Phytoplasma associated with Chinese tallow tree yellowing disease in China represents a new 16SrIII subgroup

Abstract

Summary Recently, samples of the Chinese tallow tree (Sapium sebiferum) displaying yellowing symptoms were collected from a grove in Tai'an, Shandong Province, China. The association of phytoplasma with yellowing disease was ascertained using nested polymerase chain reaction (PCR) of the 16S rRNA gene by using the phytoplasma‐specific universal primer pair P1/P7, followed by R16F2n/R16R2 as nested primers, and rp genes primed using rpL2F3/rp(I)R1A followed by rp(III)‐FN/rp(I)R1A. The sequence and phylogenetic analyses of the 16S rRNA gene and rp genes revealed that the phytoplasma associated with the Chinese tallow tree belonged to the 16SrIII group (the X‐disease group). Computer‐simulated and gel‐based restriction fragment length polymorphism (RFLP) analyses revealed that the RFLP patterns were different from the reference patterns of all previously established 16SrIII subgroups, with the maximum similarity coefficient exceeding the threshold for delineation of a new subgroup RFLP pattern type within the 16SrIII group. Thus, the phytoplasma associated with the Chinese tallow tree yellowing disease, designated as ‘CTTY’, represents a new subgroup (16SrIII‐Y). This study shows the Chinese tallow tree as a new host of phytoplasma belonging to the 16SrIII group in China and worldwide.

Identification of ‘Candidatus phytoplasma ziziphi’ associated with persimmon (Diospyros kaki Thunb.) fasciation in China

“…Nested PCR assays for phytoplasma 16S rRNA gene were performed with the universal primers P1/P7 (Schneider et al 1995), followed by the R16F2n/R16R2 primers (Gundersen-Rindal and Lee 1996) after 1 : 20 dilution of the products of the direct PCR. The parameters of the PCR mixtures and amplification programs for nested PCRs were described previously (Gao et al 2014). The purified R16F2n/R16R2 PCR amplification products were cloned into the pMD18-T vector (Takara Bio Inc., Dalian, China) following the manufacturer's instructions and then transferred into Escherichia coli DH5a competent cells.…”

“…1). The parameters of the PCR mixtures and amplification programs for nested PCRs were described previously (Gao et al 2014). Nested PCR assays for phytoplasma 16S rRNA gene were performed with the universal primers P1/P7 (Schneider et al 1995), followed by the R16F2n/R16R2 primers (Gundersen-Rindal and Lee 1996) after 1 : 20 dilution of the products of the direct PCR.…”