Method for identifying consanguinity of saccharum arundinaceum in sugarcane via spread sequence

Abstract

The invention relates to a method for identifying consanguinity of saccharum arundinaceum in sugarcane via a spread sequence. The method comprises steps of preparing a root-tip cell slide, preparing alabeled probe, pre-processing the root-tip slide, preparing a denaturing hybrid mixed solution and hybridizing the denaturing hybrid mixed solution and the pre-processed root-tip slide. A to-be-detected sample is judged that the sample contains the consanguinity of saccharum arundinaceum and is a real hybrid later generation of sugarcane and saccharum arundinaceum when a green fluorescence signalis generated on the hybrid slide. The nucleotide sequence of a probe which generates the green fluorescence signal is shown as SEQ ID NO:1 in a sequence list. The method provided by the invention, through the combination with fluorescence in-situ hybridization technology and by identifying the real hybrid later generation of sugarcane and saccharum arundinaceum through the spread sequence probe that saccharum arundinaceum is specific to sugarcane, has the characteristics of being high in efficiency, good in sensitivity, good in stability and high in reliability; and the reality of any hybridgenerations can be detected just by acquiring root tips of sugarcane seedlings, so that an efficient and stable identifying method is provided for hybridization utilization of the saccharum arundinaceum in a sugarcane genetic breeding improving process.

Description

technical field [0001] The invention relates to a method for identifying the lineage of sagegrass in sugarcane, in particular to a method for identifying the lineage of sagegrass in sugarcane by using scattered sequences, and belongs to the technical field of plant detection and identification. Background technique [0002] Sugarcane belongs to annual or perennial tropical and subtropical C 4 Crops, primarily used to make sugar and as a clean energy substitute. Modern sugarcane cultivars are mainly derived from hybrids of a few tropical species and wild species with thin stems, which limits the improvement of sugarcane quality. In order to broaden the genetic basis of sugarcane, sugarcane breeders have introduced wild species of closely related genera such as Pseudomonas spp. into the background of sugarcane by means of distant hybridization, and have obtained a large number of hybrid offspring of sugarcane and Pseudomonas spp. At present, the authenticity identification m...

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Problems solved by technology

Zhuang Nansheng et al. and Wang Ying et al. successively used AFLP marker technology to screen out 3 specific DNA sequences of P. chinensis, and developed them into 3 specific molecular markers of P. chinensis. However, none of them were tested for stability, and it is impossible to know that these markers can be used for Accurate detection of sarcophagus chromosomes or fragments in high-generation clones

In addition, D'hont et al. designed a pair of primers on the 5S rDNA conserved sequence of sugarcane and Banmao by taking advantage of the different lengths of the 5S rDNA non-transcribed spacer NTS sequences between sugarcane and Banmao. According to the electrophoresis detection strip The difference in band size accurately identifies the F 1 However, the 5S rDNA sequence only exists on a certain chromosome of Banmao, which cannot stably and effectively detect the hybrid offspring that lost this chromosome

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