Characterisation of ganoderma species using morphological, molecular and biochemical markers and evaluation of substrate enhancement influence on their development and biochemical profile = Ukwehlukanisa ngemisebenzi kwezinhlobo zeGanoderma ngokusebenzisa isakhiwo, imolekhyula kanye nezakhi zesayenzi yokuphilayo kanye nokuhlola umthelela wokuthuthuka kwendawo lapho okuphilayo kumila khona ekuthuthukeni kwesimo sesayensi yokuphilayo.

Abstract

Ganoderma, also known as Reishi mushroom, is used for its potential health benefits in several countries. The current study characterised Ganoderma species using molecular and biochemical markers and evaluated the substrate enhancement influence on its development and biochemical profile. The overall research study consisted of four objectives. The first objective focused on the isolation and characterisation of fifteen fungal specimens collected from the three provinces of South Africa, namely, Mpumalanga, KwaZulu-Natal, and North- West. Five fungal specimens were collected in each province and growth media potato dextrose agar (PDA), malt extract agar (MEA), and sabouraud dextrose agar (SDA) were used to grow sample isolates. After 8 days of incubation, MEA recorded the highest mycelial diameter followed by PDA and SDA. Samples were identified using comparative morphology traits and supported by internal transcribed spacer region (ITS) and phylogenetic analyses. Based on the ITS of ribosomal DNA, fungal samples KG3SY219 and MG1SY119 were found to be closely related to Ganoderma resinaceum and Ganoderma austroafricanum, respectively. The species were further characterised by biochemical compounds, including antioxidants, proteins, essential elements, and heavy metals. The antioxidant capacity exhibited a higher radical scavenging activity in G. austroafricanum compared to G. resinaceum. The concentrations of total phenolics, flavonoids, proteins, essential elements and heavy metals were more abundant in G. austroafricanum compared to G. resinaceum. This study also evaluated the effect of different growth conditions on mycelial growth and development of Ganoderma austroafricanum and Ganoderma resinaceum. The experimental treatments included three levels of pH (4, 6 & 8), temperature (20, 25 & 30 °C), and different types of plant residues namely; beech sawdust (BS), sugarcane bagasse (SB), and buffalo grass (BG). Three independent in vitro experiments were conducted, PDA and MEA were used as standard growth media to grow each fungal species. Mycelial growth and development were measured over 9 days where they reached maximum growth. The culture media pH results demonstrated that the maximum growth for mycelia was reached on day 9 for both species. Typically, G. resinaceum showed the highest mycelial growth for both cultures except for days 6 and 9 where the mycelial growth of the species was decreased by low levels (pH 4) and high levels (pH 8), respectively. The addition of BG to the growth media delayed the mycelial growth of G. resinaceum for both growth media (PD+MEA). Experiment three investigated the effect of different substrates on the development, total biomass, and biochemical profile of Ganoderma species. The experiment involved growing G. austroafricanum (GA) and G. resinaceum (GR) on different substrates; beech sawdust (BS), sugarcane bagasse (SB), and buffalo grass (BG), and suspension of all substrates (BSSBBG). The growth parameters such as pileus size, weight, total biomass, and biological efficiency were measured. Additionally, each substrate was analysed for biochemical composition. The biochemical composition of the harvested samples was also analysed to determine the levels of biochemical compounds such as minerals, antioxidants, and protein. The substrate pH levels demonstrated that all substrates were within the optimal growth pH range (5-6). SB exhibited greater levels in the majority of essential elements such as Zn and K, also, heavy metals Pb and Hg. The results on the development and total biomass production of Ganoderma species revealed significant variations across different substrates. In terms of development, GRBS was faster to reach the 100% rate of all production parameters in 40-52 days after inoculation. However, GASB exhibited higher quantities in total yield and biological efficiency. In addition, pileus from GASB demonstrated higher concentrations of all evaluated biochemical compounds. GASB also yielded higher levels of DPPH, phenolic compounds, flavonoids, and protein. Experiment four examined the impact of substrate fortified with essential elements on the development, total biomass, and biochemical compounds of Ganoderma species. The experiment involved growing G. austroafricanum (GA) and G. resinaceum (GR) on beech enhanced with elements; no element (Control), Zn(NO3)2.6H2O (Zn), Fe2SO4 7H2O (Fe), Na2SeO3 (Se), and suspension of all essential elements (ZnFeSe). The growth parameters such as the pileus size, weight, total biomass, and biological efficiency were measured. In addition, the biochemical profile of Ganoderma spp. was analysed to evaluate the concentration of compounds. The development and total biomass production findings for the substrate fortified with essential elements exhibited significant differences. GRZn developed expeditious, reaching 100% of all production parameters in 52 days after inoculation. In comparison to all treatments, GAFe showed larger quantities in total yield and biological efficiency. The substrate enhancement with Zn had a significant increase in the majority of minerals. GAZn exhibited higher concentrations of essential elements such as Zn, K, and Mg. Higher levels of heavy metals such as Cd, Pb, and As were recorded from GAControl. GASe produced higher levels of DPPH, phenolic compounds, flavonoids, and protein reading. These findings demonstrate the variability of morphological characteristics, biochemical compounds, and growth conditions requirements between Ganoderma species. These findings provide valuable insights into the diversity, taxonomy, and potential therapeutic applications of Ganoderma species in South Africa. Further investigation is required to identify Ganoderma species and its pharmaceutical properties.

Iqoqa. Lolu cwaningo lwehlukanisa ngezinhlobo zokusebenza izinhlobo ze-Ganoderma ngokusebenzisa imolekhyula kanye nezakhi zesayensi yokuphilayo futhi luhlole ukuthuthuka kwendawo lapho okuphilayo kumila khona esimweni sakho sokuthuthuka kanye nesayensi yokuphilayo. Ucwaningo selulonke luhlanganise izinhlosongqangi ezine. Inhlosongqangi yokuqala igxile ekwahlukaniseni kanye nasekuhleleni ngezinhlobo zomsebenzi ulibofuzo lokhunta oluyishumi nanhlanu oluqoqwe ezifundazweni ezintathu zaseNingizimu Afrikha, nokuyiMpumalanga, KwaZulu-Natali, kanye nase-North-West. Uhlobo lolibofuzo lokhunta oluyisihlanu lwaqoqwa esifundazweni ngasinye kanye nendlela yokukhula kwe-potato dextrose agar (PDA), i-malt extract agar (MEA), kanye ne-sabouraud dextrose agar (SDA) kwasetshenziswa ukutshala amaqoqo amasampula. Emva kwezinsuku zokukhulisa, i-MEA yaqopha ingxenye enkulu yesikhunta engaphindaphindi nge-PDA kanye ne-SDA. Amasampula atholakala ngokusebenzisa izinkomba zezakhiwo eziqhathanisekayo bese sesekwa ngesahlukaniso sezifunda zangaphakathi ama-internal transcribed spacer (ITS) kanye nokuhlaziya oku-phylogenetic. Ngokwe-ITS of ribosomal DNA, amasampula okhunta i-KG3SY219 kanye ne-MG1SY119 kwatholakala kuhlobene kakhulu kwi-Ganoderma resinaceum kanye ne-Ganoderma austroafricanum, ngokwehlukana kwakho. Lolu cwaningo lwahlola futhi umthelela wezimo zokukhula kwengxenye yesikhunta engaziphindaphindi kanye nokukhula kwe-Ganoderma austroafricanum kanye ne-Ganoderma resinaceum. Ukusetshenzwa kolingo kubalwa amazingeni amathathu e-pH (4, 6 & 8), izinga lokushisa (20, 25 & 30 °C), kanye nezinhlobo ezahlukene zezinsalela zesitshalo okuyi-beech sawdust (BS), i-sugarcane bagasse (SB), kanye ne-buffalo grass (BG). Imizamo emithathu engaphandle ezimele yenziwa, i-PDA kanye ne-MEA kwasetshenziswa njengezindlela zokukhula ezejwayelekile ukukhulisa uhlobo ngalunye lokhunta. Izindlela zokukhula kanye nokuthuthuka kwakalwa ezinsukwini eziyi-9 lapho zafinyelela ekukhuleni kwazo okwanele. Imiphumela yezindlela zokwenza ze-pH yaveza ukuthi ukukhula okukhulu kwengxenye yesikhunta engaziphindaphindi kwafinyeleleka kuyona osukwini lwesi-9 kuzona zombili izinhlobo. Umzamo wesithathu wahlola umthelela wezindawo ezintathu lapho okuphilayo kumila khona ekukhuleni, inani lebhayomasi, kanye nesimo sesayensi yokuphilayo kwezinhlobo ze-Ganoderma. Umzamo wabandakanya ukukhulisa i-G. austroafricanum (GA) kanye ne-G. resinaceum (GR) ezindaweni ezahlukene lapho okuphilayo kumila khona; i-beech sawdust (BS), i-sugarcane bagasse (SB), kanye ne-buffalo grass (BG), kanye nokumiswa kwezindawo lapho okuphilayo kumila khona (BSSBBG). Ukukhula kwemikhawuko efana nobungakho besibalo, isisindo, inani lebhayomasi, kanye nokusebenza kahle kokuphilayo kwakalwa. Okunye futhi, le ndawo lapho okuphilayo kumila khona kwahlaziywa ngenhlanganisela yesayensi yokuphilayo. Inhlanganisela yesayensi yokuphilayo yamasampula avunwa yahlaziyiwa nayo ukuthola amazinga engxubevange yesayensi yokuphilayo efana namaminerali, izakhi ezilwa nobuthi, kanye namaphrotheni. Amazinga endawo lapho okuphilayo kumila khona aveza ukuthi zonke izindawo lapho okuphilayo kumila khona kwakuphakathi nokukhula okulindelekile esigabeni se-pH (5-6). I-SB yaveza amazinga aphezulu ezingxenyeni ezibalulekile eziningi ezifana ne-Zn kanye ne-K, futhi, nensimbi ye-Pb kanye ne-Hg. Imiphumela ekuthuthukeni kanye nasekukhiqizekeni kwenani lebhayomasi lezinhlobo ze-Ganoderma wavela umehluko omkhulu ezindaweni ezahlukene lapho okuphilayo kumila khona. Umzamo wesine wahlola ukuthi umthelela wendawo lapho okuphilayo kumila khona wagqugquzela ngezinhlayiya ezibalulekile ekukhulisekeni, inani lebhayomazi, kanye nezinhlayiya zesayensi yokuphilayo kwezinhlobo ze-Ganoderma. Umzamo wafaka i-G. austroafricanum (GA) ekhulayo kanye ne-G. resinaceum (GR) ohlotsheni lwesihlahla se-beech esinothiswe ngesinhlayiya; ayikho inhlayiya (Ukwengamela), i-Zn(NO3)2.6H2O (Zn), i-Fe2SO4 7H2O (Fe), i-Na2SeO3 (Se), kanye nokumiswa kwazo zonke izinhlayiya (ZnFeSe). Ukukhula kwemikhawuko efana nobungako besivalo, isisindo, inani lebhayomazi, kanye nokusebenza kokuphilayo kwakalwa. Okunye futhi, isimo sesayensi yokuphilayo se-Ganoderma spp. sahlaziywa ukuhlola ukugxila kwezinxayiya. I-GRZn yathuthuka ngokushesha, yafika ku-100% wemikhawuko yonke yomkhiqizo ezinsukwini ezingama-52 emva kokujovwa. Uma kuqhathaniswa nakho konke ukusetshenzwa, i-GAFe yaveza izinombolo ezinkulu zamanani emiphumela kanye nokusebenza kahle kokuphilayo. Ukukhuliswa kwendawo lapho okuphilayo kumila khona ngo-Zn kwaba nokuphakama okukhulu eningini lamaminerali. I-GAZn yaveza ukugxila okuphakeme kwezinhlayiya ezibalulekile ezifana ne-Zn, i-K, kanye ne-Mg, amazinga aphezulu ensimbi esindayo efana ne-Cd, i-Pb, kanye ne-As kwaqoshwa nge-GAControl. I-GASe yakhiqiza amazinga aphezulu e-DPPH, izinhlayiya ezi-phenolic, ama-flavonoids, kanye nokuthola iphrotheni. Le miphumela yaveza ukwehlukahluka kwezakhiwo zokwehlukanisa ngokomsebenzi, izinhlayiya zesayensi yokuphilayo, kanye nokukhula kwezidingo zesimo phakathi kwezinhlobo ze-Ganoderma. Le miphumela inikeza imibono ebalulekile ekwehlukeni, ukuqoqela ndawonye, kanye nokusetshenziswa kokwelapha okungenzeka kwezinhlobo ze-Ganoderma eNingizimu Afrikha. Olunye uphenyo luyadingeka ukuthola izinhlobo ze-Ganoderma kanye nezinhlaka zayo zokwelapha.