Chiaiese Pasquale, Ohkama-Ohtsu Naoko, Molvig Lisa, Godfree Robert, Dove Hugh, Hocart Charles, Fujiwara Toru, Higgins T J V, Tabe Linda M
CSIRO Plant Industry, GPO Box 1600, Canberra ACT 2601, Australia.
J Exp Bot. 2004 Aug;55(404):1889-901. doi: 10.1093/jxb/erh198. Epub 2004 Jul 2.
In order to increase the concentration of the nutritionally essential sulphur amino acids in seed protein, a transgene encoding a methionine- and cysteine-rich protein, sunflower seed albumin (SSA), was transferred to chickpeas (Cicer arietinum L). Transgenic seeds that accumulated SSA contained more methionine and less oxidized sulphur than the controls, suggesting that additional demand for sulphur amino acids from the expression of the transgene stimulated sulphur assimilation. In addition, the activity of trypsin inhibitors, a known family of endogenous, sulphur-rich chickpea seed proteins, was diminished in transgenic, SSA-containing seeds compared with the non-transgenic controls. Together, these results indicate that the reduced sulphur sequestered into SSA was supplied partly by additional sulphur assimilation in the developing transgenic seeds, and partly by some diversion of sulphur amino acids from endogenous seed proteins. Growth of chickpeas on nutrient with a high sulphur-to-nitrogen ratio increased the total seed sulphur content and the accumulation of sulphur amino acids in the seeds, and partly mitigated the effect of SSA accumulation on the trypsin inhibitor amount. The results suggest that free methionine and O-acetylserine (OAS) acted as signals that modulated chickpea seed protein composition in response to the variation in sulphur demand, as well as in response to variation in the nitrogen and sulphur status of the plant.
为了提高种子蛋白中营养必需的含硫氨基酸的浓度,将编码富含蛋氨酸和半胱氨酸的蛋白质——向日葵种子清蛋白(SSA)的转基因导入鹰嘴豆(Cicer arietinum L)。积累SSA的转基因种子比对照含有更多的蛋氨酸和更少的氧化态硫,这表明转基因表达对含硫氨基酸的额外需求刺激了硫同化作用。此外,与非转基因对照相比,转基因的、含有SSA的种子中胰蛋白酶抑制剂(一种已知的内源性、富含硫的鹰嘴豆种子蛋白家族)的活性降低。这些结果共同表明,转入SSA中减少的硫部分由发育中的转基因种子中额外的硫同化作用提供,部分由内源性种子蛋白中硫氨基酸的一些转移提供。在高硫氮比的养分上种植鹰嘴豆增加了种子的总硫含量和种子中含硫氨基酸的积累,并部分减轻了SSA积累对胰蛋白酶抑制剂含量的影响。结果表明,游离蛋氨酸和O-乙酰丝氨酸(OAS)作为信号,响应硫需求的变化以及植物氮和硫状态的变化,调节鹰嘴豆种子蛋白的组成。