Knowles V L, McHugh S G, Hu Z, Dennis D T, Miki B L, Plaxton W C
Department of Biology, Queen's University, Kingston, Ontario, Canada.
Plant Physiol. 1998 Jan;116(1):45-51. doi: 10.1104/pp.116.1.45.
Previously, we reported that transformation of tobacco (Nicotiana tabacum L.) with a vector containing a potato cytosolic pyruvate kinase (PKc) cDNA generated two plant lines specifically lacking leaf PKc (PKc-) as a result of co-suppression. PKc deficiency in these primary transformants did not appear to alter plant development, although root growth was not examined. Here we report a striking reduction in root growth of homozygous progeny of both PKc- lines throughout development under moderate (600 microE m-2 s-1) or low (100 microE m-2 s-1) light intensities. When both PKc- lines were cultivated under low light, shoot and flower development were also delayed and leaf indentations were apparent. Leaf PK activity in the transformants was significantly decreased at all time points examined, whereas root activities were unaffected. Polypeptides corresponding to PKc were undetectable on immunoblots of PKc- leaf extracts, except in 6-week-old low-light-grown PKc- plants, in which leaf PKc expression appeared to be greatly reduced. The metabolic implications of the kinetic characteristics of partially purified PKc from wild-type tobacco leaves are discussed. Overall, the results suggest that leaf PKc deficiency leads to a perturbation in source-sink relationships.
此前,我们报道过,用含有马铃薯胞质丙酮酸激酶(PKc)cDNA的载体转化烟草(Nicotiana tabacum L.),由于共抑制作用产生了两个特异性缺乏叶片PKc的植株系(PKc-)。这些初级转化体中的PKc缺陷似乎并未改变植株发育,不过未检测根系生长情况。在此我们报道,在中等(600微爱因斯坦·米-2·秒-1)或低(100微爱因斯坦·米-2·秒-1)光照强度下,两个PKc-系的纯合子后代在整个发育过程中根系生长显著降低。当两个PKc-系在低光照条件下培养时,地上部和花的发育也延迟,叶片出现缺刻。在所有检测的时间点,转化体中的叶片PK活性显著降低,而根系活性未受影响。除了6周龄低光照生长的PKc-植株中叶片PKc表达似乎大幅降低外,在PKc-叶片提取物的免疫印迹上未检测到与PKc对应的多肽。讨论了从野生型烟草叶片中部分纯化的PKc的动力学特征的代谢意义。总体而言,结果表明叶片PKc缺陷导致源库关系紊乱。