Identification and tissue-specific expression of rutin biosynthetic pathway genes in Capparis spinosa elicited with salicylic acid and methyl jasmonate.

Capparis spinosa is an edible medicinal plant which is considered as an excellent source of rutin. Rutin is a glycoside of the flavonoid quercetin that has been reported to have a beneficial role in controlling various diseases such as hypertension, arteriosclerosis, diabetes, and obesity. In this study, the partial cDNA of four genes involved in the rutin biosynthetic pathway including 4-coumaroyl CoA ligase (4CL), flavonoid 3'-hydroxylase (F3'H), flavonol synthase (FLS) and flavonol-3-O-glucoside L-rhamnosyltransferase (RT) were identified in C.spinosa plants for the first time. The protein sequences of these genes shared high similarity with the same proteins in other plant species. Subsequently, the expression patterns of these genes as well as rutin accumulation in C.spinosa leaves treated with different concentrations of salicylic acid (SA) and methyl jasmonate (MeJA) and also in different tissues of Caper plants treated with 100 mgL-1 SA and 150 µM MeJA were evaluated. The expression of all four genes was clearly up-regulated and rutin contents increased in response to MeJA and SA treatments after 24 h. The highest rutin contents (5.30 mgg-1 DW and 13.27 mgg-1 DW), as well as the highest expression levels of all four genes, were obtained using 100 mgL-1 SA and 150 µM MeJA, respectively. Among the different tissues, the highest rutin content was observed in young leaves treated with 150 µM MeJA, which corresponded to the expression of related genes, especially RT, as a key gene in the rutin biosynthetic pathway. These results suggest that rutin content in various tissues of C. spinosa can be enhanced to a significant extent by MeJA and SA treatments and the gene expression patterns of rutin-biosynthesis-related genes are regulated by these elicitors.

Transcriptome analysis and codominant markers development in caper, a drought tolerant orphan crop with medicinal value.

Caper (Capparis spinosa L.) is a xerophytic shrub cultivated for its flower buds and fruits, used as food and for their medicinal properties. Breeding programs and even proper taxonomic classification of the genus Capparis has been hampered so far by the lack of reliable genetic information and molecular markers. Here, we present the first genomic resource for C. spinosa, generated by transcriptomic approach and de novo assembly. The sequencing effort produced nearly 80 million clean reads assembled into 124,723 unitranscripts. Careful annotation and comparison with public databases revealed homologs to genes with a key role in important metabolic pathways linked to abiotic stress tolerance and bio-compounds production, such purine, thiamine and phenylpropanoid biosynthesis, α-linolenic acid and lipid metabolism. Additionally, a panel of genes involved in stomatal development/distribution and encoding for Stress Associated Proteins (SAPs) was also identified. We also used the transcriptomic data to uncover novel molecular markers for caper. Out of 50 SSRs tested, 14 proved polymorphic and represent the first set of SSR markers for the genus Capparis. This transcriptome will be an important contribution to future studies and breeding programs for this orphan crop, aiding to the development of improved varieties to sustain agriculture in arid conditions.

Genetic relationships among subspecies of Capparis spinosa L. from Tunisia by using ISSR markers.

In Tunisia, Capparis spinosa L. is widely distributed in different geographic areas. Although it has been extensively studied morphologically, the phylogenetic relationships by using molecular markers among Capparis taxa are still unclear. The Inter-Simple Sequence Repeats (ISSR) molecular markers were used to assess the genetic relationship of this species cultivated in the North and the South of Tunisia. Fifteen ISSR primers were analyzed to characterize the genetic profiles of 67 different genotypes. The ISSR markers produced 108 bands ranging from 4 (ISSR8) to 11 (IAM12 and ISSR16) with an average of 7.2. The observed heterozygosity ranged from 0.43 to 0.95 for ISSR7 and IAM12, respectively. The Polymorphic Information Content (PIC) ranged from 0.48 at the UBC808 to 0.85 at IAM12 and eight loci could be classified as useful for genetic mapping (PIC > 0.7). The genetic diversity within a population was high and varied according to the subspecies and bioclimatic zones. Both UPGMA (Unweighted Pair Group Method with Arithmetic mean) and PCoA (Principal Coordinate Analysis) analyses showed that populations from each subspecies grouped together. However, the structure analysis generated more groups than the PCoA plot and UPGMA, which revealed the mixed allelic of the species Capparis spinosa in Tunisia.

Anti-inflammatory potential of Capparis spinosa L. in vivo in mice through inhibition of cell infiltration and cytokine gene expression.

BACKGROUND: Several chronic inflammatory diseases are characterized by inappropriate CD4+ T cell response. In the present study, we assessed the ability of Capparis spinosa L. (CS) preparation to orientate, in vivo, the immune response mediated by CD4+ T cells towards an anti-inflammatory response. METHODS: The in vivo study was carried out by using the contact hypersensitivity (CHS) model in Swiss mice. Then we performed a histological analysis followed by molecular study by using real time RT-PCR. We also realized a phytochemical screening and a liquid-liquid separation of CS preparation. RESULTS: Our study allowed us to detect a significantly reduced edema in mice treated with CS preparations relative to control. CS effect was dose dependent, statistically similar to that observed with indomethacin, independent of the plant genotype and of the period of treatment. Furthermore, our histology studies revealed that CS induced a significant decrease in immune cell infiltration, in vasodilatation and in dermis thickness in the inflammatory site. Interestingly, we showed that CS operated by inhibiting cytokine gene expression including IFNγ, IL-17 and IL-4. Besides, phytochemical screening of CS extract showed the presence of several chemical families such as saponins, flavonoids and alkaloids. One (hexane fraction) out of the three distinct prepared fractions, exhibited an anti-inflammatory effect similar to that of the raw preparation, and would likely contain the bioactive(s) molecule(s). CONCLUSIONS: Altogether, our data indicate that CS regulates inflammation induced in vivo in mice and thus could be a source of anti-inflammatory molecules, which could be used in some T lymphocyte-dependent inflammatory diseases.

Genetic diversity and population structure of six species of Capparis in Tunisia using AFLP markers.

In order to study the genetic diversity, the phylogeographic pattern and hybridization between six Tunisian Capparis species, 213 accessions of Caper were genotyped with three primer combinations of amplified fragment length polymorphism (AFLP) markers. Out of 750 fragments generated, 636 were polymorphic and 407 of them were restricted to a single species. STRUCTURE and PCoA analyses clearly separated morphologically different populations into six distinct genetic ones. The UPGMA analysis grouped the species into three main clusters: G1 grouped C. spinosa subsp. spinosa var. spinosa and C. sicula subsp. sicula; G2 grouped C. ovata subsp. ovata and C. orientalis and G3 clustered C. zoharyi and C. aegyptia. Populations from G1, G2 and G3 were mainly distributed in arid, subhumid, and semi-arid bioclimates, respectively. Additional genetic studies on Capparis could help to identify genes underlying speciation events and local adaptation to geographic areas leading to the development of breeding programs.

The Multifarious PGPR Serratia marcescens CDP-13 Augments Induced Systemic Resistance and Enhanced Salinity Tolerance of Wheat (Triticum aestivum L.).

The present study demonstrates the plant growth promoting (PGP) potential of a bacterial isolate CDP-13 isolated from 'Capparis decidua' plant, and its ability to protect plants from the deleterious effect of biotic and abiotic stressors. Based on 16S rRNA gene sequence analysis, the isolate was identified as Serratia marcescens. Among the PGP traits, the isolate was found to be positive for ACC deaminase activity, phosphate solubilization, production of siderophore, indole acetic acid production, nitrogen fixation, and ammonia production. CDP-13 showed growth at an increased salt (NaCl) concentration of up to 6%, indicating its potential to survive and associate with plants growing in saline soil. The inoculation of S. marcescens enhanced the growth of wheat plant under salinity stress (150-200 mM). It significantly reduced inhibition of plant growth (15 to 85%) caused by salt stressors. Application of CDP-13 also modulated concentration (20 to 75%) of different osmoprotectants (proline, malondialdehyde, total soluble sugar, total protein content, and indole acetic acid) in plants suggesting its role in enabling plants to tolerate salt stressors. In addition, bacterial inoculation also reduced the disease severity caused by fungal infection, which illustrated its ability to confer induced systemic resistance (ISR) in host plants. Treatment of wheat plants with the test organism caused alteration in anti-oxidative enzymes activities (Superoxide dismutase, Catalase, and Peroxidase) under various salinity levels, and therefore minimizes the salinity-induced oxidative damages to the plants. Colonization efficiency of strain CDP-13 was confirmed by CFU count, epi-fluorescence microscopy, and ERIC-PCR-based DNA fingerprinting approach. Hence, the study indicates that bacterium CDP-13 enhances plant growth, and has potential for the amelioration of salinity stress in wheat plants. Likewise, the results also provide insights into biotechnological approaches to using PGPR as an alternative to chemicals and pesticides.

Genetic diversity analysis of Capparis spinosa L. populations by using ISSR markers.

Capparis spinosa L. is an important medicinal species in the Xinjiang Province of China. Ten natural populations of C. spinosa from 3 locations in North, Central, and South Xinjiang were studied using morphological trait inter simple sequence repeat (ISSR) molecular markers to assess the genetic diversity and population structure. In this study, the 10 ISSR primers produced 313 amplified DNA fragments, with 52% of fragments being polymorphic. Unweighted pair-group method with arithmetic average (UPGMA) cluster analysis indicated that 10 C. spinosa populations were clustered into 3 geographically distinct groups. The Nei gene of C. spinosa populations in different regions had Diversity and Shannon's information index ranges of 0.1312-0.2001 and 0.1004-0.1875, respectively. The 362 markers were used to construct the dendrogram based on the UPGMA cluster analysis. The dendrogram indicated that 10 populations of C. spinosa were clustered into 3 geographically distinct groups. The results showed these genotypes have high genetic diversity, and can be used for an alternative breeding program.

Physicochemical properties of caper species seed oils collected from two different harvest years.

BACKGROUND: In this study the physicochemical properties, fatty acid composition and sterol and tocopheol contents of caper species seed oils were determined. RESULTS: Brightness (L*) values of oils obtained from caper seeds ranged between 36.76 and 53.48. Viscosity values of oils of Capparis spinosa species were between 41.1 and 48.6 mPa. While oleic acid levels were high in C. spinosa species oils, linoleic acid levels were high in Capparis ovata species oils. Sterol contents of crude oils were between 3140 and 3272 mg kg(-1) (mean 3220 mg kg(-1)) for C. spinosa species and between 3275 and 3312 mg kg(-1) (mean 3298 mg kg(-1)) for C. ovata species. α-Tocopherol contents of C. spinosa and C. ovata oils averaged 3.87 and 2.63 mg per 100 g respectively. Radical-scavenging activities of crude oils averaged 29.78% (C. spinosa species) and 26.09% (C. ovata species). Total phenolic concentrations in crude oils averaged 0.642 mg g(-1) for C. ovata species (P < 0.01). CONCLUSION: Caper seeds are a natural source of vegetable oils that are beneficial in terms of health, oil stability and resistance to oxidation.

Combining molecular-marker and chemical analysis of Capparis decidua (Capparaceae) in the Thar Desert of Western Rajasthan (india).

The Thar Desert, a very inhospitable place, accommodates only plant species that survive acute drought, unpredictable precipitation, and those can grow in the limited moisture of sandy soils. Capparis decidua is among one of the few plants able to grow well under these conditions. This species is highly exploited and has been naturally taken, as local people use it for various purposes like food, timber and fuel, although, no management or conservation efforts have been established. The present study was conducted in this arid area of Western Rajasthan (India) with the aim to obtain preliminary molecular information about this group of plants. We evaluated diversity among 46 samples of C. decidua using chemical parameters and random amplified polymorphic DNA (RAPD) markers. Fourteen chemical parameters and eight minerals (total 22 variables) of this species fruits were estimated. A total of 14 RAPD primers produced 235 band positions, of which 81.27% were polymorphic. Jaccard's similarity coefficients for RAPD primers ranged from 0.34 to 0.86 with a mean genetic similarity of 0.50. As per observed coefficient of variation, NDF (Neutral Detergent Fiber) content was found to be the most variable trait followed by starch and soluble carbohydrate. The Manhattan dissimilarity coefficient values for chemical parameters ranged between 0.02-0.31 with an average of 0.092. The present study revealed a very low correlation (0.01) between chemical parameters and RAPD-based matrices. The low correlation between chemical- and RAPD-based matrices indicated that the two methods were different and highly variable. The chemical-based diversity will assist in selection of nutritionally rich samples for medicinal purpose, while genetic diversity to face natural challenges and find sustainable ways to promote conservation for future use.

Combining molecular-marker and chemical analysis of Capparis deciduas (Capparaceae) in the Thar Desert of Western Rajasthan (India)

The Thar Desert, a very inhospitable place, accommodates only plant species that survive acute drought, unpredictable precipitation, and those can grow in the limited moisture of sandy soils. Capparis decidua is among one of the few plants able to grow well under these conditions. This species is highly exploited and has been naturally taken, as local people use it for various purposes like food, timber and fuel, although, no management or conservation efforts have been established. The present study was conducted in this arid area of Western Rajasthan (India) with the aim to obtain preliminary molecular information about this group of plants. We evaluated diversity among 46 samples of C. decidua using chemical parameters and random amplified polymorphic DNA (RAPD) markers. Fourteen chemical parameters and eight minerals (total 22 variables) of this species fruits were estimated. A total of 14 RAPD primers produced 235 band positions, of which 81.27% were polymorphic. Jaccard s similarity coefficients for RAPD primers ranged from 0.34 to 0.86 with a mean genetic similarity of 0.50. As per observed coefficient of variation, NDF (Neutral Detergent Fiber) content was found to be the most variable trait followed by starch and soluble carbohydrate. The Manhattan dissimilarity coefficient values for chemical parameters ranged between 0.02-0.31 with an average of 0.092. The present study revealed a very low correlation (0.01) between chemical parameters and RAPD-based matrices. The low correlation between chemical- and RAPD-based matrices indicated that the two methods were different and highly variable. The chemical-based diversity will assist in selection of nutritionally rich samples for medicinal purpose, while genetic diversity to face natural challenges and find sustainable ways to promote conservation for future use.

El desierto de Thar, un lugar muy inhóspito, alberga sólo a las especies de plantas capaces de resistir a condiciones de sequía extrema, a las precipitaciones impredecibles, y a las plantas que pueden crecer en la humedad limitada de los suelos arenosos. Capparis decidua se encuentra entre una las pocas plantas capaces de crecer bien en estas condiciones. Esta especie es altamente explotada y se ha tomado de forma natural, así los habitantes locales las han usado para varios propósitos, como alimento, madera y combustible, aunque sin ningún programa de manejo o esfuerzo por conservación. El presente estudio se llevó a cabo en esta zona árida del oeste de Rajastán (India) con el objetivo de obtener información molecular preliminar sobre este grupo de plantas. Se evaluó la diversidad entre 46 muestras de C. decidua usando parámetros químicos y marcadores de ADN polimórfico amplificado al azar (RAPD por sus siglas en inglés). Catorce parámetros químicos y ocho minerales (22 variables en total) de los frutos de esta especie fueron estimados. Un total de 14 cebadores para RAPD produjeron 235 posiciones de bandas, de las cuales 81.27% fueron polimórficas. El coeficiente de similitud de Jaccard para los cebadores del RAPD varió entre 0.34 y 0.86 con un promedio de similitud genética de 0.50. De acuerdo con el coeficiente de variación observado, se encontró que el contenido de NDF fue el rasgo más variable, seguido por el almidón y los carbohidratos solubles. Los valores del coeficiente de disimilitud de Manhattan para los parámetros químicos osciló entre 0.02-0.31 con un promedio de 0.092. El presente estudio reveló una correlación muy baja (0.01) entre los parámetros químicos y las matrices basadas en RAPD. La baja correlación entre las matrices químicas y la basada en RAPD indicó que los dos métodos fueron diferentes y altamente variables. El estudio de la diversidad basada en su química ayudará en la selección de muestras nutricionalmente ricas para fines medicinales, mientras que la diversidad genética ayudará a enfrentar los desafíos naturales y encontrar formas sostenibles para promover la conservación de esta plana para uso futuro.

Embryo and anther regulation of the mabinlin II sweet protein gene in Capparis masaikai Lévl.

Mabinlin II is one of the major sweet proteins stored in the seeds of Capparis masaikai Lévl. Its promoter region (779 bp) located 5' upstream of the mabinlin II gene has been isolated and named as MBL-779 (GenBank accession number, EU014073). This promoter contains two typical TATA box regions and a series of motifs related to seed-specific promoters, such as ACGT motifs, RY motif, napin motif, and G box. The MBL-779 promoter drove GUS gene to transiently express in the embryos of bean, maize, and rice seeds or to constantly express in the embryos and anthers of the transgenic Arabidopsis. The MBL-779 promoter regulated gene expression from approximately the 12th day and peaked on approximately the 16th day after flowering in Arabidopsis. The -300-bp promoter region is a minimal sequence required to functionally regulate gene expression. The CAATs at -325 to -322 bp and -419 to -416 bp and the region at -485 to -770 bp play a role in the quantitative regulation of gene expression. The RY motif, CATGAC, at -117 to -112 bp and the ACGT within the G box (CACGTG) at -126 to -123 bp positively regulate gene expression.