Flavonoid-Deficient Mutants in Grass Pea (Lathyrus sativus L.): Genetic Control, Linkage Relationships, and Mapping with Aconitase and S-Nitrosoglutathione Reductase Isozyme Loci
19 Apr 2012-The Scientific World Journal (Hindawi Publishing Corporation)-Vol. 2012, pp 345983-345983
TL;DR: Two flavonoid-deficient mutants were isolated in EMS-mutagenized M2 progeny of grass pea, revealing monogenic recessive inheritance of the trait and mapping Aco 1 and fld 1 loci on extra chromosome of trisomic-I and Aco 2, fld 2, and Gsnor 2 in linked associations.
Abstract: Two flavonoid-deficient mutants, designated as fldL-1 and fldL-2, were isolated in EMS-mutagenized (0.15%, 10 h) M2 progeny of grass pea (Lathyrus sativus L.). Both the mutants contained total leaf flavonoid content only 20% of their mother varieties. Genetic analysis revealed monogenic recessive inheritance of the trait, controlled by two different nonallelic loci. The two mutants differed significantly in banding patterns of leaf aconitase (ACO) and S-nitrosoglutathione reductase (GSNOR) isozymes, possessing unique bands in Aco 1, Aco 2, and Gsnor 2 loci. Isozyme loci inherited monogenically showing codominant expression in F2 (1 : 2 : 1) and backcross (1 : 1) segregations. Linkage studies and primary trisomic analysis mapped Aco 1 and fld 1 loci on extra chromosome of trisomic-I and Aco 2, fld 2, and Gsnor 2 on extra chromosome of trisomic-IV in linked associations.
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TL;DR: The role of NO in the process of amelioration has ultimately been manifested by significant reduction of membrane damage and improvement of growth performance in plants grown on As + SNP media.
Abstract: The adverse effects of arsenic (As) toxicity on seedling growth, root and shoot anatomy, chlorophyll and carotenoid contents, root oxidizability (RO), antioxidant enzyme activities, H2O2 content, lipid peroxidation and electrolyte leakage (EL%) in common bean (Phaseolus vulgaris L.) were investigated. The role of exogenous nitric oxide (NO) in amelioration of As-induced inhibitory effect was also evaluated using sodium nitroprusside (100 μM SNP) as NO donor and 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (200 μM PTIO) as NO scavenger in different combinations with 50 μM As. As-induced growth inhibition was associated with marked anomalies in anatomical features, reduction in pigment composition, increased RO and severe perturbations in antioxidant enzyme activities. While activity of superoxide dismutase and catalase increased, levels of ascorbate peroxidase, dehydroascorbate reductase and glutathione reductase decreased significantly and guaiacol peroxidase remained normal. The over-accumulation of H2O2 content along with high level of lipid peroxidation and electrolyte leakage indicates As-induced oxidative damage in P. vulgaris seedlings with more pronounced effect on the roots than the shoots. Exogenous addition of NO significantly reversed the As-induced oxidative stress, maintaining H2O2 in a certain level through balanced alterations of antioxidant enzyme activities. The role of NO in the process of amelioration has ultimately been manifested by significant reduction of membrane damage and improvement of growth performance in plants grown on As + SNP media. Onset of oxidative stress was more severe after addition of PTIO, which confirms the protective role of NO against As-induced oxidative damage in P. vulgaris seedlings.
146 citations
Cites background from "Flavonoid-Deficient Mutants in Gras..."
...Decreased activity of APX might be either due to low availability of reduced ascorbate, its co-factor, or inhibition of its isoform/s by excess H2O2 or both (Hiner et al. 2000; Talukdar 2012b)....
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...The gaseous free radical nitric oxide (NO) is a widespread intracellular and intercellular messenger with a broad spectrum of regulatory roles in plant physiological processes (Wendehenne et al. 2001; Neill et al. 2002; Talukdar 2012a)....
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...Recently, this group of plants has been identified as one of the dominant legume taxa, which is being used in Eastern Himalayas for different types of ethnic medicinal and edible purposes (Talukdar and Talukdar 2012)....
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...POX is able to catalyze lignin synthesis and is induced in higher plants exposed to toxic metals or other stresses (Prasad 1996; Probst et al. 2009; Talukdar 2012b)....
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...Significant increase in MDA content and loss in photosynthetic apparatus have been recognized as the marks of oxidative stress (Mascher et al. 2002; Li et al. 2006; Talukdar 2011b, 2012a) and may be one of the prime reasons for As-induced growth inhibition in the present material....
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TL;DR: Ca priming in the media significantly reduced the Cd accumulation and considerably alleviated the adverse impact of Cd treatment by modulating the antioxidant enzyme activity, which enhanced fresh mass of plant parts as the sign of Ca-mediated normal growth in Cd-treated lentil seedlings.
Abstract: The effect of calcium (Ca) on lentil (Lens culinaris Medic.) seedlings exposed to cadmium (Cd) stress was studied by investigating plant growth and antioxidant enzyme activities. Plants were grown for 14 days in full-strength Hoagland nutrient media supplemented with Cd concentrations of 0, 10, 20, and 40 μM, and on corresponding medium supplied with 5 mM Ca(NO3)2 prior to Cd addition. Increasing Cd led to accumulation of metal and reduced the fresh weight of the shoots more strongly than that of the roots. Cd concentrations of 20 and 40 μM were selected to study its toxic effect on seedlings. Activities of superoxide dismutase, ascorbate peroxidase, catalase, dehydroascorbate reductase, and glutathione reductase decreased at much higher magnitude in the shoots than those observed in the roots under Cd exposure. Failure of antioxidant defense in scavenging of reactive oxygen species was evidenced by abnormal rise in H2O2, resulting in enhancement of lipid peroxidation and membrane electrolyte leakage as the marks of Cd-induced oxidative stress in lentil seedlings. Ca priming in the media significantly reduced the Cd accumulation and considerably alleviated the adverse impact of Cd treatment by modulating the antioxidant enzyme activity. Mitigation of Cd-induced stress by Ca application was strongly suggested by declining levels of H2O2 and consequent lowering of oxidative damage of membrane. Consequently, this enhanced fresh mass of plant parts as the sign of Ca-mediated normal growth in Cd-treated lentil seedlings.
90 citations
Cites background from "Flavonoid-Deficient Mutants in Gras..."
...However, Cd can raise the basal ROS level either by inactivation or downregulation of ROS-scavenging antioxidant enzymes and metabolites (Clemens 2006; Sanita di Toppi and Gabrielli 1999; Talukdar 2012c)....
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...The response of these compounds, however, largely depends on both external and internal stimuli and also genetic backgrounds of the plants under stress (Talukdar 2011a, 2012a), and their modulation ultimately determines plant growth and development (De Pinto and De Gara 2004)....
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...…and Bhattacharyya 2012; Roy et al. 2010) and its toxicity on growth of many leguminous crops (Anjum et al. 2011; Rodríguez-Serrano et al. 2009; Sandalio et al. 2001; Talukdar 2012c) the effect of this heavy metal on response of antioxidant defense and the process of amelioration needs urgent study....
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...…in favor of a reduced state, which plays a pivotal role in the maintenance of root growth under stress conditions as has been reported earlier in Arabidopsis (Vernoux et al. 2000) and in the leguminous plant, Lathyrus sativus L. (Talukdar 2011a, 2012b, c) and Phaseolus vulgaris L. (Talukdar 2012d)....
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TL;DR: This review describes progress achieved in Lathyrus genetic improvement and explores potential opportunities and urgent research needs for this somewhat neglected crop.
Abstract: Grass pea offers an attractive choice for sustainable food production, owing to its intrinsic properties including limited water requirement and drought tolerance. However, low productivity and the presence of a neurotoxin (ODAP) have posed major obstacles to its genetic improvement. Also, biotechnological investments remain limited and the genome is complex and not well understood. Strategies that allow identification of genotypes with reduced ODAP content, coupling of low ODAP content with enhanced yield, and effective seed detoxification methods merit immediate attention. Breeder-friendly genomic tools are being increasingly made available to improve the efficiency of breeding protocols. To this end, the application of next-generation sequencing has provided a means of leveraging the repertoire of genomic resources for this somewhat neglected crop. In this review, we describe progress achieved in Lathyrus genetic improvement. We also explore potential opportunities in Lathyrus research and identify urgent research needs.
67 citations
TL;DR: This article examines the prospects of genomics assisted integrated breeding to enhance and stabilize crop yields and outlines the recent progress made in genomics of these lesser explored pulse crops.
Abstract: Pulses are multipurpose crops for providing income, employment and food security in the underprivileged regions, notably the FAO-defined low-income food-deficit countries. Owing to their intrinsic ability to endure environmental adversities and the least input/management requirements, these crops remain central to subsistence farming. Given their pivotal role in rain-fed agriculture, substantial research has been invested to boost the productivity of these pulse crops. To this end, genomic tools and technologies have appeared as the compelling supplement to the conventional breeding. However, the progress in minor pulse crops including dry beans (Vigna spp.), lupins, lablab, lathyrus and vetches has remained unsatisfactory, hence these crops are often labeled as low profile or lesser researched. Nevertheless, recent scientific and technological breakthroughs particularly the next generation sequencing (NGS) are radically transforming the scenario of genomics and molecular breeding in these minor crops. NGS techniques have allowed de novo assembly of whole genomes in these orphan crops. Moreover, the availability of a reference genome sequence would promote re-sequencing of diverse genotypes to unlock allelic diversity at a genome-wide scale. In parallel, NGS has offered high-resolution genetic maps or more precisely, a robust genetic framework to implement whole-genome strategies for crop improvement. As has already been demonstrated in lupin, sequencing-based genotyping of the representative sample provided access to a number of functionally-relevant markers that could be deployed straight away in crop breeding programs. This article attempts to outline the recent progress made in genomics of these lesser explored pulse crops, and examines the prospects of genomics assisted integrated breeding to enhance and stabilize crop yields.
33 citations
TL;DR: Impact of water stress was more severe on lentil compared with grass pea, and modulation of growth traits signified necessity of a detailed strategy in breeding of food legumes under water stress.
Abstract: Background: Both lentil (Lens culinaris Medik.) and grass pea (Lathyrus sativus L.) in the family Fabaceae are two important cool-season food legumes, often experiencing water stress conditions during growth and maturity. Objective: The present study was undertaken to ascertain the response of these two crops under different water stress regimes. Materials and Methods: Different morpho-physiological and biochemical parameters were studied in a pot experiment under controlled environmental conditions. Along with control (proper irrigation, 0 stress), three sets of plants were subjected to mild (6 d), moderate (13 d) and severe (20 d) water stress by withholding irrigation at the appropriate time. Results: Compared with control, plant growth traits and seed yield components reduced significantly in both crops with increasing period of water stress, resulting in lowering of dry mass with more severe effect on lentil compared with grass pea. Foliar Relative Water Content (RWC) (%), K + /Na + ratio, chlorophyll (chl) a, chl a/b ratio, stomatal conductance and net photosynthetic rate declined considerably in both crops under water stress. Leaf-free proline level increased significantly in both crops, but it decreased markedly in nodules of lentil and remained unchanged in grass pea. Nodulation was also affected due to water stress. The impairment in growth traits and physio-biochemical parameters under water stress was manifested in reduction of drought tolerance efficiency of both crops. Conclusion: Impact of water stress was more severe on lentil compared with grass pea, and modulation of growth traits signified necessity of a detailed strategy in breeding of food legumes under water stress.
28 citations
Cites background from "Flavonoid-Deficient Mutants in Gras..."
...[12] In lentil, cadmium-induced oxidative stress and its amelioration by calcium has been studied,[15] while two flavonoid-deficient mutants, one glutathione-deficient and one overproducing mutant, and one ascorbate-deficient semi-dwarf mutant have recently been characterized in grass pea.[16-19] These valuable biochemical mutations are now giving vital clues about the intrinsic tolerant mechanism of legume crops and are now being mapped using the arrays of cytological tester stocks such as aneuploids, translocations, and polyploids....
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References
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TL;DR: The genetic map is a tool to quantify the distance between genes on a chromosome, based on the observed frequency of crossovers during cell division, which is used to estimate the total distance between chromosomes.
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5,845 citations
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4,046 citations
"Flavonoid-Deficient Mutants in Gras..." refers background in this paper
...The modification of flower colour was ascribed to the deficiency of anthocyanin biosynthesis and is of considerable assistance in plant breeding [45]....
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TL;DR: The characterization of genetically defined mutations has enabled the order of many reactions in anthocyanin synthesis and their modification to be elucidated, and the more recent developments in gene isolation and characterization are concentrated.
Abstract: Flavonoids represent a large class of secondary plant metabolites, of which anthocyanins are the most conspicuous class, dueto the wide range of colors resulting from their synthesis. Anthocyanins are important to many diverse functions within plants. Synthesis of anthocyanins in petals is undoubtedly intended to attract pollinators, whereas anthocyanin synthesis in seeds and fruits may aid in seed dispersal. Anthocyanins and other flavonoids can also be important as feeding deterrents and as protection against damage from UV irradiation. The existence of such a diverse range of functions and types of anthocyanins raises questions about how these compounds are synthesized and how their synthesis is regulated. The study of the genetics of anthocyanin synthesis began last century with Mendel’s work on flower color in peas. Since that time, there have been periods of intensive study into the genetics and biochemistry of pigment production in a number of different species. In the early studies, genetic loci were correlated with easily observable color changes. After the structures of anthocyanins and other flavonoids were determined, it was possible to correlate single genes with particular structural alterations of anthocyanins or with the presence or absence of particular flavonoids. Mutations in anthocyanin genes have been studied for many years because they are easily identified and because they generally have no deleterious effect on plant growth and development. In most cases, mutations affecting different steps of the anthocyanin biosynthesis pathway were isolated and characterized well before their function was identified or the corresponding gene was isolated. More recently, many genes involved in the biosynthesis of anthocyanin pigments have been isolated and characterized using recombinant DNA technologies. Three species have been particularly important for elucidating the anthocyanin biosynthetic pathway and for isolating genes controlling the biosynthesis of flavonoids: maize (Zea mays), snapdragon (Anfirrhinum majus), and petunia (Wtunia hybrida). Petunia has more recently become the organism of choice for isolating flavonoid biosynthetic genes and studying their interactions and regulation. At least 35 genes are known to affect flower color in petuniawiering and de Vlaming, 1984). Because this field of research has been reviewed fairly extensively in the past (Dooner et al., 1991; van Tunen and MOI, 1991; Gerats and Martin, 1992), in this review we concentrate on the more recent developments in gene isolation and characterization. A review of the genetics of flavonoid biosynthesis in other species was recently covered by Forkmann (1993). The characterization of genetically defined mutations has enabled the order of many reactions in anthocyanin synthesis and their modification to be elucidated. Some reactions have been postulated only on the basis of genetic studies and have not yet been demonstrated in vitro. Chemico-genetic studies have been very important in determining the enzymatic steps involved in anthocyanin biosynthesis and modification. The generation of transposon-tagged mutations and the subsequent cloning of the transposons provided a relatively straightforward means of isolating many genes from maize (Wienand et al., 1990) and snapdragon (Martin et al., 1991). However, a number of genes in the pathway have not been amenable to transposon tagging. Anthocyanin biosynthetic genes have been isolated using a range of methodologies, including protein purification, transposon tagging, differential screening, and polymerase chain reaction (PCR) amplification. Functions of isolated anthocyanin genes can be confirmed by restriction fragment length polymorphism (RFLP) mapping, complementation, or expression in heterologous systems. Reverse genetics has also been used recently to identify gene function; this requires a welldefined pathway to correlate phenotype with gene function. Once a gene has been isolated from one species, it is usually a straightforward task to isolate the homologous gene from other species by using the original clone as a molecular probe.
1,405 citations
"Flavonoid-Deficient Mutants in Gras..." refers background in this paper
...The pigmented flavonoid metabolites have been used as phenotypic markers in many model plant species [3, 4] and have proven to be an excellent tool to study the genetic, molecular, and biochemical processes [4, 5]....
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TL;DR: The analysis of pigmentation continues to provide insights into new areas, such as the channeling and intracellular transport of metabolites, regulation of gene expression and RNA interference.
Abstract: For more than a century, the biosynthesis of flavonoid pigments has been a favorite of scientists to study a wide variety of biological processes, such as inheritance and transposition, and has become one of the best-studied pathways in nature. The analysis of pigmentation continues to provide insights into new areas, such as the channeling and intracellular transport of metabolites, regulation of gene expression and RNA interference. Moreover, because pigmentation is studied in a variety of species, it provides unique molecular insights into the evolution of biochemical pathways and regulatory networks.
1,359 citations
"Flavonoid-Deficient Mutants in Gras..." refers background in this paper
...The pigmented flavonoid metabolites have been used as phenotypic markers in many model plant species [3, 4] and have proven to be an excellent tool to study the genetic, molecular, and biochemical processes [4, 5]....
[...]
01 Jan 1995
TL;DR: In this paper, the authors focus on the more recent developments in gene isolation and characterization of anthocyanin biosynthetic genes and study their interactions and regulation in different species of maize, snapdragon, and petunia.
Abstract: Flavonoids represent a large class of secondary plant metabolites, of which anthocyanins are the most conspicuous class, dueto the wide range of colors resulting from their synthesis. Anthocyanins are important to many diverse functions within plants. Synthesis of anthocyanins in petals is undoubtedly intended to attract pollinators, whereas anthocyanin synthesis in seeds and fruits may aid in seed dispersal. Anthocyanins and other flavonoids can also be important as feeding deterrents and as protection against damage from UV irradiation. The existence of such a diverse range of functions and types of anthocyanins raises questions about how these compounds are synthesized and how their synthesis is regulated. The study of the genetics of anthocyanin synthesis began last century with Mendel’s work on flower color in peas. Since that time, there have been periods of intensive study into the genetics and biochemistry of pigment production in a number of different species. In the early studies, genetic loci were correlated with easily observable color changes. After the structures of anthocyanins and other flavonoids were determined, it was possible to correlate single genes with particular structural alterations of anthocyanins or with the presence or absence of particular flavonoids. Mutations in anthocyanin genes have been studied for many years because they are easily identified and because they generally have no deleterious effect on plant growth and development. In most cases, mutations affecting different steps of the anthocyanin biosynthesis pathway were isolated and characterized well before their function was identified or the corresponding gene was isolated. More recently, many genes involved in the biosynthesis of anthocyanin pigments have been isolated and characterized using recombinant DNA technologies. Three species have been particularly important for elucidating the anthocyanin biosynthetic pathway and for isolating genes controlling the biosynthesis of flavonoids: maize (Zea mays), snapdragon (Anfirrhinum majus), and petunia (Wtunia hybrida). Petunia has more recently become the organism of choice for isolating flavonoid biosynthetic genes and studying their interactions and regulation. At least 35 genes are known to affect flower color in petuniawiering and de Vlaming, 1984). Because this field of research has been reviewed fairly extensively in the past (Dooner et al., 1991; van Tunen and MOI, 1991; Gerats and Martin, 1992), in this review we concentrate on the more recent developments in gene isolation and characterization. A review of the genetics of flavonoid biosynthesis in other species was recently covered by Forkmann (1993). The characterization of genetically defined mutations has enabled the order of many reactions in anthocyanin synthesis and their modification to be elucidated. Some reactions have been postulated only on the basis of genetic studies and have not yet been demonstrated in vitro. Chemico-genetic studies have been very important in determining the enzymatic steps involved in anthocyanin biosynthesis and modification. The generation of transposon-tagged mutations and the subsequent cloning of the transposons provided a relatively straightforward means of isolating many genes from maize (Wienand et al., 1990) and snapdragon (Martin et al., 1991). However, a number of genes in the pathway have not been amenable to transposon tagging. Anthocyanin biosynthetic genes have been isolated using a range of methodologies, including protein purification, transposon tagging, differential screening, and polymerase chain reaction (PCR) amplification. Functions of isolated anthocyanin genes can be confirmed by restriction fragment length polymorphism (RFLP) mapping, complementation, or expression in heterologous systems. Reverse genetics has also been used recently to identify gene function; this requires a welldefined pathway to correlate phenotype with gene function. Once a gene has been isolated from one species, it is usually a straightforward task to isolate the homologous gene from other species by using the original clone as a molecular probe.
1,345 citations