Competição entre megaspórios em milho

The experiments reported were started as early as 1933, when indications were found in class material that the factor for small pollen, spl, causes not only differences in the size of pollen grains and in the growth of pollen tubes, but also a competition between megaspores, as first observed by RENNER (1921) in Oenothera. Dr. P. C. MANGELSDORF, who had kindly furnished the original seeds, was informed and the final publication delayed untill his publication in 1940. A further delay was caused by other circunstances. The main reason for the differences of the results obtained by SINGLETON and MANGELSDORF (1940) and those reported here, seems to be the way the material was analysed. I applied methods of a detailed statistical analysis, while MANGELSDORF and SINGLETON analysed pooled data. 1) The data obtained on pollen tube competition indicate .that there is about 3-4% of crossing-over between the su and sp factors in chromosome IV. The elimination is not always complete, but from 0 to 10% of the sp pollen tubes may function, instead of the 50% expected without elimination. These results are, as a whole, in accordance with SINGLETON and MANGELSDORF's data. 2) Female elimination is weaker and transmission determined as between 16 to 49,5%, instead of 50% without competition, the values being calculated by a special formula. 3) The variability of female elimination is partially genotypical, partially phenotypical. The former was shown by the difference in the behavior of the two progenies tested, while the latter was very evident when comparing the upper and lower halves of ears. For some unknown physiological reason, the elimination is generally stronger in the upper than in the lower half of the ear. 4) The female elimination of the sp gene may be caused theoretically, by either of two processes: a simple lethal effect in the female gametophyte or a competition between megaspores. The former would lead not only to the abortion of the individual megaspores, but of the whole uniovulate ovary. In the case of the latter, the abortive megaspore carrying the gene sp will be substituted in each ovule by one of the Sp megaspores and no abortion of ovaries may be observed. My observations are completely in favor of the second explication: a) The ears were as a whole very well filled except for a few incomplete ears which always appear in artificial pollinations. b) Row arrangement was always very regular. c) The number of kernels on ears with elimination is not smaller than in normal ears, but is incidentally higher : with elimnation, in back-crosses 354 kernels and in selfed ears 390 kernels, without elimination 310 kernels per ear. d) There is no correlation between the intensity of elimination and the number of grains in individual ears; the coefficient; of linear correlation, equal to 0,24, is small and insignificant. e) Our results are in complete disagreement whit those reported by SINGLETON and MANGELSDORF (1940). Since these authors present only pooled date, a complete and detailed analysis which may explain the cause of these divergences is impossible.

A ação dos gens gametofíticos com referência especial ao milho

1) The first part deals with the different processes which may complicate Mendelian segregation and which may be classified into three groups, according to BRIEGER (1937b) : a) Instability of genes, b) Abnormal segregation due to distur- bances during the meiotic divisions, c) obscured segregation, after a perfectly normal meiosis, caused by elimination or during the gonophase (gametophyte in higher plants), or during zygophase (sporophyte). Without entering into detail, it is emphasized that all the above mentioned complications in the segregation of some genes may be caused by the action of other genes. Thus in maize, the instability of the Al factor is observed only when the gene dt is presente in the homozygous conditions (RHOADES 1938). In another case, still under observation in Piracicaba, an instability is observed in Mirabilis with regard to two pairs of alleles both controlling flower color. Several cases are known, especially in corn, where recessive genes, when homozigous, affect the course of meiosis, causing asynapsis (asyndesis) (BEADLE AND MC CLINTOCK 1928, BEADLE 1930), sticky chromosomes (BEADLE 1932), supermunmerary divisions (BEADLE 1931). The most extreme case of an obscured segregatiou is represented by the action of the S factors in self stetrile plants. An additional proof of EAST AND MANGELSDORF (1925) genetic formula of self sterility has been contributed by the studies on Jinked factors in Nicotina (BRIEGER AND MANGELSDORF (1926) and Antirrhinum (BRIEGER 1930, 1935), In cases of a incomplete competition and selection between pollen tubes, studies of linked indicator-genes are indispensable in the genetic analysis, since it is impossible to analyse the factors for gametophyte competition by direct aproach. 2) The flower structure of corn is explained, and stated that the particularites of floral biology make maize an excellent object for the study of gametophyte factors. Since only one pollen tube per ovule may accomplish fertilization, the competition is always extremely strong, as compared with other species possessing multi-ovulate ovaries. The lenght of the silk permitts the study of pollen tube competitions over a varying distance. Finally the genetic analysis of grains characters (endosperm and aleoron) simpliflen the experimental work considerably, by allowing the accumulation of large numbers for statistical treatment. 3) The four methods for analyzing the naturing of pollen tube competition are discussed, following BRIEGER (1930). Of these the first three are: a) polinization with a small number of pollen grains, b) polinization at different times and c) cut- ting the style after the faster tubes have passe dand before the slower tubes have reached the point where the stigma will be cut. d) The fourth method, alteration of the distatice over which competition takes place, has been applied largely in corn. The basic conceptions underlying this process, are illustrated in Fig. 3. While BRINK (1925) and MANGELSDORF (1929) applied pollen at different levels on the silks, the remaining authors (JONES, 1922, MANGELSDORF 1929, BRIEGER, at al. 1938) have used a different process. The pollen was applied as usual, after removing the main part of the silks, but the ears were divided transversally into halves or quarters before counting. The experiments showed generally an increase in the intensity of competition when there was increase of the distance over which they had to travel. Only MANGELSDORF found an interesting exception. When the distance became extreme, the initially slower tubes seemed to become finally the faster ones. 4) Methods of genetic and statistical analysis are discussed, following chiefly BRIEGER (1937a and 1937b). A formula is given to determine the intensity of ellimination in three point experiments. 5) The few facts are cited which give some indication about the physiological mechanism of gametophyte competition. They are four in number a) the growth rate depends-only on the action of gametophyte factors; b) there is an interaction between the conductive tissue of the stigma or style and the pollen tubes, mainly in self-sterile plants; c) after self-pollination necrosis starts in the tissue of the stigma, in some orchids after F. MÜLLER (1867); d) in pollon mixtures there is an inhibitory interaction between two types of pollen and the female tissue; Gossypium according to BALLS (1911), KEARNEY 1923, 1928, KEARNEY AND HARRISON (1924). A more complete discussion is found in BRIEGER 1930). 6) A list of the gametophyte factors so far localized in corn is given. CHROMOSOME IV Ga 1 : MANGELSDORF AND JONES (1925), EMERSON 1934). Ga 4 : BRIEGER (1945b). Sp 1 : MANGELSDORF (1931), SINGLETON AND MANGELSDORF (1940), BRIEGER (1945a). CHROMOSOME V Ga 2 : BRIEGER (1937a). CHROMOSOME VI BRIEGER, TIDBURY AND TSENG (1938) found indications of a gametophyte factor altering the segregation of yellow endosperm y1. CHROMOSOME IX Ga 3 : BRIEGER, TIDBURY AND TSENG (1938). While the competition in these six cases is essentially determined by one pair of factors, the degree of elimination may be variable, as shown for Ga2 (BRIEGER, 1937), for Ga4 (BRIEGER 1945a) and for Spl (SINGLETON AND MANGELSDORF 1940, BRIEGER 1945b). The action of a gametophyte factor altering the segregation of waxy (perhaps Ga3) is increased by the presence of the sul factor which thus acts as a modifier (BRINCK AND BURNHAM 1927). A polyfactorial case of gametophyte competition has been found by JONES (1922) and analysed by DEMEREC (1929) in rice pop corn which rejects the pollen tubes of other types of corn. Preference for selfing or for brothers-sister mating and partial elimination of other pollen tubes has been described by BRIEGER (1936). 7) HARLAND'S (1943) very ingenious idea is discussed to use pollen tube factors in applied genetics in order to build up an obstacle to natural crossing as a consequence of the rapid pollen tube growth after selfing. Unfortunately, HARLAND could not obtain the experimental proof of the praticability of his idea, during his experiments on selection for minor modifiers for pollen tube grouth in cotton. In maize it should be possible to employ gametophyte factors to build up lines with preference for crossing, though the method should hardly be of any practical advantage.

Preferential reproduction mode of hermaphrodite papaya plant (Carica papaya L; Caricaceae)

This research was done to study the reproductive system of papaya hermaphrodite plant based on the histochemical nature of pollen grain, stigma receptivity, in vivo pollen grain germination and pollen:ovule ratio. In the histochemical analysis, pollen grains were stained by using Sudan IV and I2KI solution ; the stigma receptivity was assessed by alpha-naphthtyl acetate solution in closed and opened flowers. Pollen germination and pollen tube growing were examined in flower buds near anthesis with 0.1% aniline blue. To estimate the pollen:ovule ratio , anthers from each flower bud were dissected and all pollen grains were counted; ovules were dissected from ovaries and were counted under stereomicroscope. The results indicated that papaya pollen grains are of lipidic nature; the stigmas were receptive before the opening and until 48 hours after opening; the pollen grains germinated and emitted polinic tube before flower opening and the pollen:ovule ratio indicated the predominance of autogamous reproductive system. These results indicate that hermaphrodite papaya trees is preferentially of optional autogamous with cleistogamy.

Reproductive biology of the protandrous Ferdinandusa speciosa Pohl (Rubiaceae) in Southeastern Brazil

A study of the floral biology and the breeding system of Ferdinandusa speciosa Pohl (Rubiaceae) was carried out from March to September 1996 in Uberlândia, MG, central Brazil. This species is a shrub or small tree that occurs in swampy edges of gallery forests. The two studied populations flowered somewhat asynchronously from March to July. The tubular flowers are red, approximately 4.7 cm long and last for two days. They are protandrous and the pollen is available one day before the stigma becomes receptive. The beginning of anthesis and the opening of the stigmatic lips occur at dusk. The nectar is secreted during both the male and the female phases, with concentration of sugars greater in the male phase. The flowers are pollinated by two hummingbird species, Chlorostilbon aureoventris and Phaethornis pretrei. Ferdinandusa speciosa is a self-compatible, non-apomictic species, with low fruit production under natural conditions in the populations studied. No differences were found between fruit set of self- and cross-pollinated flowers, nor in the pollen tube growth rate in the pistils of these flowers. The seeds formed by cross-pollination are larger, heavier and show a higher percentage of germination than those formed by self-pollination, which indicates inbreeding depression. This result suggests that, although the species is self-compatible, cross-pollination may be advantageous.

Distyly and variation in floral traits in natural populations of Psychotria ipecacuanha (Brot.) Stokes (Rubiaceae)

Psychotria ipecacuanha is a perennial, medicinal herb that grows in clusters in the understory of humid, shady areas of the Atlantic Rain Forest of southeastern Brazil. The present study characterized the variation in floral traits among 35 clusters from three natural populations of this plant species. Field observations showed that the clusters are isomorphic, that is, a given cluster will either set long-styled or short-styled flowers. Stigmas and anthers are reciprocally placed in each morph, a dimorphism characteristic of distyly. The populations are isoplethic, that is, a given population exhibits an equilibrium 1:1 ratio of floral morphs. Morphometric analyses revealed that anther length, stigma length, corolla diameter, and pollen grain diameter were consistently greater in short-styled flowers, regardless of the population investigated. Significant differences for floral traits in the short-styled morph were found among populations. Floral traits in the long-styled morph also showed some significant differences among populations, but not for stigma height and corolla length. Controlled pollinations carried out in natural populations showed that fruit production was higher after inter-morph pollination. Nevertheless, observations of pollen tube growth in style, and also fruit production after spontaneous self-pollination and intra-morph pollination, indicated partial intramorph compatibility in this plant species.

Reproductive biology of the herkogamous vine Chiococca alba (L.) Hitchc. (Rubiaceae) in the Atlantic Rain Forest, SE Brazil

The family Rubiaceae comprises a wide spectrum of floral mechanisms and two of them seem to be common in certain groups, e.g., distyly in Rubioidae and styllar pollen in Ixoroidae. These mechanisms include herkogamy, which is interpreted as a strategy that avoids self-pollination. This is the first report on the reproductive biology of Chiococca alba, a species that is widely distributed in America. We studied floral biology and the mating system, which were evaluated through fruit set comparisons after controlled crosses (self- and cross-pollinations and test for apomixis), as well as through the evaluation of pollen tube growth resulting from these controlled crosses. Flowers of C. alba are herkogamous, cream, protandrous and lasted for two days. No measurable nectar was found, despite the presence of a nectary-like structure at the base of the corolla tube. Chiococca alba is a preferentially self-incompatible species, but self-pollination and apomixis also contribute to the natural fruit-set. Its reproductive strategy (herkogamy associated with protandry) is different from that expected for members of Chiococceae tribe (i.e., styllar pollen presentation).

Pollination biology in Jacaranda copaia (Aubl.) D. Don. (Bignoniaceae) at the "Floresta Nacional do Tapajós", Central Amazon, Brazil

Jacaranda copaia (Aubl.) D. Don is a pioneer tree widespread in the Brazilian Amazon, usually found colonizing forest gaps and altered areas, and the forest fragment edges. This study investigated aspects of the floral biology, breeding system and pollinators of J. copaia trees. Flowering lasts from August to November, during the low rainfall period extending up to four weeks per tree and 3-4 months for the population as a whole, characterizing a cornucopia flowering pattern. The fruit set ends in the beginning of the rainy season, with wind dispersed winged seeds. Fruit set from open pollination was 1.06% (n = 6,932). Hand pollination using self-pollen (n = 2,099) did not set fruits. Cross-pollination resulted in 6.54% fruit set (n = 2,524), representing six times more than the natural pollination rate (1.06%, n = 6,932). Flowers excluded from insect visitation (automatic self-pollination) did not set fruits (n = 5,372). Pollen tube growth down to ovary was detected under fluorescence microcoscopy in cross-pollinated and selfed pistils. The species is an obligate allogamous plant, with late-acting self-incompatibility system. Approximately 40 species of native bees visited the flowers, but the main pollinators were medium-sized solitary bees as Euglossa and Centris species due to the compatibility between their body sizes with the corolla tube, direct contact with the reproductive structures and high frequency of visits.

Reproductive biology of Protium spruceanum (Burseraceae), a dominant dioecious tree in vegetation corridors in Southeastern Brazil

We investigated the reproductive biology of Protium spruceanum (Benth.) Engler in vegetation corridors of secondary Atlantic forest in Lavras, southern Minas Gerais State, Brazil. The reproductive phenology was investigated fortnightly over a one year period. Floral biology studies involved pollen viability analysis, nectar production, stigmatic receptivity, pollen tube growth, visiting insect species and visit rates. The small, pale yellowish flowers (0.3-0.4 cm diameter) are functionally unisexual and organized in dense inflorescences (ca. 45 flowers). P. spruceanum presented annual flowering between September and November. Staminate flowers supplied a high percentage of viable pollen (90.6%) and relatively abundant nectar (x = 4.5 μL). Pistillate flowers produced only nectar to flower visitors (x = 4.0 μL). The effective pollinators were Apis mellifera and Trigona sp. (Hymenoptera, Apidae). Pollen tubes of cross-pollinated flowers were observed entering the ovaries 48 h after pollination. The fruiting season is from October to March, with a peak in November, coinciding with the rainfall peak. Ecological implications of these findings, and alternative arguments to explain the high genetic diversity at regional landscape are discussed.

USE OF MUELLER-HINTON BROTH AND AGAR IN THE GERM TUBE TEST

Candida albicans is often isolated from clinical samples, thus its presumptive differentiation from other species of the same genus can be based on its ability to form the germ tube in human serum. Nevertheless, there are two other species that share this characteristic: C. dubliniensis and C. africana. The aim of this study was to compare four different substrates to perform the germ tube (GT) test. The Candida spp. isolates were identified using a manual system (135 C. albicans, 24 C. tropicalis and one C. dubliniensis). The germ tube test was performed with fresh, previously frozen serum and Mueller-Hinton (MH) broth and agar. GT was observed in 96% (130/136) of the isolates through the fresh serum technique, 94% (128/136) through previously frozen serum, 92% (125/136) in MH agar, and 90% (122/136) in MH broth. The sensitivity of each test was higher than 90%, with 100% specificity. Both the MH agar and broth were able to identify the true positives, and false positives were not found. However, some C. albicans isolates were not identified. MH agar and broth may be used in laboratory for the rapid presumptive identification of C. albicans, as an alternative method for germ tube test.

Determination of germ tube, phospholipase, and proteinase production by bloodstream isolates of Candida albicans

Introduction Candida albicans is a commensal and opportunistic agent that causes infection in immunocompromised individuals. Several attributes contribute to the virulence and pathogenicity of this yeast, including the production of germ tubes (GTs) and extracellular hydrolytic enzymes, particularly phospholipase and proteinase. This study aimed to investigate GT production and phospholipase and proteinase activities in bloodstream isolates of C. albicans. Methods One hundred fifty-three C. albicans isolates were obtained from blood samples and analyzed for GT, phospholipase, and proteinase production. The assays were performed in duplicate in egg yolk medium containing bovine serum albumin and human serum. Results Detectable amounts of proteinase were produced by 97% of the isolates, and 78% of the isolates produced phospholipase. GTs were produced by 95% of the isolates. A majority of the isolates exhibited low levels of phospholipase production and high levels of proteinase production. Conclusions Bloodstream isolates of C. albicans produce virulence factors such as GT and hydrolytic enzymes that enable them to cause infection under favorable conditions.

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

Introduction Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. Methods We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. Results The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. Conclusions The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.

Effects of Rosmarinus officinalis essential oil on germ tube formation by Candida albicans isolated from denture wearers

Introduction The aim of this study was to investigate the effects of Rosmarinus officinalis essential oil on germ tube formation by Candida albicans isolated from denture wearers. Methods Ten C. albicans isolates recovered from denture wearers were tested using 10% fetal bovine serum with or without 4% R. officinalis essential oil. Results The essential oil from R. officinalis completely inhibited germ tube formation in the investigated C. albicans isolates. Conclusions The results demonstrate that the essential oil of R. officinalis modulates C. albicans pathogenicity through its primary virulence factor (i.e., germ tube formation was suppressed).

Single-tube nested PCR assay with in-house DNA extraction for Mycobacterium tuberculosis detection in blood and urine

Abstract: INTRODUCTION : Molecular analyses are auxiliary tools for detecting Koch's bacilli in clinical specimens from patients with suspected tuberculosis (TB). However, there are still no efficient diagnostic tests that combine high sensitivity and specificity and yield rapid results in the detection of TB. This study evaluated single-tube nested polymerase chain reaction (STNPCR) as a molecular diagnostic test with low risk of cross contamination for detecting Mycobacterium tuberculosis in clinical samples. METHODS: Mycobacterium tuberculosis deoxyribonucleic acid (DNA) was detected in blood and urine samples by STNPCR followed by agarose gel electrophoresis. In this system, reaction tubes were not opened between the two stages of PCR (simple and nested). RESULTS: STNPCR demonstrated good accuracy in clinical samples with no cross contamination between microtubes. Sensitivity in blood and urine, analyzed in parallel, was 35%-62% for pulmonary and 41%-72% for extrapulmonary TB. The specificity of STNPCR was 100% in most analyses, depending on the type of clinical sample (blood or urine) and clinical form of disease (pulmonary or extrapulmonary). CONCLUSIONS: STNPCR was effective in detecting TB, especially the extrapulmonary form for which sensitivity was higher, and had the advantage of less invasive sample collection from patients for whom a spontaneous sputum sample was unavailable. With low risk of cross contamination, the STNPCR can be used as an adjunct to conventional methods for diagnosing TB.