Molecular markers assist in the development of diverse inbred backcross lines in European Long cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.)

The popular fresh-market European Long cucumber (Cucumis sativus L.) is grown commercially worldwide under controlled, greenhouse environments. However, it has a narrow genetic base, where private and public improvement programs can trace their origins to comparatively few accessions. Therefore, a project was designed to identify diverse genotypes for use in the formation and analysis of inbred backcross (BC₂S₃) lines (IBL) to broaden the genetic base of this market class. Initially, 42 cucumber accessions were evaluated with a previously defined standard marker array to identify parents for use in backcrossing. The IBL were developed by crossing the elite commercial line NZ1 (Nunhems Vegetable Seeds, Haelen, The Netherlands) and PI 432858 (China), and then backcrossing the most genetically diverse BC₁ and BC₂ progeny to the elite parent as defined by marker analyses (19 polymorphic, mapped SSR, and SCAR marker loci), followed by three generations of single seed descent resulting in 116 IBL (BC₂S₃). The IBL were evaluated under greenhouse conditions for days to anthesis, sex expression, lateral branch number, yield, and exterior fruit quality in Madison, Wisconsin, USA (soil media), and in Haelen and Bergschenhoek, The Netherlands (soilless, hydroponic media). The IBL were genotyped using an expanded marker array (37 polymorphic SSR, SCAR, SNP, EST, BAC end, and gene-associated loci), and genetic relationships were examined by multivariate analyses using phenotypic and genotypic data. The 116 developed IBL possessed considerable morphological and genotypic diversity, where genetic distance (GD) among lines ranged between 0.00 and 0.77. These IBL possessed many commercially acceptable attributes, and, thus, genetic diversity in this market type could be substantially increased by the use of these genetically broad-based IBL during plant improvement.     

芥菜型油菜多室基因Bjmc2的精细定位

芥菜型多室油菜的产量比普通两室油菜更高,定位乃至克隆多室基因可为油菜遗传改良及解释多室角果形成机制创造条件。本研究通过验证JD11-2家系衍生群体仅在BjMc2位点上存在差异,可用于BjMc2的定位。采用AFLP结合BSA法分析BC₅和BC₆群体,筛选到1个与BjMc2连锁的AFLP标记并转化为SCAR标记SC1。基于该AFLP标记序列信息,利用白菜同源序列设计SSR引物和SCAR引物,获得11对SSR标记和1对SCAR标记。通过在芥菜型油菜BAC文库中的挑选,获得2个覆盖目标区域的单克隆,由此开发1个SSR标记。将获得的SCAR和SSR标记扫描BC₇群体,构建了两室性状基因BjMc2的遗传连锁图,两侧最近标记ZX17和BACsr96与目标基因之间的遗传距离分别为0.048 cM和0.340 cM,并定位到白菜A7 scaffold000019的946~1014 kb之间,约68 kb物理距离。     

Prospects for hybrid breeding in winter triticale: II. Relationship between parental genetic distance and specific combining ability

Significant relative midparent heterosis (MPH%) for grain yield in triticale (×Triticosecale Wittm.) has generated interest in the development of hybrid cultivars. The objectives of this study were to (i) examine the association between parental genetic distance (GD) and specific combining ability (SCA), (ii) investigate the existence of genetically distant heterotic groups in elite germplasm, and (iii) draw conclusions for future hybrid breeding in winter triticale. Genetic distance between 61 lines was estimated, based on 93 polymorphic simple sequence repeat (SSR) marker loci and 10 AFLP (amplified fragment length polymorphism) primer‐enzyme combinations (PEC). Agronomic data of 206 F₁ crosses and their 61 parental lines grown in six German environments were published recently in a companion study. Correlations were made between SCA for grain yield, number of spikes/m², 1000‐kernel weight and number of kernels per spike with GD estimates of the 56 female and five male parents (testers). Principal co‐ordinate analyses (PCoA) based on SSR data revealed no distinct subgroups in the germplasm. Correlations between GD and SCA were low for all traits (|r| ≤ 0.31), which hampers the prediction of SCA from molecular data. A multi‐stage procedure is recommended for future hybrid breeding in triticale by applying a pragmatic approach for the grouping of germplasm following the early history of hybrid breeding of maize.     

Comparative analysis of cultivated melon groups (Cucumis melo L.) using random amplified polymorphic DNA and simple sequence repeat markers

Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to characterize genetic relationships among 46 accessions in two C. melo L. subsp. melo (Cantalupensis, Inodorus) and subsp.agrestis (Conomon, and Flexuosus) groups. Genetic distance (GD) estimates were made among and between accessions in four melon market classes [Galia, Ogen, Charentais, and Shipper (European and U.S. types)] of Cantalupensis, one market class of Inodorus (Cassaba and Honey Dew), one accession of Conomon, and one accession of Flexuosus by employing three GD estimators; simple matching coefficient, Jaccard's coefficient, and Nei's distance-D. Differences detected among 135 RAPD bands and 54 SSR bands (products of 17 SSR primers) were used to calculate GD. Band polymorphisms observed with 21 RAPD primers and 7 SSR primers were important (p =0.01) in the detection of genetic differences. Estimators of GD were highly correlated (p 0.0001; r_s = 0.64 to0.99) when comparisons were made between estimation methods within a particular marker system. Lower correlations (r_s = 0.17 to 0.40) were detected (P > 0.001) between marker systems using any one estimator. The GD of the Conomon and Flexuosus accessions was significantly different (p> 0.001)from the mean GD of all the market classes examined. The mean GD (Jaccard's coefficient) among accessions of Ogen, Galia, Cassaba, Charentais, European shipper, and U.S. shipper groups was 0.11 ± 0.04, 0.33± 0.09, 0.21 ± 0.04, 0.26 ± 0.10, 0.17± 0.05 and 0.22 ± 0.08, respectively. Market classes were distinct (p > 0.001), such that GDs between Galia and other accessions were the largest(mean GD 0.34 to 0.35), and GDs between Ogen and other accessions were the smallest (mean GD 0.29 to 0.30). Contrasts between the U.S. shipper cultivar Top Mark and accessions within any market class was relatively large (mean GD = 0.42 ± 0.06). Empirical estimations of variances associated with each marker type in the accessions examined indicated that, per band, lower coefficients of variation can be attained in the estimation of GD when using RAPDs compared to SSRs. Nevertheless, the genetic relationships identified using these markers were generally similar. The disparity between the analyses of the two markers made may be related to the amount of genome coverage which is characteristic of a particular marker system and/or its efficiency in sampling variation in a population. Results of RAPD marker analysis suggest that 80 marker bands were adequate for assessing the genetic variation present in the accessions examined. This revised version was published online in July 2006 with corrections to the Cover Date.     

Use of molecular markers aids in the development of diverse inbred backcross lines in Beit Alpha cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.)

Beit Alpha cucumber (Cucumis sativus L.) is a Mediterranean fresh-market type with a relatively narrow genetic base. To broaden its base for plant improvement, 42 diverse accessions were compared employing a previously defined standard marker array to choose wide-based parental lines for use in backcross introgression. Inbred backcross lines (IBL) were developed by crossing Beit Alpha line ‘04HD5’ (De Ruiter Seeds, The Netherlands; recurrent parent) and PI 285606 (Poland; donor parent), and then selecting the most genetically diverse BC₁ and BC₂ progeny based on molecular marker profiles, followed by three generations of single-seed descent to produce 117 IBL. Molecular genotyping of IBL was then performed, and IBL were evaluated for days to anthesis, sex expression, pistillate flowers per node, lateral branch number, fruits per plant, fruit length, and fruit weight in the US, The Netherlands, Israel, and Turkey. Multivariate analyses and genetic distance comparisons indicate that IBL possessed considerable inter-line morphological and genotypic diversity. These diverse IBL will be useful in genetic studies and to evaluate Beit Alpha cross-progeny derived from IBL × elite germplasm created to broadened genetic base of this market type.     

Development of high yielding IR64 × <Emphasis Type="Italic">Oryza rufipogon</Emphasis> (Griff.) introgression lines and identification of introgressed alien chromosome segments using SSR markers

Modern rice varieties that ushered in the green revolution brought about dramatic increase in rice production worldwide but at the cost of genetic diversity at the farmers’ fields. The wild species germplasm can be used for broadening the genetic base and improving productivity. Mining of alleles at productivity QTL from related wild species under simultaneous backcrossing and evaluation, accompanied by molecular marker analysis has emerged as an effective plant breeding strategy for utilization of wild species germplasm. In the present study, a limited backcross strategy was used to introgress QTL associated with yield and yield components from Oryza rufipogon (acc. IRGC 105491) to cultivated rice, O. sativa cv IR64. A set of 12 BC₂F₆ progenies, selected from among more than 100 BC₂F₅ progenies were evaluated for yield and yield components. For plant height, days to 50% flowering and tillers/plant, the introgression lines did not show any significant change compared to the recurrent parent IR64. For yield, 9 of the 12 introgression lines showed significantly higher yield (19–38%) than the recurrent parent IR64. Four of these lines originating from a common lineage showed higher yield due to increase in grain weight and another three also from a common lineage showed yield increase due to increase in grain number per panicle. For analyzing the introgression at molecular level all the 12 lines were analyzed for 259 polymorphic SSR markers. Of the total 259 SSR markers analyzed, only 18 (7.0%) showed introgression from O. rufipogon for chromosomes 1, 2, 3, 5, 6 and 11. Graphical genotypes have been prepared for each line and association between the introgression regions and the traits that increased yield is reported. Based on marker trait association it appears that some of the QTL are stable across the environments and genetic backgrounds and can be exploited universally.     

Relationship between molecular marker heterozygosity and hybrid performance in intra- and interspecific hybrids of cotton

The present study was conducted to investigate the relationship between parental molecular marker diversity and hybrid performance in both intra‐ and interspecific hybrids of cotton to evaluate the feasibility of predicting hybrid performance using molecular markers. Three cytoplasmic male sterile (CMS) lines were crossed with 10 restorer lines to produce 22 F₁ hybrids during 2003. Of 22 F₁s, 14 hybrids were intraspecific (Gossypium hirsutum × G. hirsutum) and eight interspecific (G. hirsutum × G. barbadense). These 22 F₁ hybrids and their parents were evaluated for yield and fibre quality traits at Zhejiang University, Hangzhou, China during 2004 and 2005. Genetic distances (GD) among the parents were calculated from 56 random‐amplified polymorphic DNAs (RAPD) and 66 simple sequence repeat (SSR) marker data, and their correlation with hybrid performance and heterosis were analysed. The parents could be discriminated into G. hirsutum and G. barbadense clusters by cluster analysis based on both RAPD and SSR markers data. The correlation (r = 0.503, P ≤ 0.05) was calculated between GD_rapd (GD based on RAPD markers) and GD_ssr (GD based on SSR markers). Correlation of GD with hybrid performance and heterosis differed considerably between intra‐ and interspecific hybrids. The correlation between GD and hybrid performance was non‐significant for most of traits within the hybrids of G. hirsutum species. However, it was significantly and positively correlated for fibre length, fibre strength and elongation in interspecific hybrids. The relationship between GD and heterosis was observed to be positively significant for boll weight within hybrids of G. hirsutum with significant and negative correlations for fibre length and elongation. In conclusion, the power of predicting hybrid performance using molecular markers in cotton is low. But, the relationship between SSR marker heterozygosity and hybrid performance can be used to predict fibre length during interspecific hybrid cotton breeding.     

Identification of two SCAR markers co-segregated with the dominant Ms and recessive ms alleles in onion (Allium cepa L.)

Cytoplasmic genetic male-sterility is used to produce hybrid onion (Allium cepa L.) seeds worldwide. In this paper, we present the results of research aimed toward identifying PCR-based markers linked to the Ms locus through amplified fragment length polymorphism (AFLP). After screening 512 AFLP primer combinations, only one AFLP fragment was identified as being flanking linked to the dominant Ms allele. Subsequently, the AFLP marker was converted into a sequence-characterized amplified region (SCAR) marker, designated as DNF-566, co-segregated with the dominant Ms allele in first backcross (BC₁) segregated populations. Furthermore, we designed another molecular marker (RNS-357) co-segregated with the ms allele to identify different genotypes (i.e., MsMs, Msms, or msms). Both markers could be used for evaluating onion lines with different genetic backgrounds (including male-sterile lines, maintainer lines, male-fertile lines, and commercial based F₁ hybrid cultivars). The results of this study indicate that maintainer plants could be directly selected by using these 2 SCAR markers in the onion breeding process, and this may contribute significantly toward breeding onion F₁ hybrid cultivars.     

Persistent C genome chromosome regions identified by SSR analysis in backcross progenies between Brassica juncea and B. napus

Given that feral transgenic canola (Brassica napus) from spilled seeds has been found outside of farmer’s fields and that B. juncea is distributed worldwide, it is possible that introgression to B. juncea from B. napus has occurred. To investigate such introgression, we characterized the persistence of B. napus C genome chromosome (C-chromosome) regions in backcross progenies by B. napus C-chromosome specific simple sequence repeat (SSR) markers. We produced backcross progenies from B. juncea and F₁ hybrid of B. juncea × B. napus to evaluate persistence of C-chromosome region, and screened 83 markers from a set of reported C-chromosome specific SSR markers. Eighty-five percent of the SSR markers were deleted in the BC₁ obtained from B. juncea × F₁ hybrid, and this BC₁ exhibited a plant type like that of B. juncea. Most markers were deleted in BC₂ and BC₃ plants, with only two markers persisting in the BC₃. These results indicate a small possibility of persistence of C-chromosome regions in our backcross progenies. Knowledge about the persistence of B. napus C-chromosome regions in backcross progenies may contribute to shed light on gene introgression.     

Construction and characterization of 3-S Lines, an alternative population for mapping studies in rice (Oryza sativa L.)

Most traits of agronomic importance in rice are quantitative in nature and are controlled by polygenes, called quantitative trait loci (QTL). Understanding the nature and effect of QTLs are important for rice breeding to achieve higher yield and stability. Single segment substitution lines (SSSLs or 3-S Lines) were developed through simple sequence repeats (SSR) marker-facilitated backcrossing methods for Hua-Jing-Xian 74 (HJX74) with the donor segment from six elite germplasm and was characterized. Complete genome survey was carried out with 258 polymorphic SSR markers. Polymorphism of the donors with the recurrent parent varied between 32.98 and 60.73% with an average of 47.81%. Japonica donors were more polymorphic than indica donors. Number of substitution segments per plant decreased with the advancement of backcross generations. In BC₂F_1, BC₃F₁, BC₃F₂ and BC₃F₃ the average number of substitution segment per plant were 12.5, 5.98, 1.69 and 1.46, respectively. Average size of substitution segments also decreased with the number of times plants were backcrossed and selfed. In BC₂F₁, BC₃F₁, BC₃F₂ and BC₃F_3, average size of the segments was 25.43, 22.38, 20.78 and 18.15 cM, respectively. The rate of reduction of segment size was more in backcross (11.99%) than selfing (7.15%) generations. Percent recovery of recurrent parent genome in BC₂F₁, BC₃F₁, BC₃F₂ and BC₃F₃ was 82.24, 92.55, 98.04 and 98.52%, respectively. A total of 111 SSSLs comprising of 43 unique types were developed in BC₃F₂ and BC₃F₃. The estimated length of the segments in SSSLs ranged from 2.00 to 64.80 cM with an average of 21.75 cM, and 6.05 to 48.90 cM with an average of 20.95 cM in BC₃F₂ and BC₃F₃, respectively. Total length of all substitution segments was 2367.5 cM that covered 704.50 cM (39.25%) of the entire rice genome. Effective development and successful utilization of 3-S Lines for analysis of QTLs and mapping of genes established the suitability of the SSR marker facilitated backcross breeding approach for 3-S Lines development and its utilization.     

青海大黄油菜粒色性状分子标记的开发和图谱整合

利用青海大黄油菜和褐籽白菜型油菜09A-126构建BC₄和F₂分离群体, 结合AFLP与群体分离分析法(bulked segregant analysis, BSA)筛选引物, 获得5个与黄籽基因Brsc1紧密连锁的分子标记Y11~Y15。5个AFLP特异片段的序列, 均与白菜型油菜的A9染色体部分序列表现同源。将5个AFLP标记成功转化为5个SCAR标记(SC11~SC15)。利用目标基因所在染色体区段序列筛选到7个与目标基因紧密连锁的SSR标记(BrID10607、KS10760、B089L03-3和A1~A4)。利用SCAR和SSR标记扫描F₂群体中部分单株, 发现SC14和A1为共显性标记。用BC₄群体将Brsc1定位在标记Y06和A4之间1.7 Mb的区间内, 遗传距离分别为0.115 cM和0.98 cM。标记Y05和Y12与Brsc1共分离。本研究为黄籽油菜分子标记辅助选择育种体系的建立及目标基因的进一步精细定位和图位克隆奠定了基础。     

Genetic similarity among European winter triticale elite germplasms assessed with AFLP and comparisons with SSR and pedigree data

Genetic similarities (GS) based on molecular markers are well suited for direct exploration of relationships within a germplasm pool. The objectives of this study were to: (i) assess the genetic diversity in the European winter triticale germplasm by using AFLP markers, and (ii) compare the GS estimates of AFLP markers, simple sequence repeat (SSR) markers and MALÉCOT's coancestry coefficient (f). A representative set of 127 European winter triticale varieties and breeding lines, previously investigated with SSR, was assessed with 10 PstI/TaqI primer combinations (PC). AFLP analysis identified 344 polymorphic fragments with an average polymorphic information content per PC of 0.25 and a marker index of 8.56. GS‐values between genotypes (calculated after DICE) averaged 0.61 for AFLP and 0.43 for SSR. The mean f‐value was 0.06. Dendrograms based on ‘unweighted pair‐group method and arithmetic average’ showed no clear groupings within the triticale germplasm pool, but smaller clusters were consistently found. Both molecular marker systems were superior to the coancestry coefficient for genetic diversity assessment within the elite triticale germplasm.     

Association between AFLP-based genetic distance and hybrid performance in tropical maize

Identifying the best inbred combinations for the development of commercial hybrid maize varieties remains the main challenge to maize breeders. The aim of this work was to study associations between the genetic distance (GD) of 21 inbreds and the corresponding F₁ phenotypic data. Furthermore, the impact of grouping lines into genetically similar clusters was investigated. The 21 inbred lines were fingerprinted using amplified fragment length polymorphism markers. Parents and 210 F₁ progeny were evaluated in the field. Joint data analysis mostly revealed a tighter association between GD and the F₁ performance or mid parent heterosis in the intergroup than in the intragroup crosses. Despite these correlations, intergoup crosses should always be field‐tested before their release. Crosses showing low GD values should be discarded to avoid field‐testing costs. Better F₁ hybrid performance predictions can be achieved by integrating molecular and F₁ phenotypic data.     

Development of CGMS system in ridge gourd [<Emphasis Type="Italic">Luffa acutangula</Emphasis> (Roxb.) L.] for production of F<Subscript>1</Subscript> hybrids

Cytoplasmic male sterile system in ridge gourd has been converted to cytoplasmic genetic male sterile (CGMS) system through the development of analogues of male sterile (MS) line, maintainer line and fertility restorer line. These lines were developed by crossing the MS mutant, regenerated through in vitro culture, with monoecious pollen parents Deepthi, Haritham, LA 101, CO 2, IC 92761 and IC 92685. All hybrids and the BC₁ generation developed by crossing with the recurring pollen parents Deepthi, Haritham and LA 101 were male sterile. Male sterile BC₁ plants have been advanced to BC₆ generation and the parental line LA 101 was proved to be a successful maintainer line, producing male sterile progeny in successive back cross generations. Analogue of cytoplasmic male sterile line, MS LA 101, was developed through back crossing and on crossing with fertility restorer lines Arka Sumeet and LA 102, this line excelled as female parent, resulting heterotic combinations. Mitochondrial marker rpS14 and SCAR Tm-53 were identified to yield male sterility specific markers whereas SSR marker 18956 has generated the male fertility specific marker. These primers are recommended for marker assisted selection of ridge gourd, for utilizing male sterility for hybrid seed production and for developing A, B and C lines in CGMS system.     

小麦花粉致死基因Ki的SSR标记分析

小麦雄性不育主要是通过花粉的败育表现,其不育材料对小麦杂种优势的利用研究具有重要意义和价值,国外研究表明,某些特定普通小麦品种间杂交F₁表现的花粉部分不育现象,受控于核基因组花粉致死基因Ki,为了筛选小麦花粉致死基因Ki的连锁标记,利用现代分子生物学技术通过定位该基因,克隆出花粉致死基因连锁标记片段,为小麦雄性不育种质材料的转育提供有效的选择标记。对小麦花粉致死基因Ki进行了分子标记定位,以‘中国春’和澳大利亚春小麦品种的BC₁F₁代作为定位群体,利用分离群体分组分析法(BSA)对位于小麦6B染色体上85对SSR引物进行多态性筛选,具有多态性的引物再通过BC₁F₁定位群体进行验证,从中筛选出与目的基因连锁的2个SSR标记Xgwm626和Xgpw4138。运用Mapmaker 3.0软件进行连锁分析。结果表明,Xgwm626和Xgpw4138与Ki基因的遗传距离分别为9.2 cM和6.9 cM,且2个SSR标记位于目的基因两侧,并将Ki定位于小麦6BL染色体上。研究结果为Ki基因的分子标记辅助选择和进一步精细定位奠定了基础。     

Using yield quantitative trait locus targeted SSR markers to study the relationship between genetic distance and yield heterosis in upland cotton (Gossypium hirsutum)

The objective of this study was to examine the relationship between heterosis and genetic distance in upland cotton (Gossypium hirsutum L.). Heterosis in the different environments was evaluated and the relationship between heterosis and genetic distance (GD) was determined based on SSR markers for yield quantitative trait loci (QTL). Yields of seed cotton and lint showed a linear relationship with mid‐parent heterosis (MPH) and better‐parent heterosis (BPH). The variation in heterosis for other traits and their correlation with GD may be due to environmental factors or the effort of QTLs tested in the present study may vary in different environments. The present findings provide a foundation for heterotic grouping of parental lines and breeding of new cotton hybrids with improved seed cotton yield. This study calls for more research with stable QTLs as well as advance molecular marker techniques may be used in predicting yield heterosis in a more precise and reliable manner.     

利用2个F_2群体整合中国豌豆高密度SSR遗传连锁图谱

豌豆(Pisum sativum L.)是一种重要的食用豆类作物,在全世界范围内广泛种植,既可作为人类食物,也可作为牲畜饲料。用SSR标记构建的遗传连锁图谱在豌豆和其他作物的标记辅助育种中发挥着重要的作用。尽管对豌豆遗传连锁作图的研究已有悠久历史,但公众可获得且可转移的SSR标记以及基于遗传独特的中国豌豆种质的高密度遗传连锁图谱仍然有限。为了获得更多可转移的SSR标记和中国豌豆的高密度遗传连锁图谱,本研究首先从自主开发和文献获取的12,491个全基因组SSR标记中筛选了617个多态性SSR标记,并用于G0003973×G0005527 F_2群体遗传连锁图谱的加密。加密后的图谱全长扩展到5330.6 cM,包含603个SSR标记,标记平均间距离8.8 cM,相比之前的图谱有明显改善。基于上述结果,我们又筛选了119个具有多态性的SSR标记,用于构建大样本W6-22600×W6-15174 F_2群体的遗传连锁图谱,新图谱累积长度为1127.1 cM,包含118个SSR标记,装配在7条连锁群上。最后,将来自以上2个遗传图谱的数据进行整合,得到了一张覆盖范围6592.6 cM的整合图谱,包含668个SSR标记,由509个基因组SSR、134个EST-SSR和25个锚定标记组成,分布在7条连锁群上。这些SSR标记和遗传连锁图谱将为豌豆的遗传研究和标记辅助育种提供有力工具。     

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.     

Identification of a SCAR marker associated with Bm, the beet mosaic virus resistance gene, on chromosome 1 of sugar beet

Beet mosaic virus (BtMV) is an aphid transmitted, viral disease of beet found worldwide. The Bm gene, a resistance gene effective against BtMV, was identified in the sugar beet line 8500 and backcrossed into a C37 background to produce line C719. Three populations were developed from the cross of line C719 with the susceptible line C37 with the intent of developing markers for use in marker‐assisted selection. The F₂ progeny of three crosses were scored for resistance. Two of the three populations conformed to a 3 : 1 ratio, indicating a single gene trait. Sequence characterized amplified region (SCAR) markers were developed by using bulked segregant analysis combined with random amplified polymorphic DNA type markers. The markers showed close association to the Bm resistance gene and were effective in all three populations. The A₁ allele for genetic male sterility also was found to be associated with Bm and the SCAR marker. Development of a single‐nucleotide polymorphism marker from the SCAR sequence was used to validate linkage to chromosome 1 using separate mapping populations. This marker will be useful for the introgression of the Bm gene into germplasm.     

Mapping Co, a gene controlling the columnar phenotype of apple, with molecular markers

The columnar phenotype is a very valuable genetic resource for apple breeding because of its compact growth form determined by the dominant gene Co. Using bulked segregant analysis combined with several DNA molecular marker techniques to screen the F₁ progeny of Spur Fuji × Telamon (heterozygous for Co), 9 new DNA markers (6 RAPD, 1 AFLP and 2 SSRs) linked to the Co gene were identified. A total of 500 10-mer random primers, 56 pairs of selective AFLP primers and 8 SSR primer pairs were screened. One RAPD marker S1142₆₈₂, and the AFLP marker, E-ACT/M-CTA₃₄₆, were converted into SCAR markers designated SCAR₆₈₂ and SCAR₂₁₆, respectively. These markers will enable early selection in progenies where Co is difficult to identify. The Co gene was located between the SSR markers CH03d11 and COL on linkage group 10 of the apple genetic linkage map. Finally, a local genetic map of the region around the Co gene was constructed by linkage analysis of the nine new markers and three markers developed earlier.