Development and applicability of GBS approach for genomic studies in Japanese plum (Prunus salicina Lindl.)

Genotyping by sequencing (GBS) provides a large quantity of useful data suitable for the identification of single nucleotide polymorphisms (SNPs), facilitating accurate genomic studies in plant species. In this study, GBS-based SNPs were used to characterise 11 Japanese plum cultivars and to explore their natural allelic diversity in relation to the most important phenology events (flowering date, ripening date and fruit development period) and fruit quality traits (weight, shape, skin and flesh colour, over colour, skin and flesh chlorophyll index, flesh firmness and soluble solids concentration). GBS-based SNPs were shown to be a powerful tool for genetic diversity and other genomic studies where SNP markers were related to several traits, particularly for flowering date, ripening date, fruit development period, skin chlorophyll degradation, flesh chlorophyll degradation and flesh colour. These results represent a preliminary approach using GBS as a possible breeding tool in current and new Japanese plum breeding programmes.     

Commercial Iranian olive cultivars: morphological traits,molecular diversity,and genetic structure

Iran is considered to have a unique gene pool of different fruit and nut species including olive (Olea europaea L.). In this study, we used 22 previously developed microsatellite (simple sequence repeat; SSR) markers for olive to evaluate the level of genetic variation and to produce identification keys for 63 Iranian accessions of olive belonging to 17 groups of cultivars. Based on morphological features, the number of flowers per inflorescence, fruit weights, endocarp weights, oil percentages, and flesh weights per endocarp had the highest coefficient of variation values, indicating the large extent of morphological variability among the 63 Iranian olive cultivars studied. All 22 microsatellite (SSR) markers revealed a high level of polymorphism, with a mean polymorphic information content (PIC) value of 0.511. Analyses of genetic structure among the 63 olive accessions were carried out using model-based methods, which showed a tendency for geographical clustering. Ten SSRs out of the 22 were successful for the identification of unique ID keys for 52 of the 63 accessions. In most cases, there was disagreement between the molecular data and the morphological data. These results could be used to reconstruct and maintain a collection of olive for future breeding programmes.     

我国建国以来苹果品种选育研究的回顾及今后育种的发展方向

培育具有自主知识产权的苹果品种对我国的苹果进入国际水果市场和抵御外来果品有着十分重要的意义。综合有关文献概述了我国建国以来在苹果育种方面取得的成就;分析了不同时期苹果育种的目标、采用的育种方法、培育的新品种及辅助育种技术的研究进展;并提出了苹果育种工作存在的问题和今后的发展方向。     

桃品种基因型分析及其利用

根据桃性状的遗传规律及表现，对桃育种材料的花粉育性、果实形状、果肉颜色、果肉肉质和核粘、离等质量性状进行基因型分析评价，同时讨论了白花、大久保、冈山３号、丰黄和连黄等优良种质在桃育种中的利用价值。     

Ascorbic acid level of Belgian apple genetic resources

With the help of the public, a wide collection of old apple cultivars (cvs) was established progressively at our Department from 1975 onwards. Currently, more than 1400 apple accessions are being maintained and three quarters of them are subjected to field evaluation for disease resistance and agronomic characters. This paper presents results of the ascorbic acid (Vitamin C) contents of old cvs combining interesting disease resistance and agronomic features in order to promote them as cvs adapted to production systems which are more environmentally friendly or to be used as parents in breeding programmes. First, a methodological study shows the importance of the variability of ascorbic acid content depending on fruit position in the trees, fruit size, fruit skin colour and maturity stage. Statistical analyses show that the sample size of six fruits per cv is sufficient to detect a difference of 20% between cvs. Old apple cvs such as Grenadier^RGF, Transparente de Croncels, Bramley’s Seedling, Reinette des Capucins, Reinette Libotte and Reinette de Waleffe contain three and seven times more ascorbic acid than the commercial cvs Gala and Elstar, respectively. The paper highlights the need to evaluate the rich potential in our genetic resources.     

Molekulargenetische Analyse des ‘Maschanzker’/‘Borsdorfer’‐Sortenkomplexes

‘Maschanzker’ is one of the best-known old apple cultivars in Austria and in the 19th century this variety also played an important role in the fruit production of South Tyrol (Northern Italy). ‘Maschanzker’ is believed to originate from one of the oldest German apple vari­eties ‘Borsdorfer’, which is even considered a synonym. However, since the denomination ‘Maschanzker’ is part of the name of a number of more or less similar types of apple, this study aimed at investigating the synonymy and homonymy as well as the relationships within and between different accessions of ‘Maschanzker’, ‘Borsdorfer’ and ‘Edelborsdorfer’ by employing 14 microsatellite DNA markers. A total of 38 accessions from several collections in Europe were analysed and 14 different genotypes were identified, which were assigned to three groups. Our data suggest a series of parent-offspring connections within the here defined Maschanzker group and the Edelborsdorfer group, while no closer relationship was evident to or among the accessions of the Borsdorfer group. Molecular genetic analyses thus provide fundamental data which can serve as a basis for the pomological revision of the ‘Maschanzker’/‘Borsdorfer’ cultivar complex.     

晚熟苹果新品种‘山农红’

‘山农红’苹果是从‘国光’芽变中选出的新品种。果实中等大小,扁圆形,平均单果质量182.9 g,大者可达260 g;果面光滑,底色黄绿,充分着色时全果面鲜红色,着色指数明显高于‘国光’;果肉黄白色,质地细密而脆,果汁中多,酸甜适口,含可溶性固形物14.5%,味浓,有芳香,品质上等;在山东牟平10 月中下旬成熟,果实发育期180 d 左右。盛果期产量118.8 t · hm-2,抗性与‘国光’相近。     

Genetic assessment of apple germplasm in Bosnia and Herzegovina using microsatellite and morphologic markers

Traditional apple cultivars from Bosnia and Herzegovina (B&H), potentially diverse due to specific geographic location and history of the country, represent a possible source of valuable traits for future breeding efforts and sustainable fruit growing. A total of 39 accessions, 24 traditional B&H cultivars and 15 modern international cultivars, maintained at the ex situ apple collection “Srebrenik” in Northeast Bosnia were, investigated using 10 SSR (simple sequence repeats) markers and 23 morphologic characteristics. All the used primer pairs manage to amplify clearly distinguishable and highly polymorphic SSR alleles, in average 10.4 alleles per locus. More than two different alleles per locus were detected for seven accessions (five traditional B&H cultivars and two international cultivars). Forty one unique alleles were exclusively present within the B&H cultivars, while seven unique alleles were only detected within international cultivars. The differentiation between traditional B&H and international cultivars (Fst = 0.060; P < 0.0001) was significant, also confirmed by analyses of molecular variance (AMOVA) (f_CT = 0.092; P < 0.001). Cluster analyses of 39 apple accessions, based on 10 SSR loci, revealed that only two traditional B&H cultivars grouped tightly with international cultivars (Ljepocvjetka and Bobovec Jon), while the rest formed separate clusters. Multivariate analyses of variance (MANOVA), nonparametric multivariate analyses of variance (NPMANOVA) and analyses of similarity (ANOSIM) showed statistically significant difference in morphologic characteristics between traditional B&H cultivars and the international cultivars. Cluster analyses of 39 apple accessions, based on the morphologic data, displayed less differentiation between traditional and international accessions, in comparison to the cluster analyses based on molecular data. No correlation between the molecular and morphologic data set was detected using the Mantel test. Many of the morphologic characteristics which have been analyzed in this study have significant commercial importance, we can assume that unlike the microsatellites these traits have been under agronomic selection pressure.     

栽培苹果起源研究进展

苹果是一种重要的温带落叶果树。研究栽培苹果的起源及其与苹果属植物野生种之间的进化关系,对未来的苹果育种及种质资源的保护和利用都具有重要的理论和实用价值。关于栽培苹果起源问题,现主要有2种观点:一种认为栽培苹果是由塞威士苹果直接栽培驯化而来,另一种观点认为塞威士苹果与其他苹果属植物野生种之间的杂交产生了现今的栽培苹果。综合有关文献,深入分析这2种观点,概述了近年来有关栽培苹果起源研究方面相关的实验证据,包括细胞学、孢粉学、分子标记以及DNA序列分析等方面的研究进展。提出了今后应当研究解决的关键问题。     

矿质元素含量与苹果风味品质及耐贮性的关系

对22个苹果品种果实矿质元素含量分析表明,钙、钾含量高,锰、铜含量低的品种果实肉质好,耐贮藏,具有良好的风味品质.钙、钾具有提高果实硬脆度、降低果实贮期的软化和失重、维持良好肉质和风味的作用.钙、钾含量高的品种果实硬脆度高,比重大,果肉致密,细胞间隙率低,贮期软化进度慢,肉质好,耐贮藏.锰、铜含量低的品种果实硬脆度高,含量高的品种果实韧性较强.锌含量对果实的风味、肉质和耐贮性影响较小,优质品种含锌量相对较低.     

Effects of position on canopy and harvest time on fruit physico-chemical and antioxidant properties in different apple cultivars

Fruit from Granny Smith, Fyriki, Fuji Kiku 8, and Imperial Double Red Delicious apple (Malus domestica Borkh.) trees planted in single or double rows, were harvested from different positions in the canopy, during the last month before commercial harvest. Fruit physico-chemical and antioxidant capacity, using the radical DPPH, were measured, in skin and flesh tissue. Greater total phenol content and total antioxidant capacity per fresh weight and serving portion of skin and flesh tissue, were found in Imperial Double Red Delicious and Fyriki apples, compared to Granny Smith and Fuji Kiku 8. The variation among cultivars in antioxidant contents was greater in peeled, compared with unpeeled fruit, highlighting the importance of eating unpeeled fruit. During the last two weeks before commercial harvest the total phenol content and total antioxidant capacity per fresh weight increased in skin of Granny Smith (by 24% and 42%, respectively) and Fuji Kiku 8 (by 19% and 27%, respectively). Fruit from the more sun—exposed parts of canopy had usually better red coloration and the effect was more pronounced in Fuji Kiku 8 and Fyriki, followed by Imperial Double Red Delicious and a lower effect was found in Granny Smith. Greater total soluble solid content, but not flesh firmness, were also found in fruit from the more sun—exposed parts of canopy, in all cultivars. The skin of fruit from the upper positions in the canopy had greater total phenol content and total antioxidant capacity, in all cultivars, apart from Fyriki. Antioxidant contents in flesh tissue were also greater in the upper positions of canopy in Fuji Kiku 8 and Imperial Double Red Delicious, and to our knowledge this is the first report on plant canopy effects on apple flesh antioxidant compounds.     

苹果果皮颜色性状的SSR标记

以24个苹果品种以及来自3个不同杂交组合的60个F1代结果单株为试材,对与苹果果皮颜色性状相关的DNA分子标记进行了研究。通过采用集群分类法(Bulked Segregant Analysis,BSA)对当前定位于苹果基因组图谱LG9上的所有SSR引物的分析,筛选到了一个与苹果果皮红色/非红色性状相关的SSR标记CN444542-156,该标记在红色表型上表现为隐性,在非红色表型上表现为显性。进一步分析表明,该标记对F1代单株性状预测的正确率为93.3%。     

Further Work on Plant Injection for Diagnostic and Curative Purposes

Summary

Randomly Amplified Polymorphic DNA (RAPD) markers were used to evaluate genetic similarity and inter-relationship among31 acid citrus species and cultivars, including sour oranges (six accessions); ‘Yuzu’ (four accessions) andits relatives (21 accessions). Out of the 60 decamer primers screened, 27 were selected which produced 108 markers; 76 of which were polymorphic. Species or cultivar-specific RAPD markers were also found. A dendrogram based on genetic distance implied that sour oranges were very distinct from ‘Yuzu’ and its relatives. ‘Yuzu’ accessions were very closely linked to each other, however; for the other specimens genetic polymorphism could easily be detected by RAPDs and the genetic variation between accessions was quite high and revealed their different origins. In this study some RAPDs allowed the distinction of very close cultivars, for instance ‘Kabosu’ from ‘Aka kabosu’.     

Genetic assessment of local apple cultivars from La Palma,Spain, using simple sequence repeats (SSRs)

Apple cultivars from Canary Islands can possibly be valuable genetic resources for subtropical areas. We localised 31 accessions considered by growers to be local, and confirmed by historical references that apple crop was introduced in XV century. These accessions were compared with 77 Spanish and 26 commercial cultivars in order to detect synonyms. A set of 10 SSRs were studied, and 2 of them presented 2 loci. Cultivars from La Palma (Canary Islands) presented five specific alleles not found in other Spanish regions. Those polymorphisms allowed detecting one introgressant in La Palma from non-native cultivars, and the other 30 accessions were classified into 14 genotypes. Some accessions derived from non-native cultivars such as Golden Delicious. A main cultivar could be detected, Del País, with 14 accessions. Secondary ones were Camuesa and Pero. Genetic differentiation was small between regions (Fst = 0.057) but significant, confirmed by analysis of molecular variance (AMOVA). Major genetic differentiation was found between non-native cultivars and cultivars from Asturias and Basque Country. Bayesian method and admixture analysis reconstructed three ancestral groups (RPP), Asturian and Basque cultivars grouped in RPPI (mainly those used for cider production), a mixture of cultivars from Galicia and La Palma in RPPII and non-native cultivars were in RPPIII. This genetic differentiation was also confirmed by factorial correspondence analyses (FCA). AMOVA over RPPs increased the genetic differentiation. Allelic variation found in this study showed that Spanish local cultivars represent a differentiated genetic pool that will provide original genotypes to diversify the reduced number of cultivars used in commercial production. In addition, differentiated genotypes localised in La Palma will be preserved in the local Germplasm Bank.     

Assessment of genetic diversity of Highland bananas from the National Banana Germplasm Collection at Rubona,Rwanda using RAPD markers

The genetic similarities of 49 accessions of bananas from The National Banana Collection at Rubona were investigated using random amplified polymorphic DNA markers. A total of 120 primers were screened for their usefulness in amplifying DNA fragments of four cultivars belonging to the subgroup Mutika–Lujugira. Fifteen random primers were selected for more detailed analysis, on the basis of providing reproducible amplification and revealing a high level of variation between the four cultivars. The genetic similarity was estimated using a simple matching coefficient which showed the lowest value of 0.46 between ‘Ingumba’ and ‘Ishika’ and the highest value of 0.85 between ‘Kirayenda’ and ‘Inyabukuwe’. The data of matrix of coefficient of similarity was subjected to cluster analysis using the unweighted pair group method with arithmetic average (UPGMA). Each accession was clearly separated. The results of this study are important for the curation of the banana germplasm collection in Eastern Central Africa and for future breeding of this crop.     

利用TP-M13-SSR标记构建苹果栽培品种的分子身份证

以国家果树种质兴城梨、苹果圃保存的314份来自19个国家的苹果栽培品种为试材,利用TP-M13-SSR标记,基于TP-M13-SSR指纹图谱构建苹果种质分子身份证。16对SSR引物在供试种质间共检测出等位基因357个,平均每对引物检测到22.3个。根据引物扩增等位基因数和Shannon’s指数,从1对引物开始逐步增加引物数量筛选可将供试材料全部区分的引物组合,最终确定6对核心引物可以区分全部供试苹果种质。基于供试苹果种质在6个SSR位点的指纹图谱,将等位基因编码成字符串获得分子身份证,利用条码技术其进一步转化成可被机器快速扫描的条码分子身份证,使每份种质具有可辨的分子身份证,达到利用最少、最特异引物区分最多苹果种质的目的。     

Association analysis of fruit traits in mulberry species (Morus L.)

Improvement of fruit traits is an important objective in current mulberry breeding programs. In this study, 93 mulberry accessions of diverse origin were genotyped using 15 ISSR markers to identify marker–trait associations with fruit traits. Fifteen ISSR primers generated a total of 104 amplification products, of which 94 were polymorphic, revealing 90.38% polymorphism; the mean PIC value was 0.2698. UPGMA cluster analysis showed clear genetic relationships between the 93 mulberry cultivars, and the major clusters were related to known pedigree relationships and their ecotype. The mean r² value for all intra-chromosomal loci pairs was 0.0210. Marker–trait associations were investigated using the unified mixed-model approach, considering both population structure (Q) and kinship (K). In total, 24 marker–trait associations (p < 0.01) were identified using different ISSR markers. The results suggest that association mapping in mulberry is a viable alternative to quantitative trait loci mapping, and detection of associations between markers and mulberry fruit traits will also provide important information for marker-assisted breeding.     

Evaluation of genetic diversity among Iranian soft-seed pomegranate accessions by fruit characteristics and RAPD markers

Soft-seedness in pomegranate is a desirable trait for fresh consumption of this valuable fruit. At the main Iran pomegranate collection, 21 pomegranate accessions gathered from different parts of Iran are registered as soft-seed genotypes. The aim of this research was to study these soft-seed pomegranate accessions using fruit morphopomological traits and DNA markers to reveal their relatedness. Thirty-six fruit characteristics were measured in these accessions together with applying 29 random decamer primers already reported to be polymorphic on pomegranate. Factor analysis on mean values of fruit characteristics determined 10 main factors and applied for grouping of the accessions using Ward's method. Also 14 of the random primers showed good amplification and polymorphism on these samples, and a total of 43 RAPD markers were produced. Estimates of genetic similarity, using Jaccard's similarity coefficient, ranged from 0.13 to 1.0 using the RAPD data. Grouping based on the fruit traits compared with that based on RAPD data did not produce a significant correlation (r = −0.36). Morphometric measurements and sensory evaluation confirmed that some accessions are hard or semi-hard seeded. This study showed that information based on fruit characteristics and RAPD markers are complementary for genetic discrimination in soft-seed pomegranate accessions. This might be due to the high level of similarity between soft-seed pomegranate accessions.