Cloning, Sequence Analysis And PCR Markers Development Of 75K γ-secalin Genes From Secalin Cereale (rye)

Secale L, one of the relative genera of Triticum, has been playing an important tertiary gene pool in wheat breeding for its superior traits in many fields, such as multiple tillers, supernumerary spikelets, high content of lysine and good tolerance to diseases, pests, drought and cold environment. The wheat-rye 1BL/1RS translocation lines, derived from the introgression of rye 1RS into wheat 1BL, have made great contributions in wheat breeding.The rye chromosome arms of 1RS, 1RL and 2RS carry some genes which control the quality of storage protein. And the coding products of secalin genes in 1RS have negative effects on the baking quality of dough. The 75K 7-secalin, coded by the corresponded genes on 2RS, accounts for half contents of the total seed protein, however we still are lack of awareness of both the features of the coding genes and the role of its expression products.In order to investigate the genetic diversity and analysis the sequence features of the 75K 7-secalin gene family, three ryes (Secale cereale L.) were studied by SDS-PAGE and molecular cloning. The results are as following: 1. Cloning and sequencing of the coding sequences of 75K γ-secalinWe designed a pair of special primers based on the known sequence of 75K 7-secalin in GenBank. The rye cultivars Clse 20, Clse 35 and PI 168199 were respectively amplified a 1.1 kb c.a. product using this primer pairs. Cloned these fragments to T-Vector and the positive clones of Clse 20, Clse 35 and PI168199, were named as pSec-1, pSec-2 and pSec-3, respectively. The sequencing results showed that the real length of the three clones were 1194 bp, 1230 bp and 1017 bp, respectively. The predicted amino-acid sequences were 397 aa, 409 aa and 338 aa, deduced by DNAMAN software. 2. Genetic information of the 75K γ-secalin...