Identification and inoculation of fungal strains from Cedrus deodara rhizosphere involve in growth and alleviation of high nitrogen stress

Abstract

Cedrus deodara is economically and ethnobotanically an important forest tree and is shown to be at decline in Northern areas of Pakistan in recent years mainly due to high concentration of Nitrogen in forests. Ectomycorrhizal (ECM) association forming fungi enables the forest trees to develop optimally by absorbing water from the rhizosphere through their absorptive hyphae and by making available the nutrients by mobilization of N and P from the organic substrates. This study was conducted to identify the ECM strains from C. deodara rhizosphere and to analyse the impact of high N load on the C. deodara seedlings to establish N critical load value for coniferous forests of Pakistan. Six new fungal strains were identified from the rhizosphere of C. deodara and were registered at GenBank (NCBI) as Emmia latemarginata strain ACE1, Aspergillus terreus strain ACE2, Purpureocillium lilacinum strain ACE3, Talaromyces pinophilus strain ACE4, A. fumigatus strain ACE5 and T. pinophilus strain ACE6 with accession numbers MH145426, MH145427, MH145428, MH145429, MH145430 and MH547115. Four out of six isolated strains were inoculated with seedlings of C. deodara singly and in consortium (CN) in combination with nitrogen load of 0 (C), 25 (T1), 50 (T2), 100 kg N ha−1 yr−1 (T3). Agronomic, physiological and gene expression studies for ExpansinA4 (EXPA4) and Cystatins (Cys) were made to analyse the impact of fungal strains in relation to high N stress. This study suggests a positive impact of T1 (25 kg N ha−1 yr−1) Nitrogen load and a negative impact of T3 (100 kg N ha−1 yr−1) on growth parameters and expression patterns of EXPA4 and Cys genes. Peroxidase (POX) activity decreased in the order ACE5 > ACE2 > C > ACE3 > ACE1 > CN. However, the results of Superoxide dismutase (SOD) showed decreasing trend in the order ACE5 > C > CN > ACE1 > ACE2 > ACE3. Strain ACE3 was shown to have a positive impact on the seedlings in terms of growth, physiology and expression of genes. Present study suggests that newly identified fungal strains showing positive impact on the growth and physiology of C. deodara could be used for the propagation of this economically important plant in Pakistan after pathogenicity test.

Molecular & Morphological Identification of Entomopathogenic Fungus Purpureocillium lilacinum Isolated From Soil, Balakot Tehsil, Khaber Pakhtunkhwa, Pakistan

“…As a result, six new fungal strains were identi ed ACE4, ACE2, ACE3, ACE5, ACE1 and ACE6 of the Talaromyces pinophilus, Aspergillus terreus, Aspergillus fumigatus, Purpureocillium lilacinum, Emmia latemar ginata, and T. pinophilus strains, respectively. Both areas reported P. lilacinum since the atmospheric conditions in this region are similar to those in the current research area, including temperature and sampling technique (Jamil et al, 2020), and this is the rst report on P. lilacinum from the nothren area of Pakistan. Othere province, such as Pujab, also reported this in the past.…”

“…The whole nucleic acid extraction procedure utilized the CTAB method, with optimizations made to the existing protocol (Jamil, 2020). To extract DNA, the fungal sample was manually ground in a sterile grinder within a 1.5-milliliters microtube.…”

Detection of hepatitis through proteases and protease inhibitors genes expression and identification of HCV untypable genotype in Abbottabad, Pakistan

“…The amplified PCR product was visualized on a 2% agarose gel and 50 bp DNA ladder (Invitrogen, Corp, USA) according to the protocol described by Jamil et al (2020). Phylogenetic analysis.…”