A Novel Nested Polymerase Chain Reaction (n-PCR) Assay for Identifying Sorghum nitidum

Shasha Wei,Qin Chen,Zhirui Deng,Renqi Wu,Liping Yin,Jianping Yi

doi:10.15835/nsb346329

Shasha Wei, Qin Chen + Show 4 more

Open Access

https://doi.org/10.15835/nsb346329

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Abstract

This work developed a novel nested polymerase chain reaction (n-PCR) assay to identify Sorghum nitidum (S. nitidum). It has been designed a set of specific n-PCR inner primers Snit5/Snit2 and outer primers Nout1/Nout2 based on a conserved nucleotide sequence of adh1-like gene of S. nitidum. Fourteen samples of sorghum were used to investigate the specificity of the primers and the n-PCR assay. The result showed that 9 samples of S. nitidum displayed a positive strong, specific amplified band at ~873 bp in gel spectra, while other relatives, including Sorghum halepense, Sorghum almum, Sorghum bicolor, Sorghum propinum and Sorghum sudanse exhibited negative amplifications. This assay was able to specifically identify S. nitidum fast and effectively, which could be applied widely in field inspection, agriculture production and plant protection.

Highlights

Sorghum nitidum is a related species of the hard weed Sorghum halepense (Guo et al, 1996), which originated from west of India and spread widely into southeast Asia, Indonesia and Australia
Materials Fourteen sorghum samples were obtained from Shanghai Entry-Exit Inspection and Quarantine Bereau (China) among which 9 samples were S. nitidum, others were S. halepense, S. almum, S. bicolor, S. propinum and S. sudanse, respectively (Tab. 1)
Establishment of typical-PCR assay for identifying sorghum nitidum Primers of typical PCR Snit5/Snit2 were used to amplify the genomic DNA extracted from a single seed of S. nitidum

Summary

Introduction

Sorghum nitidum is a related species of the hard weed Sorghum halepense (Guo et al, 1996), which originated from west of India and spread widely into southeast Asia, Indonesia and Australia. Sorghum Moench could be classified into five groups, namely Stiposorghum, Parasorghum, Eusorghum, Heterosorghum and Chaetosorghum (Celarier, 1959; DeWet, 1978), while S. nitidum was recognized to be one of the most widely spreading species in the group of Parasorghum (Snowden, 1955). The polymorphism of the adh gene could help discover the genetic relationship between different species and provide biological dates for identifying some specific species. The basic principle of n-PCR based on two pairs of specific primers designed to amplify one gene fragment by two round thermo-cycling. We designed a set of specific nPCR primers based on a conserved nucleotide gene fragment of adh1-like gene of S. nitidum and developed a fast assay to identify the S. nitidum with high specificity and effectivity

Materials and methods

Results and discussion

Conclusions