Hazama Kenji, Nagata Shinji, Fujimori Tamaki, Yanagisawa Shuichi, Yoneyama Tadakatsu
Department of Applied Biological Chemistry, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Physiol Plant. 2015 Jun;154(2):243-55. doi: 10.1111/ppl.12309. Epub 2014 Dec 26.
We examined the concentrations of metals (Cd, Zn, Cu, Fe and Mn) and potential metal-binding compounds [nicotianamine (NA), thiol compounds and citrate] in xylem and phloem saps from 4-week-old castor bean plants (Ricinus communis) treated with 0 (control), 0.1, 1.0, and 10 μM Cd for 3 weeks. Treatment with 0.1 and 1 μM Cd produced no visible damage, while 10 μM Cd retarded growth. Cadmium concentrations in both saps were higher than those in the culture solution at 0.1 μM, similar at 1.0 μM and lower at 10 μM. Cd at 10 μM reduced Cu and Fe concentrations in both saps. NA concentrations measured by capillary electrophoresis-mass spectrometry (MS) in xylem sap (20 μM) were higher than the Cu concentrations, and those in phloem sap (150 μM) were higher than those of Zn, Fe and Cu combined. Reduced glutathione concentrations differed in xylem and phloem saps (1-2 and 30-150 μM, respectively), but oxidized glutathione concentrations were similar. Phloem sap phytochelatin 2 concentration increased from 0.8 μM in controls to 8 μM in 10 μM Cd. Free citrate was 2-4 μM in xylem sap and 70-100 μM in phloem sap. Total bound forms of Cd in phloem and xylem saps from 1 μM Cd-treated plants were 54 and 8%, respectively. Treatment of phloem sap with proteinaseK reduced high-molecular compounds while increasing fractions of low-molecular Cd-thiol complexes. Zinc-NA, Fe-NA and Cu-NA were identified in the phloem sap fraction of control plants by electrospray ionization time-of-flight MS, and the xylem sap contained Cu-NA.
我们检测了用0(对照)、0.1、1.0和10 μM镉处理3周的4周龄蓖麻植株(Ricinus communis)木质部和韧皮部汁液中金属(镉、锌、铜、铁和锰)以及潜在的金属结合化合物[烟酰胺(NA)、硫醇化合物和柠檬酸盐]的浓度。用0.1和1 μM镉处理未产生可见损伤,而10 μM镉则抑制生长。在0.1 μM时,两种汁液中的镉浓度均高于培养液中的浓度,在1.0 μM时相似,在10 μM时较低。10 μM的镉降低了两种汁液中铜和铁的浓度。通过毛细管电泳-质谱(MS)测定,木质部汁液中NA的浓度(20 μM)高于铜的浓度,韧皮部汁液中NA的浓度(150 μM)高于锌、铁和铜浓度之和。还原型谷胱甘肽的浓度在木质部和韧皮部汁液中有所不同(分别为1 - 2和30 - 150 μM),但氧化型谷胱甘肽浓度相似。韧皮部汁液中植物螯合肽2的浓度从对照中的0.8 μM增加到10 μM镉处理时的8 μM。木质部汁液中游离柠檬酸盐为2 - 4 μM;韧皮部汁液中为70 - 100 μM。用1 μM镉处理的植株的韧皮部和木质部汁液中镉的总结合形式分别为54%和8%。用蛋白酶K处理韧皮部汁液会减少高分子化合物,同时增加低分子镉-硫醇复合物的比例。通过电喷雾电离飞行时间质谱在对照植株的韧皮部汁液组分中鉴定出锌-NA、铁-NA和铜-NA,木质部汁液中含有铜-NA。