Abdul Samad Mumtaz

Medicinal plant diversity in the flora of Leepa Valley, Muzaffarabad (AJK), Pakistan

Plants are inevitable and indispensable in the life of mankind. They are a major source of nutrition and medicament in the world. In this regard, the Leepa valley (LV), Muzaffarabad Azad Jammu and Kashmir (AJK) was identified as the case study. The main objectives of this research were to explore and document the medicinal plant diversity (MPD) in the area. In this study, irrespective of gender, age, demography and literacy, 42 informants were approached and information was documented by using rapid rural appraisal (RRA) method comprising of open-ended and closed-ended interviews. Visual appraisal approach (VAA) was also performed as complimentary tool to support the data. In the study, some quantitative analysis tools: informant consensus factor (ICF), fidelity level (FL), priority ranking (PR) and data matrix ranking (DMR) were employed to find reliability and validity of methods used and informations obtained. This exploratory study revealed a profile of 36 medicinal plants (MPs) of 22 families with their botanic name, local name, family, plant part used, mode of medicament preparation, ethnomedicinal recipes and commercial importance. Family Pinaceae (4 spp.) was the first in the family index ranking followed by Lamiaceae (3 spp.). In health problems, snake bite and sting were the most prevailing (ICF: 0.94) and, hyperglycemia and hypertension were least encountered (ICF: 0.06) in the LV. Ajuga bracteosa (FL: 100%) and Berberis lycium (FL: 93%) were predominantly used in folklore therapies and FL analysis is good indicator for further pharmaceutical research taxa. Abies pindrow, Pinus wallichiana and Taxus wallichiana were found to be multifarious in use. Illicit export, incessant agriculture expansion and wooden house construction were determined as the major threats for MPD in the area. The pharmaceutical potential of MPD in drug discovery and development is discussed, and recommendations for sustainable use of MPs and their in situ and ex situ conservation is suggested for the socio-economic uplift of local communities.

L-asparaginase as a critical component to combat Acute Lymphoblastic Leukaemia (ALL): A novel approach to target ALL

L-asparaginase, an anti-leukaemic drug that has been approved for clinical use for many years in the treatment of childhood Acute Lymphoblastic Leukaemia (ALL), is obtained from bacterial origin (Escherichia coli and Erwinia carotovora). The efficacy of L-asparaginase has been discussed for the past 40 years, and an ideal substitute for the enzyme has not yet been developed. The early clearance from plasma (short half-life) and requirement for multiple administrations and hence frequent physician visits make the overall treatment cost quite high. In addition, a high rate of allergic reactions in patients receiving treatment with the enzyme isolated from bacterial sources make its clinical application challenging. For these reasons, various attempts are being made to overcome these barriers. Therefore, the present article reviews studies focused on seeking substitutes for L-asparaginase through alternative sources including bacteria, fungi, actinomycetes, algae and plants to overcome these limitations. In addition, the role of chemical modifications and protein engineering approaches to enhance the drugs efficacy are also discussed. Moreover, an overview has also been provided in the current review regarding the contradiction among various researchers regarding the significance of the enzymes glutaminase activity.

Documenting the indigenous knowledge on medicinal flora from communities residing near Swat River (Suvastu) and in high mountainous areas in Swat-Pakistan.

AIM OF THE STUDY The present study aimed at documenting the indigenous knowledge on medicinal flora from the local communities residing near Swat River and high mountainous areas in Swat, Pakistan. MATERIALS AND METHODS Field study was carried out in 2013-14. The area was visited 17 times, and semi-structured and open-ended interviews were conducted in both the plain area nearby the Swat River (4 villages) and mountains (7 villages). A total of 83 erudite informants participated in the interviews with their prior consent. The collected data were analysed through quantitative indices viz., medicinal use value (MUV), relative frequency citation (RFC), family use value (FUV), informant consensus factor (ICF) and fidelity level (FL). RESULTS A total of 78 species belonging to 45 vascular plant families were documented as medicinally important. Lamiaceae was observed the dominant medicinal plant family with 11 species, leaf was the most used part, and powder the most preferred preparation type. Maximum MUV (1) was shared by 15 species while maximum RFC (0.385) was demonstrated by both Brassica juncea and Lepidium sativum. Smaller families with mostly 1 species showed higher FUV while among the larger families Amaryllidaceae and Papaveraceae showed maximum FUV of 0.89 and 0.87, respectively. Highest FL was shown by Mentha arvensis (70%) against gastrointestinal diseases. Highest ICF was demonstrated by hepatic disorders (0.93). Furthermore, we revealed 108 indigenous herbal medicinal recipes, 6 plants added to the medicinal plant trade list, 1 species (Vincetoxicum arnottianum) reported for the first time as medicinal plant, and several new uses for 49 well known medicinal plants. CONCLUSION The present survey documents diverse plant species that are utilised by local communities for treating a broad spectrum of disorders. Quantitative indices helped in marking important and most preferred plants. To conclude, we recommend the plants for pharmacological studies, documenting new uses, especially Vincetoxicum arnottianum, never screened before.

Bioinformatics based structural characterization of glucose dehydrogenase (gdh) gene and growth promoting activity of Leclercia sp. QAU-66

Glucose dehydrogenase (GDH; EC 1.1. 5.2) is the member of quinoproteins group that use the redox cofactor pyrroloquinoline quinoine, calcium ions and glucose as substrate for its activity. In present study, Leclercia sp. QAU-66, isolated from rhizosphere of Vigna mungo, was characterized for phosphate solubilization and the role of GDH in plant growth promotion of Phaseolus vulgaris. The strain QAU-66 had ability to solubilize phosphorus and significantly (p ≤ 0.05) promoted the shoot and root lengths of Phaseolus vulgaris. The structural determination of GDH protein was carried out using bioinformatics tools like Pfam, InterProScan, I-TASSER and COFACTOR. These tools predicted the structural based functional homology of pyrroloquinoline quinone domains in GDH. GDH of Leclercia sp. QAU-66 is one of the main factor that involved in plant growth promotion and provides a solid background for further research in plant growth promoting activities.

The diversity of Rhizobia, Sinorhizobia and novel non-Rhizobial Paenibacillus nodulating wild herbaceous legumes

The objective of the present study was to isolate and characterize nodulating bacteria associated with wild legumes. For this purpose, we recovered twenty isolates from root nodules of five wild legume species: Melilotus alles, Melilotus officinalis, Trifolium pratense, Trifolium repens and Medicago sp. Most of the isolates were morphologically analogous with only few exceptions in colony shape, appearance and incubation time. All isolates were Gram negative except T.P2-4. Random amplification of polymorphic DNA showed genetic variation among isolates. The 16S rRNA sequence analysis revealed these isolates as Rhizobium, Sinorhizobium and Paenibacillus. Each of these was also screened for nod D and nod F genes with marked variation at these loci; however, the nucleotide sequence analysis confirmed the presence of nod genes. The assignment of strains to their hosts revealed a unique symbiotic association of Paenibacillus sp. nodulating T .pratense which is being reported here for the first time.

Taxonomic implications of foliar epidermis in Impatiens (Balsaminaceae): Investigating 12 Pakistani taxa as an example

Abstract Impatiens L. is taxonomically a complicated genus that necessitates fresh characterization to resolve its taxonomy. This study therefore aimed at ascertaining micro-morphological characters in the foliar epidermis of Impatiens. For this purpose, we gathered twelve taxa (10 species and 2 subspecies) mostly from the northern regions of Pakistan and studied qualitative and quantitative characters depicting variation in epidermal cells, stomata, and trichomes on both surfaces. Multicellular, uniseriate trichomes were observed only in I. balsamina L., and raphides in I. balfourii Hook. f. and I. thomsonii Hook. f. while stomata were exclusively anomocytic in all species. Morphological variation in the foliar epidermal characters and their diagnostic value has been depicted by constructing a taxonomic key to all taxa. The study revealed that although foliar epidermal characters in Impatiens did not desirably assort taxa on the basis of structural topographies, yet demonstrated adequate variation distinguishing the Pakistani taxa.

Phenotypic and genotypic variability for durable resistance to yellow rust (Puccinia striiformis f. sp. tritici) in Pakistan wheat cultivars

The wheat crop remains vulnerable to all three rust diseases (leaf rust, stem rust and yellow rust) caused by Puccinia spp. according to the prevalence of the pathogen in different wheat-growing areas worldwide. Stripe rust or yellow rust caused by Puccinia striiformis f. sp. tritici is the most significant rust pathogen which prefers cool, moist areas and highlands. The pathogen is recognised as responsible for huge production losses in wheat. Genetic variation in pathogen makes its control difficult. Therefore, resistance against all the races of the pathogen known as durable or race-non-specific resistance is preferred. The present study was carried out to identify durable resistance against stripe rust in selected wheat cultivars from Pakistan through seedling testing, field evaluation at adult stage, morphological marker studies and marker-assisted selection. Results revealed that 4% of the cultivars were resistant at the seedling stage while the rest were susceptible or intermediate. To confirm their field resistance, the same cultivars were evaluated under field conditions at Cereal Crops Research Institute Pirsabak (located in Khyber Pakhtunkhwa, KP) a hot spot of stripe rust in Pakistan. Observations exhibited that at the adult stage 4% of the cultivars were resistant, 70% intermediate or moderately resistant while the others were highly susceptible. Leaf tip necrosis was observed in 30% of the cultivars. Wheat cultivars showing susceptibility at the seedling stage were highly to moderately resistant at adult stage showing durable resistance. For further validation, morphological markers were also observed in cultivars indicating the presence of Yr18/Lr34 gene. Eleven cultivars (C-518, Mexipak, Kohinoor-83, Faisalabad-83, Zardana-93, Shahkar-95, Moomal-2002, Wattan-94, Pasban-90, Kiran-95, and Haider-2000) were identified, having durable or race non-specific resistance against stripe rust. These cultivars can further be utilised in wheat breeding programmes for deploying durable resistance to attain long lasting control against stripe rust.

Melampsora pakistanica sp. nov., a new rust fungus on Euphorbia helioscopia (Sun spurge) from Pakistan

A rust fungus was found on the leaves of Euphorbia helioscopia during a field study in Pakistan. Previously, Melampsora euphorbiae, M. euphorbiae-gerardianae and M. helioscopiae have been reported on E. helioscopia, the first two of which are also known from Pakistan. Morphological observations of the newly collected rust samples detected some differences from the previously described Melampsora species on E. helioscopia. The molecular analysis of the ITS and LSU sequences also detected that the rust is different from the previously reported rusts described from E. helioscopia. Based on both morphological comparisons and sequence analysis, the rust is described here as M. pakistanica sp. nov. This species could have potential as a bio-control agent against its host plant—E. helioscopia—which is a weed of wheat fields in Pakistan and elsewhere in the world.

Evaluation of Glucose Dehydrogenase and Pyrroloquinoline Quinine (pqq) Mutagenesis that Renders Functional Inadequacies in Host Plants.

The rhizospheric zone abutting plant roots usually clutches a wealth of microbes. In the recent past, enormous genetic resources have been excavated with potential applications in host plant interaction and ancillary aspects. Two Pseudomonas strains were isolated and identified through 16S rRNA and rpoD sequence analyses as P. fluorescens QAU67 and P. putida QAU90. Initial biochemical characterization and their root-colonizing traits indicated their potential role in plant growth promotion. Such aerobic systems, involved in gluconic acid production and phosphate solubilization, essentially require the pyrroloquinoline quinine (PQQ)- dependent glucose dehydrogenase (GDH) in the genome. The PCR screening and amplification of GDH and PQQ and subsequent induction of mutagenesis characterized their possible role as antioxidants as well as in growth promotion, as probed in vitro in lettuce and in vivo in rice, bean, and tomato plants. The results showed significant differences (p < or = 0.05) in parameters of plant height, fresh weight, and dry weight, etc., deciphering a clear and in fact complementary role of GDH and PQQ in plant growth promotion. Our study not only provides direct evidence of the in vivo role of GDH and PQQ in host plants but also reveals their functional inadequacy in the event of mutation at either of these loci.

Identification and characterization of rhizospheric microbial diversity by 16S ribosomal RNA gene sequencing

In the present study, samples of rhizosphere and root nodules were collected from different areas of Pakistan to isolate plant growth promoting rhizobacteria. Identification of bacterial isolates was made by 16S rRNA gene sequence analysis and taxonomical confirmation on EzTaxon Server. The identified bacterial strains were belonged to 5 genera i.e. Ensifer, Bacillus, Pseudomona, Leclercia and Rhizobium. Phylogenetic analysis inferred from 16S rRNA gene sequences showed the evolutionary relationship of bacterial strains with the respective genera. Based on phylogenetic analysis, some candidate novel species were also identified. The bacterial strains were also characterized for morphological, physiological, biochemical tests and glucose dehydrogenase (gdh) gene that involved in the phosphate solublization using cofactor pyrroloquinolone quinone (PQQ). Seven rhizoshperic and 3 root nodulating stains are positive for gdh gene. Furthermore, this study confirms a novel association between microbes and their hosts like field grown crops, leguminous and non-leguminous plants. It was concluded that a diverse group of bacterial population exist in the rhizosphere and root nodules that might be useful in evaluating the mechanisms behind plant microbial interactions and strains QAU-63 and QAU-68 have sequence similarity of 97 and 95% which might be declared as novel after further taxonomic characterization.