T/JSF T/JSF001-2024 Technical Specifications for Gender Identification of American Black Willow at the Seedling Stage—SSR Molecular Marker Method

Scope

Scope: This document establishes the procedure for identifying American black poplar seedlings using SSR molecular markers. This document is applicable to the identification of American black poplar seedlings based on their young tissues (leaves, buds, etc.) during the seedling stage, using SSR molecular markers. It covers the principles, main instruments, reagents, operating procedures, and identification conclusions for molecular gender identification of American black poplar seedlings. Main technical content: This document focuses on the impact of poplar pollen dispersion on the industry and establishes a standard system for gender identification of poplar seedlings. This not only accelerates the breeding of non-flying pollen male elite varieties, but also helps reduce the circulation of female commercial seedlings, providing a stable and feasible technical standard for controlling poplar pollen dispersion. The technical content can be generally divided into four parts: DNA extraction, PCR amplification, electrophoretic detection of amplification products, and determination of gender results. 1) DNA extraction: The standard specifies the detailed steps of DNA extraction, including sample grinding, DNA extraction, and DNA quality detection, to ensure high-quality DNA for subsequent gender identification. 2) PCR amplification: Select any pair of SSR primers listed in the standard for gender identification and complete PCR amplification according to the standard-specified reaction system and amplification procedures. 3) PCR amplification product detection: Conduct amplification product detection according to the standard-specified steps to obtain a gel electrophoresis image. 4) Determination of gender results: In the gel electrophoresis image, if the tested sample amplifies to an electrophoretic band of the target fragment size specified in the standard, it is a male plant; otherwise, it is a female plant.

Development Information

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