Functional analysis and expression patterns of members of the FLOWERING LOCUS T (FT) gene family in Lilium.

Lilium is an important commercial flowering species, and there are many varieties and more than 100 species of wild Lilium. Lilium × formolongi is usually propagated from seedlings, and the flowering of these plants is driven mainly by the photoperiodic pathway. Most of the other lily plants are propagated via bulblets and need to be vernalized; these plants can be simply divided into pretransplantation types and posttransplantation types according to the time at which the floral transition occurs. We identified three Lilium FLOWERING LOCUS T (LFT) family members in 7 Lilium varieties, and for each gene, the coding sequence of the different varieties was identical. Among these genes, the LFT1 gene of Lilium was most homologous to the AtFT gene, which promotes flowering in Arabidopsis. We analyzed the expression patterns of LFT genes in Lilium × formolongi seedlings and in different Lilium varieties, and the results showed that LFT1 and LFT3 may promote floral induction. Compared with LFT3, LFT1 may have a greater effect on floral induction in Lilium, which is photoperiod sensitive, while LFT3 may play a more important role in the floral transition of lily plants, which have a high requirement for vernalization. LFT2 may be involved in the differentiation of bulblets, which was verified by tissue culture experiments, and LFT1 may have other functions involved in promoting bulblet growth. The functions of LFT genes were verified by the use of transgenic Arabidopsis thaliana plants, which showed that both the LFT1 and LFT3 genes can promote early flowering in Arabidopsis. Compared with LFT3, LFT1 promoted flowering more obviously, and thus, this gene could be an important promoter of floral induction in Lilium.

Evaluation of resistance to Botrytis elliptica in Lilium hybrid cultivars.

Gray mold disease, caused by the fungus Botrytis elliptica, is one of the major diseases affecting Lilium species, and it has become a limiting factor in the production of ornamental lilies. To support selecting and breeding Botrytis-resistant cultivars, a total of 50 Lilium cultivars belonging to seven hybrid types were evaluated using a detached leaf technique for resistance to B. elliptica. Through resistance evaluations, Oriental × Trumpet and Oriental hybrid cultivars were classified as resistant lines, while Asiatic and Trumpet hybrids were classified as susceptible lines. A highly resistant (HR) Oriental hybrid, 'Sorbonne', and a highly susceptible (HS) Asiatic hybrid, 'Tresor', were selected for further study of early host-parasite interactions. After infection, B. elliptica grew faster and more easily on the leaves of 'Tresor' than on those of 'Sorbonne' during initial infection; when 'Tresor' leaves were completely infected, only a few lesions were observed on 'Sorbonne' leaves. Biochemical differences between these two cultivars were found following inoculation with B. elliptica, as shown by studies of reactive oxygen species (ROS) and the enzymatic antioxidant system. Rapid accumulation of H2O2 and ·O2- to trigger a defense response was detected in HR 'Sorbonne'. Meanwhile, higher levels of antioxidant activity, including SOD, POD and CAT, were found in HR 'Sorbonne' than in HS 'Tresor' before 48 h post-inoculation, but antioxidant activity was reduced with subsequent infection progress. These large and timely increases in ROS and antioxidant activities could be the main contributors to the high resistance of the 'Sorbonne' cultivar.

Transcriptome-Wide Analysis of Botrytis elliptica Responsive microRNAs and Their Targets in Lilium Regale Wilson by High-Throughput Sequencing and Degradome Analysis.

MicroRNAs, as master regulators of gene expression, have been widely identified and play crucial roles in plant-pathogen interactions. A fatal pathogen, Botrytis elliptica, causes the serious folia disease of lily, which reduces production because of the high susceptibility of most cultivated species. However, the miRNAs related to Botrytis infection of lily, and the miRNA-mediated gene regulatory networks providing resistance to B. elliptica in lily remain largely unexplored. To systematically dissect B. elliptica-responsive miRNAs and their target genes, three small RNA libraries were constructed from the leaves of Lilium regale, a promising Chinese wild Lilium species, which had been subjected to mock B. elliptica treatment or B. elliptica infection for 6 and 24 h. By high-throughput sequencing, 71 known miRNAs belonging to 47 conserved families and 24 novel miRNA were identified, of which 18 miRNAs were downreguleted and 13 were upregulated in response to B. elliptica. Moreover, based on the lily mRNA transcriptome, 22 targets for 9 known and 1 novel miRNAs were identified by the degradome sequencing approach. Most target genes for elliptica-responsive miRNAs were involved in metabolic processes, few encoding different transcription factors, including ELONGATION FACTOR 1 ALPHA (EF1a) and TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR 2 (TCP2). Furthermore, the expression patterns of a set of elliptica-responsive miRNAs and their targets were validated by quantitative real-time PCR. This study represents the first transcriptome-based analysis of miRNAs responsive to B. elliptica and their targets in lily. The results reveal the possible regulatory roles of miRNAs and their targets in B. elliptica interaction, which will extend our understanding of the mechanisms of this disease in lily.