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

Khan, A. L.,Gilani, S. A.,Waqas, M.,Al-Hosni, K.,Al-Khiziri, S.,Kim, Y. H.,Ali, L.,Kang, S. M.,Asaf, S.,Shahzad, R. Springer Science + Business Media 2017 Journal of Zhejiang University. Science B. Biomedi Vol.18 No.2

<P>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) mu mol/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.</P>

Elemental allelopathy and antifungal activities of Inula falconeri from Himalaya Pakistan

Khan, A. L.,Hussain, J.,Hamayun, M.,Gilani, S. A.,Kim, Y. H.,Rehman, S. u.,Watanabe, K.,Lee, I. J. Taylor Francis 2010 Acta agriculturæ Scandinavica. Section B, So Vol.60 No.6

First draft genome sequencing of indole acetic acid producing and plant growth promoting fungus Preussia sp. BSL10

Khan, A.L.,Asaf, S.,Khan, A.R.,Al-Harrasi, A.,Al-Rawahi, A.,Lee, I.J. Elsevier Science Publishers 2016 Journal of biotechnology Vol.225 No.-

<P>Preussia sp. BSL10, family Sporormiaceae, was actively producing phytohormone (indole-3-acetic acid) and extra-cellular enzymes (phosphatases and glucosidases). The fungus was also promoting the growth of arid-land tree-Boswellia sacra. Looking at such prospects of this fungus, we sequenced its draft genome for the first time. The Illumina based sequence analysis reveals an approximate genome size of 31.4 Mbp for Preussia sp. BSL10. Based on ab initio gene prediction, total 32,312 coding sequences were annotated consisting of 11,967 coding genes, pseudogenes, and 221 tRNA genes. Furthermore, 321 carbohydrate-active enzymes were predicted and classified into many functional families. (C) 2016 Elsevier B.V. All rights reserved.</P>

Tpl-2 kinase downregulates the activity of p53 and enhances signaling pathways leading to activation of activator protein 1 induced by EGF

Khanal, P.,Lee, K.-Y.,Kang, K.-W.,Kang, B. S.,Choi, H. S. Oxford University Press 2009 Carcinogenesis Vol.30 No.4

<P>Tumor progression locus-2 (Tpl-2) kinase is a member of the mitogen-activated protein kinase kinase kinase family that has been implicated in cellular transformation. The enhanced expression of this protein has been shown to activate both the mitogen-activated protein kinase and c-Jun N-terminal kinase pathways. However, the molecular mechanisms responsible for the oncogenic potential of Tpl-2 are still largely unknown. Here, we showed that Tpl-2 interacted with p53 both in vitro and ex vivo. The overexpression of Tpl-2 inhibited the epidermal growth factor (EGF)-induced p53 phosphorylation (Ser15) through upregulating the activity of protein phosphatase 2A, which interacted with p53 stimulated by EGF. Also, the EGF-induced p53 activity was suppressed in the Tpl-2 wild-type (WT)-transfected HEK 293 cells, but had no effect in the Tpl-2-mutant (S413A)-transfected cells. Furthermore, introduction of small interfering RNA-Tpl-2 into HEK 293 cells resulted in decreased cell viability compared with only adenovirus-p53-infected cells. In addition, the Tpl-2 WT, but not Tpl-2 mutant (S413A), showed increased EGF-induced c-fos promoter activity, followed by activator protein 1 (AP-1) transactivation activity, which was associated with the cell transformation prompted by the H-Ras-Tpl-2-AP-1 signaling axis. These results indicated that the Ser413 of Tpl-2 plays an important role in EGF-induced carcinogenesis as well as inactivation of the p53.</P>

Status of Milk Fat Conjugated Linoleic Acid (CLA) in Selected Commercial Dairies

Khanal, R.C.,Dhiman, T.R. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.10

Because of the increasing evidence of potential benefits of conjugated linoleic acid (CLA) on human health, there is a need to investigate its status in commercial dairies and develop feeding strategies to enhance the content and supply of CLA in milk and milk products. A two-year experiment was conducted to study the status of milk fat c-9, t-11 CLA on four selected commercial dairy farms in Utah (two) and Idaho (two), USA. Farms A and C grazed cows on pasture and supplemented with 7.0 kg/cow per day of their respective grain mixes during summer, while conserved forage and grain mix was fed during winter. Farm B fed a total mixed diet all year, with 10% of diet dry matter as fresh cut pasture during summer. Farm D had 1/3 of its cows grazed on pasture and supplemented with a total mixed diet during summer, while the rest were fed a total mixed diet. All cows in Farm D were fed a total mixed diet during winter. Farms A, B, C, and D had on average 80, 400, 150, and 500 milking cows, respectively, with Holstein or its crosses as the major breed. On a year-round basis, Farms A and C produced milk with 60% or more milk fat c-9, t-11 CLA and transvaccenic acid (TVA) contents than Farm B. Similarly, Farm D produced 30% or more c-9, t-11 CLA and TVA in milk than Farm B. Milk fat content of CLA and TVA was 150-200% more during summer compared with winter. Individual cows varied from 0.16 to 2.22% in milk fat c-9, t-11 CLA contents and 89% of the cows had c-9, t-11 CLA contents between 0.3 and 1.0% of milk fat. Individual cow variation was larger on Farms A and C compared with Farm D, with least variation on Farm B. Variation was larger in summer than in winter. The bulk tank milk c-9, t-11 CLA content varied from 0.27 to 1.35% of milk fat. Cows on Farms A and C produced similar or higher amounts of milk fat c-9, t-11 CLA on a daily basis even though their milk yield was lowest among the dairies. Concentration and supplies of c-9, t-11 CLA and TVA were highest from June through September and lowest from February through April, which should be the months for targeting improvement in the content and supply of milk fat c-9, t-11 CLA and TVA.

Proyl isomerase Pin1 facilitates ubiquitin-mediated degradation of cyclin-dependent kinase 10 to induce tamoxifen resistance in breast cancer cells

Khanal, P,Yun, H J,Lim, S C,Ahn, S G,Yoon, H E,Kang, K W,Hong, R,Choi, H S Macmillan Publishers Limited 2012 Oncogene Vol.31 No.34

Endocrine therapies that inhibit estrogen receptor (ER)-α signaling are the most common and effective treatment for ER-α-positive breast cancer. However, the use of these agents is limited by the frequent development of resistance. The aim of this study was to elucidate the mechanisms by which downregulation of CDK10 expression confers resistance to tamoxifen in breast cancer. Here, we show that peptidyl-prolyl isomerase Pin1 downregulates CDK10 protein as a result of its interaction with and ubiquitination of CDK10, thereby affecting CDK10-dependent Raf-1 phosphorylation (S338). Pin1<SUP>–/–</SUP> mouse embryonic fibroblasts (MEFs) show higher CDK10 expression than Pin1<SUP>+/+</SUP> MEFs, whereas CDK10 protein was downregulated in the rescued Pin1<SUP>–/–</SUP> MEFs after reexpression of Pin1. Pin1 silencing in SKBR-3 and MCF7 cells increased the CDK10 expression. In human tamoxifen-resistant breast cancer and tamoxifen-resistant MCF7 cells, immunohistochemical staining and immunoblotting analysis shows an inverse correlation between the expression of CDK10 and the degree of tamoxifen resistance. There was also a positive correlation between the high level of P-Raf-1 (Ser338) and Pin1 in human tamoxifen-resistant breast cancer and tamoxifen-resistant MCF7 (TAMR-MCF7) cells. Importantly, 4-OH tamoxifen (4-OHT), when used in combination with overexpressed CDK10 or Raf-1 inhibitor, increased cleaved PARP and DNA fragmentation to inhibit cologenic growth of MCF7 cells and Tamoxifen-resistant MCF7 cells, respectively. On the basis of these findings, we suggest that the Pin1-mediated CDK10 ubiquitination is a major regulator of tamoxifen-resistant breast cancer cell growth and survival.

α-Glucosidase inhibitors from Duranta repens modulate p53 signaling pathway in diabetes mellitus

Khanal Pukar,Patil B. M. 경희대학교 융합한의과학연구소 2020 Oriental Pharmacy and Experimental Medicine Vol.20 No.3

Hydroalcoholic extract/fraction(s) and scutellarein, a previously reported phytoconstituent from Duranta repens were evaluated for their α-glucosidase inhibitory activity using in vitro method. The reported phytoconstituents were also screened as α-glucosidase inhibitor, probable cytotoxicity, ADMET profile, and side effects via in silico models. Further, probable gene expression profile and pathways involved in diabetes/diabetes complications were also screened. Similarly, network was constructed among α-glucosidase inhibitors, modulated proteins and respective pathways and docking study was performed using autodock4.0. Fraction rich in flavonoids was found to possess the highest α-glucosidase inhibitory activity and the mode of inhibition was uncompetitive. The predicted α-glucosidase inhibitors were less cytotoxic to normal cells with lower side effects compared to acarbose. Similarly, gene-set enrichment analysis identified p53 signaling pathway to be primarily modulated by majority of phytoconstituents. Further, docking study revealed scutellarein to have highest binding affinity with α-glucosidase enzyme. In conclusion, present study identified the fraction rich in flavonoids to possess highest α-glucosidase inhibitory activity and could modulate p53 signaling pathway in diabetic pathogenesis.

Genome editing interventions to combat rice blast disease

Khanale Vaishali,Bhattacharya Anjanabha,Prashar Mohinder,Char Bharat 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.1

Rice is the major food crop of the world. Production of rice is threatened by a number of biotic and abiotic stresses and these could result in a significant yield reduction, sometimes up to 60–100%. In addressing losses to emerging diseases, development of resistance to blast, sheath blight, and false smut is the need of the hour. The traditional approach of using resistance R genes for building sustainable resistance is not viable anymore due to continuously evolving strain of pathogens, thus causing intransigence erosion. The contemporary intervention strategy of identifying and precisely mutating susceptibility genes opens up new vistas for exploiting susceptible genes. Approaches like mutation breeding and transgenic held good promise to further yield increases in rice and provide protection against disease induced yield losses. However, all these approaches either proved inadequate over time or faced regulatory hurdles and were successful to a limited point. Recently, a new genome editing (GE) approach has offered a promising alternative to develop unique alleles in a shorter time frame without linkage drag. Thus, the present review highlights various R, non-R (S) gene choices amenable for genome editing, compared with available conventional breeding approaches, like wide-hybridisation using wild relatives, including new genetic resources. Hence, a range of breeding resources can be employed by crop breeders to assemble disease resistance against emerging diseases of rice. Many successful examples on disease resistance are collated from recently published work. It is hoped that creating new allelic series of variants would be a practical and sustainable solution to address the challenges posed to food security.

Smart Farm Chicken Counting using CCTrans

Khanal Ridip(카날 리딥),Choi Yoochan(최유찬),Lee Joonwhoan(이준환) 한국통신학회 2023 한국통신학회 인공지능 학술대회 논문집 Vol.2023 No.9

Effect of Feeding Urea Treated Rice and Wheat Straw on Intake and Milk Yield of Lactating Buffaloes under Farmers Conditions

Khanal, R.C.,Gurung, D.B.,Kadariya, R.K. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.8

Two experiments were conducted to study the effect of urea treatment of rice and wheat straw on feed intake, dry matter (DM) digestibility and milk yield of lactation buffaloes in their late lactation under farmers' management conditions in the western hills of Nepal during 1995 and 1997. Dry matter intake (DMI) from urea treated rice and wheat straw was not improved significantly (p<0.05) nor the total DMI of the lactating buffaloes was improved significantly. However, feeding urea treated rice straw increased straw DMI by 14.2% and total DMI by 10.63% units over the untreated rice straw. Similarly, the increase in straw and total DMI were 20.18 and 17.40% units over the untreated wheat straw fed animals. Although there was no significant effect of urea treatment of both straw on DM digestibility, it was higher for treated than untreated straw at all locations. An overall increment of 18.1% units for rice straw and 13.3% units for wheat straw was observed. There was a significant effect (p<0.01) of feeding urea treated rice and wheat straw on the milk yield of lactating buffaloes during late lactation under farmers conditions. Post experiment milk yield was also significantly (p<0.05) higher for the animals fed treated straw in both the experiments. Buffalo milk yield was also significantly affected by breed (p<0.01), location (p<0.01) and parity (p<0.01) of the animals. General response of the farmers about the technology and their observed effect on animal performance was also very positive.