Comparative analysis of DNA polymorphisms and phylogenetic relationships among Syzygium cumini Skeels based on phenotypic characters and RAPD technique

The Indian black berry (Syzygium cumini Skeels) has a great nutraceutical and medicinal properties. As in other fruit crops, the fruit characteristics are important attributes for differentiation were also determined for different accessions of S. cumini. The fruit weight, length, breadth, length: breadth ratio, pulp weight, pulp content, seed weight and pulp: seed ratio significantly varied in different accessions. Molecular characterization was carried out using PCR based RAPD technique. Out of 80 RAPD primers, only 18 primers produced stable polymorphisms that were used to examine the phylogenetic relationship. A sum of 207 loci were generated out of which 201 loci found polymorphic. The average genetic dissimilarity was 97 per cent among jamun accessions. The phylogenetic relationship was also determined by principal coordinates analysis (PCoA) that explained 46.95 per cent cumulative variance. The two-dimensional PCoA analysis showed grouping of the different accessions that were plotted into four sub-plots, representing clustering of accessions. The UPGMA (r = 0.967) and NJ (r = 0.987) dendrogram constructed based on the dissimilarity matrix revealed a good degree of fit with the cophenetic correlation value. The dendrogram grouped the accessions into three main clusters according to their eco-geographical regions which given useful insight into their phylogenetic relationships.     

A short history of Lagenaria siceraria (bottle gourd) in the Roman provinces: morphotypes and archaeogenetics

Lagenaria siceraria (Molina) Standl. (bottle gourd) is pantropic and displays large variation in fruit and seed shape. Two subspecies are currently recognized: the African L. siceraria ssp. siceraria and the Asian L. siceraria ssp. asiatica. The Asian type of bottle gourd belongs to the earliest domesticated plants in the Americas. In Europe, bottle gourd only appears with some frequency from the Roman period onwards. The paper is the study of ancient DNA (aDNA) and seed morphology of one almost complete bottle gourd fruit from the Roman site of Oedenburg/Biesheim–Kunheim, France (1st century a.d.), and from individual seed finds from the Roman vicus of Petinesca–Vorderberg, Switzerland (3rd century a.d.), both recovered from waterlogged layers. Width and length measurements of seeds show large variation. Based on the index of width to length, seeds from both sites differ significantly (p?<?0.0001 Mann–Whitney) suggesting that there were different variants present north of the Alps. Genetically, the bottle gourd fruit from Roman Oedenburg/Biesheim–Kunheim and one commercial cultivar L. siceraria cv. ‘Herkuleskeule’ are of Asian origin as identified by three Asian and African specific chloroplast markers. These results support an early and long-lasting presence of the Asian type of domestic bottle gourd in Europe. No chloroplast markers were found in the seeds from Petinesca–Vorderberg. However preserved nuclear high copy 5.8S rDNA fragments correctly matched to Cucurbitaceae, further supporting the evidence for preservation of DNA in waterlogged plant remains.     

中国特有植物艾比湖沙拐枣(Calligonum ebi-nuricum)居群内果实性状的变异

以中国荒漠特有植物艾比湖沙拐枣(Calligonum ebi-nuricum)果实形态特征为研究对象,对分布于原产区的同一居群12个样株的果实进行形态特征比较研究。通过每株30粒果实的喙长、果实长度、果实宽度、果实形状、瘦果长度、瘦果宽度、瘦果形状、刺毛长度、头部刺毛长度、两肋间距、两刺间距11个性状指标的数值分析表明：在株内与株间均有变异,株间果实性状变异较株内果实性状变异大;代表果实和瘦果大小指标正相关性极显著（p＜0.01）;两肋间距与瘦果形状呈负相关（r=-0.297**,p＜0.01）。研究结果证明：其果实形状和瘦果形状（长度、宽度及长宽比）较为稳定,可作为艾比湖沙拐枣的分种依据,而其它果实性状可塑性较大,只能作为分类的参考。该研究结果可供沙拐枣属其它种的准确分类参考。     

Comparison of SSR and cytochrome P-450 markers for estimating genetic diversity in Picrorhiza kurrooa L.

Picrorhiza kurrooa L., a high altitude medicinal plant, is known for its drug content called Kutkin. In the present study, DNA-based molecular marker techniques, viz. simple sequence repeats (SSR) and cytochrome P-450 markers were used to estimate genetic diversity in Picrorhiza kurrooa. Twenty five accessions of Picrorhiza kurrooa, collected from ten different eco-geographical locations were subjected to 22 SSR and eight cytochrome P-450 primer pairs, out of which 13 SSR markers detected mean 5.037 alleles with a mean polymorphic information content (PIC) of 0.7718, whereas eight cytochrome P-450 markers detected mean 5.0 alleles with a mean PIC of 0.7596. Genetic relationship among the accessions was estimated by constructing the dendrograms using SSR and cytochrome P-450 data. There was a clear consistency between SSR and cytochrome P-450 trees in terms of positioning of most Picrorhiza accessions. SSR markers could cluster various Picrorhiza kurrooa accessions based on their geographical locations whereas cytochrome P-450 markers could cluster few accessions as per their geographical locations. The Mantel test between SSR and cytochrome P-450 markers revealed a good fit correlation (r = 0.6405). The dendrogram constructed using the combined data of SSR and cytochrome P-450s depicted two clusters of accessions based on its eco-geographical locations whereas two clusters contained the accessions from mixed eco-geographical locations. Overall, the results of the present study point towards quiet high degree of genetic variation among the accessions of each eco-geographic region.     

基于形态及分子标记的濒危植物夏蜡梅自然居群的遗传变异研究

比较了濒危植物夏蜡梅(Sinocalycanthus chinensis)大明山、大雷山、龙须山3个居群的果实与种子的形态变异,采用ISSR分子标记技术分析DNA序列的变异,综合评价夏蜡梅3个居群的遗传变异。结果如下:果实性状中,果柄长、果实长、果实重、每果种子数等指标均以大明山居群最大、大雷山居群次之、龙须山居群最小,它们之间差异显著(P<0.05)。种子重、种子长、种子宽、种子厚等指标均以大雷山居群最大,与大明山居群和龙须山居群差异显著(P<0.05)。基于果实形态特征的表型分化系数(VST)在0.5518—0.9750之间,平均为0.8930,所有果实形态指标的变异大部分存在于居群间。基于种子形态特征的VST在0.1669—0.8678之间,平均为0.6240,除种子长外,其他种子形态指标的变异也大部分存在于居群间。ISSR分析表明,3个居群的多态位点百分率、Shannon信息指数、Nei基因多样性均是大明山居群最高、龙须山居群次之、大雷山居群最低。居群间的遗传分化系数(GST)为0.6050。基于ISSR分子标记数据的聚类结果显示大明山居群先与龙须山居群聚在一起,再与大雷山居群相聚,这与基于种子性状特征的聚类结果相似,而与基于果实性状特征的聚类结果不同。3个居群的种子形态特征变异系数与遗传多样性水平具有显著的正相关,且基于果实形态特征估算的VST要高于GST,而基于种子形态特征估算的VST仅略高于GST,表明夏蜡梅不同居群间的种子形态变异主要由遗传变异所造成的,而环境因子在果实形态变异中起了重要作用。Mantel检验显示3个居群基于分子标记数据的遗传距离矩阵和基于果实及种子形态特征的欧氏距离矩阵之间的相关性不显著,表明果实与种子在居群间出现的表型分化除了受遗传因素影响外,还受到其它环境因子的强烈影响。     

青藏高原东缘西藏沙棘(Hippophae tibetana)果实性状变异

西藏沙棘是沙棘属(Hippophae L.)中分布于青藏高原和喜马拉雅高海拔地区的特有类群之一,其果实大小、形状、颜色以及条纹长等性状表现出了高度的多样性。对分布于青藏高原东缘西藏沙棘7个天然居群的果长、果宽、果形系数(果长/果宽)、果柄长、纹长等果实性状变异进行了统计分析,探讨了其果实变异水平及地理变异式样。结果表明,西藏沙棘5个果实性状在居群间和居群内均存在着极显著差异(p0.01)(除纹长在居群间存在着显著差异(p0.05)),居群间变异系数(CV)在11.19%~45.03%之间,果柄长的变异幅度最大(CV为45.03%),其次为纹长(CV为31.97%),变异幅度最小的是果宽(CV为11.19%);表型分化系数(Vst)在0.213 3~0.629 9之间,平均值为0.489 5,居群间和居群内变异约各占果实变异的一半。相关性分析显示,除果长与果柄、果宽与纹长外,其它各性状间均表现出极显著相关(p0.01),除果长与海拔、果宽与经纬度外,其它各性状与地理因子之间均达到极显著相关(p0.01),呈现出地理梯度变异,聚类分析表明7个居群按照地理距离远近聚类。     

Variability in reproductive traits in Jatropha curcas L. accessions during early developmental stages under warm subtropical conditions

Variability in floral, fruit, and seed characteristics, and oil content of 15 accession of Jatropha curcas during early development were assessed during two flowering periods in south Florida subtropical climate. The two flowering periods had leaf flushing in March. Field evaluation using 18 quantitative traits showed significant variation among accessions. The number of female flowers and female : male flower ratio ranged from 1 to 15 and 1 : 8.8 to 1 : 67.8, respectively. Fruit set by natural pollination was 89 and 66% during the first (1st) and second (2nd) flowering periods, respectively. A higher number of female‐type inflorescences were observed during summer. There were significant differences in seed traits, except for number of seeds per fruit. Accession TREC 31 had the highest individual seed dry weight and 100‐seed weight (0.83 g and 79.7 g, respectively). The oil content varied from 19.30% to 35.62%. Seed dry weight had positive correlation with seed fresh weight, seed length, seed thickness, seed width, and 100‐seed weight, but negative correlation with oil content. Based on the cluster analysis using 15 morphological traits, jatropha accessions were grouped into five main clusters and accessions from different geographic regions grouped together in a cluster. Principal component analyses (PCA) revealed morphological variation. The first three components explained 73.5% of the total variation and seed dry weight, 100‐seed weight, total flowers per inflorescence, male flowers per inflorescence and fruit set can be used to distinguish accessions. The PCA also indicated that flowering traits were more influenced by seed origin while seed traits were affected by flowering spans. Although evaluations were performed in plants during the juvenile phase, accessions TREC 31 and TREC 55 had superior averages for almost all characters evaluated. These results provide a preliminary assessment of the high variability in jatropha accessions evaluated and their potential for use in breeding and genetic improvement programs.     

Seed protein fraction electrophoresis in peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.) accessions and wild species

Total seed storage proteins were studied in 50 accessions of A. hypogaea (11 A. hypogaea ssp. hypogaea var hypogaea, 13 A. hypogaea ssp. hypogaea var hirsuta, 11 A. hypogaea ssp. fastigiata var fastigiata and 15 A. hypogaea ssp. fastigiata var. vulgaris accessions) in SDS PAGE. These accessions were also analysed for albumin and globulin seed protein fractions. Among the six seed protein markers presently used, it was found that globulin fraction showed maximum diversity (77.2%) in A. hypogaea accessions followed by albumin (52.3%), denatured total soluble protein fraction in embryo (33.3%) and cotyledon (28.5%). The cluster analysis based on combined data of cotyledons, embryos, albumins and globulins seed protein fractions demarcated the accessions of two subspecies hypogaea and fastigiata into two separate clusters supported by 51% bootstrap value, with few exceptions, suggesting the genotypes to be moderately diverse. Native and denatured total soluble seed storage proteins were also electrophoretically analysed in 27 wild Arachis species belonging to six sections of the genus. Cluster analysis using different methods were performed for different seed proteins data alone and also in combination. Section Caulorrhizae (C genome) and Triseminatae (T genome) formed one, distantly related group to A. hypogaea and other section Arachis species in the dendrogram based on denatured seed storage proteins data. The present analysis has maintained that the section Arachis species belong to primary and secondary genepools and, sections Procumbenetes and Erectoides belong to tertiary gene pools.     

Changes in fruit shape inproboscidea parviftora ssp.parviflora (martyniaceae) with domestication

Aboriginal cultivators selected a phylogenetically novel fruit shape during the domestication ofProboscidea parviflora ssp.parviflora var.Hohokamiana. The extremely long rostrum of the domesticated var.Hohokamiana is unique for the Martyniaceae. Rostrum length and width are significantly correlated in the wild var.parviflora, but in var.Hohokamiana aboriginal cultivators apparently selected for a weaker correlation between these variables. In both varieties there is an allometric relationship between rostrum and “capsule” lengths, with the former increasing proportionately faster than the latter. The above allometric relationship and the decrease in correlation between rostrum length and width are advantageous to aboriginal cultivators because the proportionately longer and thinner rostra are better for basketmaking. Comparisons of the data presented herein with data from other plants suggest that similar growth factors have been responsible for changes in fruit shape resulting from domestication.     

Characterization of genetic diversity of nuclear and mitochondrial genomes in Daucus varieties by RAPD and AFLP

The genetic diversity of nuclear genomes of five Daucus species and seven Daucus carota L. subspecies involving 26 accessions was characterized with random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). AFLP produced more than four times as many discrete bands per reaction compared with RAPD analysis, while both AFLP and RAPD basically led to similar conclusions. The dendrograms constructed with both RAPD and AFLP revealed that all accessions of D. carota were grouped into a major cluster delimited from other Daucus species, in good agreement with the classification by morphological char-acteristics. All accessions of cultivated carrots [(D. carota ssp. sativus (Hoffm.) Arcang.] were clustered in the same group while the variation within D. carota was relatively extensive. Genetic diversity of mitochondrial genomes was also documented with RAPD for the same accessions. The mitochondrial dendrogram differed from that of the nuclear genome, suggesting that nuclear and mitochondrial genomes of some accessions had separate evolutionary histories. Received: 20 September 1997 / Revision received: 19 January 1998 / Accepted: 28 March 1998     

New interspecific hybrids in Lathyrus (Leguminosae): Lathyrus annuus× L. hierosolymitanus

Experimental hybridization, cytogenetic observations, flower pigment analyses and scanning electron microscope observations of seed coat features were used to investigate the relationship between L. annuus L. and L. hierosolymitanus Boiss. Pod width, seed size, shape and surface ornamentation proved to be good diagnostic criteria for distinguishing the two taxa. Hybridization between various accessions of the two species revealed variation both between and within accessions. Enhanced anthocyanin levels were detected in the standard petals of the reciprocal hybrids made between two specific accessions of L. annuus and L. hierosolymitanus. The results of the investigations support the retention of L. annuus and L. hierosolymitanus as separate species.     

Comparative analysis of genetic structure and variability in wild and cultivated pomegranates as revealed by morphological variables and molecular markers

In this study, the genetic variability and relationships among wild and cultivated pomegranate genotypes from the north of Iran were investigated by morphological characters and RAPD molecular markers. Principal component analysis showed that the first three components explained 61.64 % of the total morphological variation for studied genotypes. Fruit neck diameter, anthocyanin index, TSS, aril juice, fruit flavor index, petiole length, fruit peel thickness and seed hardness were predominant in the first component and contributed most of the total variation. Fruit characteristics such as titratable acidity were negatively correlated (r = ?0.56) with TSS (r = ?0.56) and pH (r = ?0.86) and also, seed hardness showed negative correlation with aril length and aril diameter. Clustering from morphological data allocated individuals into two main clusters with high variation. Two hundred and twenty-nine fragments were scored of which 174 of them were polymorphic with 76.9 % polymorphism. Genetic similarity ranged from 0.15 to 0.78 with an average of 0.42, indicating high genetic variation among studied genotypes. High molecular and morphological variability indicated that this germplasm includes rich and valuable plant materials for pomegranate breeding.     

Classification of genetic diversity in Abelmoschus tuberculatus germplasm collection using morphometric data

Abelmoschus tuberculatus Pal & Singh is endemic to India and is the close relative of cultivated okra, A. esculentus. Data on 49 selected accessions of A. tuberculatus were used to study genetic diversity for a range of morphological characters through the use of multi-variate statistics. Epicalyx segments, shape, size and persistence; petal colour and petal blotch; stem and fruit pubescence; pigmentation of various plant parts; days to flowering; plant height; and the fruit yield/plant were the important components of variation and contributed significantly to the total variation in germplasm accessions under study. The accessions could be grouped into four distinct clusters. The grouping pattern, however, did not reflect relationship between geographic origin and genetic diversity. The potential use of the variability in okra improvement programmes is discussed.     

Pepper morphological traits related with resistance to Phytophthora capsici

Inheritance of 10 morphological and quantitative traits related to plant and fruit development and resistance to the pathogen Phytophthora capsici was studied in an intraspecific cross between a non-pungent, susceptible Capsicum annuum parent (cv. Americano) and a wild, pungent and resistant line (Serrano Criollo de Morelos-334). Data were obtained from the segregation of 166 F₂ plants and 50 F₃ plants in four years. Three of the traits analyzed (necrosis length, leaf width and leaf length) exhibited a transgressive segregation. A multiple linear regression analysis was applied in order to establish a relationship between necrosis length and some of the morphological traits measured such as length and width of leaf, length, diameter and mass of fruit, capsaicin content in fruits, and presence of hair on leaves and stems. The results identified a linear dependence between necrosis length (as an inverse measurement of resistance) and leaf width, fruit diameter and hair presence in the stem. Pungency was not related with resistance.     

Genetic diversity in traditional Ethiopian highland maize accessions assessed by AFLP markers and morphological traits

In the highland regions of Ethiopia the heterogeneity of the land, the climate, and soil favors the presence of a large number of landraces. We analyzed a representative sample of 62 traditional Ethiopian highland maize accessions, using amplified fragment length polymorphism (AFLP^®) markers and morphological traits with the aim to group the accessions based on the their genetic profiles and morphological traits, to study agroecological variation and to assess the level of correlation between phenotypic and genetic distances. Eight EcoRI/MseI primer combinations and 15 morphological traits were used. The accessions varied significantly for all of the measured morphological traits. Of a total of 650 AFLP markers that were scored, 89.5% were polymorphic. Pair-wise genetic distance estimates based on AFLP data revealed dissimilarity coefficients ranging from 0.32 to 0.69 (mean of 0.57). Cluster analysis of the AFLP data grouped most accessions collected from the Northern highlands into one major cluster. It, however, failed to separate the Western and Southern accessions into different clusters. Regardless of the large variation in environmental conditions between agroecologies, only 9% of the total genetic variation was found between agroecologies, whereas 91% was found within agroecologies in Ethiopia. This finding may be explained by long distance seed exchange, continuous seed introduction and gene flow between agroecologies. The relationship between morphological and AFLP-based distances was significant and positive. Based on this study, three groups of highland accessions, with distinctive genetic profiles and morphological traits were identified. This information will be useful for further collections and conservation of the unique diversity included in the highland maize landraces of Ethiopia.     

珍稀油料树种琴叶风吹楠果实和种子形态变异

以云南野生琴叶风吹楠(Horsfieldia pandurifolia)为材料,对39株树的果实和种子的形态进行变异分析。结果表明,果实和种子形态指标株间变异大,株内变异小。单株间果实表现出从卵圆形至长椭圆形的变异,果长4~7 cm,先端圆钝至锐尖,基部不同程度偏斜,与文献记录差异较大;单株间种子形态表现出从卵形、长卵形至细长形或近锥形的变异。单株平均种长变幅为25.05~35.66 mm,种宽为13.30~19.62 mm,长宽比为1.47~2.32,单因素方差分析表明此3个性状单株间差异极显著(p<0.01)。灰白色种皮具褐色斑块,种脐和胚位于种子基部,种子先端突尖或具短喙是稳定的性状,有重要的分类学价值。