Ultrastructure of phytoplasma-infected jujube leaves with witches' broom disease

The subcellular characteristics of phytoplasma-infected jujube (Ziziphus jujuba) leaves were investigated using transmission electron microscopy. Midrib fragments of witches' broom-diseased jujube leaves were collected from abnormally small leaves at an early stage of branch clustering. The diseased jujube leaves showed multivesicular bodies (MVBs) with vesicles and tubules in the phloem parenchyma cells and sieve elements. The MVBs were connected to the plasma membrane appressed to the cell wall. There were increased callose collars at the pore-plasmodesma unit ends of the sieve elements in the diseased leaves than in control leaves. The proliferation of MVBs in the diseased jujube leaves could be associated with endoplasmic reticulum stress-dependent exosome release. The phytoplasma produced pleomorphic cells in sieve elements. Several types of putative extracellular structures were observed on the phytoplasma cells: (i) fimbriae-like threads, (ii) pili-like projections, (iii) flagella-like appendages, and (iv) tube-like structures. This study provides novel insights into intracellular obligate cell wall-less prokaryotes and host phloem structures.