Nitrogen‐, water‐ and radiation‐use efficiencies affected by sugarcane breeding in Argentina

This study aimed to identify whether and how sugarcane (Saccharum spp.) breeding in Argentina modified nitrogen‐use efficiency (NUE), water‐use efficiency (WUE) and radiation‐use efficiency (RUE). Thirteen varieties were grown in two consecutive seasons. Trends in different traits were estimated by fitting the data to linear or bilinear regression models. There was a linear increase in NUE and WUE with the year of release throughout the 70‐year span, whereas water use was not modified by sugarcane breeding. There was a positive and strong (r > 0.90; P < 0.01) association between NUE and WUE and between sugar yield and NUE or WUE. Although RUE was not modified by sugarcane breeding, the amount of radiation intercepted by the crop increased with the year of release. Modern varieties had a higher maximum interception and needed fewer days to reach maximum interception than old varieties. This study suggests that applying ecophysiological knowledge would be instrumental in sugarcane breeding programmes in order to develop varieties with high resource‐use efficiency and capable to adapt to global climate change.     

Use of PCR-based methodologies for the determination of DNA diversity between Saccharum varieties

Summary In this study, two PCR-based methodologies were evaluated for potential use in the determination of DNA diversity between 20 commercial sugarcane hybrids and 6 outgroup varieties of S. spontaneum, S. officinarum and hybrids from early in the genealogy. The first method involved PCR amplification of sugarcane DNA in the presence of random, decamer primers (RAPDs), while the second protocol utilized specific microsatellite and telomere sequences as primers. A total of 41 RAPD primers (356 loci) were screened across the varieties of which 15 (160 loci) were used in the calculation of DNA diversity (expressed% similarity). This varied from 61 to 95%, with most of the commercial varieties showing more than 80% similarity in their DNA. The RAPD data indicated that there had been a gradual decline in DNA diversity (84% reduction) from the early inter-specific crosses to the commercial hybrids, probably as a result of backcrossing and in-breeding strategies used in the previous 5 to 6 generations of sugarcane breeding. The microsatellite and telomere data produced a much greater range in DNA similarity values (25–91%), probably due to the fact that these primers detect highly variable regions of the genome. It is suggested that these specific primers would not be suitable for determination of DNA diversity, but could be used more effectively in the development of a methodology for routine, rapid identification of sugarcane varieties.     

Selection of families and parents of sugarcane (Saccharum spp.) through mixed models by joint analysis of two harvests

At present, the sugarcane (Saccharum spp.) breeding programs from around the world have practiced early family selection, because in the early stages of breeding the evaluated characteristics have shown low heritability. This study aimed to select through restricted maximum likelihood/best linear unbiased prediction also known “mixed models” the best families among the 78 that comprise the RB05 sugarcane series, stage T1, of Programa de Melhoramento Genético da Cana-de-açúcar of the Rede Interuniversitária para o Desenvolvimento do Setor Sucroalcoleiro, Brazil. The experiments, conducted during 2007/2008 and 2008/2009 in Paranavaí, PR were implemented using an incomplete-block design with five replicates of each family. Each plot was composed of two planting rows of 5 m, spaced from each other by 1.40 m and spacing between plants was of 0.50 m, containing ten seedlings for planting row, resulting in a total of 20 individuals per replicate of each family. The characteristics evaluated were Brix (°Brix), ton of stalk per hectare (TCH) and ton of °Brix per hectare (TBH). The joint analysis obtained from these three traits in both harvests favored the selection of 35–41 families. The analysis also favored the selection of the best parents. The top five families for Brix were: F41M60, F02M77, F41M82, F61M38 and F62M35; to TCH were F66M30, F35M06, F78M45, F70M30 and F57M46 and for TBH the five elites families were F66M30, F35M06, F78M45, F70M30 and F01M39. The five best sugarcane parents for Brix were RB835486, SP91-1049, SP80-3280, RB72454 and RB925211; to TCH were IAC87-3396, RB941531, RB855511, RB915141 and RB957689, and for TBH the elite were IAC87-3396, RB855511, RB957689, RB941531 and RB855563. As each harvest season had different order of the best families, the joint analysis proved to be a primordial tool for plant breeders.     

引进国外马铃薯种质资源的遗传多样性分析

应用RAPD标记技术研究国外马铃薯品种的遗传多样性,以期明确这些品种的亲缘关系,为马铃薯优良品种的选育提供理论依据。利用筛选的29条RAPD引物,对24份国外马铃薯品种进行遗传多样性分析。结果显示,29条RAPD引物共扩增出165个条带,其中116条为多态性条带,多态性比例为70.3%;试供的24个国外马铃薯品种间的遗传相似系数在0.514~0.816之间,平均为0.677;相似系数小于0.6的品种对仅为7.25%;24个马铃薯品种可聚为2大类6亚类。结果表明,试供的24个国外马铃薯品种间的遗传相似性高,遗传基础比较狭窄。     

Genetic diversity in cultivated Osteospermum as revealed by random amplified polymorphic DNA analysis

A collection of 28 Osteospermum clones and cultivated varieties of different origin were evaluated by random amplified polymorphic DNA (RAPD) analysis. All the clones were identified by 12 decamers selected from a set of 30. This is the first characterization by molecular markers of the genetic material of Osteospermum. The level of genetic diversity among genotypes was assayed and all the accessions tested were then classified into six groups by UPGMA cluster analysis; the clustering of genotypes using the RAPD data proved to be in accordance with their breeding group origin. RAPD analysis can therefore be a useful tool for evaluating genetic variability in other Osteospermum germplasm collections for breeding purposes and for protecting intellectual property rights of improved varieties.     

Molecular diversity and genome structure in modern sugarcane varieties

Summary RFLP analysis was performed on 40 sugarcane cultivated varieties. Twenty-two maize low copy DNA clones located on different regions of the 10 maize chromosomes were used as probes to survey variability among the sugarcane varieties. A total of 425 fragments, 411 of which were polymorphic, were identified for 22 probe/enzyme combinations. Each variety displayed an average of 7.28 fragments per combination, revealing the complex polyploid origin of modern sugarcane varieties. The average genetic similarity between sugarcane varieties was 0.61. Although cultivated varieties appear closely related to S. officinarum clones, the genes of S. spontaneum seem to constitute the principal component of varietal diversity. A very weak global structuring among the 40 varieties is observed, in agreement with the profuse exchanges of parental materials between sugarcane breeding stations. Traces of linkage disequilibrium can be attributed to the distribution of S. spontaneum chromosomes among sugarcane varieties. The possibility of using modern varieties as a population for detecting associations between molecular markers and agronomic traits is suggested.     

Identification of RAPD markers closely linked to the mlo-locus in barley

Developing resistance to powdery mildew, Erysiphe graminis f.sp. hordei, is a major goal of many barley breeding programmes. Several resistance genes have been tagged or mapped with molecular markers. The mlo gene confers durable resistance towards all known isolates of the pathogen. In this study, RAPD markers and bulked segregant analysis were used to determine PCR-based markers linked to the mlo-locus. Sixty doubled haploid lines from a cross between an isogenic line of ‘Ingrid’ carrying the mlo11 allele and a susceptible cv. ‘Pokko’ were used as plant material. Seven linked RAPD markers were found, the closest lying 1.6 cM away from the resistance gene. When eight barley varieties were assayed for the presence of this band, F4-980, it was found in the resistant varieties but not in the susceptible ones. The linked marker bands could be amplified from DNA-samples prepared by using three different methods, including a quick squash technique. PCR-based markers linked to the resistance gene can be used as tools for selection in breeding programmes.     

Application of RAPD markers in the characterisation of Chrysanthemum varieties and the assessment of somaclonal variation

Characterisation of fifteen commercial varieties of Chrysanthemum was carried out through RAPD analysis. Varieties could be distinguished from each other and the level of similarity between varieties seemed to be not very high. In vitro cultures were establish from four varieties and were subjected to different proliferation conditions. Five individuals from each variety and treatment were analysed using RAPD at the beginning of the treatment and after a month of culture. Variation was detected at both stages of the culture period. The rate of variation found showed differences between varieties, but no significant difference was found between culture conditions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic diversity in soybean genotypes from north‐eastern China and identification of candidate markers associated with maturity rating

Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to estimate the genetic relationships among 101 soybean cultivars developed in north‐eastern China. Fifty‐three fragments of the 100 RAPD markers and 35 SSR markers tested were polymorphic across the 101 soybean cultivars. Similarity values among these soybean cultivars ranged from 45.2% to 100% for RAPD data, and ranged from 36.1% to 100% for SSR data. The similarity matrices for SSR data and RAPD data were moderately correlated (r = 0.31, P < 0.05). Cluster analyses indicated that the cultivars released from the same seed company were mostly grouped together. A principal component analysis, based on the combined RAPD and SSR data, yielded a good separation of soybean varieties with different maturity ratings [represented by soybean Heat Unit (HU)]. The varieties with HU < 2200 were well separated from those with HU > 2200. Four RAPD markers and eight SSR markers were significantly associated with the maturity ratings of soybean.     

SCAR and RAPD markers associated with 18-carbon fatty acids in rapeseed, Brassica napus

Breeding rapeseed for enhanced oil quality includes the development of varieties with low linolenic acid content. The breeder also aims to develop varieties with a high linoleic acid content because of its nutritional value. Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers have been developed for linolenic acid content, but they are not best suited for a direct application in marker-assisted selection. The RFLP technique is too complex and time-consuming and RAPD markers lack codominance, precluding the distinction of homozygous from heterozygous individuals. In this report the conversion of a RAPD marker to a codominant sequence characterized amplified region (SCAR) marker named L1L9 is described. One of the alleles consisting of an 899 bp fragment (allele A), is associated with low linolenic acid content. The other allele consists of an 641 bp fragment (allele B) and is associated with high linolenic acid content. This marker explains approximately 25% of the genetic variation for this trait. Linkage analysis in the mapping population indicates that the SCAR marker probably tags an ω-3 desaturase gene in B. napus. Two RAPD markers were found to be associated with oleic/linoleic acid content. Markers M14-350 and I06-650 explained approximately 10% and 7% of the genetic variation for linoleic acid content, respectively. These two markers were found linked at 12.3cM in the segregating B. napus F₂ progeny used for mapping. All the markers reported in this paper should be useful in breeding programmes for developing high linoleic and low linolenic acid rapeseed varieties.     

Spanish spelt: a separate gene pool within the spelt germplasm

The renewed interest in spelt (Triticum spelta L.) for wheat improvement programmes requires the study of the available genetic diversity. The purpose of this study was to assess genetic diversity within a Spanish spelt collection. Sixty‐six Spanish spelt accessions, 19 accessions of T. spelta and T. macha from different origins, three bread wheat cultivars (T. aestivum) and one accession of T. dicoccum were screened using simple sequence repeats (SSRs). The diversity observed within the Spanish group was comparable with that observed in the other wheat varieties, despite their broader geographical diversity. Indeed, the highest polymorphic information content value calculated with SSRs for Spanish material (0.90) is similar to that observed for the other wheat varieties (0.98). Principal component analysis explained 46.5% of the cumulative variation and confirmed the Spanish accessions as a separate group. This study showed the Spanish spelt collection to be a variable and unique genetic resource for wheat and spelt breeding programmes.     

不同基因型甘蔗甘露糖抗性筛选体系的建立

为明确不同基因型甘蔗再生过程中对甘露糖的敏感性,为甘蔗基因工程育种中转化子的离体筛选提供技术。使用甘露糖作为筛选底物,对甘蔗品种‘ROC22’、‘桂糖94-119’和‘桂糖96-44’外植体的再生进行研究。建立3个基因型甘蔗的甘露糖抗性筛选体系,在甘蔗愈伤组织继代增殖、分化和小苗生根3个阶段进行抗性筛选时,甘露糖占总糖的比例分别是60%、60%~70%和30%。通过统计分析,明确了不同再生阶段甘露糖对3个甘蔗基因型的影响不同,继代增殖阶段基因型间无明显差异,但在分化和生根阶段达极显著差异。     

Molecular mapping of black rot resistance locus Xca1bo on chromosome 3 in Indian cauliflower (Brassica oleracea var. botrytis L.)

Black rot is the most devastating disease of cauliflower worldwide causing severe damage to crop. The identification of markers linked to loci that control resistance can facilitate selection of plants for breeding programmes. In the present investigation, F₂ population derived from a cross between ‘Pusa Himjyoti’, a susceptible genotype, and ‘BR‐161’, a resistant genotype, was phenotyped by artificial inoculation using Xcc race 1. Segregation analysis of F₂ progeny indicated that a single dominant locus governed resistance to Xcc race 1 in ‘BR‐161’. Bulk segregant analysis in resistant and susceptible bulks of F₂ progeny revealed seven differentiating polymorphic markers (three RAPD, two ISSR and two SSR) of 102 markers screened. Subsequently, these markers were used to genotype the entire F₂ population, and a genetic linkage map covering 74.7 cM distance was developed. The major locus Xca1bo was mapped in 1.6‐cM interval flanked by the markers RAPD 04₈₃₃ and ISSR 11₆₃₅. The Xca1bo locus was located on chromosome 3. The linked markers will be useful for marker‐assisted resistance breeding in cauliflower.     

Application of in vivo and in vitro mutation techniques for crop improvement

Summary Conventional mutation techniques have often been used to improve yield, quality, disease and pest resistance in crops, or to increase the attractiveness of flowers and ornamental plants. More than 1700 mutant varieties involving 154 plant species have been officially released. In some economically important crops, e.g. barley, durum wheat and cotton, mutant varieties occupy the majority of cultivated areas in many countries. Mutation techniques have become one of the major tools in the breeding of ornamentals such as alstroemeria, begonia, chrysanthemum, carnation, dahlia and streptocarpus. The use of in vitro techniques such as anther culture, shoot organogenesis, somatic embryogenesis and protoplast fusion can overcome some of the limitations in the application of mutation techniques in both seed and vegetatively propagated crops. In vitro culture in combination with induced mutations can speed up breeding programmes, from the generation of variability, through selection, to multiplication of the desired genotypes. The expression of induced mutations in the pure homozygote obtained through microspore, anther or ovary culture, can enhance the rapid recovery of the desired traits. In some vegetatively propagated species, mutations in combination with in vitro culture technique, may be the only method of improving an existing cultivar. Currently, many molecular studies rely on the induction and identification of mutants in model species for construction and subsequent saturation of genetic maps, understanding of developmental genetics and elucidation of biochemical pathways. Once identified and isolated, the genes that encode agronomically-important features can be either introduced directly into crop plants or used as probes to search for similar genes in crop species. It seems most likely that the recent developments based on these technologies will soon provide improved methods for selection of desired mutants.     

Independent target region amplification polymorphism and single‐nucleotide polymorphism marker utility in genetic evaluation of sugarcane genotypes

The independent target region amplification polymorphism (TRAP) and single‐nucleotide polymorphism (SNP) marker s were used for genetic evaluation of different selected 47 sugarcane genotypes. A total of 23 pairs of TRAP markers generated 925 alleles, of which 74% alleles were polymorphic. Polymorphism was generally high (>50%), ranging from 54 to 98%. The polymorphism information content (PIC) values 0.20 varied among the primer combination ranging from 0.17 in SAI + Arbi 2 to 0.31 in GL 2+ Arbi 1 with an average of 0.24. However, the Pearson correlation between PIC and power of discrimination (PD) was found to be less significant. Single‐nucleotide polymorphisms were used first time for the assessment of genetic diversity among different species of Saccharum and cultivated sugarcane varieties. The SNPs were detected from 454 sequencing. A total of 245 SNP markers were assayed across the 47 genotypes, and 167 SNPs were found to be polymorphic. The PIC values ranged from 0.04 to 0.38 with an average of 0.21, and their respective PD varied from 0.58 to 0.04 with an average value of 0.31. The obtained results relatively significant were compared with the other marker systems through genetic similarity and the clusters formed in different unweighted pair group method with arithmetic mean clustering dendrogram. The clustering analysis established genetic relationship in the order of Erianthus > Sclerostachya > Narenga > Saccharum spontaneum > S. robustum > S. barberi > S. officinarum/cultivars. These results ratify TRAP and SNP marker systems for assessing genetic diversity studies, and more diversified Erianthus spp. can contribute substantially towards sugarcane varietal improvement through breeding with Saccharum spp. or hybrid cultivars.     

甘蔗品种的AFLP和SSR标记鉴定及其应用

品种遗传多样性和指纹图谱是育种、品种权保护和新品种推广等工作的重要参考和依据。本研究选用38个来自国家甘蔗品种区域试验的甘蔗新品种(系), 以9对AFLP标记扩增出348个位点, 多态性位点248个, 多态性比率为72.26%; 15对SSR标记扩增出180个位点, 多态性位点176个, 多态性比率为97.78%。38个新品种(系)的遗传相似系数分布在0.668~0.847之间, 其箱线图分布特征显示, 其中的FN、MT、YZ、YG、GT等系列品种(系)遗传基础相似。通过UPGMA聚类表明, 可在遗传相似系数为0.732处将38个甘蔗新品种(系)划分为2个群体, 其中福农09-2201和桂糖06-1492作为一个子群体最先被划分出来, 它们在群体中的异质性较强; 另外, 在遗传相似性系数为0.770处划分出一个子群体a, 其中包含参照品种ROC22、福农07-3206、福农40、海蔗22、桂糖09-12、柳城07-150等品种(系)。ROC22具有广适应性和高产高糖等优良特性, 子群体a中的另外几个品种(系)则更有可能拥有这些特性, 具有更高的推广潜力。本研究选择60个SSR位点构建了38个甘蔗新品种(系)的指纹图谱, 对品种鉴定及品种权的保护具有重要作用, 可望直接应用于指导甘蔗种质资源的遗传多样性评价和分子指纹图谱鉴定, 并将为这些品种(系)推广布局或作为杂交亲本利用提供参考和借鉴。     

Assessment of natural and induced genetic variation in Alstroemeria using random amplified polymorphic DNA (RAPD) markers

Summary We have used random amplified polymorphic DNA (RAPD) markers to study genetic variation in Alstroemeria. The first objective was to examine the discriminatory power of RAPD markers in different genotypes of Alstroemeria obtained by traditional breeding. All genotypes examined, including commercial Alstroemeria varieties, could be distinguished on the basis of their RAPD profiles. Progeny plants could be distinguished from their parents. A second objective of this study was to investigate whether RAPD markers can be used as a routine tool to detect mutant plants, as an alternative to glasshouse testing. To address this objective, we analysed Alstroemeria plants that carried phenotypically visible mutations that either were induced by irradiation using X-rays or were the result of somaclonal variation. In eight out of a total of 13 mutant Alstroemeria plants obtained after irradiation or tissue culture we detected no polymorphisms when compared to control plants that were considered to be non-mutated. Only in five of the mutant plants analysed we detected one to two polymorphisms. These results suggest that frequent genome rearrangements had not occurred in the mutant plants analysed. These results also demonstrate that the RAPD technique is an inappropriate tool for the rapid screening of Alstroemeria for induced variation. It that the RAPD technique is an inappropriate tool for the rapid screening of Alstroemeria for induced variation. It seems probable that this conclusion would be equally applicable in other plant genera in which induced variation has occurred. However, the RAPD technique is a simple and effective tool for genetic fingerprinting of Alstroemeria varieties, provided their differences are due to sexual propagation.     

Genetic relationships and hybrid vigour in olive (Olea europaea L.) by microsatellites

The olive (Olea europaea) is one of the most important oleaginous crops of the Mediterranean basin. Increased demand for olive oil creates a need for new olive varieties to help meet the requirements of the global market. However, olive breeding has been handicapped by such varied challenges as a prolonged juvenile period, agrotechnical problems and insufficient genetic knowledge. The use of DNA markers has the potential to overcome these problems and increase the effectiveness of classical breeding programmes. In this study, co‐dominant polymorphic simple sequence repeats (SSRs) were used as markers to analyse the genetic relationships between several local and other ‘non‐native’ olive cultivars. Cluster analysis revealed four major groups among the 15 cultivars examined in this study. Table and oil cultivars were clustered in different groups. However, the clusters did not differentiate between cultivars of different geographical origins. In addition, we used the data gathered to analyse genetic relationships to evaluate the effects of heterosis in agricultural traits. Genetic distances between cultivars were determined based on the SSR genotype data and were used for evaluating the possible effects of heterosis in various F₁ populations. Interestingly, phenotypic data of F₁ progenies from crosses between different cultivars indicated the potential effects of heterosis as expressed in several traits. Genetic distance between parents was significantly correlated to F₁ performance for three traits: percentage of dry fruit weight, oil content and commercial oil production. Thus, crosses between olive cultivars exhibiting relatively extensive genetic distances one from the other are expected to result in better progeny performance in future Olea breeding programmes. Our study linked assessment of biodiversity of commercial olive cultivars with the application of this information in olive breeding programmes for selection of specific parents to generate superior new cultivars.     

Genetic variability measures among Bromus catharticus Vahl. populations and cultivars with RAPD and AFLP markers

The objectives of this study were to optimize RAPD and AFLP techniques in B. catharticus, and to determine the genetic variability of populations and commercial prairie grass cultivars with the aforementioned molecular markers. Two populations with contrasting morphological characteristics were evaluated from individual and bulked DNA samples using RAPD markers. Both analyses showed a similar information about inter population variability. Each accession was sampled by a single leaf bulk of 10 plants. Accession similarities were established with 276 RAPD and 714 AFLP bands using Jaccard similarity coefficient. The dendrogram of the accessions using RAPD markers showed that they shared high similarity values (>94%). A similar result was obtained with AFLP markers (similarity values >98%), revealing a narrow genetic basis in the analyzed accessions. Consequently, molecular characterization of germplasm should be considered in addition to morphological criteria, to choose the parental genotypes for breeding programs of this forage crop. The AFLP technique was more efficient to detect DNA polymorphism in our experiments and unique fingerprints were detected for all the accessions. RAPD is a simple and non expensive technique, suitable to estimate genetic similarity. This revised version was published online in August 2006 with corrections to the Cover Date.     

Feeding quality assessment of fresh maize stover by means of near-infrared spectroscopy with a new sample presentation unit

Stover quality traits are important in breeding programmes of maize cultivars. However, conventional procedures for evaluation of stover quality are expensive and thus limit the full exploitation of the available genetic variability. Our objective was to assess the potential of near‐infrared spectroscopy (NIRS) with a new sample presentation unit to determine feeding quality of maize stover. The plant material comprised maize inbreds and hybrids. The sample presentation unit was equipped with a near‐infrared diode‐array spectrometer. Coefficient of determination in cross‐validation ranged between 0.77 and 0.94 for contents of dry matter, nitrogen, neutral detergent fibre and in vitro fermentability expressed as gas volume after 24h incubation time, but it was lower for ash content. It was concluded that NIRS with the new sample presentation unit was less accurate than laboratory NIRS to predict the standard reference methods. However, the new sample presentation unit might be used as a fast and efficient technique to perform large screenings of maize stover accessions in breeding programmes.