An Uncultured Bacterium Associated With Infection in Brassica Oleracea Italica in Malatya Province


Usta M., Oksal H. D., Kaya Apak F., Altuntaş Ö., Poyraz P., Basım H., ...More

The International Conference on Agriculture, Forest, Food Sciences and Technologies(ICAFOF), Nevşehir, Turkey, 15 - 17 May 2017, pp.1167

  • Publication Type: Conference Paper / Full Text
  • City: Nevşehir
  • Country: Turkey
  • Page Numbers: pp.1167
  • Van Yüzüncü Yıl University Affiliated: Yes

Abstract

Broccoli (Brassica oleracea Italica) plants with symptoms of leaf curling, enation, fasciation, dwarfing, vain banding, swelling and cracking symptoms were observed in greenhouse grown plants in Malatya province in 2016.  Genomic  DNA isolation was made  from leaf tissue and midrib  of  symptomatic  and  non-symptomatic  plants.  By  using  R16mF2/R16mR1  and R16F2n/R16R2  primers 16S  rDNA  fragments  were  subjected  to  nested  polymerase  chain reaction  (Nested-PCR)  for  the  involvement  of  possible  phytoplasma  infection.  Symptomatic plants  were  yielded  approx.  1.2-kb  DNA  fragments.  Amplified  PCR  fragments  were  cloned into a proper plasmid vector and transformed into competent Escherichia coli strain JM 109. Recombinant  plasmid  DNA  was  isolated  and  sequenced  bi-directionally.  The  presence  of white  fly  was  observed  on  and  around  the  symptomatic  plants.  Fifteen  white  flies  were sampled  from  greenhouse  to  assess  the  presence  of  the  suspected  agent.  The  genomic  DNA was  isolated  and  subjected  to  Nested-PCR  by  using  same  primer  pairs.  None  of  the  tested white  fly  was  reacted  positive  for  the  presence  of  expected  DNA  fragments. Based  on sequence  data,  phylogeny  analysis  and virtual  restriction  fragment  length  polymorphism (RFLP)  analysis,  an  unidentified  and  uncultured  bacterium  has  been  reported  in  broccoli plants in Malatya. The attempts to visualize the agent by electron microscopy in isolated host tissues are still in progress.