Identification of QTL controlling adventitious root formation during flooding conditions in teosinte (Zea mays ssp. huehuetenangensis) seedlings

Adventitious root formation (ARF) at the soil surface is one of the most important adaptations to soil flooding or waterlogging. Quantitative trait loci (QTL) controlling ARF under flooding condition were identified in a 94 F₂ individual population by crossing maize (Zea mays L., B64) × teosinte (Z. mays ssp. huehuetenangensis). A base-map was constructed using 66 SSR and 42 AFLP markers, covering 1,378 cM throughout all ten maize chromosomes. The ARF capacity for seedlings was determined by evaluating the degree of root formation at the soil surface following flooding for 2 weeks. ARF showed continuous variation in the F₂ population. Interval mapping and composite interval mapping analyses revealed that the QTL for ARF was located on chromosome 8 (bin 8.05). Utilising a selective genotyping strategy with an additional 186 F₂ population derived from the same cross combination and 32 AFLP primer combinations, regions on chromosomes 4 (bin 4.07) and 8 (bin 8.03) were found to be associated with ARF. Z. mays ssp. huehuetenangensis contributed all of the QTL detected in this study. Results of the study suggest a potential for transferring waterlogging tolerance to maize from Z. mays ssp. huehuetenangensis.     

Molecular mapping of a new gene for resistance to frogeye leaf spot of soya bean in 'Peking'

Amplified fragment length polymorphism (AFLP) and microsatellite (simple sequence repeat, SSR) techniques were used to map the _RGSp_eking gene, which is resistant to most isolates of Cercospora sojina in the soya bean cultivar ‘Peking’. The mapping was conducted using a defined F₂ population derived from the cross of ‘Peking’(resistant) בLee’(susceptible). Of 64 EcoRI and MseI primer combinations, 30 produced polymorphisms between the two parents. The F₂ population, consisting of 116 individuals, was screened with the 30 AFLP primer pairs and three mapped SSR markers to detect markers possibly linked to Rcs_Peking. One AFLP marker amplified by primer pair E‐AAC/M‐CTA and one SSR marker Satt244 were identified to be linked to Res_Peking. The gene was located within a 2.1‐cM interval between markers AACCTA178 and Satt244, 1.1 cM from Satt244 and 1.0 cM from AACCTA178. Since the SSR markers Satt244 and Satt431 have been mapped to molecular linkage group (LG) J of soya bean, the Res_Peking resistance gene was putatively located on the LG J. This will provide soya bean breeders an opportunity to use these markers for marker‐assisted selection for frogeye leaf spot resistance in soya bean.     

Amplified fragment length polymorphism- and simple sequence repeat-based molecular tagging and mapping of greenbug resistance gene Gb3 in wheat

The greenbug, Schizaphis graminum (Rondani), is the most economically damaging aphid pest of wheat in the southern Great Plains of the USA. In this study, the single, dominant greenbug resistance gene, Gb3, was molecularly tagged and genetically mapped using amplified fragment length polymorphism (AFLP) and simple sequence repeat(SSR) markers. Three AFLP loci were associated with the Gb3 locus in linkage analysis with 75 F_2:3 families from the cross between two near‐isogenic lines (NILs) for Gb3,‘TXGBE273’ and ‘TXGBE281′. Two of these loci, XMgcc Pagg and Xmagg Patg cosegregate with Gb3 in the population analysed. Further analysis indicated that XMgcc Pagg and Xmagg Patg are specific for the Gb3 locus in diverse genetic backgrounds. Two SSR markers, Xgwm111 and Xgwm428 previously mapped in wheat chromosome 7D, were shown to be linked with Gb3, 22.5 cM and 33.1 cM from Gb3, respectively, in an F₂ population of ‘Largo’בTAM 107’, suggesting that Gb3 is located in the long arm of chromosome 7D. The two AFLP markers cosegregating with Gb3 are valuable tools in developing molecular markers for marker‐assisted selection of greenbug resistance in wheat breeding.     

Application of microsatellites from maize to teosinte and other relatives of maize

Teosinte comprises different Zea species (Zea mays, Zea diploperennis, Zea perennis, Zea luxurians) that can be crossed with cultivated maize (Z. mays ssp. mays). Nine microsatellites from maize were applied to different teosinte species in order to evaluate their usefulness in markerbased exploitation of these genetic resources. The same microsatellites were tested with rye, barley, and sorghum as potential molecular markers for these species. Almost all microsatellite × teosinte combinations yielded polymerase chain reaction (PCR) fragments in the range of cultivated maize. Using an F₂ population of a cross between maize inbred A188 and an individual of Zea mays ssp. mexicana, amplification products for maize and teosinte originated from the same genomic location for each of nine microsatellites investigated. PCR fragments of reduced intensity were generally obtained by applying maize microsatellites to rye, barley and sorghum. Polymorphisms among accessions within teosinte (sub)species occurred frequently. In contrast, no polymorphisms were obtained within rye, barley, and sorghum. Hence, application of maize microsatellites to teosinte for fingerprinting or marker-assisted introgression of genomic regions from teosinte into cultivated maize appears promising.     

玉米抗丝黑穗病QTL分析

以Mo17（抗）×黄早四（感）的F₂分离群体（191个单株）为作图群体,构建了含有84个SSR位点和48个AFLP位点的遗传连锁图谱,全长1 542.9 cM,平均图距11.7 cM。在吉林省公主岭和黑龙江省哈尔滨2个地点通过人工接种方法对184个相应的F₃家系（缺失7个）进行抗病鉴定。采用复合区间作图法对抗丝黑穗病数量性状位点（QTL）进行定位及遗传效应分析。在吉林公主岭地区检测到5个QTL,分别位于第1、2、3、8、9染色体上,解释的表型方差为10.0%~16.3%。在黑龙江哈尔滨地区也检测到5个QTL,分别位于第1、2、3、4、7染色体上,解释的表型方差为4.6%~13.4%。比较分析发现,两地一致在第2、3染色体上各检测到1个QTL,其中第2染色体上的表现为超显性效应,第3染色体上的表现为加性效应。研究结果为玉米抗丝黑穗病种质改良提供了重要信息。     

Construction of a high-density SSR marker-based linkage map of zoysiagrass (<Emphasis Type="Italic">Zoysia japonica</Emphasis> Steud.)

A consensus genetic linkage map with 447 SSR markers was constructed for zoysiagrass (Zoysia japonica Steud.), using 86 F₁ individuals from the cross ‘Muroran 2’ × ‘Tawarayama Kita 1’. The consensus map identified 22 linkage groups and had a total length of 2,009.9 cM, with an average map density of 4.8 cM. When compared with a previous AFLP-SSR linkage map, the SSR markers from each linkage group mapped to similar positions in both maps. Eight pairs of linkage groups from the AFLP-SSR map were joined into eight new groups in the current map. This zoysiagrass consensus map contained 35 SSR markers exhibiting high homology with rice genomic sequences from known chromosomal locations. This allowed synteny to be identified between Zoysiagrass linkage groups 2, 3, 9, 19 and rice chromosomes 3, 12, 2, 7 respectively. These results provide important comparative genomics information and the new map is now available for quantitative trait locus analysis, marker-assisted selection and breeding for important traits in zoysiagrass.     

QTL analysis of resistance to Fusarium head blight in wheat using a 'Frontana'-derived population

Fusarium head blight (FHB or head scab) has become a major limiting factor for sustainable wheat (Triticum aestivum L.) production around the world. For quantitative trait loci (QTL) analysis of resistance to FHB, F₃ plants and F_{3 : 5} lines, derived from a ‘Frontana’ (moderately resistant)/‘Seri82’ (susceptible) cross, were spray‐inoculated in 2001 and 2002, respectively. Artificial inoculations were carried out under field conditions. Of 273 SSR and AFLP markers, 250 could be mapped and they yielded 42 linkage groups, covering a genetic distance of 1931 cM. QTL analysis was based on the constructed linkage map and area under the disease progress curve (AUDPC). The analyses revealed three consistent QTLs associated with FHB resistance on chromosomes 1BL, 3AL and 7AS explaining 7.9%, 7.7% and 7.6% of the phenotypic variation, respectively, above 2 years. The results confirmed the previously described resistance QTL of ‘Frontana’ on chromosome 3AL. A combination of ‘Frontana’ resistance with ‘Sumai‐3’ resistance may lead to lines with augmented resistance expression.     

An Evaluation of Hordeum vulgare L. ×Psathyrostachys fragilis (Boiss.) Nevski Crosses for Doubled Haploid Barley Production

Investigations were carried out to assess the suitability of the intergeneric cross Hordeum vulgare×Psathyrostachys fragilis for haploid barley production. H. vulgare cvs. ‘Emir’ and ‘Vada’ were each pollinated with P. fragilis P.I. 343192 and plants regenerated from embryos cultured on a modified B5 medium. Seed sets on ‘Vada’ were significantly lower than on ‘Emir’, and all the planes from ‘Vada’×P. fragile remained hybrid. Several of these flowered but there was little pairing between the parental chromosomes. Most of the plants from ‘Emir’×P. fragilis died, as seedlings but 3 plants developed into haploid barley. Because of the practical limitations of pollen availability from P. fragilis and the inconsistencies in haploid plant formation, it is unlikely that the cross will prove as valuable as that between H. vulgare×H. bulbosum for a doubled haploid barley programme.     

QTL analysis of quality traits in the spring wheat cross RL4452 × 'AC Domain'

Wheat grain quality is a complex group of traits of tremendous importance to wheat producers, end‐users and breeders. Quantitative trait locus (QTL) analysis studied the genetics of milling, mixograph, farinograph, baking, starch and noodle colour traits in the spring wheat population RL4452/‘AC Domain’. Forty‐seven traits were measured on the population and 99 QTLs were detected over 18 chromosomes for 41 quality traits. Forty‐four of these QTLs mapped to three major QTL clusters on chromosomes 1B, 4D, and 7D. Fourteen QTLs mapped near Glu‐B1, 20 QTLs mapped near a major plant height QTL on chromosome 4D, and 10 QTLs mapped near a major time to maturity QTL on chromosome 7D. Large QTLs were detected for grain and flour protein content, farinograph absorption, mixograph parameters, and dietary fibre on chromosome 2BS. QTLs for yellow alkaline noodle colour parameter L* mapped to chromosomes 5B and 5D, while the largest QTL for the b* parameter mapped to 7AL.     

Development of molecular markers for crown rot resistance in wheat: mapping of QTLs for seedling resistance in a '2-49' × 'Janz' population

Crown rot, caused by Fusarium pseudograminearum, is an important disease of wheat in Australia and elsewhere. In order to identify molecular markers associated with partial seedling resistance to this disease, bulked segregant analysis and quantitative trait loci (QTL) mapping approaches were undertaken using a population of 145 doubled haploid lines constructed from ‘2‐49’ (partially resistant) × ‘Janz’ (susceptible) parents. Phenotypic data indicated that the trait is quantitatively inherited. The largest QTLs were located on chromosomes 1D and 1A, and explained 21% and 9% of the phenotypic variance, respectively. Using the best markers associated with five QTLs identified by composite interval mapping, the combined effect of the QTLs explained 40.6% of the phenotypic variance. All resistance alleles were inherited from ‘2‐49’ with the exception of a QTL on 2B, which was inherited from ‘Janz’. A minor QTL on 4B was loosely linked (19.8 cM) to the Rht1 locus in repulsion. None of the QTLs identified in this study were located in the same region as resistance QTLs identified in other populations segregating for Fusarium head blight, caused by Fusarium graminearum.     

Molecular mapping of genes involved in root hair formation in barley

In the presented study, the existing AFLP and SSR maps of barley were used to find chromosomal position of four genes controlling different stages of root hair development. Four barley mutants were used in the analysis: the root hairless mutant rhl1.b, mutant rhp1.b with root hair development blocked at the initial bulge formation, mutant rhi1.a with irregular pattern of sparsely located root hairs and mutant rhs1.a with very short root hairs. Each mutant was crossed with parents of ‘Steptoe’/‘Morex’ mapping population and F₂ progenies of crosses: mutant × ‘Steptoe’ and mutant × ‘Morex’ were analyzed for segregation of root hair phenotype and polymorphic AFLP and SSR markers. It was possible to map all the analyzed genes on barley chromosomes: rhl1 gene on the short arm of chromosome 7H, rhp1 gene on chromosome 1H, rhs1 locus in the pericentromeric region of chromosome 5H and rhi1 gene on the long arm of chromosome 6H. Subsequently, the Bulk Segregant Analysis and AFLP technique were used for saturation of the identified regions with new markers. The joint maps were constructed using as common points the SSR markers located in the target regions. Linkage maps of the regions around the four genes involved in the root hair formation in barley were composed of 8–11 markers and spanned over 16.1–49.0 cM. The distances between localized genes and the closest markers ranged from 1.0 to 3.8 cM. The identified chromosomal locations of genes can be used for their fine mapping and future map-based cloning.     

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.     

Genetics and identification of molecular markers linked to resistance to loose smut (<Emphasis Type="Italic">Ustilago tritici</Emphasis>) race T33 in durum wheat

This study was conducted to determine the genetic control of resistance to loose smut caused by Ustilago tritici race T33 in two durum recombinant inbred line populations (DT662 × D93213 and Sceptre × P9162-BJ08*B) and to identify molecular markers linked to the resistance. Resistance in both populations was controlled by single genes. Two SSR markers were linked with loose smut resistance in the Sceptre × P9162-BJ08*B population. In DT662 × D93213, two AFLP, two wheat SSRs and one SCAR markers were linked to resistance. The SCAR marker, 3.2 cM distal to the smut resistance locus (Utd1) on chromosome 5BS, accounted for up to 64% of the variability in disease reaction; the other markers were proximal to Utd1 at genetic distances ranging from 5.9 to 35.9 cM. SSR markers Xgwm234 and Xgwm443 segregated in both crosses suggesting a common resistance gene. The SCAR and SSR markers can be used effectively for marker assisted selection to incorporate loose smut resistance into durum cultivars.     

Relationship of intra- and interpopulation tropical maize single cross hybrid performance and genetic distances computed from AFLP and SSR markers

Two sets of tropical maize inbred lines, one derived from the BR-105 population and another derived from the BR-106 population, were assayed for Amplified Fragment Length Polymorphism's (AFLP) and for Simple Sequence Repeat (SSR), in order to investigate genetic distances among lines and their relationship to heterotic group assignment and single cross yield performance. Genetic distances were on average greater for interpopulation than intrapopulation crosses for both AFLP and SSR. Cluster analysis was in agreement with the original assignment for heterotic groups. Inbred line 16, derived from BR-106, was assigned to the BR-105 set, in agreement with single cross yield performance from intra- and interpopulation crosses. However, the same pattern was not observed for SSR where another two lines from BR-106 were also assigned to the BR-105 set. Correlation coefficients of genetic distances (GD) with F₁ grain yield and heterosis were high for BR-106 ×BR-106 crosses (0.91^** and 0.82^** for AFLP and SSR, respectively), moderate for BR-105 × BR-105 crosses (0.52^* for AFLP and SSR) and low for BR-105 × BR-106 crosses (0.29 and 0.16 for AFLP and SSR, respectively). The lower correlation at interpopulation level was due to the smaller range of GD caused probably by a previous selection for combining ability. General results showed that the AFLP molecular marker is efficient in assigning maize lines to heterotic groups and that AFLP-based GD is suitable for predicting the maize single cross performance for intrapopulation crosses of broad-based populations. The efficiency of SSR in assigning lines to heterotic groups and for predicting single cross performance was smaller than AFLP. This revised version was published online in July 2006 with corrections to the Cover Date.     

The first genetic linkage map of crape myrtle (Lagerstroemia) based on amplification fragment length polymorphisms and simple sequence repeats markers

Lagerstroemia (crape myrtle) are famous ornamental plants with large pyramidal racemes, long flower duration and diverse colours. Genetic maps provide an important genomic resource of basic and applied significance. A genetic linkage map was developed by genotyping 192 F₁ progeny from a cross between L. caudata (female) and L. indica (‘Xiang Xue Yun’) (male) with a combination of amplification fragment length polymorphisms (AFLP) and simple sequence repeats (SSR) markers in a double pseudo‐testcross mapping strategy. A total of 330 polymorphic loci consisting of 284 AFLPs and 46 SSRs showing Mendelian segregation were generated from 383 AFLP primer combinations and 150 SSR primers. The data were analysed using JoinMap 4.0 (evaluation version) to construct the linkage map. The map consisted of 20 linkage groups of 173 loci (160 AFLPs and 13 SSRs) covering 1162.1 cM with a mean distance of 10.69 cM between adjacent markers. The 20 linkage groups contained 2–49 loci and ranged in length from 7.38 to 163.57 cM. This map will serve as a framework for mapping QTLs and provide reference information for future molecular breeding work.     

Chromosomal assignment of AFLP markers in upland cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.)

In this research, we used two sets of cotton aneuploid (G. hirsutum × G. tomentosum and G. hirsutum × G. barbadense) plants to locate AFLP markers to chromosomes using deletion analysis method. Thirty-eight primer combinations were used to generate 608 polymorphic AFLP markers. A total of 98 AFLP markers were assigned to 22 different cotton chromosomes or chromosome arms. Of those assigned markers, 63.3% were assigned to A genome and 36.7% were assigned to D genome. A low rate (14.3%) of common markers were found between those assigned AFLP markers with the AFLP markers from an intraspecific cross population developed previous in our lab. Based on the 16 common markers, we were able to associate the 13 linkage groups previously identified in our lab to eight chromosomes. Further research will be carried out by using SSR markers with known location to associate unassigned linkage groups to chromosomes.     

Construction of an integrated genetic map based on maternal and paternal lineages of tea (Camellia sinensis)

Genetic study on important traits of tea is difficult because of its self-incompatibility in nature. Moreover, development of a new variety usually needs more than 20 years, since it takes many years from seedling to matured plants for trait investigation. Genetic map is an essential tool for genetic study and breeding. In this study, we have developed an integrated genetic map of tea (Camellia sinensis) using a segregating F1 population derived from a cross between two commercial cultivars (‘TTES 19’ and ‘TTES 8’). A total of 574 polymorphic markers (including SSR, CAPS, STS, AFLP, ISSR and RAPD), 69 markers with highly significant levels of segregation distortion (P < 0.001) (12.0 %) were excluded from further analyses. Of the 505 mapped markers, there were 265 paternal markers (52.5 %), 163 maternal markers (32.3 %), 65 doubly heterozygous dominant markers (12.9 %), and 12 co-dominant markers (2.4 %). The co-dominant markers and doubly heterozygous dominant markers were used as bridge loci for the integration of the paternal and maternal maps. The integrated map comprised 367 linked markers, including 36 SSR, 3 CAPS, 1 STS, 250 AFLP, 13 ISSR and 64 RAPD that were assigned to 18 linkage groups. The linkage groups represented a total map length of 4482.9 cM with a map density of 12.2 cM. This genetic map has the highest genetic coverage so far, which could be applied to comparative mapping, QTL mapping and marker assisted selection in the future.     

Molecular markers linked to <Emphasis Type="Italic">Bn</Emphasis>;<Emphasis Type="Italic">rf</Emphasis>: a recessive epistatic inhibitor gene of recessive genic male sterility in <Emphasis Type="Italic">Brassica napus </Emphasis>L.

7–7365AB is a recessive genic male sterile (RGMS) two-type line, which can be applied in a three-line system with the interim-maintainer, 7–7365C. Fertility of this system is controlled by two duplicate dominant epistatic genes (Bn;Ms3 and Bn;Ms4) and one recessive epistatic inhibitor gene (Bn;rf). Therefore an individual with the genotype of Bn;ms3ms3ms4ms4Rf_ exhibits male sterility, whereas, plant with Bn;ms3ms3ms4ms4rfrf shows fertility because homozygosity at the Bn;rf locus (Bn;rfrf) can inhibit the expression of two recessive male sterile genes in homozygous Bn;ms3ms3ms4ms4 plant. A cross of 7–7365A (Bn;ms3ms3ms4ms4RfRf) and 7–7365C (Bn;ms3ms3ms4ms4rfrf) can generate a complete male sterile population served as a mother line with restorer in alternative strips for the multiplication of hybrid seeds. In the present study, molecular mapping of the Bn;Rf gene was performed in a BC₁ population from the cross between 7–7365A and 7–7365C. Bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) technique was used to identify molecular markers linked to the gene of interest. From a survey of 768 primer combinations, seven AFLP markers were identified. The closest marker, XM5, was co-segregated with the Bn;Rf locus and successfully converted into a sequence characterized amplified region (SCAR) marker, designated as XSC5. Two flanking markers, XM3 and XM2, were 0.6 cM and 2.6 cM away from the target gene, respectively. XM1 was subsequently mapped on linkage group N7 using a doubled-haploid (DH) mapping population derived from the cross Tapidor × Ningyou7, available at IMSORB, UK. To further confirm the location of the Bn;Rf gene, additional simple sequence repeat (SSR) markers in linkage group N7 from the reference maps were screened in the BC₁ population. Two SSR markers, CB10594 and BRMS018, showed polymorphisms in our mapping population. The molecular markers found in the present study will facilitate the selection of interim-maintainer.     

玉米抗纹枯病QTL定位

以玉米自交系R15（抗）×掖478（感）的229个F₂单株为作图群体,构建了包含146个SSR标记位点的遗传连锁图谱,全长1 666 cM,平均图距11.4 cM。通过麦粒嵌入法对F2:4群体进行人工接种纹枯病菌,并以相对病斑高为病级划分标准鉴定了玉米纹枯病的抗性。用复合区间作图法分析抗病QTL及遗传效应,共检测到9个抗性QTL,分布于第1、2、3、4、5、6和10条染色体上,单个QTL可解释表型方差的3.72%~7.19%,其中有2个QTL位于染色体6.01抗病基因簇附近。     

Genomic in situ hybridization analysis of Brassica napus × Orychophragmus violaceus hybrids and production of B. napus aneuploids

To further utilize the valuable germplasm Orychophragmus violaceus for Brassica genetics and breeding, a B. napus × O. violaceus cross was repeated with embryo rescue. All F₁ plants except one B. napus haploid were mixoploids (2n = 17–39 in ovaries) with 2n = 31, 37, 38 and 39 as the maximal chromosome numbers in individuals, but the higher numbers mostly appeared in pollen mother cells (PMCs) with a preponderance of 2n = 30, 37 and 38. Only one chromosome and one chromosome segment of O. violaceus were detected at a low frequency in some ovary cells and PMCs with 2n = 37, 38 and 39 as determined by genomic in situ hybridization analysis. The fatty acid profiles of seeds from the majority of the F₁ and F₂ plants were similar to those of female B. napus cv. ‘Oro’, but some were obviously different in the percentages of oleic, linoleic and erucic acids, and some F₂ plants (2n = 38) with good seed set had high percentages of oleic (>70.0%) or linoleic (to 38.3%) acids and low erucic acid (<1%). Subsequently, many kinds of B. napus aneuploids (2n = 28, 30, 34, 36, 37, 39 and 42), without O. violaceus chromosomes, were derived from F₂ progeny and microspores of partial F₁ plants. Finally, the cytological mechanisms behind the variations in chromosome numbers were discussed together with the implications of these aneuploids for Brassica genome research and of the plants with altered fatty acid profiles for improving the oil quality of B. napus.