Impact of Ionic and Heavy Metal Stress on Secondary Metabolite Accumulation in Ajuga bracteosa Calli | InformativeBD

Roby Gul, from the institute of Pakistan. Gul-Lalay, from the institute of Pakistan. Safdar Hussain Shah, from the institute of Pakistan. Saad Hussain Shah, from the institute of Pakistan. And Fazal Umer, from the institute of Pakistan. wrote a Research Article about, Impact of Ionic and Heavy Metal Stress on Secondary Metabolite Accumulation in Ajuga bracteosa Calli. Entitled, Effects of ionic and heavy metal stress on secondary metabolites accumulation in calli of Ajuga Bracteosa L.. This research paper published by the International Journal of Biosciences(IJB). an open access scholarly research journal on Biosciences. under the affiliation of the International Network For Natural Sciences| INNSpub. an open access multidisciplinary research journal publisher.

Abstract

Ajuga bracteosa L, a medicinal plant that contains numerous phytochemicals, is becoming endangered worldwide due to the lack of knowledge and overexploitation. Research efforts are necessary for its conservation, preservation, and secondary metabolite production. In this study, an attempt was made to develop calli from this plant and evaluate the effects of abiotic stresses, i.e., NaCl and Cd(NO3)2, on the relative growth rate, total phenolic contents (TPC), proline contents, sugar contents, total flavonoid contents (TFC), alpha-amino acids, and antioxidant activity of A. bracteosa at the cellular (calli) level. NaCl and Cd (NO3)2 reduced the biomass production in terms of relative growth rate (RGR) but enhanced metabolite accumulation. The highest total phenolic contents (40µg/g), proline contents (0.9µg/g), total sugar contents (90mM/g), and total flavonoid contents (5mg/g) were recorded at 150mM NaCl, while the maximum total antioxidant activity (900µM/g) was measured at 50mM NaCl. On the other hand, the lowest amino acid contents (5µM), phenolic contents (27µg), and proline contents (0.5µg) were found at 50mM NaCl. Similarly, the highest level of Cd(NO3)2 stress (8mM) resulted in insignificantly increased production of phenolics (22µg/g), proline (0.9µg/g), total sugars (92mM), alpha-amino acids (7µM), and total antioxidant activity (850µM). This study not only provides an ex-situ conservation strategy for a highly exploited medicinal herb but can also be used as a commercial production methodology for valuable secondary metabolites.

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Introduction

All plants either cultivated or wild ones having active ingredients, which are used for therapeutic purpose, are called medicinal plants. The classification of plants into medicinal plants, poisonous plants, arable plants, ornamental, weeds etc. is always based on a human element and reflect a certain attitude, a purpose or a goal or economic interests (Imre, 2012). In broad sense, any plant is considered as medicinal herb, including fruits, vegetables, and arable plants. Humans depend on plants for various purposes like, food, medicines and shelter (Morgan et al., 1981). Now days due to overexploitation, overgrazing and lack of knowledge lot of plants species becomes endangered (Sher et al., 2008). Round about 10% flora and 709 plants in Pakistan are endangered (Shinwari et al., 2010), among these 580 are flowering plants (Khan et al., 2011). From medicinal plants approximately 25% of modern medicines are directly or indirectly derived. Distributions of these plants are worldwide but they are mostly abundant in tropical regions. These plants are abundant in Himalaya and upper Gangetic plans (Singh et al., 2006; Israili et al., 2009).

Ajuga bracteosa L, a medicinal plant is locally called Butey, in English “Bungle”, in Sansikrit “Nilkanthi” and in Kashmir “Jan-I adam”. In Northern areas of Pakistan due to its sour taste, it is called kauri booti (Jan et al., 2014). A. bracteosa contains numerous phytochemicals such as flavonol glycosides, iridiod, glycosides, neo-clerodane diterpenoids, phytoecdysones and ergosterol-5, 8-endoperoxide (Castro, 2011; Kayani et al., 2016). From A.bracteosa wall, phenolic compound Ajuganane which is a new compound along with three other compound 7- hydroxy-3,6,3’,4’-tetramethoxyflavone, 3,4’- dihydroxy-3,6,7-trimethoxyflavone and urasolic were isolated (Hussain et al., 2012). Mostly A. bracteosa are used for treatment of hepatitis in Taiwan (Hsieh et al., 2011). It is used for the treatment of gout, palsy, amenorrhea and rheumatism (Kaithwas et al., 2012). For treating malaria, it is also used in many countries (Chandal and Bagai, 2011). A. bracteosa is also used for treatment of different diseases like, neuro inflammatory diseases, pneumonia, gout and hepatitis in Asian countries (Nisar et al., 2014).

In India the plant leaves, flowers and barks are used for treatment of diabetes, cancer and malaria etc. (Pal et al., 2014). Leaves extracts are used for headache, throats and ear infection, acne, pimples and also used as blood purifier etc. (Bisht et al., 2013). Plant extract is also used against gastric ulcer, protozoa infection, liver fibrosis, hypoglycemia, anti-inflammatory, anticancer, anti-arthritic, anti-plasmodial, immunoregulatory and insecticidal. The root juice of plant is used for treatment of dysentery and diarrhea; leaves are used as a substituent for quinine for treatment of fever (Pal and Pawar, 2011).

Similarly, different medicinal activity of A. bracteosa like, antibacterial, astringent and anthelmintic is considered in folk medicine. In KPK, an ethno botanic survey of 92 medicinal plants was done, in which A. bracteosa, due to its high medicinal value (antijaundice, antiasthma, ant cough and cooling agent) listed at the top (Ibrar et al., 2015). Studies investigated the anticoagulant, anti-depressant, antioxidant an inflammatory activities of A. bracteosa. A. bracteosa adventitious root also contains Gallic acid, Rutin, Catechin, Myricetin and apigenin (Ali et al., 2018). Due to over exploitation of medicinal plants from last decades’ important plants are at the risk of extinction. A.bracteosa has rated as an endangered flowering medicinal plant species belongs to family L miaceae. It contains 170 genera of medicinal herb, and is mostly found in temperate regions (Hafeez et al., 2017).

Plant tissue culture an important component of biotechnology, generally used for micro propagation, production of virus free plants, rapid multiplication of clones, suspension culture, embryo culture and organ culture etc. One of the most important uses of this technology is to induce and multiply callus culture for extraction of secondary metabolites (active ingredients) (Hussain et al., 2011). Abiotic stresses like, salt, temperature, light, drought and heavy metals etc. affect secondary metabolites production like phenol, proline, flavonoid and amino acids etc. (Akula and Ravishankar, 2011). However, in some plant species heavy metals alter the nature of secondary metabolites (Nasim and Dhir, 2010).

The current study was designed with the following objectives:

1. To induced and multiply callus culture of A. bracteosa.

 2. To determine the antioxidant activity and secondary metabolites i-e phenols, flavonoids, proline, amino acids, total sugar contents in response to ionic and heavy metal stress (NaCl and Cd(NO3)2).

Reference

Akula R, Ravishankar GA. 2011. Influence of abiotic stress signals on secondarymetabolites in plants. Plant signaling and behavior 6(11):1720-1731.

Ali H, Khan MA, Kayani WK, Khan T, Khan RS. 2018. Thidiazuron regulated growth, secondary metabolism and essential oil profiles in shoot cultures of Ajuga bracteosa. Industrial crops andproducts 121, 418-427.

Balestrasse KB, Benavides MP, Gallego SM, and Tomaro ML 2003. Effect of cadmium stress on nitrogen metabolism in nodules and roots of soybean plants. Functional Plant Biology 30(1), 57-64.

Bandhe-hagh A, Toorchi M, Mohammadi A, Chaparzadeh N, Hosseini Salekdeh G. Kazemnia H. 2008. Growth and osmotic djustment of canola genotype in reponse to salinity. Journal of Food Agriculture Environment 6(2), 201-208.

Bisht VK, Negi JS, Bhohn AK, Sundriyal RC (2013).Traditional use of medicinal plants in district Chamoli, Uttarakhand, India. Journal of Medicinal Plants Research7(15), 918-929.

Castro A, Coll J, Arfan M. 2011. neo-Clerodane diterpenoids from Ajuga bracteosa. Journal of natural products 74(5), 1036-1041.

Chandel S, Bagai N. 2011. Screening of antiplasmodial efficacy of Ajuga bracteosa Wall ex. Benth. Parasitology research 108(4), 801-805.

El-Esawi MA, Elansary HO, El-Shanhorey NA, Abdel-Hamid AM, Ali HM, Elshikh MS. 2017. Salicylic acid-regulated antioxidant mechanisms and gene expression enhance rosemary performance under saline conditions. Frontiers in Physiology 8, 716.

García‐Caparrós P, Llanderal A, Pestana M, Correia PJ, Lao MT. 2017. Lavandula multifida response to salinity: Growth, nutrient uptake, and physiological changes. Journal of Plant Nutrition and Soil Science 180(1), 96-104.

Hafeez K, Andleeb S, Ghousa TG, Mustafa R, Naseer A, Shafique I, Akhter K. 2017. Phytochemical screening, alpha-glucosidase inhibition, antibacterial and antioxidant potential of Ajuga bracteosa extracts. Current pharmaceutical biotechnology 18(4), 336-342.

Hare P, Cress WA, Van Staden J. 1998. Dissecting the roles of osmolyte accumulation during stress. Plant. Cell. Environ 21, 535-553.

Hong Z, Lakkinel K, Zhang Z, Verma DPS. 2002. Removel of feedback inhibition of pyrroline-5-carboylase sythetase results in increased proline accumulation and protecion of plants from osmotic stress. Plant Physiol 122, 1129-11136.

Hsieh WT, Liu YT, Lin WC 2011. Anti-inflammatory properties of Ajuga bracteosa in vivo and in vitro study and their effects on mouse model of liver fibrosis. Journal of ethnopharmacology 135(1), 116-125.

Hussain A, Qarshi IA, Nazir H, Ullah I. 2012. Plant tissue culture: current status and opportunities. Recent advances in plant in vitro culture 1-28.

Hussain J, Begum N, Hussain H, Khan FU, Rehman NU, Al-Harrasi A, Ali L. 2012. Ajuganane: a new phenolic compound from Ajuga bracteosa. Natural product communications 7(5), 1934578 X 1200700518.

Ibrahim MH, Chee Kong Y, Mohd Zain NA. 2017. Effect of cadmium and copper exposure on growth, secondary metabolites and antioxidant activity in the medicinal plant Sambung Nyawa (Gynura procumbens (Lour.) Merr). Molecules 22(10), 1623.

Ibrar M, Rauf A, Hadda TB, Mubarak MS, Patel S. 2015. Quantitative ethnobotanical survey of medicinal flora thriving in Malakand Pass Hills, Khyber Pakhtunkhwa. Pakistan. Journal of ethnopharmacology 169, 335-346.

Imre SP, Kilmartin PA, Rutan T, Mauk JL, Nicolau L. 2012. Influence of soil geochemistry on the chemical and aroma profiles of pinot noir wines. Journal of Food, Agriculture and Environment 10(2 Part 1), 280-288.

Israili ZH, Lyoussi B. 2009. Ethnopharmacology of the plants of genus Ajuga. Pakistan Journal of Pharmaceutical Sciences 22(4).

Jan M, Singh S, Kaloo ZA, Maqbool F. 2014. Medicinal importance of Ajuga bracteosa Wall ex Benth.-a review. International Journal of Advanced Research 2(1), 389-94.

Kaithwas G, Gautam R, Jachak SM, Saklani A 2012. Antiarthritic effects of Ajuga bracteosa Wall ex Benth. in acute and chronic models of arthritis in albino rats. Asian Pacific journal of tropical biomedicine 2(3), 185-188.

Kayani WK, Dilshad E, Ahmed T, Ismail H, Mirza B. 2016.Evaluation of Ajuga bracteosa for antioxidant, anti-inflammatory, analgesic, antidepressant and anticoagulant activities. BMC complementary and alternative medicine 16(1), 1-13.

Khan M, ZK Shinwari, Musharaf S. 2011. Conservation and ecological characteristic of Trees in Tehsil Karak Pakistan. Journal of Biodiversity and Environmental Sciences 1(6), 155-164.

Kuntal S, Mankand AU, Redd MN. 2017. Cadmium accumulation and its effects on growth and biochemical parameters in Tagetes erecta L.Journal of Pharmacognosy and Phytochemistry 6(3), 111-115.

Mamdouh M, Mahmoud Y, Omar E,Zeinab B. 2002. Kinetin regulation of growth and secondary metabolism in waterlogging and salinity treated Vigna sinesis and Zea mays. Acta PhysiologiaePlantarum 24(1), 19-27.

Manquián-Cerda K, Escudey M, Zúñiga G, Arancibia-Miranda N, Molina M, Cruces E. 2016. Effect of cadmium on phenolic compounds, antioxidant enzyme activity and oxidative stress in blueberry (Vaccinium corymbosum L.) plantlets grown in vitro. Ecotoxicology and environmental safety 133, 316-326.

Morgan W. 1981. Ethnobotany of the Turkana: use of plants by a pastoral people and their livestock in Kenya. Economic Botany 35(1), 96-130.

Murashige T. 1974. Annual review of plant physiology.Plant propagation through tissue cultures 25(1), 135-166.

Nasim SA, Dhir B. 2010. Heavy metals alter the potency of medicinal plants, in Reviews of environmental contamination and toxicology. Springer 139-149.

Nazir S, Jan H, Tungmunnithum D, Drouet S, Zia M, Hano C, Abbasi BH. 2020. Callus culture of Thai basil is an effective biological system for the production of antioxidants. Molecules 25(20), 4859.

Nisar A, Akhtar N, Hassan A, Banday T, Wani B, Zargar MA. 2014. Effect of Ajuga bracteosa on systemic T-cell immunity in Balb/C mice: dual Th1/Th2 immunostimulatory effects. The American journal of Chinese medicine 42(02), 375-392.

Paek KY, Chandler SF, Thorpe TA. 1988. Physiological effects of Na2SO4 and NaCl on callus cultures of Brassica campestris (Chinese cabbage). Physiologia Plantarum 72(1), 160-166.

Pal A, Pawar R. 2011. A study on Ajuga bracteosa wall ex. Benth for analgesic activity. International Journal of Current Biological  and Medical Science 1(2), 12-14.

Pal A, Toppo FA, Chaurasiya PK, Singour PK, Pawar RS. 2014. In-vitro cytotoxicity study of methanolic fraction from Ajuga Bracteosa wall ex. benth on MCF-7 breast adenocarcinoma and hep-2 larynx carcinoma cell lines. Pharmacognosy research 6(1), 87.

Palma JM, Sandalio LM, Corpas FJ, Romero-Puertas MC, McCarthy I, Luis A. 2002. Plant proteases, protein degradation, and oxidative stress: role of peroxisomes. Plant Physiology and Biochemistry 40(6-8), 521-530.

Parlak KU, Yilmaz DD. 2012. Response of antioxidant defences to Zn stress in three duckweed species. Ecotoxicology and Environmental Safety 85, 52-58.

Redillas MC, Park SH, Lee JW, Kim YS, Jeong JS, Jung H, Kim JK. 2012. Accumulation of trehalose increases soluble sugar contents in rice plants conferring tolerance to drought and salt stress. Plant Biotechnology Reports 6(1), 89-96.

Riadh K, Wided M, Ahmed D, Hanen F, Claude G, Chedly A. 2007. Salinity effects on polyphenol content and antioxident activities in leaves of the halophyte Cakile maritime. Plant Phsiology and Biochemistry 45(3-4), 244-249.

Shah S, Wainwright S, Merrett M. 1990. The interaction of sodium and calcium chlorides and light on growth, potassium nutrition, and proline accumulation in calluscultures of Medicago sativa L. New phytologist 116(1), 37-45.

Sher H, Khan ZD, Khan AU, Hussain F. 2005. Insitu conservation of some selected medicinal plants of Upper Swat. Pakistan. J. Acta Botanica Yunnanica 27, 27-36.

Shinwari ZK. 2010. Medicinal plants research in Pakistan. Journal of medicinal plants research 4(3), 161-176.

Singh N, Mahmood U, Kaul VK, Jirovetz L 2006. A new phthalic acid ester from Ajuga bracteosa. Natural product research 20(06), 593-597.

Usman H, Ullah MA, Jan H, Siddiquah A, Drouet S, Anjum S, Abbasi BH. 2020. Interactive Effects of wide-spectrum monochromatic lights on phytochemical production, antioxidant and biological activities of Solanum xanthocarpum callus cultures. Molecules 25(9), 2201.

Van den Berg R, Haenen GR, van den Berg H, Bast AALT. 1999. Applicability of an improved Trolox equivalent antioxidant capacity (TEAC) assay for evaluation of antioxidant capacity measurements of mixtures. Food Chemistry 66(4), 511-517.

Zahra A, Seyed MG, Ali AG. 2019. Cadmium accumulation and its effects on physiological and biochemical characters of summer savory (Satureja hortensis L.). International journal of phytoremediation 21(12), 1241-1253.

Zheng W, Wang SY. 2001. Antioxident activity and phenolic cmpounds in selected herbs. Journal of Agriculture Food Chemistry 49, 5165-5170.

Zhou Y, Tang N, Huang L, Zhao Y, Tang X, Wang K. 2018. Effects of Salt Stress on Plant Growth, Antioxidant Capacity, Glandular Trichome Density, and Volatile Exudates of Schizonepeta tenuifolia Briq. International journal of molecular sciences 19(1), 252.

Source : Effects of ionic and heavy metal stress on secondary metabolites accumulation in calli of Ajuga Bracteosa L.