COMPARATIVE MEASUREMENTS OF NUCLEAR DNA IN A HETEROTHALLIC AND A SELF-FERTILE ISOLATE OF THE MYXOMYCETE,DIDYMIUM IRIDIS

Comparative measurements were made of the nuclear Feulgen-DNA content of a heterothallic and a self-fertile isolate of the myxomycete Didymium iridis. Plasmodial nuclei of both isolates contain the diploid amount of DNA. The replicated diploid (4C) values for the heterothallic and the self-fertile isolates are 5.66 and 5.95, respectively. Myxamoebae, however, are quite dissimilar in their nuclear DNA content. Those of the heterothallic isolates, Honduran 1–2 (A¹) and Panamanian 2–4 (A⁷), have mean values of 3.81 and 3.69, whereas myxamoebae of the self-fertile Philippine-1 isolate were found to have a mean value of 6.07. Myxamoebae of the Ph-1 isolate are, therefore, at the same ploidy level as the Ph-1 Plasmodium. Mean DNA values for Ph-1 sporangial nuclei were in category 4C. Measurement of the DNA content of mitotic metaphases in sporangia at T = 6 hr confirmed that the mean DNA content of both Ph-1 myxamoebae and plasmodial nuclei is equivalent to 4C. It is concluded that nuclear phase alternance is lacking in the Ph-1 isolate and that the Plasmodium of this isolate develops by apogamy.     

NUCLEAR DNA CONTENT OF MYXAMOEBAE AND PLASMODIA IN SIX NON-HETEROTHALLIC ISOLATES OF A MYXOMYCETE,DIDYMIUM IRIDIS

The nuclear DNA content of six non-heterothallic isolates of the myxomycete Didymium iridis was measured by combining the Feulgen reaction with absorption microspectrophotometry. This allowed us to distinguish between homothallic (sexual) and apogamic (non-sexual) isolates. Four of the isolates studied, Panamanian 4 and 5, California 1, and Missouri 1 are homothallic. Moreover, the average DNA content of the myxamoebal and plasmodial nuclei (0.32 and 0.61 respectively) does not differ significantly from the calculated haploid and diploid values for heterothallic isolates of D. iridis (0.34 and 0.63). Hence, it is concluded that in each of these isolates the myxamoebae are haploid and the plasmodia diploid. In two of the isolates investigated, Georgia 1 and Hawaii 1, the DNA content of the myxamoebal and plasmodial nuclei did not differ significantly. Therefore, in both of these isolates the plasmodia appear to develop apogamically. In addition the mean DNA values recorded for the Ha-1 isolate suggest that it is aneuploid.     

QUANTITATIVE MICROSPECTROPHOTOMETRY OF NUCLEAR DNA IN SELFING STRAINS OF THE MYXOMYCETE DIDYMIUM IRIDIS

Genetic and cytochemical investigations of the origin, development, nuclear activity, and ploidy level of Plasmodia obtained from selfed clones S-2 and B₁P-33 of the heterothallic myxomycete, Didymium iridis, are presented. To demonstrate that selfing did not result from contamination of the clones, or mutations at the mating-type locus, crosses were made between F₁ clones and clones of known mating types. The data were inconsistent with these two possibilities. DNA was quantified by Feulgen-DNA microspectrophotometry. All cellular phases studied (logarithmic amoebae, swarmers, and encysted amoebae) appear to be haploid, with the nuclear DNA being in the replicated (2C) state. The plasmodia are in all cases diploid; however, the data indicate that the selfed Plasmodia are in an extended G₁ condition. The nuclear DNA content of these is therefore 2C, whereas that of the cross Plasmodium is 4C. Sporangial nuclei exhibit DNA in diploid replicated (4C) category.     

GENETICS OF SOMATIC CELL FUSION IN TWO ISOLATES OF DIDYMIUM IRIDIS

Analysis of progeny from a cross between a Honduran clone and a Panamanian clone of Didymium iridis has revealed existence of five new plasmodial fusion loci in the Panamanian isolate. These are in addition to six which were previously known.     

FURTHER STUDIES IN MULTIPLE ALLELOMORPH HETEROTHALLISM IN THE MYXOMYCETE DIDYMIUM IRIDIS

Analysis of an isolate of Didymium iridis from Costa Rica indicates that it possesses a pair of incompatibility factors. These factors have been shown to be allelic with those of a Honduras and a Panama isolate. Data presented in this paper show that, among them, the 3 isolates possess 6 incompatibility alleles at 1 locus.     

MULTIPLE ALLELES AT THE INCOMPATIBILITY LOCUS IN THE MYXOMYCETE DIDYMIUM IRIDIS

Collins , O'Neil Ray . (Queens Coll., New York City.) Multiple alleles at the incompatibility locus in the Myxomycete Didymium iridis. Amer. Jour. Bot. 50(5): 477–480. 1963.—Working with 2 isolates of Didymium iridis, from Honduras and Panama, respectively, it was shown that each isolate possesses a pair of incompatibility factors. On the basis of results from inter-isolate crosses, one pair was designated A¹ A² and the other A³ A⁴ to indicate that these factors actually constitute a series of multiple alleles at a single locus.     

HETEROTHALLISM AND HOMOTHALLISM: A STUDY OF 27 ISOLATES OF DIDYMIUM IRIDIS,A TRUE SLIME MOLD

Single pore studies of 27 isolates of Didymium iridis from several geographical locations revealed that 11 are heterothallic, displaying a one-locus, multiple allelic mating system. Collectively, 9 of the 11 possess a total of 12 mating types in one multiple allelic series. Each of the remaining two isolates has a pair of mating types which are either incompatible with the 12 or they have not yet been tested against them. Sixteen isolates proved to be homothallic.     

CONTROL OF PLASMODIAL FUSION IN A PANAMANIAN ISOLATE OF DIDYMIUM IRIDIS

Plasmodial fusion in a Panamanian isolate (Pan. 1) of Didymium iridis is controlled by at least six fusion loci. Only plasmodia with identical phenotypes for these six loci are capable of fusion. No evidence was obtained in this study to indicate that any one fusion locus more strongly affects the fusion process than any of the other loci. Control of plasmodial fusion in the Pan. 1 strain of D. iridis is similar to that found earlier for a Honduran strain (Hon. 1).     

GENETICAL AND CYTOLOGICAL EVIDENCE FOR CHROMOSOMAL ELIMINATION IN A TRUE SLIME MOLD,DIDYMIUM IRIDIS

In crosses involving a polyploid myxamoebal clone. CR 2-25*, F₁ plasmodia and myxamoebae display a variety of unexpected ploidy levels as indicated by nuclear DNA measurements. Genetic analyses of the F₁ generations reveal either complete elimination of certain genetic markers or greatly skewed segregation ratios. On the basis of these two kinds of evidence, it is assumed that chromosome elimination occurs at some stage (or stages) following karyogamy between parental nuclei. The possible significance of polyploidy in relation to myxomycete speciation and evolution is discussed.     

SOMATIC CELL INCOMPATIBILITY IN DIDYMIUM IRIDIS: LOCUS IDENTIFICATION AND FUNCTION

Somatic incompatibility in two strains of the myxomycete Didymium iridis is controlled by at least 13 loci: seven fusion loci and six clear-zone loci. Details on correlating loci of the Hon 1 strain with Pan 1 loci are given and a unified nomenclature, applicable to both strains, has been developed from data presented in this paper. Although fusion loci generally prevent fusion between different plasmodial incompatibility phenotypes, studies on individual loci have shown that a limited transient fusion may occasionally take place. Thus, the differences between the fusion and clear-zone loci are not as distinct as once thought. However, the 13 somatic incompatibility loci are still easily designated as either fusion or clear-zone loci, and no locus has thus far been found with true intermediate function. Studies on individual locus function are also discussed.     

NUCLEAR DNA CONTENT IN POPULATIONS OF PINUS RIGIDA

Five populations of Pinus rigida growing in contrasting ecological situations ranging from North Carolina (35°53'N latitude) to Quebec (45°06'N latitude) showed no significant variation in amounts of nuclear DNA with respect to germinating seedlings. Nuclear volume of dormant nuclei also showed no significant variation between and within populations, a finding that is consistent with the concept that the basic 2C DNA value of Pinus rigida is uniform under all habitats. This finding is in contrast to numerous reports for other coniferous species.     

[6N]METHYL ADENINE IN THE NUCLEAR DNA OF A EUCARYOTE, TETRAHYMENA PYRIFORMIS

DNA isolated from macronuclei of the ciliate, Tetrahymena pyriformis, has been found to contain [⁶N]methyl adenine (MeAde); this represents the first clear demonstration of significant amounts of MeAde in the DNA of a eucaryote. The amounts of macronuclear MeAde differed slightly between different strains of Tetrahymena, with approximately 0.65–0.80% of the adenine bases being methylated. The MeAde content of macronuclear DNA did not seem to vary in different physiological states. The level of MeAde in DNA isolated from micronuclei, on the other hand, was quite low (at least tenfold lower than in macronuclear DNA).     

CYTOPHOTOMETRIC DETERMINATION OF THE AMOUNT OF DNA IN ARACHIS L. SECT. ARACHIS (LEGUMINOSAE)

The 2C amounts of DNA for 12 taxa of the section Arachis nom. nud. of the genus Arachis L. were determined using cytophotometric techniques. The diploid taxa ranged from 4.92 to 5.98 pg of DNA per cell. The species of the diploid series Annuae Krap. & Greg. nom. nud. averaged ca. 1 pg less DNA per cell than the taxa of the diploid series Perennes Krap. & Greg. nom. nud. No significant differences were found between taxa within these two series. The tetraploid taxa ranged from 10.36 to 11.35 pg of DNA per cell. Within the tetraploid series Amphiploides Krap. & Greg. nom. nud. differences were found between A. monticola Krap. & Rig. and A. hypogaea L. The two subspecies of A. hypogaea, ssp. hypogaea and ssp. fastigiata Waldron, were found to differ significantly in their 2C amounts of DNA. The implications of the cytophotometric data on the chromosomal evolution of this section are discussed.     

GROWTH AND DEVELOPMENT OF THE MYXOMYCETE PERICHAENA VERMICULARIS. II. CHROMOSOME NUMBERS AND NUCLEAR CYCLES

Chromosome counts of dividing nuclei of the myxomycete Perichaena vermicularis indicate a number of 25 ± 2 in the amoebae and 50 ± 4 in the Plasmodia, confirming earlier reports that amoebae are haploid and plasmodia diploid. Chromosome numbers obtained from nuclei during sporangial development indicate a fluctuation in the location of meiosis influenced by environmental conditions. The implications of these observations are discussed in reference to past conflicting evidence of the location of meiosis.     

FLOW-CYTOMETRIC ANALYSES OF NUCLEAR DNA CONTENT IN FOUR FAMILIES OF NEOTROPICAL BATS

Flow-cytometric analyses of 29 species of microchiropteran bats representing four families and 20 genera revealed that bats possess only 79% (5.43 pg) of the DNA content of a “typical” mammal (e.g., Mus musculus strain C57BL; 7 pg). Chiroptera, the second largest order of mammals, is thus an exception to the prevailing view that mammals possess a minimum nuclear DNA content of 7 pg. Limitations on cell size resulting from a high metabolic rate may have constrained evolution of DNA content and could explain why the extensive heterochromatic additions that are common in some groups of mammals are absent in bats. Chromosomes of bats have been well studied; detailed chromosomal banding data are available for nearly all the species used in this investigation. However, no significant correlations were found between DNA content and karyotypic characteristics such as 2n, fundamental number, and rate or pattern of chromosomal evolution.     

A CYTOPHOTOMETRIC STUDY OF NUCLEIC ACIDS AND PROTEINS IN THE SHOOT APEX OF WHITE SPRUCE

During the annual three-phase growth cycle of white spruce [Picea glauca (Moench) Voss] the vegetative shoot apex changed in anatomical configuration. Relative amounts of DNA, histones, RNA, and proteins were measured in three cytohistological zones and were related to the anatomical changes during ontogeny. An extended period of DNA synthesis (S) and G₂ preceded an increase in the number of apical initial cells which were part of the mammillary apex. While DNA and histones were generally synchronous during ontogeny, the ratio of DNA to histone increased on June 20. This loss of histone and subsequent increases in RNA and cytoplasmic proteins preceded the appearance of needle primordia on next year's apex. We propose that induction of the apex to reorganize and form needle primordia occurred when the DNA was in a 2C condition, following the loss of histone on June 20.     

木耳属真菌rDNA特异性扩增片段的RFLP研究

对木耳属8个种25个菌株的ITS和28SrDNA5’端两个区域分别进行了PCR扩增和限制酶切片段长度多态性（RFLP）研究。ITS－RFMP研究结果表明,HaeⅢ可将黑木耳与其它种区分开,MspⅠ可将盾形木耳、角质木耳、琥珀木耳和黑木耳4个种区分开,而供试的HaeⅢ、TaqⅠ、HinfⅠ和MaPⅠ这四种限制酶均不能将皱木耳、大木耳、网脉木耳及毛木耳4个种区分开,表明它们之间的亲缘关系较近;结果还表明,ITS—rDNA拷贝在毛木耳和琥珀木耳种内是异质性的,而在黑木耳种内是同质性的。285rDNA-RFLP研究结果表明,供试的4种限制酶中,仅MspⅠ可将盾形木耳和角质木耳区分开,而不能将其它种区分开,这显示了28SrDNA序列在木耳属不同种间的保守性。     

DROUGHT STRESS RESPONSES OF MICROSERIS SPECIES DIFFERING IN NUCLEAR DNA CONTENT

Anatomical and physiological responses to drought stress were compared in two Microseris species differing in DNA content and originating from contrasting habitats relative to water availability (M. bigelovii, DNA = 2.6 pg nucleus^–1, more xeric; M. laciniata, DNA = 6.8 pg nucleus^–1, mesic). Leaf mesophyll cell volume was positively correlated with DNA content and negatively correlated with tissue elasticity, i.e., low ϵ̄ and thin cell walls. Drought stress increased leaf tissue elasticity (lower ϵ̄, thinner cell walls). Cell volume, cell wall thickness, cell number, and leaf area were decreased most by drought stress in M. laciniata. Osmotic adjustment with a 20% increase in total solutes (mostly amino acids) after stress was observed in both species, but their estimated contribution to the change in osmotic potential was larger in M. bigelovii. These findings indicate that the Microseris species studied respond to low water availability by maintaining turgor with 1) small cell volumes, 2) elastic tissues (low ϵ̄, thin cell walls), and 3) osmotic adjustment. Both enhanced tissue elasticity and small cell volume appear to be inherent characteristics in M. bigelovii and drought-induced responses in M. laciniata. These data are compatible with the hypothesis that natural selection may influence DNA content through differential sensitivity of cell growth to environmental stress.     

VARIATION OF NUCLEAR DNA CONTENT IN HELIANTHUS ANNUUS (ASTERACEAE)

Nuclear 2C DNA content was determined by laser flow cytometry for 13 diploid (2n = 34) lines (cultivated varieties and inbred lines) of Helianthus annuus. Mean DNA amount of second leaf nuclei varied from 6.01 to 7.95 pg (32%) among lines. Mean DNA content varied up to 19% within lines. Variability in mean DNA content exceeding 27% and 48% was detected among leaves from different nodes of plants of the open-pollinated variety, Californicus, and the inbred line, RHA 299, respectively. The root tip and shoot tip nuclei of H. annuus have been reported to consist of a mixture of aneuploid (17 to 33 chromosomes) and diploid (34 chromosomes) cells, a condition called aneusomaty. Chromosome counts of root tips and an analysis of the distribution of DNA content of large numbers of nuclei from leaves indicate that aneusomaty either does not occur, or is not common, among the lines investigated. The intraspecific, intraline, and intraplant variation in DNA content in H. annuus support the concept that a sizable portion of a plant genome is unstable and subject to rapid changes in DNA amount.