Cloning and Nucleotide Sequence Analysis of IS Gene of Potato Leaf Roll Virus Inner Mongolia Isolate

  • GUO Zhi-hua ,
  • SUN Yi ,
  • ZHANG Xiao-mei ,
  • ZHANG Jian ,
  • BAI Yun-feng
Expand
  • 1. Department of Bioengineering, Shanxi University, Taiyuan 030006, China;
    2. Shanxi Academy of Agricultural Science, Taiyuan 030031, China

Received date: 2007-09-11

  Online published: 2014-10-14

Abstract

With a pair of specfric primers based on Potato Leaf Roll Virus(Pb)genomic conservative sequence lying on the both side of Intergenic Sequence(IS) reported in Genbank, one cDNA fragment(369 bp) containing the IS was amplified by reverse transcription polymerase chain reaction(RT-PCR) using the total RNA of PbV Inner Mongolia isolate as a template. The fragment was cloned into pBS-T vector and identified by PCR, restrictive enzyme analysis and nucleotide sequence analysis. The cloned IS sequence was compared with that of homologous gene of all other 13 available PbV isolates. The results showed that it had high homology with the other isolates(the highest homology reached 100% of nucleic acid and the average homology was 97. 90%, while the homology of 13 PbV complete genome was 96. 81%).The IS was not only conservative anong PbV strains but also conservative in the sequences of PbV complete genome. The result implied that transgen cplant with high resistance to PbV could be obtained through transfening RNA interfere structure of IS into potatoes.

Cite this article

GUO Zhi-hua , SUN Yi , ZHANG Xiao-mei , ZHANG Jian , BAI Yun-feng . Cloning and Nucleotide Sequence Analysis of IS Gene of Potato Leaf Roll Virus Inner Mongolia Isolate[J]. Acta Agriculturae Boreali-Sinica, 2007 , 22(6) : 19 -23 . DOI: 10.7668/hbnxb.2007.06.005

References

[1] KOJ IMA M. Purification and electron microscopy of Potato Leafroll Virus[J]. Virology, 1969, 39: 162-171.
[2] 张鹤龄. 马铃薯卷叶病毒(PLRV)基因组研究进展[J]. 中国病毒学, 1996, 11(1): 1-8.
[3] Robert R, Martin Paul K, Keese Mark J, et al. Evolution and molecular biology ofLuteoviruses[J]. Phytopathology, 1990, 28: 341-363.
[4] Murphy F A, Fauquet C M, Bishop D H L, et al. Virus taxonomy classification and nomenclature of viruses, Report of the International Committee[J]. Archives of Virology, 1995, 10: 187-192.
[5] Mayo M A, Robinson K J, Barker H. In Report of the Scottish Crop Research Institute for 1983[R]. 1984: 187.
[6] Mayo M A, Robinson D J, Jolly C A, et al. Nucleotide of potato leafrolll Luteovirus RNA[J]. Gen Virol, 1990, 70: 1037-1051.
[7] Tenllado F, Diaz-Ruiz J R. Double-strand RNA-mediated interference with plant virus infection[J]. J Virology, 2001, 75(24): 12288-12297.
[8] Mueller E, Gilbert J, Davenport G, et al. Homology-dependent resistance, transgenic virus resistance in plants related to homology-dependent gene silencing[J]. Plant, 1995, 7: 1001-1013.
[9] Sambrook J, Russell D W. Molecular Cloning: A Laboratory Manual(3rd ed)[M]. New York: Cold Spring Harbor Laboratory Press, 2001.
[10] Sanford J C, Johnston S A. The concept of parasite-derived resistance-deriving resistance genes from the parasite′s own genome[J]. Theor Biol, 1985, 113: 395-405.
[11] 白云凤, 赵晋锋, 郑军, 等. 反义外壳蛋白基因介导的抗SCMV转基因玉米研究[J]. 作物学报, 2006, 33(5): 661-665.
[12] Smith N A, Singh S P, Wang M B, et al. Total silencing by intron-spliced hairpin RNAs[J]. Nature, 2000, 407: 319-320.
[13] Wang M B, Abbott D C, Waterhouse P M. A single copy of a virus-derived transgene encoding hairpin RNA gives immunity to barley yellow dwarf virus[J]. Mol Plant Pathol, 2000, 1(60): 347-356.
[14] Missiou A, Kalantidis K, Boutlal A, et al. Generation of transgenic potato plants highly resistant to potato virus Y(PVY)through RNA silencing[J]. Molecular Breeding, 2004, 14: 185-197.
[15] 白云凤, 杨红春, 曲琳, 等. 抗甘蔗花叶病毒的无标记反向重复转基因玉米[J]. 作物学报, 2007, 33(6): 973-978.
Outlines

/