No abstract

In this research we eliminated chrysanthemum stunt viroid (CSVd) from a highly infected chrysanthemum cultivar using a newly established method. 'Piato' is one of the most difficult cultivars in which to obtain CSVd-free plants by conventional methods. Leaf primordium-free shoot apical meristems (LP-free SAMs) of 'Piato' plants were dissected and attached to CSVd-free chrysanthemum or cabbage root tips. As shown by nested-PCR, CSVd was not detected in some shoots regenerated on both types of root tip. The production rates of CSVd-free plants using chrysanthemum and cabbage root tips were 14% and 3%, respectively. Regeneration of plants from LP-free SAMs of chrysanthemum plants by attaching these SAMs to root tips is an efficient method of generating CSVd-free chrysanthemum plants.

Cultivars resistant to chrysanthemum stunt viroid (CSVd) are desirable for stable production of chrysanthemum (Chrysanthemum × morifolium Ramat.). We previously reported that CSVd was absent not only from shoot apical meristems (SAMs) but also from leaf primordia (LP) of resistant plants following CSVd inoculation. Using this characteristic as a phenotypic marker, we could identify several resistant cultivars in this study. CSVd was inoculated directly into the SAMs of 85 commercial cultivars by attaching SAMs to infected root tips in vitro, and the cultivars were tested for the presence of CSVd in newly expanded leaves. Of the 85 cultivars, 20 resistant candidates could be identified. These plants were classified into two types according to their CSVd infection characteristics: a CSVd-uninfected or slow titer-increasing type and a CSVd-disappearance type. Scions of four of the 20 candidate cultivars were grafted to CSVd-infected rootstocks for evaluation of the CSVd titer in newly expanded leaves. Although CSVd was detected at high titer in two candidate cultivars over the two-month testing period, 'Sei no Issei' and 'Mari Kazaguruma', other two candidate cultivars, proved to be resistant cultivars. In 'Sei no Issei', CSVd was detected temporarily after grafting, but titers decreased in the newly expanded leaves, while CSVd was not detected in a shoot tip of an infected plant and only at trace levels in a young leaf. 'Mari Kazaguruma' also proved to be a resistant cultivar with a slow titer increase. The different types of CSVd resistance in chrysanthemum cultivars will contribute to CSVd-resistant breeding. With some modifications, the in vitro screening method established here will be available for broadening the diversity of genetic resources resistant to CSVd.

Chrysanthemum ( Dendranthema grandiflorum Kitam.) is one of the qualitative short-day flowering plants. Therefore, the flowering of chrysanthemum can usually be controlled by photoperiod. However, it was noted that 'Piato' plants infected by the chrysanthemum stunt viroid (CSVd) flowered autonomously even under long-day conditions. In this study, CSVd-free and CSVd-infected plants were prepared by culturing different-sized dissected shoot apical meristems (SAMs) of 'Piato'. Using these CSVd-free and CSVd-infected plants, we clarified the relationship between CSVd infection and the autonomous flowering of 'Piato'. Under natural short-day conditions, the flowering of plants regenerated from SAMs containing leaf primordia (LPs) was 1 month earlier than plants regenerated from LP-free SAMs. CSVd was detected from these early flowering plants by reverse transcription-polymerase chain reaction. On the other hand, CSVd was not detected in plants regenerated from LP-free SAMs. CSVd-infected and CSVd-free plants were grown under long-day conditions simulated by night-break lighting at 22:00 p.m. to 2:00 a.m. All CSVd-infected plants flowered autonomously even under long-day conditions; on the other hand, CSVd-free chrysanthemum plants maintained their vegetative growth. When the CSVd-free plants were inoculated with CSVd by grafting them to CSVd-infected rootstocks, they flowered autonomously even under night-break lighting. In this study, the results suggest that CSVd may control the qualitative development process, flowering, i.e. CSVd can induce the autonomous flowering of chrysanthemum.

No abstract