Development of loop-mediated isothermal amplification (LAMP)-based SNP markers for shelf-life in melon (Cucumis melo L.).
In this study, LAMP markers linked to shelf-life in melon (Cucumis melo L.) were developed by converting a cleaved amplified polymorphic sequences (CAPS) marker (C2). The CAPS-PCR fragments from the long-shelf-life melon (O-3) and short-shelf-life melon (Nat-2) were cloned and sequenced to construct LAMP primers. A single nucleotide polymorphism (SNP) was identified between O-3 and Nat-2. LAMP primers were designed to detect the SNP. In the LAMP reaction to detect long-shelf-life melon, the turbidity of the templates using O-3, F1, homozygous long-shelf-life F2 lines and heterozygous long-shelf-life F2 lines started to increase after 40 min. In contrast, the turbidity of Nat-2 and homozygous short-shelf-life F2 lines did not increase even after 90 min. In the LAMP reaction to detect short-shelf-life melon, the turbidity of the templates using Nat-2, F1, homozygous short-shelf-life F2 lines and heterozygous long-shelf-life F2 lines started to increase after 40 min. But the turbidity of O-3 and homozygous long-shelf-life F2 lines did not increase after 90 min. This attests to the high reliability and usefulness of LAMP for marker-assisted selection.
Fukuta
Shiro
S
Biotechnology Group, Aichi Pref. Agricultural Research Center, Sagamine. Yazako, Nagakute, Aichi, Japan. shirou_fukuta@pref.aichi.lg.jp
Mizukami
Yuko
Y
Ishida
Akira
A
Kanbe
Michio
M
D001483:Base Sequence; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D028463:Cucumis melo / Q000235:genetics* / Q000378:metabolism; D005524:Food Technology; D005819:Genetic Markers; D006579:Heterozygote; D006720:Homozygote; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D005819:Genetic Markers