Date Received: 05-06-2025
Date Published: 13-06-2025
##submissions.doi##: https://doi.org/10.31817/tckhnnvn.2012.10.2.
Views
Downloads
How to Cite:
Virus Cleaning of Galic, in Allium Sativum L., by Meristem Culture
,
Nguyen Thi Ly Anh
Keywords
Allium sativum L., meristem, regeneration, micropropagation, virus-infected, RT-PCR
Abstract
A suitable protocol for meristem-tip culture for garlic (Allium sativum L.) to eliminate viruses was studied and established. The presence of viruses were identified by: (i) mechanical inoculation of inoculum isolated from infected garlic plants on indicator Chenopodium album L. and (ii) RT-PCR using potyvirus degenerate primers. After pretreatment with ethanol (70%) for 1 minute, shoots from infected plants were further treated with sodium dichloroisocyanurate (5g/l) for 5 minutes to eliminate microorganisms. Under a microscope, leaves of infected plants were removed one-by-one and meristems were excised and grown on MS+30g saccarose + 1mg BA/l to produce shoots. It is indicated that to get completely virus-free plants, meristems shoud be excised with size below 0.3 mm in length. If the exised meristem explants are of 1mm in size the culture medium should be added with ribavirin (20mg/l). The virus-free shoots that were multiplied on MS + 30g saccarose + 0,5mg αNAA/l + 2,0mg BA/l media yield not only the highest regeneration rate (4.08 times) but also the best growth (7.55cm). Adventitious shoots can also be regenerated via calluses on MS + 15g saccarose/l + 10g glucose/l + 5g manitol/l + 2,0mg BA/l media. The best medium for root generation was MS + 30g saccarose + 0,5g activated charcoal/l + 0,5mg αNAA/l, on which, 100 % shoots produced adventitous roots (5.19 roots per shoot on avarage) and the resulting plants grew normally.
References
Tạ Thu Cúc, Hồ Hữu An, Nghiêm Thị Bích Hà (2000). Giáo trình cây rau, NXB Nông nghiệp, Hà Nội, Tr 262 - 233.
A.S. Abdel Wahab, S. Elnagar and M.A.K. El-Sheikh (2009). Incidence of Aphid-borne onion yellow dwarf virus (OYDV) in Alliaceae Crops and Asociated Weeds in Egypt 4 th Conference on Recent Technologies in Agriculture, pp 21-23.
Barg, E., Lesemann, D.E and Vetten, H.J (1994). Identification, partial characterization, and distribution of viruses infecting Allium crops in south and southeast Asia. Acta Hor-tic.358:251- 258.
Bos, L. (1976). Onion yellow dwarf virus. CMI (Commonw. Mycol. Inst.)/AAB (Assoc. Appl. Biol.) Descriptions of Plant Viruses. No. 158. Kew, Surrey, England.
Bos, L., Huijberts, N., Huttinga, H., and Maat, D. Z. (1978). Leek yellow stripe virus and its relationships to onion yellow dwarf virus; characterization, ecology and possible control. Neth. J. Plant Pathol 84:185-204.
Diekmann M, (1995). Allium spp. FAO/IPGRI Technical Guidelines for the Safe Movement of Germplasm, pp 19-31.
Gisele Irvine and Samantha Sterling (2002). Increasing the export potential - opportunities for Australian garlic, pp 1-3.
Ha, C.V., Coombs, S., Revill1, P.A., Harding, R.M., Vu, M.T. and Dale, J.L. (2008a). Identification and sequence analysis of potyviruses infecting crops in Vietnam. Archives of virology. Vol.153, p:45-60.
Ha, C.V., Coombs, S., Revill1, P.A., Harding, R.M., Vu, M.T. and Dale, J.L. (2008b). Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses. Archives of virology. Vol.153, p:25-36.
Vũ Triệu Mân, Lê Lương Tề (2001). Giáo trình bệnh cây nông nghiệp, NXB Nông nghiệp, Hà Nội.
Lot H., Chovelon V., Souche S. and Delecolle,B. (1998). Effects of onion yellow dwarf and leek yellow stripe viruses on symptomatology and yield loss of three French garlic cultivars. Plant-Disease., pp 82-(12):1381-1385
Schwartz H. F. and Mohan S.K. (1995). Compendium of onion and garlic diseases. APS. PRESS. The American Phytopathological Society. pp 52.
Van der Vlugt, R.A.A., Steffens, P., Cuperus, C., Barg, E., Lesemann, D.E., Bos, L., and Vetten, H.J. (1999). Further evidence that shallot yellow stripe virus (SYSV) is a distinct potyvirus and reidentification of Welsh onion yellow stripe virus as a SYSV strain. Phytopathology. Vol.89, p: 148-155.