Chinese tulip tree (Liriodendron chinensis) is native to China and is planted all around the country as an ornamental tree. In July of 2011, some Chinese tulip trees with typical phytoplasma symptoms were found in Baoding City, Hebei Province, China. Symptoms included yellowing of leaves, slow decline, little leaves, and death of entire plants. To confirm phytoplasma infection of these plants, total DNA was extracted from 100 mg of fresh leaf midribs collected from leaves of nine symptomatic and eight asymptomatic plants with a plant DNA extract kit (Tiangen, Beijing, China) according to the manufacturer's protocols. Using 16S rRNA phytoplasma universal primer pairs P1/P7 followed by R16F2n/R16R2, a nest PCR was carried out (1,2). The results showed that the phytoplasma was only detected in symptomatic samples by nested PCR, while the asymptomatic were negative. An approximate 1.2-kb specific fragment was obtained from the DNA of nine symptomatic plants, but no product was amplified from the leaves of eight healthy ones. The amplified products were cloned and sequenced. The sequence was deposited in GenBank Data Libraries under Accession No. JQ585925 and shared the highest homology of 99% with Puna chicory flat stem phytoplasma (GenBank Accession No. JN582266), Apricot leaf roll phytoplasma (GenBank Accession No. FJ572660), Jujube witches'-broom phytoplasma (GenBank Accession No. AY197661), and other elm yellows group phytoplasmas by BLAST analysis with that of other phytoplasmas from GenBank. Meanwhile, the sequence data was analyzed by iPhyClassifier software and the result showed that the 16S rDNA F2nR2 fragment was identical (similarity coefficient 1.00) to the reference patterns of 16Sr group V, subgroup B (GenBank Accession No. AB052876) (3). Combining the BLAST analysis in GenBank and the analysis of iPhyClassifier, we classified the phytoplasma causing Chinese tulip tree yellow leaves disease into subgroup 16SrV-B. To our knowledge, this is the first report of the 16SrV-B group phytoplasmas infecting Chinese tulip tree in China.
References: (1) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) I. M. Lee et al. Int. J. Syst. Evol. Microbiol. 54:337, 2004. (3) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.
