A novel method of organogenesis in neem (Azadirachta indica A. Juss.) from unfertilized ovaries is described. The Murashige and Skoog's (MS) medium with 9 % sucrose, 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 5 μM 6-benzylaminopurine (BAP) was the best for callus induction from unfertilized ovaries. However, further proliferation of callus occurred better on MS medium supplemented with 0.5 μM 2,4-D either alone or in combination with 4.5 μM kinetin. Maximum shoot regeneration (78 %) was observed when calli, induced from ovaries of 4 mm size flower buds and proliferating on MS + 0.5 μM 2,4-D, were subcultured to MS medium containing 5 μM BAP. Histological analysis revealed that 4 mm sized flower bud corresponds to a 2-nucleate stage of embryo sac. The shoots were then multiplied by forced axillary branching on MS medium supplemented with 1.0 μM BAP and 250 mg dm -3 casein hydrolysate. The shoots could be rooted on ¼ strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA) at a frequency of 79 %. Cytological analysis by root tip squash preparations revealed that all the plantlets were diploids. These plants were subsequently hardened and established in soil with transplantation rate of 81.8 %.Additional key words: adventitious shoot proliferation, auxins, callus culture, casein hydrolysate, cytokinins, embryo sac, histology, ovary culture. ⎯⎯⎯⎯Neem (Azadirachta indica A. Juss.), an evergreen tropical tree belonging to the family Meliaceae, has many important medicinal, agrochemical and economic uses. Due to highly heterozygous nature, long reproductive cycle and poor seed yield, improvement of neem by conventional methods is very limited. In this respect, tissue culture can play an important role. In vitro regeneration of neem from various vegetative tissues was published (Chaturvedi et al. 2004a, Rout 2005. Recently, some studies have described shoot regeneration from anthers/microspores (Chaturvedi et al. 2003a) and endosperm tissues (Chaturvedi et al. 2003b) of adult tree origin.Even though the main objective of the present study was to obtain haploid plants from the unfertilized ovary, regeneration of diploid plants indicated the potential use of this juvenile unfertilized ovary explant for large scale micropropagation. In contrast, the leaves, nodal and internodal segments from mature trees show recalcitrance due to accumulation of secondary metabolites.Unfertilized ovaries, obtained from closed flower buds of an adult 54-year-old neem tree, were used as explants. Ovaries were excised from flower buds of four sizes (2, 3, 4 and 5 mm) and the corresponding developmental stage of ovary was determined by paraffin sections. The flower buds were surface sterilized with 0.1 % (m/v) solution HgCl 2 for 7 min, followed by three washings in sterile-distilled-water. The ovaries were dissected out with the aid of a stereo-microscope (Nikon SMZ-645, Tokyo, Japan). Four ovaries were cultured in ⎯⎯⎯⎯

The human leukocyte antigen (HLA) class II proteins present peptides to CD4+ T cells through an interaction with T cell receptors (TCRs). Thus, HLA proteins are key players in shaping immunogenicity and immunodominance. Nevertheless, factors governing peptide presentation by HLA-II proteins are still poorly understood. To address this problem, we profiled the blood transcriptome and immunopeptidome of 20 healthy individuals and integrated the profiles with publicly available immunopeptidomics datasets. In depth multi-omics analysis identified expression levels and subcellular locations as import sequence-independent features governing presentation. Levering this knowledge, we developed the Peptide Immune Annotator Multimodal (PIA-M) tool, as a novel pan multimodal transformer-based framework that utilises sequence-dependent along with sequence-independent features to model presentation by HLA-II proteins. PIA-M illustrated a consistently superior performance relative to existing tools across two independent test datasets (area under the curve: 0.93 vs. 0.84 and 0.95 vs. 0.86), respectively. Besides achieving a higher predictive accuracy, PIA-M with its Rust-based pre-processing engine, had significantly shorter runtimes. PIA-M is freely available with a permissive licence as a standalone pipeline and as a webserver (https://hybridcomputing.ikmb.uni-kiel.de/pia). In conclusion, PIA-M enables a new state-of-the-art accuracy in predicting peptide presentation by HLA-II proteins in vivo.