Ma Xuhui, Wang Qing, Wang Yanzhi, Ma Jieyun, Wu Nan, Ni Shuang, Luo Tengxiao, Zhuang Lifang, Chu Chenggen, Cho Seong-Woo, Tsujimoto Hisashi, Qi Zengjun
a State Key Laboratory of Crop Genetics and Germplasm Enhancement, JCIC-MCP, Nanjing Agricultural University, Nanjing 210095, China.
b Monsanto Company, 21120 Hwy 30, Filer, ID 83328, USA.
Genome. 2016 Jul;59(7):485-92. doi: 10.1139/gen-2016-0047. Epub 2016 May 11.
Chromosome engineering is an important approach for generating wheat germplasm. Efficient development of chromosome aberrations will facilitate the introgression and application of alien genes in wheat. In this study, zebularine, a DNA methylation transferase inhibitor, was successfully used to induce chromosome aberrations in the octoploid triticale cultivar Jinghui#1. Dry seeds were soaked in zebularine solutions (250, 500, and 750 μmol/L) for 24 h, and the 500 μmol/L treatment was tested in three additional treatment times, i.e., 12, 36, and 48 h. All treatments induced aberrations involving wheat and rye chromosomes. Of the 920 cells observed in 67 M1 plants, 340 (37.0%) carried 817 aberrations with an average of 0.89 aberrations per cell (range: 0-12). The aberrations included probable deletions, telosomes and acentric fragments (49.0%), large segmental translocations (28.9%), small segmental translocations (17.1%), intercalary translocations (2.6%), long chromosomes that could carry more than one centromere (2.0%), and ring chromosomes (0.5%). Of 510 M2 plants analyzed, 110 (21.6%) were found to carry stable aberrations. Such aberrations included 79 with varied rye chromosome numbers, 7 with wheat and rye chromosome translocations, 15 with possible rye telosomes/deletions, and 9 with complex aberrations involving variation in rye chromosome number and wheat-rye translocations. These indicated that aberrations induced by zebularine can be steadily transmitted, suggesting that zebularine is a new efficient agent for chromosome manipulation.
染色体工程是创制小麦种质的重要方法。高效诱导染色体畸变将有助于外源基因导入小麦并在小麦中应用。本研究中,DNA甲基转移酶抑制剂zebularine成功用于诱导八倍体小黑麦品种京辉1号的染色体畸变。将干种子浸泡在zebularine溶液(250、500和750 μmol/L)中24小时,对500 μmol/L处理进行了另外三个处理时间的测试,即12、36和48小时。所有处理均诱导了涉及小麦和黑麦染色体的畸变。在67株M1植株中观察到的920个细胞中,340个(37.0%)携带817个畸变,平均每个细胞0.89个畸变(范围:0 - 12)。这些畸变包括可能的缺失、端体和无着丝粒片段(49.0%)、大片段易位(28.9%)、小片段易位(17.1%)、中间易位(2.6%)、可携带多个着丝粒的长染色体(2.0%)和环状染色体(0.5%)。在分析的510株M2植株中,发现110株(21.6%)携带稳定的畸变。这些畸变包括79株黑麦染色体数目不同的植株、7株小麦和黑麦染色体易位的植株、15株可能存在黑麦端体/缺失的植株以及9株涉及黑麦染色体数目变异和小麦 - 黑麦易位的复杂畸变植株。这些结果表明zebularine诱导的畸变能够稳定遗传,这表明zebularine是一种新的高效染色体操作试剂。