The filamentous fungus Alternaria alternata produces melanin, a black pigment, from acetate via 1,8-dihydroxynaphthalene. To isolate a fungal gene required for melanin biosynthesis, we transformed an A. alternata Brm1- (light brown) mutant with the DNA of a wild-type strain genomic library constructed by use of a cosmid carrying the hygromycin B phosphotransferase gene. When hygromycin B-resistant transformants were screened for melanin production, 1 of 1,363 transformants appeared to regain melanin production, as judged by black pigmentation of the cultured mycelia. The cosmid, named pMBR1, was recovered by packaging nuclear DNA of the melanin-producing transformant into lambda phage. The gene on pMBR1 that enables the Brm1- mutant to produce melanin was designated BRM1. In addition to the BRM1 gene, pMBR1 was found to carry two more genes involved in melanin biosynthesis. These two genes, designated ALM and BRM2, transformed A. alternata Alm- (albino) and Brm2- (brown) mutants, respectively, to the wild-type phenotype. The three genes are located within a ca. 30-kb genomic region in the order ALM-BRM1-BRM2. Analysis of the gene transcripts indicated approximate sizes of 7.2, 4.0, and 0.9 kb for ALM, BRM1, and BRM2, respectively. The BRM1 and BRM2 transcripts are generated from the same strand, but the ALM transcript is generated from the opposite strand. The three mRNA species accumulate in cultured mycelia of the wild-type strain synchronously with mycelial melanization. The essential roles of the three genes in melanin biosynthesis were confirmed by transformation-mediated gene disruption experiments.

The Improved Limb Atmospheric Spectrometer‐II (ILAS‐II) monitored components associated with Polar ozone depletion. ILAS‐II was on board the Advanced Earth Observing Satellite‐II (ADEOS‐II, “Midori‐II”), which was successfully launched on 14 December 2002 from the Tanegashima Space Center of the Japan Aerospace Exploration Agency (JAXA). ILAS‐II used a solar occultation technique to measure vertical profiles of ozone (O3), nitric acid (HNO3), nitrogen dioxide (NO2), nitrous oxide (N2O), methane (CH4), water vapor (H2O), chlorine nitrate (ClONO2), dinitrogen pentoxide (N2O5), CFC‐11, CFC‐12 and aerosol extinction coefficients at high latitudes in both the Northern and Southern hemispheres. ILAS‐II included Sun‐tracking optics and four spectrometers, a Sun‐edge sensor, and electronics. The four spectrometers measured in the infrared (channel 1) between 6.21 and 11.76 μm, in the midinfrared (channel 2) between 3.0 and 5.7 μm, at high resolution in the infrared (channel 3) between 12.78 and 12.85 μm, and in the visible (channel 4) between 753 and 784 nm. The vertical height of the entrance slit was 1 km at the tangent point. A Sun‐edge sensor accurately registered tangent height. After an initial check of the instruments, ILAS‐II recorded routine measurements for about 7 months, from 2 April 2003 to 24 October 2003, a period that included the formation and collapse of an Antarctic ozone hole in 2003 that was one of the largest in history. All of the ILAS‐II data were processed using the version 1.4 data‐processing algorithm. Validation analyses show promising results for some ILAS‐II measurement species, which can be used to elucidate mechanisms of Polar ozone depletion. Studies are ongoing on ozone depletion, on the formation mechanisms of Polar stratospheric clouds, on denitrification, and on air mass descent. A state‐of‐the‐art data retrieval algorithm that is currently being developed will yield more sophisticated data sets from the ILAS‐II data in the near future.

The phytopathogenic fungi Magnaporthe grisea and Alternaria alternata produce melanin via the polyketide biosynthesis, and both fungi form melanized colonies. However, the site of melanin deposition and the role of melanin in pathogenicity differ between these two fungi. M. grisea accumulates melanin in appressoria, and their melanization is essential for host penetration. On the other hand, A. alternata produces colorless appressoria, and melanin is not relevant to host penetration. We examined whether the melanin biosynthesis genes of A. alternata could complement the melanin-deficient mutations of M. grisea. Melanin-deficient, nonpathogenic mutants of M. grisea, albino (Alb-), rosy (Rsy-), and buff (Buf-), were successfully transformed with a cosmid clone pMRB1 that carries melanin biosynthesis genes ALM, BRM1, and BRM2 of A. alternata. This transformation restored the melanin synthesis of the Alb- and Buf- mutants, but not that of the Rsy- mutant. The melanin-restored transformants regained mycelial melanization, appressorium melanization, and pathogenicity to rice. Further, transformation of Alb- and Buf- mutants with subcloned ALM and BRM2 genes, respectively, also produced melanin-restored transformants. These results indicate that the Alternaria genes ALM and BRM2 can restore pathogenicity to the mutants Alb- and Buf-, respectively, due to their function during appressorium development in M. grisea.

Two new Pseudomonas species which were isolated from rice paddy and clinical specimens (groups Ve-2 and Ve-1) are described. Strains of Pseudomonas oryzihabitans sp. nov. are yellow-pigmented, oxidase-negative, nonsporeforming, gram-negative, polarly monotrichously flagellated, rod-shaped organisms with deoxyribonucleic acid base compositions ranging from 63.9 to 65.6 mol% guanine plus cytosine, ubiquinone Q-9, major cellular fatty acids consisting of In the course of a study of the microflora of rice paddies, a large number of yellow-pigmented, gram-negative bacteria were isolated. Some of these organisms were identified as "Pseudomonas lacunogenes," which was studied by Goresline (7), and some characteristics of our isolates have been published previously (12). When we published the characteristics of our strains of "P. lacunogenes," the original strains which Goresline used for the description of this species and reference strains were not available from any culture collection or other sources. They are still unavailable. The strains of "P. lacunogenes" were a major component of the bacterial flora of normal rice paddies (11). However, this name is not on the Approved Lists of Bacterial Names (26). Recently, we isolated new strains similar to "P. lacunogenes" from normal rice paddies. These cultures were oxidase negative and resembled cultures that Weaver et al. (31), Tatum et al. (30), and Gilardi et al. (6) recognized as groups Ve-1 and Ve-2 of yellow-pigmented, oxidative, oxidase-negative, gram-negative, polarly flagellated, rodshaped, bacteria. Therefore, we attempted to identify our isolates from normal rice paddies by using authentic strains representative of groups Ve-1 and Ve-2.In this paper we describe two new species, Pseudomonas oryzihabitans sp. nov., which was isolated from normal rice paddies and clinical specimens (group Ve-2), and Pseudomonus luteola sp. nov., which was isolated from clinical specimens (group Ve-1). Chemical Research, Wako-shi, Saitama, Japan; NCIB, National Collection of Industrial Bacteria, Torrey Research Station, Aberdeen, United Kingdom; DNA, deoxyribonucleic acid; G+C, guanine plus cytosine; DNase, deoxyribonuclease; ONPG, o-nitrophenyl-P-galactopyranoside. The designations used for ubiquinones indicate the number of isoprene units in a side chain (e.g., ubiquinones Q-8, Q-9, and Q-10, have 8, 9, and 10 isoprene units, respectively). MATERIALS AND METHODSBacterial strains. The strains which we studied are indicated below (names which do not appear on the Approved Lists of Bacterial Names [26] are enclosed in quotation marks)., and KS0904 (= D-2 = AJ 2210) were isolated from normal rice paddies by Iizuka and Komagata (12). Strain KS0905 was isolated from Oryza sativa strain C5444 (grain harvested in 1979), strain KS0906 was isolated from 0. sativa strain C8895 (grain harvested in 1979), strain KS0907 was isolated from 0. sativa strain (28896 (grain harvested in 1979), strain KS0908 was isolated from Oryza breviligulata strain W1064 (grain harvested in 1977), strain KS09...

The Improved Limb Atmospheric Spectrometer (ILAS) was a satellite‐based solar occultation sensor that was developed by the Environment Agency of Japan (EA) to monitor and study the stratospheric ozone layer. This paper describes the characteristics of the ILAS instrument and its performance in orbit. ILAS measured the vertical distribution of ozone, nitric acid, nitrogen dioxide, nitrous oxide, methane, water vapor, temperature, pressure, and aerosol extinction coefficients at 1.6‐km vertical resolution. ILAS was equipped with two spectrometers: an infrared (IR) spectrometer with an uncooled pyroelectric linear array detector to sense between 6.21 and 11.76 μm and a visible spectrometer to monitor 753–784 nm. In addition, a Sun‐edge sensor (SES) assigned the tangent height of the instantaneous field‐of‐view (IFOV). A two‐axis gimbals control system on ILAS used two Sun position sensors to track the center of brightness of the Sun during occultation measurements. Before launch onboard the Advanced Earth Observing Satellite (ADEOS), the performance of ILAS was checked on the ground using several methods, including gas‐cell measurements, time response measurements, Sun‐tracking tests, and hollow‐cathode lamp measurements. After the launch of ADEOS on 17 August 1996, ILAS functioned successfully for 8 months of routine operation, from 30 October 1996 to 30 June 1997, collecting more than 6700 solar occultation measurements, after which time the satellite failed due to a failure in a solar paddle. The time delay response of the IR channel was characterized using stepwise IR input. Instrument functions of the ILAS IR and visible spectrometers were determined by combining theoretical optical calculations, experimental measurements using a gas‐cell before launch, and in‐orbit data. The signal‐to‐noise ratio (SNR) of each element in the IR channel was estimated to be 400–1200. In the visible channel, it was 1600–1800 for a 100% direct Sun signal. At sunset occultation, ILAS was able to track the Sun below a tangent height of 10 km in some cases. The method of determining the solar edges from the SES data worked correctly, giving adequate tangent height information for observations. Output signal levels of the SES, visible channel, and IR channel showed slight degradation during the period that ILAS was operational, which is attributed to space‐borne contaminants. However, changes in absolute signal levels do not affect data retrieval, because the solar occultation technique was self‐calibrating. Overall, ILAS worked as designed during its operation in orbit and gathered valuable data for ozone layer studies.