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Disszertáció DOKTORI (PhD) ÉRTEKEZÉS HORVÁTH NÁNDOR MOSONMAGYARÓVÁR 2020 SZÉCHENYI ISTVÁN EGYETEM MEZŐGAZDASÁG- ÉS ÉLELMISZERTUDOMÁNYI KAR MOSONMAGYARÓVÁR Növénytudományi Tanszék Wittmann Antal Növény-, Állat- és Élelmiszer-tudományi Multidiszciplináris Doktori Iskola Doktori Iskola vezetője: Prof. Dr. Ördög Vince egyetemi tanár, az MTA doktora Készült a „Haberlandt Gottlieb Növénytudományi Doktori Program” keretében Programvezető: Prof. Dr. Ördög Vince egyetemi tanár, az MTA doktora Témavezetők: Prof. Dr. Ördög Vince egyetemi tanár, az MTA doktora Dr. habil. Molnár Zoltán, PhD, egyetemi docens A MOSONMAGYARÓVÁRI ALGAGYŰJTEMÉNY (MACC) KORÁBBAN ALAKTANI SZEMPONTBÓL ANABAENA CIANOBAKTÉRIUM NEMZETSÉGBE SOROLT TÖRZSEINEK MOLEKULÁRIS TAXONÓMIAI JELLEMZÉSE Készítette: Horváth Nándor Mosonmagyaróvár 2020 A MOSONMAGYARÓVÁRI ALGAGYŰJTEMÉNY (MACC) KORÁBBAN ALAKTANI SZEMPONTBÓL ANABAENA CIANOBAKTÉRIUM NEMZETSÉGBE SOROLT TÖRZSEINEK MOLEKULÁRIS TAXONÓMIAI JELLEMZÉSE Írta: Horváth Nándor Értekezés doktori (PhD) fokozat elnyerésére Készült a Széchenyi István Egyetem Mezőgazdaság- és Élelmiszertudományi Kar, Wittmann Antal Növény-, Állat- és Élelmiszertudományi Multidiszciplináris Doktori Iskola, Haberlandt Gottlieb Növénytudományi Doktori program keretében Témavezetők: Prof. Dr. Ördög Vince egyetemi tanár, az MTA doktora Dr. habil. Molnár Zoltán, PhD, egyetemi docens Elfogadásra javaslom (igen / nem) (aláírás) A jelölt a doktori szigorlaton…………...% -ot ért el, Mosonmagyaróvár,…..…………………. a Szigorlati Bizottság elnöke Az értekezést bírálóként elfogadásra javaslom (igen /nem) Bíráló neve: Dr. …........................ …................. igen /nem (aláírás) Bíráló neve: Dr. …........................ …................. igen /nem (aláírás) A jelölt az értekezés nyilvános vitáján…..........% - ot ért el Mosonmagyaróvár,………..……………….. a Bírálóbizottság elnöke A doktori (PhD) oklevél minősítése…............................ Az EDT elnöke Tartalomjegyzék RÖVIDÍTÉSEK JEGYZÉKE ................................................................ 7 KIVONAT ................................................................................................ 9 ABSTRACT ........................................................................................... 12 1. BEVEZETÉS ..................................................................................... 15 2. CÉLKITŰZÉS ................................................................................... 17 3. IRODALMI ÁTTEKINTÉS............................................................. 19 3.1. Az Anabaena nemzetség jelentősége a gyakorlati felhasználás szempontjából ...................................................................................... 20 3.2. A cianobaktériumok taxonómiai áttekintése ................................ 23 3.3. Az Anabaena taxonok morfológiai alapú rendszerezése ............. 24 3.4. Heterociták és akinéták ................................................................ 26 3.4.1. Az akinéták szerkezete, összetétele és anyagcseréje .............. 31 3.4.2. Az akinétaképzést befolyásoló tényezők ................................ 32 3.4.3. Az akinéták szerepe a környezetünkben ................................ 33 3.5. Különböző filogenetikai markerek cianobaktériumok elkülönítésére ...................................................................................... 34 3.6. Anabaena taxonok molekuláris biológiai vizsgálatának eredményei .......................................................................................... 36 3.7. A polifázikus megközelítés .......................................................... 40 3.8. Általános kihívások az Anabaena azonosításában ....................... 41 3.9. Anabaena-hoz hasonló nemzetségek morfológiai és filogenetikai jellemzői .............................................................................................. 43 3.10. Mikroalgák bioaktív vegyületei ................................................. 54 4. ANYAG ÉS MÓDSZER ................................................................... 57 4.1. A törzsek szaporítása .................................................................... 57 4.2. Alaktan ......................................................................................... 60 4.3. DNS kivonás, PCR, szekvencia analízis ...................................... 61 4.4. Akinéta képzés előidézése ............................................................ 64 5. EREDMÉNYEK ................................................................................ 66 4 5.1. Nostoc csoport .............................................................................. 69 5.2. Trichormus csoport: ..................................................................... 72 5.2.1. Talaj alcsoport ....................................................................... 73 5.2.2. Tavi alcsoport ........................................................................ 76 5.3. Desmonostoc csoport ................................................................... 78 5.4. Bioaktív törzsek ............................................................................ 81 5.5. A „megoldatlan” törzsek .............................................................. 82 5.6. Hatvanhárom MACC törzs alaktani vizsgálata ............................ 87 5.6.1. Trichormus csoport – Tavi alcsoport .................................... 87 5.6.2. Trichormus csoport – Talaj alcsoport .................................... 97 5.6.3. Nostoc csoport ..................................................................... 105 5.6.4. Desmonostoc csoport ........................................................... 110 5.7. Az akinétaképzés eredménye ..................................................... 114 6. AZ EREDMÉNYEK MEGVITATÁSA ........................................ 117 6.1. Filogenetikai megfigyelések ...................................................... 117 6.2. Alaktani megfigyelések .............................................................. 119 6.3. A cianobaktériumok rendszerezésénél alkalmazott nevezéktani szabályok ........................................................................................... 122 6.4. Más gyűjtemények tapasztalatai ................................................. 125 7. KÖVETKEZTETÉSEK ................................................................. 128 8. ÖSSZEFOGLALÁS ........................................................................ 130 9. IRODALOMJEGYZÉK ................................................................. 132 9.1. Tudományos közlemények hivatkozásai .................................... 132 9.2. Internetes hivatkozások .............................................................. 163 10. ÚJ TUDOMÁNYOS EREDMÉNYEK........................................ 164 KÖSZÖNETNYILVÁNÍTÁS ............................................................ 166 AZ ÉRTEKEZÉS TÉMAKÖRÉBEN MEGJELENT TUDOMÁNYOS KÖZLEMÉNYEK ................................................ 167 MELLÉKLETEK................................................................................ 170 A laboratóriumi tenyésztéséhez felhasznált Z8 tápoldat összetétele 170 5 Hasonlósági matrix ............................................................................ 171 Szekvencia különbözőség becslés ..................................................... 173 Alaktani vizsgálatok .......................................................................... 175 A filogenetikai fa összecsukott részei tételesen felsorolva ............... 180 6 RÖVIDÍTÉSEK JEGYZÉKE MACC – Mosonmagyaróvár Algal Culture Collection – Mosonmagyaróvári Algagyűjtemény, Magyarország NCBI ITS – Internal transcribed spacer - belső átíródó nemkódoló szekvencia rbclx – RubisCO enzimet – (Ribulóz-1,5-biszfoszfát-karboxiláz-oxidáz), egy széndioxid és oxigén megkötésére is képes enzimet – kódoló gén rpoC1 – RNS polimeráz C1 kloroplasztisz gén 16S rRNS – prokarióták riboszómális RNS kis alegysége 18S rRNS – eukarióták riboszómális RNS kis alegysége ACOI – Algoteca de Coimbra / Coimbra Collection of Algae – Coimbrai Algagyűjtemény, Portugália AVOC – Algal Volatile Organic Compounds – algák illékony szerves vegyületei BLAST – Basic Local Alignment Search Tool – on-line szekvenciakereső alkalmazás CCALA – Culture Collection of Autotrophic Organisms – Autotróf Szervezetek Gyűjteménye, Csehország CCAP – Culture Collection of Algae and Protozoa – Alga és Protozoa Gyűjtemény, Egyesült Királyság DNS – dezoxi-ribonukleinsav IPPAS – Culture Collection of Microalgae IPPAS – IPPAS Mikroalga Gyűjtemény, Oroszország MEGA – Molecular Evolutionary Genetics Analysis – szekvenciaelemző program 7 MUSCLE – MUltiple Sequence Comparison by Log-Expectation – szekvenciaösszehasonlító program NIES – Microbial Culture Collection at the National Institute for Environmental Studies – Japán Környezettudományi Intézet Mikroalgagyűjteménye OTU – Operational Taxonomic Unit - működési taxonómiai egység PCC – The Pasteur Culture collection of Cyanobacteria – A Pasteur- intézet Cianobaktérium Gyűjteménye PCR – Polymerase Chain Reaction – polimeráz láncreakció PGR – Plant Growth Regulator – növényi növekedésszabályozó anyag RAxML – Randomized Axelerated Maximum Likelihood – filogenetikai elemzést végző program RNS – ribonukleinsav SAG – Sammlung von Algenkulturen at University of Goettingen – Göttingeni Algagyűjtemény, Németország TIM2 + G + I model – Transition model – evolúció elemzésére használt model UTEX – Culture Collection of Algae at The University of Texas – Texasi Egyetem Algagyűjteménye Zehnder-8 – Zehnder-8 tápoldat (Zehnder
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