Biochemistry Department, Rothamsted Experimental Station, AL5 2JQ, Harpenden, Herts., U.K..
Plant Mol Biol. 1985 Jul;5(4):213-22. doi: 10.1007/BF00020639.
In potato line Mb1501B one or possibly two normal size Ti TL-DNA copies per tetraploid genome were detected by Southern blot analysis, but no TR-DNA. The TL-DNA probably contained the entire transposon Tn1831 inserted at the T-DNA auxin gene for transcript 2. Northern blot analyses of the steady-state RNA in different Mb1501B tissues isolated from (i) shoots cultured in vitro (ii) grafted plants and (iii) tubers, showed that that TL-DNA transcripts 3, 4, 6a and 7 were expressed most abundantly in the cultured shoots. They formed approximately 0.0023 to 0.0007% of the total poly(A) RNA. Transcripts 1, 5 and 6b were not detected in any of the tissues analysed. This indicated even lower levels of expression (below approximately 0.0001% of the total poly(A) RNA or, making certain assumptions, an abundance of less than one T-DNA derived RNA molecule per cell). As expected, transcript 2 was not detected in any of the Mb1501B tissues. The abundance of the transcripts was reduced in grafted plants and tubers compared with cultured shoots with the greatest decrease (5×) for transcripts 4, 6a and 7. Transcript 4, the one most responsible for the changed growth and development of Mb 1501 B, formed approximately 0.0003% of the poly(A) RNA from both grafted plants and tubers.
在马铃薯品系 Mb1501B 中,通过 Southern 印迹分析检测到每个四倍体基因组中有一个或两个正常大小的 Ti TL-DNA 拷贝,但没有 TR-DNA。TL-DNA 可能包含完整的转座子 Tn1831,插入到 T-DNA 生长素基因中用于转录本 2。来自不同 Mb1501B 组织的(i)体外培养的芽、(ii)嫁接植株和(iii)块茎的稳定态 RNA 的 Northern 印迹分析表明,TL-DNA 转录本 3、4、6a 和 7 在培养的芽中表达最丰富。它们约占总 poly(A) RNA 的 0.0023%至 0.0007%。在任何分析的组织中都没有检测到转录本 1、5 和 6b。这表明表达水平更低(低于总 poly(A) RNA 的约 0.0001%,或者根据某些假设,每个细胞中少于一个 T-DNA 衍生的 RNA 分子的丰度)。正如预期的那样,在任何 Mb1501B 组织中都没有检测到转录本 2。与培养的芽相比,嫁接植株和块茎中的转录本丰度降低,转录本 4、6a 和 7 的降低最大(5 倍)。转录本 4 是导致 Mb 1501B 生长和发育发生变化的主要原因,它约占嫁接植株和块茎中总 poly(A) RNA 的 0.0003%。