Inhibition of Water Splitting Increases the Susceptibility of Photosystem II to Photoinhibition

Photosystem II (PSII)-enriched membrane particles were isolated from peas (Pisum sativum L.) and treated in several different ways to inhibit the water oxidation reactions, but not reaction center function itself, as judged by the light-induced rate of reduction of 2,6-dichlorophenol indophenol with and without the artificial electron donor, diphenyl carbazide. It was shown that such treatments increased the susceptibility of the PSII-enriched membranes to photoinhibition. This trend was further observed if 2,6-dichlorophenol indophenol was present during the illumination with photoinhibitory light. On the other hand, protection against the enhanced photoinhibition was found when the water-splitting activity was reconstituted or when the artificial electron donor diphenyl carbazide was present during the preillumination. The results indicate that irreversible photodamage occurred within the PSII reaction center as a consequence of illumination with strong light and that the rate of this damage was enhanced under conditions that are expected to give rise to a photoaccumulation of oxidizing species such as P680⁺ on the donor side of PSII. This mechanism of photoinhibitory damage occurred under both aerobic and anaerobic conditions.     

Effect of Cold Treatments on the Binding Stability of Photosystem II Extrinsic Proteins and an Associated Increase in Susceptibility to Photoinhibition

When pea plants (Pisum sativum L. cv Feltham First) are subjected to freezing conditions (−18°C) followed by a thaw to 18°C, there is a significant inhibition of water-splitting capacity judged by the rate of light-induced reduction of 2,6-dichlorophenol indophenol using isolated thylakoid membrane fragments enriched in photosystem II (PSII). The freeze-thaw-induced inhibition of water-splitting activity has been correlated with the loss of the 17- and 23-kilodalton extrinsic protein of PSII and with a weakening of the binding of the 33-kilodalton protein. There was no apparent loss of bound manganese. Addition of 10 millimolar CaCl₂, however, allowed a full recovery of the water-splitting activity of these modified PSII-enriched particles. The freeze-thaw-induced changes in the organization and functional capacity of PSII was found to increase its susceptibility to photoinhibition in agreement with the concepts presented in the accompanying paper, that oxidative damage can occur within the PSII reaction center as a consequence of extending the lifetime of P680⁺.     

The dilemma of negative analysis of variance estimators of intraclass correlation

Summary At least two common practices exist when a negative variance component estimate is obtained, either setting it to zero or not reporting the estimate. The consequences of these practices are investigated in the context of the intraclass correlation estimation in terms of bias, variance and mean squared error (MSE). For the one-way analysis of variance random effects model and its extension to the common correlation model, we compare five estimators: analysis of variance (ANOVA), concentrated ANOVA, truncated ANOVA and two maximum likelihood-like (ML) estimators. For the balanced case, the exact bias and MSE are calculated via numerical integration of the exact sample distributions, while a Monte Carlo simulation study is conducted for the unbalanced case. The results indicate that the ANOVA estimator performs well except for designs with family size n = 2. The two ML estimators are generally poor, and the concentrated and truncated ANOVA estimators have some advantages over the ANOVA in terms of MSE. However, the large biases may make the concentrated and truncated ANOVA estimators objectionable when intraclass correlation () is small. Bias should be a concern when a pooled estimate is obtained from the literature since <0.05 in many genetic studies.     

Maturation and fertilization of porcine oocytes in vitro

Four experiments were conducted to study 1) factors affecting porcine oocyte maturation in culture medium and 2) a new method for oocyte maturation outside the porcine body. In Experiment 1, five groups of oocytes were cultured in m-TCM199 or m-KRB medium for 24 to 28, 32 to 36 or 40 to 42 hours and then were fertilized in vitro. The cleavage rate (two to four-cell stage) of oocytes cultured for 32 to 36 hours was significantly higher than those of the other oocytes. The results indicate that a suitable culture period for the in vitro maturation of porcine oocytes is 32 to 36 hours. In Experiment 2, four groups of oocytes were cultured in m-KRB or m-KRB supplemented with PFF, PMSG or FSH for in vitro maturation, and the cleavage rates of oocytes were 7.94, 22.56, 30.23 and 23.26%, respectively, after in vitro fertilization. The results show that porcine follicular fluid (PFF) and gonadotrophins added to the culture medium promote porcine oocyte maturation in vitro. In Experiment 3, oocytes were cultured in m-KRB or m-TCM199, supplemented with both gonadotrophin and pocine folliclar fluid for maturation in vitro. After fertilization in vitro, the cleavage rates of oocytes were 26.32 and 27.93% for the two media. The results indicate that the difference between m-KRB and m-TCM199 was insignificant when the media were used to culture porcine oocytes. But there was a significant difference when PFF and gonadotrophins were added to the basic media. In Experiment 4, porcine oocytes were transferred into the reproductive tracts of other animals for maturation. After 34 to 36 hours, the oocytes were collected and fertilized in vitro. The cleavage rates of oocytes were 10.42, 28.45, 3.33 and 36.36%, respectively, for the oocytes matured in mouse uterine horns, rat uterine horns, rat oviducts or rabbit oviducts. The results show that porcine oocytes can be matured in the reproductive tracts of other animals.     

16S rRNA-based probes and polymerase chain reaction method to detect Listeria monocytogenes cells added to foods.

A rapid polymerase chain reaction (PCR) method was developed for detection of Listeria monocytogenes in foods. This method used a pair of primers based on a unique region in the 16S rRNA sequence of L. monocytogenes, which were previously reported by us to yield a specific nucleic acid probe. Our method included use of a shorter denaturing time, a shorter annealing time, a rapid transition, and an increase in the number of cycles, resulting in good sensitivity. Just 3 h for PCR plus 1 h for electrophoresis was required. Additional time for DNA isolation and DNA hybridization was not needed. This method detected as few as 2 to 20 CFU of L. monocytogenes in pure cultures and as few as 4 to 40 CFU of L. monocytogenes in inoculated (10(8) CFU), diluted food samples. Seven of eight foods, including four poultry products, gave positive results. Only one food sample, soft cheese, gave interference. An internal probe hybridization test was used to confirm that the PCR products were from L. monocytogenes. A specificity test indicated that this PCR method was positive for all 13 strains of L. monocytogenes tested but negative for the other 6 species of Listeria, including 6 strains of L. innocua, and negative for 17 other gram-positive and gram-negative bacteria tested.     

Molecular cloning and mapping of phenol degradation genes from Bacillus stearothermophilus FDTP-3 and their expression in Escherichia coli.

Two genes of the meta pathway of phenol degradation were cloned from a phenol-utilizing strain of Bacillus stearothermophilus and were mapped by subcloning and by use of a Tn5 insertion mutation. They code for phenol hydroxylase and catechol 2,3-dioxygenase, respectively. The gene encoding catechol 2,3-dioxygenase, which is more thermostable than catechol 2,3-dioxygenase encoded by the other gene, shares rather limited homology with that from Pseudomonas putida.     

Enantioselective hydrolysis of hydrophobic amino acid derivatives by lipases

Microbial lipases preferentially cleave the L-isomers of N-benzyloxycarbonyl (Z)-hydrophobic D,L-amino acid methyl esters which is the same as that of subtilisin. This implies that lipases and proteases may share the same ancestor in the evolutionary point and lipases can be practically applied to resolve racemic hydrophobic amino acid derivatives.     

An mRNA that specifically accumulates in maize roots delineates a novel subset of developing cortical cells

A near full-length cDNA clone (pZRP3) corresponding to an mRNA that accumulates specifically in roots of maize was isolated. The ZRP3 mRNA is ca. 600 nucleotides in length. The amino acid sequence of the predicted polypeptide is rich in leucine (16%), proline (11%), and cysteine (8.5%). The zrp3 gene appears to be expressed exclusively in roots, whereas other ZRP3-related genes are expressed in additional organs of the maize plant. In situ hybridization shows that ZRP3 mRNA accumulation is largely confined to the cells of the cortical ground meristem. Furthermore, accumulation of this mRNA occurs within a distinct subset of cortical cells, the inner three to four cell layers.Journal paper number J-14572 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa Project Number 2997.     

鼠伤寒沙门氏菌维生素B2生物合成的调节

7 independent rib genes fusions with MudJ (lacZ, Kanr) were isolated by transposon MudJ mutagenesis in Salmonella typhimurium. 5 of them are blue on the X-gal plate, and the beta-galactosidase activity of the cells grown in E medium containing various concentration of riboflavin were assayed. The results showed that the expression of rib gene are not repressed by riboflavin. It appears to be synthesized constitutively in Salmonella typhimurium.