Development of molecular markers linked to the stability gene of pepper CGMS and its use for determining the types of allele

摘要

A method for determining gene related with stability of a red pepper cytoplasmic male sterility is provided to be able to shorten the time for 2-3 years required for determination and be able to increase accuracy and reliability of the determination because of not being influenced by environment. The method comprises the steps of: (a) making a colony where a red pepper showing stable cytoplasmic male sterility(St^2_) and a red pepper showing stable cytoplasmic male sterility(St^u St^u) are separated by a phenotype; (b) extracting DNA from young red pepper leaves of the colony; (c) investigating an amplified fragment length polymorphism(AFLP) molecular marker linked to the gene related with stability of red pepper cytoplasmic male sterility; (d) preparing a linkage map from the investigated AFLP molecular marker; (e) analyzing a nucleotide sequence of E112M313(SEQ ID : NO. 1) to change the linked AFLP molecular marker E112M313(SEQ ID : NO. 1) into an amplified fragment cut polymorphism(CAPS) molecular marker capable of being utilized to real breeding; (f) determining SEQ ID : NO. 3 around the SEQ ID : NO. 1 bidirectionally; (g) selecting a primer combination of SEQ ID : NO. 4 and SEQ ID : NO. 5 using the SEQ ID : NO. 3; (h) performing PCR on the DNA extracted from the red pepper leaves to be subjected to analysis using the primer combination; and (i) after treating the amplified PCR product with a restriction enzyme of MseI, electrophoresing it.