Agricultural and Forestry Experiment Station, University of Alaska-Fairbanks, Fairbanks, Alaska 99775-0080.
Plant Physiol. 1986 Jun;81(2):471-7. doi: 10.1104/pp.81.2.471.
Thylakoids isolated from leaves of winter rye (Secale cereale L. cv Puma) grown at either 20 or 5 degrees C were extracted with the nonionic detergents Triton X-100 and octyl glucoside. Less total chlorophyll was extracted from 5 degrees C thylakoids by these detergents under all conditions, including pretreatment with cations. Thylakoids from either 20 or 5 degrees C leaves were solubilized in 0.7% Triton X-100 and centrifuged on sucrose gradients to purify the light harvesting complex (LHCII). Greater yields of LHCII were obtained by cation precipitation of particles derived from 20 degrees C thylakoids than from 5 degrees C thylakoids. When 20 and 5 degrees C thylakoids were phosphorylated and completely solubilized in sodium dodecyl sulfate, no differences were observed in the (32)Pi-labeling characteristics of the membrane polypeptides. However, when phosphorylated thylakoids were extracted with octyl glucoside, extraction of LHCII associated with the 5 degrees C thylakoids was markedly reduced in comparison with the extraction of LHCII from 20 degrees C membranes. Since 20 and 5 degrees C thylakoids exhibited significant differences in the Chl content and Chl a/b ratios of membrane fractions produced after solubilization with either Triton X-100 or octyl glucoside, and since few differences between the proteins of the two membranes could be observed following complete denaturation in sodium dodecyl sulfate, we conclude that the integral structure of the thylakoid membrane is affected during rye leaf development at low temperature.
从在 20℃或 5℃下生长的冬黑麦（Secale cereale L. cv Puma）叶片中分离的类囊体用非离子去污剂 Triton X-100 和辛基葡糖苷提取。在所有条件下，包括用阳离子预处理，这两种去污剂从 5℃的类囊体中提取的总叶绿素都较少。来自 20℃或 5℃叶片的类囊体在 0.7% Triton X-100 中溶解，并在蔗糖梯度上离心以纯化光捕获复合物（LHCII）。与来自 5℃类囊体的颗粒相比，来自 20℃类囊体的颗粒通过阳离子沉淀获得更高产量的 LHCII。当 20℃和 5℃的类囊体在十二烷基硫酸钠中磷酸化并完全溶解时，膜多肽的（32）Pi 标记特征没有差异。然而，当用辛基葡糖苷提取磷酸化的类囊体时，与从 20℃膜提取的 LHCII 相比，与 5℃类囊体相关的 LHCII 的提取明显减少。由于在用 Triton X-100 或辛基葡糖苷溶解后产生的膜部分中，20℃和 5℃的类囊体在 Chl 含量和 Chl a/b 比值方面表现出显著差异，并且在用十二烷基硫酸钠完全变性后几乎观察不到两个膜之间的蛋白质差异，因此我们得出结论，在低温下冬黑麦叶片发育过程中类囊体膜的整体结构受到影响。
