SummaryThe role of phytochrome B2 (phyB2) in the control of photomorphogenesis in tomato (Solanum lycopersicum L.) has been investigated using recently isolated mutants carrying lesions in the PHYB2 gene. The physiological interactions of phytochrome A (phyA), phytochrome B1 (phyB1) and phyB2 have also been explored, using an isogenic series of all possible mutant combinations and several different phenotypic characteristics. The loss of phyB2 had a negligible effect on the development of white-lightgrown wild-type or phyA-de®cient plants, but substantially enhanced the elongated pale phenotype of the phyB1 mutant. This redundancy was also seen in the control of de-etiolation under continuous red light (R), where the loss of phyB2 had no detectable effect in the presence of phyB1. Under continuous R, phyA action was largely independent of phyB1 and phyB2 in terms of the control of hypocotyl elongation, but antagonized the effects of phyB1 in the control of anthocyanin synthesis, indicating that photoreceptors may interact differently to control different traits. Irradiance response curves for anthocyanin synthesis revealed that phyB1 and phyB2 together mediate all the detectable response to high-irradiance R, and, surprisingly, that the phyA-dependent low-irradiance component is also strongly reduced in the phyB1 phyB2 double mutant. This is not associated with a reduction in phyA protein content or responsiveness to continuous far-red light (FR), suggesting that phyB1 and phyB2 speci®cally in¯uence phyA activity under low-irradiance R. Finally, the phyA phyB1 phyB2 triple mutant showed strong residual responsiveness to supplementary daytime FR, indicating that at least one of the two remaining phytochromes plays a signi®cant role in tomato photomorphogenesis.
SummarySeveral novel allelic groups of tomato (Solanum lycopersicum L.) mutants with impaired photomorphogenesis have been identi®ed after g-ray mutagenesis of phyA phyB1 double-mutant seed. Recessive mutants in one allelic group are characterized by retarded hook opening, increased hypocotyl elongation and reduced hypocotyl chlorophyll content under white light (WL). These mutants showed a speci®c impairment in response to blue light (BL) resulting from lesions in the gene encoding the BL receptor cryptochrome 1 (cry1). Phytochrome A and cry1 are identi®ed as the major photoreceptors mediating BL-induced de-etiolation in tomato, and act under low and high irradiances, respectively. Phytochromes B1 and B2 also contribute to BL sensing, and the relative contribution of each of these four photoreceptors differs according to the light conditions and the speci®c process examined. Development of the phyA phyB1 phyB2 cry1 quadruple mutant under WL is severely impaired, and seedlings die before¯owering. The quadruple mutant is essentially blind to BL, but experiments employing simultaneous irradiation with BL and red light suggest that an additional non-phytochrome photoreceptor may be active under short daily BL exposures. In addition to effects on de-etiolation, cry1 is active in older, WL-grown plants, and in¯uences stem elongation, apical dominance, and the chlorophyll content of leaves and fruit. These results provide the ®rst mutant-based characterization of cry1 in a plant species other than Arabidopsis.
Genebank collection databases can be used for ecogeographical studies under the assumption that the accessions are a geographically unbiased sample. We evaluated the representativeness of a collection of wild potatoes from Bolivia and defined and assessed four types of bias: species, species-area, hotspot, and infrastructure. Species bias is the sampling of some species more often than others. Species-area bias is a sampling that is disproportionate to the total area in which a species is found. Hotspot bias is the disproportionate sampling of areas with high levels of diversity. Infrastructure bias is the disproportionate sampling of areas near roads and towns. Each of these biases is present in the Bolivian wild potato collection. The infrastructure bias was strong: 60% of all wild potato accessions were collected within 2 km of a road, as opposed to 22%, if collections had been made randomly. This analysis can serve as a guide for future collecting trips. It can also provide baseline information for the application of genebank data in studies based on geographic information systems. Evaluación de la Representatividad Geográfica de las Colecciones de Bancos de Genes: El Caso de las Papas Silvestres BolivianasResumen: Las bases de datos de los bancos de germoplasma pueden ser usadas para estudios ecogeográficos bajo el supuesto que las entradas constituyen una muestra geográficamente imparcial. Evaluamos la representatividad geográfica de una colección de papas silvestres de Bolivia y definimos y evaluamos cuatro tipos de sesgos: sesgos de especie, de especie-área, de áreas con gran diversidad (hotspot), y de infraestructura. El sesgo de especie implica el muestrear más algunas especies que otras. El sesgo de especie-área es un muestreo que es desproporcionado con respecto al total del área en la cual se encuentra una especie. El sesgo de "hotspot" es el muestreo desproporcionado de áreas con niveles altos de diversidad. El sesgo por infraestructura es aquel muestreo desproporcionado de áreas cercanas a carreteras y pueblos. Cada uno de estos sesgos se presenta en la colección de papas silvestres de Bolivia. El sesgo por infraestructura fue muy elevado: 60% de todas las entradas de papas silvestres fueron colectados dentro de un radio de 2 km de distancia de las carreteras, cuando se debería esperar un 22% si las colectas se hubieran hecho de manera aleatoria. Este análisis puede servir como guía para futuras exploraciones y proporciona una base para la aplicación de los datos de bancos de genes en estudios basados en sistemas de información geográfica. *
Glycosyltransferases are involved in the biosynthesis of lipid-linked N-glycans. Here, we identify and characterize a mannosyltransferase gene from Arabidopsis thaliana, which is the functional homolog of the ALG3 (Dol-P-Man:Man 5 GlcNAc 2 -PP-Dol a1,3-mannosyl transferase) gene in yeast. The At ALG3 protein can complement a Dalg3 yeast mutant and is localized to the endoplasmic reticulum in yeast and in plants. A homozygous T-DNA insertion mutant, alg3-2, was identified in Arabidopsis with residual levels of wild-type ALG3, derived from incidental splicing of the 11th intron carrying the T-DNAs. N-glycan analysis of alg3-2 and alg3-2 in the complex-glycan-less mutant background, which lacks N-acetylglucosaminyl-transferase I activity, reveals that when ALG3 activity is strongly reduced, almost all N-glycans transferred to proteins are aberrant, indicating that the Arabidopsis oligosaccharide transferase complex is remarkably substrate tolerant. In alg3-2 plants, the aberrant glycans on glycoproteins are recognized by endogenous mannosidase I and N-acetylglucosaminyltransferase I and efficiently processed into complex-type glycans. Although no high-mannose-type glycoproteins are detected in alg3-2 plants, these plants do not show a growth phenotype under normal growth conditions. However, the glycosylation abnormalities result in activation of marker genes diagnostic of the unfolded protein response.
Abstract— A high‐pigment (hp) mutant, which shows exaggerated phytochrome responses and three other genotypes of Lycopersicon esculenrum Mill. cv. Ailsa Craig: the aurea (au) mutant deficient in the bulk light‐labile phytochrome (PI) pool, the au, hp double mutant, and their isogenic wild type, were used in this study. Measurements of phytochrome destruction in red light (R) revealed that the exaggerated responses of the hp mutant are not caused by a higher absolute phytochrome level or a reduced rate of phytochrome destruction. Fluence‐response relationships for anthocyanin synthesis after a blue‐light pretreatment were studied to test if the hp mutant conveys hypersensitivity to the far‐red light (FR)‐absorbing form of phytochrome (Pfr), i.e. the threshold of Pfr required to initiate the response is lower. The response range for the hp mutant and wild type was identical, although the former exhibited a 6‐fold larger response. Moreover, the kinetics of anthocyanin accumulation in continuous R were similar in the wild‐type and hp‐mutant seedlings, despite the latter accumulating 9‐fold more anthocyanin. Since the properties of phytochrome are the same, the hp mutation appears to affect the state of responsiveness amplification, i.e. the same amount of Pfr leads to a higher response in the hp mutant. We therefore propose that the hp mutation is associated with an amplification step in the phytochrome transduction chain. Escape experiments showed that the anthocyanin synthesis after different light pretreatments terminated with a R pulse was still 50% FR reversible after 4–6 h darkness, indicating that the Pfr pool regulating this response must be relatively stable. However, fluence‐rate response relationships for anthocyanin synthesis and hypocotyl growth induced by a 24‐h irradiation with 451, 539, 649, 693, 704 and 729 nm light showed no or a severely reduced response in the au and au, hp mutants, suggesting the importance of PI in these responses. We therefore propose that the capacity for anthocyanin synthesis (state of responsiveness amplification) could be established by PI, while the anthocyanin synthesis is actually photoregulated via a stable Pfr pool. The Hp gene product is proposed to be an inhibitor of the state of responsiveness amplification for responses controlled by this relatively stable Pfr species.
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