INOCULATION OF SUGAR-BEET PROTOPLASTS WITH BEET NECROTIC YELLOW VEIN VIRUS-PARTICLES BY MILD SONICATION

被引:10
作者
JOERSBO, M
BRUNSTEDT, J
机构
[1] DANISCO A/S, Biotechnology Research Division, Copenhagen
关键词
Beet necrotic yellow vein virus; Protoplast; Sugar beet; Ultrasound; Virus infection;
D O I
10.1016/0166-0934(90)90008-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A novel procedure employing mild sonication for in vitro inoculation of plant protoplasts with virus particles has been established. Sugar beet (Beta vulgaris L.) protoplasts were briefly exposed to 20 kHz ultrasound in the presence of beet necrotic yellow vein virus (BNYVV) particles. The accumulation of BNYVV coat protein was analyzed by an enzyme-linked immunosorbent assay. The infection was detectable after 16 h and reached maximum 3-3 1 2 days after inoculation. Maximum levels of BNYVV coat protein in inoculated protoplasts were obtained by sonication for 500-600 ms at 45-60 W. This reduced the viability to 15-30%. The efficiency of the inoculation increased with the concentration of BNYVV particles up to 28 μg/ml. Infection was found to be optimal when the BNYVV particles were added just before sonication but low levels of infection could be obtained by addition of virus particles up to 60 min after sonication. © 1990.
引用
收藏
页码:63 / 69
页数:7
相关论文
共 16 条
  • [1] CHAPMAN IV, 1974, BRIT J RADIOL, V47, P411, DOI 10.1259/0007-1285-47-559-411
  • [2] FECHHEIMER M, 1987, METHOD CELL BIOL, V28, P179
  • [3] ISOLATION, CULTURE AND REGENERATION OF PETUNIA LEAF PROTOPLASTS
    FREARSON, EM
    POWER, JB
    COCKING, EC
    [J]. DEVELOPMENTAL BIOLOGY, 1973, 33 (01) : 130 - 137
  • [4] Fujisawa I., 1977, Annals of the Phytopathological Society of Japan, V43, P583, DOI 10.3186/jjphytopath.43.583
  • [5] LIPOSOME-MEDIATED INFECTION OF PLANT-PROTOPLASTS WITH TOBACCO MOSAIC-VIRUS RNA
    FUKUNAGA, Y
    NAGATA, T
    TAKEBE, I
    [J]. VIROLOGY, 1981, 113 (02) : 752 - 760
  • [6] JOERSBO M, 1990, IN PRESS PLANT CELL
  • [7] POLYETHYLENE GLYCOL-MEDIATED INFECTION OF CUCUMBER PROTOPLASTS BY CUCUMBER MOSAIC-VIRUS AND VIRUS-RNA
    MAULE, AJ
    BOULTON, MI
    EDMUNDS, C
    WOOD, KR
    [J]. JOURNAL OF GENERAL VIROLOGY, 1980, 47 (MAR) : 199 - 203
  • [8] INFECTION OF TOBACCO PROTOPLASTS WITH COWPEA CHLOROTIC MOTTLE VIRUS AND ITS RNA
    MOTOYOSHI, F
    BANCROFT, JB
    WATTS, JW
    BURGESS, J
    [J]. JOURNAL OF GENERAL VIROLOGY, 1973, 20 (AUG) : 177 - 193
  • [9] MOTOYOSHI F, 1974, J GEN VIROL, V2, P89
  • [10] ELECTROPORATION-MEDIATED INFECTION OF TOBACCO LEAF PROTOPLASTS WITH TOBACCO MOSAIC-VIRUS RNA AND CUCUMBER MOSAIC-VIRUS RNA
    NISHIGUCHI, M
    LANGRIDGE, WHR
    SZALAY, AA
    ZAITLIN, M
    [J]. PLANT CELL REPORTS, 1986, 5 (01) : 57 - 60