Large-Scale In Vitro Multiplication and Phytochemical Analysis of Himantoglossum affine (Boiss.) Schltr.: An Endangered Euro-Mediterranean Terrestrial Orchid

Himantoglossum affine is a threatened terrestrial orchid. We aimed to optimize asymbiotic seed germination and direct embryogenesis analyze the phytochemical profile physico-biochemical analysis of leaf tuber. The individual use organic nitrogen compounds resulted in higher efficiencies, while shortest times were observed using coconut water plus casein hydrolysate. Plantlets grown on media supplemented with pineapple juice peptone had highest plantlet length weight. For embryogenesis, regeneration rate (44%) embryo number/explant (10.12 ± 2.08) young protocorm-like body (PLB) explants 0.5 mg/L naphthalene acetic acid (NAA) 1 thidiazuron (TDZ). During acclimatization process, scattered vascular tubes converted fully developed tissues, ensuring maximum sap flux. Gas chromatography–mass spectrometry identified 1,2,3-propanetriol, monoacetate, 4H-pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl, 2-butenedioic acid, 2-methyl-, (E)- as most prevalent compounds. reported contents total phenolics flavonoids antioxidant activity compared other orchids. glucomannan content (36.96%) was also than starch (31.31%), comparable those tuberous Based fragmentation H. populations Middle East Euro-Mediterranean countries due over-harvesting, climate change, and/or human impact, our procedure offers tool for re-introduction vitro-raised plants areas.