tailieunhanh - Clonal propagation and cryogenic storage of virus-free grapevine (Vitis vinifera L.) via meristem culture
A protocol for production of virus-free Vitis vinifera using meristem culture was developed. Meristems ( mm) of V. vinifera infected with grapevine fanleaf virus and grape leafroll associated viruses were excised from 1-yearold growing vines. | Research Article Turk J Agric For 35 (2011) 173-184 © TÜBİTAK doi: Clonal propagation and cryogenic storage of virus-free grapevine (Vitis vinifera L.) via meristem culture Mohamad SHATNAWI1,*, Ghandi ANFOKA1, Rida SHIBLI2, Mohammed AL-MAZRA'AWI1, Wesam SHAHROUR1, Areej AREBIAT1 1 Faculty of Agriculture Technology, Al-Balqa Applied University, Al-Salt 19117 - JORDAN 2Faculty of Agriculture, Jordan University of Science and Technology, Irbid - JORDAN Received: Abstract: A protocol for production of virus-free Vitis vinifera using meristem culture was developed. Meristems ( mm) of V. vinifera infected with grapevine fanleaf virus and grape leafroll associated viruses were excised from 1-yearold growing vines. Shoot tips were cultured on half-strength Murashige and Skoog (MS) medium, supplemented with mg L-1 of benzylaminopurine (BAP) and mg L-1 of naphthalene acetic acid (NAA). BAP and kinetin resulted in differences in the number of new shoots per explant, shoot height, and number of new leaves per explant. BAP at mg L-1 gave the highest in vitro multiplication rate, with shoots per explant, whereas elongation was greatest in the presence of mg L-1 of kinetin. Root initiation was tested on an MS medium supplemented with , , , , , or mg L-1 of IBA (indole-3-butyric-acid), IAA (indole-3-acetic-acid), or NAA. Maximum root number was achieved using mg L-1 of IBA. A survival rate of 95% was achieved when rooted explants were acclimatized ex vitro in a mixture of 1 soil: 1 perlite: 1 peat. Acclimatized plants grew in the greenhouse and were maintained as virus-free plants. Visual inspections as well as results of RT-PCR, using virus-specific oligonucleotide primers, showed that plants developed in vitro were free from grapevine fanleaf virus, grape leafroll associated virus-1, and grape leafroll associated virus-3 infections. Shoot tips from plantlets grown in vitro were cryopreserved
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