Cytochemical Observations on Proteins,Alkaline and Acid Phosphatases,Adenosine Triphosphatase,and 5′-Nucleotidase in Chick Liver Cell Cultures Infected with Trichomonas vaginalis*, †

SYNOPSIS. By means of the ninhydrin-Schiff method for proteins a diffuse reaction as well as one localized in granular inclusions can be shown in the cytoplasm of fibroblasts, epithelial cells, and macrophages in trypsin-dispersed chick liver cell cultures. Nuclei and nucleoli also take the specific stain. A progressive loss of cytoplasmic and nuclear staining occurs in the fibroblasts in cultures infected with a relatively pathogenic strain of T. vaginalis. A loss occurs in epithelial cells in advanced stages of degeneration, but in less damaged cells, while the diffuse reaction disappears, the number and staining intensity of the cytoplasmic inclusions remain unchanged or possibly may increase somewhat. The intensity of the diffuse reaction and the number and size of the characteristic inclusions increase in the active, parasite-free, experimental macrophages, but phagocytes with trichomonads closely applied to their external surfaces and those containing the flagellates within their cytoplasm typically retain only a few weak-staining inclusions. Similar distribution of alkaline and acid phosphatases occurs in preparations treated according to Gomori's and Burstone's methods, except that no nuclear staining is obtained with the latter. Activity of both enzymes is localized primarily in inclusions which are dispersed thruout the cytoplasm of fibroblasts and epithelial cells and tend to accumulate along the cell membranes and around the nuclei. In the course of infection with T. vaginalis there is a progressive loss of alkaline phosphatase from both cell types; however, the acid phosphatase activity increases. In the control macrophages both enzymes are localized in mostly rather large, rounded cytoplasmic inclusions. The number of such inclusions increases in the parasite-free experimental macrophages, but only a few weak-staining granules remainin phagocytes with engulfed trichomonads and in those whose external surfaces are in direct contact with the parasites. The loss of the inclusions is less apparent in macrophages containing degenerated flagellates than in the ones with healthy trichomonads, but regardless of the condition of the parasites, the highest enzymatic activity is found around them. ATPase and 5′-nucleotidase are localized in small granules dispersed thruout the cytoplasm of fibroblasts and epithelial cells. The granules tend to accumulate along the periphery of the cells and around the nuclei. A diffuse cytoplasmic reaction is present in preparations processed for 5′-nucleotidase. Nuclei and nucleoli give positive reactions for both enzymes. In the course of infection with trichomonads, activity of the 2 enzymes declines in both culture cell types. Control macrophages have diffuse cytoplasmic reaction for ATPase and 5′-nucleotidase and these enzymes are localized also in rounded cytoplasmic inclusions. Activity of both enzymes increases in the parasite-free experimental phagocytes, but little if any diffuse staining and only a few characteristic inclusions are left in macrophages with engulfed healthy trichomonads and in those whose external surfaces are invested with the flagellates. The ninhydrin-Schiff-positive inclusions found in the macrophages appear to be the same as some of those which have acid phosphatase activity and may well be identical with the glycolipoprotein bodies noted by us previously. On the grounds of their chemical constitution and behavior it seems likely that the inclusions are lysosomes.     

Genetics of flowering time in bread wheat <Emphasis Type="Italic">Triticum aestivum</Emphasis>: complementary interaction between vernalization-insensitive and photoperiod-insensitive mutations imparts very early flowering habit to spring wheat

Time to flowering in the winter growth habit bread wheat is dependent on vernalization (exposure to cold conditions) and exposure to long days (photoperiod). Dominant Vrn-1 (Vrn-A1, Vrn-B1 and Vrn-D1) alleles are associated with vernalization independent spring growth habit. The semidominant Ppd-D1a mutation confers photoperiod-insensitivity or rapid flowering in wheat under short day and long day conditions. The objective of this study was to reveal the nature of interaction between Vrn-1 and Ppd-D1a mutations (active alleles of the respective genes vrn-1 and Ppd-D1b). Twelve Indian spring wheat cultivars and the spring wheat landrace Chinese Spring were characterized for their flowering times by seeding them every month for five years under natural field conditions in New Delhi. Near isogenic Vrn-1 Ppd-D1 and Vrn-1 Ppd-D1a lines constructed in two genetic backgrounds were also phenotyped for flowering time by seeding in two different seasons. The wheat lines of Vrn-A1a Vrn-B1 Vrn-D1 Ppd-D1a, Vrn-A1a Vrn-B1 Ppd-D1a and Vrn-A1a Vrn-D1 Ppd-D1a (or Vrn-1 Ppd-D1a) genotypes flowered several weeks earlier than that of Vrn-A1a Vrn-B1 Vrn-D1 Ppd-D1b, Vrn-A1b Ppd-D1b and Vrn-D1 Ppd-D1b (or Vrn-1 Ppd-D1b) genotypes. The flowering time phenotypes of the isogenic vernalization-insensitive lines confirmed that Ppd-D1a hastened flowering by several weeks. It was concluded that complementary interaction between Vrn-1 and Ppd-D1a active alleles imparted super/very-early flowering habit to spring wheats. The early and late flowering wheat varieties showed differences in flowering time between short day and long day conditions. The flowering time in Vrn-1 Ppd-D1a genotypes was hastened by higher temperatures under long day conditions. The ambient air temperature and photoperiod parameters for flowering in spring wheat were estimated at 25°C and 12 h, respectively.     

The effects of disturbance on forest structure and diversity at different altitudes in Garhwal Himalaya

The effects of disturbance on forest structure and diversity along an altitudinal gradient in the temperate, sub-tropical, and tropical regions of Garhwal Himalaya were assessed. Each region was further categorized into undisturbed (UD), mildly disturbed (MD), and highly disturbed (HD) sites on the basis of magnitude of disturbance in these forests. On UD sites of temperate, sub-tropical and tropical regions, Quercus leucotrichophora, Anogeissus latifolia and Holoptelea integrifolia were the dominant tree species respectively. The highest values of tree density (1028 ind*hm-2) and total basal cover at breast height (31.70 m2 *hm-2) were recorded for UD site of temperate region, whereas maximum species diversity (3.128) and equitability (14.09) values were observed for HD site of tropical region. The structure and composition of the forests were greatly affected by the degree of disturbance.     

Changes in Kit Fox Defecation Patterns During the Reproductive Season: Implications for Noninvasive Surveys

Abstract: Noninvasive survey methods based on analyzing DNA extracted from feces can be useful for carnivores that are difficult to study by other methods. Changes in fecal deposition patterns associated with reproduction in kit foxes (Vulpes macrotis) might affect results of such surveys. We used a trained dog to collect fresh scats on 2-km transects in the home ranges of 11 radiocollared female kit foxes in January, February, and March 2008 and determined sex of the individual that deposited the scats by amplifying the zinc finger protein gene. Female foxes give birth in mid-February to mid-March. We found a similar number of scats each month. In January, the sex ratio of the scats was not different from the expected 1:1. However, in February there were almost 2 male scats for every female scat and in March there were >8 male scats for every female scat. Comparing March to January, there were more male scats on all 11 transects and fewer female scats on 10 of 11 transects. Around the time pups are born, both sexes appear to show changes in fecal deposition patterns that make it easier to find male scats and harder to find female scats. Effects of these changes on survey results will vary depending on the purpose and design of the survey. Surveys to determine distribution and relative abundance would probably not be negatively affected by these changes. However, if surveys to estimate abundance are conducted during the reproductive season, they could result in an underestimate of population size unless the increased heterogeneity in scat detectability is taken into account.     

Development and use of anchored‐SSRs to study DNA polymorphism in bread wheat (Triticum aestivum L.)

In bread wheat, 21 anchored simple sequence repeat (SSR) primer pairs detecting SSR length polymorphism and 42 anchored SSR primers detecting microsatellite‐anchored fragment length polymorphisms (MFLPs) are reported. Eight bread wheat genotypes were used for detecting polymorphism. The number of alleles in SSR analysis ranged from two to six, with a mean of 2.9 alleles per SSR. The number of polymorphic bands in MFLP ranged from two to 40, with a mean of 12.74 polymorphic bands/primer combination, the SSRs with CT/GA motifs giving the highest level of polymorphism (a mean of 18.37 bands). The average value of polymorphic information content (PIC) was 0.473 for SSRs and 0.061 for MFLP.     

Effect of inter-cropping bell-pepper with ginger on plant parasitic nematode populations and crop yields

During 1994 studies were undertaken to improve ginger (Zingiber officinale Roscoe) yield against the nematodes Pratylenchus penetrans, Meloidogyne incognita, Helicotylenchus dihystera and Tylenchorhynchus mashhoodi in Himachal Pradesh (HP) (India) by inter-cropping bell-pepper (Capsicum annum L) in eight different sequences (treatments). Inter-cropping of “one rhizome of ginger x one plant of bell-pepper” gave the highest ginger yield (600 g per rhizome). This treatment was completely free from P. penetrans and M. incognita. All treatments with bell-pepper plants equal to or higher in number to that of ginger rhizomes had higher ginger yields than treatments with ginger alone or with fewer bell-pepper plants. In the former, populations of P. penetrans and M. incognita were lower than in the latter treatments. The yield of ginger varied irrespective of population densities of H. dihystera and T. mashhoodi, indicating that P. penetrans and M. incognita are the major nematode problems of ginger in HP. Bell-pepper was a non-host to P. penetrans and non-preferred host to M. incognita. This helped to improve ginger yields by making the rhizosphere unfavourable for the development and multiplication of the major ginger nematode pests.     

A screening technique to evaluate pigeonpea for resistance to Rotylenchulus reniformis*

Rotylenchulus reniformis is one of the most important nematode pests of pigeonpea. A simple greenhouse technique has been developed to aid evaluation of pigeonpea genotypes for resistance to R. reniformis. In greenhouse pot experiments, eggsacs of R. reniformis in pigeonpea (cv. ICPL 87) roots were counted by eye and with the aid of a stereoscopic microscope at 15, 30 and 45 days after seedling emergence in soils infested with various numbers of vermiform R. reniformis. Seedlings were rated for the number of eggsacs per root system on a one (no eggsacs) to nine (more than 50 eggsacs) scale. Eggsac ratings were more uniform when roots were evaluated at 30 – 45 days than at 15 days and an inoculum of 15 to 30 individuals/cm³ soil also helped reduce variability. Eggsacs were not easily visible without the aid of a stereoscopic microscope. Of the 14 stains tested, exposure of nematode-infected roots to 0.25% trypan blue for three min was effective in staining the eggsacs blue without staining the roots. Using the stain, the assessment of infestation by R. reniformis was equally accurate with or without the aid of a stereoscopic microscope. Exposure of eggsacs to trypan blue enhanced the emergence of juveniles from the eggsacs.     

Components of resistance to the sorghum midge, Contarinia sorghicola

Studies were conducted on components of resistance to sorghum midge on four resistant (DJ 6514, AF 28, TAM 2566 and IS 15107) and two susceptible cultivars (CSH 1 and Swarna). Data were recorded on the numbers of eggs, larvae, emerged adults and grain damage in panicles of different genotypes infested with 60 midge females/panicle under no-choice conditions. The size of floral parts (glume, lemma, palea, lodicule, stigma, style, ovary and anther), rate of grain development and tannin content of grain were measured. The lengths of glume gl and 82, lemma L1 and L2, palea, lodicule, anther, style and stigma were positively associated with susceptibility to sorghum midge. Rate of grain development (between 3rd and 7th day after anthesis) was negatively associated with susceptibility to sorghum. Tannin content of grain was also negatively correlated with midge susceptibility, although there were distinct exceptions (e.g. DJ 6514 is highly resistant bur has a low tannin content).     

Antibiosis component of resistance in sorghum to sorghum midge, Contarinia sorghicola

Sorghum midge, Contarinia sorghicola Coq. (Diptera: Cecidomyiidae) is an important pest of grain sorghum, and host-plant resistance is one of the most effective means of controlling this pest. We studied the antibiosis mechanism of resistance in sorghum to C. sorghicola in a diverse array of midge-resistant and midge-susceptible genotypes. Data were recorded on adult emergence, postembryonic developmental period, number of mature eggs in the ovary, fecundity, larval survival from artificially implanted eggs; and the tannins, soluble sugars, and protein content of 10-day old and mature grains during the 1982-91 rainy and post-rainy seasons. Adult emergence was significantly lower in the midge-resistant genotypes compared with the susceptible controls. Initiation of adult emergence was delayed by 4–8 days on DJ 6514, IS 8571, IS 9807, IS 10712, IS 19474, IS 19512, ICSV 830 and ICSV 197. Postembryonic developmental period was prolonged on DJ 6514, IS 15107, IS 3461, IS 7005, IS 19474, ICSV 831 and ICSV 197. However, the delay in adult emergence or the extended developmental period was not observed during the post-rainy season in some genotypes. These differences in the expression of antibiosis to midge in resistant genotypes over seasons may be attributed to the effect of environmental conditions on the insect development and chemical composition of sorghum grain. Amounts of tannins and proteins were generally greater in the midge-resistant lines compared with the susceptible ones (except tannins in DJ 6514) while the soluble sugars were low in the midge-resistant lines (except TAM 2566). These differences in chemical composition of the grain between genotypes and variations over seasons have been discussed in relation to the expression of antibiosis mechanism of resistance to the sorghum midge. Antibiosis to sorghum midge was also evident in terms of smaller size of larvae, lower number of eggs in the ovary, reduced fecundity, and larval survival. Midge-resistant lines have diverse effects on the biology of this insect. Antibiosis along with other components of resistance can be used to develop cultivars with stable resistance to C. sorghicola.