Molecular Characterization And Functional Analysis Of A New Acid Phosphatase Gene (Ha-acp1) From The Cereal Cyst Nematode Heterodera Avenae

The cereal cyst nematode（Heterodera avenae）, is a serious pest of cereal crops world wide. Thedistribution areas of H. avenae now includes16provinces in China. And the H.avenae now is the majorrestriction to the wheat yield. In this study a new acid phosphatase gene (Ha-acp1) expressed in thesubventral glands of the cereal cyst nematode (Heterodera avenae) was cloned and the characteristics ofthis gene were analyzed. The full length cDNA sequence of1608bp with highest homology towardsmsp21(AY134440), an acid phosphatase of Meloidogyne incoginta. Correspondingly, the gene wasnamed Ha-acp1(accession number JQ341056). The sequence contained a1455bp open reading frame(ORF) encoding484amino acids. The Ha-acp1gDNA was2627bp long from ATG to the stop codon.The intron regions were identified by aligning the genomic sequence to the corresponding cDNAsequence. Fourteen introns were identified in Ha-acp1gDNA. For sedentary endo parasitic nematodes,parasitism genes encoding secretory protein expressed in the subventral glands cells always play animportant role during the early parasitic process. Results showed that the gene has a putative signalpeptide for secretion and in situ hybridization showed that the transcripts of Ha-acp1accumulatedspecifically in the subventral gland cells of H. avenae. Southern blot analysis suggested that Ha-acp1belongs to a multigene family.RNAi technology was used to research the function of the Ha-acp1gene. We built and optimizedthe isolate system. RT PCR analysis indicated that this transcription was strong in the pre parasiticjuvenile. Knocking down of Ha-acp1using RNA interference technology can reduce nematodeinfectivity by50%, and can suppress the development of cyst. Results indicated that Ha-acp1can playan important role in destroying the defense system of the host plant.