本研究从马铃薯(Solanum tuberosum cv.Chieftain)茎的愈伤组织中分离得到绿色、白色和红色3种愈伤组织,并利用鲜重法和分光光度法分别测量愈伤组织的生长量、叶绿素和花色苷的含量;通过半定量RT—PCR法分析4个花色苷生物合成相关基因的表达差异。结果表明:绿色愈伤组织生长最快,叶绿素含量最高,红色愈伤组织生长最慢,叶绿素含量最低;绿色和白色愈伤组织中几乎不含花色苷,红色愈伤组织中花色苷含量最高,达2.1OD513/mgFw;对CHS、F3H、DFR和F3’5’H四个花色苷生物合成相关基因表达分析发现,绿色和白色愈伤组织不合成花色苷可能与DFR基因不表达有关。本实验结果为进一步阐明花色昔生物合成机理和花色苷色素的生产应用提供一定的理论依据。
The green callus, white callus and red callus were isolated from the stem callus of potatoes (Solarium tuberosum cv. Chieftain). Gallus growth, chlorophyll and anthocyanin content in callus cultures were measured by using fresh weight and spectrophotometric method. The differential expression of four anthocyanin biosynthetic genes was investigated by semi-quantitative RT-PCR. The results indicated that the growth of green callus were the fastest, the contents of chlorophyll were the most, while that the growth of red callus were the slowest. The contents of chlorophyll were the lowest. The contents of chlorophyll and anthocyanins were significantly different in three different calluses. The contents ofanthocyanin in green and white callus were very low, while high amount of anthocyanin was found in red callus. The content of anthocyanins reached 2.1OD513/mg FW in the red callus. The expression analysis of CHS, F3H, F3'5'H and DFR genes in three different callus showed that no DFR expression were detected in the green and white callus, this may be led to accumulate no anthocyanin. The results of this experiment will provide a theoretical basis for further clarity the mechanism ofanthocyanin biosynthesis and application.