Diversity and inheritance of inter-simple sequence repeat polymorphisms in Douglas-fir (Pseudotsuga menziesii) and sugi (Cryptomeria japonica)

Abstract

We studied inter-simple sequence repeat (ISSR) polymorphism and inheritance in Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] and sugi (Cryptomeria japonica D. Don) megagametophytes using primers that anneal to simple repeats of various lengths, sequences, and non-repetitive motifs at the 5' and 3' ends. Products were visualized on agarose gels with ethidium bromide staining. More than 60% of the 96 primers tested gave interpretable banding patterns in both Douglas-fir and sugi, and the useful primers were in complete agreement among species. Dinucleotide repeat primers were the majority of those tested, and gave all of the useful banding patterns. The 24 best primers were used for segregation studies, yielding a total of 77 loci distributed among two Douglas-fir families and one sugi family. Approximately 90% of the 24 primers showed polymorphism within at least one of the three families. The average number of variable loci per primer was 1.6. Primers based on (AG) n repeats gave the largest number of polymorphic loci; 16 primer-family combinations yielded 24 segregating loci. However, primer based on (GT) n repeats gave the most loci per primer studied (mean of 2.0). All markers displayed apparent dominance (band presence vs absence), and all but three segregation ratios (4%) fit Mendelian expectations: Because they employ longer primers than do RAPDs, have a high degree of polymorphism, conform well to Mendelian expectations, and do not require use of acrylamide gels for analysis, ISSRs may be useful markers for PCR-based genome maps and population studies of conifers.

Plant Regeneration and Genetic Diversity of Regenerants from Seed-derived Callus of Reed (Phragmites communis Trinius)

“…지금 까지 Prunus dulcis의 RAPD 및 ISSR 분석 (Martins et al, 2004), Malus pumila의 RAPD 분석 (Modgil et al, 2005), Pinus thunbergii의 RAPD 분석 (Goto et al, 1998), 장기간 기 내 배양된 춘란(Cymbidium goeringii)과 한란(C. kanran)의 유전적 변이성 (Ryu et al, 2011 (Fang and Roose, 1997;Tsumura, 1996). 또한 AFLP (amplified fragment length polymorphism)보다 과정이 간단 하고 (Russel et al, 1997), RAPD보다는 primer 당 다형성 밴 드수가 더 많으며 (Iruela et al, 2002), primer들의 높은 결합 온도와 더 긴 서열 때문에 RAPD보다 정확하고 재현성이 우수한 밴드들이 생성되는 장점이 있다 (Esselman et al, 1999), 특히 ISSR이 목표로 하는 반복 서열은 진핵 게놈 전 체에 걸쳐 풍부하고 빠르게 변화하여 유전자 지문의 검출 에서 그 유용성이 입증되고 있다 (Godwin et al, 1997;Russel et al, 1997).…”

Genetic Analyses of Casuarinas Using ISSR and FISSR Markers

“…Genome analysis in woody perennials using ISSR has also been shown to be possible in Douglas fir and Sugi (Tsumura et al, 1996), Citrus Fang et al, , 1998, Cocoa (Charters & Wilkinson, 2000), Larix (Arcade et al, 2000) and Morus (Awasthi et al, 2004). In fingerprinting applications, the molecular method employed should be inexpensive, technically simple yet reliable.…”

“…ISSR-PCR has been used widely in plants for the analysis of genetic relationships between and within species (Salimath et al, 1995;Tsumura, Ohba & Strauss, 1996;Fang, Krueger & Roose, 1998;Ge & Sun, 1999;Ajibade, Weeden & Chite, 2000;Chapman, Parh & Oraguzie, 2000;Huang & Sun, 2000;Martin & Sanchez-Yelamo, 2000), assessment of hybridisation in natural populations (Wolfe, Xiang & Kephart, 1998) and germplasm analysis (Proven, Powell & Waugh, 1996;Gilbert et al, 1999). Further, ISSR-PCR is useful in fingerprinting and characterisation of accessions Moreno, Martin & Ortiz, 1998;Blair, Panaud & McCouch, 1999;Charters & Wilkinson, 2000) and identification of cultivars and varieties Prevost & Wilkinson, 1999;Zavodna et al, 2000;Kumar et al, 2001).…”

Genetic structuring in the Pyramid Elimia, Elimia potosiensis (Gastropoda, Pleuroceridae), with implications for pleurocerid conservation

“…Primers for ISSR amplification are complementary to microsatellite sites, bind to them using a one to three nucleotide anchor sequence on the 5' or 3' end (Bussell et al 2005), and amplify the ISSR in between (Culley and Wolfe 2001). This approach does not require prior genome sequence information, and leads to banding patterns containing multiple loci and high levels of polymorphism (Tsumura et al 1996). ISSRs have provided useful population data in a variety of organisms including plants (Lisek and Rozpara 2010), insects (Vijayan et al 2006), arachnids (Machkour-M'Rabet et al 2009), and birds (Haig et al 2003), but have not been utilized extensively in gastropods (Dong et al 2011, Snegin 2014).…”