Molecular characterization of a new member of the 16SrV group of phytoplasma associated with Bischofia polycarpa (Levl.) Airy Shaw witches’ -broom disease in China by a multiple gene-based analysis

Abstract

Bischofia polycarpa (Levl.) Airy Shaw witches'-broom disease is widespread in southern China. Infected trees show symptoms of severely proliferating shoots, reduced leaf size, yellowing and reddening of leaves, and internode shortening. We detected the phytoplasma in diseased tissues by 4′,6-diamidino-2-phenylindole-HCl fluorescence microscopy and polymerase chain reaction using the universal primers P1/ P7. We conducted sequence similarity analyses based on 16S rRNA and 23S rRNA genes, the spacer region between the 16S and 23S rRNA genes (ISR), the partial ribosomal proteinencoding operon (rp), and the elongation factor Tu (tuf) gene of the BiWB phytoplasma. These analyses unanimously showed that the BiWB phytoplasma belongs to the elm yellows group (the 16SrV group). Phylogenetic and computersimulated restriction fragment length polymorphism analyses using the nearly full-length 16S rRNA gene sequence demonstrated that the BiWB phytoplasma is more closely related to the subgroup 16SrV-B than to the other subgroups in 16SrV. It may represent a novel subgroup, designed as 16SrV-H, which is distinct from other subgroups of phytoplasmas in the 16SrV group.

Accurate and sensitive detection of areca palm yellow leaf phytoplasma in China by droplet digital PCR targeting tuf gene sequence

“…The results showed that the established ddPCR method was reliable for quantitative detection of AYL phytoplasma, with relatively good effect. The tuf gene is a functional protein coding gene and a relatively conserved sequence in the phytoplasma which plays an important role in protein translation and evolution of phytoplasma (Koui et al, 2003; Lai et al, 2014; Lee et al, 2000; Schneider et al, 1997; Yu et al, 2016,b, 2017, 2018). In the study, the tuf gene was used as a diagnostic assay for AYL phytoplasma for the first time.…”

“…The results showed that the established ddPCR method was reliable for quantitative detection of AYL phytoplasma, with relatively good effect. The tuf gene is a functional protein coding gene and a relatively conserved sequence in the phytoplasma which plays an important role in protein translation and evolution of phytoplasma (Koui et al, 2003;Lai et al, 2014;Lee et al, 2000; F I G U R E 3 Absolute quantitation of the ddPCR detection system for AYL phytoplasma using primers Atf/Atr and probe AtProbe. AYL1, AYL2 and AYL3 represent AYL samples; H1, H2 and H3 represent asymptomatic samples; ck1 represented TE buffer used for DNA extraction; ck2 represented ddH2O used for ddPCR reaction system.…”

Occurrence, Impact, and Multilocus Sequence Analysis of Alder Yellows Phytoplasma Infecting Common Alder and Italian Alder in Southern Italy

“…ALY is associated with the ALY phytoplasma, a member of the elm yellows (EY) phytoplasma group or 16SrV group, subgroup 16SrV-C [ 18 ]. Other members of this group are phytoplasmas causing diseases of trees and shrubs such as EY, flavescence dorée (FD) of grapevine, Palatinate grapevine yellows (PGY), spartium witches’-broom (SpaWB), rubus stunt (RuS), blackberry witches’-broom, cherry lethal yellows, flowering cherry decline, sweet cherry virescence, peach yellows in India, jujube witches’-broom, balanites witches’-broom, Japanese raisin witches’-broom, Bischofia polycarpa witches’-broom and Styphnolobium japonicum (previously known as Sophora japonica ) witches’-broom, and phytoplasmas infecting Clematis vitalba in Europe and hemp dogbane ( Apocynum cannabinum ) in New York state, respectively [ 2 , 18 , 19 , 20 , 21 , 22 , 23 , 24 ]. Similarly to other members of the EY group, ALY phytoplasma is a homogeneous pathogen at rDNA sequence level.…”

Multigene characterization of 'Candidatus Phytoplasma ulmi'‐related isolates associated with elm yellows disease of Ulmus minor Mill. in Poland

“…The elm yellows phytoplasma group, 16SrV, is divided into eight subgroups, 16SrV‐A to 16SrV‐I (Fránová et al, 2016) of phytoplasmas associated with serious diseases such as grapevine flavescence dorée (Martini et al, 1999), alder yellows (Lederer & Seemüller, 2007), elm yellows (Lee et al, 2004), rubus stunt (Malembic‐Maher et al, 2011), spartium witches’ broom (Marcone et al, 2004), cherry lethal yellows (Zhu et al, 1998), jujube witches’ broom (Jung et al, 2003), Balanites witches’ broom (Win et al, 2013), (Lai et al, 2014) and 16SrV‐I blackberry witches’ broom (Fránová et al, 2016). The reference strain of the elm yellows group is EY1 T classified to the 16SrV‐A subgroup (Lee et al, 1993).…”