Chromosome N-banding polymorphism in Aegilops geniculata Roth

The chromosome morphology and the variation in the N-band distribution along the chromosomes were studied in 13 accessions of the wild wheat Aegilops geniculata Roth (2n=28, genome formula UUM°M°). The karyotype consisted of four metacentric, five submetacentric, three subtelocentric, and two satellited pairs of chromosomes. The N-banding technique stained differentially all 14 chromosome pairs. The most prominent bands were observed near the centromeres and in the intercalary regions of both arms. Telomeric bands were found in only seven chromosomes. All chromosomes produced a specific banding pattern, permitting their identification. Polymorphism for presence or absence of particular bands was observed among the different accessions. The variable bands were located predominantly at the terminal and subterminal chromosome regions, and were also unevenly distributed over chromosomes. This polymorphism resulted in up to eight distinct banding pattern variants for each of the Ae. geniculata chromosomes. Each accession in this study showed a unique overall karyotype, sharing 0 to 8 chromosomes of identical banding pattern with the other accessions. A generalized idiogram of Aegilops geniculata is presented.     

Phenotypic responses of the wild wheat relative Aegilops geniculata Roth and wheat (Triticum durum Desf.) to experimentally imposed salt stress

In order to study the salt effect on the wild wheat and durum wheat, three accessions of Aegilops geniculata L. from Ain zana, Zaghouan, and Sbitla and one variety of wheat (Triticum durum L.) have been grown in the INRAT green house and treated with different salt concentrations. The morphological, phenological and yield characters have been measured for each plant and analyzed using SAS software. This study has shown a high degree of variation of these characters mainly related to geographical origin. It was observed also that Sbitla accession was less affected by the imposed salt stress than all the others while Ain zana was the most affected one.     

Conservation of the genetic variation of Triticum tauschii (Coss.) Schmalh. (Aegilops squarrosa auct. non L.) in synthetic hexaploid wheats (T. turgidum L. s.lat. x T. tauschii; 2n=6x=42, AABBDD) and its potential utilization for wheat improvement

Triticum tauschii (Coss.) Schmalh. (Aegilops squarrosa auct. non L., 2n=2x=14, DD genome) with its diverse range of accessions and distribution provides a unique opportunity for exploiting novel genetic variability for wheat (T. aestivum L.) improvement associated with biotic/abiotic stress factors. From our working collection of 490 T. tauschii accessions we have so far produced 430 different synthetic hexaploids (2n=6x=42, AABBDD) resulting from the chromosome doubling of Triticum turgidum L. s. lat. x T. tauschii F₁ hybrids (each synthetic involving a different T. tauschii accession). We present here our results on hybrid production, plantlet regeneration, cytology, colchicine induced doubling of the 2n=3x=21 chromosome F1 hybrids, seed increase of the doubled progeny and screening for a biotic stress; Cochliobolus sativus Ito and Kuribay (syn. Helminthosporium sativum Pamm. King and Bakke); of 250 of these synthetic hexaploid (2n=6x=42) amphiploids. Application of the direct crossing methodology involving susceptible T. aestivum cultivars with resistant T. tauschii accessions is also alluded to.     

Genetic diversity of Central Asian and north Caucasian Aegilops species as revealed by RAPD markers

RAPD analysis of 112 accessions of Aegilops tauschii Coss. (genome DD), Ae. cylindrica Host (CCDD), Ae. crassa Boiss. (DDMM), Ae. biuncialis Vis. (UUMM) and Ae. triuncialis L. (UUCC) collected in the Central Asia and north Caucasia was conducted. Aegilops accessions were divided into two major groups, corresponding to the D genome species and the U genome species. These groups were also separated into sub-groups according to species, except for the Ae. tauschii-cylindrica complex of accessions from Central Asia. Aegilops tauschii from north Caucasia was divided into two varietal groups, tauschii and meyeri. The Central Asian accessions of Aegilops species were more diverse than the accessions from north Caucasia. Aegilops tauschii and Ae. cylindrica accessions from north Caucasia were genetically uniform. Associations between altitudal variation of Aegilops species and variability of RAPD markers were not found.     

Interrelationships among Coconut (Cocos Nucifera L.) Accessions Using RAPD Technique

The genetic relationships among 33 coconut germplasm accessions were analyzed using RAPD markers. The germplasm accessions were collected from various coconut growing regions viz. South Asia (SA), South East Asia (SEA), South Pacific (SP), Atantic and America, and Africa. Forty-five random primers produced a total of 399 polymorphic markers. The Polymorphism Information Content (PIC) ranged from 0.031 to 0.392 and the Marker Index (MI) ranged from 6.28 to 0.031 among the primers. Based on the MI a set of 5, 10 and 15 informative and reproducible primers were identified. The mantel matrix correlation was calculated to compare the similarity matrices of a set of reproducible informative primers and global primers. There was significant correlation among the similarity matrices (r ≥ 0.50). The similarity matrix based on 399 polymorphic markers was used to construct the dendrogram to show the genetic relationship among the accessions. Similarity values ranged between 0.573 and 0.846. There was less genetic similarity (based on Jaccard's coefficient) among South Pacific and South East Asian accessions. The clustering pattern obtained in the present study was in agreement with the earlier reports based on RFLP, SSRs and AFLPs.     

Analysis of genetic diversity in Turkish sesame (Sesamum indicum L.) populations using RAPD markers⋆

The genetic diversity of 38 cultivated populations of Sesamum indicum L. from four different regions of Turkey was estimated at the DNA level with the random amplified polymorphic DNA (RAPD) technique. Sixty-one bands were obtained for all populations 78% of which were polymorphic. Analysis of molecular variance (AMOVA) was used to investigate the genetic diversity of the populations which yielded highly significant differences among populations within regions (91.9% of the total genetic diversity). According to AMOVA and Shannon's index that were performed separately for each region, the highest value of genetic variation was observed among Northwest region populations (CV = 7.7; H₀ = 0.304) and lowest in the Southeast regions' populations (CV = 2.6; H₀ = 0.068). Nei and Li's similarity index was calculated and phylogenetic tree was established using the neighbor-joining algorithm. This phenetic analysis grouped 35 of 38 accessions in six groups leaving three highly diverse accessions outside. Wagner phylogenetic method was used to assess the phylogenetic relationships among the populations. In the majority-rule consensus tree, only 7 of the 32 forks showed above 60% occurrence. Using Principal Coordinate Analysis (PCO) of the RAPD data set, the groups were clearly separated along the first three axis. These results indicate that RAPD technique is useful for sesame systematics, and should be valuable for the maintenance of germplasm banks and the efficient choice of parents in breeding programs.     

Isozyme variation in species of the section Comopyrum of Aegilops

Genetic variation at 21 enzyme loci was studied in Aegilops comosa and Ae. uniaristata, the two species belonging to section Comopyrum of Aegilops. In Ae. comosa, the mean number of alleles per locus was 2.00 and the proportion of polymorphic loci was 0.667; in Ae. uniaristata they were 1.19 and 0.143, respectively. The two species were genetically distant from each other (I=0.561) supporting the previously assigned different genome symbols, M and N.     

Genetic diversity analysis in Boerhavia diffusa L. of different geographic locations in India using RAPD markers

Boerhavia diffusa is extensively used in herbal medicines as well as in the Ayurvedic system, because it contains a set of clinically important compounds. In the present study, the genetic variability in Boerhavia diffusa between accessions of different geographical origin within the Indian territory is assessed through random amplified polymorphic DNA (RAPD) markers. Twenty-eight accessions of Boerhavia were screened with eighteen primers of which nine were found to be the most informative. The degree of polymorphism was found to be high in accessions collected from different places of Uttar Pradesh (Set II) in comparison to other states of India (Set I). A relatively lower level of polymorphism was recorded in accessions collected from diverse locations around Lucknow (Set III). Accessions from neighbouring geographical regions exhibited more similarity than those from distant regions (as revealed by the set I analysis). Certain diagnostic markers may be correlated with morphological character(s) such as plant type. BDL appeared most distinct and divergent from the rest of the accessions and the BDJ plant in set II also showed least similarity estimate. Fragments of 5.62 Kb and 4.47 Kb with primer GN59 was found to be unique for BDP and BD2 having ovate leaf character, whereas ovoid leaf genotype exhibited 0.79 Kb (GN34 primer) fragment. Similarly a unique band type (0.35 Kb) with primer GN83 was present in BDL and BD1 that share light pink flower. Jaccard's, and Nei and Li similarity coefficient values amongst the accessions were in the range of 0.22 to 0.89 and 0.33 to 0.93, respectively. Association of RAPD markers with the leaf characteristics, flower colour as well as with geographical locations has been made. This shows that RAPD markers are also useful for the study of genetic structure of Boerhavia populations.     

Genetic diversity in cowpea [Vigna unguiculata (L.) Walp.] as revealed by RAPD markers

The present study, using RAPD analysis, was undertaken to characterize genetic variation in domesticated cowpea and its wild progenitor, as well as their relationships. The materials used consisted of 26 domesticated accessions, including accessions from each of the five cultivar-group, and 30 wild/weedy accessions, including accessions from West, East and southern Africa. A total of 28 primers generated 202 RAPD bands. One hundred and eight bands were polymorphic among the domesticated compared to 181 among wild/weedy cowpea accessions. Wild accessions were more diverse in East Africa, which is the likely area of origin of V. unguiculata var. spontanea. Var. spontanea is supposed to have spread westward and southward, with a loss of variability, loss counterbalanceed in southern Africa by introgressions with local perennial subspecies. Although the variabilty of domesticated cowpea was the highest ever recorded, cultivar-groups were poorly resolved, and several results obtained with isozyme data were not confirmed here. However primitive cultivars were more diverse than evolved cultivars, which still suggests two consecutive bottlenecks within domesticated cowpea evolution. As isozymes and AFLP markers, although with a larger number of markers, RAPD data confirmed the single domestication hypothesis, the gap between wild and domesticated cowpea, and the widespread introgression phenomena between wild and domesticated cowpea.     

Powdery mildew resistance in Aegilops tauschii coss. and synthetic hexaploid wheats

Summary A collection of 400 Ae. tauschii (syn. Ae. squarrosa) Coss. accessions were screened for powdery mildew resistance based on the response patterns of 13 wheat cultivars/lines possessing major resistance genes to nine differential mildew isolates. 106 accessions showed complete resistance to all isolates, and 174 accessions revealed isolate-specific resistance, among which were 40 accessions exhibiting an identical response pattern as wheat cultivar Ulka/^*8Cc which is known to possess resistance gene Pm2. Expression of both complete and isolate-specific resistance from Ae. tauschii was observed in some synthetic hexaploid wheats derived from four mildew susceptible T. durum Desf. parents, each crossed with five to 38 resistant diploid Ae. tauschii accessions. Synthetic amphiploids involving different combinations of T. durum and Ae. tauschii generally showed a decrease in resistance compared with that expressed by the Ae. tauschii parental lines.     

Distribution and diversity of Aegilops tauschii in Iran

The wide morphological variation of Aegilops tauschii has led to the distinction of different subspecies; a typical ssp. tauschii and a second ssp. strangulata. However some researchers pointed out the existance of the intermediate form among morphologically distinguished subspecies. Distribution, diversity and the relationship between different subspecies and the intermediate form were evaluated in the Iranian Ae. tauschii collection. This collection was classified to 15 different populations according to morphological similarities and the collecting origin of accessions. The highest variation was found in tauschii population of Golestan followed by tauschii populations of Gilan and Ardebill, whereas the lowest variation was observed in tauschii populations of central Iran. Two discriminant functions suggested that the length of rachis node and spikelet glume, particularly, the length/width ratios of these traits had the highest impact on identification of different forms. Mahalanobis distances (D ² ) between the two subspecies along with intermediate form on the multidimensional scaling plot showed that the intermediate form is more similar to ssp. tauschii than ssp. strangulata. Although, the diversity within the ssp. strangulata was not very high, it widely affected the diversity of Iranian accessions of Ae. tauschii through continues crossing with the more diversed subspecies, tauschii, during thousands of years. This fact had lead to expansion of its distribution from its origin to Northern Khorasan, Northern Semnan and Eastern Ardebill by producing the intermediate form.     

Genetic variation of Ethiopian and Eritrean sorghum (Sorghum bicolor (L.) Moench) germplasm assessed by random amplified polymorphic DNA (RAPD)

The extent and patterns of distribution of genetic variation among 80 sorghum (Sorghum bicolor (L.) Moench) germplasm accessions from Ethiopia and Eritrea were investigated using RAPD with 20 oligonucleotide primers. The primers generated a total of 147 polymorphic bands across the 80 accessions with a mean of 7.35 bands per primer. Estimation of the extent of variation by the Shannon-Weaver diversity index revealed an intermediate level of overall variation (H = 53), although the levels varied among regions of origin of the accessions. Partitioning of the total variation revealed considerable variation (77%) within the region of origin of the accessions and the remainder (23%) among regions of origin. Similarly, a large portion (94%) of the total variation was found within the adaptation zones compared to among the adaptation zones (6%). The results suggest a weak differentiation of the sorghum material both on regional and agro-ecological bases, which could be ascribed to the high rate of outcrossing in cultivated sorghum and its free natural hybridization with its wild and weedy relatives, as well as to seed movement by humans. The average genetic dissimilarity was found to be 36% among the 80 accessions and 13% among the 15 regions of origin. Cluster analysis failed to group accessions of the same region or the same adaptation zone, which further confirmed the weak differentiation of the material studied. The clustering pattern of the regions of origin was broadly concordant with previous clustering patterns obtained using morphological characters, in which regions with broad agro-climatic conditions were grouped together.     

Resistance to wheat leaf rust in Aegilops tauschii Coss. and inheritance of resistance in hexaploid wheat

A collection of 164 Aegilops tauschii accessions, obtained from Gatersleben, Germany, was screened for reaction to leaf rust under controlled greenhouse conditions. We have also evaluated a selection of synthetic hexaploid wheats, produced by hybridizing Ae. tauschii with tetraploid durum wheats, as well as the first and second generation of hybrids between some of these resistant synthetic hexaploid wheats and susceptible Triticum aestivum cultivars. Eighteen (11%) accessions of Ae. tauschii were resistant to leaf rust among which 1 was immune, 13 were highly resistant and 4 were moderately resistant. Six of the synthetic hexaploid wheats expressed a high level of leaf rust resistance while four exhibited either a reduced or complete susceptibility compared to their corresponding diploid parent. This suppression of resistance at the hexaploid level suggests the presence of suppressor genes in the A and/or B genomes of the T. turgidum parent. Inheritance of leaf rust resistance from the intercrosses with susceptible bread wheats revealed that resistance was dominant over susceptibility. Leaf rust resistance from the three synthetics (syn 101, syn 701 and syn 901) was effectively transmitted as a single dominant gene and one synthetic (syn 301) possessed two different dominant genes for resistance.     

Biodiversity of Diploid D-Genome Aegilops Tauschii Coss. in Iran Measured Using Microsatellites

Microsatellite markers were used to analyse the biodiversity of 57 accessions of different subspecies and varieties of wild Aegilops tauschii (2n = 2x = 14; D genome) collected across the major areas where it grows in Iran. Levels of diversity were high, with numbers of alleles averaging 7.3 (ranging up to 12) and polymorphism information contents averaging 0.6591. One accession was notably more similar to two of the D genome in hexaploid wheats (Triticum aestivum) used as outgroups. Within the Ae. tauschii accessions, no markers were characteristic for taxa or geographical origin, suggesting high gene flow between the subspecies and varieties, although some groupings, which could be related to geographical origin, were evident. This survey demonstrates the high diversity present in wild goatgrass in Iran, and indicates that there is value in sampling for useful genes for wheat breeding.     

Aegilops tauschii: Genetic Variation in Iran

All the 79 Aegilops tauschii Coss. accessions of Iranian origin from Prof. Kihara’s collection were analyzed electrophoretically. Of 23 enzyme-encoding loci studied, 11 were polymorphic. In Iran Ae. tauschii is presented by ssp. tauschii and ssp. strangulata which distinctly differ genetically, morphologically and ecologically. Variation patterns of low polymorphic locus Aco2 and highly polymorphic Ep are similar in both subspecies. In contrast, variation of Acph1, Ak, Est2, Est5, Got1, Got2, Got3 and Lap is a set of diverse patterns which markedly differ between subspecies and natural regions also, implying that natural selection is involved.     

用RAPD分析辣椒细胞质雄性不育基因

根据近等基因系分析原理 ,以辣椒细胞质雄性不育系 93 A及其保持系 93 B为材料 ,对辣椒细胞质雄性不育基因及其保持基因开展了RAPD标记研究 ,结果表明 :OPK 1 7550 、OPK 1 7150 0 标记可能与细胞质雄性不育基因相连锁 ,OPH 1 1 2 0 0 0 标记可能与保持系中的保持基因相连锁。对侯选标记OPK 1 7550 进行了克隆和部分序列分析 ,为利用混合群体分离法 (BSA)对分离群体中的不同单株进行SCAR分析的标记验证工作奠定了基础。     

Identification and overcoming barriers between Brassica rapa L. em. Metzg. and B. nigra (L.) Koch crosses for the resynthesis of B. juncea (L.) Czern.

Brassica juncea (2n = 36, AABB) is an amphidiploid derived from its diploid progenitor species B. rapa (2n = 20, AA) and B. nigra (2n = 16, BB). Resynthesis of B. juncea by exploiting the wider gene pool of the present day diploids offers a powerful tool for obtaining novel genetic variation. Hybridization between the two species is difficult because of the presence of crossability barriers. Barriers to hybridization were identified using Aniline Blue Fluorescence (ABF) method. Crosses were classified as having pre- or post-fertilization barriers based on whether less than or more than one per cent ovules showed pollen tube entry. Of the 23 crosses studied, seven crosses all with B. rapa as the female parent showed pre-fertilization barriers. Only 0 to 0.3 per cent ovules were fertilized in these crosses. The remaining 16 crosses, four with B. rapa as the female parent and 12 with B. nigra as the female parent, showed post-fertilization barriers. In crosses with pre-fertilization barriers use of bud pollination and stump pollination ensured or increased fertilization. These two simple techniques were used in combination with ovary-ovule culture to ensure recovery of hybrids. In crosses with post-fertilization barriers, ovary-ovule culture alone helped to obtain hybrids. Hybrids were obtained at higher frequency with the use of direct ovule culture compared to ovary-ovule culture. The F₁ plants obtained in vitro were multiplied on MS medium + 2 mg/l Kn + 0.2 mg/l IAA. These F₁ plants were confirmed as true hybrids through morphology, leaf isozymes and cytology. In all 17 hybrids were obtained of which three were amphidiploids.     

Evaluation for resistance to Pyrenophora tritici-repentis in Aegilops tauschii Coss. and synthetic hexaploid wheat amphiploids

Summary A total of 59 diploid Aegilops tauschii Coss. (syn. Aegilops sguarrosa auct. non L.) and 39 synthetic hexaploid wheat accessions were evaluated for reaction to Pyrenophora tritici-repentis (Died.) Drechs. in a controlled environment, and classified using a disease rating system based on lesion type. 27 Ae. tauschii and 20 synthetic wheats were found to be resistant to tan spot disease. The overall mean disease ratings of Ae. tauschii and the synthetic wheat lines scored on a scale of 1 (resistant) to 5 (susceptible) were 1.80 and 2.38, respectively. Synthetic wheats generally showed a decrease in resistance, although several lines of synthetic wheat expressed a higher resistance than the diploid parents. Five synthetic wheat lines exhibited higher resistance than the standard resistant common wheat cultivar Red Chief.     

Analysis of genetic diversity in Indian taro [Colocasia esculenta (L.) Schott] using random amplified polymorphic DNA (RAPD) markers

Taro [Colocasia esculenta (L.) Schott] germplasm accessions collected from different parts of India were subjected to RAPD (Random Amplified Polymorphic DNA) analysis to assess the genetic diversity prevalent and also to test the genetic basis of morphotypic classification. Thirteen random decamer primers out of the 22 tested were used to analyse 32 taro accessions belonging to 28 morphotypes. Three out of these thirteen primers analysed showed 100 per cent polymorphism. Per cent polymorphism varied from 60 to 100 among the polymorphic primers. High genetic diversity was revealed as the similarity coefficient values ranged from 0.50 to 0.98. No two accessions analysed in the present study showed a similarity coefficient value of one thereby indicating their distinctness and presence of high genetic diversity in Indian taro germplasm. Dendrogram obtained from UPGMA analysis grouped 32 accessions in four clusters and three accessions were placed as outliers. Clustering pattern did not show any strict relationship with geographical distribution, morphotype classification and genotypic diversity. Further, accessions classified, as belonging to the same morphotypic group did not always cluster together. Presence of a very close genepool of the wild, weedy and cultivated forms with extreme levels of phenotypic and genotypic variation is suggested as the reason for high genetic diversity reported. Usefulness of DNA markers such as RAPD in characterising and assessing the genetic diversity in Indian taro germplasm is hereby demonstrated.     

RAPD and seed coat morphology variation in annual and perennial species of the genus Cicer L.

Molecular and morphological variation of six perennial and five annual species including domesticated chickpea, C. arietinum, were inferred on the basis of RAPD and S.E.M. seed coat features using three outgroup taxa (Lens ervoides, Lathyrus japonica and Pisum sativum). Of the 66 decamer arbitrary primers tested, eight primers revealed 87 informative fragments. Neighbor-joining cluster analysis using Jaccard's coefficient of similarity on the basis of polymorphic fragments indicated a narrow variation in C. arietinum and recognized two major clusters in the genus Cicer. The first one included four species of sect. Monocicer: C. echinospermum, C. arietinum, C. reticulatum and Iranian material of C. bijugum. The second cluster contained annual and perennial species belonging to sections Chamaecicer, Polycicer and Acanthocicer. The character state of morphological and ecological traits on the RAPD phenogram indicated no monophyletic incision. Our results suggested that the high genetic difference between annual and perennial species might be regarded as a rapid speciation of section Monocicer. Reconsideration of traditional classification of sections Polycicer and Acanthocicer is necessary. The Desi and Kabuli types of C. arietinum could not be separately grouped at the DNA level, and the low level of genetic variation of C. arietinum may result from a bottleneck during the domestication process.