To catalog and preserve 77 tobacco germplasm resources collected in Hubei Province into the National Bank of Tobacco Germplasm Resources, field identification and evaluation of these tobacco germplasm resources were carried out. Data on period of duration，morphological characteristics, main agronomic traits and resistance to main virus diseases, have been obtained, and the photos of plant type, leaf, inflorescence, corolla and capsule also have been taken individually. Result showed that the germplasms B035, B075 and B076 have resistance or mid-resistance for more than two main virus diseases, while germplasm B025 have immunity to TMV. Field phenotypic characterization and SSR marker analysis were applied to identify the cataloging germplasm, and to remove the repeated collection germplasm resources. The obtained results of this study will lay a good foundation for enriching the the genetic diversity of National Bank of Tobacco Germplasm Resources and the tobacco breeding.