum phureja × S. stenotomum family. Mature tubers from this mapping family were inoculated with a sporangial suspension of P. infestans (US‐8 clonal lineage) and evaluated for lesion expansion. No significant correlation was detected between late blight resistance in foliage and tubers, and between plant maturity and tuber resistance. Four chromosomal regions were significantly associated with tuber resistance to the disease. The largest effect was detected near the marker locus PSC (LOD 10.7) located on chromosome 10. This locus explained about 63% of the total phenotypic variation of the trait. The other three resistance‐related loci were mapped on chromosomes 8 (GP1282, LOD 4.4), 6 (CP18, LOD 4.0) and 2 (CP157, LOD 3.8). None of the four tuber resistance loci coincides with the foliage resistance loci detected in this same family. Tuber blight resistance quantitative trait loci (QTL) on chromosomes 2, 8 and 10 are distinct from the maturity QTLs and have an additive effect on tuber resistance. These results indicate that different genes are involved in foliar and tuber resistance to P. infestans in the present family and that some of the resistance genes might be associated with late maturity.     

Mapping of rhizoctonia root rot resistance genes in sugar beet using pathogen response‐related sequences as molecular markers

Rhizoctonia root and crown rot caused by the fungus Rhizoctonia solani is a serious disease of sugar beet. An F_2:3 population from a cross between a resistant and a susceptible parent has been tested for R. solani resistance and a genetic map has been constructed from the corresponding F₂ parents. The map encompasses 38 expressed sequence tags (ESTs) with high similarity to genes which are involved in resistance reactions of plants (R‐ESTs) and 25 bacterial artificial chromosomes (BACs) containing nucleotide binding site (NBS)‐motifs typical for disease resistance genes. Three quantitative trait loci (QTL) for R. solani resistance were found on chromosomes 4, 5 and 7 collectively explaining 71% of the total phenotypic variation. A number of R‐ESTs were mapped in close distance to the R. solani resistance QTL. In contrast, the NBS‐BACs mapped to chromosomes 1, 3, 7 and 9 with two major clusters of NBS‐BACs on chromosome 3. No linkage between NBS‐BACs and R. solani resistance QTL was found. The data are discussed with regard to using R‐ESTs and NBS markers for mapping quantitative disease resistances.     

Genetic analysis and identification of two soybean mosaic virus resistance genes in soybean [Glycine max (L.) Merr]

Soybean mosaic virus (SMV) commonly affects soybean production worldwide, and the SC18 strain has been widespread in China. This study aimed to characterize and map the SC18 resistance genes present in soybean cultivars ‘Kefeng No. 1’ and ‘Qihuang 22’. Inheritance analysis revealed that two independent single dominant genes in Kefeng No. 1 and Qihuang 22 confer resistance to SC18. Using simple sequence repeat (SSR) markers and bulked segregant analysis, the Kefeng No. 1 and Qihuang 22 resistance genes were located on soybean chromosomes 2 and 13, respectively. We further screened two populations of recombinant inbred lines with 32 SSR markers in the target region, where the resistance gene in Kefeng No. 1 was fine mapped to an 80‐kb region containing six putative genes. Sequence and expression analyses of these genes revealed that SMV resistance in Kefeng No. 1 was probably attributable to three of the candidate genes (i.e. Glyma.02G127800, Glyma.02G128200 and Glyma.02G128300). Collectively, the results of this study will greatly facilitate the cloning of SC18 resistance genes and marker‐assisted breeding of SMV‐resistant soybean cultivars.     

Genetics of Resistance to Four Races of Xanthomonas campestris pv. oryzae in Some Rice Cultivars

Inheritance of resistance to four Philippine races of bacterial Might caused by Xanthomonas campestris pv. oryzae was investigated in four cultivars of rice, Oryza sativa L. Resistance to three races in ‘Benamuri’ and ‘Aus 192’ is governed by xa-5. In ‘Tepal Boro’ and ‘Bazail 975′, resistance to races 1, 2. and 3 is conferred by xa-5, but another recessive gene confers resistance to race 4. This recessive gene is closely linked to xa-5 and may be allelic to xa-13. Rice cultivars with xa-13 are resistant to prevalent races of bacteria] blight in the Indian subcontinent and should thus prove useful as donors for resistance to bacterial blight in rice breeding programs.     

Resistance to rice sheath blight (<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> Kühn) [(teleomorph: <Emphasis Type="Italic">Thanatephorus cucumeris</Emphasis> (A.B. Frank) Donk.] disease: current status and perspectives

Sheath blight (ShB) disease, caused by Rhizoctonia solani, is an economically important rice disease worldwide, especially in intensive production systems. Several studies have been conducted to identify sources for ShB resistance in different species of rice, including local accessions and landraces. To date, none of the genotypes screened are immune to ShB, although variation in levels of resistance have been reported. Several quantitative trait loci (QTL) for ShB resistance have been identified using mapping populations derived from indica or japonica rice. A total of 33 QTL associated with ShB resistance located on all 12 rice chromosomes have been reported, with ten of these co-localizing with QTL for morphological attributes, especially plant height, or for heading date. Sixteen QTL, from the same or differing genetic backgrounds, have been mapped at least twice. Of these, nine QTL were independent of morphological traits and heading date. We hypothesize that two main, distinct, mechanisms contribute to ShB resistance: physiological resistance and disease escape. Strategies to improve our understanding of the genetics of resistance to ShB are discussed.     

Late blight resistance in a diploid full-sib potato family

Late blight, caused by Phytophthora infestans (Mont.) de Bary, is the most destructive disease of potato worldwide. As this pathogen can rapidly overcome major race‐specific resistance genes, identifying the basis for enhanced quantitative resistance has become a crucial element for implementing advanced breeding strategies. A population of 230 full‐sib progeny derived from a cross between two diploid hybrid Solanum phureja × S. stenotomum clones was evaluated for foliage resistance against late blight in replicated trials at multiple locations in Pennsylvania between 1999 and 2002. In field experiments, plants were evaluated visually for per cent defoliation, and area under the disease progress curve (AUDPC) was determined. The two parents and three control cultivars (‘Atlantic’, ‘Kennebec’ and ‘Katahdin’) were included in all trials. In all three experiments, the presence of a significant number of clones exhibiting transgressive segregation were observed. There were significant differences among environments as well as among clones, and the clone × environment interaction was also significant. Stability analysis revealed that 37 clones made a significant contribution to the overall environment × clone interaction. Broad‐sense heritability for resistance, measured as AUDPC, was estimated as 0.67. The overall results indicate the presence in this potato family of a high level of field resistance against late blight. This segregating diploid family appears to be a good candidate for quantitative trait loci mapping to identify and characterize the genetic components of partial late blight resistance.     

Chromosomal location of Fusarium head blight resistance genes and analysis of the relationship between resistance to head blight and brown foot rot

In order to identify chromosomes involved in resistance to Fusarium head blight, a set of 21 substitution lines of Triticum macha (resistant) chromosomes into ‘Hobbit’'sib’(susceptible) were evaluated in trials over 2 years. For the first year's trial, all plants were inoculated on the same day with a conidial suspension of F. culmorum. For the second trial, individual plants were inoculated precisely at mid anthesis of each plant over a period of 2 weeks. The disease level was assessed by visual scoring, relative ear weight and F. culmorumn‐specfic quantitative polymerase chain reaction. The results showed that T. macha chromosomes 1B, 4A and 7A conferred good overall resistance, suggesting that they carry important genes for resistance. In two additional trials, T. macha and ‘Hobbit’'sib’ were evaluated for resistance to brown foot rot. The results showed that T. macha was more susceptible than ‘Hobbit’‘sib’, indicating that stem base disease response is not correlated with head blight resistance in these cultivars.     

水稻白叶枯病抗性研究进展

白叶枯病是世界水稻重要病害之一,且已成为研究植物和病原菌互作的模式,对该病的研究对其它病害有借鉴意义。目前已鉴定出29个抗白叶枯病基因,其中17个基因被定位到染色体上,4个基因已被克隆,在这些工作的基础上,已通过分子标记辅助选择和转基因方法育成了一些抗病新品系,展示了水稻抗白叶枯病分子育种的广阔前景。     

Breeding of transgenic rice restorer line for multiple resistance against bacterial blight, striped stem borer and herbicide

In this paper, we described the breeding of transgenic rice restorer line for multiple resistance against bacterial blight, striped stem borer (SSB) and herbicide by conventional crossing of two transgenic parental lines transformed independently with different genes. Two stable transgenic rice lines used as donor parents were developed, one was Zhongguo91 which contained cry1Ab gene (for insect resistance) and bar gene (for tolerance of herbicide), and the other was Yujing6 which contained Xa21 gene (resistance to bacterial blight). The elite restorer line Hui773 was used as recipient and crossed with the two stable transgenic rice lines. Then five successive backcrosses were made using Hui773 as recurrent parent. Two rice elite restorers, T773-1 expressing cry1Ab and bar genes and T773-2 expressing Xa21 gene, were obtained, which were confirmed by PCR analysis and testing selectable marker genes in the hybrid progenies. The cross was made between T773-1 and T773-2 to select stable restorer line carrying Xa21, cry1Ab and bar genes. Finally, we obtained transgenic restorer line T773 with good agronomic traits and obvious multiple resistance to Xanthomonas oryzae pv. oryzae, striped stem borer (Chilo suppressalis) and herbicide. The hybrid F₁ generation produced from the cross between transgenic restorer line T773 and a corresponding male sterile line Zaohua2A maintained obvious resistance to rice bacterial blight, rice leaffolder and striped stem borer, and showed significant heterosis. Our results indicate that it is feasible to develop transgenic hybrid rice cultivar through breeding transgenic restorer lines.     

Molecular characterization of <Emphasis Type="Italic">Fusarium</Emphasis> head blight resistance from wheat variety Wangshuibai

Fusarium head blight (FHB) is a destructive disease of wheat worldwide. FHB resistance genes from Sumai 3 and its derivatives such as Ning 7840 have been well characterized through molecular mapping. In this study, resistance genes in Wangshuibai, a Chinese landrace with high and stable FHB resistance, were analyzed through molecular mapping. A population of 104 F₂-derived F₇ recombinant inbred lines (RILs) was developed from the cross between resistant landrace Wangshuibai and susceptible variety Alondras. A total of 32 informative amplified fragment length polymorphism (AFLP) primer pairs (EcoRI/MseI) amplified 410 AFLP markers segregating among the RILs. Among them, 250 markers were mapped in 23 linkage groups covering a genetic distance of 2,430 cM. In addition, 90 simple sequence repeat (SSR) markers were integrated into the AFLP map. Fifteen markers associated with three quantitative trait loci (QTL) for FHB resistance (P < 0.01) were located on two chromosomes. One QTL was mapped on 1B and two others were mapped on 3B. One QTL on 3BS showed a major effect and explained up to 23.8% of the phenotypic variation for type II FHB resistance.     

Correlation between late blight resistance and foliage maturity type in potato

The genetics of race-non-specific foliage resistance against Phytophthora infestans, of foliage maturity type, and of their association in potato (Solanum tuberosum) were studied. Six progenies were derived from a half-diallel set of crosses between diploid potato clones that represented a broad pool within the genus Solanum and were free of any of the 11 known R genes for late blight resistance. The progenies were evaluated for resistance to late blight and for foliage maturity type, and five of them showed a significant correlation between the two traits. The correlation did not account for all variation that was present for both traits, as reflected in the analysis in which the relative AUDPC values were adjusted for foliage maturity type. The present study adds to previous results: resistance against P. infestans always coincides with late foliage maturity. However, the results also indicate that some selection for late blight resistance without affecting the foliage maturity type should be possible.     

Characterization and allelism of soybean mosaic virus strain SC3 resistance in ‘Qihuang-1’ derived soybean cultivars

Soybean mosaic virus is a severe constraint of soybean production in China. A total of country-wide 22 SMV strains (SC1-SC22) were identified. Of these, SC3 is a major strain widely distributed in Huanghuai and Yangtze River Valley region of China. Soybean cultivar ‘Qihuang-1’ contains R_SC3Q locus conditioning the resistance to SC3 and is an important parental line extensively used to breed the soybean cultivars in China. The objective of this study was to elucidate the genetic pattern of SC3 resistance genes in cultivars developed from ‘Qihuang-1’ or its derivative lines. Hence, we have evaluated the SC3 resistance in 91 cultivars developed from ‘Qihuang-1’ or its derivative lines. The results showed that a total of 43 cultivars exhibited resistance to the SC3 strain. Among them, 37 cultivars were derived from ‘Qihuang-1’. Then, we have detected the R_SC3Q loci in these cultivars using four SSR markers (Satt334, Sct_033, BARCSOYSSR_13_1114 and BARCSOYSSR_13_1136). It revealed that, among the 37 resistant cultivars derived from ‘Qihuang-1’, there are 20 cultivars containing R_SC3Q loci. Moreover, the allelic relationship of resistance genes was analysed using the crosses from resistance × resistance between ‘Qihuang-1’ and its resistant derived cultivars. The results showed that the resistance genes of ‘Qihuang-1’ and its 20 cultivars were allelic. But it is not allelic with those of the other 17 cultivars, different from ‘Qihuang-1’, and also, R_SC3Q does not condition the resistance. These results will be beneficial to exploring the transmission of resistance genes of ‘Qihuang-1’ and will be useful to the disease resistance breeding of soybean.     

Genetic mapping of bph20(t) and bph21(t) loci conferring brown planthopper resistance to Nilaparvata lugens St?l in rice (Oryza sativa L.)

Brown planthopper(BPH) is one of the most serious and destructive insect pests of rice in most rice growing regions of the world. In this study, two major resistance genes against BPH have been identified in an Oryza rufipogon (Griff.) introgression rice line, RBPH54. Inheritance of the BPH resistance in RBPH54 was studied by screening the resistance in parents, F1, F2 and BC1 generations against BPH biotype 2. A population of BC3F2 lines was developed and SSR markers were employed for the gene mapping, and new markers were designed for fine mapping of the resistance genes, while sequence information of BAC/PAC clones was used to construct physical maps of the genes. The results showed that the BPH resistance in RBPH54 was governed by recessive alleles at two loci, tentatively designated as bph20(t) and bph21(t). The locus bph20(t) was fine mapped to the short arm of chromosome 6 about 2.7 cM to the upper marker RM435 and 2.5 cM to lower marker RM540 and in a 2.5 cM region flanked by two new SSR markers BYL7 and BYL8 which were developed in the present study. The other BPH resistance locus bph21(t) was initially mapped to a region 7.9 cM to upper marker RM222 and 4.0 cM to lower marker RM244 on the short arm of chromosome 10. For physical mapping, the bph20(t)-linked markers were landed on BAC/PAC clones of the reference cv., Nipponbare, released by the International Rice Genome Sequencing Project. The bph20(t) locus was physically defined to an interval of about 75 kb with clone P0514G1. Identification and location of these two genes in the present study have diversified the BPH resistance gene pool, which give benefit to the development of resistant rice cultivars, and the linkage PCR-based SSR markers for the bph20(t) and bph21(t) genes would help realize the application of the genes in rice breeding through marker-assisted selection.     

Cloning, genetic and physical mapping of resistance gene analogs in barley (Hordeum vulgare L.)

The majority of verified plant disease resistance genes isolated to date belong to the NBS‐LRR class, encoding proteins with a predicted nucleotide binding site (NBS) and a leucine‐rich repeat (LRR) region. Using degenerate primers, designed from the conserved motifs of the NBS region in tobacco N and Arabidopsis RPS2 genes, we isolated 190 resistance gene analogs (RGA) clones from barley genomic DNA. A total of 13 single‐ and low‐copy RGAs were genetically mapped onto chromosomes 1H–7H (except 5H) using three barley double haploid (DH) mapping populations: Steptoe × Morex, Harrington × TR306 and LUGC × Bowman. Sequence analysis of the RGAs showed that they are members of a diverse group. As a result of BLAST searches, one RGA proved unique as it did not detect any significant hit. Another RGA is putatively functional, because it detected several barley expressed sequence tag (EST) matches. To physically map the RGAs, 13 sequences were used to screen a 6.3 × cv. ‘Morex’ bacterial artificial chromosome (BAC) library. After fingerprint analysis, eight contigs were constructed incorporating 62 BAC clones. These BAC contigs are of great value for positional cloning of disease resistance genes, because they span the regions where various barley R genes have been genetically mapped.     

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.     

QTL analysis of cooked rice grain elongation, volume expansion, and water absorption using a recombinant inbred population

The traits of elongation, volume expansion, and water absorption are very important in determining the quality of cooked rice grains. In this study, quantitative trait loci (QTL) analysis of these traits was performed using a recombinant inbred population derived from a cross between two indica cultivars, ‘Zhenshan 97’ and ‘Minghui 63 ,’ which are the parents of the most widely grown hybrid rice in China. Using a linkage map based on 221 molecular marker loci covering a total of 1796 cM, a total of 33 QTLs were identified for the nine traits tested. QTLs were detected on chromosomes 1– 3 , 5– 9 , and 11 , respectively. The QTLs identified included three for cooked rice grain length elongation (chromosomes 2 , 6 , and 11), six for width expansion (chromosomes 1‐ 3 , 6 , 9 , and 11) and two for water absorption (chromosomes 2 and 6). Interestingly, a single QTL located near the wx gene on chromosome 6 seemed to influence all the traits tested for the cooked rice quality.     

Substitution mapping of QTLs for blast resistance with SSSLs in rice (Oryza sativa L.)

Rice blast, caused by fungal pathogen Magnaporthe grisea Barr., is one of the most devastating rice diseases worldwide. It has greatly affected the rice production and quality. Development of resistant cultivars is the most effective and economical way for controlling this disease in rice. In this study, 114 of the single-segment substitution lines (SSSLs) in rice were inoculated at seedling stage by 16 rice blast isolates. The substituted segments in the 114 SSSLs distributed on 12 chromosomes with coverage of 57.32% of rice genome. Fifteen of the SSSLs were different in blast resistance from the HJX74 recipient. The SSSL W23-7-6-5-2-2 showed 100% of resistance frequency. A total of 11 QTLs for blast resistance were detected on chromosomes 1, 2, 3, 6, 10, 11 and 12 in rice. They were mapped at chromosomal intervals of 2.2?C46.2?cM, of which 6 QTLs were mapped at less than 10.0?cM. Six of the 11 QTLs were first reported in this paper.