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首页> 外文期刊>Vitis >CO_2/O_2 specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase in grapevines (Vitis vinifera L.): First in vitro determination and comparison to in vivo estimations
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CO_2/O_2 specificity factor of ribulose-1,5-bisphosphate carboxylase/oxygenase in grapevines (Vitis vinifera L.): First in vitro determination and comparison to in vivo estimations

机译：葡萄（葡萄）的核糖-1,5-双磷酸羧化酶/加氧酶的CO_2 / O_2特异性因子：首次体外测定并与体内估计值进行比较

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摘要

The specificity factor (S) of ribulose-l,5-bisphosphate carboxylase/oxygenase (Rubisco) defines the relative rates of carboxylation and oxygenation of Ribulose-1,5-bisphos-phate (RuBP) catalysed by the enzyme. The determination of S for Rubisco purified from the Vitis vinifera L. cvs Tempranillo and Manto Negro is described here for the first time. Rubisco extraction was made in Bicine buffer with the inclusion of polyethylene glycol (PEG), β-mercaptoethanol, diethyldithio-carbamic acid (DIECA) and several protease inhibitors. Furthermore, in the same cultivars, the apparent in vivo specificity factor for Rubisco (S*), was obtained from gas exchange and chlorophyll fluorescence measurements. For both cultivars the values of S were close to 100 at 25℃. However, in mature leaves, S* was about 67 for Manto Negro and 55 or 77 for Tempranillo, depending on leaf age. Leaves of plants under drought showed even lower values. These discrepancies between S and S* are ascribed to equating CO_2 in the sub-stomatal cavity with CO_2 at the Rubisco catalytic site in the chloroplast. However, S* values from young developing leaves were very close to S for both cultivars. It is concluded that estimations of S* based on gas exchange and chlorophyll fluorescence data are reliable only in thin, young and non-stressed leaves.
机译：核糖-1，5-双磷酸羧化酶/加氧酶（Rubisco）的特异性因子（S）定义了由该酶催化的-1,5-双磷酸膦酸酯（RuBP）的羧化和氧合的相对速率。在此首次描述了从葡萄属葡萄球菌Tempranillo和Manto Negro纯化的Rubisco中S的测定。 Rubisco提取物在Bicine缓冲液中提取，其中包含聚乙二醇（PEG），β-巯基乙醇，二乙基二硫代氨基甲酸（DIECA）和几种蛋白酶抑制剂。此外，在相同品种中，通过气体交换和叶绿素荧光测量获得了对Rubisco（S *）的表观体内特异性因子。两种品种在25℃下的S值均接近100。但是，在成熟叶片中，Manto Negro的S *约为67，坦普拉尼约的S *约为55或77，具体取决于叶片的年龄。干旱下的植物叶片显示出更低的值。 S和S *之间的这些差异归因于使下气孔腔内的CO_2等于叶绿体中Rubisco催化位点处的CO_2。但是，两个品种的幼叶的S *值都非常接近S。结论是，基于气体交换和叶绿素荧光数据的S *估算仅在薄，年轻和非胁迫叶片中是可靠的。

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