Physiological and genomic variations in rice cells recovered from direct immersion and storage in liquid nitrogen

The use of cryoprotectants and slow cooling rates are routine procedures for the cryopreservation of plant cell lines. However, our results with rice (Oryza sativa L,, ev. Taipei 309) show that calli can be cryopreserved by direct immersion and stored in liquid nitrogen without any cryoprotection, the efficiency of recovery using this method, as well as a conventional method was generally increased with a previous abscisic acid (ABA) treatment. Following cryopreservation, calli demonstrated some differences with respect to unfrozen calli of the same lines, Thus, resistance to freezing stress (- 20 degrees C for 2 h) increased significantly in all lines tested, irrespective of their pre-incubation with ABA, Calli that had been directly stored in liquid nitrogen also demonstrated a higher competence for genetic transformation than their unfrozen counterparts, as indicated by the transient gene expression levels obtained after particle bombardment, These differences might lead to further biotechnological applications, A genetic analysis of amplified DNA polymorphisms was performed with three independent lines that had been subjected to four combinations of ABA treatment and direct immersion in liquid nitrogen, At the loci screened with the randomly amplified polymorphic DNA (RAPD) markers tested, the genetic variations among lines and among calli of the same line appear to bd more related to tissue-culture-induced somaclonal variation than to cryoselection.

Osmocondicionamento de sementes de salsa (Petroselinum sativum Hoffm.) em diferentes potenciais hídricos

A germinação de sementes de salsa (Petroselinum sativum Hoffm.) é lenta e desuniforme, sendo a contagem do teste padrão de germinação recomendada aos 10 e 28 dias e a emergência em campo é relativamente longa, justificando o uso de técnicas que acelerem e uniformizem o processo germinativo: um método promissor é o osmocondicionamento. No presente trabalho, objetivou-se avaliar os efeitos dos diferentes potenciais de osmocondicionamento, com PEG 6000, na germinação de sementes de salsa e identificar o potencial osmótico ótimo para aumentar a germinação destas sementes. Sementes das cultivares Lisa e Portuguesa de dois lotes, adquiridas no comércio local foram osmocondicionadas pelo método de imersão direta das mesmas em soluções aquosas com potencial hídrico: 0,0, -0,5, -1,0 e -1,5 MPa, obtidos com a adição de PEG 6.000, sob aeração constante a 25ºC por 144 horas, como testemunha utilizaram-se sementes não osmocondicionadas. As variáveis analisadas foram: porcentagem de germinação (10 e 28 dias), velocidade de germinação e índice de velocidade de germinação. O osmocondicionamento aumenta a porcentagem e a velocidade de germinação de sementes de salsa, sendo que, o potencial osmótico indicado situa-se entre -1,0 e -1,5 MPa.

Analysis of Steroids using Solid Phase Microextraction-Gas Chromatography-Mass Spectrometry-Mass Spectrometry (SPME-GC-MS-MS)

Direct immersion SPME-GC-MS-MS was used for the analysis of steroids in water at part-per-trillion(ppt) and lower concentrations. The method was validated and extended to real sample analysis. The method were linear from 0.01 to 5 ng/ml with precision less than 10% relative standard deviation for a steroid mixture at 1 ng/ml. Limit of quantitation and limit of detection was found to be 200- 1200 pg/L and 30-200 pg/L respectively and recoveries ranged from 88-103 %. To understand the extraction efficiency of the fiber, a depletion study was performed. The fiber/ sample partition coefficients for the steroids were determined to be 1.0 x 104 to 1.5 x 104 . The extraction was performed without derivatization or the use of an internal standard. SPMEGC-MS-MS effectively demonstrated ultra-trace level detection of steroids in water.

Hardness and compressive strength of indirect composite resins: effects of immersion in distilled water

The aim of this study was to evaluate the effect of ageing in distilled water on the hardness and compressive strength of a direct composite resin Z100, a feldspatic porcelain (Noritake) and three indirect composites (Artglass, Solidex and Targis). For the Vickers hardness tests, five disk-shaped specimens (2 x 4 mm) of each material were prepared according to the manufacturers' instructions. The hardness tests were conducted using a Vickers diamond indentor. Compressive strength measurements were recorded on cylindrical specimens with a diameter of 6 mm and a length of 12 mm. The compression tests were carried out with a constant cross-head speed of 0.5 mm min(-1) on a mechanical test machine. For each material, 10 specimens were tested after 7 days of dry storage at 37 +/- 1 degreesC and 10 specimens were tested after water storage at 37 +/- 1 degreesC for 180 days. Noritake porcelain specimens showed higher hardness values than the composites. Among the composite materials, Z100 promoted the highest VHN values, regardless of the ageing periods. The results showed that Solidex and Z100 had the highest compressive strength values. Ageing in water reduced the hardness for all composites, but had no long-term effect on the compressive strength.