Zhang Yong-Jie, Hou Jun-Xiu, Zhang Shu, Hausner Georg, Liu Xing-Zhong, Li Wen-Jia
School of Life Sciences, Shanxi University, Taiyuan, 030006, China.
Institute of Applied Chemistry, Shanxi University, Taiyuan, 030006, China.
Appl Microbiol Biotechnol. 2016 Apr;100(8):3599-610. doi: 10.1007/s00253-016-7287-0. Epub 2016 Jan 12.
Repetitive DNA sequences make up a significant portion of all genomes and may occur in intergenic, regulatory, coding, or even intronic regions. Partial sequences of a serine protease gene csp1 was previously used as a population genetic marker of the Chinese caterpillar fungus Ophiocordyceps sinensis, but its first intron region was excluded due to ambiguous alignment. Here in this study, we report the presence of a minisatellite OsMin1 within this intron, where a 20(19)-bp repeat motif is duplicated two to six times in different isolates. Fourteen intron alleles and 13 OsMin1 alleles were identified among 125 O. sinensis samples distributed broadly on the Tibetan Plateau. Two OsMin1 alleles were prevalent, corresponding to either two or five repeats of the core sequence motif. OsMin1 appears to be a single locus marker in the O. sinensis genome, but its origin is undetermined. Abundant recombination signals were detected between upstream and downstream flanking regions of OsMin1, suggesting that OsMin1 mutate by unequal crossing over. Geographic distribution, fungal phylogeny, and host insect phylogeny all significantly affected intron distribution patterns but with the greatest influence noted for fungal genotypes and the least for geography. As far as we know, OsMin1 is the first minisatellite found in O. sinensis and the second found in fungal introns. OsMin1 may be useful in designing an efficient protocol to discriminate authentic O. sinensis from counterfeits.
重复DNA序列在所有基因组中占很大一部分,可能出现在基因间、调控、编码甚至内含子区域。丝氨酸蛋白酶基因csp1的部分序列先前被用作中华虫草菌的群体遗传标记,但由于比对不明确,其第一个内含子区域被排除在外。在本研究中,我们报告了该内含子中存在一个小卫星OsMin1,在不同分离株中,一个20(19)bp的重复基序重复了两到六次。在广泛分布于青藏高原的125个中华虫草样本中,鉴定出了14个内含子等位基因和13个OsMin1等位基因。两个OsMin1等位基因占主导地位,分别对应于核心序列基序的两次或五次重复。OsMin1似乎是中华虫草基因组中的一个单基因座标记,但其起源尚不确定。在OsMin1的上游和下游侧翼区域检测到丰富的重组信号,表明OsMin1通过不等交换发生突变。地理分布、真菌系统发育和寄主昆虫系统发育均显著影响内含子分布模式,但对真菌基因型的影响最大,对地理因素的影响最小。据我们所知,OsMin1是在中华虫草中发现的第一个小卫星,也是在真菌内含子中发现的第二个小卫星。OsMin1可能有助于设计一种有效的方案,以区分正品中华虫草和假冒品。