The study was conducted at the College of Agricultural Engineering Sciences-Department of Plant Protection in 2018-2019 Baghdad/Iraq. The research aims to molecular identification of the fungal pathogen Neoscytalidium sp. the causal agent of the wilt branches disease on pomegranate, mulberry, apple, castor, and rubber by 18S ribosomal RNA gene and DNA sequencing, to demonstrate the pathogenicity, and host range of the fungal isolates. Five fungal isolates have been recoverd from the infected branches belonged to the five types of trees such as; apple (Malus domestica), pomegranate (Punica granatum), mulberry (Morus alba), india rubber (Ficus elastica), castor (Ricinus communis). The results showed compatibility of 100% with a standard in Gene Bank from Neoscytalidium novaehollandiae in pomegranate, mulberry, apple, and rubber, while in Castor having nine Trinsvertion C>G, A>T, T>G, T>G, G>C, T>G, A>C, A>T, and T>A, and five Transition G>A, G>A, A>G, A>G, and G>A) having 97% compatibility with standard in Gene Bank .This is the first record of the pathogen , N. novaehollandiae on these host trees in Iraq. The pathogenicity test confirmed their ability to infected all types of trees with different symptoms were observed, the results also, showed difference in infection severity which was found to be higher on the apple trees(88.09%).This result indicates the more susceptibility of apple trees to fangal sooty stem disease compared to other trees.