A novel multiplex PCR assay for species identification in the Chinese Egret (Egretta eulophotes) and Little Egret (E. garzetta)

Huang, Xunhe; Zhou, Xiaoping; Lin, Qihang; Peng, Zhenni; Fang, Wenzhen; Chen, Xiaolin

doi:10.1007/s12686-011-9466-8

Cited by 5 publications

(5 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although there is still a risk of miss-identifying the species when we use only one SNP as a marker, the multiplex PCR marker presented here distinguished between AAGG from AABB species with complete accuracy. Multiplex PCR is rapid and affordable, allowing simultaneous detection of multiple loci, and thus, has been applied in various species, such as grapevine (Merdinoglu et al , 2005; Migliaro et al , 2013), Chinese egret (Huang et al , 2012) and for the typing of high molecular weight alleles in wheat (Ma et al , 2003).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Multiplex PCR effectively identifies tetraploid Triticum AABB – or AAGG-genome species

Tanno

Takeuchi

Akahori

et al. 2017

Plant Genet. Resour.

View full text Add to dashboard Cite

We developed a multiplex PCR DNA marker for quick and easy identification of the AAGG-genome timopheevii lineage, including Triticum timopheevii, Triticum araraticum and hexaploid Triticum zhukovskyi (AAAmAmGG), and the AABB-genome emmer wheat lineage, including Triticum durum, Triticum dicoccum and Triticum dicoccoides. Distinguishing between tetraploid AAGG- and AABB-genome wheat species based on morphology is known to be difficult. This multiplex PCR system is based on the simultaneous PCR amplification of two chloroplast regions, matK and rbcL. The matK region molecularly distinguishes the two lineages, whereas the rbcL region is a positive control amplicon. We also examined whether the simple sequence repeat is a fixed mutation within species, using genetic resources in the collection of KOMUGI, Kyoto University, which comprises accessioned species collected across diverse geographical areas. The multiplex PCR marker distinguished AAGG from AABB species with complete accuracy.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The upper bands represent rbcL, which is a positive control for confirming the validity of the experiment. 2013), Chinese egret (Huang et al, 2012) and for the typing of high molecular weight alleles in wheat (Ma et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

Multiplex PCR effectively identifies tetraploid Triticum AABB – or AAGG-genome species

Tanno

Takeuchi

Akahori

et al. 2017

Plant Genet. Resour.

View full text Add to dashboard Cite

show abstract

“…In order to test for PCR success rates, DNA samples were amplified for species identification using E. eulophotes-specific ND6 and tRNA Glu gene primers (Huang et al 2012a), which respectively amplified 101 and 278 bp fragments of the mitochondrial DNA (mtDNA) control region. PCR amplifications were performed in 20 μL reactions containing 10 μL Premix Taq (Takara), 0.4 μM of each primer and 50 ng of genomic DNA.…”

Section: Pcr Amplificationsmentioning

confidence: 99%

“…In recent years, many studies have focused on the conservation genetics and molecular ecology of this egret species. These include evaluation of genetic diversity and population structure (Zhou et al 2010), complete mitochondrial genome (Zhou et al 2014), polymorphism and selection of major complex histocompatibility (MHC) genes (Wang et al 2013), species identification (Huang et al 2012a(Huang et al , 2013, sex identification (Wang et al 2011;Huang et al 2012b), isolation of polymorphic microsatellite loci (Huang et al 2010;Dai et al 2013) and primer pairs for amplifying the complete mitochondrial DNA (Zhou et al 2008). However, many evolutionary and ecological questions regarding mating systems and kinship in this egret species remain unclear because of the lack of available genetic information.…”

Section: Introductionmentioning

confidence: 99%

Noninvasive and nondestructive sampling for avian microsatellite genotyping: a case study on the vulnerable Chinese Egret (Egretta eulophotes)

et al. 2015

Self Cite

View full text Add to dashboard Cite

Background: Noninvasive and nondestructive DNA sampling techniques are becoming more important in genetic studies because they can provide genetic material from wild animals with less or even without disturbance, which is particularly useful for the study of endangered species, i.e., birds. However, nondestructively and noninvasively sampled DNA may, in some cases, be inadequate in the amount and quality of the material collected, which can lead to low amplification success rates and high genotyping errors. Methods:In this study, noninvasive (eggshell swab, shed feather and feces), nondestructive (plucked feather and buccal swab) and invasive (blood) DNA samples were collected from the vulnerable Chinese Egret (Egretta eulophotes). DNA concentrations, PCR amplification success and microsatellite genotyping errors of different sample types were evaluated and compared to determine whether noninvasive and nondestructive samples performed as well as invasive samples in our experimental procedures. Results:A total of 159 samples were collected in the field. Among the different sample types, the highest DNA concentrations (154.0-385.5 ng/μL) were obtained from blood. Those extracted from fecal samples were the lowest, ranging from 1.25 to 27.5 ng/μL. Almost all of the DNA samples, i.e., 95.59 %, were successfully amplified for mtDNA (n = 152) and 92.76 % of mtDNA samples were successfully genotyped for at least five of the nine microsatellite loci tested (n = 141). Blood samples and buccal swabs produced reliable genotypes with no genotyping errors, but in feces, allelic dropouts and false alleles occurred in all nine loci, with error rates ranging from 6.67 to 38.10 % for the dropouts and from 6.06 to 15.15 % for the false alleles. Conclusions:These results indicate that both nondestructive and noninvasive samplings are suitable for avian microsatellite genotyping, save for fecal DNA. However, we should remain cautious of the appearance of genotyping errors, especially when using noninvasive material.

show abstract

“…The DNA samples were amplified using species-specific ND6 and tRNA Glu gene primers for species identification (Huang et al 2012) and using Chromo-Helicase-DNA binding protein gene (CHD) primers for sex determination (Wang et al 2011). Then, 13 microsatellite loci isolated from the Chinese Egret (Huang et al 2010;Dai et al 2013) and three markers obtained by cross-amplification from Reddish Egret (Egretta rufescens) (Hill and Green 2011) were chosen and allocated into four multiplex PCR panels using software MPprimer (Shen et al 2010), based on size limitations and annealing temperature of primers.…”

mentioning

confidence: 99%

A novel multiplex PCR assay for individual identification in the vulnerable Chinese Egret (Egretta eulophotes) using molted feathers

Dai

Fang

Lin

et al. 2015

Conservation Genet Resour

Self Cite

View full text Add to dashboard Cite

Noninvasive DNA may be used in individual identification for counting or tracking animals. Here a multiplex PCR assay, in which 16 highly polymorphic microsatellite markers were chosen and allocated into four multiplex PCR panels, was described for the individual identification of the Chinese Egret (Egretta eulophotes) using molted feathers. In this study, the combined exclusion probability for unrelated individuals (P ID ) and for individuals with sibling relationship (P ID(sib) ) were 7.02 9 10 -19 and 2.9 9 10 -7 , respectively. Molted feathers (n = 59) of E. eulophotes amplified by more than 7 loci were unambiguously identified as 43 different individuals (32 females and 11 males). The results indicated that our newly developed assay could be used for noninvasive individual identification in the vulnerable Chinese Egret and other Ardeids.

show abstract

A novel multiplex PCR assay for species identification in the Chinese Egret (Egretta eulophotes) and Little Egret (E. garzetta)

Cited by 5 publications

References 8 publications

Multiplex PCR effectively identifies tetraploid Triticum AABB – or AAGG-genome species

Multiplex PCR effectively identifies tetraploid Triticum AABB – or AAGG-genome species

Noninvasive and nondestructive sampling for avian microsatellite genotyping: a case study on the vulnerable Chinese Egret (Egretta eulophotes)

A novel multiplex PCR assay for individual identification in the vulnerable Chinese Egret (Egretta eulophotes) using molted feathers

Contact Info

Product

Resources

About