Genetic characterization and high efficiency photosynthesis of an aurea mutant of tobacco

A new tobacco (Nicotiana tabacum) aurea mutant was isolated from the progeny of a selfed variegated tobacco plant. The new mutant is termed Su/su var. Aurea. If the mutant is selfed, the seeds obtained give rise to four types of plants: green seedlings which correspond to the wild type; yellow-green seedlings which correspond to the earlier described Su/su; yellow seedlings which correspond to the new tobacco aurea mutant Su/su var. Aurea; and white lethal seedlings. The frequency ratio of the four phenotypes is 1:1:1:1. It appears that the mutation is due to two independent nuclear factors, su and aur, both of which have to be present in a heterozygous conditions, Su/su Aur/aur, to give rise to the new aurea phenotype. The aurea mutant Su/su var. Aurea has a reduced photosynthetic unit size which is approximately one-eighth of the wild type. Despite its chlorophyll deficiency, the plant grows well and exhibits maximal photosynthetic rates on a chlorophyll basis which are at least seven times higher than those of the green wild type provided the temperature and the light intensities are high enough. In contrast to the earlier described Su/su, the new mutant does not exhibit more photorespiration than the wild type. It appears that the factor aur causes either repression of photorespiration or an increase in the number of functioning photosynthetic units.     

Effects of light quality on the chloroplastic ultrastructure and photosynthetic characteristics of cucumber seedlings

To understand how light quality influences plant photosynthesis, we investigated chloroplastic ultrastructure, chlorophyll fluorescence and photosynthetic parameters, Rubisco and chlorophyll content and photosynthesis-related genes expression in cucumber seedlings exposed to different light qualities: white, red, blue, yellow and green lights with the same photosynthetic photon flux density of 100 μmol m^?2 s^?1. The results revealed that plant growth, CO₂ assimilation rate and chlorophyll content were significantly reduced in the seedlings grown under red, blue, yellow and green lights as compared with those grown under white light, but each monochromatic light played its special role in regulating plant morphogenesis and photosynthesis. Seedling leaves were thickened and slightly curled; Rubisco biosynthesis, expression of the rca, rbcS and rbcL, the maximal photochemical efficiency of PSII (Fv/Fm) and quantum yield of PSII electron transport (Ф_PSII) were all increased in seedlings grown under blue light as compared with those grown under white light. Furthermore, the photosynthetic rate of seedlings grown under blue light was significantly increased, and leaf number and chlorophyll content of seedlings grown under red light were increased as compared with those exposed to other monochromatic lights. On the contrary, the seedlings grown under yellow and green lights were dwarf with the new leaves etiolated. Moreover, photosynthesis, Rubisco biosynthesis and relative gene expression were greatly decreased in seedlings grown under yellow and green light, but chloroplast structural features were less influenced. Interestingly, the Fv/Fm, Ф_PSII value and chlorophyll content of the seedlings grown under green light were much higher than those grown under yellow light.     

GENETIC HISTORY AND GENERAL COMPARISONS OF TWO ALBINO MUTATIONS OF HELIANTHUS ANNUUS

Wallace , Raymond H. (U. Connecticut, Storrs.), and Helen M. Habermann . Genetic history and general comparisons of two albino mutations of Helianthus annuus. Amer. Jour. Bot. 46(3) : 157-162. Illus. 1959.—The genetic history of the progeny of a single ultrasonically-treated seedling of Helianthus annuus L. has been summarized for the 6 generations for which quantitative data are available. A yellow mutation was found in the F₂ generation and later in the F₅, a second, white mutation occurred. Both mutants have been grown to maturity by grafting them onto normal green host plants and they have set viable seeds. Both pigment-deficient conditions are inherited as single recessive factors. These albino strains form chlorophyll during their seedling stages if they are grown at low light intensities. Chlorophyll is destroyed, however, under bright illumination and, once bleached, the capacity for chlorophyll formation appears to be lost. The yellow mutant contains xanthophyll but no traces of carotene have been found. In the white mutant, neither carotene nor xanthophyll have been detected. Flower color in the yellow mutant is normal while the flowers of the white mutant have no apparent pigmentation. The growth pattern of grafted yellow mutants is normal, save for a stiffer and woodier condition and a greater resistance to wilting. These characteristics have also been observed in grafted white mutants. In addition, there is a pronounced reduction in leaf size in the white mutant.     

Chlorophyll reduction in the seed of Brassica napus with a glutamate 1-semialdehyde aminotransferase antisense gene*

Chlorophyll reduction in the seed of Brassica can be achieved by downregulating its synthesis. To reduce chlorophyll synthesis, we have used a cDNA clone of Brassica napus encoding glutamate 1-semialdehyde aminotransferase (GSA-AT) to make an antisense construct for gene manipulation. Antisense glutamate 1-semialdehyde aminotransferase gene (Gsa) expression, directed by a Brassica napin promoter, was targeted specifically to the embryo of the developing seed. Transformants expressing antisense Gsa showed varying degrees of inhibition resulting in a range of chlorophyll reduction in the seeds. Seed growth and development were not affected by reduction of chlorophyll. Seeds from selfed transgenic plants germinated with high efficiency and growth of seedlings was vigorous. Seedlings from T₂ transgenic lines segregated into three distinctive phenotypes: dark green, light green and yellow, indicating the dominant inheritance of Gsa antisense gene. These transgenic lines have provided useful materials for the development of a low chlorophyll seed variety of B. napus.     

CHROMOPLAST ULTRASTRUCTURE AS AFFECTED BY GENES CONTROLLING GRANA RETENTION AND CAROTENOIDS IN FRUITS OF CAPSICUM ANNUUM

Plastids in the fruits of isogenic lines of pepper (Capsicum annuum) were examined by electron microscopy with reference to four genotypes determining the carotenoid composition and the colors red, yellow, brown, and green of the ripe fruit. One gene pair (y⁺/y) influences carotenoid content and the other pair (cl⁺/cl) controls the chlorophyll. The retention of the grana and chlorophyll in the ripe fruits of the brown and green phenotypes is correlated with the cl cl genotype. The y⁺ gene increases the total carotenoids and promotes the formation of red pigments. Giant grana were found in the yellow and green phenotypes, but during ripening these disappeared in the yellow. Unusual dichotomous and concentric grana were observed in the green. Globule-associated carotenoids forming fibrillar crystalloids were present in all color types, although to a lesser degree in the yellow fruit. Membrane-associated carotenoids occurred only in the yellow and green phenotypes.     

Fruit color polymorphism in a bird-dispersed shrub (Rhagodia parabolica) in Australia

Summary We examined the selective basis of fruit color, a trait assumed to affect the attractiveness of fruits to avian dispersal agents, inRhagodia parabolica in Victoria, Australia. The fleshy fruits ofR. parabolica are highly polymorphic in color; individual plants may bear red, white, or yellow fruits, or some combination of these. Red color is produced by betacyanins, yellow by betaxanthins, and white by some other flavonoid compounds. Red is the most common color, white less common, and yellow is relatively rare; red and white is the most common color combination. Fruits of these color-morphs do not differ in maximum size, weight, pulp-seed ratio, water content, or content of major nutrients (sugars, N, and lipids). The most common avian consumer,Zosterops lateralis, foraged at random on the color-morphs, although all fruit-eaters, collectively, slightly favored white fruits. Weekly fruit removal rates were equivalent for all four morphs. Seeds from color-morphs differed in seed germination behavior: seeds from red fruits germinated faster than seeds from yellow fruits, and seeds from white fruits showed the greatest response to passage throughZ. lateralis guts. Lack of strong foraging preferences by birds and the different germination behaviors may contribute to the maintenance of the color polymorphism.     

Identification of two loci involved in phytochrome expression in Nicotiana plumbaginifolia and lethality of the corresponding double mutant

Four Nicotiana plumbaginifolia mutants exhibiting long hypocotyls and chlorotic cotyledons under white light, have been isolated from M2 seeds following mutagenesis with ethyl methane sulphonate. In each of these mutants, this partly etiolated in white light (pew) phenotype is due to a recessive nuclear mutation at a single locus. Complementation analysis indicates that three mutants, dap5, ems28 and ems3-6-34, belong to a single complementation group called pew1, while dap1 defines the pew2 locus. The mutants at pew1 contain normal levels of immunochemically detectable apoprotein of the phytochrome that is relatively abundant in etiolated seedlings, but are deficient in spectrophotometrically detectable phytochrome, whether seedlings are grown in darkness or light. Moreover, biliverdin, a precursor of the phytochrome chromophore, restores light-regulated responses in pew1 mutants and increases their level of photoreversible phytochrome when grown in darkness. These results indicate that the pew1 locus may be involved in chromophore biosynthesis. The mutant at the pew2 locus displays no photoreversible phytochrome in etiolated seedlings, but does contain normal levels of photoreversible phytochrome when grown in the light. Biliverdin had little effect on light-regulated responses in this mutant. In addition, biliverdin did not alter the level of phytochrome in etiolated seedlings. These observations lead us to propose that this mutant could be affected in the phyA gene itself. We have also obtained the homozygous double mutant at the pew1 and pew2 loci. This double mutant is lethal at an early stage of development, consistent with a critical role for phytochrome in early development of higher plants.     

Somatic instability of carotenoid biosynthesis in the tomato ghost mutant and its effect on plastid development

The tomato (Lycopersicon esculentum (L.) Mill.) ghost plant is a mutant of the San Marzano cultivar affected in carotenoid biosynthesis. ghost plants exhibit a variable pattern of pigment biosynthesis during development. Cotyledons are green but true leaves are white. Green sectors, which appear to be clonal in origin, are frequently observed in the white tissue. Because of the lack of photosynthesis ghost plants have a very low viability in soil. We have developed a strategy for propagating ghost plants that employs organ culture to generate variegated green-white plants which, supported by the photosynthetic green areas, develop in soil to almost wild-type size. These plants were used to analyze the pigment content of the different tissues observed during development and plastid ultrastructure. Cotyledons and green leaves contain both colored carotenoids and chlorophyll but only the colorless carotenoid phytoene accumulates in white leaves. the plastids in the white tissue of ghost leaves lack internal membrane structures but normal chloroplasts can be observed in the green areas. The chromoplasts of white fruits are also impaired in their ability to form thylakoid membranes.     

Isolation,characterization, and mapping of the stay green mutant in rice

Leaf color turns yellow during senescence due to the degradation of chlorophylls and photosynthetic proteins. A stay green mutant was isolated from the glutinous japonica rice Hwacheong-wx through N-methyl-N-nitrosourea mutagenesis. Leaves of the mutant remained green, while turning yellow in those of the wild-type rice during senescence. The stay green phenotype was controlled by a single recessive nuclear gene, tentatively symbolized as sgr(t). All the phenotypic characteristics of the mutant were the same as those of the wild-type lines except for the stay green trait. The leaf chlorophyll concentration of the mutant was similar to that of the wild-type before heading, but decreased steeply in the wild-type during grain filling, while very slowly in the mutant. However, no difference in photosynthetic activity was observed between the stay green mutant and the yellowing wild-type leaves, indicating that senescence is proceeding normally in the mutant leaves and that the mutation affects the rate of chlorophyll degradation during the leaf senescence. Using phenotypic and molecular markers, we mapped the sgr(t) locus to the long arm of chromosome 9 between RFLP markers RG662 and C985 at 1.8- and 2.1-cM intervals, respectively. Received: 29 April 2001 / Accepted: 17 July 2001     

ACCLIMATION OF THE PHOTOSYNTHETIC APPARATUS OF PALMARIA PALMATA (RHODOPHYTA) TO LIGHT QUALITIES THAT PREFERENTIALLY EXCITE PHOTOSYSTEM I OR PHOTOSYSTEM II1

Acclimation of the photosynthetic apparatus to light absorbed primarily by phycobilisomes (which transfer energy predominantly to photosystem II) or absorbed by chlorophyll a (mainly present in the antenna of photosystem I) was studied in the macroalga Palmaria palmata L. In addition, the influence of blue and yellow light, exciting chlorophyll a and phycobilisomes, respectively, ivas investigated. All results were compared to a white light control. Complementary chromatic adaptation in terms of an enhanced ratio of phycoerythrin to phycocyanin under green light conditions was observed. Red light (mainly absorbed by chlorophyll a) and green light (mainly absorbed by phycobilisomes) caused an increase of the antenna system, which was not preferentially excited. Yellow and blue light led to intermediate states comparable to each other and white light. Growth was reduced under all light qualities in comparison to white light, especially under conditions preferably exciting phycobilisomes (green light-adapted algae had a 58% lower growth rate compared to white light-adapted algae). Red and blue light-adapted algae showed maximal photosynthetic capacity with white light excitation and significantly lower values with green light excitation. In contrast, green and yellow light-adapted algae exhibited comparable photosynthetic capacities at all excitation wavelengths. Low-temperature fluorescence emission analysis showed an increase of photosystem II emission in red light-adapted algae and a decrease in green light-adapted algae. A small increase of photosystem I emission teas also found in green light-adapted algae, but this was much less than the photosystem II emission increase observed in red light-adapted algae (both compared to phycobilisome emission). Efficiency of energy transfer from phycobilisomes to photosystem II was higher in red than in green light-adapted algae. The opposite was found for the energy transfer efficiency from phycobilisomes to photosystem I. Zeaxanthin content increased in green and blue light-adapted algae compared to red, white, and yellow light-adapted algae. Results are discussed in comparison to published data on unicellular red algae and cyanobacteria.     

Chlorophyll Measurement as a Quantitative Method for the Assessment of Cytokinin-Induced Green Foci Formation in Tissue Culture

Tissue culture systems have long been exploited to study the process of organogenesis. In response to externally applied cytokinins, pluripotent cells proliferate into green calli and subsequently regenerate shoots. Conventionally, the cytokinin-induced greening phenotype has been evaluated by counting numbers of green foci or to present photographic evidence of morphological changes. However, because the structure of calli is disorganized and the development of pigmentation takes place gradually from pale white through yellow to green, adequately defining and counting green foci remains difficult. In this study, we employed chlorophyll measurement as an alternative method to statistically assess the greening phenotype in tissue culture material. We found that N,N-dimethylformamide was the most effective solvent for the extraction of chlorophylls from callus tissue and that bead disruption of the structured tissue improved solvent penetration and the consistency of results. The sensitivity of the method facilitated the quantification of chlorophylls in single-cultured root explants and the use of a spectrophotometer increased the efficiency of measuring multiple samples. Our measurements showed that chlorophyll contents from calli of wild-type and altered cytokinin response mutants (cre1; ahk3, or cytokinin hypersensitive 2 (ckh2)/pickle (pkl) were statistically distinguishable, validating the method. Our proposed procedure represents gains in efficiency and precision and leads to more robust standardization than the conventionally used counting of green foci.

     

Molecular genetics of the y locus in pepper: its relation to capsanthin-capsorubin synthase and to fruit color

Classical genetic studies have determined that the yellow fruit color in pepper is recessive to red in the locus y. We studied the relation of the y locus with the gene coding for capsanthin-capsorubin synthase (CCS) that synthesizes the red carotenoid pigments in the mature fruit. Cosegregation of y and CCS in populations derived from crosses between plants bearing red×white and red×yellow fruits indicated the correspondence of the two genes. We obtained indications for the occurrence of a deletion in the CCS gene in plants containing the recessive y allele. This deletion did not contain the distal 220 bp of the 3′ end of the gene. We used the CCS gene to determine the genotype of peppers with different fruit colors at the y locus. In BC₁ segregants from a red×white cross, the red and peach-fruited progenies had the wild-type allele at the CCS locus, while the orange, yellow and white-fruited progenies had the mutant allele. Screening orange-fruited cultivars with CCS as well as segregation analysis of CCS in an additional red×white cross indicated two possible genotypes of the orange fruit color in this locus. Received: 25 January 1999 / Accepted: 16 August 1999     

Chlorophyll-deficient Lespedeza procumbens

Summary A native plant of Lespedeza procumbens with yellow, chlorophyll-deficient leaves was discovered near Tallahassee, Florida. When selfed this plant yielded only yellow offspring. Normal, green plants of L. procumbens contained about 5.5 times more chlorophyll (dry weight basis) than yellow plants. At high light intensities photosynthetic rates on a chlorophyll basis in yellow plants were 3 to 8 times greater than in green plants. Chloroplasts in yellow plants did not possess high-stacked grana, and the thylakoids displayed unusual cross channels reported previously only in Cassia [7] and Rhodopseudomonas [15]. Chlorophyll deficiency in Lespedeza closely parallels that reported in yellow (Su/su) tobacco [11]. The present paper is the most detailed report on chlorophyll deficiency in a wild plant, all comparable work having been done on cultivars.These studies were partially supported by contract No. AT-(40-1)-2687 from the U.S. Atomic Energy Commission to Professor H. Gaffron.     

Identification of two loci involved in phytochrome expression in Nicotiana plumbaginifolia and lethality of the corresponding double mutant

Four Nicotiana plumbaginifolia mutants exhibiting long hypocotyls and chlorotic cotyledons under white light, have been isolated from M2 seeds following mutagenesis with ethyl methane sulphonate. In each of these mutants, this partly etiolated in white light (pew) phenotype is due to a recessive nuclear mutation at a single locus. Complementation analysis indicates that three mutants, dap5, ems28 and ems3-6-34, belong to a single complementation group called pew1, while dap1 defines the pew2 locus. The mutants at pew1 contain normal levels of immunochemically detectable apoprotein of the phytochrome that is relatively abundant in etiolated seedlings, but are deficient in spectrophotometrically detectable phytochrome, whether seedlings are grown in darkness or light. Moreover, biliverdin, a precursor of the phytochrome chromophore, restores light-regulated responses in pew1 mutants and increases their level of photoreversible phytochrome when grown in darkness. These results indicate that the pew1 locus may be involved in chromophore biosynthesis. The mutant at the pew2 locus displays no photoreversible phytochrome in etiolated seedlings, but does contain normal levels of photoreversible phytochrome when grown in the light. Biliverdin had little effect on light-regulated responses in this mutant. In addition, biliverdin did not alter the level of phytochrome in etiolated seedlings. These observations lead us to propose that this mutant could be affected in the phyA gene itself. We have also obtained the homozygous double mutant at the pew1 and pew2 loci. This double mutant is lethal at an early stage of development, consistent with a critical role for phytochrome in early development of higher plants.     

DYNAMICS OF GENE FLOW IN AN EXPERIMENTAL POPULATION OF CUCUMIS MELO (CUCURBITACEAE)

Gene flow in an experimental garden of Cucumis melo (Cucurbitaceae), the cultivated muskmelon, was measured by placing plants carrying a dominant gene (green cotyledons) in the center of an 18 × 18-m field of recessive plants (yellow cotyledons). At the end of the growing season, all fruits were collected, and seeds planted for analysis of the progeny. The 123 fruits yielded 41,875 seedlings whose genotypes were recorded. Gene flow decreased with increasing distance from the central plot, while the variance increased. The decrease in frequency of the green gene did not follow a leptokurtic pattern over the distance measured, and there was a marked asymmetric and patchy pattern of gene flow in the field. The directionality contributed to a bimodal pattern of gene frequencies among fruits at the edges of the field, some fruits with no green seedlings, some with many. Bumblebee movements were recorded during 4,296 flower visits in an identically arrayed melon field. Bees usually moved to very nearby plants and visited only a few flowers on any one plant. This suggests that pollen carryover from the central patch out was quite high in the test garden. These findings show that local gene flow patterns in plant populations may be complex, and do not always follow regular and symmetrical models. The complex arrangement of progeny genotypes, when acted upon by local selective forces, may contribute to small-scale differences often seen in plant populations.     

Suppression of a key gene involved in chlorophyll biosynthesis by means of virus-inducing gene silencing

The ChlH gene coding the H subunit of magnesium chelatase, an enzyme involved in chlorophyll biosynthesis, was silenced in Nicotiana benthamiana plants by infection with tobacco mosaic virus vectors (pTMV-30b) containing 67, 214 or 549 nt long ChlH inserts. Silencing of the nuclear ChlH gene induced a chimeric phenotype with green and yellow/white leaves associated with alterations of chloroplast shape and ultrastructure. The symptoms became first evident around veins of young leaves, and only later in the mesophyll tissues. The efficiency of gene silencing was not dependent on the insert orientation, but was strongly correlated with the size of the ChlH insert, providing a flexible method to modulate the level of gene suppression. Silencing efficiency seemed to be strongly dependent on endogenous ChlH mRNA level of the target tissue. Silencing of the ChlH gene with the longest fragment of 549 nt also lowered the accumulation of ChlD and chlorophyll synthetase mRNAs, i.e. other genes involved in chlorophyll biosynthesis.     

Genetics and ultrastructure of a cytoplasmically inherited yellow mutant in soybeans

A chimeric plant was observed in the F₂ generation of a cross between a male-sterile line and a plant introduction homozygous for a chromosome interchange in soybeans [Glycine max (L.) Merr.]. F₃ progeny of this plant included one chimera, 36 yellow plants and 16 green plants. The yellow plants, which progressively turn green, were viable and fertile in field, greenhouse and growth-chamber environments. Reciprocal cross-pollinations were made between these yellow plants and four known nuclear yellow mutant plants, between these yellow plants and sibling green plants and between these yellow plants and unrelated green plants. Segregation data from F₁ and F₂ generations indicated cytoplasmic inheritance of the newly discovered yellow phenotype. Pollinations in which reciprocal F₁ hybrid plants were used as male or female parents were made with unrelated green plants. Observations in F₁ and F₂ generations substantiated the hypothesis of cytoplasmic inheritance. No interactions have been observed between this mutant and the various nuclear backgrounds. This is the first report of a cytoplasmically inherited mutant affecting plant color in soybeans. Exchange grafts were made between cytoplasmic yellow plants and sibling green plants and between cytoplasmic yellow plants and unrelated green plants. The phenotype was controlled by the scion, indicating that graft-transmissible agents were not involved. When grown in darkness, cytoplasmic yellow plants and normal green plants accumulated the same amount of protochlorophyllide. Cytoplasmic yellow plants grown in dim light accumulated slightly less chlorophyll than did their green siblings. Electron photomicrographs showed that the prolamellar body (a structure associated with synthesis of protochlorophyllide) and chloroplast ultrastructure were normal in the cytoplasmic yellow mutant. These observations led to the hypothesis that the synchrony involved in deposition of nuclear and cytoplasmic gene products during organelle development is impaired in this cytoplasmic mutant.     

Frugivory and Seed Dispersal by Four Species of Primates in Madagascar's Eastern Rain Forest1

In the unique faunal assemblage of the Malagasy rain forest, lemurs appear to play particularly important roles as seed-dispersing frugivores. A three-month study of feeding ecology and seed dispersal by four species of lemurs in Madagascar's eastern rain forest found that three species, Eulemur rubriventer, Eulemur fulvus, and Varecia variegata were seed dispersers, and the fourth, Propithecus diadema, was a seed predator. In germination trials, seeds passed by lemurs sprouted significantly faster and in greater numbers than those not passed by lemurs. Analysis of fruit morphologies of 69 local plant taxa producing fleshy fruits during the study period found that these fruits fell into two well-defined color categories that correlated significantly with fruit size. Seventy seven percent of fleshy fruits greater than 10 mm in diameter were colored green, brown, tan, purplish, or black, while all fruits less than 10 mm in diameter were colored red, yellow, orange, pink, blue, or white. Three introduced exotic plant species provided exceptions to this pattern, producing fruits which were larger than 10 mm and pink or orange. Fruits chosen by the primates in this study were usually larger than 10 mm in diameter and were in nearly all cases colored green, brown, tan, purplish, red, or some combination of these colors. Morphological traits shared by fruits of multiple plant taxa in the diets of seed-dispersing lemurs suggest possible coevolved relationships between Malagasy rain forest plants and lemurs.     

Characteristics of a virescent cotton mutant

The virescent cotton (Gossypium hirsutum) mutant described here differs from normal cultivated cotton by a single mutation in the nucleus. The mutant exhibits nuclear control of chlorophyll and carotenoid development. Young leaves are distinctly yellow and become green with age. There is no unusual photometabolism of ¹⁴CO₂ or ¹⁴C-acetate in this mutant. It is probable that the nuclear virescent mutation is in a locus concerned with making structural units. The yellow leaves do show a high photosynthetic capacity on a chlorophyll basis. At saturating light intensity the rate of CO₂ fixation is 8 fold higher than the green control leaves. Thus, impaired pigment synthesis which could be lethal is offset by a high photosynthetic capacity in the virescent leaves.