Diversity and genetic differentiation among populations of Indian and Kenyan tea (Camellia sinensis (L.) O. Kuntze) revealed by AFLP markers

AFLP markers were successfully employed to detect diversity and genetic differentiation among Indian and Kenyan populations of tea (Camellia sinensis (L.) O. Kuntze). Shannon's index of diversity was used to partition the total phenotypic variation into between and within population components. On average, most of the diversity was detected within populations, with 79% of the variation being within and 21% being between populations of Indian and Kenyan tea. A dendrogram constructed on the basis of band sharing distinctly separated the three populations of tea into China type (sinensis), Assam type (assamica) and Cambod type (assamica ssp. lasiocalyx) in a manner consistent with the present taxonomy of tea, the known pedigree of some of the genotypes and their geographical origin. Principal coordinate (PCO) analysis grouped Assam genotypes both from India and Kenya supporting the suggestion that the Kenyan clones have been derived from collections made in this region. The China types were more dispersed on the PCO plot which is a reflection of wider genetic variation. As would be expected, clones collected from the same region exhibited less overall genetic variation. AFLP analysis discriminated all of the tested genotypes from India and Kenya, even those which cannot be distinguished on the basis of morphological and phenotypic traits. Received: 2 May 1996 / Accepted: 14 June 1996     

Genetic diversity of UPASI tea clones (Camellia sinensis (L.) O. Kuntze) on the basis of total catechins and their fractions

Tea leaf catechins and the ratio of dihydroxylated to trihydroxylated catechin fractions were analysed to identify the genetic diversity of 26 UPASI released tea clones. Principal component analysis (PCA) based on regression factor separated tea clones into five groups according to their jats (Jats are region based rays for e.g., Assam, China and Cambod origin) as well as their quality constituents (such as total polyphenols, total catechins, amino acids in the green leaves and liquor characteristics of black tea), particularly the catechins. Group 1 represented medium quality (quality of the final produce) clones, such as UPASI-10, UPASI-12 and UPASI-15 and drought tolerant clones like UPASI-1, UPASI-2, UPASI-9 and UPASI-10. Group 2 contained purely "China" cultivars while group 3 possessed high quality tea cultivars. "Assam" (group 5) teas had the lowest ratio of dihydroxylated to trihydroxylated catechin fractions (1:4) than the "Chinery" (group 2) teas (1:5). This biochemical differentiation indicated that there is a vast genetic diversity in UPASI released tea clones in terms of catechin fractions, even though the majority of them were selected from one tea estate located in the Nilgiris.     

Catechin and Catechin Fractions as Biochemical Markers to Study the Diversity of Indian Tea (Camellia sinensis (L.) O. Kuntze) Germplasm

The heterogeneous Indian tea germplasm includes ‘China’, ‘Assam’, ‘Cambod’, and their hybrids which were evaluated using biochemical markers viz., total catechin and their fractions, for varietal identification and characterization. Principal component analysis (PCA) of biochemical characters showed that the total catechin and trihydroxylated catechin has higher eigenvalues. The first two principal components (PCs) could differentiate more than 90% of the clones studied. This grouping based on first two principal component matrices differentiated ‘China’, and their hybrids with ‘Assam’ and ‘Cambod’ variety. Morphologically indistinct large‐leaved ‘Cambod’ variety and ‘Assam’ varieties could not be differentiated using biochemical markers, since both varietal types taxonomically belong to a single species. Clones of ‘China’ type showed low total catechin content and catechin ratio which are distinctly grouped. The ‘China–Assam’ and ‘China–Cambod’ hybrids formed intermediate groups between ‘China’ PC group and ‘Cambod’/‘Assam’ PC groups, providing evidence for genetic control of catechin ratio variation. Tea clones which are differentially positioned in the PC group could be explained based on the genetic contribution by other varietal type as parents. This biochemical characterization will be a useful tool in the development of quality‐tea clones with different proportion of total catechin and their fractions.     

大麦主栽品种亲缘系数和对叶斑病的抗性分析

为明确我国大麦主栽品种的遗传多样性及其对叶斑病的抗性来源,采用亲缘系数(COP,coefficient of parentage)分析方法对155个主栽大麦品种的遗传系谱进行聚类分析,同时对其中79个供试大麦品种在苗期和成株期分别接种2个强毒性菌株进行抗性鉴定。结果显示,155个品种聚为6个类群,有亲缘关系的品种占全部品种14.77%。在品种间组成的11935个组合中,1763个组合间存在亲缘关系,其COP值变化范围在0~0.7500之间,亲缘系数总和为157.5867,平均值为0.0132。根据系谱分析发现了不同育种单位所育品种的核心亲本,并追溯其主要的祖先亲本。此外,通过对叶斑病的抗性鉴定,发现大多数供试的大麦品种感叶斑病,高抗品种主要集中在垦啤麦系列品种和蒙啤麦3号,部分华大麦和驻大麦系列的品种在苗期或成株期中抗叶斑病。系谱分析及抗性鉴定结果揭示了我国大麦叶斑病抗性基因存在不同来源,分析结果有利于提高抗叶斑病基因筛选效率和缩小筛选范围,也将促进抗叶斑病新基因资源的发掘和利用。     

Pedigree‐based analysis of derivation of genome segments of an elite rice reveals key regions during its breeding

Analyses of genome variations with high‐throughput assays have improved our understanding of genetic basis of crop domestication and identified the selected genome regions, but little is known about that of modern breeding, which has limited the usefulness of massive elite cultivars in further breeding. Here we deploy pedigree‐based analysis of an elite rice, Huanghuazhan, to exploit key genome regions during its breeding. The cultivars in the pedigree were resequenced with 7.6× depth on average, and 2.1 million high‐quality single nucleotide polymorphisms (SNPs) were obtained. Tracing the derivation of genome blocks with pedigree and information on SNPs revealed the chromosomal recombination during breeding, which showed that 26.22% of Huanghuazhan genome are strictly conserved key regions. These major effect regions were further supported by a QTL mapping of 260 recombinant inbred lines derived from the cross of Huanghuazhan and a very dissimilar cultivar, Shuanggui 36, and by the genome profile of eight cultivars and 36 elite lines derived from Huanghuazhan. Hitting these regions with the cloned genes revealed they include numbers of key genes, which were then applied to demonstrate how Huanghuazhan were bred after 30 years of effort and to dissect the deficiency of artificial selection. We concluded the regions are helpful to the further breeding based on this pedigree and performing breeding by design. Our study provides genetic dissection of modern rice breeding and sheds new light on how to perform genomewide breeding by design.     

AFLP and pedigree-based genetic diversity estimates in modern cultivars of durum wheat [ Triticum turgidum L. subsp. durum (Desf.) Husn.

A substantial amount of between and within cultivar genetic variation was detected in all the 13 registered modern Canadian durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husn.) cultivars based upon amplified restriction fragment polymorphism (AFLP). Of the approximately 950 detected AFLP markers, only 89 were polymorphic, with 41 between cultivars whereas the remaining 48 showed polymorphism within at least one cultivar. The ancestry of Canadian durum wheat cultivars was traced back to 125 cultivars, selections, and breeding lines including 17 landraces. Mean pair-wise genetic distance based on the kinship coefficient was 0.76. On the other hand, AFLP-based mean pair-wise genetic distance was 0.40. Even though there was a large difference between the means of the two diversity measures, a moderate positive correlation (r=0.457, p<0.002) was detected between the two distance matrices. Cluster analysis with the entire AFLP data divided all cultivars into three major groups reflecting their breeding origins. One group contained ’Pelissier’ alone, which was a selection from a landrace introduced into the US from Algeria. On the other hand such groupings among cultivars were not evident when KIN was used for genetic diversity measures instead. The level of genetic variation among individuals within a cultivar at the breeders’ seed level was estimated based on an inter-haplotypic distance matrix derived from the AFLP data. We found that the level of genetic variation within the most-developed cultivars is fairly substantial despite rigorous selection pressure aimed at cultivar purity in breeding programs. Comparison of AFLP and pedigree-based genetic diversity estimates in crop species such as durum wheat can provide important information for plant improvement. Received: 26 January 2001 / Accepted: 31 May 2001     

Genetic diversity changes in Indian lentils over the times

The genetic diversity of 96 genotypes of lentil comprising 34 cultivars, 46 advanced breeding lines, and 16 germplasm lines were studied using 260 SSR markers. These markers generated a total of 749 alleles. The alleles/locus ranged from 2 to 16 with an average value of 2.87. Polymorphic information content varied from 0.02 to 0.91 with a mean of 0.30. Major allelic frequency ranged from 0.14 to 0.99 with a mean of 0.77. Studied genotypes were clustered into two groups according to their breeding history. Advanced breeding lines derived from exotic lines were clustered in one group, while another group accommodated most of the cultivars and advanced breeding lines with common cultivars in parentage. The germplasm lines were sub-clustered within first group. Cumulatively, first three principal components contributed 21.2% to the total variability. Advanced breeding lines showed higher number of alleles/locus and gene diversity (He) than other sets of genetic materials. In present study, no significant differences were observed between cultivars developed in different decadal groups for both NA and He. Moreover, genetic diversity changes between small and large seeded lentil cultivars were also found non-significant in this study. These findings showed that the use of alien genes can help to diversify active gene pool for developing improved new cultivars in lentil.     

Genetic diversity analysis of Bt cotton genotypes in Pakistan using simple sequence repeat markers

The popularity of genetically modified insect resistant (Bt) cotton has promoted large scale monocultures, which is thought to worsen the problem of crop genetic homogeneity. Information on genetic diversity among Bt cotton varieties is lacking. We evaluated genetic divergence among 19 Bt cotton genotypes using simple sequence repeat (SSR) markers. Thirty-seven of 104 surveyed primers were found informative. Fifty-two primers selected on the basis of reported intra-hirsutum polymorphism in a cotton marker database showed a high degree of polymorphism, 56% compared to 13% for randomly selected primers. A total of 177 loci were amplified, with an average of 1.57 loci per primer, generating 38 markers. The amplicons ranged in size from 98 to 256 bp. The genetic similarities among the 19 genotypes ranged from 0.902 to 0.982, with an average of 0.947, revealing a lack of diversity. Similarities among genotypes from public sector organizations were higher than genotypes developed by private companies. Hybrids were found to be more distant compared to commercial cultivars and advanced breeding lines. Cluster analysis grouped the 19 Bt cotton genotypes into three major clusters and two independent entries. Cultivars IR-3701, Ali Akbar-802 and advanced breeding line VH-259 grouped in subcluster B2, with very narrow genetic distances despite dissimilar parentage. We found a very high level of similarity among Pakistani-bred Bt cotton varieties, which means that genetically diverse recurrent parents should be included to enhance genetic diversity. The intra-hirsutum polymorphic SSRs were found to be highly informative for molecular genetic diversity studies in these cotton varieties.     

Genotyping of pedigreed apple breeding material with a genome-covering set of SSRs: trueness-to-type of cultivars and their parentages

Apple cultivars and breeding lines that represent much of the diversity currently present in major European breeding programmes and are genetically related by their pedigree were examined for the trueness of their identity and parentage by consistency in marker scores using a genome-covering set of 80 microsatellite (SSR) markers and an ??identity-by-descent?? approach. One hundred and twenty-five individuals were validated for the trueness-to-type of both their parents and 49 were validated for one of their parents, their second being unknown (23 individuals) or not available in this study (26 individuals). In addition, 15 individuals for which we lacked one of or both the direct parents were validated by consistency with tested parents of earlier generations. Furthermore, the identity of 28 founder cultivars was validated, their marker scores being consistent with descending cultivars and breeding lines. Four of the eight triploids identified were clearly shown to have arisen from unreduced egg cells. The assumed pedigree of 15 further individuals was found to be incorrect; fully consistent pedigrees were suggested for three of the cultivars. The pedigrees of a further eight individuals were confirmed through inference from the molecular data.     

The genetic base of Brazilian soybean cultivars: evolution over time and breeding implications

Genetic diversity is essential for crop breeding and one way to estimate it is through the concept of genetic base, which can be defined as the number of ancestors and their relative genetic contributions (RGC) to each cultivar. The RGC can be estimated through the coefficient of parentage between the ancestors and cultivars. Previous studies determined that the genetic base of Brazilian soybean was very narrow. The objective of this work was to evaluate the pedigree of 444 Brazilian soybean cultivars to estimate their genetic base. The cultivars were divided according to their release dates and according to their origin (public or private), and the genetic base for each group was also estimated. We found 60 ancestors, of which the top four (CNS, S-100, Roanoke and Tokyo, respectively) contribute 55.3% of the genetic base. Only 14 ancestors have an RGC over 1.0%, and they represent 92.4% of the genetic base. Analysis of the release dates indicated that there has been an increase in the number of ancestors over time, but the four main ancestors were the same over all periods, and their cumulative RGC increased from 46.6% to 57.6%, indicating a narrowing of the genetic base.     

SSR标记对不同黄籽甘蓝型油菜亲本材料的遗传分析

杂交育种中,亲本选配是育种成败的关键。本研究以重庆市油菜工程技术研究中心提供的180份甘蓝型黄子油菜亲本种质为材料,应用分布于不同连锁群的60对SSR标记进行了分析,共检测出308个标记位点,每对引物在不同亲本材料之间的等位基因数在1～11个之间,平均位点为5.1个。其中多态性位点207个,多态率达67.2%。对SSR扩增结果进行UPGMA分析,在遗传距离0.566处,180个品种(系)分为3个类群,聚类结果与种质来源比较一致,本研究为甘蓝型油菜黄子杂交育种和优势组合的选配提供了理论依据。     

Assessment of genetic variation within Indian mustard (Brassica juncea) germplasm using random amplified polymorphic DNA markers

Genetic diversity among 45 Indian mustard (Brassica Juncea L.) genotypes comprising 37 germplasm collections, five advance breeding lines and three improved cultivars was investigated at the DNA level using the random amplified polymorphic DNA (RAPD) technique. Fifteen primers used generated a total of 92 RAPD fragments, of which 81 (88%) were polymorphic. Of these, 13 were unique to accession 'Pak85559'. Each primer produced four to nine amplified products with an average of 6.13 bands per primer. Based on pairwise comparisons of RAPD amplification products, Nei and Li's similarity coefficients were calculated to evaluate the relationships among the accessions. Pairwise similarity indices were higher among the oilseed accessions and cultivars showing narrow ranges of 0.77-0.99. An unweighted pair-group method with arithmetic averages cluster analysis based on these genetic similarities placed most of the collections and oilseed cultivars close to each other, showing a low level of polymorphism between the accessions used. However, the clusters formed by oilseed collections and cultivars were comparatively distinct from that of advanced breeding lines. Genetically, all of the accessions were classified into a few major groups and a number of individual accessions. Advanced breeding lines were relatively divergent from the rest of the accessions and formed independent clusters. Clustering of the accessions did not show any pattern of association between the RAPD markers and the collection sites. A low level of genetic variability of oilseed mustard was attributed to the selection for similar traits and horticultural uses. Perhaps close parentage of these accessions further contributed towards their little diversity. The study demonstrated that RAPD is a simple and fast technique to compare the genetic relationship and pattern of variation among the gene pool of this crop.     

Extending the rapeseed gene pool with resynthesized <Emphasis Type="Italic">Brassica napus</Emphasis> II: Heterosis

Hybrid breeding relies on the combination of parents from two differing heterotic groups. However, the genetic diversity in adapted oilseed rape breeding material is rather limited. Therefore, the use of resynthesized Brassica napus as a distant gene pool was investigated. Hybrids were derived from crosses between 44 resynthesized lines with a diverse genetic background and two male sterile winter oilseed rape tester lines. The hybrids were evaluated together with their parents and check cultivars in 2 years and five locations in Germany. Yield, plant height, seed oil, and protein content were monitored, and genetic distances were estimated with molecular markers (127 polymorphic RFLP fragments). Resynthesized lines varied in yield between 40.9 dt/ha and 21.5 dt/ha, or between 85.1 and 44.6% of check cultivar yields. Relative to check cultivars, hybrids varied from 91.6 to 116.6% in yield and from 94.5 to 103.3% in seed oil content. Mid-parent heterosis varied from −3.5 to 47.2% for yield. The genetic distance of parental lines was not significantly correlated with heterosis or hybrid yield. Although resynthesized lines do not meet the elite rapeseed standards, they are a valuable source for hybrid breeding due to their large distance from present breeding material and their high heterosis when combined with European winter oilseed rape.     

Relevance of RAPD Analysis for Revealing Phylogenetic Relationships between Cultivars of Durum Wheat Triticum durum Desf.

A comparison of similarity indices between 64 durum wheat cultivars calculated using pedigree analysis and RAPD method showed a correspondence between these two approaches to estimation of genetic diversity. The associations between the results of RAPD clustering and coefficients of parentage (² test) and the coefficient of correlation between similarity matrices were statistically significant. However, the correlation was rather weak while pedigree analysis and RAPD method did not yield completely identical estimates of genetic diversity in the set of cultivars studied.     

Evaluation of the relevance of using RAPD-analysis for revealing the phylogenic connections between cultivars of drum wheat (T. durum Desf.)

A comparison of similarity indices between 64 durum wheat cultivars calculated using pedigree analysis and RAPD method showed a correspondence between these two approaches to estimation of genetic diversity. The associations between the results of RAPD clustering and coefficients of parentage (chi 2 test) and the coefficient of correlation between similarity matrices were statistically significant. However, the correlation was rather weak while pedigree analysis and RAPD method did not yield completely identical estimates of genetic diversity in the set of cultivars studied.     

AFLP-Based Genetic Diversity Assessment of Commercially Important Tea Germplasm in India

India has a large repository of important tea accessions and, therefore, plays a major role in improving production and quality of tea across the world. Using seven AFLP primer combinations, we analyzed 123 commercially important tea accessions representing major populations in India. The overall genetic similarity recorded was 51%. No significant differences were recorded in average genetic similarity among tea populations cultivated in various geographic regions (northwest 0.60, northeast and south both 0.59). UPGMA cluster analysis grouped the tea accessions according to geographic locations, with a bias toward China or Assam/Cambod types. Cluster analysis results were congruent with principal component analysis. Further, analysis of molecular variance detected a high level of genetic variation (85%) within and limited genetic variation (15%) among the populations, suggesting their origin from a similar genetic pool.     

Differential effect of the nitrogen form on the leaf gas exchange,amino acid composition,and antioxidant response of sweet pepper at elevated CO<Subscript>2</Subscript>

Cell wall invertases play an important role in plant growth and development, especially in the grain filling of crop plants. However, their potential in high yield crop breeding has not been investigated. In this study, the main hybrid maize cultivar Zheng Dan 958 (ZD958) was used as a basic variety to assess whether ZmGIF1, a cell wall invertase from maize, can be used to breed new cultivars with higher grain yields. ZmGIF1 expression, cell wall invertase activity, and sugar content in different parental inbred cultivars were compared with those in Zheng 58 and Chang 7-2, the parental inbred cultivars of ZD958. Parental cultivars which showed higher ZmGIF1 expression and invertase activity were selected and intercrossed to improve the expression of ZmGIF1. Compared with the basic cultivar ZD958, higher ZmGIF1 expression and cell wall invertase activity were observed in most hybrid F1 lines, leading to increased grain yield in them. All these results suggest that the expression of ZmGIF1 can be manipulated using different parental cultivars to increase the grain yield of their hybrid F1 progenies.     

我国茶树主要骨干亲本及其衍生品种(系)的SSR分析

铁观音、黄棪和福鼎大白茶分别是我国乌龙茶和红绿茶育种中的骨干亲本,由他们衍生出了一系列的优良品种,研究他们的遗传多样性及构建指纹图谱将有助于今后茶树育种工作中骨干亲本的合理利用和品种权的保护。本研究利用40对SSR引物对我国乌龙茶骨干亲本铁观音、黄棪及其衍生品种(系)和红绿茶骨干亲本福鼎大白茶及衍生品种进行了研究。结果表明,34份供试品种(系)的基因多样性指数（H）为0.54,平均遗传距离0.58,表明我国茶树主要骨干亲本及其衍生品种(系)具有较高的遗传多样性水平和较大的遗传变异,且90%的遗传多样性来自品种之间的遗传差异。聚类结果表明两套品种(系)各自聚为一类,遗传结构分析也显示两套品种(系)之间存在明显的差异。利用其中5对引物组合构建了供试材料的数码指纹图谱。     

Comparison of bread wheat lines selected by doubled haploid, single-seed descent and pedigree selection methods

 Yield performance of each group of ten spring bread wheat lines selected by doubled haploid (DH), single-seed descent (SSD) and pedigree selection (PS) methods from three F₁ crosses was compared with the aim of evaluating the DH method in breeding programs. Populations of 65–97 DH lines and 110 SSD lines per cross were used for selection. PS lines were developed by repeated selections from 1500 F₂ plants. Yield evaluation was performed at the F₆ generation of SSD and PS lines along with DH lines in a 2-year field experiment. It took only 2 years from the planting of wheat materials for DH production to the planting of selected DH lines for yield evaluation. There was no significant difference in grain yield between DH lines and PS lines selected from an F₁ cross whose parental varieties were closely related in their pedigrees. In two crosses with low coefficients of parentage and a large variation in their progenies, grain yield of selected DH lines was significantly lower than those of selected SSD and PS lines. These results confirm that the DH method can save time in obtaining recombinant inbred lines ready for yield evaluation. However, a larger DH population is required to achieve the same level of genetic advance with the PS method in crosses containing greater genetic variation. Received: 23 December 1997 / Accepted: 12 March 1998     

Analysis of genetic similarity detected by AFLP and coefficient of parentage among genotypes of sugar cane ( Saccharum spp.)

Despite the economical importance of sugar cane, until the present-date no studies have been carried out to determine the correlation of the molecular-based genetic similarity (GS) and the coefficient of parentage (f)-estimates generated for cultivars. A comprehensive knowledge of the amount of genetic diversity in parental cultivars, could improve the effectiveness of breeding programmes. In this study, amplified fragment length polymorphism (AFLP) and pedigree data were used to investigate the genetic relationship in a group of 79 cultivars (interspecific hybrids), used as parents in one of the Brazilian breeding programmes, and four species of Saccharum (Saccharum sinense, Saccharum barberi and two of Saccharum officinarum) . The objectives of this study were to assess the level of genetic similarity among the sugar-cane cultivars and to investigate the correlation between the AFLP-based GS and f, based on pedigree information. Twenty one primer combinations were used to obtain the AFLP molecular markers, generating a total of 2,331 bands, of which 1,121 were polymorphic, with a polymorphism rate, on average, of 50% per primer combination. GSs were determined using Jaccard’s similarity coefficient, and a final dendrogram was constructed using an unweighted pair-group method using arithmetic average (UPGMA). AFLP-based GS ranged from 0.28 to 0.89, with a mean of 0.47, whereas f ranged from 0 to 0.503, with a mean of 0.057. Cluster analysis using GS divided the genotypes into related subgroups suggesting that there is important genetic relationship among the cultivars. AFLP-based GS and f were significantly correlated (r = 0.42, P < 0.001), thus the significance of this r value suggests that the AFLP data may help to more-accurately quantify the degree of relationship among sugar-cane cultivars. Received: 27 November 2000 / Accepted: 27 April 2001