Muskmelon genetic transformation method based on glufosinate-ammonium as selection marker

Abstract

The invention provides a muskmelon genetic transformation method based on glufosinate-ammonium as a selection marker. The method comprises the following steps of: 1) constructing a recombinant expression vector with a BlpR gene, and transferring into agrobacterium to obtain a recombinant strain; 2) infecting the melons cotyledon paraxial end segment by adopting the recombinant agrobacterium in a sterile environment, and placing the infected cotyledon segment on a co-culture medium to grow, so as to obtain an expanded cotyledon explant; 3) transferring the expanded explants to a differential screening culture medium for culture, and inducing cluster buds; 4) separating the elongated cluster buds into single plants, transferring, and further screening and culturing to obtain seedlings with melon overground parts; (5) transferring the seedlings with the melon overground parts into a rooting culture medium, and inducing root differentiation to obtain T0-generation muskmelon transgenic seedlings; and 6) doubly identifying the T0-generation muskmelon transgenic seedlings through glufosinate-ammonium test paper and PCR (Polymerase Chain Reaction) to obtain strains completing genetic transformation. By the method, stably-transformed muskmelon seedlings can be obtained.

Description

technical field [0001] The invention belongs to the technical field of plant molecular genetic breeding and genetic engineering, and relates to a method for efficient genetic transformation of melon and identification of transgenic plants, in particular to a method for genetic transformation of melon based on glufosinate-ammonium as a screening marker. Background technique [0002] Melon (Cucumis melo L.) is an annual herbaceous plant belonging to Cucurbitaceae (Cucumis) and one of the important horticultural crops. Due to the narrow genetic basis of Melon crops, the cultivation of new varieties by traditional breeding techniques is limited to the isolation of intraspecific and interspecific hybridization, and many important traits cannot be improved by conventional breeding methods. In addition, traditional breeding techniques are time-consuming and inefficient. In contrast, transgenic technology has the advantages of high efficiency and precision, and is gradually being u...

AI Technical Summary

Problems solved by technology

However, this method uses PCR technology in the identification of transgenic plants, which requires a large workload, time-consuming and laborious; and the improvement of transformation efficiency is limited.

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