Syed Abdullah Gilani

Secondary Metabolites from Inula britannica L. and Their Biological Activities

Inula britannica L., family Asteraceae, is used in traditional Chinese and Kampo Medicines for various diseases. Flowers or the aerial parts are a rich source of secondary metabolites. These consist mainly of terpenoids (sesquiterpene lactones and dimmers, diterpenes and triterpenoids) and flavonoids. The isolated compounds have shown diverse biological activities: anticancer, antioxidant, anti-inflammatory, neuroprotective and hepatoprotective activities. This review provides information on isolated bioactive phytochemicals and pharmacological potentials of I. britannica.

Fungal endophyte Penicillium janthinellum LK5 improves growth of ABA-deficient tomato under salinity

An endophytic fungus was isolated from the roots of tomato (Solanum lycopersicum Mill) and identified as Penicillium janthinellum LK5. The culture filtrate (CF) of P. janthinellum significantly increased the shoot length of gibberellins (GAs) deficient mutant waito-c and normal Dongjin-beyo rice seedlings as compared to control. The CF of P. janthinellum contained GAs (GA3, GA4, GA7 and GA12). To assess endophyte-growth promoting and stress-tolerance potential, the CF along with the propagules of endophyte was applied to tomato-host and abscisic acid (ABA)-deficient mutant Sitiens plants under sodium chloride (NaCl) induced salinity stress. Sitiens plants had retarded growth under normal and salinity stress however its growth was much improved during P. janthinellum-association. The endophyte inoculation reduced the membrane injury by decreasing lipid peroxidation as compared to non-inoculated control under salinity. Endophyte-associated Sitiens plants have significantly higher catalase, peroxidase and glutathione activities as compared to control. Endophyte-infected host and Sitiens plants had low level of sodium ion toxicity and high calcium contents in its root as compared to control. P. janthinellum LK5 helped the Sitiens plants to synthesis significantly higher ABA and reduced the level of jasmonic acid to modulate stress responses. The results suggest that endophytes-association can resist salinity stress by producing gibberellins and activating defensive mechanisms of host and Sitiens plants to achieve improved growth.

Endophytes from medicinal plants and their potential for producing indole acetic acid, improving seed germination and mitigating oxidative stress.

Medicinal plants have been used by marginal communities to treat various ailments. However, the potential of endophytes within these bio-prospective medicinal plants remains unknown. The present study elucidates the endophytic diversity of medicinal plants (Caralluma acutangula, Rhazya stricta, and Moringa peregrina) and the endophyte role in seed growth and oxidative stress. Various organs of medicinal plants yielded ten endophytes, which were identified as Phoma sp. (6 isolates), Alternaria sp. (2), Bipolaris sp. (1), and Cladosporium sp. (1) based on 18S rDNA sequencing and phylogenetic analysis. The culture filtrates (CFs; 25%, 50%, and 100% concentrations) from these endophytes were tested against the growth of normal and dwarf mutant rice lines. Endophytic CF exhibited dose-dependent growth stimulation and suppression effects. CF (100%) of Phoma sp. significantly increased rice seed germination and growth compared to controls and other endophytes. This growth-promoting effect was due to the presence of indole acetic acid in endophytic CF. The gas chromatography/mass spectrometry (GC/MS) analysis showed the highest indole acetic acid content ((54.31±0.21) μmol/L) in Bipolaris sp. In addition, the isolate of Bipolaris sp. exhibited significantly higher radical scavenging and anti-lipid peroxidation activity than the other isolates. Bipolaris sp. and Phoma sp. also exhibited significantly higher flavonoid and phenolic contents. The medicinal plants exhibited the presence of bio-prospective endophytic strains, which could be used for the improvement of crop growth and the mitigation of oxidative stresses.摘要目的探讨药用植物内生菌的多样性及其在种子生长和氧化应激中的作用。方法从三种药用植物(Caralluma acutangula、Rhazya stricta 和 Moringa peregrina)中提取内生菌;基 于18S rDNA 测序和系统发育分析鉴定分离得到 的内生菌株;以正常和矮化突变体水稻品系为对 照,比较不同浓度的内生菌培养滤液(CF)对水 稻种子的发芽和生长的影响;通过气相色谱-质谱 分析CF 中的有效活性成分。结论从药用植物中共获得10 种内生菌,包括茎点霉 属6 株、链格孢属2 株、双极霉属1 株和枝孢霉 属1 株。CF 表现出剂量依赖性的生长刺激和抑制 作用。与对照和其他内生菌相比,100%的茎点霉 菌CF 显著促进了水稻种子的发芽和生长;双极 霉中的吲哚乙酸含量最高,并表现出比其更高的 自由基清除和抗脂质过氧化活性;双极霉菌和茎 点霉菌的类黄酮和酚类成分较高。综上所述,药 用植物中存在内生菌株,其可以用于改善作物生 长和减轻氧化应激。

Elemental allelopathy and antifungal activities of Inula falconeri from Himalaya Pakistan

Abstract The leaf leachate of Inula falconeri Hook. f. has been reported to carry allelopathic potential. To analyse its further effects, it was fractionated into five subfractions, i.e. hexane, chloroform, ethyl acetate, n-butanol, and water. The hexane subfraction had the highest specific activity compared with the other four fractions and presented 100% inhibitory effect to germination of lettuce seeds. Chloroform subfraction presented a dose-response effect, and the ethyl acetate subfraction exhibited insignificant inhibitory effect, while n-butanol and water subfractions presented stimulatory effects. Elemental analysis through energy-dispersive X-ray spectroscopy (EDX) showed the composition of 15 different elements with carbon, oxygen, and potassium showing highest weight percentages. In the case of hexane subfraction, highest percentages of silver (1.22%), cadmium (1.44%), manganese (0.71%), and chromium (0.70%) were observed, compared with other subfractions. Chloroform subfraction has high proportions of copper (0.63%), nickel (0.45%), iron (0.37%), calcium (0.22%), and cobalt (0.08%). In the case of ethyl acetate subfraction oxygen (49.66%) and sodium (0.33%) percentage was the highest of all the subfractions, while the other elements were observed in lesser amounts than in other subfractions. The n-butanol subfraction has recorded highest weight of zinc (0.79%) compared with other subfractions while in water subfraction significant amounts of other elements were not observed. To investigate the behaviour of these subfractions against plant pathogens, hexane subfraction (concentration gradients of 1.25, 2.5, and 5 mg/disc) demonstrated significant zones of inhibition against Alternaria alternata and Rhizoctonia solani while only an insignificant effect against Fusarium oxysporum. Thus, using active natural inhibitors of plant pathogens reveals broader perspectives and interests in avoiding losses to cash crops in the local area.

Exploring the Potentials of Lysinibacillus sphaericus ZA9 for Plant Growth Promotion and Biocontrol Activities against Phytopathogenic Fungi

There is an ongoing hunt for biologically active compounds that can combat phytopathogenic fungi and improve plant growth without causing any hazards to the environment. Consequently the present study aims at deciphering the plant growth promotion and antifungal capability of Lysinibacillus sphaericus ZA9. The bacterium was previously isolated and identified in our laboratory from maize rhizosphere using 16S rRNA gene sequencing. The test bacterium L. sphaericus ZA9 was found to produce high quantity of IAA (697 μg/ mL); siderophores (195.79 μg/ mL), HCN and hydrolytic enzyme as compared to the reference strain Bacillus sphaericus Z2-7. The bacterium was also capable of solubilizing silicates (Si), phosphates (P), and potassium (K). The bacterium enhanced the seedling vigor and germination of seeds pretreated with it and promoted the shoot length of both cucumber and tomato seeds in greenhouse experiment. L. sphaericus ZA9 and its cell free culture supernatant showed varied antagonistic behavior against Alternaria alternata, Curvularia lunata, Aspergillus sp., Sclerotinia sp., Bipolaris spicifera, Trichophyton sp. Fermentation broth culture of L. sphaericus ZA9 was then used to isolate antifungal metabolites by silica column chromatography. Identification and determination of antifungal compounds was carried out by Thin-layer chromatography (TLC) followed by NMR spectroscopy. Two compounds were isolated and identified as 2-pentyl-4-quinolinecarboxylic acid (C15H17NO2) which is a quinoline alkaloid and 1- methylcyclohexene which is a cycloalkene. Compound 1; 2-Penthyl-4-quinolinecarboxylic acid was found to be highly antagonistic against most of the fungi tested as compared to the bacterium itself. Its activity was comparable to that of fungicide Benlate, while compound 2; 1- methylcyclohexene did not show any antifungal activity.

Morphological and molecular characterization of underutilized medicinal wild ginger ( Zingiber barbatum Wall.) from Myanmar

Zingiber barbatum Wall. (family Zingiberaceae), is an underutilized medicinal plant and commonly known as ‘Meik tha-lin’ in Myanmar where it is used in the indigenous system of medicine. In the present study, 19 accessions of Z. barbatum from five provinces in Myanmar have been utilized to characterize and assess genetic diversity. Twenty-nine morphological characters were noted, including growth habit, leaf, pseudo-stem and rhizome characters. Fifteen primer sets of P450-based analogue (PBA) markers were used to reveal molecular characteristics. Of the 29 morphological characters, 22 showed a high degree of variation within wild ginger accessions, whereas 20 of these characters contributed significantly to morphological variation. Eleven amplified primer sets gave a total of 175 bands and exhibited 92.15% polymorphism across Z. barbatum accessions. Based on morphological characters and PBA markers, 19 accessions can be divided into two morphotype groups with comparatively higher genetic diversity. This information can be applied in future crop improvement, proper conservation and better use of this underutilized medicinal species.

Effect of phosphate nutrition on growth, physiology and phosphate transporter expression of cucumber seedlings

Although abundantly present in soils, inorganic phosphate (Pi) acquisition by plants is highly dependent on the transmembrane phosphate transporter (PT) gene family. Cucumber (Cucumis sativus) requires a large amount of phosphorus (P). The purpose of this study was to isolate the CsPT2-1 from cucumber roots, and to determine the influence of Pi nutrition on cucumber growth, metabolism and transcript levels of CsPT2-1 in tissues. Full length CsPT2-1 was cloned and phylogenetically identified. In two greenhouse experiments, P-deficient seedlings provided with low or high P concentrations were sampled at 10 and 21 days post treatment, respectively. Addition of P dramatically reduced growth of roots but not shoots. Supplying plants with high P resulted in increased total protein in leaves. Acid phosphatase activity increased significantly in leaves at any rate higher than 4 mM P. Increasing P concentration had a notable decrease in glucose concentrations in leaves of plants supplied with >0.5 mM P. In roots, glucose and starch concentrations increased with increasing P supply. Steady-state transcript levels of CsPT2-1 were high in P-deprived roots, but declined when plants were provided >10 mM P. To our knowledge, this is the first report focusing on a PT and its expression levels in cucumber.

Detection and Quantification of Formalin Adulteration in Cow Milk Using Near Infrared Spectroscopy Combined with Multivariate Analysis

In order to increase the shelf life for long distance transportation of milk, formalin is added as an adulterant to milk. That is highly toxic causes liver and kidney damages. A new Near Infrared spectroscopy (NIR) combined with multivariate analysis was developed to detect as well as to quantify the level of formalin adulteration in cow milk. In this study four different types of cow milk samples were collected from Nizwa regions of Sultanate of Oman and were investigated. Those cow milk samples were then adulterated with formalin at eight different percentage levels: 0%, 1%, 3%, 5%, 7%, 9%, 11%, 13% and 17% of formalin. All samples were measured using NIR spectroscopy in absorption mode in the wavelength range from 700-2500 nm, at 2 cm-1 resolution and using a 0.2 mm path length CaF2 sealed cell. The multivariate methods like Principle component analysis (PCA), partial least discriminant analysis (PLS-DA) and partial least regression analysis (PLS) were applied for statistical analysis of the obtained NIR spectral data. PLS-DA model was used to check the discrimination between the pure and formalin adulterated milk samples. For PLSDA model the R-square value obtained was 0.969 with 0.086 RMSE (Root mean square error). Furthermore, PLS regression model was also built to quantify the levels formalin adulterant in cow milk samples. The PLS regression model was obtained with the R-square 93% and with 1.38 RMSECV(Root mean square error of cross validation) value having good prediction with RMSEP (Root mean square error of prediction) value 1.50 and correlation of 0.95. This newly developed method is non-destructive, cheap, no need of much sample preparation and having sensitivity level less than 2% level of formalin adulteration.

AFLP and PBA polymorphisms in an endangered medicinal plant, Rhazya stricta , in Pakistan

Rhazya stricta is an endangered medicinal plant that is under severe human pressure as a result of commercial harvesting. As a model of habitat fragmentation, we selected the Kohat Plateau in the Pakistani Himalaya, where populations are fragmented, with less than 100 individuals per population. We hypothesized that the populations may be threatened by rapid habitat fragmentation and by unsustainable utilization of the plant. We analysed P450-based analogue functional genomic markers and amplified fragment length polymorphism markers from six populations of R. stricta on the Kohat Plateau, and examined their variations both within and among the populations. Both the marker groups revealed lower genetic differentiation among the populations and higher genetic differentiation within the populations as a result of high gene flow. The results confirmed that habitat fragmentation is being caused by severe human pressure, and although signs of genetic erosion are not yet visible, they seem likely to become visible in the future. Therefore, the conservation of R. stricta populations is necessary.

New robust sensitive fluorescence spectroscopy coupled with PLSR for estimation of quercetin in Ziziphus mucronata and Ziziphus sativa

Flavonoids are natural antioxidants derived from plants and commonly found in a variety of foods to sequester free radicals. Quercetin, belonging to flavonol subclass of flavonoids, has received considerable attention because of its wide uses as a nutritional supplement as well as a phytochemical remedy for a number of diseases. In the current study, quantification of quercetin was carried out in two medicinally important flavonoid rich plant Ziziphus mucronata and Ziziphus sativa. Emission spectroscopy was utilized as a new method coupled with Partial Least Squares Regression (PLSR) and the cross validation was done by UV-Visible spectroscopy. The results indicated the higher quercetin content in Z. mucronata (1.50±0.034%) than Z. sativa (1.21±0.052%), and were further verified through Folin-Ciocalteu Colorimetric method (Z. mucronata; 1.41±0.26% and Z. sativa; 1.13±0.136%). In this study the sensitivity was explained in term of slope i.e. Slope=0.9973.