Inheritance of mitochondrial DNA in lentil (Lens culinaris Medik.)

Restriction fragment analysis was used to examine the inheritance of lentil mitochondrial DNA (mtDNA) in F₁ and F₅ progeny from intrasubspecific (Lens culinaris ssp. culinaris) crosses and in F₁ progeny from intersubspecific (Lens culinaris ssp. orientalis x L. culinaris ssp. culinaris) crosses. Southern blots of digested parental and progeny DNA were hybridized to heterologous maize mtDNA probes specific to coxI and atp6 genes. Two restriction fragment polymorphisms separated L.c. ssp. culinaris Laird and Eston from L.c. ssp. culinaris ILL5588, and one restriction fragment polymorphism distinguished L.c. ssp. culinaris Laird and Eston from L.c. ssp. orientalis LO4. Twelve of 13 f₁ progeny and all F₅ progeny from the intrasubspecific crosses, and all F₁ progeny from intersubspecific crosses had only maternal mtDNA restriction fragments. One f₁ plant from an Eston x ILL5588 cross inherited mtDNA fragments from both parents. Nuclear DNA inheritance was biparental in all F₁ progeny.NRCC No. 38451     

Conversion of DDT to DDD in flooded soil

Summary Anaerobic conditions obtained by flooding the soil caused reductive dechlorination of p,p-DDT and its conversion to p,p-DDD was enhanced under water-logged conditions creating or favouring anaerobiosis. The DDT showed recalcitrance in the soil kept at 15% moisture.More o,p-DDT was lost from the flooded soil. Similar amounts of p,p-DDE were detected in all of the three levels of technical DDT treatments and the concentrations were not significantly different under both aerobic and anaerobic conditions.     

DNA/DNA hybridization studies of the carnivorous marsupials. I: The intergeneric relationships of bandicoots (Marsupialia: Perameloidea)

Summary A complete suite of comparisons among six bandicoot species and one outgroup marsupial was generated using the hydroxyapatite chromatography method of DNA/DNA hybridization; heterologous comparisons were also made with three other bandicoot taxa. Matrices of T_m's, modes, and T₅₀Hs were generated and corrected for nonreciprocity, homoplasy, and, in the case of T_m's, normalized percent hybridization; these matrices were analyzed using the FITCH algorithm in Felsenstein's PHYLIP (version 3.1). Uncorrected and nonreciprocity-corrected matrices were also jackknifed and analyzed with FITCH to test for consistency. Finally, sample scores for T_m, mode, and T₅₀H matrices were bootstrapped and then subjected to phylogenetic analysis. These manipulations were carried out, in part, to address criticisms of the statistics used to summarize DNA/DNA hybridization (especially T₅₀H) and the method itself. However, with the exception of an unresolved trichotomy among the twoEchymipera species andPeroryctes longicauda, all trees showed the same branchpoints. Except in the case of the tree generated from reciprocal-corrected T_m data, nodes were stable under jackknifing; and, again excepting the above-mentioned trichotomy, all nodes were supported by 95% or more of the bootstrapped trees. These results suggest that, despite arguments to the contrary, all three summary statistics can be valid for DNA/DNA hybridization data. Of taxonomic interest is the placement ofEchymipera spp. andPeroryctes longicauda together and separate from the more distantPeroryctes raffrayanus; the genusPeroryctes is thus at least paraphyletic. The trees further groupedEchymipera-plus-Peroryctes as the sister group ofIsoodon-plus-Perameles. Limited hybridizations withMacrotis lagotis suggest that its current position as representative of an entirely distinct family of perameloids is correct.This article was presented at the C.S.E.O.L. Conference on DNA-DNA Hybridization and Evolution, Lake Arrowhead, California, May 11–14, 1989     

Nor loci analysis in progenies of plants regenerated from the scutellar callus of bread-wheat

Summary Progenies of plants regenerated from scutellar callus of bread wheat (Triticum aestivum L.) were analysed for the organization of the intergenic spacer of the rRNA genes, located at the sites of the nucleolar organizer region (Nor loci). Sexual progenies derived from the regenerated plants of three wheat cultivars were subjected to this analysis. The respective DNAs were digested with the restriction endonuclease TaqI, and probed with a specific rDNA fragment by Southern blothybridization. The intergenic rDNA spacer could thus be characterized for each of the three cultivars. Thirty-eight progeny plants of the cultivars Chinese Spring and Miriam were found to be stable in their organisation of the Nor loci: no changes relative to the Nor of control plants from these cultivars were revealed. On the other hand, three progeny plants of ND7532 showed reduction in the number of the rDNA spacers. Since no variability in the Nor loci could be revealed among control ND7532 plants, this seems to indicate that the changes in the progeny of regenerated plants resulted from the in vitro culture of the scutellar callus. Grain glutenin and gliadin profiles of sexual progenyplants derived from scutellar calli of Chinese Spring, of Miriam as well as of ND7532 were identical to the respective control plants of these cultivars, indicating low (or no) somaclonal variation in these grain proteins in the analyzed plants.     

The relationship between Photosystem II intrinsic quantum yield and millisecond luminescence in thylakoids

The relationship between charge recombination at Photosystem II (PS II), as indicated by millisecond luminescence, and PS II quantum yield was studied in spinach thylakoids during electron flow to methylviologen. Under the low magnesium conditions used, a decrease in quantum yield was observed in the absence of non-photochemical excitation quenching, and therefore cannot be due to a restriction in excitation delivery to the reaction centre. It was found that the decrease of the parameter _p, which is a measure of the intrinsic quantum yield of open PS II centers, correlates with an increase in luminescence per open center. The relationship between these two parameters was the same whether _p was manipulated by dissipation of the transthylakoid pH gradient or of the electrical potential. This indicates that the mechanism by which _p decreases depends in the same way on the two components of the protonmotive force as does the charge recombination at PS II. Calculation of the yield of luminescence with respect to the back reaction will be necessary to determine whether the charge recombination occurs at a sufficiently high rate to be directly responsible for the _p decrease.     

Genotype-independent leaf disc transformation of potato (Solanum tuberosum) using Agrobacterium tumefaciens

Summary Leaves of the in vitro grown potato cultivars Bintje, Berolina, Desiree, and Russet Burbank were wounded and co-cultivated with Agrobacterium strains having chimeric bar and nptII genes on a disarmed T-DNA. Each leaf from these cultivars formed numerous calli on kanamycin-containing medium, and almost all calli regenerated shoots. For Russet Burbank, it was necessary to include AgNO₃ in the medium to obtain efficient shoot regeneration. The transformed plants have one to a few copies of the T-DNA, show NPT-II and PAT activities, and are resistant to high doses of the commercial preparation of phospinotricin (glufosinate). Almost no somaclonal variation was detected in trans-genic plants.     

Molecular mapping of a new gene Rrs4 CI 11549 for resistance to barley scald (Rhynchosporium secalis)

Doubled haploid (DH) progeny from a cross between the scald susceptible barley (Hordeum vulgare L.) cultivar Ingrid and the resistant accession CI 11549 (Nigrinudum) was evaluated for resistance in the pathogen Rhynchosporium secalis (Oudem) J.J. Davis. Two linked and incompletely dominant loci confer resistance CI 11549 against isolate 4004. One is an allele at the complex Rrs1 locus on chromosome 3H close to the centromere; the other is located 22 cM distally on the long arm. The latter locus is designated Rrs4. In BC₃-lines into Ingrid from CI 2222 (another Nigrinudum) resistance seems governed by one locus close to the telomeric region of chromosome 7H, probably allelic to Rrs2. In neither case did we find any trace of the recessive gene rh8 reported to be present in Nigrinudum. Various resistance donors of Ethiopian origin designated as Nigrinudum, Jet or Abyssinian were identical to a great extent with respect to markers, but differed in resistance to different isolates of scald or in barley yellow dwarf virus (BYDV) resistance. The implications for their use as differentials in scald tests and screening of germplasm collections are discussed.     

The origin of Q-independent derivatives of phage lambda

Summary qsr (Q-independent) phages are characterised by the replacement of the region of the genome that contains Q, S, R, and the late gene promoter, P_R, with host-derived DNA that codes for functions analogous to those deleted. Restriction endonuclease analysis and DNA/DNA hybridisation methods have been used to show that p4 and qin A 3, two such Q-independent phages, are the product of recombination between and a defective lambdoid prophage (the qsr prophage) located at an as yet unidentified site in the E. coli K 12 chromosome. The qsr prophage is distinct from the defective lambdoid prophage Rac (Kaiser and Murray 1979). In the E. coli K 12 strain AB1157 from which qsr phages cannot be generated, the qsr prophage has suffered an internal deletion. That the qsr prophage appears not to carry a full complement of essential late genes suggests one explanation for its apparently defective nature.     

The ability of acidic pH,growth inhibitors,and glucose to increase the proton motive force and energy spilling of amino acid-fermenting <Emphasis Type="Italic">Clostridium sporogenes</Emphasis> MD1 cultures

Clostridium sporogenes MD1 grew rapidly with peptides and amino acids as an energy source at pH 6.7. However, the proton motive force (p) was only –25 mV, and protonophores did not inhibit growth. When extracellular pH was decreased with HCl, the chemical gradient of protons (ZpH) and the electrical membrane potential () increased. The p was –125 mV at pH 4.7, even though growth was not observed. At pH 6.7, glucose addition did not cause an increase in growth rate, but increased to –70 mV. Protein synthesis inhibitors also significantly increased . Non-growing, arginine-energized cells had a of –80 mV at pH 6.7 or pH 4.7, but was not detected if the F₁F₀ ATPase was inhibited. Arginine-energized cells initiated growth if other amino acids were added at pH 6.7, and and ATP declined. At pH 4.7, ATP production remained high. However, growth could not be initiated, and neither nor the intracellular ATP concentration declined. Based on these results, it appears that C. sporogenes MD1 does not need a large p to grow, and p appears to serve as a mechanism of ATP dissipation or energy spilling.Mandatory disclaimer: Proprietary or brand names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by the USDA implies no approval of the product, and exclusion of others that may be suitable.     

Exposure to Orange (Citrus sinensis L.) Trees, Fruit, and Oil Enhances Mating Success of Male Mediterranean Fruit Flies (Ceratitis capitata [Wiedemann])

Previous laboratory tests revealed that exposure to oranges (Citrus sinensis L.) increased the mating success of male Mediterranean fruit flies, Ceratitis capitata (Wiedemann) (medfly). This advantage may have resulted from male exposure to -copaene (a sesquiterpene hydrocarbon and known male attractant) in the peel, as pure -copaene has been shown to increase the mating success of male medflies. Working with orange trees as well, we investigated whether male exposure to nonfruiting trees, leaves (also known to contain -copaene albeit at a lower concentration than fruit), and fruit conferred a mating advantage to wild-like males in field-cage tests. Males exposed to entire nonfruiting trees or leaves had a mating advantage over control males (exposed to a nonhost plant) in trials conducted 1 day but not 3 days after exposure. Males exposed to orange fruits had higher mating success than control males (exposed to apples) in trials conducted 1 and 3 days after exposure. Enhanced mating success was observed only when males were permitted to contact the orange leaves and fruits; aroma alone did not affect male mating success. In addition, we examined whether exposure to commercially available orange oil, which also contains -copaene, enhanced the mating performance of wild-like and mass-reared sterile males. Heightened mating success was observed in trials conducted 1 and 3 days after exposure for both types of males, and in this case aroma alone had a positive effect on male mating success. Future research should attempt to identify the behavioral, physiological, or chemical mechanisms underlying the observed increases in male mating success.     

The catalysis of nucleotide polymerization by compounds of divalent lead

Summary Soluble lead salts and a number of lead-containing minerals catalyze the formation of oligonucleotides from nucleoside 5-phosphorimidazolides. The effectiveness of lead compounds correlates strongly with their solubility. Under optimal conditions we were able to obtain 18% of pentamer and higher oligomers from ImpA. Reactions involving ImpU gave smaller yields.Abbreviations A adenosine - U uridine - Im imidazole - MeIm 1-methyl-imidazole - EDTA ethylenediaminetetraacetic acid - pA adenosine 5-phosphate - pU uridine 5-phosphate - Ap adenosine cyclic 2:3-phosphate - ATP adenosine 5-triphosphate - AppA P₁,P₂-diadenosine 5-diphosphate - pNp (N = A,U) nucleotide 2(3), 5-diphosphate - ImpA adenosine 5-phosphoreimidazolide - ImpU uridine 5-phosphorimidazolide - A ²pA adenylyl-[25]-adenosine - A ³pA adenylyl-[35]-adenosine - pA ²pA 5-phospho-adenylyl-[25]-adenosine - pA ³pA 5-phospho-adenylyl-[35]-adenosine - pUpU 5-phospho-uridylyl-uridine - pApU 5-phospho-adenylyl-uridine - pUpA 5-phospho-uridylyladenine - (pA)n (n, 2,3,4,) oligoadenylates with 5 terminal phosphate - ImpApA 5-phosphorimidazolide of adenylyl adenosine - (pA) ₅₊ pentamer and higher oligoadenylates with 5 terminal phosphate - (Ap)nA (n = 2,3,4) oligoadenylates without terminal phosphates In the following we do not specify the nature of the internucleotide linkageIn the following we do not specify the nature of the internucleotide linkage     

Immature embryo rescue,culture and seedling development of acid citrus fruit derived from interploid hybridization

Interploid sexual hybridizations were completed in 2001 and 2002 between seven lemon (Citrus limon(L.) Burm. f.) varieties, Key lime (C. aurantifolia (Cristm.) Swing), Palestine sweet lime (C. imettioides Tan.), Lakeland limequat (C. aurantifolia x Fortunella japonica (Thumb.) Swing.), and Etrog citron (C. medica L.) as diploid progenitors and four allotetraploid somatic hybrids (Key lime + Valencia orange, Hamlin orange + Femminello lemon, Valencia orange + Rough lemon, and Valencia orange+ Femminello lemon) in efforts to generate improved seedless triploid acid fruit hybrids. Efficient recovery of triploid progeny from such crosses requires embryo rescue to avoid embryo abortion due to endosperm failure. Germination of rescued genetically diverse immature embryos was induced on two culture media (EME and Gamborgs B5), with two sucrose concentrations (50 or 70 g l^–1). All media contained 0.5 g l^–1 malt extract and 4.50 M GA₃. Germination of globular, heart and torpedo shaped embryos (defined as small embryos) was significantly (p < 0.05) affected by medium and genotype. Gamborgs medium induced 82.89% germination. Of germinated embryos, 11–65% developed into normal plants with differences among crosses. Cotyledonary embryos (defined as immature embryos with fully developed cotyledons) germinated and developed into normal plants at higher rates than less-developed embryos. In efforts to improve the efficiency of plant recovery, small embryos from Todo el año × HF and Lisbon × HF crosses conducted during 2002 were rescued and cultured on three media (MS, Gamborgs, and RMA) for comparison. Media did not significantly affect the proportion of normal plant recovery.     

Use of RFLP markers for the identification of alleles of the Pm3 locus conferring powdery mildew resistance in wheat (Triticum aestivum L.)

The objective of this study was to identify molecular markers linked to genes for resistance to powdery mildew (Pm) in wheat using a series of Chancellor near-isogenic-lines (NILs), each having one powdery mildew resistance gene. A total of 210 probes were screened for their ability to detect polymorphism between the NILs and the recurrent parent. One of these restriction fragment length polymorphism (RFLP) markers (Xwhs179) revealed polymorphism not only between the NILs for the Pm3 locus, but also among NILs possessing different alleles of the Pm3 locus. The location of the marker Xwhs179 was confirmed to be on homoeologous chromosome group 1 with the help of nullitetrasomic wheat lines. The linkage relationship between this probe and the Pm3 locus was estimated with double haploid lines derived from a cross between wheat cvs Club and Chul (Pm3b). The genetic distance was determined to be 3.3±1.9 cM.     

Effects of soil pH and associated cations on growth of apple trees planted in old orchard soil

Summary Trunk circumferences were measured on apple trees which had been grown for 5 to 10 years on land previously in apple orchard. Soil beneath each tree was analysed for soil acidity characteristics and basic cations. Tree growth (trunk circumference), was generally retarded but highly variable, but correlated well with the soil measurements in four of the six orchards investigated. Size of Delicious, Tydeman and Rome Beauty cultivars correlated positively with soil pH and negatively with 0.02M CaCl₂-soluble Al and Mn, while that of McIntosh grown adjacent to the Delicious and Tydeman trees in two of those orchards correlated little or not at all with these soil acidity characteristics. However, in the three orchards where McIntosh trees were grown, their size correlated (r=0.72^**, 0.71^**, 0.54^**) well with exchangeable soil Mg. The effects of soil acidity and associated nutrient deficiencies on growth of young apple trees is discussed.Contribution No. 602.     

Transfer of Ogu cytoplasmic male sterility to Brassica juncea and improvement of the male sterile line through somatic cell fusion

Male sterility conferred by ogu cytoplasm of Raphanus sativus has been transferred to Brassica juncea cv RLM 198 from male-sterile B. napus through repeated backcrossing and selection. The male-sterile B. juncea is, however, highly chlorotic and late. It has low female (seed) fertility and small contorted pods. To rectify these defects, protoplasts of the male sterile were fused with normal RLM 198 (green, self fertile). Four dark green, completely male-sterile plants were obtained and identified as putative cybrids. All the plants were backcrossed three times with RLM 198. Mitochondrial and chloroplast DNA analysis of backcross progeny confirmed hybridity of the cytoplasm. The restriction pattern of the chloroplast DNA of progeny plants of three cybrids (Og 1, Og 2, Og 3) was similar to that of the green self-fertile RLM 198 and indicated that the correction of chlorosis resulted from chloroplast substitution. The chloroplast DNA of the lone progeny plant of the fourth cybrid (Og 10) could not be analyzed because the plant was stunted and had only a few leaves. When total cellular DNA was probed with mitochondrial probes coxI and atpA it was found that the cybrids had recombinant mitochondria. The chlorosis-corrected plants were early flowering and had vastly improved seed fertility.     

Mg2+ Induces Conformational Changes in the Catalytic Subunit of Phosphorylase Kinase, Whether by Itself or as Part of the Holoenzyme Complex

Phosphorylase kinase (PhK) from skeletal muscle is a structurally complex, highly regulated, hexadecameric enzyme of subunit composition ()₄. Previous studies have revealed that the activity of its catalytic subunit is controlled by alterations in quaternary structure initiated at allosteric and covalent modification sites on PhK's three regulatory subunits; however, changes in the conformation of the holoenzyme initiated by the catalytic subunit have been more difficult to document. In this study a monoclonal antibody (mAb 79) has been generated against isolated subunit and used as a conformational probe of that subunit. The epitope recognized by this antibody is within the catalytic core of the subunit, between residues 100 and 240, and monovalent fragments of the antibody inhibit the catalytic activity of the holoenzyme, the -calmodulin binary complex, and the free subunit. Activation of PhK by a variety of mechanisms known or thought to act through its regulatory subunits (phosphorylation, ADP binding, or alkaline pH) increased the binding of the holoenzyme to immobilized mAb 79, indicating that activation by any of these distinct mechanisms involves repositioning of the portion of the catalytic domain of the subunit containing the epitope for mAb 79. The activating ligand Mg²⁺ also stimulated the binding of the PhK holoenzyme to immobilized mAb 79, as well as the binding of mAb 79 to immobilized subunit. Thus, Mg²⁺ increases the accessibility of the mAb 79 epitope in both the isolated subunit and in the holoenzyme. Our results suggest that previously reported influences of Mg²⁺ on the quaternary structure of the PhK holoenzyme are directly mediated by the subunit.     

Aberrations in sexual behavior in some brewing and other industrial yeasts

Summary The mating behavior of a number of brewer's and distiller's yeasts was determined with a and haploid and aa and diploid tester strains. Mating frequencies were not high, ranging from one to (rarely) 2,000/10⁸ cells in the mating mixture. Sporulating hybrids were obtained in most matings, though the percentage spore viability initially obtained was often low. Notable the spore viability obtained in hybrids with the haploid tester strains and the brewing strains DIB and DICH was much higher than from the a haploid tester strain, and higher in hybrids between these strains and the aa diploid tester than in those from the tester strain. With the brewing strain NBA, the spore viability in hybrids with the a haploid tester strain was higher than in the case of strains DIB and DICH, but the spore viability in the hybrid of NBA x the haploid strain was higher still. The data are consistent with the hypothesis that with the a and aa tester strains, most of the industrial yeasts tested mate as diploids, and with the and testers, they mate as haploids, an hypothesis which is supported by the segregation of adenine markers in the progeny of these hybrids.Presented in part at the 6th International Specialized Symposium on Yeasts, Montpellier, France, 2–8 July, 1978     

Combined effect of cytochalasin a and adenosine triphosphate onPhysarum polycephalum

Summary Physarum polycephalum microplasmodia exposed to 1.6×10^–5 M cytochalasin A evidenced intracellular cytoplasmic condensation, slow contraction, and eventual breaks at discrete surface areas, within one hour. Other cytochalasins tested (CB or CD) did not substitute for CA. CA effects on plasmodia were not abolished by immediate washing or media replacement. In nutrient medium, CA plus ATP (375 M) produced within minutes herniation (blebbing) and plasmodial disruption. The order of addition of reagents was important; ATP added simultaneously with or prior to CA stimulated the phenomenon, whereas initial addition of CA resulted in no such dynamic response. Several other nucleotides (e.g., AMP, cAMP) could substitute for ATP; however, such changes were not observed with 5-adenylylimidodiphosphate. Blebbing was not abolished in the presence of 2,4-dinitrophenol. In minimal medium, it was best stimulated by simultaneous addition of Ca⁺⁺ and Mg⁺⁺. Preincubation of CA with L-cysteine or with -mercaptoethanol negates its individual or nucleotide-combined effects. Yet, 10^–5 M ethacrynic acid, a sulfhydryl-reactive liposoluble drug, in the presence of ATP does not mimic the blebbing response. These observed effects, which take place at or near the plasmodial surface, presumably reflect acceleration of normal contractile processes inPhysarum. Abbreviations CA cytochalasin A - CB cytochalasin B - CD cytochalasin D - AMP adenosine 5-monophosphate - ADP adenosine 5-diphosphate - ATP adenosine 5-triphosphate - di-butyryl-cAMP di-butyryl-cyclic adenosine 35-monophosphate - di-butyrylcGMP di-butyryl-cyclic guanosine 35-monophasphate. This work was supported by a grant (AI-11902) from the U.S. Public Health Service.     

Molecular systematics and genetic differentiation of Pinus sylvestris (L.) and P. densiflora (Sieb. et Zucc.)

Summary Allozyme variation was examined in 22 populations of Pinus densiflora (Sieb, et Zucc.) and four geographic varieties of P. sylvestris (L.): var lapponica (Fries, Hartman), var armena (Komarov), var mongolica (Litvinov) and var sylvestriformis (Takenouchi). In addition, we developed paternal chloroplast (cp) DNA markers that distinguish P. densiflora from var lapponica, var armena and var mongolica. UPGMA cluster analysis based on Nei's distances between all pairwise combinations of the 22 populations revealed patterns corresponding strictly to geographic origin and taxonomic status. Analysis of allozyme variation in var lapponica, var armena and var mongolica demonstrated a high level of intrapopulational variability but a low level of interpopulational differentiation. It appears that the late Pleistocene blending of genetically diverse populations was responsible for the observed variation patterns. The constructed phylogenetic trees also showed late divergence of these three varieties. The var sylvestri formis was genetically distinct from the other three P. sylvestris varieties. The genetic distances separating var sylvestriformis from P. densiflora and the other taxa lend support to a separate taxonomic status for var sylvestriformis and a close relation with P. densiflora. We found that var sylvestriformis harbors admixtures of allozymes and cpDNA from both P. sylvestris and P. densiflora, which suggests an introgressive nature of this variety. Levels of intrapopulational variability were similar in P. sylvestris and P. densiflora, but interpopulational differentiation was much higher in P. densiflora. In the constructed phylogenetic trees, populations of this species were characterized by relatively long internode distances and branch lengths. The present results suggest that P. densiflora has a more advanced evolutionary age than P. sylvestris.