小麦抗白粉病近等基因系遗传背景的分子标记检测

利用AFLP分子标记技术,对近年来培育的几套近等基因系的遗传背景进行了检测,发现所检测到的遗传多样性主要集中于供体亲本之间,而近等基因系内部已达到很高的遗传一致性;包含Pm4b的近等基因系、包含Pm2/Pm6的近等基因系与轮回亲本遗传上比较接近,其内部选系之间也达到很高的一致性,是理想的近等基因系;发现Pm21中簇毛     

STMS markers for grain protein content and their validation using near-isogenic lines in bread wheat

The present study, undertaken as a continuation of an earlier study on quantitative trait loci (QTL) analysis of grain protein content (GPC) in bread wheat (Prasad et al. 1999), includes the following: (1) identification of an additional molecular marker associated with GPC; (2) development of near‐isogenic lines (NILs) for high GPC; and (3) the use of three sets of NILs (a total of 10 NILs) to validate the two available markers linked with QTL for GPC. A total of 114 sequence‐tagged microsatellite site (STMS) primer pairs (that were not used in the previous study) were used for detection of polymorphism between the two parents (PH132, with high GPC; WL711, with low GPC) of a mapping population of 100 recombinant inbred lines (RILs). A total of 95 primer pairs gave amplification products, of which only 30 detected reproducible polymorphism between the parental genotypes. Bulked segregant analysis was conducted using these 30 primers on two bulks (each comprising eight RILs) representing the two extremes of the normal distribution. A solitary primer pair (WMC415) showed association with GPC, which was further confirmed through selective genotyping. Subsequently, 100 RILs were genotyped. A single‐marker linear regression analysis showed significant association between the marker WMC415 and GPC, thus identifying a quantitative trait locus (designated as QGpcccsu‐5A1), which explained 6.21% of the variation for GPC among the RILs. The above STMS marker, together with the STMS marker (WMC41) identified earlier, explains approximately 25% of the variation for GPC. In order to conduct validation of the above two available markers, 10 NILs were developed for high GPC using two genotypes (WL711 and HD2329) with low GPC as recipient parents and another two genotypes (PH132 and PH133) with high GPC as donor parents. NIL 2233 (with 11.7% GPC), derived from HD2329, when tried with WMC41 gave a characteristic amplification profile similar to that of its donor parent PH132, and NIL 2215 (with 11.9% GPC) derived from WL711, when tried with WMC415 gave an amplification profile that resembled its donor parent PH133. The remaining eight NILs with high GPC gave patterns similar to those of their corresponding recipient parents with both the markers, suggesting that either the QTL, other than those associated with the above markers, were actually transferred from the donor parents and contributed to high GPC in these NILs or that recombination had occurred between the markers identified and the corresponding QTL. Thus, the marker validation conducted using NILs, while demonstrating the utility of these two microsatellite markers for use in marker‐assisted selection in plant breeding, also suggested that many more QTL exist that would need to be identified using closely linked molecular markers.     

Effect of three anthocyaninless genes on germination in tomato (Lycopersicon esculentum Mill.) II. Seed germination under stress conditions

Seeds from four pairs of tomato isogenic/near-isogenic lines (IL/NIL) differing for anthocyaninless of Hoffmann (ah), three pairs of IL/NILs differing for anthocyaninwithout (aw) and six pairs of IL/NILs differing for baby lea syndrome (bls) were evaluated for their germination ability under stress conditions: low and high temperature, salt and osmotic stress. Mutant genes affecting anthocyanin biosynthesis enhanced tomato germination capacity under all above mentioned treatments, except for the aw gene under a PEG-6000 treatment. Testa removal accelerated germination under stress conditions, but differences in time to 50% germination between the wild type lines and their ah-, aw- and bls-IL/NILs although diminished, were not eliminated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of row type,flowering type and several other spike characters on resistance to Fusarium head blight in barley

Fusarium head blight (FHB) is a destructive disease of barley. The genetics and expression of resistance to FHB in barley is complex, and various spike characters are thought to possibly influence resistance. Tests using spray-inoculation of Fusarium graminearum at anthesis in greenhouse environments showed that two-rowed and cleistogamous varieties from Japan belong to the highest resistance group, while six-rowed and chasmogamous varieties are mostly susceptible. In order to evaluate the effect of such spike characters, including row type and flowering type, on FHB resistance, near-isogenic lines (NILs) differing in these characters were tested for their resistance. Two testing methods were used: the pot-plant and cut-spike methods, in which spikes at anthesis were spray-inoculated in greenhouse environments. The chasmogamous NILs and some six-rowed NILs were significantly more diseased than cleistogamous and two-rowed parent lines, respectively, and the difference in FHB severity was greater and more stable between cleistogamous/chasmogamous NIL pairs than between two-/six-rowed pairs. Slight or no differences were observed in glaucous/non-glaucous, normal/dense spike, normal/uzu type and normal/deficiens NIL pairs. The results indicate that the contribution of cleistogamy and/or the genetic background toward FHB resistance is more than that of row type and the other tested spike characters. Further, it should be possible to develop six-rowed varieties with FHB resistance nearly as good as that of the two-rowed varieties.     

Climacteric and non-climacteric behavior in melon fruit: 2. Linking climacteric pattern and main postharvest disorders and decay in a set of near-isogenic lines

A set of near-isogenic lines (NILs) of melon (Cucumis melo L.) was used to test the relationship between the climacteric pattern and postharvest disorders at harvest and after 30 days at 8 °C. The NILs contained different chromosome introgressions in the linkage group III from the non-climacteric exotic Korean accession PI 161375 transferred into the genetic background of the non-climacteric Spanish cultivar ‘Piel de Sapo’ (PS). A quantitative trait locus (QTL) in this linkage group induced climacteric behavior in eight NILs accompanied by a peak of ethylene production and fruit dehiscence to different degrees. The cultivar ‘Nicolás’ and one NIL showed a non-climacteric pattern of respiration rate and ethylene production. The climacteric NILs were used to test the relationship between this pattern and postharvest disorders. The reference climacteric lines ‘Fado’ and ‘Védrantais’ were more sensitive to CI and associated Cladosporium rot than the NILs or PS. In general, a more intense climacteric behavior was accompanied by fruit dehiscence, and higher total losses and greater skin scald after storage, than in PS. A higher incidence of chilling injury (CI) in the climacteric NILs was found compared with the non-climacteric ones, although with exceptions (one NIL for CI in the form of scald; the same NIL and one more for pitting). The climacteric onset and netting scald were not related, and CI in the form of skin spots was only found in climacteric NILs and was positively correlated with the maximum peak of ethylene production. Some climacteric NILs did not follow the rule of a higher susceptibility to other disorders and decay after storage compared with PS, such as for example in fruit over-ripening (detected externally or internally), Cladosporium rot at the peduncle and Alternaria rot. Mealiness was independent of climacteric behavior. Three climacteric NILs obtained better flavor scores after storage than PS, although the maximum peak of ethylene production was positively correlated with off-flavor. Genotypic correlation between disorder data and the physiological data of climacteric fruit revealed positive (flavor index) or negative postharvest consequences (skin injuries, rots or off-flavors). At least one QTL can be assigned to most of the quality traits analyzed.     

Confirmation of quantitative trait loci for resistance to multiple-HG types of soybean cyst nematode (<Emphasis Type="Italic">Heterodera glycines</Emphasis> Ichinohe)

Genetic analysis of resistance of plant introduction (PI) 438489B to soybean cyst nematode (SCN) have shown that this PI is highly resistant to many SCN HG types. However, validation of the previously detected quantitative trait loci (QTL) has not been done. In this study, 250 F_2:3 progeny of a Magellan (susceptible) × PI 438489B (resistant) cross were used for primary genetic mapping to detect putative QTL for resistance to five SCN HG types. QTL confirmation study was subsequently conducted using F_6:7 recombinant inbred lines (RILs) derived from the same cross. Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers were employed for molecular genotyping. Interval mapping (IM), permutation tests, cofactor selection, and composite interval mapping (CIM) were performed to identify and map QTL. Results showed that five QTL intervals were associated with resistance to either multiple- or single-HG types of SCN. Among these, two major QTL for resistance to multiple-SCN HG types were mapped to chromosomes (Chr.) 8 and 18, consistent with the known rhg1 and Rhg4 locations. The other QTL were mapped to Chr. 4. The results of our study confirmed earlier reported SCN resistance QTL in this PI. Moreover, SSR and SNP molecular markers tightly linked to these QTL can be useful for the near-isogenic lines (NILs) development aiming to fine-mapping of these QTL regions and map-based cloning of SCN resistance candidate genes.     

Fine mapping of a major quantitative trait locus, <Emphasis Type="Italic">qFLL6.2</Emphasis>, controlling flag leaf length and yield traits in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Fine mapping of a quantitative trait locus, qFLL6.2, controlling flag leaf length (FLL) and yield traits in rice was conducted using four sets of near isogenic lines (NILs) that were developed from a common residual heterozygote at F₇ generation of the indica rice cross Zhenshan 97/Milyang 46. Each of the NIL sets consisted of 40 lines that are S₁ progenies of ten maternal homozygotes, ten paternal homozygotes, and 20 heterozygotes differing in a portion of the 1.19-Mb interval RM3414–RM6917 on the short arm of rice chromosome 6. Analysis of phenotypic differences among the three genotypic groups in each NIL set delimited qFLL6.2 to a 62.1-kb region flanked by simple sequence repeat marker RM3414 and sequence-tagged site marker Si2944. This QTL explained 52.73% of the phenotypic variance, and the Zhenshan 97 allele increased FLL by 2.40 cm. Based on data collected from homozygous lines of three of the NIL sets, qFLL6.2 was shown to have major effects on all the three yield traits analyzed, including the number of spikelets per panicle, the number of filled grains per panicle, and grain weight per panicle. A comparison of the different groups revealed that the effect of qFLL6.2 was highly consistent across different genetic backgrounds and environments, providing a good candidate for map-based cloning and investigating the source–sink relationship in rice.     

Effect of the gene for double pod in chickpea on yield, yield components and stability of yield

Five pairs of near isogenic lines (NILs) for the gene for single vs. double pod were used to study the effect of this character on yield and its components (seeds per plant, yield per plant and seed weight). The NILs were sown in 2 years at five locations in ihe first year and four locations in the second. Combined analysis of variance was applied to t he data and no significant differences between isolines were found, but there were significant genotype– location interaction effects for yield and seeds per plant. For both characters the double–podded lines were more stable than the single–podded lines. Although there were significant differences between the five genotypes lor yield per planl and seed weight, the differentes between single– and double–podded lines were almost nil. The results indicate that the double–pod gene has a positive effect on the stability of yield and is not linked to any other gene responsible Tor seed size in chickpea     

玉米衔接式单片段导入系群体的构建和评价

以生产上广泛种植的玉米杂交种豫玉22的亲本自交系87-1和综3为受体亲本,以糯质、抗病性较好的玉米自交系衡白522为供体亲本,采用回交和自交的方法,结合SSR分子标记辅助选择,分别构建了以87-1和综3为背景的衔接式玉米单片段导入系群体,并对其遗传背景、导入片段大小、数目和覆盖率等进行了评价。结果表明,87-1背景的导入系群体导入了40个供体片段,片段长度介于0.03~342.8 cM,平均长度为91.1 cM,导入片段总长为3 643.9 cM,覆盖率为48.9%;综3背景的导入系群体导入了78个供体片段,片段长度介于0.03~343.4 cM,平均长度为75.5 cM,导入片段总长为5 895.2 cM,覆盖率为79.2%。     

Resistance to Fusarium crown rot in wheat and barley: a review

Fusarium crown rot (FCR) is becoming a major disease in many parts of the cereal‐growing regions worldwide. Significant QTL conferring FCR resistance have been reported on 13 of the 21 possible hexaploid wheat chromosomes in wheat and on three of the seven chromosomes in barley. Available results show that host resistance to FCR is not pathogen species‐specific, that resistance QTL have strong additive effect and that both plant height and growth rate affect FCR severity. Further, different loci seem to be responsible for resistances to FCR and Fusarium head blight although both diseases can be caused by the same Fusarium pathogens. Although marker‐assisted selection for FCR resistance has been initiated, the available markers are all derived from QTL mapping, which provides only limited resolution. Further work has to be conducted in developing diagnostic markers before significant progress can be made in deploying marker‐assisted selection as a routine tool to accelerate and improve FCR in breeding programmes.     

Yield and quality evaluations on a pair of processing tomato lines nearly isogenic for the Tm2a gene for resistance to the tobacco mosaic virus

A pair of processing tomato (Lycopersicon esculentum Mill.) lines, nearly isogenic for the Tm-2^a gene for resistance to tobacco mosaic virus, were grown in replicated trials under commercial production conditions in five locations worldwide. The lines were evaluated for 17 processing traits including fruit yield, size, soluble solids concentration, color, firmness, and viscosity. Eight of those traits differed significantly among the nearly-isogenic lines (NILs). Most notably, the NIL heterozygous for Tm-2^a yielded, on average, 16% more than the NIL homozygous for the susceptible allele and 33% more than the NIL homozygous for Tm-2^a. Viscosity was lower in the heterozygous NIL and the homozygous and heterozygous resistant NILs had softer fruit with larger stem scars compared to the homozygous susceptible NIL. These results indicate the presence of the Tm-2^a locus affects many traits of importance for processing tomatoes and may be used, in the heterozygous state, to significantly increase yield. Whether the observed effects are due to the Tm-2^a gene itself or genes associated via linkage drag could not be determined. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and validation of a major QTL for salt tolerance in soybean

To identify quantitative trait loci (QTLs) conditioning salt tolerance in soybean (Glycine max (L.) Merr.), two recombinant inbred line (RIL) populations derived from crosses of FT-Abyara × C01 and Jin dou No. 6 × 0197 were used in this study. The FT-Abyara × C01 population consisted of 96 F₇ RILs, and the Jin dou No. 6 × 0197 population included 81 F₆ RILs. The salt tolerant parents FT-Abyara and Jin dou No. 6 were originally from Brazil and China, respectively. The QTL analysis identified a major salt-tolerant QTL in molecular linkage group N, which accounted for 44.0 and 47.1% of the total variation for salt tolerance, in the two populations. In the FT-Abyara × C01 population, three RILs were found to be heterozygous around the detected QTL region. By selfing the three residual heterozygous lines, three sets of near isogenic lines (NILs) for salt tolerance were developed. An evaluation of salt tolerance of the NILs revealed that all the lines with FT-Abyara chromosome segment at the QTL region showed significantly higher salt tolerance than the lines without the FT-Abyara chromosome segment. Results of the NILs validated the salt tolerance QTL detected in the RIL populations.     

SCAR marker tagged to the alien leaf rust resistance gene Lr19 uniquely marking the Agropyron elongatum-derived gene Lr24 in wheat: a revision

A sequence characterized amplified region (SCAR) marker tagged to an Agropyron elongatum‐derived leaf rust resistance (Lr) gene Lr19 was validated on 18 known alien Lr gener in near‐isogenic lines (NILs) in the variety ‘Thatcher’, along with three wheat cultivers carrying Lr24 and two carrying Lr19. The marker was expressed only in the Lr24 lines confirming that the marker tagged the geneLr24. The monomorphic expression of the SCAR marker in 10NIL pairs for Lr19 and Lr24 revealed that each NIL pair possessed the same gene, Lr24. The donor parents used in the NIL pairs for Lr19 (‘Sunstar*6/C80‐1′) and Lr24 (‘TR380‐14*7/3Ag#14′) amplified the same fragment. Nonsegregation for leaf rust in the F₂ population of the cross between the above donor parents confirmed the presence of the same gene in the two parents. Apparently, a genuine parent stock of ‘Sunstar*6/C80‐1’ was not involved in the development of the NIL pairs for Lr19 due to an improper maintence bredding protocol either at source or destination which went undetected in the absence of signs of virulence for either gene in the region.     

Identification of resistance gene analogs as markers of disease resistance loci in oats, using near-isogenic lines

Genomic sequences with features of the major class of disease resistance genes and which bear nucleotide‐binding leucine‐rich repeat sequences (resistance gene analogs; RGA) were tested as potential markers of crown rust resistance loci in hexaploid oats. Two collections of paired near‐isogenic lines carrying resistance to different isolates of crown rust, Puccinia coronata were screened. Two out of the four RGAs assayed showed restriction fragment length polymorphism (RFLP) between one line of each collection and its recurrent parent. The paired lines X466 and D494 were polymorphic for RGA III2.2 and the pair of lines X470 and D504 were polymorphic for RGA III2.18. The III2.18 polymorphism was located in the hexaploid map Avena byzantina cv. ‘Kanota’ × A. sativa cv. ‘Ogle’ in linkage group KO17 in a region previously associated with crown rust resistance. In addition, 220 random primers were used for random amplified polymorphic DNA (RAPD) analysis to screen the two sets of NILs. Only one polymorphic band was obtained that differentiated the paired lines X470 and D504 from their parents. The RAPD band was used as a probe and the relevant RFLP that differentiated the NILs X470 and D504 was found at 1.7 cM from the III2.18 marker in KO17. RFLP analysis using probes previously mapped in KO17 confirmed differences for X470 and D504 in the region around the III2.18 marker. These results suggest that the resistance locus shared by this pair of NILs is probably linked to the markers revealed by RGA III2.18. The use of RGAs as RFLP probes in the screening of NILs with differences in crown rust resistance has proved to be more effective than RAPDs for finding polymorphic markers possibly linked to resistance loci.     

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。     

Effect of three anthocyaninless genes on germination in tomato (Lycopersicon esculentum Mill.) I. Seed germination under optimal conditions

The effect of three mutant genes affecting anthocyanin biosynthesis – anthocyaninless of Hoffmann (ah), anthocyaninwithout (aw) and baby lea syndrome (bls) – on tomato germination capacities at 24 ° C was investigated. The study was performed on nine anthocyanin containing lines – Apedice, Apeca, Ailsa Craig, Monfavet 167, Monfavet 168, Por, Piernita, VF 36 and VFNT cherry and 13 isogenic/near isogenic lines (IL/NIL) from them: 4 differing for gene ah, 3 differing for gene aw and 6 differing for gene bls). In the majority of the anthocyaninless IL/NILs germination began earlier than that in the wild type lines. Significant differences were observed in the time to 50% germination between the anthocyanin-containing and the anthocyaninless IL/NILs except for the most rapidly germinating line VFNT cherry. Seed weight, water uptake, effect of testa removal and presence or absence of maternal effects in F1 germination responses were investigated in order to elucidate the causes of the more rapid germination in the anthocyaninless IL/NILs. The increased amount of water uptake by the anthocyaninless lines, the shorter treatment time necessary for their testa removal, the important maternal effects in the genetic variability of the time to 50% germination and the differences in germination capacities between the wild type and the mutant IL/NILs after testa removal, indicated that the seed coat and the endosperm had to be affected by the effects of the three genes in a way that caused enhanced germination. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detecting major QTL associated with resistance to bacterial blight using a set of rice reciprocal introgression lines with high density SNP markers

Bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is a worldwide rice disease. QTL providing BB resistance were identified using a set of introgression lines (ILs) derived from a cross between Teqing and Lemont, and 3924 evenly distributed SNP markers developed from the two parents. After inoculating three Xoo pathotypes, CII, CIV and CV, seven major BB resistance‐associated QTL were detected. The alleles at all loci with improved BB resistance were from the Teqing background. Four QTL resistance to CII and CIV were identified in the reciprocal backgrounds across 2 years. Six QTL conferring resistance to CV identified in Teqing background were not detected in Lemont background, indicating that genetic background had a strong effect on the BB resistance‐associated QTL. Based on the interactions of 27 significant digenic QTL pairs among the seven main‐effect QTL, the QTL were divided into three strain‐specific groups. Genotype analyses of resistant ILs suggest that rice lines with a high level of resistance to BB can be achieved by pyramiding R gene(s) and QTL that interact with R gene(s).     

QTL mapping of a partial resistance to the corn delphacid‐transmitted viruses in Lepidopteran‐resistant maize line Mp705

A partial resistance to maize mosaic virus (MMV) and maize stripe virus (MStV) was mapped in a RILs population derived from a cross between lines MP705 (resistant) and B73 (susceptible). A genetic map constructed from 131 SSR markers spanned 1399 cM with an average distance of 9.6 cM. A total of 10 QTL were detected for resistance to MMV and MStV, using composite interval mapping. A major QTL explaining 34–41% of the phenotypic variance for early resistance to MMV was detected on chromosome 1. Another major QTL explaining up to 30% of the phenotypic variation for all traits of resistance to MStV was detected in the centromeric region of chromosome 3 (3.05 bin). After adding supplementary SSR markers, this region was found to correspond well to the one where a QTL of resistance to MStV already was located in a previous mapping study using an F₂ population derived from a cross between Rev81 and B73. These results suggested that these QTL of resistance to MStV detected on chromosome 3 could be allelic in maize genome.     

Development and Validation of SCAR Markers Co-Segregating with an Agropyron Elongatum Derived Leaf Rust Resistance Gene Lr24 in Wheat

Summary An Agropyron elongatum-derived leaf rust resistance gene Lr24 located on chromosome 3DL of wheat was tagged with six random amplified polymorphic DNA (RAPD) markers which co-segregated with the gene. The markers were identified in homozygous resistant F₂ plants taken from a population segregating for leaf rust resistance generated from a cross between two near-isogenic lines (NILs) differing only for Lr24. Phenotyping was done by inoculating the plants with pathotype 77-5 of Puccinia triticina. To enable gene-specific selection, three RAPD markers (S1302₆₀₉, S1326₆₁₅ and OPAB-1₃₈₈) were successfully converted to polymorphic sequence characterized amplified region (SCAR) markers, amplifying only the critical DNA fragments co-segregating with Lr24. The SCAR markers were validated for specificity to the gene Lr24 in wheat NILs possessing Lr24 in 10 additional genetic backgrounds including the Thatcher NIL, but not to 43 Thatcher NILs possessing designated leaf rust resistance genes other than Lr24. This indicated the potential usefulness of these SCAR markers in marker assisted selection (MAS) and for pyramiding leaf rust resistance genes in wheat.     

Quantitative trait loci associated with lodging tolerance in soybean cultivar ‘Toyoharuka’

Lodging tolerance (LT) is an important trait for high yield and combine-harvesting efficiency in soybean [Glycine max (L.) Merr.]. Many previous studies have investigated quantitative trait loci (QTLs) for lodging score (LS) in soybean. Most of the investigated QTLs were located in the proximal region of maturity or growth habit loci. The aim of this study was to identify genetic factors for LT not associated with maturity or growth habit. QTL analysis was performed using a recombinant inbred line (RIL) population derived from a cross between ‘Toyoharuka’ (TH), a lodging-tolerant cultivar, and ‘Toyomusume’ (TM). The genotypes of TH and TM were estimated as both e1e2E3E4 and dt1. The average LS over 4 years was used for QTL analysis, identifying a major and stable QTL, qLS19-1, on chromosome 19. The LS of the near-isogenic line (NIL) with the TH allele at Sat_099, the nearest marker to qLS19-1, was significantly lower than the NIL with the TM allele at that position. The TH allele at Sat_099 rarely had a negative influence on seed yield or other agronomic traits in both NILs and the TM-backcrossed lines. Our results suggest that marker-assisted selection for qLS19-1 is effective for improving LT in breeding programs.