Meropenem as an Alternative Antibiotic Agent for Suppression of Agrobacterium in Genetic Transformation of Orchid

A case of Meropenem as a novel antibacterial agent to suppress and eliminate Agrobacterium tumefaciens in the Agrobacterium-mediated transformation of orchid protocorm-like bodies （PLBs） has been reported in this article. The in vitro activities of meropenem and four comparator antibacterial agents against three Agrobacterium tumefaciens strains, LBA4404, EHA101, and GV3101, were assessed. In addition, the effect of meropenem on the growth of Dendrobium phalaenopsis PLBs was determined. Compared with other commonly used antibiotics （including ampicillin, carbenicillin, cefotaxime, and cefoperazone）, meropenem showed the highest activity in suppressing all tested A. tumefaciens strains （minimum inhibitory concentration [MIC] 〈 0.5 mg L^-1, which is equal to minimum bactericidal concentration [MBC]）. Meropenem, at all tested concentrations, except for 10 mg L^-1 concentration, had little negative effect on the growth of orchid tissues. The A. tumefaciens strain EHA101 in genetic transformation with vector plG121Hm in infected PLBs of the orchid was visually undetectable after a two-month subculture in 1/2 MS medium with 50 mg L^-1 meropenem and 25 mg L^-1 hygromacin. The expression and incorporation of the transgenes were confirmed by GUS histochemical assay and PCR analysis. Meropenem may be an alternative antibiotic for the effective suppression of A. tumefaciens in genetic transformation.     

Morphometry on lancet flukes found in Japanese sika deer (Cervus nippon centralis) captured in Iwate Prefecture, Japan

Thirty-six flukes were collected from the livers of wild deer (Cervus nippon centralis) captured in Iwate Prefecture, Japan, and were served for morphometry. The length and/or the width of the body, suckers, testes, ovary, vitelline glands, cirrus pouch and eggs in the uterus of the flukes were measured. The distance between anterior end of the body and position of the maximal body-width or upper end of the testes were also determined. A remarked morphological characteristic was that the right and left testes did not lie tandem but lined bilaterally. Also the position of the maximal body-width did not always locate in the posterior part of the body of the fluke. The property was in accordance with those for Dicrocoelium chinensis.     

Rediscovery of Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) MK.) an ancient crop of the Indian subcontinent

Indian dwarf wheat (Triticum aestivum L. ssp. sphaerococcum (Perc.) Mac Key, synonym: T. sphaerococcum Perc.) is endemic to southern Pakistan and northwestern India. It was one of the main winter crops grown by ancient Indian cultures. However, it disappeared from the record during the early twentieth century, especially after the Green Revolution brought modern wheat varieties into India and Pakistan. Whether or not Indian dwarf wheat is presently cultivated has been unclear. Here we report on the rediscovery of the cultivation of this wheat in northern Karnataka and southern Maharashtra in India. Molecular genetic analysis of the chloroplast DNA of the two specimens collected at location 3 revealed that both samples have a unique haplotype that is specific to Indian dwarf wheat. We found this wheat at three locations in 2010, but at only one of the three locations in 2011. Therefore, the future survival of this subspecies is uncertain. Further ethnobotanical research is urgently needed to conserve this unique genetic resource for the future.     

Production of triploid plants of Japanese pear (Pyrus pyrifolia Nakai) by anther culture

Triploid plants were grown from the diploid pear scion cultivar (Pyrus pyrifolia Nakai) by anther culture. Androgenic callogenesis occurred at about 10 days after anther excision, and most calli formed within four weeks and stopped growing at less than 1–3 mm in diameter. Androgenic embryogenesis occurred within 8–12 weeks after anther culture. The formed embryos were generally white, globular or heart-shaped. To increase callogenesis and embryogenesis, the effects of cold pre-treatment, light and activated charcoal (AC) on anthers collected from buds were investigated. Cold pre-treatment, dark and lack of AC increased callogenesis more than no cold pre-treatment, light and adding AC. A decisive condition was not found for embryogenesis. The embryogenesis frequency was extremely low and only six embryos were grown. Two embryos expanded greenish cotyledons and developed foliage, and they proliferated when transferred monthly to fresh proliferation medium. The proliferated shoots were rooted and acclimated. Flow cytometric analysis and investigation of the number of chromosomes of regenerants showed that all cells were triploid.     

Dietary carbohydrate and growth,body composition and heat increment in rainbow trout (Salmo gairdneri)

A study was conducted to determine growth, body composition and heat increment (HI) of rainbow trout reared on isonitrogenous high digestible carbohydrate (HC) and high fat (HL) diets at 15°C. Trout reared on the HC diet had a significantly lower final body weight and carcass lipid content and a significantly higher feed:gain ratio and carcass protein content than trout reared on the HL diet after 12 weeks on the test diets. The lower carcass lipid composition indicates that trout do not readily convert dietary carbohydrates into fat. HI of trout reared on the HC diet was significantly higher than that of fish reared on the HL diet. Considering the poor utilization of dietary carbohydrates as an energy source and the apparently low conversion of dietary glucose into fat, the cause of the higher HI associated with the HC diet is not completely understood. However, it is possible the elimination of excess dietary glucose is an energy dependent process resulting in an increased heat production which would partly account for the increased HI. Furthermore, if the HC diet is a low net energy diet, then the metabolism and utilization of dietary and tissue proteins (amino acids) for energy by the trout may also be partly responsible for the increased HI of the trout reared on the HC diet.     

Influence of low and high temperatures on the initiation and the development of a bulb primordium in isolated tulip embryos

The influence of 4°C and 24°C on the germination and the subsequent development of isolated tulip embryos was studied, using embryo culture techniques. A low temperature not only influenced the breaking of the embryo dormancy, but also the initiation of a bulb primordium. High-temperature treatment after chilling accelerated the development of an initiated bulb primordium.     

Suspected spontaneous hyperadrenocorticism in a young experimental beagle dog

A 6-month-old female beagle dog, assigned to the low-dose group in a toxicity study, was evaluated for compound toxicity, and spontaneous hyperadrenocorticism was suspected. The animal had an externally apparent distended abdomen on clinical examination upon arrival. Pre-dose clinical pathology showed slightly higher erythroid parameters and stress leukogram on hematology; plasma biochemistry showed higher total protein, gamma-glutamyl transferase, total cholesterol, and triglyceride levels than the reference data. On necropsy, a prominent increase in adipose tissues of the subcutis and abdomen and increased weight of the adrenal gland and liver were observed. Histopathology revealed diffuse hyperplasia of adrenocortical cells in the zona fasciculata and reticularis, cortical atrophy of the thymus, and abundant glycogen accumulation in the hepatocytes. These findings were incidental and not test-substance-related. Electron microscopy of the adrenocortical cells in the zona fasciculata revealed decreased typical translucent lipid droplets, increased electron-dense lipid droplets, and abundant smooth endoplasmic reticulum and lysosomes. Additionally, increased numbers of various sizes and forms of mitochondria with tubular, vesicular, or lamellar cristae compared to that of normal animals were observed. These ultrastructural characteristics of the adrenocortical cells suggested hyperfunction. The pre-dose plasma cortisol levels were slightly higher than those of other females assigned to the toxicity study, while plasma adrenocorticotropic hormone levels were within the normal range. These findings indicate that hyperadrenocorticism is a possible cause of the systemic changes in this case.     

Yield-enhancing and tuber-downsizing effects of transplantation cultivation method of case-held tuber seedlings in the sweet potato cultivar Beniharuka

We developed transplantation cultivation method of case-held tuber seedlings (CTS), which was derived from direct planting method of seed tubers, and applied this method to the sweet potato cultivar Beniharuka. A plastic case made of polypropylene was designed for cultivation of CTS. Seed tubers of cultivar Beniharuka in the range of 30–80 g were cut in half. The half-cut tubers were placed inside the plastic cases, and the cases were filled with a commercial soil mix. The case-held tubers were incubated under natural sunlight in a glass house. After 3–4 wk, the CTS were transplanted into a field. Mother tuber (seed tuber) enlargement was suppressed by the plastic confinement of the cases, and daughter tubers were formed above the case as vine-root-originated tubers. In the field experiments in 2012 and 2013, daughter tuber yields were increased 19% and 21% by case-held tuber seedling transplanting (CTST) over conventional vine-planting (VP), the number of daughter tubers per plant in CTST were 36 and 68% higher than in VP, and the mother tuber yields were limited to 2.1 and 4.3% of the total fresh yield of mother and daughter tubers, respectively in 2012 and 2013. Application of CTST method to cultivar Beniharuka enhanced tuber yield, increased the number of daughter tubers per plant, downsized daughter tubers compared to VP, and mother tuber enlargement was suppressed by case-holding. The CTST method is expected to produce more and smaller good in shape tubers of cultivar Beniharuka compared to VP.