Sumera Afzal Khan

Plant growth promotion and Penicillium citrinum.

BackgroundEndophytic fungi are known plant symbionts. They produce a variety of beneficial metabolites for plant growth and survival, as well as defend their hosts from attack of certain pathogens. Coastal dunes are nutrient deficient and offer harsh, saline environment for the existing flora and fauna. Endophytic fungi may play an important role in plant survival by enhancing nutrient uptake and producing growth-promoting metabolites such as gibberellins and auxins. We screened roots of Ixeris repenes (L.) A. Gray, a common dune plant, for the isolation of gibberellin secreting endophytic fungi.ResultsWe isolated 15 endophytic fungi from the roots of Ixeris repenes and screened them for growth promoting secondary metabolites. The fungal isolate IR-3-3 gave maximum plant growth when applied to waito-c rice and Atriplex gemelinii seedlings. Analysis of the culture filtrate of IR-3-3 showed the presence of physiologically active gibberellins, GA1, GA3, GA4 and GA7 (1.95 ng/ml, 3.83 ng/ml, 6.03 ng/ml and 2.35 ng/ml, respectively) along with other physiologically inactive GA5, GA9, GA12, GA15, GA19, GA20 and, GA24. The plant growth promotion and gibberellin producing capacity of IR-3-3 was much higher than the wild type Gibberella fujikuroi, which was taken as control during present study. GA5, a precursor of bioactive GA3 was reported for the first time in fungi. The fungal isolate IR-3-3 was identified as a new strain of Penicillium citrinum (named as P. citrinum KACC43900) through phylogenetic analysis of 18S rDNA sequence.ConclusionIsolation of new strain of Penicillium citrinum from the sand dune flora is interesting as information on the presence of Pencillium species in coastal sand dunes is limited. The plant growth promoting ability of this fungal strain may help in conservation and revegetation of the rapidly eroding sand dune flora. Penicillium citrinum is already known for producing mycotoxin citrinin and cellulose digesting enzymes like cellulase and endoglucanase, as well as xylulase. Gibberellins producing ability of this fungus and the discovery about the presence of GA5 will open new aspects of research and investigations.

Exogenous gibberellic acid reprograms soybean to higher growth and salt stress tolerance.

The agricultural industry is severely affected by salinity due to its high magnitude of adverse impacts and worldwide distribution. We observed the role of exogenous gibberellic acid (GA(3)) in salinity alleviation of soybean. We found that GA(3) application significantly promoted plant length and plant fresh/dry biomass while markedly hindered by NaCl induced salt stress. The adverse effect of salt stress was mitigated by GA(3), as growth attributes significantly recovered, when GA(3) was added to salt stressed soybean plants. Elevated GA(3) treatments increased daidzein and genistein contents (commonly known as phytoestrogens) of soybean leaves under control and salt stress conditions. Phytohormonal analysis of soybean showed that the level of bioactive gibberellins (GA(1) and GA(4)) and jasmonic acid increased in GA(3) treated plants, while the endogenous abscisic acid and salicylic acid contents declined under the same treatment. GA(3) mitigated the adverse effects of salt stress by regulating the level of phytohormones, thus aiding the plant in resuming its normal growth and development. The presence of GA(1) and GA(4) showed that both early-C13-hydroxylation and non-C13-hydroxylation pathways of GA biosynthesis are functional in soybean. It was concluded that GA(3) ameliorates the adverse effects of salt stress and restores normal growth and development of soybean.

Pure culture of Metarhizium anisopliae LHL07 reprograms soybean to higher growth and mitigates salt stress

Little is known about the role of endophytic fungi against abiotic stresses and isoflavonoids (IF) contents of soybean. In current study, we investigated the role of fungal endophytes on the growth of soybean under salt stress conditions. Pure cultures of nine endophytic fungi were isolated from the roots of field-grown soybean plants, and their culture filtrates were screened on Waito-C and Dongjin-byeo rice cultivars; for identification of plant growth promoting fungal strains. It was observed that fungal isolate GMC-2B significantly promoted the growth of both Waito-C and Dongjin-byeo. GMC-2B was later identified as a new strain of Metarhizium anisopliae LHL07 on the basis of 18S rDNA sequences and phylogenetic analysis. Metarhizium anisopliae LHL07 inoculated soybean plants recorded significantly higher shoot length, shoot fresh and dry biomass, chlorophyll contents, transpiration rate, photosynthetic rate and leaf area; under sodium chloride induced salt stress as compared to non-inoculated control plants. An elevated proline and reduced superoxide dismutase and malondialdehyde contents in M. anisopliae LHL07 inoculated soybean plants demonstrated mitigation of salt induced oxidative stress. Furthermore, reduced abscisic acid and elevated jasmonic acid contents in soybean plants confirmed that lesser stress was convened to M. anisopliae inoculated-plants under salinity stress. We also assessed the role of M. anisopliae interaction on IF biosynthesis of soybean, and found significantly higher IF contents in M. anisopliae inoculated soybean plants. In conclusion, endophytic fungal interactions with soybean can be beneficial to improve soybean quality and quantity under salt affected agricultural systems.

Gibberellin production and plant growth promotion from pure cultures of Cladosporium sp. MH-6 isolated from cucumber (Cucumis sativus L.)

Gibberellin (GA) production by soil fungi has received little attention, although substantial work has been carried out on other aspects of plant growth promoting fungi (PGPF). In our studies we investigated GA production and growth-promoting capacity of a novel fungal strain isolated from the roots of soil-grown cucumber. Pure cultures of 19 endophytic fungi were tested for shoot length promotion of Waito-C rice to identify the GA production capacity of these fungal isolates. Isolate MH-6 significantly increased shoot length (12.9 cm) of Waito-C, in comparison to control treatments. Bioassay with culture filtrate (CF) of MH-6 also significantly promoted growth attributes of cucumber plants. Analysis of MH-6 CF showed the presence of physiologically active (GA1, 1.97 ng/mL; GA3, 5.18 ng/mL; GA4, 13.35 ng/mL and GA7, 2.4 ng/ mL) in conjunction with physiologically inactive (GA9 [0.69 ng/mL], GA12 [0.24 ng/mL], GA15 [0.68 ng/mL, GA19 [1.94 ng/mL and GA20 [0.78 ng/mL]) gibberellins. The CF of MH-6 produced greater amounts of GA3, GA4, GA7 and GA19 than wild type Fusarium fujikuroi, a fungus known for high production of GA. The fungal isolate MH-6 was identified as a new strain of Cladosporium sp. on the basis of sequence homology (99%) and phylogenetic analysis of 18S rDNA sequence.

Chrysosporium pseudomerdarium produces gibberellins and promotes plant growth.

We isolated 10 endophytic fungi from the roots of drought stressed soybean cultivar Hwangkeumkong and bioassyed on waito-c rice and soybean seedlings, in order to identify plant growth-promoting fungi. The fungal isolate D-2-1 provided the best result for plant height and biomass promotion as compared to wild type Gibberella fujikuroi. The D-2-1 culture filtrate (CF) was analyzed for the presence of gibberellins (GAs) and it was observed that all physiologically active GAs, especially gibberellic acid, were present in higher amounts (GA1, 0.24 ng/ml; GA3, 8.99 ng/ml; GA4, 2.58 ng/ml and GA7, 1.39 ng/ml) in conjunction with physiologically inactive GA5, GA9, GA15, GA19, and GA24. The fungal isolate D-2-1 was identified as a new strain of Chrysosporium pseudomerdarium through phylogenetic analysis of 18S rDNA sequence. Plant growth promotion and GAs production capacity of genus Chrysosporium have been reported for the first time in this study.

Gibberellins Producing Endophytic Fungus Porostereum spadiceum AGH786 Rescues Growth of Salt Affected Soybean

In the pursuit of sustainable agriculture through environment and human health friendly practices, we evaluated the potential of a novel gibberellins (GAs) producing basidiomycetous endophytic fungus Porostereum spadiceum AGH786, for alleviating salt stress and promoting health benefits of soybean. Soybean seedlings exposed to different levels of NaCl stress (70 and 140 mM) under greenhouse conditions, were inoculated with the AGH786 strain. Levels of phytohormones including GAs, JA and ABA, and isoflavones were compared in control and the inoculated seedlings to understand the mechanism through which the stress is alleviated. Gibberellins producing endophytic fungi have been vital for promoting plant growth under normal and stress conditions. We report P. spadiceum AGH786 as the ever first GAs producing basidiomycetous fungus capable of producing six types of GAs. In comparison to the so for most efficient GAs producing Gibberella fujikuroi, AGH786 produced significantly higher amount of the bioactive GA3. Salt-stressed phenotype of soybean seedlings was characterized by low content of GAs and high amount of ABA and JA with reduced shoot length, biomass, leaf area, chlorophyll contents, and rate of photosynthesis. Mitigation of salt stress by AGH786 was always accompanied by high GAs, and low ABA and JA, suggesting that this endophytic fungus reduces the effect of salinity by modulating endogenous phytohormones of the seedlings. Additionally, this strain also enhanced the endogenous level of two isoflavones including daidzen and genistein in soybean seedlings under normal as well as salt stress conditions as compared to their respective controls. P. spadiceum AGH786 boosted the NaCl stress tolerance and growth in soybean, by modulating seedlings endogenous phytohormones and isoflavones suggesting a valuable contribution of this potent fungal biofertilizer in sustainable agriculture in salt affected soils.

Genotyping of HCV RNA Reveals That 3a Is the Most Prevalent Genotype in Mardan, Pakistan

The clinical outcomes of patients infected with hepatitis C virus (HCV) range from acute resolving hepatitis to chronic liver diseases such as liver cirrhosis or hepatocellular carcinoma. Identification of the infecting virus genotype is indispensable for the exploration of many aspects of HCV infection, including epidemiology, pathogenesis, and response to antiviral therapy. 1419 individuals were screened for anti-HCV in this study, of which 166 (11.7%) were found reactive by ICT (Immunochromatographic test). These 166 anti-HCV positive and 26 normal individuals were further analyzed. RNA was extracted from serum and reverse-transcribed to cDNA and the core region of HCV genome was targeted and amplified by multiplex PCR. HCV RNA was detected in 121 individuals, of which 87 were male and 34 were female. Genotype 3a was the most prevalent among all the genotypes observed followed by 3b. Genotypes 1a, 2a, and 2b were found in 10.89%, 13.22%, and 6.61% patients, respectively. 25.41% of the HCV RNA positive samples were not typed. 6.05% of patients were found having mixed genotypes. These findings will not only help the physicians to prescribe more appropriate treatment for the HCV infection but will also draw the attention of health-related policy makers to devise strategies to curb the disease more effectively.

Effects of Endophytic Fungal Secondary Metabolites on the Growth and Physiological Response of Carex kobomugi Ohwi

Abstract Recently, plant growth–promoting endophytic fungi with gibberellins-producing ability were isolated from the roots of coastal sand dune plants. Carex kobomugi Ohwi in coastal sand dunes was treated with the supernatant solution of Penicillium citrinum KACC43900 isolated from Ixeris repens (L.) A. Gray. After treatment, the length of leaf blades, photosynthesis, chlorophyll fluorescence, and soil respiration were measured in the field. The contents of chlorophyll and carotenoid were determined with leaf blades selected for measurement of chlorophyll fluorescence. Fungal culture filtrate (FCF) increased leaf blade length, the contents of chlorophyll a (Chl a) and chlorophyll b (Chl b), total chlorophyll (total Chl [a + b]), and carotenoid in leaf blades of C. kobomugi. FCF also increased net photosynthetic rate (A), transpiration rate (E), carboxylation efficiency (A/Ci), and water-use efficiency (A/E). However, the ratio of variable fluorescence to maximal fluorescence (Fv/Fm) did not show significant difference between control and FCF-treated plants. Soil respiration rates were higher in the site treated with FCF than the control. Although growth response and physiological characteristics of C. kobomugi were different according to the treatment methods of FCF, injection or spray treatment, plant growth of C. kobomugi was increased with FCF treatment.

Kinetin modulates physio-hormonal attributes and isoflavone contents of Soybean grown under salinity stress

Crop productivity continues to decline due to a wide array of biotic and abiotic stresses. Salinity is one of the worst abiotic stresses, as it causes huge losses to crop yield each year. Kinetin (Kn) has been reported as plant growth regulator since long, but its role in improving plant growth and food quality under saline conditions through mediating phytohormonal cross-talk is poorly studied. Current study was designed to evaluate the impact of exogenously applied Kn on growth, isoflovones and endogenous phytohormones of soybean grown under NaCl induced salt stress. Soybean plants were grown in perlite (semi hydroponic), and under controlled green-house conditions. Elevated levels of exogenous Kn significantly mitigated the adverse effect of NaCl and rescued plant growth attributes, i.e., plant height, fresh and dry biomass of soybean plants grown in all treatments. Higher diadzen, glycitin, and genistin contents were observed in plants treated with elevated Kn in the presence or absence of NaCl induce salt stress. The gibberellins (GAs) biosynthesis pathway was up-regulated by Kn as the bioactive GA1 and GA4 contents were significantly higher in Kn treated plants, as compared to control, while GAs level reduced in NaCl treated plants. Contrary to GAs, the abscisic acid contents declined with Kn but promoted in NaCl stressed soybean plants. The endogenous jasmonic acid and salicylic acid contents of soybean enhanced with elevated Kn application, but they showed an antagonistic response under salt stress. Current study supports the active role of Kn to ameliorate the adverse effects of salt stress on the growth and food quality of soybean. The favorable role of Kn toward soybean growth under salt stress may be attributed to its potential to modulate cross-talk between the various phytohormones involved in soybean growth and its resistance to salinity stress.

STUDIES ON DIABETIC NEPHROPATHY AND SECONDARY DISEASES IN TYPE 2 DIABETES

Present investigations include 548 diabetes mellitus patients of which 311 (56.75%) were male and 237 (43.25%) were female patients. Type 2 nephropathy patients totalled 353(64.42%) of which 196(55.52%) were males and 157(44.48%) were females. Mean age at diagnosis of type 2 nephropathy patients was 61.64±0.48 years. Mean body mass index (BMI) estimated in type 2 nephropathy patients was 26.77±0.15 Kg/m 2 . Biochemical evaluations in type 2 nephropathy patients was as follows: mean blood urea 156.02±1.48mg/100ml, mean serum creatinine 6.91±0.06mg/100ml, mean blood glucose level 339.39±3.34mg/100ml, potassium 6.81±0.06 Mmol/Lit and mean sodium was 157.44±0.77Mmol/ Lit. Microalbuminuria was found in 43.63% and the remainder had macroalbuminuria. Secondary diseases associated with diabetic nephropathy in type2 nephropathy were retinopathy, hypertension, diabetic foot and neuropathy. The highest percentage of type 2 nephropathy (type2N) patients were those who had no school education and the lowest percentage was of those who had education of university level. As far as socio-economic status was concerned, the highest percentage (25.22%) of type2N patients was of skilled personals. Calculated coefficient of inbreeding (F) for type2N patients was 0.028.