Genetic relationships among perennial and annual<Emphasis Type="Italic"> Cicer</Emphasis> species growing in Turkey assessed by AFLP fingerprinting

AFLP markers were used to assess genetic relationships among Cicer species with distribution in Turkey. Genetic distances were computed among 47 Cicer accessions representing four perennial and six annual species including chickpea, using 306 positions on AFLP gels. AFLP-based grouping of species revealed two clusters, one of which includes three perennial species, Cicer montbretii, Cicer isauricum and Cicer anatolicum, while the other cluster consists of two subclusters, one including one perennial, Cicer incisum, along with three annuals from the second crossability group (Cicer pinnatifidum, Cicer judaicum and Cicer bijugum) and the other one comprising three annuals from the first crossability group (Cicer echinospermum, Cicer reticulatum and Cicer arietinum). Consistent with previous relationship studies in the same accession set using allozyme and RAPD markers, in AFLP-based relationships, C. incisum was the closest perennial to nearly all annuals, and C. reticulatum was the closest wild species to C. arietinum. Cluster analysis revealed the grouping of all accessions into their distinct species-clusters except for C. reticulatum accessions, ILWC247, ILWC242 and TR54961; the former was found to be closer to the C. arietinum accessions while the latter two clustered with the C. echinospermum group. Small genetic distance values were detected among C. reticulatum accessions (0.282) and between C. reticulatum and C. arietinum (0.301) indicating a close genetic similarity between these two species. Overall, the AFLP-based genetic relationships among accessions and species were congruous with our previous study of genetic relationships using allozymes. The computed level of AFLP variation and its distribution into within and between Cicer species paralleled the previous report based on RAPD analyses. AFLP analysis also confirmed the presence of the closest wild relatives and previous projections of the origin of chickpea in southern Turkey. Results presented in this report indicate that AFLP analysis is an efficient and reliable marker technology in determination of genetic variation and relationships in the genus Cicer. Obviously, the use of AFLP fingerprinting in constructing a detailed genetic map of chickpea and cloning, and characterizing economically important traits would be promising as well.Communicated by P. Langridge     

iPBS-Retrotransposons-based genetic diversity and relationship among wild annual Cicer species

Lack of requisite genetic variation in cultivated species has necessitated systematic collection, documentation and evaluation of wild Cicer species for use in chickpea variety improvement programs. Cicer arietinum has very narrow genetic variation, and the use of a wild relative in chickpea breeding could provide a good opportunity for increasing the available genetic variation of cultivated chickpea. Genetic diversity and the relationship of 71 accessions, from the core area of chickpea origin and domestication (Southeastern Turkey), belonging to five wild annual species and one cultivated species (Cicer arietinum) were analysed using iPBS-retrotransposon and ISSR markers. A total of 136 scorable bands were detected using 10 ISSR primers among 71 accessions belonging to 6 species, out of which 135 were polymorphic (99.3 %), with an average of 13.5 polymorphic fragments per primer, whereas iPBS detected 130 bands with 100 % polymorphism with an average of 13.0 bands per primer. C. echinospermum and C. pinnatifidum were the most diverse among species, whereas C. arietinum exhibited lower polymorphism. The average polymorphism information contents (PIC) value for both marker systems was 0.91. The clustering of the accessions and species within groups was almost similar, when iPBS and ISSR NeighborNet (NNet) planar graphs were compared. Further detailed studies are indispensable in order to collect Cicer germplasm, especially C. reticulatum, from southeastern Turkey particularly, from Karacada? Mountain for preservation, management of this species, and to study their genetic diversity at molecular level. This study also demonstrates the utility and role of iPBS-retrotransposons, a dominant and ubiquitous part of eukaryotic genomes, for diversity studies in wild chickpea and in cultivated chickpea.     

Analysis of the subunit structure of protochlorophyllide holochrome by sodium dodecyl sulfate-polyacrylamide gel electrophoresis

The subunit structures of protochlorophyllide holochrome (PCH) and chlorophyllide holochrome (CH) were studied by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. PCH from leaves of dark-grown (Phaseolus vulgaris var. red kidney) is a polymeric pigment-protein complex of approximately 600,000 daltons. It is composed of 12 to 14 polypeptides of 45,000 daltons, when examined prior to and immediately following photoconversion. The protochlorophyllide or chlorophyllide pigment molecules are associated with these polypeptides. Subsequent to photoconversion, the absorption maximum of newly formed chlorophyllide shifts from 678 nm to 674 nm upon standing in darkness. Following the 678 to 674 spectral shift, the chlorophyllide is associated with a polypeptide with a molecular weight of 16,000 daltons. In addition, sucrose gradient centrifugation of PCH and CH under nondenaturing conditions indicates that during the course of the dark spectroscopic shift, the 600,000 dalton CH undergoes dissociation into a small chlorophyllide protein. The dissociation of CH, the change in the molecular weight of the chlorophyllide polypeptide from 45,000 to 16,000 daltons, as well as the dark spectroscopic shift are temperature-dependent and blocked below 0 C. It was also found that each holochrome molecule of 600,000 daltons contains at least four protochlorophyllide pigment molecules.     

Outer membrane proteins of Escherichia coli. II. Heterogeneity of major outer membrane polypeptides

When E. coli outer membrane protein is dissolved in sodium dodecyl sulfate (SDS) solution and boiled briefly, a single major peak (peak B) with a molecular weight of 42,000 daltons is observed on SDS-containing polyacrylamide gels. If the protein is dissolved in SDS solution at 37 °C and applied to gels without further treatment, peak B disappears and two other major peaks appear: Peak A, which is composed of aggregates and migrates more slowly than peak B, and peak C which is composed of monomeric protein not fully reacted with SDS and which migrates faster than peak B. When cyanogen bromide peptides of protein from peak A and peak C were compared, it was evident that peak A and peak C contained entirely different polypeptides. This was further confirmed by differential labeling studies with methionine and leucine. The cyanogen bromide peptide profiles of protein from peak A suggested that this peak was composed of two polypeptides, and this was confirmed by electrophoresis in an alkaline gel system which resolves peak B into three subcomponents. Two of these were derived from peak A and the third was derived from peak C. These results indicate that the outer membrane of E. coli contains at least three nonidentical major polypeptides, each of which has a nearly identical molecular weight of about 42,000 daltons. These polypeptides are present in identical proportions in the soluble and insoluble fractions obtained when the outer membrane is treated with Triton X-100 plus EDTA.     

A note on the genusBrassavola (Orchidaceae)

This paper contains a brief survey of the four subgeneric sections which comprise the genusBrassavola R. Br. of theOrchidaceae, with special reference to the group of species (§Grandiflorae Rolfe) whichSchlechter proposed separating fromBrassavola as the genusRhyncholaelia. The author maintains that these species should be allowed to remain inBrassavola as a distinct section.     

Isozyme polymorphism and phylogenetic interpretations in the genus Cicer L.

Summary Allozyme variation among 50 accessions representing the cultivated chickpea (Cicer arietinum L.) and eight wild annual Cicer species was scored and used to assess genetic diversity and phylogeny. Sixteen enzyme systems revealed 22 putative and scorable loci of which 21 showed polymorphism. Variation was prevalent between species (D_st = 0.510) but not within species (H_s = 0.050). No variation for isozyme loci was detected in the cultivated chickpea accessions. Cicer reticulatum had the highest proportion of polymorphic loci (0.59) while the loci Adh-2 and Lap were the most polymorphic over all the species accessions. The phylogeny of annual Cicer species, as determined by allozyme data, generally corroborated those based on other characters in previous studies. Cicer arietinum, C. reticulatum and C. echinospermum formed one cluster, while C. pinnatifidum, C. bijugum and C. judaicum formed another cluster. Cicer chorassanicum was grouped with C. yamashitae, whereas C. cuneatum formed an independent group and showed the largest genetic distance from C. arietinum.     

Photolabeling of glucose-sensitive cytochalasin B binding proteins in erythrocyte, fibroblast and adipocyte membranes

(³H)Cytochalasin B has been photoincorporated into membrane fractions of the human erythrocyte, Rous sarcoma virus-transformed chicken embryo fibroblast and rat adipocyte. Identification of D-glucose sensitive cytochalasin B binding sites was achieved by photolyzing membranes with radioligand in the presence of 0.5–0.7M D- or L-glucose. In the erythrocyte the major labeled bands on SDS-polyacrylamide gels were at 55,000 and 46,000 daltons. In the virus-transformed fibroblasts a major labeled band was at 55,000 daltons, and in adipocyte microsomal membranes, peaks at 50,000 and 45,000 daltons were observed. Binding characteristics of these polypeptides suggest that they are the putative glucose transport proteins in these three cell types.     

New cultivated species of the genusCryptocoryne (Araceae)

Three new species ofCryptocoryne are described, based on material received from unspecified localities in tropical Asia.Crytocoryne hejnýi Rataj andC. zukalii Rataj differ fromC. cor data Griff. by the non-infundibuliform gorge of the upper tube and by the colour of the limb of spathe.C. evae Rataj is one of the largest species of the genus and is related toC. siemensis Gagn, andC. zonata De Wit. It differs fromC. siemensis andC. blasii by the limb of spathe which is twice as broad and long, and by the coloured inner part of the kettle, fromC. zonata by the shape of the leaf-blades.     

Variation patterns within the genusCarex L. sect.Digitatae (Fries) Christ in Bulgaria

Seventeen natural Bulgarian populations of the species of the genusCarex sect.Digitatae were studied morphologically and karyologically. Stepwise discriminant analysis was used as a multivariate test for the separation of groups of populations from different taxa and to find a subset of characters contributing most for this separation. The following taxa were found in Bulgaria:C. humilis Leysser var.humilis; C. humilis var.longifolia Stoeva etPopova, var. n.;C. digitata L.;C. ornithopoda Willd. subsp.bulgarica (Vel.) Stoeva etPopova, comb. n. Aneuploid series of some chromosome numbers were found inC. digitata — 2n=48, 50, 52, 54, 56 and inC. ornithopoda subsp.bulgarica?2n=52, 53, 54, 56.C. humilis has 2n=36 and a much more heterogeneous karyotype than that of the above taxa. The values of the Euclidean distance between the populations vary within similar limits in both speciesC. humilis andC. digitata; this is not in accordance with the karyological results. The within and between population morphological variations were compared in each species. The first dominates in overall variation, which is probably connected with the very large populations and their mosaic space structure.     

LignicolousHyphomycetes from Czechoslovakia 6.Spadicoides andDiplococcium

Seventeen species belonging toSpadicoides Hughes andDiplococcium Grove collected in Central Europe mostly in Czechoslovakia are described and illustrated. Two new species are described inSpadicoides—S. carpatica Hol.-Jech.,S. penatium Hol.-Jech. and three new species inDiplococcium—D. bicolor Hol.-Jech.,D. insolitum Hol.-Jech. andD. parcum,Hol.-Jech.     

New fig insects [Hym.: Chalcidoidea] from India: Three torymids parasitic on agaonids

Two new species of fig wasps (Sycoscapter punctatus Abdurahiman & Joseph andSycobiella amplissima Abdurahiman & Joseph) parasitic onManiella delhiensis Abdurahiman & Joseph fromFicus amplissima Smith, and the female of a known species (Philotrypesis anguliceps Westwood) parasitic onBlastophaga quadraticeps Mayr fromFicus religiosa L. are described.     

Relative Embryo Growth Rates in the Annual Cicer L. Species

Early embryo growth rates were studied in the nine annual speciesof Cicer L., namely, C. arietinum L., C. bijugum Rech., C. chorassanicum(Bge.) M. Pop., C. cuneatum Rich., C. echinospermum Dav., C.judaicum Boiss, C. pinnatifidum J. and S., C. reticulatum Lad.and C. yamashitae Kit. The number of embryo cells increasedexponentially with time and was log linear in all the species.Species differed in their mean cell doubling time (MCDT). Cicerechinospermum and C. yamashitae had, respectively, the longestand the shortest MCDT which ranged from 9.67 to 16.15 h forthe nine species. Failure of successful interspecific hybridizationbetween C. arietinum and the wild annual species was only partlyexplained by differences in MCDT of the parental species. Relativegenetic closeness still plays the major role in determiningsuccess of interspecific hybridization in Cicer. Chickpea, Cicer, embryo, interspecific hybridization, suspensor     

On the electrotransfer of polypeptides from gels to nitrocellulose membranes

The conditions which affect the elution of polypeptides from polyacrylamide gels by electrophoresis and polypeptide-nitrocellulose interactions have been studied. The rate of elution of polypeptides from a 15% sodium dodecyl sulfate-polyacrylamide gel is dependent on the molecular weight of the individual polypeptides, which is in agreement with the results of W. N. Burnette (Anal. Biochem. 112, 195 (1981)). We also observed that current density affects the rate of elution. Polypeptides smaller than 20,000 daltons pass through pores of 0.45 microns, but not through the pores of 0.1-microns nitrocellulose membranes during electrophoresis. The nonionic detergent NP-40 inhibits the binding of polypeptides to nitrocellulose and removes prebound polypeptides from the membranes. Amido black and Coomassie blue staining and destaining processes do not remove the bound polypeptides from the membranes, but may affect the antigenicity of polypeptides. Polypeptides immobilized on nitrocellulose can be stored at -70 degrees C for future use.     

Genetic relationships in the genus Cicer L. as revealed by polyacrylamide gel electrophoresis of seed storage proteins

Summary Total seed storage protein of the cultivated chickpea, C. arietinum L., and eight other wild annual Cicer species (all 2n = 16) was separated and compared by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The seed-protein profile was a conservative and species-specific trait. Relative interspecific similarities of protein patterns were estimated using Jaccard's similarity index, and a cluster analysis was performed. The resultant dendrogram generally agreed with the limited data already available on interspecific relationships in Cicer based on morphological characteristics, crossability, genome pairing in hybrids, karyotypes and isozyme analysis. The difference between the profiles of C. judaicum and C. pinnatifidum supported the idea that they are indeed two separate species. The closest relative of C. arietinum was C. reticulatum, followed by C. echinospermum and other species, while C. cuneatum was the farthest relative. In general, C. cuneatum was also genetically the farthest removed from any other species. The suggestion that C. reticulatum is the wild progenitor of the cultivated chickpea was therefore further supported.     

Genetic relationships among the annual species of Cicer L.

Summary Genetic relationships between 7 annual species of the genus Cicer, including the cultivated chickpea, have been studied. These species were assigned to 3 crossability groups. In each group interspecific hybrids could be obtained but their fertility differed considerably in the various cross combinations. Crosses between members of different groups yielded no viable seeds. The possibility of gene transfer from the wild species to the cultivated chickpea C. arietinum was also assessed. Only two species could be considered for this purpose, C. reticulatum, which is the wild progenitor of the cultivated species, and C. echinospermum, which is in the secondary gene pool of C. arietinum. A unique postzygotic reproductive barrier mechanism was found between the members of Group II, C. judaicum, C. pinnatifidum and C. bijugum. It is based on a disharmony in the growth rate of the stigma and the anthers at the time of anthesis of the F₁ interspecific hybrid so that selfpollination is avoided. It is proposed that this kind of mechanism has been involved only when an effective spatial isolation between the three species had been obtained.     

Duration of development and longevity inAphidius ervi andAphidius platensis [Hym.: Aphidiidae], two parasites ofMyzus persicae [Hom.: Aphididae]

Rate of development and longevity were studied inAphidius ervi Haliday andAphidius platensis Brèthes, two parasites of the green peach aphid,Myzus persicae Sulz. On paprika,A. ervi developed from egg to adult in 27.3 days (at 15°C) and 19.9 days (21°C),A. platensis in 19.9 days (15°C), 15.6 days (21°C) and 12.4 days (24°C). The period from oviposition to mummification was in both species roughly twice as long as the period from mummification to adult emergence. Males emerged slightly before females. Given water and honey,A. ervi lived 15.4 days (♂) and 13.1 days (♀) at 21°C. The effect of temperature on longevity was tested inA. platensis on this diet: at 15°C, 9.2 days (♂) and 7.9 days (♀); at 21°C, 12.0 days (♂) and 13.4 days (♀); and at 24°C, 7.4 days (♂) and 8.4 days (♀). When supplied only with water, both species lived for 1–3 days. When aphid-infested leaves were added, longevity increased by 3.5 days (A. platensis). The maximal longevity, obtained with water and honey, was somewhat reduced when leaves were added, probably due to mating and oviposition activities (A. ervi). Longevity was not significantly influenced by the different host plants during parasite development. Differences in longevity between the sexes were small and dependent on temperature and food.     

Ploidy variation in the group ofMyosotis palustris andM. laxa in the Czech Republic and Slovakia

In 74 natural populations of theMyosotis palustris andM. laxa groups from former Czechoslovakia three euploid and one aneuploid cytotypes were found that belong to the following species:M. nemorosa Besser, 2n=22,M. palustris (L.)Nath. em.Reichenb. inSturm, 2n=66,M. laxiflora Reichenb., 2n=66,M. radicans Opiz, 2n=66(64),M. brevisetacea (Schuster) Holub, 2n=66(64), andM. caespitosa C.F. Schultz, 2n=88. With the exception ofM. nemorosa, the chromosome numbers are published for the first time for the Czech Republic. The chromsome numbers, 2n=66, 2n=64 are found inM. brevisetacea, are new for the literature. The correlation between pollen size and ploidy level has been studied as well, and statistically significant differences were found between cytotypes belonging to different ploidy levels.     

Chromosome counts and chromosome morphology of some selected species

In the years 1976–1981 we studied chromosome counts and karyotypic formulae of the following 29 species of plants from 41 localities (of these 6 from Bohemia, 32 from Moravia, 3 from Slovakia):Batrachium baudotii (Godron) F. W. Schultz,Chenopodium rubrum L.,C. polyspermum L.,C. murale L.,C. ficifolium Sm.,C. opulifolium Schrader ex DC. inLam. et DC.,C. strictum Roth [subsp.strictum, subsp.glaucophyllum (Aellen)Aellen inJust etAellen, subsp.striatiforme Uotila],Arenaria grandiflora L.,Illecebrum verticillatum L.,Spergula morisonii Boreau inDuchartre,Spergularia marginata (DC. inLam. et DC.)Kittel S. marina (L.)Griseb.,S. rubra (L.) J. etC. Presl,Silene conica L.,Sisymbrium loeselii L.,S. volgense Bieb. exE. Fourn.,S. orientale L. [subsp. orientale, subsp.macroloma (A. Pomel)Dvo?ák],S. officinale (L.)Scop.,Descurainia sophia (L.)Webb exPrantl inEngler etPrantl,Nasturtium officinale R. Br. inAiton,Barbarea arcuata (Opiz inPresl J. et C.)Reichenb.,Lunaria annua L.,Soldanella montana Willd.,S. carpatica Vierh. inUrban etGraebner,Lotus tenuis Waldst. etKit. exWilld.,L. uliginosus Schkuhr,Trigonella monspeliaca L.,Geranium sibiricum L.,Lactuca tatarica (L.)C. A. Meyer.     

Proteins and polypeptides of envelope membranes from spinach chloroplasts. I. Isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis separations

Polypeptides of spinach chloroplast envelopes were separated by electrophoresis in an SDS-polyacrylamide gradient gel. At least 37 polypeptides were resolved; nine were prominent. Two (M_r 54 000 and 16 000) were also found in the stroma fraction and identified by peptide mapping and isoelectric focusing in the second dimension as the large and small subunits of ribulose-1,5-bisphosphate carboxylase. Proteins of the chloroplast envelope were also separated by isoelectric focusing. An adaptation of a previous method (Ames, G.F.L. and Nikaido, K. (1976) Biochemistry 15, 616ndash;623), using solubilization in SDS and isoelectric focusing in the presence of a high concentration of Nonidet P-40, gave the best separation and resolved the envelope membranes into at least 21 proteins. The major band (pI 6.85) contained both subunits of the carboxylase and at least two additional polypeptides which corresponded to the prominent bands found in SDS gel electrophoresis of chloroplast envelopes.