Academia Sinica

RAPD analysis of mtDNAs from multiplasmic cms lines
--Wang, Z; Wang, B and Zeng, M

The mitochondrial DNAs of three maize multiplasmic cms (cytoplasmic male sterility) lines: Mo17-cms-19A (T group), Mo17-cms-shang26 (S group), Mo17-cms-C (C group), and their maintainer line Mo17, and a new inbred, Tai-A cms line (unknown group) and its maintainer line C103 were analyzed with 210 random decamer nucleotide primers. The experimental results were as follows:

1) A genetic relationship dendrogram was made and the genetic distance was calculated by cluster analysis with the amplification products clearly amplified from 40 Operon primers. It was found that the relative genetic gap between Tai-A and the other five lines is as long as 24.9, however, its affinity between Mo17-cms-19A and Mo17-cms-C, and the genetic distance, is only 6 between them.

2) A RAPD-fingerprinting map of the six lines was made with five primers: OPAC-02, OPAN-05, OPG-19, OPT-09 and OPT-12, which would provide a rapid and convenient molecular tool to detect the six lines. In the map, RAPD-PCR products: OPAC-02(680), OPAC-02(1053), OPAN-05(680), OPAN-05(370), OPT-12(1230), OPT-09(800) in company with OPG-19(290) can be used as molecular markers to separate and identify Mo17-cms-shang26, Mo17-cms-C, Mo17-cms-19A, Tai-A, Mo17 and C103, respectively.

The mitochondrial DNAs of six lines were digested by restriction endonuclease PstI. From the digestion pattern, it was found that Tai-A apparently differed from the other lines. According to the results of restriction endonuclease pattern and RAPD analysis, we think that Tai-A may be a new kind of cms line differing from T, C, S types and the origin of Mo17-cms-19A, Mo17-cms-shang26 and Mo17-cms-C.

In addition, primer OPZ-05 and primer OPT-09 can amplify special polymorphic products in four cms lines and two maintainer lines respectively. The special RAPD PCR products of OPT-09(800) in C103 and Mo17 were cloned into PUC19 and Southern hybridization results showed that the cloned fragments were either single-copy or low-copy number (Fig. 1).

Figure 1. Amplified polymorphic products for primer OPH-07. 1) Mo17-cms-C; 2) Mo17-cms-19A; 3) Mo17-cms-shang26; 4) TaiA; 5) C103; 6) Mo17. 

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