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A circular DNA molecule, designated as DNAβ, was identified in tobacco plants infected with Tobacco leaf curl virus (TLCV) isolates Y5 and Y8 by PCR using primers based on the conserved region of the two reported DNAβsequences of whitefly-transmitted geminiviruses (WTGs). The complete nucleotide sequences of DNAβof Y5 and Y8 (TLCV DNAβ) were determined. Y5 DNAβcomprises 1333 nucleotides encoding 8 predicted ORFs with 4 ORFs in virion-sense DNA and 4 ORFs in complementary-sense DNA; Y8 DNAβconsists of 1338 nucleotides encoding 7 predicted ORFs with 4 ORFs in virion-sense DNA and 3 ORFs in complementary-sense DNA. TLCV DNAβhas little sequence homology to DNA-A of TLCV, except that it shares conserved TAATATTAC loop sequence with TLCV DNA-A. Sequence comparison showed that Y5 DNAβshared 85% sequence homology with Y8 DNAβ, and both Y5 DNAβand Y8 DNAβhad relatively low sequence identity (51%-65%) with the reported DNAβmolecules associated with Ageratum yellow vein virus and Cotton leaf curl virus. The immunotrapping PCR and whitefly transmission tests showed that DNAβmolecule could be encapsidated in virus particle and transmitted by Bemisia tabaci. This is the first report of DNAβassociated with WTGs in China.