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大豆果实去除诱导叶片提取物中形成核酮糖-1,5-二磷酸羧化酶/加氧酶的不溶性形式*。

Fruit removal in soybean induces the formation of an insoluble form of ribulose-1,5-bisphosphate carboxylase/oxygenase in leaf extracts*.

机构信息

U.S. Department of Agriculture, Agricultural Research Service, USA.

出版信息

Planta. 1991 Jan;183(2):300-6. doi: 10.1007/BF00197802.

Abstract

In some soybean (Glycine max (L.) Merr.) cultivars, fruit removal does not delay the apparent loss of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) activity and abundance or the decline in photosynthesis. Analysis of leaf extracts from defruited plants indicated a time-dependent increase in both Rubisco activity and abundance in a 30000 · g pellet fraction in cultivars which had been reported to lose all Rubisco protein from the supernatant fraction. Attempts to solubilize the pelleted Rubisco by increasing the buffer volume/tissue ratio or by adding alkylphenoxypolyethoxyethanol (Triton X-100), ethylenediaminetetraacetic acid (EDTA), or NaCl were unsuccessful. However, treatment of the pellets with denaturants such as 8 M urea or 5% (w/v) sodium dodecyl sulfate (SDS) did release Rubisco from the pellet. Redistribution of protein to the pellet fraction appeared to be specific for Rubisco since the amount of ribulose-5-phosphate kinase (EC 2.7.1.19) found in the pellet fraction of leaf extracts of control and defruited plants was small and constant over time. The loss of soluble Rubisco, and the concomitant increase in insoluble Rubisco, in response to fruit removal varied with genotype and was reproducible in both field and greenhouse environments. In addition, the effect was influenced by node position and light; lower and-or shaded leaves exhibited less Rubisco in the pellet fraction than leaves from the top of the plant that was fully exposed to sunlight. When isolated by sucrose-density-gradient centrifugation, the insoluble Rubisco was found to co-purify with a 30-kDa (kilodalton) polypeptide. These results indicate that alteration of the source/sink ratio by removing fruits results in the formation of an insoluble form of Rubisco in leaf extracts of soybean. Whether or not Rubisco exists as an insoluble complex with the 30-kDa polypeptide in intact leaves of defruited plants remains to be determined.

摘要

在一些大豆(Glycine max (L.) Merr.)品种中,去除果实不会延迟核酮糖-1,5-二磷酸羧化酶/加氧酶(Rubisco,EC 4.1.1.39)活性和丰度的明显损失,也不会导致光合作用下降。对去果植株叶片提取物的分析表明,在已报道上清液中失去所有Rubisco 蛋白的品种中,Rubisco 活性和丰度在 30000g 沉淀部分随时间呈依赖性增加。通过增加缓冲液/组织比或添加烷基酚聚氧乙烯醚(Triton X-100)、乙二胺四乙酸(EDTA)或 NaCl 来尝试溶解沉淀的 Rubisco 均未成功。然而,用变性剂如 8 M 尿素或 5%(w/v)十二烷基硫酸钠(SDS)处理沉淀可以将 Rubisco 从沉淀中释放出来。蛋白质重新分配到沉淀部分似乎是 Rubisco 特有的,因为在对照和去果植株叶片提取物的沉淀部分中发现的核酮糖-5-磷酸激酶(EC 2.7.1.19)的量很小且随时间保持不变。可溶性 Rubisco 的损失以及对果实去除的不溶性 Rubisco 的相应增加因基因型而异,并且在田间和温室环境中均具有可重复性。此外,该效应受节位和光照的影响;与充分暴露在阳光下的植株顶部的叶片相比,较低和/或遮荫的叶片在沉淀部分的 Rubisco 含量较少。当通过蔗糖密度梯度离心分离时,发现不溶性 Rubisco 与 30kDa(千道尔顿)多肽共同纯化。这些结果表明,通过去除果实来改变源/库比会导致大豆叶片提取物中 Rubisco 形成不溶性形式。在去果植株的完整叶片中,Rubisco 是否以不溶性复合物的形式存在与 30kDa 多肽仍有待确定。

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