A PCR-based marker system monitoring CMS-(S), CMS-(T) and (N)-cytoplasm in the onion (Allium cepa L.)

Abstract

The chimerical mitochondrial CMS(1)-specific sequence in chives ( Allium schoenoprasum) was used to develop a PCR-marker that distinguishes both male-sterility inducing cytoplasms, CMS-(S) and CMS-(T), from the normal cytoplasm in onion ( Allium cepa). In combination with a previously described marker for CMS-(S), which anchors in the upstream region of the mitochondrial gene cob, all of the three known cytoplasms in the onion are distinguishable. The PCR-marker system was tested in 361 onion plants, which were selected from F(1)-hybrids and different open-pollinated varieties. The latter are mainly landraces from Turkey, in which all three cytoplasm types were detected.

Testing the utility of matK and ITS DNA regions for discrimination of Allium species

“…This accession was obtained from a local market in Bursa, Turkey, and could be a hybrid onion cultivar with sterile cytoplasm. Indeed, PCR analyses with 2 different PCR-based markers developed by Sato (1998) and Engelke et al (2003) demonstrated that this A. cepa accession had "S" cytoplasm (data not presented). In addition, these markers were also amplified within the genome of A. roylei and the sizes of the PCR-amplified DNA fragment were similar to that of "S" cytoplasm in A. cepa.…”

Inheritance mode of male sterility in bunching onion (Allium fistulosum L.) accessions

“…Since the fertility restoring gene for Zhangqiu-and Guangzhou-derived CMS has not been discovered in this study, we have compared the nucleotide sequences of three mitochondrial genes, cob, coxI and V7 region of srRNA among 'Zhangqiu', 'Guangzhou' and four F 1 commercial cultivars, 'Natsuougi-2', 'Fuyuougi-2', 'Ryuushou' and 'White Tower'. The primer sequences and PCR conditions for amplification of these genes were according to the reports of Engelke et al (2003) for cob, Kim et al (2009) for coxI and Buiteveld et al (1998) for V7 region of srRNA. In the three regions, sequence polymorphism was not detected among the six materials (data not shown).…”

The Application of Molecular Markers to Accelerate the Recovery of Cytoplasmic and Nuclear Male Sterility in South African Onion (Allium cepa L.) Hybrid Parental Lines

“…The orfA501 marker (Engelke et al, 2003) successfully amplified a 473bp chimerical orfA501 fragment associated with S or T cytoplasm (another system of male sterility not commonly used in breeding) in all onion samples designated as S (Figure 1; orfA501; line 1: all samples except 5 and 7; line 2: all samples; lines 3 and 4: only samples 6 and 7, respectively). Sample 8 of line 1 did not initially show a fragment with marker orfA501 (Figure 1, cytoplasmic genotype S * ), and was attributed to omitted gDNA template in the PCR.…”

“…A 180bp fragment is amplified from the N-cytoplasm with a N-specific primer and the same common reverse primer. The use of the three primers (S-, N-specific and common reverse primer) together in the same PCR is recommended by Engelke et al (2003) so that sub-stoichiometric amounts of the 414bp S-fragment is not amplified in N-cytoplasm. Both fragments are amplified in S-cytoplasm.…”

“…The orfA501 marker is based on an insertion in the mitochondrial sequence of chives that forms an open reading frame (orfA501) (Engelke et al, 2003). Primers which span nearly the complete orfA501 amplify a 473bp fragment in both T-and S-cytoplasm in onion, but nothing in N-cytoplasm.…”