Marginal quality water arbitrated essential oil contents in metal hoarded flower petals of scented roses.

Climate change and the consequent alteration in agricultural circumstances enhance the susceptibility of fresh water use particularly in water-scarce regions. Marginal quality water reuse is a common alternative practice but possible perils of metal accretion in plant parts are mostly ignored. The present research aimed to probe the impact of treated wastewater (TWW) and untreated wastewater (UTWW) on metal accumulation in flower petals and their influence on essential oil contents of fragrant Rosa species (R. Gruss-an-teplitz, R. bourboniana, R. centifolia, R. damascena) in a peri-urban area of Faisalabad, Pakistan during January, 2017 to December, 2018. The mineral and chemical contents in canal water (CW) and TWW were less than recommended levels of national environmental quality standards (NEQS) for wastewater of Pakistan. The experimentally UTWW possessed higher electrical conductivity (EC), biological and chemical oxygen demand (BOD and COD), and some metals (Pb, Co, Cr) that were above the permissible levels. The experimental data revealed that except Cr other metals contents in the flower petals were less than the WHO recommended limits (for medicinal plants) under experimental irrigation regimes. Rosa centifolia and R. damascena possessed higher metal i.e. Zn, Cu, Pb, Cr, Co contents while Fe and Ni contents were higher in R. Gruss-an-Teplitz and R. bourboniana respectively. There were twelve constituents which were detected in essential oil by gas chromatography. Major constituents were phenyl ethyl alcohol, citronellol, geranyl acetate, γ- undelactone, methyl eugenol, and limonene whose share was 48.17%, 41.11%, 8.46%, 4.82%, 4.44%, and 4.15% respectively whereas concentrations of other 06 constituents were less than 3.7%. Phenyl ethyl alcohol, lion shared constituent of essential oil was found highest (48.17%) in R. Gruss-an-Teplitz whereas minimum level was recorded in R. damascena (28.84%) under CW. In contrast, citronellol (chief component of fragrance) was highest in R. damascena (41.11%) in UTWW while the lowest level was found in R. Gruss-an-Teplitz (17.41%) in CW. This study confirmed the variations in metal concentrations of Rosa species due to different absorbability of each metal in flower petals. It also indicates that wastewater did not affect the composition but there were quantitative differences in aroma constituents and chemical composition of essential oil.

Heavy metal accumulation imparts structural differences in fragrant Rosa species irrigated with marginal quality water.

Wastewater is an alternative to traditional sources of renewable irrigation water in agriculture, particularly in water-scarce regions. However, the possible risks due to heavy metals accumulation in plant tissues are often overlooked by producers. The present study aimed to identify heavy metals-induced structural modifications to roots of scented Rosa species that were irrigated with water of marginal quality. The chemical and mineral contents from the experimental irrigation canal water (control) and treated wastewater were below the limits recommended by the Pakistan Environmental Protection Agency (Pak-EPA) for medicinal plants. The experimentally untreated wastewater contained electrical conductivity (EC), chemical oxygen demand (COD), biological oxygen demand (BOD), and heavy metals (Co, Cu, Cd, Pb) that were above the recommended limits. The responses by wastewater-treated Rosa species (Rosa damascena, R. bourboniana, R. Gruss-an-Teplitz, and R. centifolia) were evaluated. The experimental data revealed that treated wastewater significantly increased the thickness of collenchyma (cortex and pith) and parenchyma tissues (vascular bundle, xylem, and phloem) of R. Gruss-an-Teplitz. Root dermal tissues (epidermis) of R. bourboniana also responded to treated wastewater. R. damascena and R. centifolia were the least affected species, under the experimental irrigation conditions. Collenchyma and dermal tissues were thicker in R. damascena and R. Gruss-an-Teplitz under untreated wastewater conditions. In parenchyma tissues, vascular bundles were thicker in R. damascena in untreated wastewater conditions, while the xylem and phloem of R. Gruss-an-Teplitz were thicker where treated wastewater was applied. In tissues other than the vascular bundle, the differences in anatomical metrics due to the experimental irrigation treatments were greater during the second year of the experiment than in the first year. The contents of metals other than chromium in the roots and stems of roses were below the WHO limits, under all of the experimental irrigation conditions. Rosa centifolia contained higher heavy metals content than the other experimental species, and heavy metals content was associated with anatomical changes due to the treatments. We conclude that, under conditions of wastewater irrigation, R. Gruss-an-Teplitz was highly resistant; R. damascena was moderately resistant while R. bourboniana and R. centifolia were the most susceptible to irrigation with marginal quality water. This is the first report of plant tissue responses to wastewater irrigation by the experimental species. Regarding the accumulation of heavy metals in rose plant tissues, the results confirm that untreated wastewater must be treated to grow Rosa species where water is scarce.

Floral Characterization of Pomegranate Genotypes to Improve Hybridization Efficiency.

Pomegranate (Punica granatum) has staminate (male), androgynous (hermaphrodite), and intermediate flower types. Floral characterization is difficult for breeding efficiency across many pomegranate genotypes in Pakistan, which is essential for pomegranate cultivar enhancements. The present research focused on the floral characterization and breeding efficiency of fifteen pomegranate genotypes. Flower sex ratio, floral morphological parameters, i.e., flower length, ovary width, flower notch, flower tip and stigma with style length, and fruit set percentage were examined during the experiment. In terms of sex ratio, male flowers were found to be higher among all genotypes. Due to clear differences in flower length, width, and heterostyly facilitating visual identification of the hermaphrodite flowers, genotype Ternab-2, Kandhari White, and Kandhari Red had higher fruit set (≥70%) among all cross combinations attempted. Genotype Sava had higher flower length and heterostyly of hermaphrodite flower type, but ovary width was not very distinct, leading to average crossing success (85-34%). In conclusion, single or combination of morphological characters can be used for accurate identification of hermaphrodite flowers, which can improve hybrid efficiency and fruit set after artificial cross-pollination.

Genome Wide Analysis of Citrus sinensis Heat Shock Proteins.

BACKGROUND: Plant and animal cells possess a ubiquitous protein known as heat shock proteins (HSPs). Hsps were originally described in relation to heat shock and against abiotic and biotic stresses. Heat shock protein was classified in other crops on the bases of single classes or all classes but in Citrus sinensis Hsps groups, classes, subfamilies and members were not classified and characterized up to our knowledge. OBJECTIVES: Present study was focused on the identification and grouping of C. sinensis Hsps (CsHsps) classes, members among classes, their phylogenetic relationship, gene structure, conserved motifs and identification of proteins by using bioinformatics tools and analyses. MATERIALS AND METHODS: Genomic, Peptide and CDS sequences of CsHsps were downloaded from phytozome. MEGA 7 used for the phylogenetic analysis, GSDS for gene structure, UGENE for the multiple sequence alignment and MEME suite for the conserved motif analysis. RESULTS: The genome size of C. sinensis was 367 Mb, Chromosome number (2n)18, having 151 Hsps with six groups CsHsp10, 20, 40, 60,70 and 90. CsHsp20 was the largest group having 54 members, followed by CsHsp60 and CsHsp70 both having 30 members respectively. CONCLUSION: CsHsps members within a class shared more similar gene and protein structure. CsHsp 60, CsHsp 70 and CsHsp90 shared more conserved and similar amino acid pattern. Each class had some important proteins such as Cpn in CsHsp10, Hypothetical proteins in CsHsp20 and 40, Dnak in CsHsp60, Molecular chaperone in CsHsp70 and Hsp90 in CsHsp90. These proteins are produced by cells in response to stresses in citrus. Chaperonins and some hypothetical proteins identified in CsHsps, help in ATP synthesis and protein degradation. This is genome wide analysis and classification sets the groundwork for future investigations to fully characterize functionally the Citrus Hsps families and underscores the relevance of Hsps response to abiotic and biotic stresses in Citrus.

Effect of Antioxidants, Amino Acids and Plant Growth Regulators on in vitro Propagation of Rosa centifolia.

BACKGROUND: Rosa centifoliais commercially propagated by asexual means but in vitro propagation ensure the production of disease free and healthy plants and browning of explants creates hurdle in their multiplication. OBJECTIVES: The aim was to reduce oxidative browning of shoots of R. centifolia in MS medium during in vitro propagation. MATERIALS AND METHODS: Axillary buds of R. centifolia were sterilized with 70% ethyl alcohol for 4 min and 5% sodium hypochlorite for 2 min followed by three washing with sterilized double distilled water. In order to control oxidative browning, Ascorbic acid (100 mg.L-1), citric acid (100 mg.L-1) and activated charcoal (3 g.L-1) were used while to control withering of shoots, different concentrations (3.0 mg.L-1, 6.0 mg.L-1, 9.0 mg.L-1) of either glutamine, asparagine and proline were put into trial. Different concentrations of Benzyl aminopurine (BAP) and naphthalene acetic acid (NAA) were used for in vitro shoot and root formation. RESULTS: Minimum browning percentage (20%) was achieved in the presence of activated charcoal (3.0 g.L-1) and pretreatment of explants with running tap water. Asparagin (9.0 mg.L-1) produced maximum shooting (93%), minimum withering (6.67%), and it took longer period (27 days) for shoots to wither. BAP (3.0 mg.L-1) + NAA (0.5 mg.L-1) was produced the highest number of shoots (1.63), in a shortest periods (9 days). For root production, NAA (1.5 mg.L-1) + BAP (0.5 mg.L-1) reduced the time to 11 days with maximum number of roots (4.33) and root length (4.20 cm). CONCLUSIONS: The supplement of activated charcoal (3.0 g.L-1), a sparagin (9.0 mg.L-1) and combination of BAP and NAA in the MS medium is effective for in vitro propagation of R. centifolia.

Development of molecular method for sex identification in date palm (Phoenix dactylifera L.) plantlets using novel sex-linked microsatellite markers.

Microsatellite markers containing simple sequence repeats (SSRs) are a valuable tool for genetic analysis. Date palm is a dioecious and slow flowering and is very difficult to identify the gender of the trees until it reaches the reproductive age (5-10 years). A total of 12 microsatellite primers were used with 30 date palm samples, 14 parents (8 male + 6 females) and 16 progeny (developed from parents breeding) which showed that microsatellites were highly polymorphic, having a great number of alleles. A total of 124 alleles were characterized in 12 SSR loci. On average, there are 9.08 alleles per locus, with a range from 5 to 16 alleles, for primers mpdCIR15 and mpdCIR57, respectively. These primers produced 15 polymorphic loci specifically in male date palm samples and the seedlings harboring the unique fragments were further characterized as male plants. Increasingly, 38.46 % of these loci were scored as homozygous alleles while 61.53 % heterozygous allelic loci were determined. Primer mpdCIR48 produced a specific locus (250/250) in all male samples whereas the same locus was absent in female samples. Similarly, a locus of 300/310 bp reoccurred in 5 date palm male samples using marker DP-168 which indicated that these are the promising candidate marker to detect the sex in date palm seedlings at early stage. The data resulted from combination of 12 primers enabled the 16 seedling samples progeny (developed from parents breeding) of date palm cultivars to divide into two groups i.e., male and female regarding their sex expression comparative to the parents (male + female) using the principle coordinate analysis.