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In the present study an effort has been made to optimize the in vitro regeneration protocol for Agrobacterium-mediated transformation of Brassica juncea, because of its importance as oilseed crops. The highest callus induction frequency of 87% was observed on MS (Murashige and Skoog, 1962) medium supplemented with 4 µM 6-benzyladenine (BA) after four weeks of culture period. Subculturing of organogenic calli in MS media with a similar hormonal composition resulted in shoot organogenesis after six weeks of culture cultivation. The highest shoot induction frequency (92%) was recorded on MS medium containing 4 µM BA in combination with 1 µM of α-naphthalene acetic acid (NAA). Further, well-developed roots were formed in MS media augmented with 6 µM of Indole acetic acid (IAA) in combination with 1 µM Kinetin (Kn). Cotyledon explants were exploited in vitro for the successful transformation of B. juncea. A binary vector comprised of the Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene under the transcriptional control of a glycinin promoter and with a basta selection marker was introduced into A. tumefaciens strain GV3101 via electroporation. EaDAcT gene is responsible for unusual triacylglycerol’s production where the sn-3 position is esterified with acetate instead of the long-chain fatty acid found in the triacylglycerol’s. The highest regeneration frequency (100%) of transgenic shoots was observed on MS medium supplemented with 4 µM BA plus 1 µM NAA in the presence of 25 mg l−1 basta and 160 mg l−1 timintin. The efficiency of stable transformation was found to be approximately 7% in the transgenic plants. Moreover, the transformed regenerated shoots were confirmed by PCR analysis using EaDAcT gene-specific primers. Keywords: Brassica juncea, Callus, Agrobacterium tumefaciens, Transgenic plants, Biodiesel

Background: Germin-like Proteins (GLPs) play an important role in various stresses. Rice contains 43 GLPs, among which many remain functionally unexplored. The computational analysis will provide significant insight into their function. Objective: To find various structural properties, functional importance, phylogeny and expression pattern of all OsGLPs using various bioinformatics tools. Methods: Physiochemical properties, sub-cellular localization, domain composition, Nglycosylation and Phosphorylation sites, and 3D structural models of the OsGLPs were predicted using various bioinformatics tools. Functional analysis was carried out with the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) and Blast2GO servers. The expression profile of the OsGLPs was predicted by retrieving the data for expression values from tissuespecific and hormonal stressed array libraries of RiceXPro. Their phylogenetic relationship was computed using Molecular and Evolutionary Genetic Analysis (MEGA6) tool. Results: Most of the OsGLPs are stable in the cellular environment with a prominent expression in the extracellular region (57%) and plasma membrane (33%). Besides, 3 basic cupin domains, 7 more were reported, among which NTTNKVGSNVTLINV, FLLAALLALASWQAI, and MASSSF were common to 99% of the sequences, related to bacterial pathogenicity, peroxidase activity, and peptide signal activity, respectively. Structurally, OsGLPs are similar but functionally they are diverse with novel enzymatic activities of oxalate decarboxylase, lyase, peroxidase, and oxidoreductase. Expression analysis revealed prominent activities in the root, endosperm, and leaves. OsGLPs were strongly expressed by abscisic acid, auxin, gibberellin, cytokinin, and brassinosteroid. Phylogenetically they showed polyphyletic origin with a narrow genetic background of 0.05%. OsGLPs of chromosome 3, 8, and 12 are functionally more important due to their defensive role against various stresses through co-expression strategy. Conclusion: The analysis will help to utilize OsGLPs in future food programs.

In the present study an effort has been made to optimize the in vitro regeneration protocol for Agrobacterium-mediated transformation of Brassica juncea, because of its importance as oilseed crops. The highest callus induction frequency of 87% was observed on MS (Murashige and Skoog, 1962) medium supplemented with 4 µM 6-benzyladenine (BA) after four weeks of culture period. Subculturing of organogenic calli in MS media with a similar hormonal composition resulted in shoot organogenesis after six weeks of culture cultivation. The highest shoot induction frequency (92%) was recorded on MS medium containing 4 µM BA in combination with 1 µM of α-naphthalene acetic acid (NAA). Further, well-developed roots were formed in MS media augmented with 6 µM of Indole acetic acid (IAA) in combination with 1 µM Kinetin (Kn). Cotyledon explants were exploited in vitro for the successful transformation of B. juncea. A binary vector comprised of the Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene under the transcriptional control of a glycinin promoter and with a basta selection marker was introduced into A. tumefaciens strain GV3101 via electroporation. EaDAcT gene is responsible for unusual triacylglycerol’s production where the sn-3 position is esterified with acetate instead of the long-chain fatty acid found in the triacylglycerol’s. The highest regeneration frequency (100%) of transgenic shoots was observed on MS medium supplemented with 4 µM BA plus 1 µM NAA in the presence of 25 mg l−1 basta and 160 mg l−1 timintin. The efficiency of stable transformation was found to be approximately 7% in the transgenic plants. Moreover, the transformed regenerated shoots were confirmed by PCR analysis using EaDAcT gene-specific primers.

Clinically, available synthetic chemotherapeutics in the treatment for leishmaniasis are associated with serious complications, such as toxicity and emergence of resistance. Natural products from plants can provide better remedies against the Leishmania parasite and can possibly minimize the associated side effects. In this study, various extracts of the callus cultures of Artimisia scoparia established in response to different plant growth regulators (PGRs) were evaluated for their anti-leishmanial effects against Leishmania tropica promastigotes, followed by an investigation of the possible mechanism of action through reactive apoptosis assay using fluorescent microscopy. Amongst the different callus extracts, higher anti-leishmanial activity (IC

A field experiment was carried out at the Agriculture Research Institute Tarnab during the year. The response of peach nursery plants to different doses of nitrogen and plant densities was studied. Three levels of nitrogen, 0, 57 and 114 kg/hectare were used along with four different planting densities i.e. 536, 804, 1005 and 1072 seedlings per plot. It was found that seedling height (cm), diameter (mm), bud take success (%), scion length (cm) and root volume (cm­­­3) were significantly affected by different levels of nitrogen and planting densities, whereas time taken to reach buddable size was not affected in any treatment. Maximum seedling height, diameter, bud take success, scion length and root volume were 94.34 cm, 7.17 mm, 82.43%, 181.25cm and 667cm3 respectively for the maximum nitrogen dose of 114 kg ha-1. For different plant densities, the maximum seedling height, diameter, bud take success, scion length and root volume were 94.12 cm, 7.16 mm, 79.85%, 178.34 cm, and 672 cm3 respectively, for plots having 506 plants. Though, statistically the seedling height (98.51 cm), seedling diameter (8.17 mm), scion length (187.25 cm) and root volume (783 cm3) were highest for interaction, in plots with 114 kg nitrogen ha-1 and 536 seedlings. But the seedling height (95.24 cm), seedling diameter (7.12 mm), scion length (182.71 cm) and root volume (746 cm3) in the plots receiving the similar nitrogen dose having plant density of 804 were found to be comparably more economical. Thus, from our findings we recommend a plant density of 85760 and a nitrogen dose of 114 kg per hectare. Keywords: Peach (Prunus persica L.); Nitrogen; Seedling height; Seedling diameter; Root volume http://dx.doi.org/10.19045/bspab.2017.60056

To elucidate how biosynthesis of plant metabolites is affected by temperature, metabolite profiles from in vitro regenerated plants raised under different temperature regimes of 10, 15 °C, 20 °C, 25 °C and 30 °C were obtained using electrospray ionization mass spectrometry (ESI-MS), and principal component analysis (PCA) was carried out to identify key metabolites. Several bin masses were detected by PCA loading scatter plots which separated the samples. In-house bin program selectively manifested the putative known metabolites depending on % total ions count and intensity of selected bins in the plant samples. Total phenolic and flavonoid content were harvested to highest levels (12.9 mg GAE/g DW and 9.3 mg QE/g DW), respectively, at 15 °C. Besides, pinoresinol (lignan), some of the vital amino acids such as serine, methionine, histidine and glutamine were found to be at higher amount in plants raised at 15 °C. Significant phenylpropanoids like cinnamic acid, caffeic acid and quercitol were detected at a higher concentration in plants raised at 15 °C as compared to other treatments. However, phosphoenolpyruvate, and oxalosuccinate (intermediates of the pentose phosphate pathway) were accumulated the most in plants raised at 30 °C and they were detected with lowest values at 10 °C. Glucose and deoxy-xylose 5 phosphate (intermediates of TCA cycle) were found in higher amounts at temperature treatments of 15 and 25 °C, respectively. We conclude that a low-temperature treatment (15 °C) results in a stress-induced accumulation of a variety of pharmacologically important secondary metabolites.

Brassica juncea germplasm was evaluated to estimate divergence, associations among traits and heritability for various bio-chemical traits in RCB design having three replications at the University of Agriculture, Peshawar during 2011-12. Data were recorded on six bio-chemical traits viz., oil content, erucic acid content, glucosinolate content, protein content, oleic acid content and linolenic acid content. Highly significant variations were recorded for all the studied bio-chemical traits. Maximum heritability was observed for glucosinolate content (0.90), protein content (0.80) and erucic acid content (0.82), whereas moderate heritability was recorded for oil content (0.54), oleic acid content (0.33) and linolenic acid (0.43). The high heritability was coupled with appreciable selection response. Negative associations were recorded among oil content and protein content (r= -0.419**). Protein content showed positive associations with glucosinolate content (r= 0.678**) and erucic acid content (r= 0.481**). Keywords: Brassica Juncea L.; Heritability; Correlation; Selection response http://dx.doi.org/10.19045/bspab.2016.50168

An experiment was conducted at Agricultural Research Station Buner in summer 2014 to study the effect of plowing time i.e. in dark (at night) and in sunlight (at day) and sowing time i.e. immediate after plowing and 12 hours later, on various maize (Zea mays L.) hybrids. For this purpose, two fields were selected, one for daytime plowing and other for night time plowing. Night plowing was done at 10.00 p.m. while daytime plowing at 8.00 a.m. Furthermore; half of the plowed field was sown immediately after plowing and the rest, 12 hours later in both plowing treatments. Three hybrids of maize i.e. Monsanto-2021, Rafhan-3313 and Rafhan-3304 were planted in each field under randomized complete block design with four replications. All standard agronomic practices were adopted during the experiment for all treatments. All the observed parameters were significantly affected by plowing time, while sowing time had no impact. Generally, a significant interaction among plowing and sowing time was observed for the recorded parameters. Early emergence (7 days) was observed for the field plowed at night. Plowing the field at usual daytime delayed emergence (9 days). Delayed tasseling (52.9 days), delayed silking (58.6 days),largerleaf area (850 cm2), delayed maturity (114 days), taller plants (271cm), greater stover yield (12328 kg ha-1) and accelerated crop growth rate (24.98 gm-2.day-1) were recorded for the field plowed at night. There were also significant differences among various hybrids, where Monsanto-2021 delivered better results. Plowing the field at night proved superior in term of crop growth rate as compared to the field plowed at usual daytime. Night plowing is recommended to increase vegetative performance of maize. Sowing can be done immediately after night plowing or on the subsequent day of plowing. Hybrids particularly, Monsanto-2021 is recommended for better performance. Keywords: Maize; Night plowing; Monsanto-2021; Rafhan-3313 and Rafhan-3304 http://dx.doi.org/10.19045/bspab.2017.60005

To support the crop improvement of Physalis ixopcarpa Brot., 16 accessions provided by National Plant Germ Plasm System USDA (USA) and one recently reported accession from Pakistan were characterized for morphometric and genetic diversity. These accessions were evaluated for morphometric and physiological traits. Among these traits, plant height, relative water content, total chlorophyll content, per cent germination and leaf area were found as best for the differentiation among these accessions. These accessions were also screened for genetic differentiation using RAPDs primers. The genetic similarity among accessions was in the range of 0.03 to 0.33. The RAPD data supported the ecological distribution of accessions. It is inferred from molecular typing that accessions PI512005 and PI360740 were the most diverse accessions among the accessions of P. ixocarpa. In this study the genetic data were deviated from morphometric data which might be due to the qualitative and quantitative characteristics of accessions influenced by environmental factors.