PCR-RFLP identification method for different subgroups of sugarcane white leaf disease phytoplasma

Abstract

The invention relates to a PCR-RFLP identification method for different subgroups of sugarcane white leaf disease phytoplasma. A sugarcane white leaf disease phytoplasma tuf gene is taken as an objectto design a specific primer, a tuf gene fragment is obtained through PCR amplification, then restriction endonuclease Msp I is adopted for digesting the tuf gene fragment obtained through amplification, and the different subgroups of the sugarcane white leaf disease phytoplasma are distinguished through a differentiated RFLP restriction map. The PCR-RFLP identification method for the different subgroups of the sugarcane white leaf disease phytoplasma has the advantages that a sugarcane white leaf disease phytoplasma 16SrXI-B subgroup and a sugarcane white leaf disease phytoplasma 16SrXI-D subgroup can be quickly, accurately and efficiently identified without sequencing, and is important for the pathogen identification and prevention of sugarcane white leaf diseases.

Description

technical field [0001] The invention belongs to the technical field of plant protection, and in particular relates to a PCR-RFLP identification method for sugarcane white leaf disease phytoplasma 16SrXI-B subgroup and 16SrXI-D subgroup. Background technique [0002] Sugarcane white leaf disease is a devastating disease of sugarcane caused by phytoplasma, which can cause huge economic losses to sugarcane and sugar industry. The disease was first discovered in Thailand in 1954, and has now widely occurred in sugarcane-growing countries such as India, Pakistan, Sri Lanka, Laos, Myanmar, Vietnam, the Philippines and Australia. In 2012, the sugarcane white leaf disease phytoplasma was detected for the first time in the Baoshan sugarcane area of ​​Yunnan, my country. Since then, the sugarcane white leaf disease has spread rapidly in the Yunnan sugarcane area, and has now expanded to the Lincang and Pu'er sugarcane areas, posing a serious threat to the sugarcane industry in Yunnan....

AI Technical Summary

Problems solved by technology

[0003] Currently, the distinction between 16SrXI-B and 16SrXI-D subgroups is performed by nested PCR using universal primers (P1 / P7 and R16F2n / R16R2) designed for the phytoplasma 16S rRNA gene, followed by virtual RFLP for nested PCR sequencing. Identification by enzyme digestion analysis, this method requires sequencing and analysis of the product, which is time-consuming and laborious. In addition, errors generated during the sequencing process will also affect the accuracy of the virtual RFLP enzyme digestion analysis results

No method has yet been established for the identification of phytoplasma 16SrXI-B and 16SrXI-D subgroups using actual enzyme digestion

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