Chloroplast Cytochrome b6/f and ATP Synthase Complexes in Tobacco: Transformation With Antisense RNA Against Nuclear-Encoded Transcripts for the Rieske FeS and ATPδ Polypeptides
Chloroplast Cytochrome b6/f and ATP Synthase Complexes in Tobacco: Transformation With Antisense RNA Against Nuclear-Encoded Transcripts for the Rieske FeS and ATPδ Polypeptides
复制标题
烟草中叶绿体细胞色素 b6/f 和 ATP 合酶复合物：用反义 RNA 转化 Rieske FeS 和 ATPδ 多肽的核编码转录本
DOI：
10.1071/pp9950285
复制
发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Badger
中科院分区：
文献类型：
--
作者：
G. Price;Jian;S. Caemmerer;J. R. Evans;W. Chow;Jan M. Anderson;V. Hurry;Badger
利用反义RNA技术已经产生了叶绿体细胞色素b6/f和ATP合酶复合物的量和活性都降低的烟草植物。从分别编码Rieske FeS蛋白和b6/f和ATP合酶复合物的δ亚基的烟草cDNA克隆产生反义构建体。使用脉冲调制荧光测量选择具有改变的活性的转化体。b6/f转化体表现出高水平的稳态荧光和降低水平的光化学和非光化学淬灭。与此形成鲜明对比的是，ATP合酶转化体表现出低水平的稳态荧光和大大增加的非光化学猝灭水平。对于两种构建体分离具有一系列抑制的转化体，在某些情况下光合作用降低至野生型值的10%以下。受影响最严重的转化体表现出极慢的生长，在某些情况下，它们不能生长和产生种子。从R1种子从几个细胞色素b6/f转化体的后代进行了分析，并显示分离的表型范围从中间到严重的镇压。中度和重度表型植物显示Rieske FeS mRNA减少超过90%，而FeS多肽减少至野生型的60%和86%。在空气中的光合作用和Rieske FeS多肽含量在反义植物中有很强的相关性，这表明细胞色素b6/f复合物是在这些条件下光合速率的主要决定因素。FeS反义植物的光抑制研究表明，在最严重的植物中叶黄素循环的活性降低，与transthylakoid pH梯度的降低和非光化学淬灭的降低一致。对ATPδ反义植物的初步研究表明，它们也具有降低的mRNA和ATPδ多肽水平。
Tobacco plants with reduced amounts and activities of both the chloroplast cytochrome b6/f and ATP synthase complexes have been produced using antisense RNA techniques. Antisense constructs were generated from tobacco cDNA clones coding for the Rieske FeS protein and the δ subunit of the b6/f and ATP synthase complexes respectively. Transformants with altered activities were selected using pulse-modulated fluorescence measurements. The b6/ftransformants showed high levels of steady-state fluorescence and reduced levels of both photochemical and non-photochemical quenching. In striking contrast, the ATP synthase transformants showed low levels of steady-state fluorescence and greatly increased levels of non-photochemical quenching. Transformants with a range of suppression were isolated for both constructs, in some cases with photosynthesis reduced to less than 10% of wild-type values. The most severely affected transformants showed extremely slow growth and in some cases they were unable to grow and produce seed. Progeny from the R1 seed from several cytochrome b6/f transformants have been analysed and show segregation of phenotypes ranging from intermediate to severe in repression. Intermediate and severe phenotype plants showed a reduction in Rieske FeS mRNA of more than 90% while FeS polypeptide was reduced to 60 and 86% of wild type. There was a strong correlation between photosynthesis at air and Rieske FeS polypeptide content in the antisense plants suggesting that the cytochrome b6/f complex was a major determinant of photosynthetic rate under these conditions. Photoinhibition studies of FeS antisense plants showed that there was a reduced activity of the xanthophyll cycle in the most severe plants, consistent with a reduction in the transthylakoid pH gradient and a lowered non-photochemical quenching. Preliminary studies of the ATPδ antisense plants showed that they also had reduced levels of mRNA and ATPδ polypeptide.