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Naslov:EKSPRESIJA GENOV ZA EVOLUCIJO ETILENA TER ABSCIZIJA PLODIČEV JABLANE (Malus domestica Borkh.) PO TRETIRANJU Z RASTNIMI REGULATORJI IN SENČENJEM
Avtorji:ID Kolarič, Jure (Avtor)
ID TOJNKO, Stanislav (Mentor) Več o mentorju... Novo okno
Datoteke:.pdf DR_Kolaric_Jure_2011.pdf (2,97 MB)
MD5: CC22B288D56A599EC158209BF719B877
PID: 20.500.12556/dkum/07eed57e-e693-43dc-bb3e-345bbf563591
 
Jezik:Slovenski jezik
Vrsta gradiva:Doktorska disertacija
Organizacija:FKBV - Fakulteta za kmetijstvo in biosistemske vede
Opis:Kljub temu da je naravni mehanizem junijskega odpadanja plodičev pri jablani (Malus domestica Borkh.) dobro razvit, je potrebno za doseganje redne rodnosti in boljše kakovosti plodov uporabljati tehnološke ukrepe, s katerimi dodatno vzpodbudimo odpadanje plodičev. Uveljavljena metoda za redčenje plodičev je škropljenje dreves z rastnimi regulatorji, čeprav mehanizem delovanja omenjenih sredstev še vedno ni jasen. Pomemben dejavnik pri razvoju abscizijskih procesov je tudi rastlinski hormon etilen, vendar zaenkrat njegova vloga še ni popolnoma raziskana. Podobno kot z rastnimi regulatorji (NAA, BA in ETEF) lahko na abscizijo vplivamo tudi s senčenjem. Namen raziskave je bil podrobneje proučiti vlogo etilena v procesu abscizije plodičev ter ugotoviti, ali je prisotnost etilena neodvisna od dejavnika, s katerim sprožimo abscizijo (rastni regulatorji, senčenje). Proučevali smo povezavo med ekspresijo genov, odgovornih za evolucijo etilena (MdACO1, MdACS5A in MdACS5B), in dejansko frekvenco odpadanja oz. deležem plodov, ki so bili podvrženi absciziji. Glede na dejstvo, da so lateralni plodiči (LP) bolj podvrženi abscizijskim procesom v primerjavi s centralnimi plodiči (CP) smo želeli ugotoviti, ali obstajajo razlike tudi pri izraženosti opazovanih genov. Za izvedbo raziskave smo v jablanovem nasadu na sorti 'Zlati delišes' zasnovali dvoletni poljski poskus v poskusnem nasadu Kmetijskega inštituta Slovenije na Brdu pri Lukovici. Poskus je bil razdeljen na dva dela, od katerih je vsak zajemal 8 naključnih blokov s 5 obravnavanji. Tretiranje z rastnimi regulatorji in senčenje smo opravili, ko je bil povprečen premer plodičev ≈10 mm. V obeh letih smo imeli sledeča obravnavanja: 1. K (netretirano), 2. NAA (15 mg l-1), 3. BA (150 mg l-1), 4. ETEF (500 mg l-1) in 5. SEN (3 dni, 96 % redukcija fotosintetsko aktivnega sevanja). V času trajanja poskusa smo na drevesih oz. plodičih opravljali različne meritve, in sicer: prirast plodičev, frekvenco odpadanja plodičev, evolucijo etilena, vzorčenje tkiva za genetske analize in končno vrednotenje pridelka po posameznem drevesu. Evolucijo etilena smo spremljali v terminih 0, 2, 5 in 8 dni po tretiranju (DAT), in sicer smo koncentracijo etilena v vzorcih določili s plinsko kromatografijo (GC). V letu 2008 smo vzorce za genetske analize vzorčili 0 in 8 DAT, medtem ko v letu 2009 v terminih 0, 1, 4 in 7 DAT. Za genetske analize, ki smo jih opravili v laboratorijih Kmetijskega inštituta Slovenije, smo vzorčili tkivo CP in LP ter tkivo abscizijske cone obeh tipov plodičev. Najprej smo iz vzorcev izolirali celokupno RNA, nato pa smo za opazovanje izražanja genov MdACS5A, MdACS5B in MdACO1 uporabili kvantitativno verižno reakcijo s polimerazo (qPCR). V času tehnološke zrelosti smo še ovrednotili pridelek po posameznem drevesu. Nanos NAA, BA in ETEF je v letih 2008 in 2009 povzročil večjo abscizijo plodičev v primerjavi s K. Podoben učinek smo v letu 2009 dosegli s SEN, medtem ko v letu 2008 SEN ni vplivalo na abscizijo plodičev. Potrdili smo, da so bili absciziji mnogo bolj podvrženi LP pri vseh obravnavanjih, medtem ko je nekoliko večjo abscizijo CP povzročila samo aplikacija ETEF. Spremljanje priraščanja plodičev po aplikaciji rastnih regulatorjev in senčenju je pokazalo, da ETEF najmočneje zavira rast plodičev, tudi v primeru ko ti niso podvrženi kasnejši absciziji. Medtem pa NAA, BA, in SEN zavrejo predvsem rast plodičev, ki so pozneje podvrženi absciziji. Neglede na obravnavanje pa smo ugotovili, da je rast plodičev pred abscizijo v vseh primerih zavirana oz. zaustavljena. To pomeni, da je tudi rast plodičev oz. njeno zaviranje pokazatelj abscizije. Evolucija etilena iz plodičev je bila različna med posameznimi obravnavanji, prav tako pa je imel vpliv tudi tip plodičev. Tako smo v primeru CP najmočnejšo evolucijo etilena izmerili pri obravnavanju ETEF, ki se je povišala v terminu 5 in 8 DAT, medtem pa ostala obravnavanja niso povzročila večje evolucije etilena v CP. Nasprotno pa smo pri LP, poleg obra
Ključne besede:abscizija, evolucija etilena, redčenje, rastni regulatorji, genetska ekspresija, qPCR
Kraj izida:Maribor
Leto izida:2011
PID:20.500.12556/DKUM-20899 Novo okno
NUK URN:URN:SI:UM:DK:VJ6J8FS7
Datum objave v DKUM:17.10.2011
Število ogledov:4952
Število prenosov:389
Metapodatki:XML DC-XML DC-RDF
Področja:FKBV
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Sekundarni jezik

Jezik:Angleški jezik
Naslov:EXPRESSION OF GENES ASSOCIATED WITH ETHYLENE EVOLUTION AND ABSCISSION OF APPLE (Malus domestica Borkh.) FRUITLETS AFTER PLANT GROWTH REGULATORS AND SHADING TREATMENTS
Opis:In spite of the fact that the natural mechanism of June drop is well developed in apple (Malus domestica Borkh.), in order to obtain regular bearing and higher amount of the most valuable fruit, thinning as a technological measure cannot be avoided. The use of plant growth regulators is an established measure for fruit set regulation, although the mode of action of regulators is unclear. An important factor in the development of abscission process is the gaseous hormone ethylene, but until now its role has not been fully explored yet. Similarly as with growth regulators, abscission can be induced by shading. The aim of our research was to analyze the role of ethylene in the fruitlet abscission process and to find out if ethylene was involved in abscission regardless of factors which induced the process (growth regulators, shading). We also studied the relation between the expression of genes related to ethylene biosynthesis (MdACO1, MdACS5A and MdACS5B) and actual abscission dynamics, and the amount of abscised fruitlets. Taking into account that lateral fruitlets (LF) are more subjected to abscission in comparison with the king fruitlets (KF) we wanted to find out if there were differences between LF and KF also in the case of expression of observed genes. In the experimental orchard of Agricultural Institute of Slovenia situated at Brdo near Lukovica, a two-year field trial was designed involving the apple variety ‘Golden Delicious’. The experiment was divided into two groups, and each group consisted of a complete randomized block experiment with eight replications and five treatments (K, NAA, BA, ETEF and SEN). During the experiment, different measurements were performed on trees and fruitlets, namely: fruit growth, abscission dynamics, ethylene evolution, sampling of tissue for molecular analysis and evaluation of yield per tree at harvest. The ethylene evolution was observed 0, 2, 5 and 8 days after treatment (DAT) and it was based on gas chromatography (GC). The samples for molecular analysis were collected in 2008 at 0 and 8 DAT, but in 2009, sampling dates were 0, 1, 4, and 7 DAT. The molecular analyses were performed in the laboratory of the Agricultural Institute of Slovenia. In the first step, the total RNA was isolated from each sample and then, after conducting quantitative polymerase chain reaction (qPCR), we assessed the expression of MdACS5A, MdACS5B and MdACO1. The application of NAA, BA and ETEF, in 2008 and 2009, effectively increased the number of abscised young fruits. Similar effect was achieved in 2009 by shading, but not in 2008. We believe that the most probable reason for ineffectiveness of shading in 2008 were relatively low temperatures when measurements took place. We also confirmed that LF were more subjected to abscission while in the case of KF only the ETEF treatment induced it. The strongest nonselective inhibiting effect on fruit growth was recorded after the ETEF application while NAA, BA and SEN inhibited mostly fruitlets which were later subjected to abscission process. Results of ethylene evolution showed that, next to treatment, the type of fruitlets (i.e. KF or LF) was an important factor. In the case of KF only ETEF enhanced the ethylene evolution, since in the case of LF also NAA, BA, and SEN influenced the ethylene biosynthesis. The greatest differences between sampling dates and treatments were recorded in the expression of MdACO1 while changes in the expression of MdACS5A and MdACS5B were at a much lower level. The greatest influence on the expression of the observed genes was caused by the ETEF treatment, which was the only treatment that increased MdACO1 in the tissues of KF. Similarly, only the ETEF treatment increased the expression of all observed genes in the abscission zone tissue. In the case of LF, all treatments, except the K, enhanced the expression of MdACO1. The expression of MdACO1 in fruitlet tissue was found to be a reliable indicator of abscission, although the amount of the target gene was not related with the amount of the abscised frui
Ključne besede:abscission, ethylene evolution, thinning, growth regulators, gene expression, qPCR


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