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Objective: The effects of semi-solid enteral formula were investigated in tube feeding patients with aspiration pneumonia and/or diarrhea caused by liquid enteral formula. Methods: In 25 cases of aspiration pneumonia and 10 cases of diarrhea (5 cases had both aspiration pneumonia and diarrhea) caused by liquid enteral formula, the rate of improvement by changing the liquid enteral formula to semi-solid enteral formula was studied. The semi-solid enteral formula (PG Soft® EJ) was infused via the nasogastric tube (16Fr) or percutaneous endoscopic gastrostomy (PEG) tube (20Fr). Results: The semi-solid enteral formula was effective in 72% of aspiration pneumonia cases and in 80% of diarrhea cases. Constipation was observed in one case but was controlled with magnesium oxide. Conclusion: In cases of aspiration pneumonia and/or diarrhea, changing liquid enteral formula to semi-solid enteral formula frequently shows improvement.

Estracyt○R (estramustine phosphate) is a medical drug for prostate cancer with cytotoxic activity causing disruption of microtubule organization and indirect androgen production suppressing activity by its metabolite, estradiol. Based on the data obtained from the Estracyt○R Special Drug Use Investigation which surveyed the clinical efficacy and safety of Estracyt○R in patients with prostate cancer whose relapse of prostate cancer after combined androgen blockade (CAB) therapy was confirmed, we evaluated the progression-free survival, prognostic factor, decrease in prostate specific antigen (PSA) level and safety. This surveillance was conducted at 147 institutions nationwide between October, 2010 and September, 2013 and clinical efficacy was evaluated in 239 cases and safety in 329 cases. The median duration of progression-free survival, PSA progression-free survival and PSA response were 169 days (95%CI, 142-190), 197 days (95%CI, 169-267) and 385 days, respectively. The decrease in PSA level was observed in 125 cases (52.3%). Rate of PSA decline ＞50 and ＞25% were 18.4 and 43.1, respectively, and rate of PSA best response (PSA decline ＞ 50%) was 32.6%. Multivariate analysis demonstrated that long duration of prior CAB therapy, Estracyt○R - pretreatment PSA value and bone metastasis influenced progression-free survival significantly. Adverse events were observed in 127 cases (38.6%). The major adverse events were anorexia which was observed in 35 cases (10.9%), gastrointestinal disorders observed in 32 cases (9.7%), abnormal laboratory test values observed in 31 cases (9.4%) and gynecomastia observed in 16 cases (4.9%). These results suggest the clinical efficacy and safety of Estracyt○R for chemotherapy-naïve castration-resistant prostate cancer (CRPC), and Estracyt○R is regarded as one of the treatment options for patients with CRPC, especially for patients who had long duration of prior CAB therapy.

We investigated the UVB-sensitivity in 12 rice strains belonging to two cultivated species (O. sativa and O. glaberrima) and three wild species (O. barthii, O. meridionalis and O. rufipogon) of rice possessing the AA genome, while focusing on the CPD photolyase activity and the genotypes of CPD photolyase. Although the UVB sensitivity, CPD photolyase activity, and CPD photolyase genotype varied widely among these rice species, the sensitivity to UVB radiation depended on the activity of the CPD photolyase, regardless of grass shape, habitat, or species. The rice strains examined here clearly divided into three groups based on the CPD photolyase activity, and the activity of the strains greatly depended on amino acid residues at positions 126 and 296, with the exception of the W1299 strain (O. meridionalis). The amino acid residues 126 and 296 of CPD photolyase in Sasanishiki strain (O. sativa), which showed higher enzymatic activity and more resistance to UVB, were glutamine (Gln) and Gln, respectively. An amino acid change at position 126 from Gln to arginine ("Nori"-type) in the photolyase led to a reduction of enzymatic activity. Additionally, an amino acid change at position 296 from Gln to histidine led to a further reduction in activity. The activity of the W1299 strain, which possesses a "Nori"-type CPD photolyase, was the highest among the strains examined here, and was similar to that of the Sasanishiki. The CPD photolyase of the W1299 contains ten amino acid substitutions, compared to Sasanishiki. The alterations in amino acid residues in the W1299 CPD photolyase compensated for the reduction in activity caused by the amino acid substitutions at positions 126. Knowledge of the activity of different CPD photolyase genotypes will be useful in developing improved rice cultivars.

Rice cultivars vary widely in their sensitivity to ultraviolet B (UVB) and this has been correlated with cyclobutane pyrimidine dimer (CPD) photolyase mutations that alter the structure/function of this photorepair enzyme. Here, we tested whether CPD photolyase function determines the UVB sensitivity of rice (Oryza sativa) by generating transgenic rice plants bearing the CPD photolyase gene of the UV-resistant rice cultivar Sasanishiki in the sense orientation (S-B and S-C lines) or the antisense orientation (AS-D line). The S-B and S-C plants had 5.1- and 45.7-fold higher CPD photolyase activities than the wild-type, respectively, were significantly more resistant to UVB-induced growth damage, and maintained significantly lower CPD levels in their leaves during growth under elevated UVB radiation. Conversely, the AS-D plant had little photolyase activity, was severely damaged by elevated UVB radiation, and maintained higher CPD levels in its leaves during growth under UVB radiation. Notably, the S-C plant was not more resistant to UVB-induced growth inhibition than the S-B plant, even though it had much higher CPD photolyase activity. These results strongly indicate that UVB-induced CPDs are one of principal causes of UVB-induced growth inhibition in rice plants grown under supplementary UVB radiation, and that increasing CPD photolyase activity can significantly alleviate UVB-caused growth inhibition in rice. However, further protection from UVB-induced damage may require the genetic enhancement of other systems as well.

Class I and class II CPD photolyases are enzymes which repair pyrimidine dimers using visible light. A detailed characterization of class I CPD photolyases has been carried out, but little is known about the class II enzymes. Photolyases from rice are suitable for functional analyses because systematic breeding for long periods in Asian countries has led to the selection of naturally occurring mutations in the CPD photolyase gene. We report the biochemical characterization of rice mutant CPD photolyases purified as GST-form from Escherichia coli. We identified three amino acid changes, Gln126Arg, Gly255Ser, and Gln296His, among which Gln but not His at 296 is important for complementing phr-defective E. coli, binding UV-damage in E. coli, and binding thymine dimers in vitro. The photolyase with Gln at 296 has an apoenzyme:FAD ratio of 1 : 0.5 and that with His at 296 has an apoenzyme:FAD ratio of 1 : 0.12-0.25, showing a role for Gln at 296 in the binding of FAD not in the binding of thymine dimer. Concerning Gln or Arg at 126, the biochemical activity of the photolyases purified from E. coli and complementing activity for phr-defective E. coli are similarly proficient. However, the sensitivity to UV of cultivars differs depending on whether Gln or Arg is at 126. The role of Gln and Arg at 126 for photoreactivation in rice is discussed.

*Summary Sensitivity to ultraviolet-B (UVB) radiation (280–320 nm) varies widely among rice cultivars. We previously indicated that UV-resistant rice cultivars are better able to repair cyclobutane pyrimidine dimers (CPDs) through photorepair than are UV-sensitive cultivars. In this paper, we report that UVB sensitivity in rice, in part, is the result of defective CPD photolyase alleles. Surjamkhi (indica) exhibited greater sensitivity to UVB radiation and was more deficient in CPD photorepair ability compared with UV-resistant Sasanishiki (japonica). The deficiency in CPD photorepair in Surjamkhi resulted from changes in two nucleotides at positions 377 and 888 in the photolyase gene, causing alterations of two deduced amino acids at positions 126 and 296 in the photolyase enzyme. A linkage analysis in populations derived from Surjamkhi and Sasanishiki showed that UVB sensitivity is a quantitative inherited trait and that the CPD photolyase locus is tightly linked with a quantitative trait locus that explains a major portion of the genetic variation for this trait. These results suggest that spontaneously occurring mutations in the CPD photolyase gene cause different degrees of sensitivity to UVB in rice, and that the resistance of rice to UVB radiation could be increased by increasing the photolyase function through conventional breeding or bioengineering.

Variation in growth, grain size and grain storage protein content of rice (Oryza sativa L.) in response to elevated UV-B radiation under sunlight was examined in a cool rice-growing region of Miyagi Prefecture, Japan, in 1999, 2001 and 2002. Tiller number, dry mass, panicle number, grain yield and grain size significantly decreased under elevated UV-B radiation in 2001 and 2002. The effects of elevated UV-B radiation on the reduction of each growth parameter were greatly enhanced by daily lower temperature during the ripening stage in those two years. On the contrary, total grain nitrogen content and grain storage protein content significantly increased under elevated UV-B radiation in 2001 and 2002. Among grain storage proteins, glutelin content significantly increased but albumin-globulin and prolamin contents did not. It was thus evident that not only grain size but also grain storage protein of rice was markedly influenced due to elevated UV-B radiation.

Growth of a near-isogenic line (NIL) for the purple leaf gene Pl of rice with a genetic background of Taichung 65 (T-65) rice was significantly retarded by supplementary ultraviolet-B radiation (UV-B), despite the fact that the amounts of UV-absorbing compounds and anthocyanins in NIL were significantly higher than those in T-65. In order to understand the role of flavonoids in UV-B induced damage protection in T-65 and the NIL, both the (1) relationships between changes in the steady state of cyclobutane pyrimidine dimer (CPD) levels and changes in accumulation of anthocyanins and UV-absorbing compounds in leaves with leaf age, and (2) the susceptibility to CPD induction by UV-B radiation and the ability to photorepair CPD were examined. Although supplementary UV-B elevated the steady state of CPD levels in leaves in both strains, the level in the leaf of the NIL was higher than that in T-65 at any time. The susceptibility to CPD induction by short-term (challenge) UV-B exposure was lower in the NIL than in T-65. On the other hand, the CPD photorepair was also lower in the leaves of the NIL than in those of T-65. The decrease in CPD-photorepair in the NIL was due to a lowering of the leaf-penetrating blue/UV-A radiation, which is effective for photoreactivation by photolyase, by anthocyanins. Thus, accumulation of anthocyanins and UV-absorbing compounds did not effectively function as screening against damage caused by elevated UV-B radiation in the NIL, and the retardation of growth in the NIL resulted from its lower ability to photorepair CPD by higher amounts of anthocyanins.

The detection of quantitative trait loci (QTLs) associated with UV-B resistance in rice should allow their practical application in breeding for such a complex trait, and may lead to the identification of gene characteristics and functions. Considerable variation in UV-B resistance exists within cultivated rice (Oryza sativa L.), but its detailed genetic control mechanism has not been well elucidated. We detected putative QTLs associated with the resistance to enhanced UV-B radiation in rice, using 98 BC(1)F(5) (backcross inbred lines; BILs) derived from a cross between Nipponbare (a resistant japonica rice variety) and Kasalath (a sensitive indica rice variety). We used 245 RFLP markers to construct a framework linkage map. BILs and both parents were grown under visible light with or without supplemental UV-B radiation in a growth chamber. In order to evaluate UV-B resistance, we used the relative fresh weight of aerial parts (RFW) and the relative chlorophyll content of leaf blades (RCC). The BIL population exhibited a wide range of variation in RFW and RCC. Using composite interval mapping with a LOD threshold of 2.9, three putative QTLs associated with both RFW and RCC were detected on chromosomes 1, 3 and 10. Nipponbare alleles at the QTLs on chromosome 1 and 10 increased the RFW and RCC, while the Kasalath allele at the QTL on chromosome 3 increased both traits. Furthermore, the existence of both QTLs on chromosomes 1 and 10 for UV-B resistance was confirmed using chromosome segment substitution lines. Plants with Kasalath alleles at the QTL on chromosome 10 were more sensitive to UV-B radiation than plants with them on chromosome 1. These results also provide the information not only for the improvement of UV-B resistance in rice though marker-associated selection, but also for the identification of UV-B resistance mechanisms by using near-isogenic lines.

Ultraviolet radiation induces the formation of two classes of photoproducts in DNA-the cyclobutane pyrimidine dimer (CPD) and the pyrimidine [6-4] pyrimidone photoproduct (6-4 product). Many organisms produce enzymes, termed photolyases, which specifically bind to these lesions and split them via a UV-A/blue light-dependent mechanism, thereby reversing the damage. These photolyases are specific for either CPDs or 6-4 products. Two classes of photolyases (class I and class II) repair CPDs. A gene that encodes a protein with class II CPD photolyase activity in vitro has been cloned from several plants including Arabidopsis thaliana, Cucumis sativus and Chlamydomonas reinhardtii. We report here the isolation of a homolog of this gene from rice (Oryza sativa), which was cloned on the basis of sequence similarity and PCR-based dilution-amplification. The cDNA comprises a very GC-rich (75%) 5; region, while the 3; portion has a GC content of 50%. This gene encodes a protein with CPD photolyase activity when expressed in E. coli. The CPD photolyase gene encodes at least two types of mRNA, formed by alternative splicing of exon 5. One of the mRNAs encodes an ORF for 506 amino acid residues, while the other is predicted to code for 364 amino acid residues. The two RNAs occur in about equal amounts in O. sativa cells.

The effects of supplementary ultraviolet-B (UV-B) radiation on the changes in synthesis and degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and light-harvesting chlorophyll ...

The aim of this study was to investigate the regulatory effect of ultraviolet-B (UV-B) radiation on a number of key stress response genes found in the epidermis and mesophyll of Pisum sativum L., Argenteum mutant. This mutant was chosen for the ease with which the entire epidermis can be removed from the mesophyll tissue. An additional goal was to explore the potential modifying effect of pre-acclimation of plants to UV-B radiation prior to exposure by UV-B during treatment. Results showed that mRNA accumulation was similar during acute short-term UV-B exposure for chalcone synthase (Chs) and short-chain alcohol dehydrogenase (SadA) in both epidermis and mesophyll. In contrast, the mRNA levels differed considerably between tissues for phenylalanine ammonia lyase, chalcone isomerase and lipid transfer protein. After 24 h incubation in visible light after cessation of UV-B exposure, the regulation of mRNA levels also differed between Chs and SadA, the former showing no expression in the epidermis and the latter none in the mesophyll. Acclimation to low UV-B levels before acute exposures resulted in delayed induction of Chs and SadA. Measurements of UV-B-induced cyclobutane pyrimidine dimers (CPDs) showed a greater formation in epidermis than in mesophyll. In addition, acclimation at low UV-B levels resulted in significantly higher basal levels of CPDs than in non-acclimated plants in both mesophyll and epidermis and also in increased damage in concomitant acute exposures. The lack of correlation between the number of CPDs and levels of transcripts for defence genes, indicates that DNA damage does not control transcription of these genes.

A cDNA clone derived from a novel gene (uvi-1) that is inducible by near-UV light was isolated by a differential screening procedure from a cDNA library of the fungus Bipolaris oryzae and characterized further. Sequence analysis of the clone revealed that uvi-1 encodes a protein with a putative molecular mass of 17 kDa; the UVI-1 protein shows significant similarity to a putative protein encoded by a cDNA which is expressed during appressorium formation in the rice blast fungus, Magnaporthe grisea. The corresponding genomic clone was also isolated, and Southern analysis of genomic DNA indicated the presence of a single copy of the uvi-1 gene in B. oryzae. Northern analysis showed that the uvi-1 transcripts are induced by exposure to near-UV light, but not by blue or red light. Furthermore, accumulation of uvi-1 transcripts is observed during differentiation of the appressorium.

We produced near isogenic lines (NILs) for three purple leaf genes, Pl, Plw and Pli of rice with a genetic back-ground of T-65 to characterize each allele. Three NILs each had characteristic tissue-specific anthocyanin pigmentation. T-65 Plw was newly found to make the root purple when the root was exposed to light. To explore the inducible characteristics of the three genes, we examined the responses of the three NILs to UV-B irradiation. Although the accumulation of anthocyanin increased in all NILs with the increase in the intensity of UV-B irradiation, we observed differential responses for the accumulation of anthocyanin in the NILs: T-65 Pli accumulated the largest amounts of anthocyanin under weak and intermediate intensity of UV-B, though this NIL reduced anthocyanin accumulation under a high intensity of UV-B. T-65 Pl or Plw increased anthocyanin accumulation with increasing intensity of UV-B. Anthocyanin accumulation of the Pl NIL was higher than that of the Plw NIL under each intensity of UV-B. In addition, the total biomass of all the irradiated NILs decreased with the increasing dose of UV-B irradiation and the decrease was negatively correlated with the increase in anthocyanin accumulation in the dose range of UV-B irradiation.

An investigation was made of the variations in growth and grain yield in response to increased exposure to UV-B radiation of Japanese lowland rice ( Oryza sativa L.) in a cool rice-growing region. Two cultivars, UV-resistant cv. ‘Sasanishiki’ and UV-sensitive cv. ‘Norin 1’, were examined in a lowland field at Kashimadai (37°28′E, 141°06′E) in Miyagi Prefecture, Japan, for four cropping seasons from 1994 to 1997. The two cultivars were grown in a lowland field with or without supplemental UV-B radiation, which was provided by UV-B-emitting fluorescent lamps, with a 0.1-mm-thick cellulose diacetate film as a filter. In both cultivars, significant decreases in tiller number as the result of supplemental UV-B radiation were observed during the tillering stage in 1994, 1995 and 1997. Furthermore, decreases in grain size from supplemental UV-B radiation were recorded in all seasons. The trend towards small grain size was pronounced in 1996. In that year, the mean daily middle temperatures were lower throughout most of the cropping season and the mean daily hours of sunshine during the tillering stage and between the end of the panicle differentiation stage and the beginning of the ripening stage were shorter. In 1993 when the temperature and the amount of sunshine were both lower, the tiller number, the dry mass of aboveground parts and the panicle number were significantly reduced by supplemental unfiltered UV-B radiation. There was a cultivar difference in the inhibitory effects of supplemental UV-B radiation on growth between the sensitive cultivar Norin 1 and the resistant cultivar Sasanishiki. These results indicate that supplemental UV-B radiation has a positive effect on the growth and grain development of rice, which may be enhanced by unusual climatic conditions such as lower temperature and less sunshine, in cool rice-growing regions.

Norin 1, a progenitor of many economically important Japanese rice strains, is highly sensitive to the damaging effects of UVB radiation (wavelengths 290 to 320 nm). Norin 1 seedlings are deficient in photorepair of cyclobutane pyrimidine dimers. However, the molecular origin of this deficiency was not known and, because rice photolyase genes have not been cloned and sequenced, could not be determined by examining photolyase structural genes or upstream regulatory elements for mutations. We therefore used a photoflash approach, which showed that the deficiency in photorepair in vivo resulted from a functionally altered photolyase. These results were confirmed by studies with extracts, which showed that the Norin 1 photolyase-dimer complex was highly thermolabile relative to the wild-type Sasanishiki photolyase. This deficiency results from a structure/function alteration of photolyase rather than of nonspecific repair, photolytic, or regulatory elements. Thus, the molecular origin of this plant DNA repair deficiency, resulting from a spontaneously occurring mutation to UV radiation sensitivity, is defective photolyase.

We have examined the changes in the steady-state levels of cyclobutyl pyrimidine dimer (CPD) in rice plants (Oryza sativa L.) growing indoors and outdoors with or without supplemental UV-B radiation. The CPD levels remain between 3 and 6 CPD Mb−1 throughout the day in plants grown under sunlight although the levels fluctuate. The CPD levels are slightly elevated with UV-B (1 W m−2) supplementation, but remain constant (between 5 and 8 CPD Mb−1) throughout the day. In plants grown under visible radiation in a growth chamber, the CPD levels remain constant but are lower compared with those in outdoor-grown plants; they are slightly elevated by supplementation with UV-B radiation. Under exposure to UV-B radiation without visible radiation, the CPD levels gradually increase with lengthening of the irradiation time and reach a level of approximately 35 CPD Mb−1 at dusk. However, this level decreases during the night and then increases again the next day. It is thus evident that the CPD levels are strictly maintained within constant limits throughout the day in plants growing indoors and outdoors with or without supplemental UV-B radiation. It is also found that excision repair occurs during the night in plants having higher CPD levels.

Free radicals with a long lifetime were observed in the leaves of two rice cultivars (Oryza sativa L.), Sasanishiki (UVB resistant) and Norin-1 (UVB sensitive), by electron spin resonance spectroscopy. The leaves of both cultivars grown with visible light show very similar ESR spectra composed of radical 1 (R1) and radical 2 (R2), which may be attributable to P700 cation radicals in the reaction center of photosystem I, and tyrosine cation radicals in the reaction center of photosystem II, respectively. The ESR spectrum composed of R1 and R2 radicals in the leaves of Sasanishiki grown under visible light with supplemental UVB was similar to that in the plant grown without supplemental UVB. On the other hand, the amount of R2 radicals in the leaves of Norin-1 grown under visible light with supplemental UVB was significantly smaller than that in the plant grown without supplemental UVB. It is suggested that the loss of R2 radicals in Norin-1 upon UVB irradiation is related to the instability of the plant.

— We examined the effects of a light environment during culture of rice plants (Oryza sativa) on the steady-state cyclobutyl pyrimidine dimer (CPD) level, CPD induction by challenge UVB exposure and the ability to photorepair CPD. The steady-state CPD level in plants grown under visible radiation with supplemental UVB radiation in a growth chamber was several times higher than in plants grown without supplemental UVB radiation, whereas in outdoor-grown plants, it was not enhanced by supplemental UVB radiation. The susceptibility to CPD induction by challenge UVB exposure was highest in dark-grown plants and decreased with increasing irradiance of visible radiation at low and high levels and outdoors. Chronic UVB radiation reduced the susceptibility to UV-induced CPD in plants grown both indoors and outdoors. There was a significant negative correlation between CPD levels induced by challenge UVB exposure and the content of UV-absorbing compounds. The UV-induced CPD could be reduced by subsequent blue radiation in all samples except in dark-grown seedlings. The higher the irradiance of visible radiation in the culture, the greater the ability to photorepair CPD. Chronic UVB radiation did not increase the ability to photorepair CPD.

We have examined the susceptibility to cyclobutyl pyrimidine dimer (CPD) induction by UVB radiation, the ability to photorepair cyclobutyl pyrimidine dimers (CPDs), and the levels of ultraviolet (UV)-absorbing compounds in various developing stages of leaves in UVB-resistant (Sasanishiki) and -sensitive (Norin 1) rices (Oryza sativa L.). UVB produces more CPDs in the second, third and fourth fully expanded leaves than in the fifth and sixth fully expanded leaves. In leaves in which the levels of UV-absorbing compounds are lower, more CPDs are produced. The ability to photorepair CPDs in fully expanded leaves of different stages of each cultivar remains approximately constant, although the photorepair capacity in the fourth leaf rapidly increases from leaf emergence until full expansion. The photorepair capacity is lower in Norin 1 than that in Sasanishiki in all the leaf stages or different aged leaves. These results therefore suggest that rice plants could be more susceptible to CPD induction at the stage when the second, third or fourth leaves are developing, and UVB sensitivity correlates with low CPD photorepair.

Parental cultivars (Oryza sativa L. Japanese lowland cultivars Sasanishiki and Norin 1), F2 plants, and F3 Iines were grown under visible light with or without supplemental UV-B radiation in a phytotron, to determine whether the accumulation of UV-absorbing compounds plays a role in the difference in UV-B resistance between these rice cultivars. The level of UV-absorbance per unit leaf area increased with the UV-B treatuent in all the leaves of these two rice cultivars, but was higher in Sasanishiki than in Norin 1 irrespective of UV-B treatment. An analysis by thin-layer chromatography and the UV-absorption spectra of the extracts did not show any qualitative difference in UV-absorbing compounds in leaf tissues between these two cultivars. There was a significant positive correlation between the level of UV-absorbance per unit leaf area and the fresh weight of aerial parts in the F2 plants exposed to UV-B radiation. Furthermore, the level of UV-absorbance per unit leaf area tended to be higher in the F3 Iines resistant to UV-B radiation than in those sensitive to such radiation regardless of whether these F3 lines were exposed to supplemental UV-B radiation. These results suggest that the differences in UV-B resistance between Sasanishiki and Norin 1 are associated with genetic differences in the level of accumulation of UV-B absorbing compounds in leaf tissues. Based on these results we suggest that the accumulation of UV-B absorbing compounds is an important factor contributing to UV-B resistance in rice cultivars. Since the accumulation of UV-absorbing compounds in leaves appears to minimize the latent negative impact of UV-B radiation on rice production, it may be a useful character to select in ordinary breeding.

Repair of cyclobutyl pyrimidine dimers (CPDs) in DNA is essential in most organisms to prevent biological damage by ultraviolet (UV) light. In higher plants tested thus far, UV-sensitive strains had higher initial damage levels or deficient repair of nondimer DNA lesions but normal CPD repair. This suggested that CPDs might not be important for biological lesions. The photosynthetic apparatus has also been proposed as a critical target. We have analyzed CPD induction and repair in the UV-sensitive rice (Oryza sativa L.) cultivar Norin 1 and its close relative UV-resistant Sasanishiki using alkaline agarose gel electrophoresis. Norin 1 is deficient in cyclobutyl pyrimidine dimer photoreactivation and excision; thus, UV sensitivity correlates with deficient dimer repair.

The effects of supplementary UV-B radiation (290-320 nm, +UV-B) on the development of wilt disease of spinach (Spinacia oleracea) caused by the fungus Fusarium oxysporum f. sp. spinaciae were investigated under natural light condition in a glasshouse. The disease incidence in these plants greatly increased at low inoculum concentrations in soil inoculation experiments compared with plants without UV-B supplementation (-UV-B). In needle inoculation experiments with F. oxysporum f. sp. spinaciae, wilt occurred in plants under +UV-B as severely as in soil inoculation experiments. F. oxysporum f. sp. spinaciae was isolated from the roots of all wilted plants, as well as some symptomless plants in both soil inoculation and needle inoculation experiments. Re-isolation frequency under +UV-B was ca. 3.5 times higher than that of plants under -UV-B during the first three days of growth under UV-B supplementation. However, the re-isolation frequency in inoculated plants in both experiments irrespective of UV-B supplementation was over 80% by the end of both experiments. The results indicated that supplementary UV-B radiation incited disease development.

The effects of UVB radiation on the growth of two cultivars of Japanese lowland rice {Oryza sativa L.), Sasanishiki and Norin 1, were examined in a phytotron. Supplementation of visible radiation with UVB radiation reduced plant length, tiller number, the fresh and dry weights of the aboveground parts of plants, and the amounts of total leaf nitrogen, chlorophyll, soluble protein and ribulose-l,S-bisphosphate carboxylase/oxygenase (Rubisco) in the eighth leaf, the youngest fully expanded leaf. By contrast, UVB radiation significantly increased the accumulation of UV-absorbing compounds. There was a difference between the two cultivars in the resistance to the effects of UVB radiation. The reduction in the amounts of Rubisco was smaller in Sasanishiki, while the increase in the accumulation of UV-absorbing compounds was greater in Sasanishiki. Parameters of plant growth, with the exception of the amount of Rubisco, decreased in direct proportion to decreases in total leaf nitrogen in plants grown under lower or higher doses of UVB radiation. However, the decrease in the Rubisco content of Norin 1 grown under a high dose of UVB radiation was exceptionally marked, and was not observed similarly in Sasanishiki. These results suggest that the remarkable reduction in Rubisco content in Norin 1 might have been due to the specific effects of UVB radiation. It is also suggested that the difference between cultivars in the resistance to UVB radiation might be due to the differences in the levels of Rubisco and in UV-absorbing compounds that are induced by UVB radiation.

The effects of UV-B radiation (290–320 nm) on development of damping-off of spinach (Spinacia oleracea) caused by the fungusFusarium oxysporum were examined in a growth cabinet. The incidence of disease greatly increased when experimental plants were grown in visible radiation with supplementary UV-B radiation. This increase was suppressed by increasing the irradiation of visible radiation.Fusarium oxysporum was isolated from the roots of all damping-off plants and the roots of some unwilted plants, indicating that spinach infected with the pathogen did not necessarily suffer from damping-off in 15d. Supplementary UV-B radiation suppressed the increase in growth components such as the number of leaves, the plant height and the fresh weight of aboveground plant parts, but did not affect the fresh weight of roots. The ratio of the number of plants infected with pathogen to the total number of plants was over 80% irrespective of light conditions. It was suggested that the defense response of spinach to this pathogen was greatly influenced by the physiological state of aboveground plant parts resulting from supplementary UV-B radiation.

Two types of fungi based on the light required for induction of conidiophore formation are known as ‘photo-induced’ and ‘non-photo-induced’ sporulators. Conidial development in a ‘photo-induced’ strain of Bipolaris oryzae (formerly, Helminthosporium oryzae) is controlled by antagonistic action of blue and near-UV radiation mediated through the ‘mycochrome’ system at two developmental stages: conidiophore induction and conidiophore maturation. Otherwise, conidial development in a ‘non-photo-induced’ strain of B. oryzae, unlike that in the ‘photo-induced’ strain, is controlled by antagonistic action of blue and near-UV radiation at the conidiophore maturation stage alone. In this study, we collected fungi, B. oryzae, from brown lesion spots on rice leaves cultivated in paddy fields, and investigated the possible existence of a new ecotype different from known ‘photo-induced’ and ‘non-photo-induced’ sporulators. We also investigated the sensitivity of conidial development of each isolate to blue and near-UV radiation. The isolates were identified as B. oryzae by morphology of hyphae and conidia and the symptomatic brown lesion spots on rice leaves caused by each isolate. There was no distinction between ‘photo-induced’ and ‘non-photo-induced’ strains on rDNA base. Moreover, the existence of a new type of ‘non-photo-induced’ sporulator different from the one presently known was confirmed. In this newly identified sporulator, conidial development was not affected by light irradiation. Among 153 isolates, one isolate belonged to a previously known ‘non-photo-induced’ sporulator, five isolates belonged to a new ‘non-photo-induced’ sporulator ecotype, and over 95% of the isolates were “photo-induced” sporulators. Among ‘photo-induced’ strains, there were several subtypes having different sensitivities to near-UV radiation required for inducing conidiophore formation.

Genetic analysis of resistance to the inhibitory effects of UV radiation on growth of rice (Oryza sativa L.) cultivars was carried out. Some experimental plants were grown in visible radiation supplemented with UV radiation containing a large amount of UV-B and a small amount of UV-C in a phytotron, while others were grown without UV radiation. The degree of resistance to UV radiation was estimated in terms of the degree of reduction caused by supplemental UV radiation in the fresh weight of the aboveground plant parts and the chlorophyll content per unit fresh weight. Fresh weight and chlorophyll content in F2 plants generated by reciprocally crossing cv. Sasanishiki, a cultivar more resistant to UV radiation, and Norin 1, a cultivar less resistant to such radiation exhibited a normal frequency distribution. The heritabilities of these two properties in F2 plants were low under conditions of non-supplemental UV radiation. Under elevated UV radiation, the F2 population shifted to the lower range of fresh weight and chlorophyll content, and the means were close to those of Norin 1. The heritabilities of these two properties were the same in the reciprocal crosses, indicating that maternal inheritance was not involved. Inheritance of chlorophyll content per unit fresh weight was further determined in F3 lines generated by self-fertilizing F2 plants of Sasanishiki and Norin 1. The results showed that the F3 population was segregated into three genotypes, namely, resistant homozygotes, segregated heterozygotes and sensitive homozygotes, with a ratio of 1:65:16. It was thus evident that the resistance to the inhibitory effect of elevated UV radiation in these rice plants was controlled by recessive polygenes.

Flowering response and plant form of photomorphogenic mutants (hy1, hy2, hy3, hy4 and hy5) of Arabidopsis thaliana (L.), a long-day plant, were examined in long and short days. There were only slight differences among genotypes including Landsberg wild type with respect to the flowering time under long days. The effect of 1 h light-(night)-breaks of far-red, red, blue and white light given in the middle of the dark period of plants grown under short days, was studied. Effects of far-red light applied at the end or the beginning of the main photoperiod on flowering and plant form were also examined. The light-breaks with all the above mentioned light qualities promoted floral initiation of all the genotypes including the wild type in terms of both the flowering time and the number of rosette leaves. In general, far-red light was most effective. It is possible to classify the hy-mutants into 3 groups by their responses to light-breaks under short day conditions: (a) Mutants hy2 and hy3, which have a reduced number of rosette leaves, and flower early. Red light is as effective as far-red light. The wavelength of light-breaks is relatively unimportant for flowering response. (b) Mutants hy4, hy5 and Landsberg wild type, which have a greater number of rosette leaves, and flower relatively late. The effectiveness of light-breaks is in the following order, far-red, blue, and red light, which is in reverse order to the transformation of phytochrome to the Pfr form. (c) Mutant hy1, which behaves anomalously with respect to relations between flowering time and number of rosette leaves; late flowering with reduced number of rosette leaves. Red, blue and far-red light are effective, but white light is ineffective for reducing the number of rosette leaves. When far-red light was given in the middle of the night or at the end of the main photoperiod, it markedly reduced the number of rosette leaves compared to those grown under short days for all the genotypes, while when applied at the beginning of the main photoperiod far-red light did not affect the number of rosette leaves. Different effects on the plant form dependent on the time of treatment with far-red light-breaks are also discussed.

Application of an inhibitor of gibberellin biosynthesis alleviated the UV-B injury in rice seedlings when tested for chlorophyll content per unit fresh and dry weight. A genetic study was conducted by crossing Tan-ginbozu, a gibberellin-deficient mutant, against two floating rice cultivars that presumably contain large amounts of endogenous gibberellins. The heritabilities of plant height, and chlorophyll content per unit fresh and dry weight were high in the F2 plants in non UV-B irradiated control populations. When those plants were treated with UV-B irradiation, the F2 population tended to shift to the lower range in the chlorophyll content per unit weight, and the value of heritability fluctuated depending on the cross and characters measured. Selection of plants that are highly resistant to UV-B irradiation can be made possible by finding the range of UV-B energy that increases the difference between sensitive and resistant, while also increasing the value of the heritability.

Rice cultivars vary widely in their sensitivity to ultraviolet B (UVB) and this has been correlated with cyclobutane pyrimidine dimer (CPD) photolyase mutations that alter the structure/function of this photorepair enzyme. Here, we tested whether CPD photolyase function determines the UVB sensitivity of rice (Oryza sativa) by generating transgenic rice plants bearing the CPD photolyase gene of the UV-resistant rice cultivar Sasanishiki in the sense orientation (S-B and S-C lines) or the antisense orientation (AS-D line). The S-B and S-C plants had 5.1- and 45.7-fold higher CPD photolyase activities than the wild-type, respectively, were significantly more resistant to UVB-induced growth damage, and maintained significantly lower CPD levels in their leaves during growth under elevated UVB radiation. Conversely, the AS-D plant had little photolyase activity, was severely damaged by elevated UVB radiation, and maintained higher CPD levels in its leaves during growth under UVB radiation. Notably, the S-C plant was not more resistant to UVB-induced growth inhibition than the S-B plant, even though it had much higher CPD photolyase activity. These results strongly indicate that UVB-induced CPDs are one of principal causes of UVB-induced growth inhibition in rice plants grown under supplementary UVB radiation, and that increasing CPD photolyase activity can significantly alleviate UVB-caused growth inhibition in rice. However, further protection from UVB-induced damage may require the genetic enhancement of other systems as well.

The inheritance of light dependence for conidial development inBipolaris oryzae was analyzed using single-ascospore isolates. When a photo-induced strain was crossed with another photo-induced strain, only photo-induced progeny were produced. When a photo-induced strain was crossed with a non-photo-induced (I) strain, photo-induced and nonphoto-induced (I) progeny were produced in a ratio of 1∶1. However, when a photo-induced strain was crossed with a non-photo-induced (II) strain, a non-photo-induced (I) progeny were formed in addition to parental types. It was suggested that genes for photo-control of conidiophore induction and genes for photo-control of conidiophore maturation are located on the same chromosome.

Sensitivity to ultraviolet-B (UVB) radiation (280-320 nm) varies widely among rice cultivars. We previously indicated that UV-resistant rice cultivars are better able to repair cyclobutane pyrimidine dimers (CPDs) through photorepair than are UV-sensitive cultivars. In this paper, we report that UVB sensitivity in rice, in part, is the result of defective CPD photolyase alleles. Surjamkhi (indica) exhibited greater sensitivity to UVB radiation and was more deficient in CPD photorepair ability compared with UV-resistant Sasanishiki (japonica). The deficiency in CPD photorepair in Surjamkhi resulted from changes in two nucleotides at positions 377 and 888 in the photolyase gene, causing alterations of two deduced amino acids at positions 126 and 296 in the photolyase enzyme. A linkage analysis in populations derived from Surjamkhi and Sasanishiki showed that UVB sensitivity is a quantitative inherited trait and that the CPD photolyase locus is tightly linked with a quantitative trait locus that explains a major portion of the genetic variation for this trait. These results suggest that spontaneously occurring mutations in the CPD photolyase gene cause different degrees of sensitivity to UVB in rice, and that the resistance of rice to UVB radiation could be increased by increasing the photolyase function through conventional breeding or bioengineering.

There is a cultivar difference in the response to ultraviolet-B (UVB: 280-320 nm) in rice (Oryza sativa L.). Among Japanese lowland rice cultivars, Sasanishiki, a leading Japanese rice cultivar, is resistant to the damaging effects of UVB while Norin 1, a close relative, is less resistant. We found previously that Norin 1 was deficient in cyclobutane pyrimidine dimer (CPD) photorepair ability and suggested that the UVB sensitivity in rice depends largely on CPD photorepair ability. In order to verify that suggestion, we examined the correlation between UVB sensitivity and CPD photolyase activity in 17 rice cultivars of progenitors and relatives in breeding of UV-resistant Sasanishiki and UV-sensitive Norin 1. The amino acid at position 126 of the deduced amino acid sequence of CPD photolyase in cultivars including such as Norin 1 was found to be arginine, the CPD photolyase activities of which were lower. The amino acid at that position in cultivars including such as Sasanishiki was glutamine. Furthermore, cultivars more resistant to UVB were found to exhibit higher photolyase activities than less resistant cultivars. These results emphasize that single amino acid alteration from glutamine to arginine leads to a deficit of CPD photolyase activity and that CPD photolyase activity is one of the main factors determining UVB sensitivity in rice.

Among Indica rice cultivars (Oryza sativa L. cvs.) that belong to the aus ecotype from the tropical Bengal region, where the amount of ultraviolet-B (UV-B) radiation in the solar radiation is relatively great, Marich-bati cultivar has exhibited resistance to UV-B radiation, while Surjamkhi cultivar appeared to be less resistant. We have examined the susceptibility to cyclobutane pyrimidine dimer (CPD) induction by UV-B radiation and the ability to photorepair CPDs using these two cultivars. UV-B radiation produced similar dimer levels in the leaves of the two cultivars. In contrast, the ability to photorepair CPDs in the UV-sensitive Surjamkhi cultivar was lower than that in the UV-resistant Marich-bati cultivar. These results were similar to our previous data, namely, that a UV-sensitive Japanese rice cultivar (Oryza sativa L. cv. Norin 1) cultivated in the moderate climate of Japan is deficient in its ability to photorepair CPDs. Thus, these results suggest that a strong correlation exists between the sensitivity to UV-B and the photorepair deficiency, and that a low ability in CPD photorepair may be a principal factor in determining the UV-B sensitivity in rice plants.

Verticillium agaricinum (Link) Corda, grown in a yeast extract-sucrose medium, conidiated abundantly in darkness after irradiation with near ultraviolet (290–400 nm) for 15 min or blue light (400–550 nm) for 60 min. Few conidia were formed in total darkness. Exposure to 30 min of near ultraviolet light suppressed conidiation. Conidiation was also suppressed by phosphate in excess of 10−4M irrespective of light condition. After irradiation with near ultraviolet light for more than 30 min, there was a cessation of growth and a change in colony color from yellow to reddish. The color does not appear to be due to a carotenoid because the colonies turned from red to yellow when covered with acid. At pH lower than 6.0 the pigment has an absorption maximum around 390 nm, whereas at higher pH it is around 540 nm. Thus, it appears that irradiation of V. agaricinum with near ultraviolet may cause an increase in pH, which in turn produces the change of colony color from yellow to reddish.

— When dark-grown colonies were exposed to near-UV light, conidiophore formation was induced, and conidia developed during a subsequent dark period. Simultaneous exposure to near-UV and blue light inhibited induction of conidiation. The inductive effect of near-UV light irradiation was greatly enhanced by treating colonies with low temperatures for 4 h during the 2nd-6th hour of incubation in the dark following inductive irradiation: the enhancement was greatest at 21°C. The extent of inhibition by blue light increased with the temperature between 10 and 28°C. This diminution by low temperature was greatest when colonies previously kept at a low temperature were exposed to inductive irradiation: the longer the duration between inductive irradiation and temperature treatment, the lower the diminution. Higher fluences of blue light were required to suppress conidial induction at lower temperature. Thus, it is evident that the inductive effect of near-UV light irradiation on conidiation and the suppressive effect of blue light irradiation are each responsive to different temperatures applied at different times.

Further studies have been made on the blue and near ultraviolet-reversible photoreaction in a cell-free system of the fungus Alternaria tomato, prepared by two-layer sucrose density gradient centrifugation. A light-minus-dark difference spectrum of the supernatant fraction showed a dip in the near ultraviolet region, mainly at 280 nm, when irradiated with near ultraviolet light which could not be reversed by following the initial irradiation with blue light. The difference spectrum of the dialyzed particulate fraction, located in the 68% sucrose band, showed a peak absorbance in the blue region, mainly at 400 nm, when irradiated with near ultraviolet light, and also could not be reversed by blue light. When the particulate fraction plus the supernatant fraction was irradiated with near ultraviolet light, the increment rate of peak absorbance in the blue region increased compared to that of the particulate fraction lacking the supernatant, and both peak absorbance in the blue region and the dip in the near ultraviolet region were reversed by irradiation with blue light following the application of near ultraviolet light. Thus, it was concluded that two photoresponsive systems are involved in blue and near ultraviolet reversible photoreaction on an intracellular particulate fraction.

Coniditation the fungus, Alternaria cichorii Nattras (IMI 225641), was repeatedly induced and inhibited by alternating doses of near ultraviolet and blue radiation. The final response largely depended on the last kind of radiation administered. When the final radiation was near ultraviolet, conidiophore formation was induced, and conidia were then produced in the subsequent dark period; when it was blue, the induction of conidiation was suppressed. The inductive effect of near ultraviolet was promoted by anaerobiosis, but the reversing effect by blue radiation was not influenced by molecular oxygen. Thus, the “mycochrome” system apprears to control induction of conidiation in Alternaria cichorii, and oxidation-reduction processes may be involved in its function.

Near ultraviolet irradiation and its significance to the photoresponsive system “mycochrome” (involved in blue and near ultraviolet reversible photoreaction) in Alternaria tomato were studied. Near ultraviolet irradiation caused photoreduction of both PB (blue absorbing pigment in mycochrome) and cytochrome c mediated through PNUV (near ultraviolet absorbing pigment in mycochrome). Both PNUV mediated photoreductions of PB and cytochrome c were larger under anaerobic conditions than aerobic conditions. Both photoreductions under aerobic conditions could be inhibited by superoxide dismutase; consequently, it is considered that a superoxide anion was produced when PNUV was exposed to near ultraviolet light under aerobic conditions which also reduced PB and cytochrome c. Under anaerobic conditions, some photo-product resulting from the cooperation of PNUV and near ultraviolet light (with the exception of a superoxide anion) may directly reduce either PB or cytochrome c, since these photoreductions could not b...

Photosporulation of Alternaria tomato was very little influenced by polyoxin at concentrations below 10 μg/ml, although hyphae and conidia were morphologically affected. The hyphae grown in the presence of polyoxin were easily disrupted.

Conidiation in A. cichorii Nattras is reversibly stimulated by near UV radiation (NUV .apprx. 313 nm) and inhibited by blue light (.apprx. 450 nm) and apparently is a mycochrome-mediated process. After induction with plane-polarized NUV, blue light polarized perpendicularly to the NUV was more effective in counteracting the induction than was blue light polarized parallel to the NUV. Both the blue-absorbing component (presumably a flavo-protein) and the PNUV (NUV absorbing moiety) of the mycochrome system are membrane-bound. The transition moment associated with blue light absorption in the presumed flavoprotein forms an angle of at least 53.degree. with the transition moment associated with NUV absorption in PNUV.

Conidiophore formation in Helminthosporium oryzae V. Breda de Haan (ATCC 38851) was induced by near-ultraviolet radiation, and the conidia developed in subsequent darkness. With regard to repetitive near-ultraviolet light pulses, more fluence was needed to induce the same level of conidiation as that induced by continuous near-ultraviolet radiation. With a lengthening of the duration of the dark-intervals (50 ms to 10 s) between 50-ms light pulses, much more fluence was required. On the other hand, blue radiation applied just before near-ultraviolet radiation suppressed the induction of conidiation. When preirradiated with repetitive blue light pulses, the fluence needed to suppress induction of conidiation to the same level as that of continuous blue radiation was somewhat less than that for continuous blue radiation. The longer the dark-interval duration (50 ms to 10 s) between light pulses, the less the fluence of blue radiation required. These results indicate that not only the photoreaction but also the dark reaction was very important for the accumulation of photo stimuli for both the blue and the near-ultraviolet light responses; however, they were kinetically quite different from each other.

Lemna paucicostata Hegelm. T-101, a short-day plant, flowers when plants preirradiated with red light (R) for 24 h are subjected to inductive darkness for 72 h followed by two short-day cycles (6 h R+ 18 h dark). However, flowering is inhibited by blue-or far-red-light pulses applied at the beginning of the inductive dark period. These inhibitory light effects are fully reversible by a R pulse. The action spectra for the inhibitory light effect and for its reversal show that the light pulses act exclusively through phytochrome. It is concluded that a low level of Pfr at the beginning of the inductive dark period prevents flowering.